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Sample records for enhance bead-based bioassays

  1. Use of a Novel Fluidics Microbead Trap/Flow-cell Enhances Speed and Sensitivity of Bead-Based Bioassays

    SciTech Connect

    Ozanich, Rich M.; Antolick, Kathryn C.; Bruckner-Lea, Cindy J.; Dockendorff, Brian P.; Easterday, Ashton N.; Edberg, Heather C.; Grate, Jay W.; Iyer, Sujata; Johnson, Laura H.; Straub, Tim M.; Valdez, Catherine O.; Warner, Marvin G.

    2007-09-15

    Automated devices and methods for biological sample preparation often utilize surface functionalized microbeads (superparamagnetic or non-magnetic) to allow capture, purification and pre-concentration of trace amounts of proteins, cells, or nucleic acids (DNA/RNA) from complex samples. We have developed unique methods and hardware for trapping either magnetic or non-magnetic functionalized beads that allow samples and reagents to be efficiently perfused over a micro-column of beads. This approach yields enhanced mass transport and up to 5-fold improvements in assay sensitivity or speed, dramatically improving assay capability relative to assays conducted in more traditional “batch modes” (i.e., in tubes or microplate wells). Summary results are given that highlight the analytical performance improvements obtained for automated microbead processing systems utilizing novel microbead trap/flow-cells for various applications, including: 1) simultaneous capture of multiple cytokines using an antibody-coupled polystyrene bead assay with subsequent flow cytometry detection; 2) capture of nucleic acids using oligonucleotide coupled polystyrene beads with flow cytometry detection; and 3) capture of Escherichia coli 0157:H7 (E. coli) from 50 mL sample volumes using antibody-coupled superparamagnetic microbeads with subsequent culturing to assess capture efficiency.

  2. An optoelectrokinetic technique for programmable particle manipulation and bead-based biosignal enhancement.

    PubMed

    Wang, Kuan-Chih; Kumar, Aloke; Williams, Stuart J; Green, Nicolas G; Kim, Kyung Chun; Chuang, Han-Sheng

    2014-10-21

    Technologies that can enable concentration of low-abundance biomarkers are essential for early diagnosis of diseases. In this study, an optoelectrokinetic technique, termed Rapid Electrokinetic Patterning (REP), was used to enable dynamic particle manipulation in bead-based bioassays. Various manipulation capabilities, such as micro/nanoparticle aggregation, translation, sorting and patterning, were developed. The technique allows for versatile multi-parameter (voltage, light intensity and frequency) based modulation and dynamically addressable manipulation with simple device fabrication. Signal enhancement of a bead-based bioassay was demonstrated using dilute biotin-fluorescein isothiocyanate (FITC) solutions mixed with streptavidin-conjugated particles and rapidly concentrated with the technique. As compared with a conventional ELISA reader, the REP-enabled detection achieved a minimal readout of 3.87 nM, which was a 100-fold improvement in sensitivity. The multi-functional platform provides an effective measure to enhance detection levels in more bead-based bioassays. PMID:25109364

  3. Enhancement of performance in porous bead-based microchip sensors: Effects of chip geometry on bio-agent capture

    PubMed Central

    Kulla, Eliona; Chou, Jie; Simmons, Glennon; Wong, Jorge; McRae, Michael P.; Patel, Rushi; Floriano, Pierre N.; Christodoulides, Nicolaos; Leach, Robin J.; Thompson, Ian M.; McDevitt, John T.

    2015-01-01

    Measuring low concentrations of clinically-important biomarkers using porous bead-based lab-on-a-chip (LOC) platforms is critical for the successful implementation of point-of-care (POC) devices. One way to meet this objective is to optimize the geometry of the bead holder, referred to here as a micro-container. In this work, two geometric micro-containers were explored, the inverted pyramid frustum (PF) and the inverted clipped pyramid frustum (CPF). Finite element models of this bead array assay system were developed to optimize the micro-container and bead geometries for increased pressure, to increase analyte capture in porous bead-based fluorescence immunoassays. Custom micro-milled micro-container structures containing an inverted CPF geometry resulted in a 28% reduction in flow-through regions from traditional anisotropically-etched pyramidal geometry derived from Si-111 termination layers. This novel “reduced flow-through” design resulted in a 33% increase in analyte penetration into the bead and twofold increase in fluorescence signal intensity as demonstrated with C-Reactive Protein (CRP) antigen, an important biomarker of inflammation. A consequent twofold decrease in the limit of detection (LOD) and the limit of quantification (LOQ) of a proof-of-concept assay for the free isoform of Prostate-Specific Antigen (free PSA), an important biomarker for prostate cancer detection, is also presented. Furthermore, a 53% decrease in the bead diameter is shown to result in a 160% increase in pressure and 2.5-fold increase in signal, as estimated by COMSOL models and confirmed experimentally by epi-fluorescence microscopy. Such optimizations of the bead micro-container and bead geometries have the potential to significantly reduce the LODs and reagent costs for spatially programmed bead-based assay systems of this type. PMID:26097696

  4. Sensitivity Enhancement of Bead-based Electrochemical Impedance Spectroscopy (BEIS) biosensor by electric field-focusing in microwells.

    PubMed

    Shin, Kyeong-Sik; Ji, Jae Hoon; Hwang, Kyo Seon; Jun, Seong Chan; Kang, Ji Yoon

    2016-11-15

    This paper reports a novel electrochemical impedance spectroscopy (EIS) biosensors that uses magnetic beads trapped in a microwell array to improve the sensitivity of conventional bead-based EIS (BEIS) biosensors. Unloading the previously measured beads by removing the magnetic bar enables the BEIS sensor to be used repeatedly by reloading it with new beads. Despite its recyclability, the sensitivity of conventional BEIS biosensors is so low that it has not attracted much attentions from the biosensor industry. We significantly improved the sensitivity of the BEIS system by introducing of a microwell array that contains two electrodes (a working electrode and a counter electrode) to concentrate the electric field on the surfaces of the beads. We confirmed that the performance of the BEIS sensor in a microwell array using an immunoassay of prostate specific antigen (PSA) in PBS buffer and human plasma. The experimental results showed that a low concentration of PSA (a few tens or hundreds of fg/mL) were detectable as a ratio of the changes in the impedance of the PBS buffer or in human plasma. Therefore, our BEIS sensor with a microwell array could be a promising platform for low cost, high-performance biosensors for applications that require high sensitivity and recyclability. PMID:27152445

  5. Encapsulation of Adenovirus Serotype 5 in Anionic Lecithin Liposomes using a Bead-Based Immunoprecipitation Technique Enhances Transfection Efficiency

    PubMed Central

    Mendez, N.; Herrera, V.; Zhang, L.; Hedjran, F.; Feuer, R.; Blair, S.; Trogler, W.; Reid, T.

    2014-01-01

    Oncolytic viruses (OVs) constitute a promising class of cancer therapeutics which exploit validated genetic pathways known to be deregulated in many cancers. To overcome an immune response and to enhance its potential use to treat primary and metastatic tumors, a method for liposomal encapsulation of adenovirus has been developed. The encapsulation of adenovirus in non-toxic anionic lecithin-cholesterol-PEG liposomes ranging from 140–180nm in diameter have been prepared by self-assembly around the viral capsid. The encapsulated viruses retain their ability to infect cancer cells. Furthermore, an immunoprecipitation (IP) technique has shown to be a fast and effective method to extract non-encapsulated viruses and homogenize the liposomes remaining in solution. 78% of adenovirus plaque forming units were encapsulated and retained infectivity after IP processing. Additionally, encapsulated viruses have shown enhanced transfection efficiency up to 4× higher compared to non-encapsulated Ads. Extracting non-encapsulated viruses from solution may prevent an adverse in vivo immune response and may enhance treatment for multiple administrations. PMID:25154663

  6. Microfluidic, Bead-Based Assay: Theory and Experiments

    PubMed Central

    Thompson, Jason A.; Bau, Haim H.

    2009-01-01

    Microbeads are frequently used as a solid support for biomolecules such as proteins and nucleic acids in heterogeneous microfluidic assays. However, relatively few studies investigate the binding kinetics on modified bead surfaces in a microfluidics context. In this study, a customized hot embossing technique is used to stamp microwells in a thin plastic substrate where streptavidin-coated agarose beads are selectively placed and subsequently immobilized within a conduit. Biotinylated quantum dots are used as a label to monitor target analyte binding to the bead's surface. Three-dimensional finite element simulations are carried out to model the binding kinetics on the bead's surface. The model accounts for surface exclusion effects resulting from a single quantum dot occluding multiple receptor sites. The theoretical predictions are compared and favorably agree with experimental observations. The theoretical simulations provide a useful tool to predict how varying parameters affect microbead reaction kinetics and sensor performance. This study enhances our understanding of bead-based microfluidic assays and provides a design tool for developers of point-of-care, lab-on-chip devices for medical diagnosis, food and water quality inspection, and environmental monitoring. PMID:19766545

  7. Enhanced fluorescence of proteins and label-free bioassays using aluminum nanostructures.

    PubMed

    Ray, Krishanu; Szmacinski, Henryk; Lakowicz, Joseph R

    2009-08-01

    We report the enhanced intrinsic fluorescence from several proteins in proximity to aluminum nanostructured surfaces. Intrinsic fluorescence in proteins is dominated by the tryptophan residues. Intensities and lifetimes of several proteins with different numbers of tryptophan residues assembled on the surfaces of quartz or aluminum nanostructured films were measured. Immobilized protein molecules on the surface of an aluminum nanostructured film resulted in a significant fluorescence intensity enhancement (up to 14-fold) and lifetime decrease (up to 6-fold) compared to the quartz substrates. These large spectroscopic changes allow design of label-free bioassays where detection of binding interactions between proteins can be observed in the presence of a bulk sample solution. Binding of streptavidin to the biotinylated aluminum surface was demonstrated in the presence of 100 microg/mL bovine serum albumin in the sample solution by measurements of tryptophan intensity and lifetime changes. PMID:19594133

  8. High performance wash-free magnetic bioassays through microfluidically enhanced particle specificity.

    PubMed

    Bechstein, Daniel J B; Lee, Jung-Rok; Ooi, Chin Chun; Gani, Adi W; Kim, Kyunglok; Wilson, Robert J; Wang, Shan X

    2015-01-01

    Magnetic biosensors have emerged as a sensitive and versatile platform for high performance medical diagnostics. These magnetic biosensors require well-tailored magnetic particles as detection probes, which need to give rise to a large and specific biological signal while showing very low nonspecific binding. This is especially important in wash-free bioassay protocols, which do not require removal of particles before measurement, often a necessity in point of care diagnostics. Here we show that magnetic interactions between magnetic particles and magnetized sensors dramatically impact particle transport and magnetic adhesion to the sensor surfaces. We investigate the dynamics of magnetic particles' biomolecular binding and magnetic adhesion to the sensor surface using microfluidic experiments. We elucidate how flow forces can inhibit magnetic adhesion, greatly diminishing or even eliminating nonspecific signals in wash-free magnetic bioassays, and enhancing signal to noise ratios by several orders of magnitude. Our method is useful for selecting and optimizing magnetic particles for a wide range of magnetic sensor platforms. PMID:26123868

  9. High performance wash-free magnetic bioassays through microfluidically enhanced particle specificity

    PubMed Central

    Bechstein, Daniel J.B.; Lee, Jung-Rok; Ooi, Chin Chun; Gani, Adi W.; Kim, Kyunglok; Wilson, Robert J.; Wang, Shan X.

    2015-01-01

    Magnetic biosensors have emerged as a sensitive and versatile platform for high performance medical diagnostics. These magnetic biosensors require well-tailored magnetic particles as detection probes, which need to give rise to a large and specific biological signal while showing very low nonspecific binding. This is especially important in wash-free bioassay protocols, which do not require removal of particles before measurement, often a necessity in point of care diagnostics. Here we show that magnetic interactions between magnetic particles and magnetized sensors dramatically impact particle transport and magnetic adhesion to the sensor surfaces. We investigate the dynamics of magnetic particles’ biomolecular binding and magnetic adhesion to the sensor surface using microfluidic experiments. We elucidate how flow forces can inhibit magnetic adhesion, greatly diminishing or even eliminating nonspecific signals in wash-free magnetic bioassays, and enhancing signal to noise ratios by several orders of magnitude. Our method is useful for selecting and optimizing magnetic particles for a wide range of magnetic sensor platforms. PMID:26123868

  10. High performance wash-free magnetic bioassays through microfluidically enhanced particle specificity

    NASA Astrophysics Data System (ADS)

    Bechstein, Daniel J. B.; Lee, Jung-Rok; Ooi, Chin Chun; Gani, Adi W.; Kim, Kyunglok; Wilson, Robert J.; Wang, Shan X.

    2015-06-01

    Magnetic biosensors have emerged as a sensitive and versatile platform for high performance medical diagnostics. These magnetic biosensors require well-tailored magnetic particles as detection probes, which need to give rise to a large and specific biological signal while showing very low nonspecific binding. This is especially important in wash-free bioassay protocols, which do not require removal of particles before measurement, often a necessity in point of care diagnostics. Here we show that magnetic interactions between magnetic particles and magnetized sensors dramatically impact particle transport and magnetic adhesion to the sensor surfaces. We investigate the dynamics of magnetic particles’ biomolecular binding and magnetic adhesion to the sensor surface using microfluidic experiments. We elucidate how flow forces can inhibit magnetic adhesion, greatly diminishing or even eliminating nonspecific signals in wash-free magnetic bioassays, and enhancing signal to noise ratios by several orders of magnitude. Our method is useful for selecting and optimizing magnetic particles for a wide range of magnetic sensor platforms.

  11. Properties of coatings on RFID p-Chips that support plasmonic fluorescence enhancement in bioassays

    PubMed Central

    Rich, Ryan; Li, Ji; Fudala, Rafal; Gryczynski, Zygmunt; Gryczynski, Ignacy; Mandecki, Wlodek

    2012-01-01

    Microtransponders (RFID p-Chips) derivatized with silver island film (SIF) have previously seen success as a platform for the quantification of low-abundance biomolecules in nucleic acid-based assays and immunoassays. In this study, we further characterized the morphology of the SIF as well as the polymer matrix enveloping it by scanning electron microscopy (SEM). The polymer was a two-layer silane-based matrix engulfing the p-Chip and SIF. Through a series of SEM and confocal fluorescence microscopy experiments we found the depth of the polymer matrix to be 1–2 µm. The radiative effects of the SIF/polymer layer were assessed by fluorescence lifetime imaging (FLIM) of p-Chips coated with the polymer to which a fluorophore (Alexa Fluor 555) was conjugated. FLIM images showed an 8.7-fold increase in fluorescence intensity and an increased rate of radiative decay, the latter of which is associated with improved photostability and both of which are linked to plasmonic enhancement by the SIF. Plasmonic enhancement was found to extend uniformly across the p-Chip and, interestingly, to a depth of about 1.2 µm. The substantial depth of enhancement suggests that the SIF/polymer layer constitutes a three-dimensional matrix that is accessible to solvent and small molecules such as fluorescent dyes. Finally, we confirmed that no surface-enhanced Raman scattering (SERS) is seen from the SIF/polymer combination. The analysis provides a possible mechanism by which the SIF/polymer-coated p-Chips allow a highly sensitive immunoassay and, as a result, leads to an improved bioassay platform. PMID:22960796

  12. Chip-Scale Bioassays Based on Surface-Enhanced Raman Scattering: Fundamentals and Applications

    SciTech Connect

    Hye-Young Park

    2005-12-17

    This work explores the development and application of chip-scale bioassays based on surface-enhanced Raman scattering (SERS) for high throughput and high sensitivity analysis of biomolecules. The size effect of gold nanoparticles on the intensity of SERS is first presented. A sandwich immunoassay was performed using Raman-labeled immunogold nanoparticles with various sizes. The SERS responses were correlated to particle densities, which were obtained by atomic force microscopy (AFM). The response of individual particles was also investigated using Raman-microscope and an array of gold islands on a silicon substrate. The location and the size of individual particles were mapped using AFM. The next study describes a low-level detection of Escherichia coli 0157:H7 and simulants of biological warfare agents in a sandwich immunoassay format using SERS labels, which have been termed Extrinsic Raman labels (ERLs). A new ERL scheme based on a mixed monolayer is also introduced. The mixed monolayer ERLs were created by covering the gold nanoparticles with a mixture of two thiolates, one thiolate for covalently binding antibody to the particle and the other thiolate for producing a strong Raman signal. An assay platform based on mixed self-assembled monolayers (SAMs) on gold is then presented. The mixed SAMs were prepared from dithiobis(succinimidyl undecanoate) (DSU) to covalently bind antibodies on gold substrate and oligo(ethylene glycol)-terminated thiol to prevent nonspecific adsorption of antibodies. After the mixed SAMs surfaces, formed from various mole fraction of DSU were incubated with antibodies, AFM was used to image individual antibodies on the surface. The final study presents a collaborative work on the single molecule adsorption of YOYO-I labeled {lambda}-DNA at compositionally patterned SAMs using total internal reflection fluorescence microscopy. The role of solution pH, {lambda}-DNA concentration, and domain size was investigated. This work also revealed

  13. Bead-based microfluidic immunoassay for diagnosis of Johne's disease

    SciTech Connect

    Wadhwa, Ashutosh; Foote, Robert; Shaw, Robert W; Eda, Shigetoshi

    2012-01-01

    Microfluidics technology offers a platform for development of point-of-care diagnostic devices for various infectious diseases. In this study, we examined whether serodiagnosis of Johne s disease (JD) can be conducted in a bead-based microfluidic assay system. Magnetic micro-beads were coated with antigens of the causative agent of JD, Mycobacterium avium subsp. paratuberculosis. The antigen-coated beads were incubated with serum samples of JD-positive or negative serum samples and then with a fluorescently-labeled secondary antibody (SAB). To confirm binding of serum antibodies to the antigen, the beads were subjected to flow cytometric analysis. Different conditions (dilutions of serum and SAB, types of SAB, and types of magnetic beads) were optimized for a great degree of differentiation between the JD-negative and JD-positive samples. Using the optimized conditions, we tested a well-classified set of 155 serum samples from JD negative and JD-positive cattle by using the bead-based flow cytometric assay. Of 105 JD-positive samples, 63 samples (60%) showed higher antibody binding levels than a cut-off value determined by using antibody binding levels of JD-negative samples. In contrast, only 43-49 JD-positive samples showed higher antibody binding levels than the cut-off value when the samples were tested by commercially-available immunoassays. Microfluidic assays were performed by magnetically immobilizing a number of beads within a microchannel of a glass microchip and detecting antibody on the collected beads by laser-induced fluorescence. Antigen-coated magnetic beads treated with bovine serum sample and fluorescently-labeled SAB were loaded into a microchannel to measure the fluorescence (reflecting level of antibody binding) on the beads in the microfluidic system. When the results of five bovine serum samples obtained with the system were compared to those obtained with the flow cytometer, a high level of correlation (linear regression, r2 = 0.994) was

  14. A dynamic bead-based microarray for parallel DNA detection

    NASA Astrophysics Data System (ADS)

    Sochol, R. D.; Casavant, B. P.; Dueck, M. E.; Lee, L. P.; Lin, L.

    2011-05-01

    A microfluidic system has been designed and constructed by means of micromachining processes to integrate both microfluidic mixing of mobile microbeads and hydrodynamic microbead arraying capabilities on a single chip to simultaneously detect multiple bio-molecules. The prototype system has four parallel reaction chambers, which include microchannels of 18 × 50 µm2 cross-sectional area and a microfluidic mixing section of 22 cm length. Parallel detection of multiple DNA oligonucleotide sequences was achieved via molecular beacon probes immobilized on polystyrene microbeads of 16 µm diameter. Experimental results show quantitative detection of three distinct DNA oligonucleotide sequences from the Hepatitis C viral (HCV) genome with single base-pair mismatch specificity. Our dynamic bead-based microarray offers an effective microfluidic platform to increase parallelization of reactions and improve microbead handling for various biological applications, including bio-molecule detection, medical diagnostics and drug screening.

  15. Rapid Bead-Based Antimicrobial Susceptibility Testing by Optical Diffusometry

    PubMed Central

    Chung, Chih-Yao; Wang, Jhih-Cheng; Chuang, Han-Sheng

    2016-01-01

    This study combined optical diffusometry and bead-based immunoassays to develop a novel technique for quantifying the growth of specific microorganisms and achieving rapid AST. Diffusivity rises when live bacteria attach to particles, resulting in additional energy from motile microorganisms. However, when UV-sterilized (dead) bacteria attach to particles, diffusivity declines. The experimental data are consistent with the theoretical model predicted according to the equivalent volume diameter. Using this diffusometric platform, the susceptibility of Pseudomonas aeruginosa to the antibiotic gentamicin was tested. The result suggests that the proliferation of bacteria is effectively controlled by gentamicin. This study demonstrated a sensitive (one bacterium on single particles) and time-saving (within 2 h) platform with a small sample volume (~0.5 μL) and a low initial bacteria count (50 CFU per droplet ~ 105 CFU/mL) for quantifying the growth of microorganisms depending on Brownian motion. The technique can be applied further to other bacterial strains and increase the success of treatments against infectious diseases in the near future. PMID:26863001

  16. Rapid Bead-Based Antimicrobial Susceptibility Testing by Optical Diffusometry.

    PubMed

    Chung, Chih-Yao; Wang, Jhih-Cheng; Chuang, Han-Sheng

    2016-01-01

    This study combined optical diffusometry and bead-based immunoassays to develop a novel technique for quantifying the growth of specific microorganisms and achieving rapid AST. Diffusivity rises when live bacteria attach to particles, resulting in additional energy from motile microorganisms. However, when UV-sterilized (dead) bacteria attach to particles, diffusivity declines. The experimental data are consistent with the theoretical model predicted according to the equivalent volume diameter. Using this diffusometric platform, the susceptibility of Pseudomonas aeruginosa to the antibiotic gentamicin was tested. The result suggests that the proliferation of bacteria is effectively controlled by gentamicin. This study demonstrated a sensitive (one bacterium on single particles) and time-saving (within 2 h) platform with a small sample volume (~0.5 μL) and a low initial bacteria count (50 CFU per droplet ~ 105 CFU/mL) for quantifying the growth of microorganisms depending on Brownian motion. The technique can be applied further to other bacterial strains and increase the success of treatments against infectious diseases in the near future. PMID:26863001

  17. Identification of serum biomarkers for lung cancer using magnetic bead-based SELDI-TOF-MS

    PubMed Central

    Song, Qi-bin; Hu, Wei-guo; Wang, Peng; Yao, Yi; Zeng, Hua-zong

    2011-01-01

    Aim: To identify novel serum biomarkers for lung cancer diagnosis using magnetic bead-based surface-enhanced laser desorption/ionization time-of-flight mass spectrum (SELDI-TOF-MS). Methods: The protein fractions of 121 serum specimens from 30 lung cancer patients, 30 pulmonary tuberculosis patients and 33 healthy controls were enriched using WCX magnetic beads and subjected to SELDI-TOF-MS. The spectra were analyzed using Bio-marker Wizard version 3.1.0 and Biomarker Patterns Software version 5.0. A diagnostic model was constructed with the marker proteins using a linear discrimination analysis method. The validity of this model was tested in a blind test set consisted of 8 randomly selected lung cancer patients, 10 pulmonary tuberculosis patients and 10 healthy volunteers. Results: Seventeen m/z peaks were identified, which were significantly different between the lung cancer group and the control (tuberculosis and healthy control) groups. Among these peaks, the 6445, 9725, 11705, and 15126 m/z peaks were selected by the Biomarker Pattern Software to construct a diagnostic model for lung cancer. This four-peak model established in the training set could discriminate lung cancer patients from non-cancer patients with a sensitivity of 93.3% (28/30) and a specificity of 90.5% (57/63). The diagnostic model showed a high sensitivity (75.0%) and a high specificity (95%) in the blind test validation. Database searching and literature mining indicated that the featured 4 peaks represented chaperonin (M9725), hemoglobin subunit beta (M15335), serum amyloid A (M11548), and an unknown protein. Conclusion: A lung cancer diagnostic model based on bead-based SELDI-TOF-MS has been established for the early diagnosis or differential diagnosis of lung cancers. PMID:22019958

  18. Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation

    PubMed Central

    Rosenbaum, Anna; Bleck, Ellen; Schneider, Matthias; Pongratz, Georg; Vordenbäumen, Stefan

    2016-01-01

    Objective To compare direct and indirect bead-based T-cell isolation followed by magnetic bead-based DNA isolation. Methods T-cells were isolated by direct or indirect selection with magnetic bead coated antbiodies followed by magnetic bead-based automated DNA isolation in 10 healthy subjects. Purity of T-cells, purity of DNA (by A260/A280 ratio measurement) and DNA concentration were assessed. Results Direct and indirect labelling resulted in comparable T-cell purity (93.11±1.47% vs. 94.99±1.54%, p= 0.125) and DNA concentration per cell (50.97±14.15 ng/(mlxcell) vs. 49.53±13.62 ng/(mlxcell), p=0.492), while DNA purity was significantly higher after direct labelling (1.82±0.05 vs. 1.78±0.03, p=0.0488). Conclusions Both direct and indirect magnetic bead-based T-cell selection may be used prior to magnetic bead-based DNA isolation procedures. PMID:27547441

  19. Nanovehicles based Bioassay Labels

    SciTech Connect

    Liu, Guodong; Wang, Jun; Wu, Hong; Lin, Ying-Ying; Lin, Yuehe

    2007-04-01

    In this article, we review recent advances of our group in nanoparticle labels based bioassay. Apoferritin and silica nanoparticles have been used as nanovehicles to load large amount of markers for highly sensitive bioassay. Markers loaded apoferritin, apoferritin-templated metallic phosphate nanoparticles, and poly [guanine] coated silica nanoparticles have been prepared, characterized and used as labels for highly sensitive bioassay of protein and DNA. Dissociation and reconstitution characteristics at different pH as well as the special cavity structure of apoferritin nanovehicle provides a simple and convenient route to prepare versatile nanoparticle labels and avoid the complicated and tedious synthesis process of conventional nanoparticle labels. The optical and electrochemical characteristics of the prepared nanoparticle labels are easily controlled by loading different optical or electrochemical markers. Additionally, the use of apoferritin nanovehicle as template for synthesis of metallic phosphate nanoparticle labels offers fast route to prepare uniform-size metallic nanoparticle labels for electrochemical bioassay and avoids the traditional harsh dissolution conditions to dissolve metallic nanoparticle tags (that is, the strong-acid dissolution of quantum dots and gold nanoparticles) during the stripping analysis step. Silica nanoparticle has also been used as nanovehicle to carry thousands of poly [guanine] tracers, which was used to enhance the oxidation current of Ru(bpy)32+, resulting in enhanced sensitivity of electrochemical immunoassay. The new nanovehicle-based labels have been used for highly sensitive electrochemical detection of DNA and protein biomarkers, such as tumor necrosis factor-alpha (TNF-a). The high sensitivity and selectivity make these labels a useful addition to the armory of nanoparticle-based bioassay. The new nanovehicles based labels hold great promise for multiplex protein and DNA detection and for enhancing the sensitivity

  20. Magnetic Beads-based Bioelectrochemical Immunoassay of Polycyclic Aromatic Hydrocarbons

    SciTech Connect

    Lin, Ying-Ying; Liu, Guodong; Wai, Chien M.; Lin, Yuehe

    2007-07-01

    A simple, rapid, and sensitive bioelectrochemical immunoassay method based on magnetic beads (MBs) has been developed to detect polycyclic aromatic hydrocarbons (PAHs). The principle of this bioassay is based on a direct competitive enzyme-linked immunosorbent assay using PAH-antibody-coated MBs and horseradish peroxidase (HRP)-labeled PAH (HRP-PAH). A magnetic process platform was used to mix and shake the samples during the immunoreactions and to separate free and unbound reagents after the liquid-phase competitive immunoreaction among PAH-antibody-coated MBs, PAH analyte, and HRP-PAH. After a complete immunoassay, the HRP tracers attached to MBs were transferred to a substrate solution containing 3, 3´, 5, 5´- tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2) for electrochemical detection. The voltammetric characteristics of the substrate were investigated, and the reduction peak current of TMB was used to quantify the concentration of PAH. The different parameters, including the amount of HRP-PAH conjugates, the enzyme catalytic reaction time, and the pH of the supporting electrolyte that governs the analytical performance of the immunoassay have been studied in detail and optimized. The detection limit of 50 pg mL-1 was obtained under optimum experimental conditions. The performance of this bioelectrochemical magnetic immunoassay was successfully evaluated with tap water spiked with PAHs, indicating that this convenient and sensitive technique offers great promise for decentralized environmental applications.

  1. Development of a multiplex bead-based assay to monitor dengue virus seroconversion.

    PubMed

    Ng, Kaiting; Connolly, John E

    2014-01-01

    Dengue virus (DENV) envelope protein is responsible for viral attachment to host cells and as such is a target of neutralizing antibody responses. However, the presence of envelope-specific antibodies against a given serotype may contribute to enhanced disease during secondary infection with another serotype. There is a need therefore for a standardized, high-throughput low-volume assay which permits the simultaneous screening of reactivity to multiple DENV serotypes. Here, we describe a method of identifying DENV serotype-specific response in exposed individuals using a multiplexed bead-based immunoassay. The ED3 domain of a specific DENV serotype is cloned into pQEAM containing 6xHIS, TEV protease, and AviTag biotinylation sites. Biotinylated ED3 proteins are expressed in E. coli CVB101 and purified by sequential column fractionation followed by coupling onto fluorescent avidin-coated microspheres. Methods for determining the optimum amount of biotinylated ED3 protein coupled onto the microsphere are described. The assay demonstrates both a high degree of sensitivity and specificity using well-characterized patient plasma samples. The nature of the assay permits further development to include a variety of DENV serotypes and regionally important sub-serotypes. PMID:24696331

  2. A large-area hemispherical perforated bead microarray for monitoring bead based aptamer and target protein interaction

    PubMed Central

    Choi, Jong Seob; Bae, Sunwoong; Kim, Kyung Hoon; Seo, Tae Seok

    2014-01-01

    Herein, we present a large-area 3D hemispherical perforated microwell structure for a bead based bioassay. Such a unique microstructure enables us to perform the rapid and stable localization of the beads at the single bead level and the facile manipulation of the bead capture and retrieval with high speed and efficiency. The fabrication process mainly consisted of three steps: the convex micropatterned nickel (Ni) mold production from the concave micropatterned silicon (Si) wafer, hot embossing on the polymer matrix to generate the concave micropattened acrylate sheet, and reactive ion etching to make the bottom holes. The large-area hemispherical perforated micropatterned acrylate sheet was sandwiched between two polydimethylsiloxane (PDMS) microchannel layers. The bead solution was injected and recovered in the top PDMS microchannel, while the bottom PDMS microchannel was connected with control lines to exert the hydrodynamic force in order to alter the flow direction of the bead solution for the bead capture and release operation. The streptavidin-coated microbead capture was achieved with almost 100% yield within 1 min, and all the beads were retrieved in 10 s. Lysozyme or thrombin binding aptamer labelled microbeads were trapped on the proposed bead microarray, and the in situ fluorescence signal of the bead array was monitored after aptamer-target protein interaction. The protein-aptamer conjugated microbeads were recovered, and the aptamer was isolated for matrix assisted laser desorption/ionization time-of-flight mass spectrometry analysis to confirm the identity of the aptamer. PMID:25587373

  3. Fine-tuning of magnetic and microfluidic viscous forces for specific magnetic bead-based immunocomplex formation

    NASA Astrophysics Data System (ADS)

    Cornaglia, M.; Tekin, H. C.; Lehnert, T.; Gijs, M. A. M.

    2013-08-01

    We investigate the working principle of a novel type of microfluidic sandwich immunoassay, as used for the detection of biomarkers. The heterogeneous assay is based on the specific interactions between an array of functionalized superparamagnetic beads and a flow of secondary superparamagnetic beads that carry the antigens and are simultaneously used as detection labels. We identify the main forces governing the immunoassay performance and develop a combined finite element method/analytical model to predict and control these forces. The clue for the improved assay specificity is in the fine-tuning of inter-bead magnetic dipolar and microfluidic viscous forces, which allows strongly reducing non-specific interactions, while enhancing the specific formation of immunocomplexes. We exploit our theoretical model to explain the enhanced sensitivity of magnetic bead-based immunoassay experiments performed in microfluidic chips.

  4. Opportunities for bead-based multiplex assays in veterinary diagnostic laboratories

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bead based multiplex assays (BBMA) also referred to as Luminex, MultiAnalyte Profiling or cytometric bead array (CBA) assays, are applicable for high throughput, simultaneous detection of multiple analytes in solution (from several, up to 50-500 analytes within a single, small sample volume). Curren...

  5. A bead-based fluorescence immunosensing technique enabled by the integration of Förster resonance energy transfer and optoelectrokinetic concentration.

    PubMed

    Wang, Jhih-Cheng; Ku, Hu-Yao; Shieh, Dar-Bin; Chuang, Han-Sheng

    2016-01-01

    Bead-based immunosensing has been growing as a promising technology in the point-of-care diagnostics due to great flexibility. For dilute samples, functionalized particles can be used to collect dispersed analytes and act as carriers for particle manipulation. To realize rapid and visual immunosensing, Förster resonance energy transfer (FRET) was used herein to ensure only the diabetic biomarker, lipocalin 1, to be detected. The measurement was made in an aqueous droplet sandwiched between two parallel plate electrodes. With an electric field and a focused laser beam applying on the microchip simultaneously, the immunocomplexes in the droplet were further concentrated to enhance the FRET fluorescent signal. The optoelectrokinetic technique, termed rapid electrokinetic patterning (REP), has been proven to be excellent in dynamic and programmable particle manipulation. Therefore, the detection can be complete within several tens of seconds. The lower detection limit of the REP-enabled bead-based diagnosis reached nearly 5 nM. The combinative use of FRET and the optoelectrokinetic technique for the bead-based immunosensing enables a rapid measure to diagnose early stage diseases and dilute analytes. PMID:26865906

  6. Nanoparticle-Based Biosensors and Bioassays

    SciTech Connect

    Liu, Guodong; Wang, Jun; Lin, Yuehe; Wang, Joseph

    2007-10-11

    In this book chapter, we review the recent advances in nanoparticles based bioassay. The nanoparticles include quantum dots, silica nanoparticles and apoferritin nanoparticles. The new nanoparticles-based labels hold great promise for multiplex protein and DNA detection and for enhancing the sensitivity of other bioassays.

  7. A Fully Automatic Method for Gridding Bright Field Images of Bead-Based Microarrays.

    PubMed

    Datta, Abhik; Wai-Kin Kong, Adams; Yow, Kin-Choong

    2016-07-01

    In this paper, a fully automatic method for gridding bright field images of bead-based microarrays is proposed. There have been numerous techniques developed for gridding fluorescence images of traditional spotted microarrays but to our best knowledge, no algorithm has yet been developed for gridding bright field images of bead-based microarrays. The proposed gridding method is designed for automatic quality control during fabrication and assembly of bead-based microarrays. The method begins by estimating the grid parameters using an evolutionary algorithm. This is followed by a grid-fitting step that rigidly aligns an ideal grid with the image. Finally, a grid refinement step deforms the ideal grid to better fit the image. The grid fitting and refinement are performed locally and the final grid is a nonlinear (piecewise affine) grid. To deal with extreme corruptions in the image, the initial grid parameter estimation and grid-fitting steps employ robust search techniques. The proposed method does not have any free parameters that need tuning. The method is capable of identifying the grid structure even in the presence of extreme amounts of artifacts and distortions. Evaluation results on a variety of images are presented. PMID:26011899

  8. Microfluidic bead-based multienzyme-nanoparticle amplification for detection of circulating tumor cells in the blood using quantum dots labels.

    PubMed

    Zhang, He; Fu, Xin; Hu, Jiayi; Zhu, Zhenjun

    2013-05-24

    This study reports the development of a microfluidic bead-based nucleic acid sensor for sensitive detection of circulating tumor cells in blood samples using multienzyme-nanoparticle amplification and quantum dot labels. In this method, the microbeads functionalized with the capture probes and modified electron rich proteins were arrayed within a microfluidic channel as sensing elements, and the gold nanoparticles (AuNPs) functionalized with the horseradish peroxidases (HRP) and DNA probes were used as labels. Hence, two signal amplification approaches are integrated for enhancing the detection sensitivity of circulating tumor cells. First, the large surface area of Au nanoparticle carrier allows several binding events of HRP on each nanosphere. Second, enhanced mass transport capability inherent from microfluidics leads to higher capture efficiency of targets because continuous flow within micro-channel delivers fresh analyte solution to the reaction site which maintains a high concentration gradient differential to enhance mass transport. Based on the dual signal amplification strategy, the developed microfluidic bead-based nucleic acid sensor could discriminate as low as 5 fM (signal-to-noise (S/N)3) of synthesized carcinoembryonic antigen (CEA) gene fragments and showed a 1000-fold increase in detection limit compared to the off-chip test. In addition, using spiked colorectal cancer cell lines (HT29) in the blood as a model system, the detection limit of this chip-based approach was found to be as low as 1 HT29 in 1 mL blood sample. This microfluidic bead-based nucleic acid sensor is a promising platform for disease-related nucleic acid molecules at the lowest level at their earliest incidence. PMID:23663673

  9. Enhancement of preneoplastic lesion yield by Chios Mastic Gum in a rat liver medium-term carcinogenesis bioassay

    SciTech Connect

    Doi, Kenichiro Wei, Min; Kitano, Mitsuaki; Uematsu, Naomi; Inoue, Masayo; Wanibuchi, Hideki

    2009-01-01

    The mastic (Pistacia lentiscus var. chia) tree is native throughout the Mediterranean region and has long proved a source of food additives and medical treatments. To investigate the modifying effects of Chios Mastic Gum on rat liver carcinogenesis, 6-week-old male F344 rats were subjected to the established rat liver medium-term carcinogenesis bioassay (Ito-test). At the commencement, rats (groups 1-4) were intraperitoneally injected with 200 mg/kg body weight of diethylnitrosamine (DEN). After two weeks, mastic was added to CRF (Charles River Formula)-1 powdered basal diet at doses of 0, 0.01, 0.1 and 1% in groups 1-4, respectively. At week 3, all rats were underwent two-thirds partial hepatectomy. The experiment was terminated at week 8. As results show, liver weights were significantly increased in a mastic dose-dependent manner among groups 1-4. The numbers (/cm{sup 2}) and the areas (mm{sup 2}/cm{sup 2}) of glutathione S-transferase placental form (GST-P)-positive cell foci ({>=} 0.2 mm in diameter) were significantly increased in the DEN-1% group compared to the DEN-alone group, along with the average areas per foci and larger-sized foci ({>=} 0.4 mm). 5-Bromo-2'-deoxyuridine (BrdU) + GST-P double-immunohistochemistry showed the highest BrdU-labeling indices within GST-P foci in the DEN-1% group. 8-hydroxydeoxyguanosine (8-OHdG) levels in liver DNA did not vary, while real-time quantitative polymerase chain reaction (PCR) analysis of livers revealed many up- or down-regulated genes in the DEN-1% group. In conclusion, this is the first report to display a promotion potential of Chios Mastic Gum on the formation of preneoplastic lesions in the established rat liver medium-term carcinogenesis bioassay.

  10. Development of a bead-based suspension array for the detection of pathogens in acute respiratory tract infections.

    PubMed

    Chen, Yu-Sheng; Li, Hong-Ru; Zhang, Wei; Hua, Zhi-Dan; Lin, Xiao-Hong; Lin, Meng-Qing; Huang, Wen-Sen; Huang, Li-Ping; Yu, Xiao-Li; Xu, Neng-Luan; Lin, Ming; Xie, Bao-Song; Shen, Xiao-Na; Xie, Jian-Feng; Wang, Yi; Huang, Meng; Wu, Yan-An; Hu, Xin-Lan

    2016-08-01

    We developed a high-throughput bead-based suspension array for simultaneous detection of 20 respiratory tract pathogens in clinical specimens. Pathogen-specific genes were amplified and hybridized to probes coupled to carboxyl-encoded microspheres. Fluorescence intensities generated via the binding of phycoerythrin-conjugated streptavidin with biotin-labeled targets were measured by the Luminex 100 bead-based suspension array system. The bead-based suspension array detected bacteria in a significantly higher number of samples compared to the conventional culture. There was no significant difference in the detection rate of atypical pathogensatypical pathogens or viruses between the bead-based suspension array and real-time PCR. This technology can play a significant role in screening patients with pneumonia. PMID:27190247

  11. Confirming Pseudomonas putida as a reliable bioassay for demonstrating biocompatibility enhancement by solar photo-oxidative processes of a biorecalcitrant effluent.

    PubMed

    García-Ripoll, A; Amat, A M; Arques, A; Vicente, R; Ballesteros Martín, M M; Pérez, J A Sánchez; Oller, I; Malato, S

    2009-03-15

    Experiments based on Vibrio fischeri, activated sludge and Pseudomonas putida have been employed to check variation in the biocompatibility of an aqueous solution of a commercial pesticide, along solar photo-oxidative process (TiO(2) and Fenton reagent). Activated sludge-based experiments have demonstrated a complete detoxification of the solution, although important toxicity is still detected according to the more sensitive V. fischeri assays. In parallel, the biodegradability of organic matter is strongly enhanced, with BOD(5)/COD ratio above 0.8. Bioassays run with P. putida have given similar trends, remarking the convenience of using P. putida culture as a reliable and reproducible method for assessing both toxicity and biodegradability, as a substitute to other more time consuming methods. PMID:18639379

  12. Bead-based suspension array for simultaneous differential detection of five major swine viruses.

    PubMed

    Chen, Ru; Yu, Xiao-Lu; Gao, Xiao-Bo; Xue, Cun-Yi; Song, Chang-Xu; Li, Yan; Cao, Yong-Chang

    2015-01-01

    A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2-22 copies/μL (corresponding to 0.5-6.8 fg/μL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4 % and 4.1 to 7.6 %, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2 h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen. PMID:25557628

  13. Comparison of non-magnetic and magnetic beads in bead-based assays.

    PubMed

    Hansenová Maňásková, Silvie; van Belkum, Alex; Endtz, Hubert P; Bikker, Floris J; Veerman, Enno C I; van Wamel, Willem J B

    2016-09-01

    Multiplex bead-based flow cytometry is an attractive way for simultaneous, rapid and cost-effective analysis of multiple analytes in a single sample. Previously, we developed various bead-based assays using non-magnetic beads coated with Staphylococcus aureus and Streptococcus pneumoniae antigens for the detection of antibodies. Here, we compared the performance of the assay using non-magnetic beads with one based on the newly developed magnetic beads. We optimized the magnetic beads' coupling procedure and antibody detection assays for S. aureus and S. pneumoniae antigens and we measured IgG in human pooled serum against a series of S. aureus and S. pneumoniae-derived antigens in a singleplex and in a multiplex assay, respectively. For the multiplex assay, the comparison between magnetic and non-magnetic beads showed: i) in the majority of the cases (13 of the 17 tested S. pneumoniae antigens) significantly higher Median Fluorescence Intensity (MFI) values, ii) lower detection limits, iii) lower coefficient of variation (CV: 12% vs. 7% for non-magnetic vs. magnetic beads), so lower inter-assay variation and hence higher reproducibility. Magnetic bead coupling is cost effective, as we used 25% of the normal amount of antigen and only 50% of the beads in comparison to the non-magnetic beads. This optimized magnetic-based assay, which combines ease of use with an improved assay performance, allows detection of antibodies with a low titer that are potentially missed with the non-magnetic-based assay. PMID:27296810

  14. Serodiagnosis of Lyme borreliosis with bead based immunoassays using multiplex technology.

    PubMed

    Gerritzen, Andreas; Brandt, Sabine

    2012-04-01

    The serological diagnosis of Lyme borreliosis is accomplished by the detection of IgG and IgM antibodies specific for relevant antigens of the spirochetal pathogen Borrelia burgdorferi. Instead of the usual enzyme immune assay for screening and the Western blot technique for confirmation, bead based multiplex assays with multiple simultaneously performed distinct reactions can provide quick, automatically derived and reliable results in a single run by flow cytometer technology. The broad analytical dynamic range of assay signals and the high sensitivity and specificity of the multiplex formats allow even for a reliable use in CSF based analyses for antibody specificity index in supposed neuroborreliosis. Fluorescence intensity of the bead based reactions can be transformed into quantified values as U/ml, either for each single antigen or summed up for a group of relevant key antigens. Additionally or alternatively distinct reactions of single bead populations can be transformed to Western blot band equivalents. Internal and external quality controls with the multiplex systems show characteristic data equivalent to the conventional assay formats, so that the advantages of the multiplex assays are ready for use in the routine diagnostic laboratory. PMID:22406491

  15. Development of a novel multiplex beads-based assay for autoantibody detection for colorectal cancer diagnosis.

    PubMed

    Villar-Vázquez, Roi; Padilla, Guillermo; Fernández-Aceñero, María Jesús; Suárez, Adolfo; Fuente, Eduardo; Pastor, Carlos; Calero, Miguel; Barderas, Rodrigo; Casal, J Ignacio

    2016-04-01

    Humoral response in cancer patients can be used for early cancer detection. By screening high-density protein microarrays with sera from colorectal cancer (CRC) patients and controls, we identified 16 tumor-associated antigens (TAAs) exhibiting high diagnostic value. This high number of TAAs requires the development of multiplex assays combining different antigens for a faster and more accurate prediction of CRC. Here, we have developed and optimized a bead-based assay using nine selected TAAs and two controls to provide a multiplex test for early CRC diagnosis. We screened a collection of 307 CRC patients' and control sera with the beads assay to identify and validate the best TAA combination for CRC detection. The multiplex bead-based assay exhibited a similar diagnostic performance to detect the humoral response in comparison to multiple ELISA analyses. After multivariate analysis, a panel composed of GTF2B, EDIL3, HCK, PIM1, STK4, and p53, together with gender and age, was identified as the best combination of TAAs for CRC diagnosis, achieving an AUC of 89.7%, with 66% sensitivity at 90.0% fixed specificity. The model was validated using bootstrapping analysis. In summary, we have developed a novel multiplex bead assay that after validation with a larger independent cohort of sera could be utilized in a high-throughput manner for population screening to facilitate the detection of early CRC patients. PMID:26915739

  16. An integrated microfluidic biochemical detection system for protein analysis with magnetic bead-based sampling capabilities.

    PubMed

    Choi, Jin-Woo; Oh, Kwang W; Thomas, Jennifer H; Heineman, William R; Halsall, H Brian; Nevin, Joseph H; Helmicki, Arthur J; Henderson, H Thurman; Ahn, Chong H

    2002-02-01

    This paper presents the development and characterization of an integrated microfluidic biochemical detection system for fast and low-volume immunoassays using magnetic beads, which are used as both immobilization surfaces and bio-molecule carriers. Microfluidic components have been developed and integrated to construct a microfluidic biochemical detection system. Magnetic bead-based immunoassay, as a typical example of biochemical detection and analysis, has been successfully performed on the integrated microfluidic biochemical analysis system that includes a surface-mounted biofilter and electrochemical sensor on a glass microfluidic motherboard. Total time required for an immunoassay was less than 20 min including sample incubation time, and sample volume wasted was less than 50 microl during five repeated assays. Fast and low-volume biochemical analysis has been successfully achieved with the developed biofilter and immunosensor, which is integrated to the microfluidic system. Such a magnetic bead-based biochemical detection system, described in this paper, can be applied to protein analysis systems. PMID:15100857

  17. Five-Antigen Fluorescent Bead-Based Assay for Diagnosis of Lyme Disease.

    PubMed

    Embers, Monica E; Hasenkampf, Nicole R; Barnes, Mary B; Didier, Elizabeth S; Philipp, Mario T; Tardo, Amanda C

    2016-04-01

    The systematically difficult task of diagnosing Lyme disease can be simplified by sensitive and specific laboratory tests. The currently recommended two-tier test for serology is highly specific but falls short in sensitivity, especially in the early acute phase. We previously examined serially collected serum samples fromBorrelia burgdorferi-infected rhesus macaques and defined a combination of antigens that could be utilized for detection of infection at all phases of disease in humans. The fiveB. burgdorferiantigens, consisting of OspC, OspA, DbpA, OppA2, and the C6 peptide, were combined into a fluorescent cytometric bead-based assay for the detection ofB. burgdorferiantigen-specific IgG antibodies. Samples from Lyme disease patients and controls were used to determine the diagnostic value of this assay. Using this sample set, we found that our five-antigen multiplex IgG assay exhibited higher sensitivity (79.5%) than the enzyme immunoassay (EIA) (76.1%), the two-tier test (61.4%), and the C6 peptide enzyme-linked immunosorbent assay (ELISA) (77.2%) while maintaining specificity over 90%. When detection of IgM was added to the bead-based assay, the sensitivity improved to 91%, but at a cost of reduced specificity (78%). These results indicate that the rational combination of antigens in our multiplex assay may offer an improved serodiagnostic test for Lyme disease. PMID:26843487

  18. Porous Bead-Based Diagnostic Platforms: Bridging the Gaps in Healthcare

    PubMed Central

    Chou, Jie; Wong, Jorge; Christodoulides, Nicolaos; Floriano, Pierre N.; Sanchez, Ximena; McDevitt, John

    2012-01-01

    Advances in lab-on-a-chip systems have strong potential for multiplexed detection of a wide range of analytes with reduced sample and reagent volume; lower costs and shorter analysis times. The completion of high-fidelity multiplexed and multiclass assays remains a challenge for the medical microdevice field; as it struggles to achieve and expand upon at the point-of-care the quality of results that are achieved now routinely in remote laboratory settings. This review article serves to explore for the first time the key intersection of multiplexed bead-based detection systems with integrated microfluidic structures alongside porous capture elements together with biomarker validation studies. These strategically important elements are evaluated here in the context of platform generation as suitable for near-patient testing. Essential issues related to the scalability of these modular sensor ensembles are explored as are attempts to move such multiplexed and multiclass platforms into large-scale clinical trials. Recent efforts in these bead sensors have shown advantages over planar microarrays in terms of their capacity to generate multiplexed test results with shorter analysis times. Through high surface-to-volume ratios and encoding capabilities; porous bead-based ensembles; when combined with microfluidic elements; allow for high-throughput testing for enzymatic assays; general chemistries; protein; antibody and oligonucleotide applications. PMID:23202219

  19. Ultrasensitive protein detection: a case for microfluidic magnetic bead-based assays.

    PubMed

    Tekin, H Cumhur; Gijs, Martin A M

    2013-12-21

    We review the use of magnetic micro- and nanoparticles ('magnetic beads') in microfluidic systems for ultrasensitive protein detection. During recent years magnetic beads have been used frequently in immunoassays, either as mobile substrates on which the target antigen is captured, as detection labels, or simultaneously as substrates and labels. The major part of the reviewed work has as application the detection of antibodies or disease biomarkers in serum or of biotoxins from food samples. Several of the most sensitive assays allow protein detection down to fg mL(-1) concentrations. We benchmark the performance of these microfluidic magnetic bead-based assays with the most promising earlier work and with alternative solutions. PMID:24145920

  20. Directed Differentiation of Embryonic Stem Cells Using a Bead-Based Combinatorial Screening Method

    PubMed Central

    Tarunina, Marina; Hernandez, Diana; Johnson, Christopher J.; Rybtsov, Stanislav; Ramathas, Vidya; Jeyakumar, Mylvaganam; Watson, Thomas; Hook, Lilian; Medvinsky, Alexander; Mason, Chris; Choo, Yen

    2014-01-01

    We have developed a rapid, bead-based combinatorial screening method to determine optimal combinations of variables that direct stem cell differentiation to produce known or novel cell types having pre-determined characteristics. Here we describe three experiments comprising stepwise exposure of mouse or human embryonic cells to 10,000 combinations of serum-free differentiation media, through which we discovered multiple novel, efficient and robust protocols to generate a number of specific hematopoietic and neural lineages. We further demonstrate that the technology can be used to optimize existing protocols in order to substitute costly growth factors with bioactive small molecules and/or increase cell yield, and to identify in vitro conditions for the production of rare developmental intermediates such as an embryonic lymphoid progenitor cell that has not previously been reported. PMID:25251366

  1. A bead-based cleavage method for large-scale identification of protease substrates

    PubMed Central

    Wang, Chunli; Ye, Mingliang; Wei, Xiaoluan; Bian, Yangyang; Cheng, Kai; Zou, Hanfa

    2016-01-01

    Proteolysis is a major form of post translational modification which occurs when a protease cleaves peptide bonds in a target protein to modify its activity. Tracking protease substrates is indispensable for understanding its cellular functions. However, it is difficult to directly identify protease substrates because the end products of proteolysis, the cleaved protein fragments, must be identified among the pool of cellular proteins. Here we present a bead-based cleavage approach using immobilized proteome as the screening library to identify protease substrates. This method enables efficient separation of proteolyzed proteins from background protein mixture. Using caspase-3 as the model protease, we have identified 1159 high confident substrates, among which, strikingly, 43.9% of substrates undergo degradation during apoptosis. The huge number of substrates and positive support of in vivo evidence indicate that the BBC method is a powerful tool for protease substrates identification. PMID:26935269

  2. Magnetic bead-based nucleic acid purification kit: Clinical application and performance evaluation in stool specimens.

    PubMed

    Yoon, Jihoon G; Kang, Jin Seok; Hwang, Seung Yong; Song, Jaewoo; Jeong, Seok Hoon

    2016-05-01

    Two different methods - the semi-automated magnetic bead-based kit (SK, Stool DNA/RNA Purification kit®) and the manual membrane column-based kit (QS, QIAamp® DNA Stool Mini kit) - for purifying nucleic acids from clinical stool samples were compared and evaluated. The SK kit was more user-friendly than QS due to the reduced manual processing, partial automation, and short turnaround time with half cost. Furthermore, SK produced high yields in both DNA and RNA extractions but poor purity in RNA extraction. In the assessment of rotavirus and Clostridium difficile infection, both kits had equivalent or more sensitive performance compared with the standard method. Although SK showed some interference and inhibition in nucleic acid extraction, the performance, including the repeatability, linearity, analytical sensitivity, and matrix effect, was sufficient for routine clinical use. PMID:27030641

  3. High Throughput Flow Cytometry Bead-based Multiplex Assay for Identification of Rho GTPase Inhibitors

    PubMed Central

    Surviladze, Zurab; Young, Susan M; Sklar, Larry A

    2015-01-01

    Summary Rho family GTPases and their effector proteins regulate a wide range of cell signaling pathways. In normal physiological conditions their activity is tightly controlled and it is not surprising that their aberrant activation contributes to tumorigenesis or other diseases. For this reason, the identification of small, cell permeable molecules capable of inhibition of Rho GTPases can be extraordinarily useful, particularly if they are specific and act reversibly. Herein we describe a flow cytometric assay, which allows us to measure the activity of six small GTPases simultaneously. GST-tagged small GTPases are bound to six glutathione bead sets each set having a different intensity of red fluorescence at a fixed wavelength. The coated bead sets were washed, combined, and dispensed into 384-well plates with test compounds, and fluorescent-GTP binding was used as the read-out. This multiplex bead-based assay was successfully used for to identify both general and selective inhibitors of Rho family GTPases. PMID:22144280

  4. A novel multiplex bead-based platform highlights the diversity of extracellular vesicles

    PubMed Central

    Koliha, Nina; Wiencek, Yvonne; Heider, Ute; Jüngst, Christian; Kladt, Nikolay; Krauthäuser, Susanne; Johnston, Ian C. D.; Bosio, Andreas; Schauss, Astrid; Wild, Stefan

    2016-01-01

    The surface protein composition of extracellular vesicles (EVs) is related to the originating cell and may play a role in vesicle function. Knowledge of the protein content of individual EVs is still limited because of the technical challenges to analyse small vesicles. Here, we introduce a novel multiplex bead-based platform to investigate up to 39 different surface markers in one sample. The combination of capture antibody beads with fluorescently labelled detection antibodies allows the analysis of EVs that carry surface markers recognized by both antibodies. This new method enables an easy screening of surface markers on populations of EVs. By combining different capture and detection antibodies, additional information on relative expression levels and potential vesicle subpopulations is gained. We also established a protocol to visualize individual EVs by stimulated emission depletion (STED) microscopy. Thereby, markers on single EVs can be detected by fluorophore-conjugated antibodies. We used the multiplex platform and STED microscopy to show for the first time that NK cell–derived EVs and platelet-derived EVs are devoid of CD9 or CD81, respectively, and that EVs isolated from activated B cells comprise different EV subpopulations. We speculate that, according to our STED data, tetraspanins might not be homogenously distributed but may mostly appear as clusters on EV subpopulations. Finally, we demonstrate that EV mixtures can be separated by magnetic beads and analysed subsequently with the multiplex platform. Both the multiplex bead-based platform and STED microscopy revealed subpopulations of EVs that have been indistinguishable by most analysis tools used so far. We expect that an in-depth view on EV heterogeneity will contribute to our understanding of different EVs and functions. PMID:26901056

  5. Magnetic bead-based reverse colorimetric immunoassay strategy for sensing biomolecules.

    PubMed

    Gao, Zhuangqiang; Xu, Mingdi; Hou, Li; Chen, Guonan; Tang, Dianping

    2013-07-16

    A novel reverse colorimetric immunoassay (RCIA) strategy was for the first time designed and utilized for sensitive detection of low-abundance protein (prostate-specific antigen, PSA, used in this case) in biological fluids by coupling highly catalytic efficient catalase with magnetic bead-based peroxidase mimics. To construct such a RCIA system, two nanostructures including magnetic beads and gold nanoparticles were first synthesized and functionalized with anti-PSA capture antibody and catalase/anti-PSA detection antibody, respectively. Thereafter, a specific sandwich-type immunoassay format was employed for determination of PSA by using functional gold nanoparticles as enzymatic bioreactors and anti-PSA-conjugated magnetic beads as a colorimetric developer. The carried catalase, followed by the sandwiched immunocomplex, partially consumed the added hydrogen peroxide in the detection solution, which slowed down the catalytic efficiency of magnetic bead-based peroxidase mimics toward TMB/H2O2, thereby weakening the visible color and decreasing the colorimetric density. Different from conventional colorimetric immunoassay, the RCIA method determined the residual hydrogen peroxide in the substrate after consumption. Under the optimal conditions, the developed RCIA exhibited a wide dynamic range of 0.05-20 ng mL(-1) toward PSA with a detection limit of 0.03 ng mL(-1) at the 3Sblank level. Intra- and interassay coefficients of variation were below 6.1% and 9.3%, respectively. Additionally, the methodology was further validated for the analysis of 12 PSA clinical serum specimens, giving results in good accordance with those obtained by the commercially available enzyme-linked immunosorbent assay (ELISA) method. PMID:23806145

  6. A novel multiplex bead-based platform highlights the diversity of extracellular vesicles.

    PubMed

    Koliha, Nina; Wiencek, Yvonne; Heider, Ute; Jüngst, Christian; Kladt, Nikolay; Krauthäuser, Susanne; Johnston, Ian C D; Bosio, Andreas; Schauss, Astrid; Wild, Stefan

    2016-01-01

    The surface protein composition of extracellular vesicles (EVs) is related to the originating cell and may play a role in vesicle function. Knowledge of the protein content of individual EVs is still limited because of the technical challenges to analyse small vesicles. Here, we introduce a novel multiplex bead-based platform to investigate up to 39 different surface markers in one sample. The combination of capture antibody beads with fluorescently labelled detection antibodies allows the analysis of EVs that carry surface markers recognized by both antibodies. This new method enables an easy screening of surface markers on populations of EVs. By combining different capture and detection antibodies, additional information on relative expression levels and potential vesicle subpopulations is gained. We also established a protocol to visualize individual EVs by stimulated emission depletion (STED) microscopy. Thereby, markers on single EVs can be detected by fluorophore-conjugated antibodies. We used the multiplex platform and STED microscopy to show for the first time that NK cell-derived EVs and platelet-derived EVs are devoid of CD9 or CD81, respectively, and that EVs isolated from activated B cells comprise different EV subpopulations. We speculate that, according to our STED data, tetraspanins might not be homogenously distributed but may mostly appear as clusters on EV subpopulations. Finally, we demonstrate that EV mixtures can be separated by magnetic beads and analysed subsequently with the multiplex platform. Both the multiplex bead-based platform and STED microscopy revealed subpopulations of EVs that have been indistinguishable by most analysis tools used so far. We expect that an in-depth view on EV heterogeneity will contribute to our understanding of different EVs and functions. PMID:26901056

  7. New miRNA labeling method for bead-based quantification

    PubMed Central

    2010-01-01

    Background microRNAs (miRNAs) are small single-stranded non-coding RNAs that act as crucial regulators of gene expression. Different methods have been developed for miRNA expression profiling in order to better understand gene regulation in normal and pathological conditions. miRNAs expression values obtained from large scale methodologies such as microarrays still need a validation step with alternative technologies. Results Here we have applied with an innovative approach, the Luminex® xMAP™ technology validate expression data of differentially expressed miRNAs obtained from high throughput arrays. We have developed a novel labeling system of small RNA molecules (below 200 nt), optimizing the sensitive cloning method for miRNAs, termed miRNA amplification profiling (mRAP). The Luminex expression patterns of three miRNAs (miR-23a, miR-27a and miR-199a) in seven different cell lines have been validated by TaqMan miRNA assay. In all cases, bead-based meas were confirmed by the data obtained by TaqMan and microarray technologies. Conclusions We demonstrate that the measure of individual miRNA by the bead-based method is feasible, high speed, sensitive and low cost. The Luminex® xMAP™ technology also provides flexibility, since the central reaction can be scaled up with additional miRNA capturing beads, allowing validation of many differentially expressed miRNAs obtained from microarrays in a single experiment. We propose this technology as an alternative method to qRT-PCR for validating miRNAs expression data obtained with high-throughput technologies. PMID:20553585

  8. Viral RNA testing and automation on the bead-based CBNE detection microsystem.

    SciTech Connect

    Galambos, Paul C.; Bourdon, Christopher Jay; Farrell, Cara M.; Rossito, Paul; McClain, Jaime L.; Derzon, Mark Steven; Cullor, James Sterling; Rahimian, Kamayar

    2008-09-01

    We developed prototype chemistry for nucleic acid hybridization on our bead-based diagnostics platform and we established an automatable bead handling protocol capable of 50 part-per-billion (ppb) sensitivity. We are working towards a platform capable of parallel, rapid (10 minute), raw sample testing for orthogonal (in this case nucleic acid and immunoassays) identification of biological (and other) threats in a single sensor microsystem. In this LDRD we developed the nucleic acid chemistry required for nucleic acid hybridization. Our goal is to place a non-cell associated RNA virus (Bovine Viral Diarrhea, BVD) on the beads for raw sample testing. This key pre-requisite to showing orthogonality (nucleic acid measurements can be performed in parallel with immunoassay measurements). Orthogonal detection dramatically reduces false positives. We chose BVD because our collaborators (UC-Davis) can supply samples from persistently infected animals; and because proof-of-concept field testing can be performed with modification of the current technology platform at the UC Davis research station. Since BVD is a cattle-prone disease this research dovetails with earlier immunoassay work on Botulinum toxin simulant testing in raw milk samples. Demonstration of BVD RNA detection expands the repertoire of biological macromolecules that can be adapted to our bead-based detection. The resources of this late start LDRD were adequate to partially demonstrate the conjugation of the beads to the nucleic acids. It was never expected to be adequate for a full live virus test but to motivate that additional investment. In addition, we were able to reduce the LOD (Limit of Detection) for the botulinum toxin stimulant to 50 ppb from the earlier LOD of 1 ppm. A low LOD combined with orthogonal detection provides both low false negatives and low false positives. The logical follow-on steps to this LDRD research are to perform live virus identification as well as concurrent nucleic acid and

  9. A cell-based fascin bioassay identifies compounds with potential anti-metastasis or cognition-enhancing functions

    PubMed Central

    Kraft, Robert; Kahn, Allon; Medina-Franco, José L.; Orlowski, Mikayla L.; Baynes, Cayla; López-Vallejo, Fabian; Barnard, Kobus; Maggiora, Gerald M.; Restifo, Linda L.

    2013-01-01

    SUMMARY The actin-bundling protein fascin is a key mediator of tumor invasion and metastasis and its activity drives filopodia formation, cell-shape changes and cell migration. Small-molecule inhibitors of fascin block tumor metastasis in animal models. Conversely, fascin deficiency might underlie the pathogenesis of some developmental brain disorders. To identify fascin-pathway modulators we devised a cell-based assay for fascin function and used it in a bidirectional drug screen. The screen utilized cultured fascin-deficient mutant Drosophila neurons, whose neurite arbors manifest the ‘filagree’ phenotype. Taking a repurposing approach, we screened a library of 1040 known compounds, many of them FDA-approved drugs, for filagree modifiers. Based on scaffold distribution, molecular-fingerprint similarities, and chemical-space distribution, this library has high structural diversity, supporting its utility as a screening tool. We identified 34 fascin-pathway blockers (with potential anti-metastasis activity) and 48 fascin-pathway enhancers (with potential cognitive-enhancer activity). The structural diversity of the active compounds suggests multiple molecular targets. Comparisons of active and inactive compounds provided preliminary structure-activity relationship information. The screen also revealed diverse neurotoxic effects of other drugs, notably the ‘beads-on-a-string’ defect, which is induced solely by statins. Statin-induced neurotoxicity is enhanced by fascin deficiency. In summary, we provide evidence that primary neuron culture using a genetic model organism can be valuable for early-stage drug discovery and developmental neurotoxicity testing. Furthermore, we propose that, given an appropriate assay for target-pathway function, bidirectional screening for brain-development disorders and invasive cancers represents an efficient, multipurpose strategy for drug discovery. PMID:22917928

  10. Effect of enhanced pCO2 levels on the production of DOC and TEP in short-term bioassay experiments

    NASA Astrophysics Data System (ADS)

    MacGilchrist, G. A.; Shi, T.; Tyrrell, T.; Richier, S.; Moore, C. M.; Dumousseaud, C.; Achterberg, E. P.

    2014-03-01

    It has been proposed that increasing levels of pCO2 in the surface ocean will lead to more partitioning of the organic carbon fixed by marine primary production into the dissolved rather than the particulate fraction. This process may result in enhanced accumulation of dissolved organic carbon (DOC) in the surface ocean and/or concurrent accumulation of transparent exopolymer particles (TEP), with important implications for the functioning of the marine carbon cycle. We investigated this in shipboard bioassay experiments that considered the effect of four different pCO2 scenarios (ambient, 550, 750 and 1000 μatm) on unamended natural phytoplankton communities from a range of locations in the northwest European shelf seas. The environmental settings, in terms of nutrient availability, phytoplankton community structure and growth conditions, varied considerably between locations. We did not observe any strong or consistent effect of pCO2 on DOC production. There was a significant but highly variable effect of pCO2 on the production of TEP. In three of the five experiments, variation of TEP production between pCO2 treatments was caused by the effect of pCO2 on phytoplankton growth rather than a direct effect on TEP production. In one of the five experiments, there was evidence of enhanced TEP production at high pCO2 (twice as much production over the 96 h incubation period in the 750 μatm treatment compared with the ambient treatment) independent of indirect effects, as hypothesised by previous studies. Our results suggest that the environmental setting of experiments (community structure, nutrient availability and occurrence of phytoplankton growth) is a key factor determining the TEP response to pCO2 perturbations.

  11. Effect of enhanced pCO2 levels on the production of dissolved organic carbon and transparent exopolymer particles in short-term bioassay experiments

    NASA Astrophysics Data System (ADS)

    MacGilchrist, G. A.; Shi, T.; Tyrrell, T.; Richier, S.; Moore, C. M.; Dumousseaud, C.; Achterberg, E. P.

    2014-07-01

    It has been proposed that increasing levels of pCO2 in the surface ocean will lead to more partitioning of the organic carbon fixed by marine primary production into the dissolved rather than the particulate fraction. This process may result in enhanced accumulation of dissolved organic carbon (DOC) in the surface ocean and/or concurrent accumulation of transparent exopolymer particles (TEPs), with important implications for the functioning of the marine carbon cycle. We investigated this in shipboard bioassay experiments that considered the effect of four different pCO2 scenarios (ambient, 550, 750 and 1000 μatm) on unamended natural phytoplankton communities from a range of locations in the northwest European shelf seas. The environmental settings, in terms of nutrient availability, phytoplankton community structure and growth conditions, varied considerably between locations. We did not observe any strong or consistent effect of pCO2 on DOC production. There was a significant but highly variable effect of pCO2 on the production of TEPs. In three of the five experiments, variation of TEP production between pCO2 treatments was caused by the effect of pCO2 on phytoplankton growth rather than a direct effect on TEP production. In one of the five experiments, there was evidence of enhanced TEP production at high pCO2 (twice as much production over the 96 h incubation period in the 750 μatm treatment compared with the ambient treatment) independent of indirect effects, as hypothesised by previous studies. Our results suggest that the environmental setting of experiments (community structure, nutrient availability and occurrence of phytoplankton growth) is a key factor determining the TEP response to pCO2 perturbations.

  12. A magnetic bead-based ligand binding assay to facilitate human kynurenine 3-monooxygenase drug discovery.

    PubMed

    Wilson, Kris; Mole, Damian J; Homer, Natalie Z M; Iredale, John P; Auer, Manfred; Webster, Scott P

    2015-02-01

    Human kynurenine 3-monooxygenase (KMO) is emerging as an important drug target enzyme in a number of inflammatory and neurodegenerative disease states. Recombinant protein production of KMO, and therefore discovery of KMO ligands, is challenging due to a large membrane targeting domain at the C-terminus of the enzyme that causes stability, solubility, and purification difficulties. The purpose of our investigation was to develop a suitable screening method for targeting human KMO and other similarly challenging drug targets. Here, we report the development of a magnetic bead-based binding assay using mass spectrometry detection for human KMO protein. The assay incorporates isolation of FLAG-tagged KMO enzyme on protein A magnetic beads. The protein-bound beads are incubated with potential binding compounds before specific cleavage of the protein-compound complexes from the beads. Mass spectrometry analysis is used to identify the compounds that demonstrate specific binding affinity for the target protein. The technique was validated using known inhibitors of KMO. This assay is a robust alternative to traditional ligand-binding assays for challenging protein targets, and it overcomes specific difficulties associated with isolating human KMO. PMID:25296660

  13. A Bead-Based Method for Multiplexed Identification and Quantitation of DNA Sequences Using Flow Cytometry

    PubMed Central

    Spiro, Alexander; Lowe, Mary; Brown, Drew

    2000-01-01

    A new multiplexed, bead-based method which utilizes nucleic acid hybridizations on the surface of microscopic polystyrene spheres to identify specific sequences in heterogeneous mixtures of DNA sequences is described. The method consists of three elements: beads (5.6-μm diameter) with oligomer capture probes attached to the surface, three fluorophores for multiplexed detection, and flow cytometry instrumentation. Two fluorophores are impregnated within each bead in varying amounts to create different bead types, each associated with a unique probe. The third fluorophore is a reporter. Following capture of fluorescent cDNA sequences from environmental samples, the beads are analyzed by flow cytometric techniques which yield a signal intensity for each capture probe proportional to the amount of target sequences in the analyte. In this study, a direct hybrid capture assay was developed and evaluated with regard to sequence discrimination and quantitation of abundances. The target sequences (628 to 728 bp in length) were obtained from the 16S/23S intergenic spacer region of microorganisms collected from polluted groundwater at the nuclear waste site in Hanford, Wash. A fluorescence standard consisting of beads with a known number of fluorescent DNA molecules on the surface was developed, and the resolution, sensitivity, and lower detection limit for measuring abundances were determined. The results were compared with those of a DNA microarray using the same sequences. The bead method exhibited far superior sequence discrimination and possesses features which facilitate accurate quantitation. PMID:11010868

  14. Bead-Based Assays for Biodetection: From Flow-Cytometry to Microfluidics

    SciTech Connect

    Ozanich, Richard M.; Antolick, Kathryn C.; Bruckner-Lea, Cindy J.; Bunch, Kyle J.; Dockendorff, Brian P.; Grate, Jay W.; Nash, Michael A.; Tyler, Abby J.

    2009-05-04

    ABSTRACT The potential for the use of biological agents by terrorists is a real threat. Two approaches for detection of biological species will be described: 1) The use of microbead arrays for multiplexed flow cytometry detection of cytokines and botulinum neurotoxin simulant, and 2) a microfluidic platform for capture and separation of different size superparamagnetic nanoparticles followed by on-chip fluorescence detection of the sandwich complex. The methods and automated fluidic systems used for trapping functionalized microbeads will be described. This approach allows sample, assay reagents, and wash solutions to be perfused over a micro-column of beads, resulting in faster and more sensitive assays. The automated fluidic approach resulted in up to five-fold improvements in assay sensitivity/speed as compared to identical assays performed in a typical manual batch mode. A second approach for implementing multiplexed bead-based assays without using flow cytometry detection is currently under development. The goal of the microfluidic-based approach is to achieve rapid (<20 minutes), multiplexed (> 3 bioagents) detection using a simple and low-cost, integrated microfluidic/optical detection platform. Using fiber-optic guided laser-induced fluorescence, assay detection limits were shown to be in the 100’s of picomolar range (10’s of micrograms per liter) for botulinum neurotoxin simulant without any optimization of the microfluidic device or optical detection approach. Video taping magnetic nanoparticle capture and release was used to improve understanding of the process and revealed interesting behavior.

  15. Magnetic beads-based electrochemical immunosensor for monitoring allergenic food proteins.

    PubMed

    Čadková, Michaela; Metelka, Radovan; Holubová, Lucie; Horák, Daniel; Dvořáková, Veronika; Bílková, Zuzana; Korecká, Lucie

    2015-09-01

    Screen-printed platinum electrodes as transducer and magnetic beads as solid phase were combined to develop a particle-based electrochemical immunosensor for monitoring the serious food allergen ovalbumin. The standard arrangement of enzyme-linked immunosorbent assay became the basis for designing the immunosensor. A sandwich-type immunocomplex was formed between magnetic particles functionalized with specific anti-ovalbumin immunoglobulin G and captured ovalbumin molecules, and secondary anti-ovalbumin antibodies conjugated with the enzyme horseradish peroxidase were subsequently added as label tag. The electrochemical signal proportional to the enzymatic reaction of horseradish peroxidase during the reduction of hydrogen peroxide with thionine as electron mediator was measured by linear sweep voltammetry. The newly established method of ovalbumin detection exhibits high sensitivity suitable for quantification in the range of 11 to 222nM and a detection limit of 5nM. Magnetic beads-based assay format using external magnets for rapid and simple separation has been proven to be an excellent basis for electrochemical detection and quantification of food allergens in highly complex sample matrices. PMID:25963896

  16. Bead-based assays for biodetection: from flow-cytometry to microfluidics

    NASA Astrophysics Data System (ADS)

    Ozanich, Richard M., Jr.; Antolick, Kathryn; Bruckner-Lea, Cynthia J.; Bunch, Kyle J.; Dockendorff, Brian P.; Grate, Jay W.; Nash, Michael A.; Tyler, Abby; Warner, Cynthia L.; Warner, Marvin G.

    2009-05-01

    The potential for the use of biological agents by terrorists is a real threat. Two approaches for antibody-based detection of biological species are described in this paper: 1) The use of microbead arrays for multiplexed flow cytometry detection of cytokines and botulinum neurotoxin simulant, and 2) a microfluidic platform for capture and separation of different size superparamagnetic nanoparticles followed by on-chip fluorescence detection of the sandwich complex. These approaches both involve the use of automated fluidic systems for trapping antibody-functionalized microbeads, which allows sample, assay reagents, and wash solutions to be perfused over a micro-column of beads, resulting in faster and more sensitive immunoassays. The automated fluidic approach resulted in up to five-fold improvements in immunoassay sensitivity/speed as compared to identical immunoassays performed in a typical manual batch mode. A second approach for implementing multiplexed bead-based immunoassays without using flow cytometry detection is currently under development. The goal of the microfluidic-based approach is to achieve rapid (<20 minutes), multiplexed (>= 3 bioagents) detection using a simple and low-cost, integrated microfluidic/optical detection platform. Using fiber-optic guided laser-induced fluorescence, assay detection limits were shown to be in the 100's of picomolar range (10's of micrograms per liter) for botulinum neurotoxin simulant without any optimization of the microfluidic device or optical detection approach.

  17. Quantitative Determination of the Probability of Multiple-Motor Transport in Bead-Based Assays.

    PubMed

    Li, Qiaochu; King, Stephen J; Gopinathan, Ajay; Xu, Jing

    2016-06-21

    With their longest dimension typically being less than 100 nm, molecular motors are significantly below the optical-resolution limit. Despite substantial advances in fluorescence-based imaging methodologies, labeling with beads remains critical for optical-trapping-based investigations of molecular motors. A key experimental challenge in bead-based assays is that the number of motors on a bead is not well defined. Particularly for single-molecule investigations, the probability of single- versus multiple-motor events has not been experimentally investigated. Here, we used bead travel distance as an indicator of multiple-motor transport and determined the lower-bound probability of bead transport by two or more motors. We limited the ATP concentration to increase our detection sensitivity for multiple- versus single-kinesin transport. Surprisingly, for all but the lowest motor number examined, our measurements exceeded estimations of a previous model by ≥2-fold. To bridge this apparent gap between theory and experiment, we derived a closed-form expression for the probability of bead transport by multiple motors, and constrained the only free parameter in this model using our experimental measurements. Our data indicate that kinesin extends to ∼57 nm during bead transport, suggesting that kinesin exploits its conformational flexibility to interact with microtubules at highly curved interfaces such as those present for vesicle transport in cells. To our knowledge, our findings provide the first experimentally constrained guide for estimating the probability of multiple-motor transport in optical trapping studies. The experimental approach utilized here (limiting ATP concentration) may be generally applicable to studies in which molecular motors are labeled with cargos that are artificial or are purified from cellular extracts. PMID:27332130

  18. Bead-based microarray immunoassay for lung cancer biomarkers using quantum dots as labels.

    PubMed

    Liu, Lifen; Wu, Simin; Jing, Fengxiang; Zhou, Hongbo; Jia, Chunping; Li, Gang; Cong, Hui; Jin, Qinghui; Zhao, Jianlong

    2016-06-15

    In this study, we developed a multiplex immunoassay system that combines the suspension and planar microarray formats within a single layer of polydimethylsiloxane (PDMS) using soft lithography technology. The suspension format was based on the target proteins forming a sandwich structure between the magnetic beads and the quantum dot (QD) probes through specific antibody-antigen interactions. The planar microarray format was produced by fabricating an array of micro-wells in PDMS. Each micro-well was designed to trap a single microbead and eventually generated a microbead array within the PDMS chamber. The resultant bead-based on-chip assay could be used for simultaneously detecting three lung cancer biomarkers-carcinoembryonic antigen (CEA), fragments of cytokeratin 19 (CYFRA21-1) and neuron-specific enolase (NSE)-in 10 μl of human serum, with a wide linear dynamic range (1.03-111 ng/mL for CEA and CYFRA21-1; 9.26-1000 ng/ml for NSE) and a low detection limit (CEA: 0.19 ng/ml; CYFRA21-1: 0.97 ng/ml; NSE: 0.37 ng/ml; S/N=3). Our micro-well chip does not require complex e-beam lithography or the reactive ion etching process as with existing micro-well systems, which rely on expensive focused ion beam (FIB) milling or optical fiber bundles. Furthermore, the current approach is easy to operate without extra driving equipment such as pumps, and can make parallel detection for multiplexing with rapid binding kinetics, small reagent consumption and low cost. This work has demonstrated the importance of the successful application of on-chip multiplexing sandwich assays for the detection of biomarker proteins. PMID:26852198

  19. A multiplex, bead-based array for profiling plant-derived components in complex food matrixes.

    PubMed

    Ponzoni, Elena; Breviario, Diego; Mautino, Alessandro; Gianì, Silvia; Morello, Laura

    2013-12-01

    Authentication of processed food ingredients is becoming an important issue for customers, and some DNA-based analytical methods have been developed, especially for animal products. As food products typically contain several different ingredients, a current challenge is to increase the multiplexing capacity of DNA-based methods, to develop "all-in-one" assays. Oligonucleotide-coupled, bead-based suspension arrays are sensitive and reproducible multiplex analytical tools. We applied the Multi-Analyte Profile (xMAP™) technology to develop an assay able to concurrently detect five different plant components in mixed flours and in processed feed and food. Capture probes were targeted to species-specific DNA polymorphisms present within the first intron of plant β-tubulin genes, which can be amplified by the tubulin-based polymorphism-amplification method (TBP-PCR). The workflow is very simple and straightforward, consisting of a PCR amplification step with universal primers, followed by the direct hybridization assay. Results are highly reproducible. For each single plant species, the absolute detection limit was as low as one target DNA copy. In complex mixtures of flours derived from seeds or from commercial dry "pasta," relative limits of detection ranged, in weight, from 2% for soybean to less than 0.5% for wheat. The specificity of the capture probes and the high sensitivity of the method allowed the successful determination of the analytical composition of three feeds as well as eleven food samples, such as snacks, biscuits, and pasta. The multiplexing ability of the assay (up to 100 different analytes) provides scalability and flexibility, in response to specific needs. PMID:24190615

  20. Effects of metals in in vitro bioassays.

    PubMed Central

    Sirover, M A

    1981-01-01

    The capacity of in vitro bioassays to detect the potential carcinogenicity of metal compounds is reviewed. The in vitro bioassays discussed include: bacterial reversion analysis to determine the capacity of metal salts to revert Salmonella typhimurium histidine auxotrophs or to revert Escherichia coli WP 2 tryp- to tryptophan prototrophy; examination of the ability of metal salts to preferentially inhibit cell growth in Bacillus subtilis cells deficient in DNA repair pathways; determination of the ability of metal salts to induce resistance to base analogs in mammalian cells; the capacity of metal salts to enhance viral transformation of mammalian cells or to transform cells in the absence of virus; and the ability of metal salts to induce chromosomal aberrations in mammalian cells. Using each of these in vitro bioassays, diverse metal compounds have been identified as potential carcinogens. Furthermore, the use of different compounds of a specific metal may allow a determination of the valence which may be required for carcinogenesis. PMID:7023930

  1. Interlaboratory Comparison of Three Multiplexed Bead-Based Immunoassays for Measuring Serum Antibodies to Pneumococcal Polysaccharides ▿

    PubMed Central

    Whaley, Melissa J.; Rose, Charles; Martinez, Joseph; Laher, Gouri; Sammons, Deborah L.; Smith, Jerry P.; Snawder, John E.; Borrow, Ray; Biagini, Raymond E.; Plikaytis, Brian; Carlone, George M.; Romero-Steiner, Sandra

    2010-01-01

    Serotype-specific IgG, as quantified by a standardized WHO enzyme-linked immunosorbent assay (ELISA), is a serologic end point used to evaluate pneumococcal polysaccharide-based vaccine immunogenicity. Antibodies to each vaccine polysaccharide in licensed multivalent vaccines are quantified separately; this is laborious and consumes serum. We compared three bead-based immunoassays: a commercial assay (xMAP Pneumo14; Luminex) and two in-house assays (of the Health Protection Agency [HPA] and Centers for Disease Control and Prevention [CDC]), using the WHO-recommended standard reference and reference sera (n = 11) from vaccinated adults. Multiple comparisons of the IgG concentrations for seven conjugate vaccine serotypes were performed by sample (percent error), serotype (equivalency testing), and laboratory (concordance correlation coefficient [CCC]). When comparing concentrations by sample, bead-based immunoassays generally yielded higher antibody concentrations than the ELISA and had higher variability for serotypes 6B, 18C, and 23F. None of the three assays met the current WHO recommendation of 75% of sera falling within 40% of the assigned antibody concentrations for all seven serotypes. When compared by serotype, the CDC and HPA tests were equivalent for five of seven serotypes, whereas the Luminex assay was equivalent for four of seven serotypes. When overall mean IgG concentrations were compared by laboratory, a higher level of agreement (CCC close to 1) was found among bead-based immunoassays than between the assays and WHO assignments. When compared to WHO assignments, the HPA assay outperformed the other assays (r = 0.920; CCC = 0.894; coefficient of accuracy = 0.972). Additional testing with sera from immunogenicity studies should demonstrate the applicability of this methodology for vaccine evaluation. PMID:20335434

  2. A novel bead-based assay to detect specific antibody responses against Toxoplasma gondii and Trichinella spiralis simultaneously in sera of experimentally infected swine

    PubMed Central

    2012-01-01

    Background A novel, bead-based flow cytometric assay was developed for simultaneous determination of antibody responses against Toxoplasma gondii and Trichinella spiralis in pig serum. This high throughput screening assay could be an alternative for well known indirect tests like ELISA. One of the advantages of a bead-based assay over ELISA is the possibility to determine multiple specific antibody responses per single sample run facilitated by a series of antigens coupled to identifiable bead-levels. Furthermore, inclusion of a non-coupled bead-level in the same run facilitates the determination of, and correction for non-specific binding. The performance of this bead-based assay was compared to one T. spiralis and three T. gondii ELISAs. For this purpose, sera from T. gondii and T. spiralis experimentally infected pigs were used. With the experimental infection status as gold standard, the area under the curve, Youden Index, sensitivity and specificity were determined through receiver operator curve analysis. Marginal homogeneity and inter-rater agreement between bead-based assay and ELISAs were evaluated using McNemar's Test and Cohen's kappa, respectively. Results Results indicated that the areas under the curve of the bead-based assay were 0.911 and 0.885 for T. gondii and T. spiralis, respectively, while that of the T. gondii ELISAs ranged between 0.837 and 0.930 and the T. spiralis ELISA was 0.879. Bead-based T. gondii assay had a sensitivity of 86% and specificity of 96%, while the ELISAs ranged between 64-84% and 93-99%, respectively. The bead-based T. spiralis assay had a sensitivity of 68% and specificity of 100% while the ELISA scored 72% and 95%, respectively. Marginal homogeneity was found between the T. gondii bead-based test and one of the T. gondii ELISAs. Moreover, in this test combination and between T. spiralis bead-based assay and respective ELISA, an excellent inter-rater agreement was found. When results of samples before expected

  3. Peptidomic analysis of Chinese shrimp ( Fenneropenaeus chinensis) hemolymph by magnetic bead-based MALDI-TOF MS

    NASA Astrophysics Data System (ADS)

    Wang, Baojie; Liu, Mei; Jiang, Keyong; Zhang, Guofan; Wang, Lei

    2013-03-01

    Peptides in shrimp hemolymph play an important role in the innate immune response. Analysis of hemolymph will help to detect and identify potential novel biomarkers of microbial infection. We used magnetic bead-based purification (ClinProt system) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) to characterize shrimp hemolymph peptides. Shrimp serum and plasma were used as the source of samples for comparative analysis, and it was found that serum was more suitable for shrimp hemolymph peptidomic analysis. To screen potential specific biomarkers in serum of immune-challenged shrimps, we applied magnetic bead-based MALDI-TOF MS to serum samples from 10 immune-challenged and 10 healthy shrimps. The spectra were analyzed using FlexAnalysis 3.0 and ClinProTools 2.1 software. Thirteen peptide peaks significantly different between the two groups were selected as candidate biomarkers of lipopolysaccharide (LPS)-infection. The diagnostic model established by genetic algorithm using five of these peaks was able to discriminate LPS-challenged shrimps from healthy control shrimps with a sensitivity of 90% and a specificity of 100%. Our approach in MALDITOF MS-based peptidomics is a powerful tool for screening bioactive peptides or biomarkers derived from hemolymph, and will help to enable a better understanding of the innate immune response of shrimps.

  4. Bioassay for assessing marine contamination

    SciTech Connect

    Lapota, D.; Copeland, H.; Mastny, G.; Rosenberger, D.; Duckworth, D.

    1996-03-01

    The Qwiklite bioassay, developed by the laboratory at NCCOSC, is used as a biological tool to gauge the extent of environmental contamination. Some species of marine phytoplankton produce bioluminescence. The Qwiklite bioassay determines acute response and chronic effects of a wide variety of toxicants upon bioluminescent dinotlagellates by measuring their light output after exposure.

  5. BIOASSAY VESSEL FAILURE ANALYSIS

    SciTech Connect

    Vormelker, P

    2008-09-22

    Two high-pressure bioassay vessels failed at the Savannah River Site during a microwave heating process for biosample testing. Improper installation of the thermal shield in the first failure caused the vessel to burst during microwave heating. The second vessel failure is attributed to overpressurization during a test run. Vessel failure appeared to initiate in the mold parting line, the thinnest cross-section of the octagonal vessel. No material flaws were found in the vessel that would impair its structural performance. Content weight should be minimized to reduce operating temperature and pressure. Outer vessel life is dependent on actual temperature exposure. Since thermal aging of the vessels can be detrimental to their performance, it was recommended that the vessels be used for a limited number of cycles to be determined by additional testing.

  6. Highly Stable Lyophilized Homogeneous Bead-Based Immunoassays for On-Site Detection of Bio Warfare Agents from Complex Matrices.

    PubMed

    Mechaly, Adva; Marx, Sharon; Levy, Orly; Yitzhaki, Shmuel; Fisher, Morly

    2016-06-21

    This study shows the development of dry, highly stable immunoassays for the detection of bio warfare agents in complex matrices. Thermal stability was achieved by the lyophilization of the complete, homogeneous, bead-based immunoassay in a special stabilizing buffer, resulting in a ready-to-use, simple assay, which exhibited long shelf and high-temperature endurance (up to 1 week at 100 °C). The developed methodology was successfully implemented for the preservation of time-resolved fluorescence, Alexa-fluorophores, and horse radish peroxidase-based bead assays, enabling multiplexed detection. The multiplexed assay was successfully implemented for the detection of Bacillus anthracis, botulinum B, and tularemia in complex matrices. PMID:27253489

  7. Development of a novel bead-based 96-well filtration plate competitive immunoassay for the detection of Gentamycin.

    PubMed

    Ho, Tien Yu Jessica; Chan, Chia-Chung; Chan, KinGho; Wang, Yu Chieh; Lin, Jing-Tang; Chang, Cheng-Ming; Chen, Chien-Sheng

    2013-11-15

    We developed a sensitive, simple, inexpensive and rapid bead-based immunoassay platform, composed of liposomal nanovesicle amplification system, Gentamycin sulfate beads and 96-well filtration plates. In the beginning of the assay, Gentamycin sulfate beads, Gentamycin sulfate and Gentamycin specific antibody were incubated in a bottom-sealed 96-well filtration plate. After incubation, washing was done by running washing buffer through the unsealed filtration plate with only gravity and the antibody-Gentamycin bead complexes were retained in the plate. Fluorescent dye-loaded protein G-liposomal nanovesicles were then added to specifically bind to antibodies on the retained beads. After washing unbound nanovesicles, millions of fluorescent dye molecules were released by adding a detergent solution to lyse liposomal nanovesicles. The limit of detection (LOD) of this novel detection platform in TBS and in skim milk were 52.65 ng/mL and 14.16 ng/mL, which are both sufficient for detecting the 200 ng/mL Codex maximum residual level (MRL). The dynamic ranges were both from each of their LODs to 100 μg/mL. The 50% inhibition concentrations (IC50) in TBS and skim milk were 199.66 ng/mL and 360.81 ng/mL, respectively. We also demonstrated the good specificity of this platform by comparing detection results between pure Gentamycin solution and a mixture solution of 6 different antibiotics including Gentamycin in skim milk. The entire assay with 60 samples was conducted within 2h. In sum, this novel biosensing platform not only fulfilled most benefits of magnetic bead-based assays, but also was inexpensive and convenient by replacing the magnetic separation with filtration plate separation. PMID:23728198

  8. Magnetic-bead-based sub-femtomolar immunoassay using resonant Raman scattering signals of ZnS nanoparticles.

    PubMed

    Ding, Yadan; Cong, Tie; Chu, Xueying; Jia, Yan; Hong, Xia; Liu, Yichun

    2016-07-01

    Highly sensitive, specific, and selective immunoassays are of great significance for not only clinical diagnostics but also food safety, environmental monitoring, and so on. Enzyme-linked immunosorbent assays and fluorescence-based and electrochemical immunoassays are important intensively investigated immunoassay techniques. However, they might suffer from low sensitivity or false-positive results. In this work, a simple, reliable, and ultrasensitive magnetic-bead-based immunoassay was performed using biofunctionalized ZnS semiconductor nanocrystals as resonant Raman probes. The resonant Raman scattering of ZnS nanocrystals displays evenly spaced multi-phonon resonant Raman lines with narrow bandwidths and has strong resistance to environmental variation due to the nature of the electron-phonon interaction, thus rendering reliable signal readout in the immunoassays. The superparamagnetic Fe3O4 nanoparticles facilitated greatly the separation, purification, and concentration processes. It is beneficial for both reducing the labor intensity and amplifying the detection signals. The immobilization of antibodies on the surface of magnetic beads, the preparation of resonant Raman probes, and the immunological recognition between the antibody and analyte all occurred in the liquid phase, which minimized the diffusion barriers and boundary layer constraints. All these factors contributed to the ultralow detection limit of human IgG, which was determined to be about 0.5 fM (∼0.08 pg/ml). It is nearly the highest sensitivity obtained for IgG detection. This work shall facilitate the design of nanoplatforms for ultrasensitive detections of proteins, DNAs, bacteria, explosives, and so on. Graphical abstract An ultrasensitive magnetic-bead-based immunoassay was performed using multi-phonon resonant Raman lines of ZnS nanoparticles as detection signals. PMID:27173389

  9. Bioassay Labels Based on Apoferritin Nanovehicles

    SciTech Connect

    Liu, Guodong; Wang, Jun; Lea, Alan S.; Lin, Yuehe

    2006-09-04

    Here we report a nanoparticle label based on apoferritin nanovehicle loaded internally with markers for sensitive electrochemical DNA detection. The central cavity structure, the dissociation and reconstitute properties at different pHs of apoferritin provide a facile method to load and release markers. Hexacynoferrate(III) was used as model marker to load into the cavity of apoferritin protein cage. The loaded nanoparticle surface was functionalized with amino-modified DNA probe. Electrochemical DNA hybridization assay based on the hexacynoferrate loaded apoferritin nanovehicle could detect 23 atmol DNA targets in 50 ul sample solution. The concept could be readily extended to load other redox and fluorescence markers for bioassay applications. The new nanoparticle labels hold great promise for multi-target detection (in connection to nanoparticles loaded with different markers) and for enhancing the sensitivity of other bioassays.

  10. Development of a Multiplexed, Bead-Based Assessment Tool for Rapid Identification and Quantitation of Microorganisms in Field Samples. Final Report

    SciTech Connect

    Lowe, M.; Halden, R.

    2002-10-09

    This was the final report for DOE NABIR grant DE-FG02-01ER63264 (PI Mary Lowe). The grant was entitled ''Development of a Multiplexed Bead-Based Assessment Tool for Rapid Identification and Quantitation of Microorganisms in Field Samples.'' The grant duration was one year. The purpose was to develop a bead-based assay for measuring analyte DNAs in environmental PCR products and to apply the method to a field experiment. The primary experiment was located at the UMTRA Old Rifle site.

  11. Prostaglandins, bioassay and inflammation

    PubMed Central

    Flower, R J

    2006-01-01

    The formation of the British Pharmacological Society coincided almost exactly with a series of ground-breaking studies that ushered in an entirely new field of research – that of lipid mediator pharmacology. For many years following their chemical characterisation, lipids were considered only to be of dietary or structural importance. From the 1930s, all this changed – slowly at first and then more dramatically in the 1970s and 1980s with the emergence of the prostaglandins (PGs), the first intercellular mediators to be clearly derived from lipids, in a dynamic on-demand system. The PGs exhibit a wide range of biological activities that are still being evaluated and their properties underlie the action of one of the world's all-time favourite medicines, aspirin, as well as its more modern congeners. This paper traces the development of the PG field, with particular emphasis on the skilful utilisation of the twin techniques of bioassay and analytical chemistry by U.K. and Swedish scientists, and the intellectual interplay between them that led to the award of a joint Nobel Prize to the principal researchers in the PG field, half a century after the first discovery of these astonishingly versatile mediators. PMID:16402103

  12. EFFECT OF DIFFERENT REGIONS OF AMPLIFIED 16S RDNA ON A PERFORMANCE OF A MULTIPLEXED, BEAD-BASED METHOD FOR ANALYSIS OF DNA SEQUENCES IN ENVIRONMENTAL SAMPLES.

    EPA Science Inventory

    Using a bead-based method for multiplexed analysis of community DNA, the dynamics of aquatic microbial communities can be assessed. Capture probes, specific for a genus or species of bacteria, are attached to the surface of uniquely labeled, microscopic polystyrene beads. Primers...

  13. Development of an automated on-chip bead-based ELISA platform

    PubMed Central

    Campbell, Jennifer; Pollock, Nira; Sharon, Andre; Sauer-Budge, Alexis F.

    2015-01-01

    We present a lab-on-a-chip and associated instrument for heterogeneous enzyme-linked immunosorbent assay (ELISA)-based detection of proteins from liquid samples. The system performs all necessary ELISA steps (starting from antigen incubation) in a quarter of the time required for corresponding plate-based protocols. We have previously described the instrument, which automates fluidic control via remote valve switching and detects fluorescence from reacted substrate, for use in a molecular diagnostics application. The ELISA chip reported here utilizes a high surface area bead bed to enhance capture efficiency and increase the dynamic range of the assay as compared to a standard plate-based ELISA. Its functionality is demonstrated using human IL-10 as a model antigen, but theoretically any sandwich ELISA could be ported onto this “open source platform.” We show that our automated on-chip assays have greater sensitivities than the corresponding standard manual plate-based ELISAs, and that single samples can be assayed in a fraction of the time. PMID:26523155

  14. Dose-response curve of a microfluidic magnetic bead-based surface coverage sandwich assay.

    PubMed

    Cornaglia, Matteo; Trouillon, Raphaël; Tekin, H Cumhur; Lehnert, Thomas; Gijs, Martin A M

    2015-09-25

    Magnetic micro- and nanoparticles ('magnetic beads') have been used to advantage in many microfluidic devices for sensitive antigen (Ag) detection. Today, assays that use as read-out of the signal the number count of immobilized beads on a surface for quantification of a sample's analyte concentration have been among the most sensitive and have allowed protein detection lower than the fgmL(-1) concentration range. Recently, we have proposed in this category a magnetic bead surface coverage assay (Tekin et al., 2013 [1]), in which 'large' (2.8μm) antibody (Ab)-functionalized magnetic beads captured their Ag from a serum and these Ag-carrying beads were subsequently exposed to a surface pattern of fixed 'small' (1.0μm) Ab-coated magnetic beads. When the system was exposed to a magnetic induction field, the magnet dipole attractive interactions between the two bead types were used as a handle to approach both bead surfaces and assist with Ag-Ab immunocomplex formation, while unspecific binding (in absence of an Ag) of a large bead was reduced by exploiting viscous drag flow. The dose-response curve of this type of assay had two remarkable features: (i) its ability to detect an output signal (i.e. bead number count) for very low Ag concentrations, and (ii) an output signal of the assay that was non-linear with respect to Ag concentration. We explain here the observed dose-response curves and show that the type of interactions and the concept of our assay are in favour of detecting the lowest analyte concentrations (where typically either zero or one Ag is carried per large bead), while higher concentrations are less efficiently detected. We propose a random walk process for the Ag-carrying bead over the magnetic landscape of small beads and this model description explains the enhanced overall capture probability of this assay and its particular non-linear dose response curves. PMID:25817550

  15. Rapid magnetic bead based sample preparation for automated and high throughput N-glycan analysis of therapeutic antibodies.

    PubMed

    Váradi, Csaba; Lew, Clarence; Guttman, András

    2014-06-17

    Full automation to enable high throughput N-glycosylation profiling and sequencing with good reproducibility is vital to fulfill the contemporary needs of the biopharmaceutical industry and requirements of national regulatory agencies. The most prevalently used glycoanalytical methods of capillary electrophoresis and hydrophilic interaction liquid chromatography, while very efficient, both necessitate extensive sample preparation and cleanup, including glycoprotein capture, N-glycan release, fluorescent derivatization, purification, and preconcentration steps during the process. Currently used protocols to fulfill these tasks require multiple centrifugation and vacuum-centrifugation steps, making liquid handling robot mediated automated sample preparation difficult and expensive. In this paper we report on a rapid magnetic bead based sample preparation approach that enables full automation including all the process phases just in a couple of hours without requiring any centrifugation and/or vacuum centrifugation steps. This novel protocol has been compared to conventional glycan sample preparation strategies using standard glycoproteins (IgG, fetuin, and RNase B) and featured rapid processing time, high release and labeling efficiency, good reproducibility, and the potential of easy automation. PMID:24909945

  16. Development of a Multiplexed Bead-Based Suspension Array for the Detection and Discrimination of Pospiviroid Plant Pathogens

    PubMed Central

    van Brunschot, Sharon L.; Bergervoet, Jan H. W.; Pagendam, Daniel E.; de Weerdt, Marjanne; Geering, Andrew D. W.; Drenth, André; van der Vlugt, René A. A.

    2014-01-01

    Efficient and reliable diagnostic tools for the routine indexing and certification of clean propagating material are essential for the management of pospiviroid diseases in horticultural crops. This study describes the development of a true multiplexed diagnostic method for the detection and identification of all nine currently recognized pospiviroid species in one assay using Luminex bead-based suspension array technology. In addition, a new data-driven, statistical method is presented for establishing thresholds for positivity for individual assays within multiplexed arrays. When applied to the multiplexed array data generated in this study, the new method was shown to have better control of false positives and false negative results than two other commonly used approaches for setting thresholds. The 11-plex Luminex MagPlex-TAG pospiviroid array described here has a unique hierarchical assay design, incorporating a near-universal assay in addition to nine species-specific assays, and a co-amplified plant internal control assay for quality assurance purposes. All assays of the multiplexed array were shown to be 100% specific, sensitive and reproducible. The multiplexed array described herein is robust, easy to use, displays unambiguous results and has strong potential for use in routine pospiviroid indexing to improve disease management strategies. PMID:24404188

  17. Development of a multiplexed bead-based immunoassay for the simultaneous detection of antibodies to 17 pneumococcal proteins.

    PubMed

    Shoma, S; Verkaik, N J; de Vogel, C P; Hermans, P W M; van Selm, S; Mitchell, T J; van Roosmalen, M; Hossain, S; Rahman, M; Endtz, H Ph; van Wamel, W J B; van Belkum, A

    2011-04-01

    Presently, several pneumococcal proteins are being evaluated as potential vaccine candidates. Here, we gather novel insights in the immunogenicity of PLY, PsaA, PspA, PspC, NanA, Hyl, PpmA, SlrA, Eno, IgA1-protease, PdBD, BVH-3, SP1003, SP1633, SP1651, SP0189 and SP0376. We developed a multiplex bead-based immunoassay (xMAP(®) Technology, Luminex Corporation) to simultaneously quantify antibodies against these 17 pneumococcal proteins in serum. The median fluorescence intensity (MFI) values obtained for human pooled serum with the multiplex assay were between 82% and 111% (median 94%) of those obtained with the singleplex assays. For IgG, the coefficient of variation (CV) in serum ranged from 2% to 9%, for IgA, the CV ranged from 3% to 14% and for IgM, the CV ranged from 11% to 15%. Using this immunoassay, we showed that anti-pneumococcal antibody levels exhibited extensive inter-individual variability in young children suffering from invasive pneumococcal disease. All proteins, including the proteins with, as yet, unknown function, were immunogenic. In conclusion, the multiplex Streptococcus pneumoniae immunoassay based on proteins is reproducible. This assay can be used to monitor anti-S. pneumoniae antibody responses in a material- and time-saving manner. PMID:21086008

  18. Automated microfluidic platform of bead-based electrochemical immunosensor integrated with bioreactor for continual monitoring of cell secreted biomarkers

    PubMed Central

    Riahi, Reza; Shaegh, Seyed Ali Mousavi; Ghaderi, Masoumeh; Zhang, Yu Shrike; Shin, Su Ryon; Aleman, Julio; Massa, Solange; Kim, Duckjin; Dokmeci, Mehmet Remzi; Khademhosseini, Ali

    2016-01-01

    There is an increasing interest in developing microfluidic bioreactors and organs-on-a-chip platforms combined with sensing capabilities for continual monitoring of cell-secreted biomarkers. Conventional approaches such as ELISA and mass spectroscopy cannot satisfy the needs of continual monitoring as they are labor-intensive and not easily integrable with low-volume bioreactors. This paper reports on the development of an automated microfluidic bead-based electrochemical immunosensor for in-line measurement of cell-secreted biomarkers. For the operation of the multi-use immunosensor, disposable magnetic microbeads were used to immobilize biomarker-recognition molecules. Microvalves were further integrated in the microfluidic immunosensor chip to achieve programmable operations of the immunoassay including bead loading and unloading, binding, washing, and electrochemical sensing. The platform allowed convenient integration of the immunosensor with liver-on-chips to carry out continual quantification of biomarkers secreted from hepatocytes. Transferrin and albumin productions were monitored during a 5-day hepatotoxicity assessment in which human primary hepatocytes cultured in the bioreactor were treated with acetaminophen. Taken together, our unique microfluidic immunosensor provides a new platform for in-line detection of biomarkers in low volumes and long-term in vitro assessments of cellular functions in microfluidic bioreactors and organs-on-chips. PMID:27098564

  19. Automated microfluidic platform of bead-based electrochemical immunosensor integrated with bioreactor for continual monitoring of cell secreted biomarkers.

    PubMed

    Riahi, Reza; Shaegh, Seyed Ali Mousavi; Ghaderi, Masoumeh; Zhang, Yu Shrike; Shin, Su Ryon; Aleman, Julio; Massa, Solange; Kim, Duckjin; Dokmeci, Mehmet Remzi; Khademhosseini, Ali

    2016-01-01

    There is an increasing interest in developing microfluidic bioreactors and organs-on-a-chip platforms combined with sensing capabilities for continual monitoring of cell-secreted biomarkers. Conventional approaches such as ELISA and mass spectroscopy cannot satisfy the needs of continual monitoring as they are labor-intensive and not easily integrable with low-volume bioreactors. This paper reports on the development of an automated microfluidic bead-based electrochemical immunosensor for in-line measurement of cell-secreted biomarkers. For the operation of the multi-use immunosensor, disposable magnetic microbeads were used to immobilize biomarker-recognition molecules. Microvalves were further integrated in the microfluidic immunosensor chip to achieve programmable operations of the immunoassay including bead loading and unloading, binding, washing, and electrochemical sensing. The platform allowed convenient integration of the immunosensor with liver-on-chips to carry out continual quantification of biomarkers secreted from hepatocytes. Transferrin and albumin productions were monitored during a 5-day hepatotoxicity assessment in which human primary hepatocytes cultured in the bioreactor were treated with acetaminophen. Taken together, our unique microfluidic immunosensor provides a new platform for in-line detection of biomarkers in low volumes and long-term in vitro assessments of cellular functions in microfluidic bioreactors and organs-on-chips. PMID:27098564

  20. Automated microfluidic platform of bead-based electrochemical immunosensor integrated with bioreactor for continual monitoring of cell secreted biomarkers

    NASA Astrophysics Data System (ADS)

    Riahi, Reza; Shaegh, Seyed Ali Mousavi; Ghaderi, Masoumeh; Zhang, Yu Shrike; Shin, Su Ryon; Aleman, Julio; Massa, Solange; Kim, Duckjin; Dokmeci, Mehmet Remzi; Khademhosseini, Ali

    2016-04-01

    There is an increasing interest in developing microfluidic bioreactors and organs-on-a-chip platforms combined with sensing capabilities for continual monitoring of cell-secreted biomarkers. Conventional approaches such as ELISA and mass spectroscopy cannot satisfy the needs of continual monitoring as they are labor-intensive and not easily integrable with low-volume bioreactors. This paper reports on the development of an automated microfluidic bead-based electrochemical immunosensor for in-line measurement of cell-secreted biomarkers. For the operation of the multi-use immunosensor, disposable magnetic microbeads were used to immobilize biomarker-recognition molecules. Microvalves were further integrated in the microfluidic immunosensor chip to achieve programmable operations of the immunoassay including bead loading and unloading, binding, washing, and electrochemical sensing. The platform allowed convenient integration of the immunosensor with liver-on-chips to carry out continual quantification of biomarkers secreted from hepatocytes. Transferrin and albumin productions were monitored during a 5-day hepatotoxicity assessment in which human primary hepatocytes cultured in the bioreactor were treated with acetaminophen. Taken together, our unique microfluidic immunosensor provides a new platform for in-line detection of biomarkers in low volumes and long-term in vitro assessments of cellular functions in microfluidic bioreactors and organs-on-chips.

  1. Facile Fabrication of a Silver Nanoparticle Immersed, Surface-Enhanced Raman Scattering Imposed Paper Platform through Successive Ionic Layer Absorption and Reaction for On-Site Bioassays.

    PubMed

    Kim, Wansun; Kim, Yeon-Hee; Park, Hun-Kuk; Choi, Samjin

    2015-12-23

    We introduce a novel, facile, rapid, low-cost, highly reproducible, and power-free synthesizable fabrication method of paper-based silver nanoparticle (AgNP) immersed surface-enhanced Raman scattering (SERS) platform, known as the successive ionic layer absorption and reaction (SILAR) method. The rough and porous properties of the paper led to direct synthesis of AgNPs on the surface as well as in the paper due to capillary effects, resulting in improved plasmon coupling with interparticles and interlayers. The proposed SERS platform showed an enhancement factor of 1.1 × 10(9), high reproducibility (relative standard deviation of 4.2%), and 10(-12) M rhodamine B highly sensitive detection limit by optimizing the SILAR conditions including the concentration of the reactive solution (20/20 mM/mM AgNO3/NaBH4) and the number of SILAR cycles (six). The applicability of the SERS platform was evaluated using two samples including human cervical fluid for clinical diagnosis of human papillomavirus (HPV) infection, associated with cervical cancer, and a malachite green (MG) solution for fungicide and parasiticide in aquaculture, associated with human carcinogenesis. The AgNP-immersed SERS-functionalized platform using the SILAR technique allowed for high chemical structure sensitivity without additional tagging or chemical modification, making it a good alternative for early clinical diagnosis of HPV infection and detection of MG-activated human carcinogenesis. PMID:26619139

  2. An automated microfluidic system for single-stranded DNA preparation and magnetic bead-based microarray analysis

    PubMed Central

    Wang, Shuaiqin; Sun, Yujia; Liu, Yan; Xiang, Guangxin; Wang, Lei; Cheng, Jing; Liu, Peng

    2015-01-01

    We present an integrated microfluidic device capable of performing single-stranded DNA (ssDNA) preparation and magnetic bead-based microarray analysis with a white-light detection for detecting mutations that account for hereditary hearing loss. The entire operation process, which includes loading of streptavidin-coated magnetic beads (MBs) and biotin-labeled polymerase chain reaction products, active dispersion of the MBs with DNA for binding, alkaline denaturation of DNA, dynamic hybridization of the bead-labeled ssDNA to a tag array, and white-light detection, can all be automatically accomplished in a single chamber of the microchip, which was operated on a self-contained instrument with all the necessary components for thermal control, fluidic control, and detection. Two novel mixing valves with embedded polydimethylsiloxane membranes, which can alternately generate a 3-μl pulse flow at a peak rate of around 160 mm/s, were integrated into the chip for thoroughly dispersing magnetic beads in 2 min. The binding efficiency of biotinylated oligonucleotides to beads was measured to be 80.6% of that obtained in a tube with the conventional method. To critically test the performance of this automated microsystem, we employed a commercial microarray-based detection kit for detecting nine mutation loci that account for hereditary hearing loss. The limit of detection of the microsystem was determined as 2.5 ng of input K562 standard genomic DNA using this kit. In addition, four blood samples obtained from persons with mutations were all correctly typed by our system in less than 45 min per run. The fully automated, “amplicon-in-answer-out” operation, together with the white-light detection, makes our system an excellent platform for low-cost, rapid genotyping in clinical diagnosis. PMID:25825617

  3. An automated microfluidic system for single-stranded DNA preparation and magnetic bead-based microarray analysis.

    PubMed

    Wang, Shuaiqin; Sun, Yujia; Gan, Wupeng; Liu, Yan; Xiang, Guangxin; Wang, Dong; Wang, Lei; Cheng, Jing; Liu, Peng

    2015-03-01

    We present an integrated microfluidic device capable of performing single-stranded DNA (ssDNA) preparation and magnetic bead-based microarray analysis with a white-light detection for detecting mutations that account for hereditary hearing loss. The entire operation process, which includes loading of streptavidin-coated magnetic beads (MBs) and biotin-labeled polymerase chain reaction products, active dispersion of the MBs with DNA for binding, alkaline denaturation of DNA, dynamic hybridization of the bead-labeled ssDNA to a tag array, and white-light detection, can all be automatically accomplished in a single chamber of the microchip, which was operated on a self-contained instrument with all the necessary components for thermal control, fluidic control, and detection. Two novel mixing valves with embedded polydimethylsiloxane membranes, which can alternately generate a 3-μl pulse flow at a peak rate of around 160 mm/s, were integrated into the chip for thoroughly dispersing magnetic beads in 2 min. The binding efficiency of biotinylated oligonucleotides to beads was measured to be 80.6% of that obtained in a tube with the conventional method. To critically test the performance of this automated microsystem, we employed a commercial microarray-based detection kit for detecting nine mutation loci that account for hereditary hearing loss. The limit of detection of the microsystem was determined as 2.5 ng of input K562 standard genomic DNA using this kit. In addition, four blood samples obtained from persons with mutations were all correctly typed by our system in less than 45 min per run. The fully automated, "amplicon-in-answer-out" operation, together with the white-light detection, makes our system an excellent platform for low-cost, rapid genotyping in clinical diagnosis. PMID:25825617

  4. Novel multiplex bead-based assay for detection of IDH1 and IDH2 mutations in myeloid malignancies.

    PubMed

    Shivarov, Velizar; Ivanova, Milena; Hadjiev, Evgueniy; Naumova, Elissaveta

    2013-01-01

    Isocitrate dehydrogenase 1 and 2 (IDH) mutations are frequently found in various cancer types such as gliomas, chondrosarcomas and myeloid malignancies. Their molecular detection has recently gained wide recognition in the diagnosis and prognosis of these neoplasms. For that purpose various molecular approaches have been used but a universally accepted method is still lacking. In this study we aimed to develop a novel bead-based liquid assay using Locked nucleic acids (LNA)-modified oligonucleotide probes for multiplexed detection of the most frequent IDH1 (p.R132C, p.R132G, p.R132H, p.R132L, p.R132S) and IDH2 (p.R140Q, p.R172K) mutations. The method includes four steps: 1) PCR amplification of the targeted fragments with biotinylated primers; 2) Direct hybridization to barcoded microbeads with specific LNA-modified oligonucleotide probes; 3) Incubation with phycoerythrin coupled streptavidin; 4) Acquisition of fluorescent intensities of each set of beads on a flow platform (LuminexCorp., USA). We tested the performance of the assay on both artificial plasmid constructs and on clinical samples from 114 patients with known or suspected myeloid malignancies. The method appeared to be superior to direct sequencing having a much higher sensitivity of 2.5% mutant alleles. Applying this method to patients' samples we identified a total of 9 mutations (one IDH1 p.R132C, seven IDH2 p.R140Q and one IDH2 p.R172K). In conclusion, this method could be successfully implemented in the diagnostic work-up for various tumors known to harbor IDH1/2 mutations (e.g. myeloid malignancies, gliomas, etc.). International initiatives are needed to validate the different existing methods for detection of IDH1/2 mutations in clinical settings. PMID:24098815

  5. Novel Multiplex Bead-Based Assay for Detection of IDH1 and IDH2 Mutations in Myeloid Malignancies

    PubMed Central

    Hadjiev, Evgueniy; Naumova, Elissaveta

    2013-01-01

    Isocitrate dehydrogenase 1 and 2 (IDH) mutations are frequently found in various cancer types such as gliomas, chondrosarcomas and myeloid malignancies. Their molecular detection has recently gained wide recognition in the diagnosis and prognosis of these neoplasms. For that purpose various molecular approaches have been used but a universally accepted method is still lacking. In this study we aimed to develop a novel bead-based liquid assay using Locked nucleic acids (LNA)-modified oligonucleotide probes for multiplexed detection of the most frequent IDH1 (p.R132C, p.R132G, p.R132H, p.R132L, p.R132S) and IDH2 (p.R140Q, p.R172K) mutations. The method includes four steps: 1) PCR amplification of the targeted fragments with biotinylated primers; 2) Direct hybridization to barcoded microbeads with specific LNA-modified oligonucleotide probes; 3) Incubation with phycoerythrin coupled streptavidin; 4) Acquisition of fluorescent intensities of each set of beads on a flow platform (LuminexCorp., USA). We tested the performance of the assay on both artificial plasmid constructs and on clinical samples from 114 patients with known or suspected myeloid malignancies. The method appeared to be superior to direct sequencing having a much higher sensitivity of 2.5% mutant alleles. Applying this method to patients' samples we identified a total of 9 mutations (one IDH1 p.R132C, seven IDH2 p.R140Q and one IDH2 p.R172K). In conclusion, this method could be successfully implemented in the diagnostic work-up for various tumors known to harbor IDH1/2 mutations (e.g. myeloid malignancies, gliomas, etc.). International initiatives are needed to validate the different existing methods for detection of IDH1/2 mutations in clinical settings. PMID:24098815

  6. Sediment bioassays with oyster larvae

    SciTech Connect

    Chapman, P.M.; Morgan, J.D.

    1983-10-01

    Tests with naturally-occurring sediments are rare and sediment testing methodology is not standardized. The authors present a simple methodology for undertaking sediment bioassays with oyster larvae, and present data from a recent study to prove the utility of this method.

  7. UNIFYING SCALER FOR BIOASSAY TESTS

    EPA Science Inventory

    An extensive set of interlaboratory root bioassay data was unified using centroids of individual tests as scalers. It is shown that the dose response obeys a first order differential equation with the constant of the equation related to the sensitivity of the dose response relati...

  8. Environmental monitoring using genetic bioassays

    SciTech Connect

    Lewtas, J.

    1989-01-01

    Environmental monitoring has evolved over the last ten years toward providing data more useful for exposure and risk assessment. The objective of many monitoring studies in the 1960s and 1970s was to monitor concentrations of pollutants including environmental mutagens at ambient locations, such as roof tops and in large bodies of water, where the pollutants would be well mixed and represent a homogeneous sample. In the 1980s, a number of studies focused on assessing the emission of mutagens from various sources. Now the emphasis has shifted to monitoring human exposure to environmental mutagens and to understanding which sources and factors lead to increased exposure and potential cancer risk. The chapter briefly reviews advances in genetic bioassay methods for environmental monitoring and focuses on approaches to integrating genetic bioassay methods with environmental-monitoring studies.

  9. Soil bioassays as tools for sludge compost quality assessment

    SciTech Connect

    Domene, Xavier; Sola, Laura; Ramirez, Wilson; Alcaniz, Josep M.; Andres, Pilar

    2011-03-15

    Composting is a waste management technology that is becoming more widespread as a response to the increasing production of sewage sludge and the pressure for its reuse in soil. In this study, different bioassays (plant germination, earthworm survival, biomass and reproduction, and collembolan survival and reproduction) were assessed for their usefulness in the compost quality assessment. Compost samples, from two different composting plants, were taken along the composting process, which were characterized and submitted to bioassays (plant germination and collembolan and earthworm performance). Results from our study indicate that the noxious effects of some of the compost samples observed in bioassays are related to the low organic matter stability of composts and the enhanced release of decomposition endproducts, with the exception of earthworms, which are favored. Plant germination and collembolan reproduction inhibition was generally associated with uncomposted sludge, while earthworm total biomass and reproduction were enhanced by these materials. On the other hand, earthworm and collembolan survival were unaffected by the degree of composting of the wastes. However, this pattern was clear in one of the composting procedures assessed, but less in the other, where the release of decomposition endproducts was lower due to its higher stability, indicating the sensitivity and usefulness of bioassays for the quality assessment of composts.

  10. Plasmonically amplified fluorescence bioassay with microarray format

    NASA Astrophysics Data System (ADS)

    Gogalic, S.; Hageneder, S.; Ctortecka, C.; Bauch, M.; Khan, I.; Preininger, Claudia; Sauer, U.; Dostalek, J.

    2015-05-01

    Plasmonic amplification of fluorescence signal in bioassays with microarray detection format is reported. A crossed relief diffraction grating was designed to couple an excitation laser beam to surface plasmons at the wavelength overlapping with the absorption and emission bands of fluorophore Dy647 that was used as a label. The surface of periodically corrugated sensor chip was coated with surface plasmon-supporting gold layer and a thin SU8 polymer film carrying epoxy groups. These groups were employed for the covalent immobilization of capture antibodies at arrays of spots. The plasmonic amplification of fluorescence signal on the developed microarray chip was tested by using interleukin 8 sandwich immunoassay. The readout was performed ex situ after drying the chip by using a commercial scanner with high numerical aperture collecting lens. Obtained results reveal the enhancement of fluorescence signal by a factor of 5 when compared to a regular glass chip.

  11. Medium-term bioassays for carcinogenicity of chemical mixtures.

    PubMed Central

    Ito, N; Imaida, K; Hirose, M; Shirai, T

    1998-01-01

    Carcinogenic effects of chemical mixtures were examined with a medium-term liver bioassay for carcinogens or a multiorgan medium-term bioassay using male F344 rats. In the medium-term liver bioassay, rats were initially treated with diethylnitrosamine (DEN) at 200 mg/kg body weight, i.p.; after 2 weeks they received chemical mixtures such as 10 different heterocyclic amines at one-tenth or one-hundredth the dose levels used in carcinogenicity studies and the mixtures of 20 different pesticides, each at acceptable daily intake (ADI) levels or a mixture of 100 times ADI levels. All animals were subjected to two-thirds partial hepatectomy at week 3 and were sacrificed at week 8. The number and areas of glutathione S-transferase placental form (GST-P) positive foci (preneoplastic lesions in the liver) were compared between respective groups. When 10 heterocyclic amines were mixed in the diet at one-tenth dose level, clear synergism was observed, but no combined effects were evident with the one-hundredth dose levels. In the pesticide experiment, treatment of rats with the 20-pesticide mixture at the ADI dose level did not enhance GST-P-positive foci. In contrast, a mixture of 100 times the ADI significantly increased those values. In a multiorgan bioassay of 28 weeks, mixtures of 40 high-volume compounds and 20 pesticides (suspected carcinogens) added together at their respective ADI levels did not enhance carcinogenesis in any organs initiated by five different carcinogens (DEN, N-methylnitrosourea, dimethylhydrazine, N-butyl-N-(4-hydroxybutyl)nitrosamine, and dihydroxy-di-n-propylnitrosamine) in combination. The combination effect of low dietary levels of five antioxidants, butylated hydroxyanisole, caffeic acid, sesamol, 4-methoxyphenol, and catechol, were also examined using the multiorgan bioassay. The incidence of forestomach papillomas was significantly increased only in the combination group and the results indicate that combination of the five antioxidants can

  12. Comparison of autoantibody specificities between traditional and bead-based assays in a large, diverse collection of SLE patients and family members

    PubMed Central

    Bruner, Benjamin F.; Guthridge, Joel M.; Lu, Rufei; Vidal, Gabriel; Kelly, Jennifer A.; Robertson, Julie M.; Kamen, Diane L.; Gilkeson, Gary S.; Neas, Barbara R.; Reichlin, Morris; Scofield, R. Hal; Harley, John B.; James, Judith A.

    2012-01-01

    Objective The replacement of standard immunofluorescence anti-nuclear antibody (ANA) methods with bead-based assays is a new clinical option. A large, multi-racial cohort of SLE patients, blood relatives and unaffected control individuals was evaluated for familial aggregation and subset clustering of autoantibodies by high-throughput serum screening technology and traditional methods. Methods Serum samples (1,540 SLE patients, 1,127 unaffected relatives, and 906 healthy, population-based controls) were analyzed for SLE autoantibodies using a bead-based assay, immunofluorescence, and immunodiffusion. Autoantibody prevalence, disease sensitivity, clustering, and association with standard immunodiffusion results were evaluated. Results ANA frequency in SLE patient sera were 89%, 73%, and 67% by BioPlex 2200 and 94%, 84%, and 86% by immunofluorescence in African-American, Hispanic, and European-American patients respectively. 60kD Ro, La, Sm, nRNP A, and ribosomal P prevalence were compared across assays, with sensitivities ranging from 0.92 to 0.83 and specificities ranging from 0.90 to 0.79. Cluster autoantibody analysis showed association of three subsets: 1) 60kD Ro, 52kD Ro and La, 2) spliceosomal proteins, and 3) dsDNA, chromatin, and ribosomal P. Familial aggregation of Sm/RNP, ribosomal P, and 60kD Ro in SLE patient sibling pairs was observed (p ≤ 0.004). Simplex pedigree patients had a greater prevalence for dsDNA (p=0.0003) and chromatin (p=0.005) autoantibodies than multiplex patients. Conclusion ANA frequencies detected by a bead-based assay are lower in European-American SLE patients compared to immunofluorescence. These assays have strong positive predictive values across racial groups, provide useful information for clinical care, and provide unique insights to familial aggregation and autoantibody clustering. PMID:23112091

  13. Nanomaterial-Based Electrochemical Biosensors and Bioassays

    SciTech Connect

    Liu, Guodong; Mao, Xun; Gurung, Anant; Baloda, Meenu; Lin, Yuehe; He, Yuqing

    2010-08-31

    This book chapter summarizes the recent advance in nanomaterials for electrochemical biosensors and bioassays. Biofunctionalization of nanomaterials for biosensors fabrication and their biomedical applications are discussed.

  14. Low-Cost Nanoribbon Sensors for Protein Analysis in Human Serum Using a Miniature Bead-Based Enzyme-Linked Immunosorbent Assay.

    PubMed

    Hu, Chunxiao; Zeimpekis, Ioannis; Sun, Kai; Anderson, Sally; Ashburn, Peter; Morgan, Hywel

    2016-05-01

    We describe a low cost thin-film transistor (TFT) nanoribbon sensor for detection of the inflammatory biomarker C-reactive protein (CRP) in human serum via a miniature bead-based enzyme-linked immunosorbent assay (ELISA). The TFT nanoribbon sensor measures the reaction products from the ELISA via pH changes. The bead-based ELISA decouples the protein functionalization steps from the sensor surface, increasing the signal and simplifying the assay. The ability to directly sense proteins in human serum in this way overcomes the Debye length limitation associated with nanowire and nanoribbon biosensors. Compared to classically fabricated nanowires, the TFT nanoribbon sensors are simple, extremely easy to fabricate, and should therefore be much cheaper to manufacture. TFT nanoribbon sensors, configured to measure pH, were used for quantitative detection of CRP spiked into human serum at concentrations as low as 0.2 ng/mL, which is 10 000 times lower than needed for diagnostic purposes, providing the potential for applications that require very high sensitivity. PMID:27035411

  15. A protein-protein binding assay using coated microtitre plates: increased throughput, reproducibility and speed compared to bead-based assays.

    PubMed

    Craig, Tim J; Ciufo, Leonora F; Morgan, Alan

    2004-07-30

    Protein-protein interactions, and the factors affecting them, are of fundamental importance to all biological systems. Surface plasmon resonance (SPR) and isothermal titration calorimetry (ITR) are powerful methods for assaying such interactions, but are expensive to implement. In contrast, bead-based pull-down assays using affinity tags such as glutathione-S-transferase (GST), require no specialist equipment. As a result, such assays are the most popular method for analysing protein-protein interactions, despite being time-consuming and prone to variability. In respect of these problems, we have modified this form of binding assay, using glutathione-coated 96-well plates rather than glutathione-Sepharose beads to bind the primary bait protein. Quantitation of bound protein utilises ELISA for purified proteins and scintillation counting for in vitro translated proteins, rather than the SDS-PAGE-based detection methods used in traditional bead-based assays. These modifications result in an approximately 10-fold increase in the number of samples that can be assayed daily, and allow results to be obtained within hours as opposed to days. We validate the modified assay by analysing the equilibrium binding of Munc18 and syntaxin, and also demonstrate that association and dissociation kinetics may be measured using this approach. The method we describe is generally applicable to any protein-protein interaction assay based on affinity tags and is amenable to automation, and so should benefit a wide range of biochemical research. PMID:15236910

  16. Early diagnosis of Irkut virus infection using magnetic bead-based serum peptide profiling by MALDI-TOF MS in a mouse model.

    PubMed

    Li, Nan; Liu, Ye; Hao, Zhuo; Zhang, Shoufeng; Hu, Rongliang; Li, Jiping

    2014-01-01

    Early diagnosis is important for the prompt post-exposure prophylaxis of lyssavirus infections. To diagnose Irkut virus (IRKV) infection during incubation in mice, a novel method using magnetic bead-based serum peptide profiling by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been established. For this test, serum peptides were concentrated by adsorption to and elution from the magnetic bead-based weak cation ion exchanger. Mass spectrograms obtained by MALDI-TOF MS were analyzed using ClinProTools bioinformatics software. Construction of the diagnostic model was performed using serum samples from mice infected with IRKV and rabies virus (RABV) BD06, Flury-LEP, and SRV9 (as controls). The method accurately diagnosed sera 2, 4 and 8 days after IRKV and RABV infections. The sensitivity, specificity, and total accuracy of diagnosis were 86.7%, 95.2%, and 92.9%, respectively. However, IRKV could not be differentiated from RABV 1 day after infection. The results of the present study indicate that serum peptide profiling by MALDI-TOF MS is a promising technique for the early clinical diagnosis of lyssavirus infections and needs to be further tested in humans and farm animals. PMID:24670473

  17. Bioassays Based on Molecular Nanomechanics

    DOE PAGESBeta

    Majumdar, Arun

    2002-01-01

    Recent experiments have shown that when specific biomolecular interactions are confined to one surface of a microcantilever beam, changes in intermolecular nanomechanical forces provide sufficient differential torque to bend the cantilever beam. This has been used to detect single base pair mismatches during DNA hybridization, as well as prostate specific antigen (PSA) at concentrations and conditions that are clinically relevant for prostate cancer diagnosis. Since cantilever motion originates from free energy change induced by specific biomolecular binding, this technique is now offering a common platform for label-free quantitative analysis of protein-protein binding, DNA hybridization DNA-protein interactions, and in general receptor-ligandmore » interactions. Current work is focused on developing “universal microarrays” of microcantilever beams for high-throughput multiplexed bioassays.« less

  18. In Vitro Biologic Activities of the Antimicrobials Triclocarban, Its Analogs, and Triclosan in Bioassay Screens: Receptor-Based Bioassay Screens

    PubMed Central

    Ahn, Ki Chang; Zhao, Bin; Chen, Jiangang; Cherednichenko, Gennady; Sanmarti, Enio; Denison, Michael S.; Lasley, Bill; Pessah, Isaac N.; Kültz, Dietmar; Chang, Daniel P.Y.; Gee, Shirley J.; Hammock, Bruce D.

    2008-01-01

    Background Concerns have been raised about the biological and toxicologic effects of the antimicrobials triclocarban (TCC) and triclosan (TCS) in personal care products. Few studies have evaluated their biological activities in mammalian cells to assess their potential for adverse effects. Objectives In this study, we assessed the activity of TCC, its analogs, and TCS in in vitro nuclear-receptor–responsive and calcium signaling bioassays. Materials and methods We determined the biological activities of the compounds in in vitro, cell-based, and nuclear-receptor–responsive bioassays for receptors for aryl hydrocarbon (AhR), estrogen (ER), androgen (AR), and ryanodine (RyR1). Results Some carbanilide compounds, including TCC (1–10 μM), enhanced estradiol (E2)-dependent or testosterone-dependent activation of ER- and AR-responsive gene expression up to 2.5-fold but exhibited little or no agonistic activity alone. Some carbanilides and TCS exhibited weak agonistic and/or antagonistic activity in the AhR-responsive bioassay. TCS exhibited antagonistic activity in both ER- and AR-responsive bioassays. TCS (0.1–10 μM) significantly enhanced the binding of [3H]ryanodine to RyR1 and caused elevation of resting cytosolic [Ca2+] in primary skeletal myotubes, but carbanilides had no effect. Conclusions Carbanilides, including TCC, enhanced hormone-dependent induction of ER- and AR-dependent gene expression but had little agonist activity, suggesting a new mechanism of action of endocrine-disrupting compounds. TCS, structurally similar to noncoplanar ortho-substituted poly-chlorinated biphenyls, exhibited weak AhR activity but interacted with RyR1 and stimulated Ca2+ mobilization. These observations have potential implications for human and animal health. Further investigations are needed into the biological and toxicologic effects of TCC, its analogs, and TCS. PMID:18795164

  19. Bioassay studies to determine OTEC's effect on phytoplankton activity

    SciTech Connect

    Carmiggelt, C.J.W.; Hartwig, E.O.; Commins, M.L.; Horne, A.J.

    1982-09-01

    The effect of artificially upwelled water (800m) on phytoplankton from 25m and 100m was simulated using five day bioassays. The results show that some enhancement of the phytoplankton populations in the receiving waters due to upwelling is likely to occur. The very small phytoplankton (< 5 um) are most important in this response. The magnitude of the biostimulation cannot be predicted from this study. Ammonia leaks, spills, and venting are probable in an operating OTEC plant. The bioassays show that additions of ammonia will produce biostimulaton only when the P/N ratio indicates nitrogen limitation. In the Hawaiian waters sampled N-limitation was not always present and varied with depth. No nitrogen fixation was detected. The magnitude of stimulation due to ammonia alone was generally less than the addition of upwelled water which is a more complete nutrient mixture.

  20. Comparison of three magnetic-bead-based RNA extraction methods for detection of cucumber green mottle mosaic virus by real-time RT-PCR.

    PubMed

    Zhao, Xiaoli; Zhou, Qi; Zhang, Lijie; Yan, Wenlong; Sun, Ning; Liang, Xinmiao; Deng, Congliang

    2015-07-01

    To determine the efficiency of RNA extraction methods based on magnetic beads, three different bead-based methods (one using silica-coated magnetic beads [SMNP], one using immunomagnetic beads conjugated to a specific antibody [IMB], and one using magnetic beads to nonspecifically adsorb virions [MNP]) were compared with the TRIzol method for the extraction of cucumber green mottle mosaic virus (CGMMV) RNA from cucumber leaves by real-time RT-PCR. The results indicated that the extraction efficiency of the SMNP method was 10 times higher than those of the IMB and MNP methods and 100 times higher than that of the TRIzol method. Therefore, the SMNP method could be considered for use in quarantine measures for the prevention and control of the disease caused by CGMMV. PMID:25951973

  1. Isolation of prostate cancer cell subpopulations of functional interest by use of an on-chip magnetic bead-based cell separator

    NASA Astrophysics Data System (ADS)

    Estes, Matthew D.; Ouyang, Bin; Ho, Shuk-mei; Ahn, Chong H.

    2009-09-01

    This work presents the design, fabrication and characterization of a modular magnetic bead-based cell separation device developed for the sequential sorting of a heterogeneous prostate cancer (CaP) cell population. The chief aim is cell sorting carried out on the basis of surface marker expression, serially selecting cellular subpopulations for capture by the use of antibody-coated magnetic beads. The markers of interest, prostate specific membrane antigen (PSMA) and CD10 were selected for their relevance to ongoing CaP development research. The separation device was fabricated out of plastic, by the use of cyclic olefin copolymer (COC) injection molding, nickel-iron electroplating and thermoplastic fusion bonding. Effective depletion and enrichment of cell subsets based on multiple surface markers was achieved. Various flow rates and incubation times were tested for optimizing the sorting procedure.

  2. A flexible multiplex bead-based assay for detecting germline CDKN2A and CDK4 variants in melanoma-prone kindreds.

    PubMed

    Lang, Julie M; Shennan, Michael; Njauw, Jenny C-N; Luo, Su; Bishop, Julia N; Harland, Mark; Hayward, Nicholas K; Tucker, Margaret A; Goldstein, Alisa M; Landi, Maria T; Puig, Susana; Gruis, Nelleke A; Bergman, Wilma; Bianchi-Scarra, Giovanna; Ghiorzo, Paola; Hogg, David; Tsao, Hensin

    2011-02-01

    The presence of recurrent high-risk mutations in cyclin-dependent kinase inhibitor 2A/cyclin-dependent kinase 4 (CDKN2A/CDK4) among melanoma-prone families suggests that a high-throughput, multiplex assay could serve as an effective initial screening tool. To this end, we have developed a multiplex bead-based assay for high-throughput CDKN2A/CDK4 genotyping in the context of familial melanoma. Genomic DNA from 1,603 subjects (1,005 in training set and 598 in validation set) were amplified by multiplex PCR using five CDKN2A/CDK4 primer sets followed by multiplex allele-specific primer extension for 39 distinct germline variants. The products were then sorted and analyzed using the Luminex xMAP system. Genotypes were compared with previously determined sequence data. In the Toronto training cohort, all 145 samples with known variants were detected by the bead assay (100% concordance). Analysis of the 598 samples from the GenoMEL validation set led to identification of 150/155 expected variants (96.77%). Overall, the bead assay correctly genotyped 1,540/1,603 (96.07%) of all individuals in the study and 1,540/1,545 (99.68%) of individuals whose variants were represented in the probe set. Out of a total of 62,517 allelic calls, 62,512 (99.99%) were correctly assigned. The multiplex bead-based assay is an accurate method for genotyping CDKN2A/CDK4 variants and is potentially useful in genotyping low-to-moderate melanoma risk single-nucleotide polymorphisms. PMID:21085193

  3. A bioaccumulation bioassay for freshwater sediments

    USGS Publications Warehouse

    Mac, Michael J.; Noguchi, George E.; Hesselberg, Robert J.; Edsall, Carol C.; Shoesmith, John A.; Bowker, James D.

    1990-01-01

    A laboratory bioassay is described for determining the bioavailability of contaminants from freshwater sediments. The bioassay consists of 10-d exposures to whole sediments under flow-through conditions. After testing five species, the fathead minnow (Pimephales promelas) and the earthworm (Lubricus terrestris) were recommended for use in the test. When the availability of polychlorinated biphenyls (PCBs), Hg and Zn from Great Lakes sediments was examined in laboratory exposures, only the PCBs were accumulated. A field validation study demonstrated that the magnitude of accumulation in laboratory exposures was similar to that in organisms caged in the field. A protocol is recommended for using the test as a standardized bioaccumulation bioassay.

  4. Two-generation saccharin bioassays.

    PubMed

    Arnold, D L

    1983-04-01

    The controversy regarding the safety of saccharin for human consumption started shortly after its discovery over 100 years ago and has yet to subside appreciably. The consumption of saccharin, particularly in North America, began to escalate when the U.S. Food and Drug Administration set new standards of identity which allowed foods containing artificial sweeteners to be promoted as "nonnutritive" or "noncaloric" sweeteners for use by the general public. In 1969, when cyclamates were banned, at least 10 single-generation feeding studies were undertaken with saccharin to more accurately assess the potential toxicological consequences resulting from the anticipated increase in its consumption. None of these studies resulted in any overt regulatory action. Subsequently, the introduction of the two-generation chronic toxicity/carcinogenicity bioassay added a new tool to the toxicologist's arsenal. Three two-generation studies using saccharin have since been conducted. The results from these studies clearly show that when rats were exposed to diets containing 5 or 7.5% sodium saccharin from the time of conception to death, an increased frequency of urinary bladder cancers was found, predominantly in the males. While some study results suggested that impurities in commercial saccharin or the presence of urinary tract calculi may have been responsible for the observed bladder tumors, it now appears that these possibilities are highly unlikely. The mechanism by which saccharin elicited the bladder tumors using the two-generation experiment has not been ascertained. PMID:6347682

  5. Bioassays for Monitoring Insecticide Resistance

    PubMed Central

    Miller, Audra L.E.; Tindall, Kelly; Leonard, B. Rogers

    2010-01-01

    Pest resistance to pesticides is an increasing problem because pesticides are an integral part of high-yielding production agriculture. When few products are labeled for an individual pest within a particular crop system, chemical control options are limited. Therefore, the same product(s) are used repeatedly and continual selection pressure is placed on the target pest. There are both financial and environmental costs associated with the development of resistant populations. The cost of pesticide resistance has been estimated at approximately $ 1.5 billion annually in the United States. This paper will describe protocols, currently used to monitor arthropod (specifically insects) populations for the development of resistance. The adult vial test is used to measure the toxicity to contact insecticides and a modification of this test is used for plant-systemic insecticides. In these bioassays, insects are exposed to technical grade insecticide and responses (mortality) recorded at a specific post-exposure interval. The mortality data are subjected to Log Dose probit analysis to generate estimates of a lethal concentration that provides mortality to 50% (LC50) of the target populations and a series of confidence limits (CL's) as estimates of data variability. When these data are collected for a range of insecticide-susceptible populations, the LC50 can be used as baseline data for future monitoring purposes. After populations have been exposed to products, the results can be compared to a previously determined LC50 using the same methodology. PMID:21248689

  6. PHOXOCEPHALID AMPHIPOD BIOASSAY FOR MARINE SEDIMENT TOXICITY

    EPA Science Inventory

    The relative toxicity of marine sediment can be accurately determined through acute, static bioassays with the phoxocepalid amphipod Repoxynius abronius. Mortality and sublethal effects on emergence from sediment and reburial behavior are determined after ten day exposure in 1-L ...

  7. Bioassay criteria for environmental restoration workers

    SciTech Connect

    Carbaugh, E.H.; Bihl, D.E.

    1993-01-01

    Environmental restoration (ER) work at the U. S. Department of Energy Hanford Site posed questions concerning when to perform bioassay monitoring of workers for potential intakes of radioactivity. Application of criteria originally developed for use inside radionuclide processing facilities to ER work resulted in overly restrictive bioassay requirements. ER work typically involves site characterization or, excavating large quantities of potentially contaminated soil, rather than working with concentrated quantities of radioactivity as in a processing facility. An improved approach, tailored to ER work, provided soil contamination concentrations above which worker bioassay would be required. Soil concentrations were derived assuming acute or chronic intakes of 2% of an Annual Limit on Intake (ALI), or a potential committed effective dose equivalent of 100 mrem, and conservative dust loading of air from the work. When planning ER work, the anticipated soil concentration and corresponding need for bioassay could be estimated from work-site historical records. Once site work commenced, soil sampling and work-place surveys could be used to determine bioassay needs. This approach substantially reduced the required number of bioassay samples with corresponding reductions in analytical costs, schedules, and more flexible work-force management. (Work supported by the US Department of Energy under contract DOE-AC06-76RLO 1830.)

  8. A Colorimetric Bioassay for Perchlorate

    NASA Astrophysics Data System (ADS)

    Heinnickel, M. L.; Smith, S.; Coates, J. D.

    2007-12-01

    Recognition of perchlorate (ClO4-) as a widespread contaminant across the United States and its potential adverse affects towards human health has motivated the EPA to place ClO4- on its contaminant candidate list for drinking water supplies. While a federal MCL has not yet been set, a recommended public health goal of 1 ppb (μg.L-1) was established by the US EPA in 2002. To date, methods of detection require use of sensitive ion chromatographic equipment that are expensive, time consuming, and require highly trained personnel for use. Our studies are focused on the development of a highly sensitive, simple, and robust colorimetric bioassay based on the primary enzyme involved in microbial ClO4- reduction, the perchlorate reductase (Pcr). A previously published assay used reduced methyl viologen (MV, the dye is reduced with sodium hydrosulfite) as an electron donor to demonstrate Pcr activity. The assay directly correlates the amount of MV oxidized with the amount of ClO4- reduced by assuming a transfer of four electrons. To test this assumption, we compared actual concentrations of MV oxidized to ClO4- reduced in this assay. ClO4- concentrations were determined using a Dionex ICS-500 ion chromatography system, while MV concentrations were determined using a standard curve generated at 578 nm. Comparisons between the two revealed that twelve molecules of MV were oxidized for each molecule of ClO4- reduced. The oxidation of these additional eight MV molecules is explained by the interaction of the dye with chlorite (the product of the Pcr reaction) and other contaminants that could be present in the enzyme prep. This unsettling result indicated this assay would be problematic for the detection of ClO4- in soil, which has many chemicals that could react with MV. To improve upon this assay, we have tried to reduce ClO4- using less reactive dyes and reductants. The reductants ascorbic acid, NADH, and dithiothreitol drive Pcr catalyzed ClO4- reduction, however, they

  9. The Limits of Two-Year Bioassay Exposure Regimens for Identifying Chemical Carcinogens

    PubMed Central

    Huff, James; Jacobson, Michael F.; Davis, Devra Lee

    2008-01-01

    Background Chemical carcinogenesis bioassays in animals have long been recognized and accepted as valid predictors of potential cancer hazards to humans. Most rodent bioassays begin several weeks after birth and expose animals to chemicals or other substances, including workplace and environmental pollutants, for 2 years. New findings indicate the need to extend the timing and duration of exposures used in the rodent bioassay. Objectives In this Commentary, we propose that the sensitivity of chemical carcinogenesis bio-assays would be enhanced by exposing rodents beginning in utero and continuing for 30 months (130 weeks) or until their natural deaths at up to about 3 years. Discussion Studies of three chemicals of different structures and uses—aspartame, cadmium, and toluene—suggest that exposing experimental animals in utero and continuing exposure for 30 months or until their natural deaths increase the sensitivity of bioassays, avoid false-negative results, and strengthen the value and validity of results for regulatory agencies. Conclusions Government agencies, drug companies, and the chemical industry should conduct and compare the results of 2-year bioassays of known carcinogens or chemicals for which there is equivocal evidence of carcinogenicity with longer-term studies, with and without in utero exposure. If studies longer than 2 years and/or with in utero exposure are found to better identify potential human carcinogens, then regulatory agencies should promptly revise their testing guidelines, which were established in the 1960s and early 1970s. Changing the timing and dosing of the animal bioassay would enhance protection of workers and consumers who are exposed to potentially dangerous workplace or home contaminants, pollutants, drugs, food additives, and other chemicals throughout their lives. PMID:19057693

  10. Facile synthesis of magnetic-/pH-responsive hydrogel beads based on Fe3O4 nanoparticles and chitosan hydrogel as MTX carriers for controlled drug release.

    PubMed

    Wu, Juan; Jiang, Wei; Tian, Renbing; Shen, Yewen; Jiang, Wei

    2016-10-01

    In the present study, methotrexate (MTX)-encapsulated magnetic-/pH-responsive hydrogel beads based on Fe3O4 nanoparticles and chitosan were successfully prepared through a one-step gelation process, which is a very facile, economic and environmentally friendly route. The developed hydrogel beads exhibited homogeneous porous structure and super-paramagnetic responsibility. MTX can be successfully encapsulated into magnetic chitosan hydrogel beads, and the drug encapsulation efficiency (%) and encapsulation content (%) were 93.8 and 6.28%, respectively. In addition, the drug release studies in vitro indicated that the MTX-encapsulated magnetic chitosan hydrogel beads had excellent pH-sensitivity, 90.6% MTX was released from the magnetic chitosan hydrogel beads within 48 h at pH 4.0. WST-1 assays in human liver hepatocellular carcinoma cells (HepG2) demonstrated that the MTX-encapsulated magnetic chitosan hydrogel beads had good cytocompatibility and high anti-tumor activity. Therefore, our results revealed that the MTX-encapsulated magnetic chitosan hydrogel beads would be a competitive candidate for controlled drug release in the area of targeted cancer therapy in the near future. PMID:27464586

  11. Liquid carry-over in an injection moulded all-polymer chip system for immiscible phase magnetic bead-based solid-phase extraction

    NASA Astrophysics Data System (ADS)

    Kistrup, Kasper; Skotte Sørensen, Karen; Wolff, Anders; Fougt Hansen, Mikkel

    2015-04-01

    We present an all-polymer, single-use microfluidic chip system produced by injection moulding and bonded by ultrasonic welding. Both techniques are compatible with low-cost industrial mass-production. The chip is produced for magnetic bead-based solid-phase extraction facilitated by immiscible phase filtration and features passive liquid filling and magnetic bead manipulation using an external magnet. In this work, we determine the system compatibility with various surfactants. Moreover, we quantify the volume of liquid co-transported with magnetic bead clusters from Milli-Q water or a lysis-binding buffer for nucleic acid extraction (0.1 (v/v)% Triton X-100 in 5 M guanidine hydrochloride). A linear relationship was found between the liquid carry-over and mass of magnetic beads used. Interestingly, similar average carry-overs of 1.74(8) nL/μg and 1.72(14) nL/μg were found for Milli-Q water and lysis-binding buffer, respectively.

  12. Acarine attractants: Chemoreception, bioassay, chemistry and control.

    PubMed

    Carr, Ann L; Roe, Michael

    2016-07-01

    The Acari are of significant economic importance in crop production and human and animal health. Acaricides are essential for the control of these pests, but at the same time, the number of available pesticides is limited, especially for applications in animal production. The Acari consist of two major groups, the mites that demonstrate a wide variety of life strategies, i.e., herbivory, predation and ectoparasitism, and ticks which have evolved obligatory hematophagy. The major sites of chemoreception in the acarines are the chelicerae, palps and tarsi on the forelegs. A unifying name, the "foretarsal sensory organ" (FSO), is proposed for the first time in this review for the sensory site on the forelegs of all acarines. The FSO has multiple sensory functions including olfaction, gustation, and heat detection. Preliminary transcriptomic data in ticks suggest that chemoreception in the FSO is achieved by a different mechanism from insects. There are a variety of laboratory and field bioassay methods that have been developed for the identification and characterization of attractants but minimal techniques for electrophysiology studies. Over the past three to four decades, significant progress has been made in the chemistry and analysis of function for acarine attractants in mites and ticks. In mites, attractants include aggregation, immature female, female sex and alarm pheromones; in ticks, the attraction-aggregation-attachment, assembly and sex pheromones; in mites and ticks host kairomones and plant allomones; and in mites, fungal allomones. There are still large gaps in our knowledge of chemical communication in the acarines compared to insects, especially relative to acarine pheromones, and more so for mites than ticks. However, the use of lure-and-kill and lure-enhanced biocontrol strategies has been investigated for tick and mite control, respectively, with significant environmental advantages which warrant further study. PMID:27265828

  13. Poultry litter toxicity comparison from various bioassays

    SciTech Connect

    Gupta, G.; Kelly, P. )

    1992-01-01

    Poultry litter contains many toxic chemicals including Cu, As, Pb, Cd, Hg, Se and PCBs. Poultry litter leachate has been shown to be more toxic to marine luminescent organisms (Photobacterium phosphoreum) than other farm animal manures. A comparison of toxicity of the poultry litter leachate was undertaken using various bioassays. The EC{sub 50} (or LC{sub 50}) value for the leachate with the Microtox and Daphnia bioassays was 2.9 g/L/ Nitrobacter and Pseudomonas bioassays were not useful in determining the leachate toxicity because of the nutritional properties of the litter. Poultry litter leachate was found to be mutagenic to strains TA 97, TA 98, TA 100 and TA 102 using the Ames Test.

  14. RECOMMENDATIONS FOR UO3 PLANT BIOASSAY

    SciTech Connect

    Carbaugh, Eugene H.

    2010-07-12

    Alternative urine bioassay programs are described for application with decontamination and decommissioning activities at the Hanford UO3 Plant. The alternatives are based on quarterly or monthly urine bioassay for recycled uranium, assuming multiple acute inhalation intakes of recycled uranium occurring over a year. The inhalations are assumed to be 5µm AMAD particles of 80% absorption type F and 20% absorption type M. Screening levels, expressed as daily uranium mass excretion rates in urine, and the actions associated with these levels are provided for both quarterly and monthly sampling frequencies.

  15. Experiences with Non-traditional Bioassay Methods in a Plutonium Processing Line

    SciTech Connect

    La Bone, T.R.

    2003-10-17

    An incident in an Savannah River Site (SRS) plutonium processing line (FB-Line) in 1999 highlighted the fact insoluble forms of plutonium exist at SRS that may not be readily monitored with the routine bioassay programs traditionally used at this site. To address this issue, a study was conducted in FB-Line with 21 participants for a year ending in July 2002. The purpose of the study was to examine the use of three non-traditional monitoring methods and, based on this experience, recommend a routine bioassay program that is capable of monitoring workers potentially exposed to insoluble plutonium. These non-traditional monitoring methods are personal air sampling (PAS), thermal ionization mass spectrometry (TIMS) of urine samples, and routine fecal bioassay. The main conclusions and recommendations of the study are: (1) A routine TIMS urine bioassay program, which is called the enhanced bioassay program (EBP), is recommended for workers in SRS facilities that have a reasonable potential for exposure to insoluble forms of plutonium. (2) Under certain conditions the EBP could result in onerous work restrictions. A contingency plan involving the use of PAS is recommended in this case. PAS is also recommended for workers who have had historic intakes of plutonium that interfere with the detection and interpretation of future intakes of insoluble plutonium. (3) For the EBP to be successful it must be used only for those workers who have a reasonable potential for exposure to insoluble plutonium, and these workers must take all necessary precautions to avoid cross-contamination of the urine (and follow-up fecal) samples. (4) Fecal bioassay is an important tool for follow-up to abnormal events, but routine fecal bioassay is not recommended. (5) The PAS data clearly shows that workers are exposed to low levels of airborne plutonium, but the participants appear to be unlikely to exceed a committed effective dose equivalent of 100 mrem from these exposures.

  16. BIOASSAY-DIRECTED CHEMICAL ANALYSIS IN ENVIRONMENTAL RESEARCH

    EPA Science Inventory

    The use of short-term bioassay tests in conjunction with analytical measurements, constitute a powerful tool for identifying important environmental contaminants. The authors have coined the terminology 'bioassay directed chemical analysis' to best describe this marriage of analy...

  17. Adoptive transfer of pTRP2-specific CTLs expanding by bead-based artificial antigen-presenting cells mediates anti-melanoma response.

    PubMed

    Lu, Xiaoling; Jiang, Xiaobing; Liu, Ruen; Zhao, Hongyang; Liang, Zhihui

    2008-11-18

    Cytotoxic CD8(+) T cells are key effectors in the immunotherapy of malignant and viral diseases. However, the lack of efficient methods for their in vitro priming and expansion has become a bottleneck to the development of vaccines and adoptive transfer strategies. Synthetic artificial antigen-presenting cells (aAPCs) are now emerging as an attractive tool for eliciting and expanding CTL responses. This study reported a novel approach for targeting malignant melanoma with pTRP2-specific cytotoxic T lymphocytes (CTLs) expanded from the C57BL/6 splenocytes by multiple stimulations with aAPCs made by coating H-2K(b)-Ig/pTRP2 dimeric complexes, anti-CD28 antibody, 4-1BBL molecules and CD83 molecules to cell-sized latex beads. The induced CTLs exhibited specific lysis against RMA-S cells pulsed with the peptide pTRP2 and H-2K(b+) melanoma cells expressing TRP2, while a murine Lewis lung carcinoma cell line 3LL could not be recognized by the CTLs. The peptide-specific activity was inhibited by anti-H-2K(b) monoclonal antibody Y3. Adoptive Transfer of CTLs specific for malignant melanoma expanding by the aAPCs can mediate effective anti-melanoma response. These results suggested the bead-based aAPCs coated with an MHC-Ig/peptide complex, anti-CD28 antibody, 4-1BBL and CD83 could provide a useful tool for the reproducible expansion of specific CTLs for adoptive immunotherapy. PMID:18621475

  18. Semiconductor sensor embedded microfluidic chip for protein biomarker detection using a bead-based immunoassay combined with deoxyribonucleic acid strand labeling.

    PubMed

    Lin, Yen-Heng; Peng, Po-Yu

    2015-04-15

    Two major issues need to be addressed in applying semiconductor biosensors to detecting proteins in immunoassays. First, the length of the antibody on the sensor surface surpasses the Debye lengths (approximately 1 nm, in normal ionic strength solution), preventing certain specifically bound proteins from being tightly attached to the sensor surface. Therefore, these proteins do not contribute to the sensor's surface potential change. Second, these proteins carry a small charge and can be easily affected by the pH of the surrounding solution. This study proposes a magnetic bead-based immunoassay using a secondary antibody to label negatively charged DNA fragments for signal amplification. An externally imposed magnetic force attaches the analyte tightly to the sensor surface, thereby effectively solving the problem of the analyte protein's distance to the sensor surface surpassing the Debye lengths. In addition, a normal ion intensity buffer can be used without dilution for the proposed method. Experiments revealed that the sensitivity can be improved by using a longer DNA fragment for labeling and smaller magnetic beads as solid support for the antibody. By using a 90 base pair DNA label, the signal was 15 times greater than that without labeling. In addition, by using a 120 nm magnetic bead, a minimum detection limit of 12.5 ng mL(-1) apolipoprotein A1 can be measured. Furthermore, this study integrates a semiconductor sensor with a microfluidic chip. With the help of microvalves and micromixers in the chip, the length of the mixing step for each immunoassay has been reduced from 1h to 20 min, and the sample volume has been reduced from 80 μL to 10 μL. In practice, a protein biomarker in a urinary bladder cancer patient's urine was successfully measured using this technique. This study provides a convenient and effective method to measure protein using a semiconductor sensor. PMID:25818137

  19. Micro-organism distribution sampling for bioassays

    NASA Technical Reports Server (NTRS)

    Nelson, B. A.

    1975-01-01

    Purpose of sampling distribution is to characterize sample-to-sample variation so statistical tests may be applied, to estimate error due to sampling (confidence limits) and to evaluate observed differences between samples. Distribution could be used for bioassays taken in hospitals, breweries, food-processing plants, and pharmaceutical plants.

  20. EDC BIOASSAYS FOR RISK MANAGEMENT PROJECTS

    EPA Science Inventory

    Overall goal for this research is to develop 3 bioassays for use in EDC projects across NRMRL (estrogenic, androgenic and thyroid assays). Currently, research is focused on estrogenic assays. A literature search was conducted to identify potential assays. The Yeast Estrogen Sc...

  1. Brine Shrimp Bioassays: A Useful Technique in Biological Investigations

    ERIC Educational Resources Information Center

    Rice, Stanley A.; Maness, Ian B.

    2004-01-01

    A technique to measure the potency of leaf compounds against herbivores with the use of a bioassay is described. Bioassays are useful in classes where students have career plans like medicine in which bioassays can be used as tools for screening plants for possible medicinal potency.

  2. BIOASSAY-DIRECTED FRACTIONATION OF ORGANIC CONTAMINANTS IN AN ESTUARINE SEDIMENT USING THE NEW MUTAGENIC BIOASSAY, MUTATOX

    EPA Science Inventory

    Bioassay-directed fractionation of organic compounds was performed on an organic solvent extract of a contaminated estuarine sediment from Black Rock Harbor, Connecticut, using the new mutagenic bioassay, Mutatox-. hemical fractionation methods of the sediment extract included si...

  3. Bioassaying for ozone with pollen systems

    SciTech Connect

    Feder, W.A.

    1981-01-01

    Sensitivity to ozone of pollen germinating in vitro is closely correlated with ozone sensitivity of the pollen parent. Ozone-sensitive and tolerant pollen populations have been identified in tobacco, petunia, and tomato cultivars. The rate of tube elongation can be reversibly slowed or stopped by exposure to low concentrations of ozone. The performance of selected pollen populations can then be used to bioassay ozone in ambient air by introducing the air sample into a growth chamber where ozone-sensitive pollen in growing. Year-round pollen producion can be achieved in the greenhouse. Harvested pollen can be tested, packaged, and transported to user facilities without loss of vigor. Pollen populations are inexpensive to produce, respond reliably, and are simple to use as a bioassay for air quality.

  4. Perspectives in avoidance-preference bioassays

    SciTech Connect

    Steele, C.W.; Taylor, D.H.; Strickler-Shaw, S.

    1996-12-31

    Although behavioral endpoints are used in hazard assessment, establishment of water quality criteria and assessment of a contaminant`s hazard to aquatic life rely primarily on standard acute and chronic toxicity tests. Sublethal effects of pollutants should, however, be of major concern because more organisms experience sublethal rather than acutely or chronically lethal exposures of contaminants. The avoidance-preference approach to behavioral bioassays is very useful in screening pollutants for which the mechanisms of perception or response are largely unknown. The underlying philosophy of these studies is that an animal which perceives a chemical can be attracted or repulsed by it. No response is frequently assumed to indicate lack of perception. All three responses have broad ecological implications. The authors discuss the conditions required for performing avoidance-preference bioassays, as well as their sensitivities, advantages, and limitations. In this regard, a comparative approach is used in examining the results of avoidance-preference bioassays with zebrafish in two different apparatuses. Finally, they compare the results of avoidance-preference studies with other measures of the behavioral toxicity of lead to tadpoles.

  5. A versatile electrowetting-based digital microfluidic platform for quantitative homogeneous and heterogeneous bio-assays

    NASA Astrophysics Data System (ADS)

    Vergauwe, Nicolas; Witters, Daan; Ceyssens, Frederik; Vermeir, Steven; Verbruggen, Bert; Puers, Robert; Lammertyn, Jeroen

    2011-05-01

    Electrowetting-on-dielectric (EWOD) lab-on-a-chip systems have already proven their potential within a broad range of bio-assays. Nevertheless, research on the analytical performance of those systems is limited, yet crucial for a further breakthrough in the diagnostic field. Therefore, this paper presents the intrinsic possibilities of an EWOD lab-on-a-chip as a versatile platform for homogeneous and heterogeneous bio-assays with high analytical performance. Both droplet dispensing and splitting cause variations in droplet size, thereby directly influencing the assay's performance. The extent to which they influence the performance is assessed by a theoretical sensitivity analysis, which allows the definition of a basic framework for the reduction of droplet size variability. Taking advantage of the optimized droplet manipulations, both homogeneous and heterogeneous bio-assays are implemented in the EWOD lab-on-a-chip to demonstrate the analytical capabilities and versatility of the device. A fully on-chip enzymatic assay is realized with high analytical performance. It demonstrates the promising capabilities of an EWOD lab-on-a-chip in food-related and medical applications, such as nutritional and blood analyses. Further, a magnetic bio-assay for IgE detection using superparamagnetic nanoparticles is presented whereby the nanoparticles are used as solid carriers during the bio-assay. Crucial elements are the precise manipulation of the superparamagnetic nanoparticles with respect to dispensing and separation. Although the principle of using nano-carriers is demonstrated for protein detection, it can be easily extended to a broader range of bio-related applications like DNA sensing. In heterogeneous bio-assays the chip surface is actively involved during the execution of the bio-assay. Through immobilization of specific biological compounds like DNA, proteins and cells a reactive chip surface is realized, which enhances the bio-assay performance. To demonstrate

  6. An in vitro rainbow trout cell bioassay for aryl hydrocarbon receptor-mediated toxins

    SciTech Connect

    Richter, C.A.; Tieber, V.L.; Giesy, J.P.; Denison, M.S.

    1997-03-01

    Halogenated aromatic hydrocarbons (HAHs) and other chemicals that act as aryl hydrocarbon (Ah) receptor (AhR) agonists cause a variety of toxicity effects. In sac fry of many fish species, these effects include blue-sac disease and mortality. Because HAHs occur in complex mixtures, their toxicity in the environment is difficult to predict. A bioassay useful in predicting AhR-mediated toxicity to fish was developed using the RTH-149 rainbow trout hepatoma cell line. Stable transfection of this cell line with the pGudLuc 1.1 plasmid, which contains a firefly luciferase reporter gene under the transcriptional regulation of dioxin responsive enhancers, has produced a recombinant cell line designated Remodulated Lightning Trout (RLT 2.0). The RLT 2.0 bioassay method detection limit for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is 4 pM. The responses of the RLT 2.0 bioassay to TCDD and several HAH congeners closely matched the responses observed in vivo in fish. The RLT 2.0 bioassay can provide an integrative measure of the total AhR-mediated toxic activity of complex mixtures to fish. The assay will be useful in screening environmental extracts, guiding chemical analysis, and interpreting the AhR-mediated mechanism of toxicity.

  7. In-situ bioassays using caged bivalves

    SciTech Connect

    Salazar, M.H.; Salazar, S.M.

    1995-12-31

    It is important to make the distinction between chemical measurements to assess bioaccumulation potential versus biological measurements to assess potential bioeffects because bioaccumulation is not a bioeffect. Caging provides a unique opportunity to make synoptic measurements of each and facilitates making these measurements over space and time. Measuring bioaccumulation in resident and transplanted bivalves has probably been the most frequently used form of an in-situ bioassay because bivalves concentrate chemicals in their tissues. They are also easy to collect, cage, and measure. The authors have refined bivalve bioassay methods by minimizing the size range of test animals, making repetitive measurements of the same individuals, and standardizing test protocols for a variety of applications. They are now attempting to standardize criteria for accepting and interpreting data in the same way that laboratory bioassays have been standardized. Growth measurements can serve two purposes in this assessment strategy: (1) An integrated biological response endpoint that is easily quantifiable and with significance to the population, and (2) A means of calibrating bioaccumulation by assessing the relative health and physiological state of tissues that have accumulated the chemicals. In general, the authors have found the highest bioconcentration factors associated with the highest growth rates, the highest concentrations ({micro}g/g) of chemicals in juvenile mussels, and the highest chemical content ({micro}g/animal) in adult mussels. Without accounting for possible dilution of chemical concentrations by tissue growth or magnification through degrowth, contaminant concentrations can be misleading. Examples are provided for the Sudbury River in Massachusetts (Elliptio complanata), San Diego Bay (Mytilus galloprovincialis), and the Harbor Island Superfund Site in Puget Sound (Mytilus trossulus).

  8. A Multichannel Bioluminescence Determination Platform for Bioassays.

    PubMed

    Kim, Sung-Bae; Naganawa, Ryuichi

    2016-01-01

    The present protocol introduces a multichannel bioluminescence determination platform allowing a high sample throughput determination of weak bioluminescence with reduced standard deviations. The platform is designed to carry a multichannel conveyer, an optical filter, and a mirror cap. The platform enables us to near-simultaneously determine ligands in multiple samples without the replacement of the sample tubes. Furthermore, the optical filters beneath the multichannel conveyer are designed to easily discriminate colors during assays. This optical system provides excellent time- and labor-efficiency to users during bioassays. PMID:27424912

  9. Evolving BioAssay Ontology (BAO): modularization, integration and applications

    PubMed Central

    2014-01-01

    The lack of established standards to describe and annotate biological assays and screening outcomes in the domain of drug and chemical probe discovery is a severe limitation to utilize public and proprietary drug screening data to their maximum potential. We have created the BioAssay Ontology (BAO) project (http://bioassayontology.org) to develop common reference metadata terms and definitions required for describing relevant information of low-and high-throughput drug and probe screening assays and results. The main objectives of BAO are to enable effective integration, aggregation, retrieval, and analyses of drug screening data. Since we first released BAO on the BioPortal in 2010 we have considerably expanded and enhanced BAO and we have applied the ontology in several internal and external collaborative projects, for example the BioAssay Research Database (BARD). We describe the evolution of BAO with a design that enables modeling complex assays including profile and panel assays such as those in the Library of Integrated Network-based Cellular Signatures (LINCS). One of the critical questions in evolving BAO is the following: how can we provide a way to efficiently reuse and share among various research projects specific parts of our ontologies without violating the integrity of the ontology and without creating redundancies. This paper provides a comprehensive answer to this question with a description of a methodology for ontology modularization using a layered architecture. Our modularization approach defines several distinct BAO components and separates internal from external modules and domain-level from structural components. This approach facilitates the generation/extraction of derived ontologies (or perspectives) that can suit particular use cases or software applications. We describe the evolution of BAO related to its formal structures, engineering approaches, and content to enable modeling of complex assays and integration with other ontologies and

  10. Evolving BioAssay Ontology (BAO): modularization, integration and applications.

    PubMed

    Abeyruwan, Saminda; Vempati, Uma D; Küçük-McGinty, Hande; Visser, Ubbo; Koleti, Amar; Mir, Ahsan; Sakurai, Kunie; Chung, Caty; Bittker, Joshua A; Clemons, Paul A; Brudz, Steve; Siripala, Anosha; Morales, Arturo J; Romacker, Martin; Twomey, David; Bureeva, Svetlana; Lemmon, Vance; Schürer, Stephan C

    2014-01-01

    The lack of established standards to describe and annotate biological assays and screening outcomes in the domain of drug and chemical probe discovery is a severe limitation to utilize public and proprietary drug screening data to their maximum potential. We have created the BioAssay Ontology (BAO) project (http://bioassayontology.org) to develop common reference metadata terms and definitions required for describing relevant information of low-and high-throughput drug and probe screening assays and results. The main objectives of BAO are to enable effective integration, aggregation, retrieval, and analyses of drug screening data. Since we first released BAO on the BioPortal in 2010 we have considerably expanded and enhanced BAO and we have applied the ontology in several internal and external collaborative projects, for example the BioAssay Research Database (BARD). We describe the evolution of BAO with a design that enables modeling complex assays including profile and panel assays such as those in the Library of Integrated Network-based Cellular Signatures (LINCS). One of the critical questions in evolving BAO is the following: how can we provide a way to efficiently reuse and share among various research projects specific parts of our ontologies without violating the integrity of the ontology and without creating redundancies. This paper provides a comprehensive answer to this question with a description of a methodology for ontology modularization using a layered architecture. Our modularization approach defines several distinct BAO components and separates internal from external modules and domain-level from structural components. This approach facilitates the generation/extraction of derived ontologies (or perspectives) that can suit particular use cases or software applications. We describe the evolution of BAO related to its formal structures, engineering approaches, and content to enable modeling of complex assays and integration with other ontologies and

  11. Urine sample collection protocols for bioassay samples

    SciTech Connect

    MacLellan, J.A.; McFadden, K.M.

    1992-11-01

    In vitro radiobioassay analyses are used to measure the amount of radioactive material excreted by personnel exposed to the potential intake of radioactive material. The analytical results are then used with various metabolic models to estimate the amount of radioactive material in the subject`s body and the original intake of radioactive material. Proper application of these metabolic models requires knowledge of the excretion period. It is normal practice to design the bioassay program based on a 24-hour excretion sample. The Hanford bioassay program simulates a total 24-hour urine excretion sample with urine collection periods lasting from one-half hour before retiring to one-half hour after rising on two consecutive days. Urine passed during the specified periods is collected in three 1-L bottles. Because the daily excretion volume given in Publication 23 of the International Commission on Radiological Protection (ICRP 1975, p. 354) for Reference Man is 1.4 L, it was proposed to use only two 1-L bottles as a cost-saving measure. This raised the broader question of what should be the design capacity of a 24-hour urine sample kit.

  12. Urine sample collection protocols for bioassay samples

    SciTech Connect

    MacLellan, J.A.; McFadden, K.M.

    1992-11-01

    In vitro radiobioassay analyses are used to measure the amount of radioactive material excreted by personnel exposed to the potential intake of radioactive material. The analytical results are then used with various metabolic models to estimate the amount of radioactive material in the subject's body and the original intake of radioactive material. Proper application of these metabolic models requires knowledge of the excretion period. It is normal practice to design the bioassay program based on a 24-hour excretion sample. The Hanford bioassay program simulates a total 24-hour urine excretion sample with urine collection periods lasting from one-half hour before retiring to one-half hour after rising on two consecutive days. Urine passed during the specified periods is collected in three 1-L bottles. Because the daily excretion volume given in Publication 23 of the International Commission on Radiological Protection (ICRP 1975, p. 354) for Reference Man is 1.4 L, it was proposed to use only two 1-L bottles as a cost-saving measure. This raised the broader question of what should be the design capacity of a 24-hour urine sample kit.

  13. Aspirator Gun for High-Throughput Mosquito Bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We describe an innovative aspirator gun designed to transfer anaesthetized mosquitoes directly into glass bioassay tubes. The gun has been used for thousands of transfers with extremely low associated mortality and is the central component of a high-throughput bioassay system. The gun is constructed...

  14. COLLECTION, CHEMICAL FRACTIONATION, AND MUTAGENICITY BIOASSAY OF AMBIENT AIR PARTICULATE

    EPA Science Inventory

    The influence of industrialization and consequent increased concentration of urban particulate matter on the incidence of cancer has long been a concern. The first bioassays used to evaluate complex ambient air samples were whole-animal carcinogenesis bioassays. In these studies,...

  15. Aspirator gun for high-throughput mosquito bioassays.

    PubMed

    Aldridge, Robert L; Wynn, W Wayne; Britch, Seth C; Linthicum, Kenneth J

    2012-03-01

    We describe an innovative aspirator gun designed to transfer individual anesthetized mosquitoes directly into glass bioassay tubes. The gun has been used for thousands of transfers with extremely low associated mortality and is the central component of a high-throughput bioassay system. The gun is constructed using readily obtainable materials and can be modified for a range of insects. PMID:22533090

  16. Bioassay vs. Air Sampling: Practical Guidance and Experience at Hanford

    SciTech Connect

    Carbaugh, Eugene H.; Carlson, Eric W.; Hill, Robin L.

    2004-02-08

    The Hanford Site has implemented a policy to guide in determining whether air sampling data or special fecal bioassay data are more appropriate for determining doses of record for low-level plutonium exposures. The basis for the policy and four years of experience in comparing DAC-hours exposure with bioassay-based dosimetry is discussed.

  17. Signal Amplification of Bioassay Using Zinc Nanomaterials

    NASA Astrophysics Data System (ADS)

    Cowles, Chad L.

    An emerging trend in the analytical detection sciences is the employment of nanomaterials for bioassay signal transduction to identify analytes critical to public health. These nanomaterials have been specifically investigated for applications which require identification of trace levels of cells, proteins, or other molecules that can have broad ranging impacts to human health in fields such as clinical diagnostics, environmental monitoring, food and drink control, and the prevention of bioterrorism. Oftentimes these nanoparticle-based signal transduction or amplification approaches offer distinct advantages over conventional methods such as increased sensitivity, rapidity, or stability. The biological application of nanoparticles however, does suffer from drawbacks that have limited more widespread adoption of these techniques. Some of these drawbacks are, high cost and toxicity, arduous synthesis methods, functionalization and bioconjugation challenges, and laboratory disposal and environmental hazard issues, all of which have impeded the progression of this technology in some way or another. This work aims at developing novel techniques that offer solutions to a number of these hurdles through the development of new nanoparticle-based signal transduction approaches and the description of a previously undescribed nanomaterial. Zinc-based nanomaterials offer the opportunity to overcome some of the limitations that are encountered when other nanomaterials are employed for bioassay signal transduction. On the other hand, the biological application of zinc nanomaterials has been difficult because in general their fluorescence is in the blue range and the reported quantum yields are usually too low for highly sensitive applications. The advantages of using zinc nanomaterials for biological applications, such as reduced toxicity, simple synthesis, low cost, and straightforward functionalization strategies contribute to the research interest in their application as

  18. Circular Bioassay Platforms for Applications in Microwave-Accelerated Techniques

    PubMed Central

    Mohammed, Muzaffer; Clement, Travis C.; Aslan, Kadir

    2014-01-01

    In this paper, we present the design of four different circular bioassay platforms, which are suitable for homogeneous microwave heating, using theoretical calculations (i.e., COMSOL™ multiphysics software). Circular bioassay platforms are constructed from poly(methyl methacrylate) (PMMA) for optical transparency between 400–800 nm, has multiple sample capacity (12, 16, 19 and 21 wells) and modified with silver nanoparticle films (SNFs) to be used in microwave-accelerated bioassays (MABs). In addition, a small monomode microwave cavity, which can be operated with an external microwave generator (100 W), for use with the bioassay platforms in MABs is also developed. Our design parameters for the circular bioassay platforms and monomode microwave cavity during microwave heating were: (i) temperature profiles, (ii) electric field distributions, (iii) location of the circular bioassay platforms inside the microwave cavity, and (iv) design and number of wells on the circular bioassay platforms. We have also carried out additional simulations to assess the use of circular bioassay platforms in a conventional kitchen microwave oven (e.g., 900 W). Our results show that the location of the circular bioassay platforms in the microwave cavity was predicted to have a significant effect on the homogeneous heating of these platforms. The 21-well circular bioassay platform design in our monomode microwave cavity was predicted to offer a homogeneous heating pattern, where inter-well temperature was observed to be in between 23.72–24.13°C and intra-well temperature difference was less than 0.21°C for 60 seconds of microwave heating, which was also verified experimentally. PMID:25568813

  19. Superluminescent variants of marine luciferases for bioassays.

    PubMed

    Kim, Sung Bae; Suzuki, Hideyuki; Sato, Moritoshi; Tao, Hiroaki

    2011-11-15

    In this study, a rational synthesis of superluminescent variants from marine luciferases with prolonged bioluminescence has been demonstrated. A putative active site of a model marine luciferase, Gaussia princeps Luciferase (GLuc), was assigned and modified by a site-directed mutagenesis. The potent variants were found to generate up to 10 times stronger bioluminescence, emitting red shifts of up to 33 nm with natural coelenterazine than native GLuc, rendering an efficient optical signature in bioassays. The advantageous properties were demonstrated with mammalian two-hybrid assays, single-chain probes, and metastases of murine B16 melanoma in BALB/c nude mice. The unique ideas for engineering GLuc are proved to be valid even for other marine luciferases. PMID:21951281

  20. Modelling larval movement data from individual bioassays.

    PubMed

    McLellan, Chris R; Worton, Bruce J; Deasy, William; Birch, A Nicholas E

    2015-05-01

    We consider modelling the movements of larvae using individual bioassays in which data are collected at a high-frequency rate of five observations per second. The aim is to characterize the behaviour of the larvae when exposed to attractant and repellent compounds. Mixtures of diffusion processes, as well as Hidden Markov models, are proposed as models of larval movement. These models account for directed and localized movements, and successfully distinguish between the behaviour of larvae exposed to attractant and repellent compounds. A simulation study illustrates the advantage of using a Hidden Markov model rather than a simpler mixture model. Practical aspects of model estimation and inference are considered on extensive data collected in a study of novel approaches for the management of cabbage root fly. PMID:25764283

  1. Cell-based bioassays in microfluidic systems

    NASA Astrophysics Data System (ADS)

    Itle, Laura J.; Zguris, Jeanna C.; Pishko, Michael V.

    2004-12-01

    The development of cell-based bioassays for high throughput drug screening or the sensing of biotoxins is contingent on the development of whole cell sensors for specific changes in intracellular conditions and the integration of those systems into sample delivery devices. Here we show the feasibility of using a 5-(and-6)-carboxy SNARF-1, acetoxymethyl ester, acetate, a fluorescent dye capable of responding to changes in intracellular pH, as a detection method for the bacterial endotoxin, lipopolysaccharide. We used photolithography to entrap cells with this dye within poly(ethylene) glyocol diacrylate hydrogels in microfluidic channels. After 18 hours of exposure to lipopolysaccharide, we were able to see visible changes in the fluorescent pattern. This work shows the feasibility of using whole cell based biosensors within microfluidic networks to detect cellular changes in response to exogenous agents.

  2. Genotoxicity of SPL (spent pot lining) as measured by Tradescantia bioassays.

    PubMed

    Andrade-Vieira, L F; Davide, L C; Gedraite, L S; Campos, J M S; Azevedo, H

    2011-10-01

    Spent Pot Liner (SPL) is a solid waste product generated in the process of aluminum production. Tradescantia micronuclei (Trad-MN) and stamen hair mutation (Trad-SHM) bioassays are very useful tests to assess genotoxicity of environmental pollutants. In the present study, we intended to investigate the genotoxicity of this waste with Tradescantia bioassays using leachates of SPL simulating the natural leachability of SPL in soil. The formation of micronuclei (MN) was found to be concentration dependent. MN frequency enhanced significantly with SPL treatment. In addition, SPL also appeared to increase the percentage of dyads and triads. Trad-SHM assay showed that SPL increases pink mutation events as SPL concentration increases. These results demonstrated that SPL is a cytogenotoxic agent that affects different genetic end-points (induction of micronuclei and point mutations) even at low concentration (2% and 3%). PMID:21802142

  3. Plasmonically amplified bioassay - Total internal reflection fluorescence vs. epifluorescence geometry.

    PubMed

    Hageneder, Simone; Bauch, Martin; Dostalek, Jakub

    2016-08-15

    This paper investigates plasmonic amplification in two commonly used optical configurations for fluorescence readout of bioassays - epifluorescence (EPF) and total internal reflection fluorescence (TIRF). The plasmonic amplification in the EPF configuration was implemented by using crossed gold diffraction grating and Kretschmann geometry of attenuated total reflection method (ATR) was employed in the TIRF configuration. Identical assay, surface architecture for analyte capture, and optics for the excitation, collection and detection of emitted fluorescence light intensity were used in both TIRF and EPF configurations. Simulations predict that the crossed gold diffraction grating (EPF) can amplify the fluorescence signal by a factor of 10(2) by the combination of surface plasmon-enhanced excitation and directional surface plasmon-coupled emission in the red part of spectrum. This factor is about order of magnitude higher than that predicted for the Kretschmann geometry (TIRF) which only took advantage of the surface plasmon-enhanced excitation. When applied for the readout of sandwich interleukin 6 (IL-6) immunoassay, the plasmonically amplified EPF geometry designed for Alexa Fluor 647 labels offered 4-times higher fluorescence signal intensity compared to TIRF. Interestingly, both geometries allowed reaching the same detection limit of 0.4pM despite of the difference in the fluorescence signal enhancement. This is attributed to inherently lower background of fluorescence signal for TIRF geometry compared to that for EPF which compensates for the weaker fluorescence signal enhancement. The analysis of the inflammation biomarker IL-6 in serum at medically relevant concentrations and the utilization of plasmonic amplification for the fluorescence measurement of kinetics of surface affinity reactions are demonstrated for both EPF and TIRF readout. PMID:27260457

  4. PubChem BioAssay: 2014 update.

    PubMed

    Wang, Yanli; Suzek, Tugba; Zhang, Jian; Wang, Jiyao; He, Siqian; Cheng, Tiejun; Shoemaker, Benjamin A; Gindulyte, Asta; Bryant, Stephen H

    2014-01-01

    PubChem's BioAssay database (http://pubchem.ncbi.nlm.nih.gov) is a public repository for archiving biological tests of small molecules generated through high-throughput screening experiments, medicinal chemistry studies, chemical biology research and drug discovery programs. In addition, the BioAssay database contains data from high-throughput RNA interference screening aimed at identifying critical genes responsible for a biological process or disease condition. The mission of PubChem is to serve the community by providing free and easy access to all deposited data. To this end, PubChem BioAssay is integrated into the National Center for Biotechnology Information retrieval system, making them searchable by Entrez queries and cross-linked to other biomedical information archived at National Center for Biotechnology Information. Moreover, PubChem BioAssay provides web-based and programmatic tools allowing users to search, access and analyze bioassay test results and metadata. In this work, we provide an update for the PubChem BioAssay resource, such as information content growth, new developments supporting data integration and search, and the recently deployed PubChem Upload to streamline chemical structure and bioassay submissions. PMID:24198245

  5. PubChem BioAssay: 2014 update

    PubMed Central

    Wang, Yanli; Suzek, Tugba; Zhang, Jian; Wang, Jiyao; He, Siqian; Cheng, Tiejun; Shoemaker, Benjamin A.; Gindulyte, Asta; Bryant, Stephen H.

    2014-01-01

    PubChem’s BioAssay database (http://pubchem.ncbi.nlm.nih.gov) is a public repository for archiving biological tests of small molecules generated through high-throughput screening experiments, medicinal chemistry studies, chemical biology research and drug discovery programs. In addition, the BioAssay database contains data from high-throughput RNA interference screening aimed at identifying critical genes responsible for a biological process or disease condition. The mission of PubChem is to serve the community by providing free and easy access to all deposited data. To this end, PubChem BioAssay is integrated into the National Center for Biotechnology Information retrieval system, making them searchable by Entrez queries and cross-linked to other biomedical information archived at National Center for Biotechnology Information. Moreover, PubChem BioAssay provides web-based and programmatic tools allowing users to search, access and analyze bioassay test results and metadata. In this work, we provide an update for the PubChem BioAssay resource, such as information content growth, new developments supporting data integration and search, and the recently deployed PubChem Upload to streamline chemical structure and bioassay submissions. PMID:24198245

  6. Collection and control of tritium bioassay samples at Pantex

    SciTech Connect

    Fairrow, N.L.; Ivie, W.E.

    1992-01-01

    Pantex is the final assembly/disassembly point for US nuclear weapons. The Pantex internal dosimetry section monitors radiation workers once a month for tritium exposure. In order to manage collection and control of the bioassay specimens efficiently, a bar code system for collection of samples was developed and implemented to speed up the process and decrease the number of errors probable when transferring data. In the past, all the bioassay data from samples were entered manually into a computer database. Transferring the bioassay data from the liquid scintillation counter to each individual's dosimetry record required as much as two weeks of concentrated effort.

  7. Collection and control of tritium bioassay samples at Pantex

    SciTech Connect

    Fairrow, N.L.; Ivie, W.E.

    1992-12-31

    Pantex is the final assembly/disassembly point for US nuclear weapons. The Pantex internal dosimetry section monitors radiation workers once a month for tritium exposure. In order to manage collection and control of the bioassay specimens efficiently, a bar code system for collection of samples was developed and implemented to speed up the process and decrease the number of errors probable when transferring data. In the past, all the bioassay data from samples were entered manually into a computer database. Transferring the bioassay data from the liquid scintillation counter to each individual`s dosimetry record required as much as two weeks of concentrated effort.

  8. Estrogen Receptor Agonists and Antagonists in the Yeast Estrogen Bioassay.

    PubMed

    Wang, Si; Bovee, Toine F H

    2016-01-01

    Cell-based bioassays can be used to predict the eventual biological activity of a substance on a living organism. In vitro reporter gene bioassays are based on recombinant vertebrate cell lines or yeast strains and especially the latter are easy-to-handle, cheap, and fast. Moreover, yeast cells do not express estrogen, androgen, progesterone or glucocorticoid receptors, and are thus powerful tools in the development of specific reporter gene systems that are devoid of crosstalk from other hormone pathways. This chapter describes our experience with an in-house developed RIKILT yeast estrogen bioassay for testing estrogen receptor agonists and antagonists, focusing on the applicability of the latter. PMID:26585147

  9. Evaluation of a feline-specific multiplex, bead-based assay for detection of cytokines, chemokines, growth factors, and other immunologically active proteins in serum and plasma samples from cats.

    PubMed

    Halpin, Rachel E; Saunders, Rebecca S; Thompson, Beverly J; Rohde Newgent, Allison S; Amorim, Juliana; Melillo, Gabrielle N; DeClue, Amy E

    2016-05-01

    OBJECTIVE To evaluate a feline-specific multiplex, bead-based assay system for detection of recombinant and native proteins in serum samples and in EDTA-treated and heparinized plasma samples. SAMPLE Serum samples and EDTA-treated and heparinized plasma samples from 30 sick cats and 9 healthy client-owned cats and heparinized whole blood samples from 5 healthy purpose-bred cats. PROCEDURES Ability of the assay system to detect 19 recombinant and native immunologically active proteins in plasma and serum samples from healthy and purpose-bred cats was evaluated via spike-and-recovery tests, assessments of inter- and intra-assay variation, linearity results, and leukocyte stimulation. Effects of various concentrations of heparin and serum matrix solution on percentages of analytes recovered were also evaluated. Analyte concentrations in samples from healthy and sick cats were measured and compared between groups. RESULTS Percentages of analytes recovered were unsatisfactory for most assays. Serum and heparinized plasma samples yielded better recovery results than did EDTA-treated plasma samples. Use of serum matrix solution did not improve results. Use of heparin concentrations greater than the recommended range affected the results. Linearity of results was difficult to assess because of the poor recovery. For the analytes that were recovered sufficiently for assessment, linearity appeared to be reasonable despite the limited detection. CONCLUSIONS AND CLINICAL RELEVANCE Poor percentages of analytes recovered and adverse effects of sample protein matrix limited the usefulness of the multiplex, bead-based assay system for measurement of immunologically active proteins in solutions with high protein content; however, recovery results were fairly linear, potentially allowing evaluation of feline plasma or serum samples with high analyte concentrations. PMID:27111017

  10. Long-wavelength-emitting nanocrystals for bioassay applications

    NASA Astrophysics Data System (ADS)

    Leppert, Valerie J.; Harvey, Ashley S.; McCool, Geoff D.; Quinlan, Forest T.; Feng, Jun; Shan, Guomin; Stroeve, Pieter; Risbud, Subhash H.; Hammock, Bruce D.; Kennedy, Ian M.

    2002-11-01

    New fluorophores that can be excited using visible or near-infrared radiation are of considerable interest for application in environmental and complex bioassays, where background fluorescence is exacerbated by ultra-violet or blue excitation. Useful labels for biomolecules include infrared emitting semiconductor nanoparticles that can be blue-shifted into the near-infrared and visible through quantum confinement effects, oxides of iron, and rare earth oxides. In this work, the synthesis of 6 nm average diameter lead selenide nanocrystals (well below the Bohr exciton diameter of 92 nm) through a reverse micelle technique; and the synthesis of iron and europium oxides with particles less than 5 nm in diameter by pulsed laser ablation is reported. The europium oxide nanoparticles' emission showed a large Stokes shift (144 nm or 216 nm, depending on excitation wavelength); a narrow, symmetric emission line at 610 nm (FWHM of 8 nm); and long lifetime (300 μs). The Eu2O3 nanoparticles, which were coated with silica for functionalization, displayed a greatly enhanced sensitivity over a conventional ELISA (0.025 ng ml-1 vs. 0.1 ng ml-1) when run in an atrazine immunoassay.

  11. Luminescent Lanthanide Reporters for High-Sensitivity Novel Bioassays.

    SciTech Connect

    Anstey, Mitchell; Fruetel, Julia A.; Foster, Michael E.; Hayden, Carl C.; Buckley, Heather L.; Arnold, John

    2013-09-01

    Biological imaging and assay technologies rely on fluorescent organic dyes as reporters for a number of interesting targets and processes. However, limitations of organic dyes such as small Stokes shifts, spectral overlap of emission signals with native biological fluorescence background, and photobleaching have all inhibited the development of highly sensitive assays. To overcome the limitations of organic dyes for bioassays, we propose to develop lanthanide-based luminescent dyes and demonstrate them for molecular reporting applications. This relatively new family of dyes was selected for their attractive spectral and chemical properties. Luminescence is imparted by the lanthanide atom and allows for relatively simple chemical structures that can be tailored to the application. The photophysical properties offer unique features such as narrow and non-overlapping emission bands, long luminescent lifetimes, and long wavelength emission, which enable significant sensitivity improvements over organic dyes through spectral and temporal gating of the luminescent signal.Growth in this field has been hindered due to the necessary advanced synthetic chemistry techniques and access to experts in biological assay development. Our strategy for the development of a new lanthanide-based fluorescent reporter system is based on chelation of the lanthanide metal center using absorbing chromophores. Our first strategy involves %22Click%22 chemistry to develop 3-fold symmetric chelators and the other involves use of a new class of tetrapyrrole ligands called corroles. This two-pronged approach is geared towards the optimization of chromophores to enhance light output.

  12. Evaporation-Driven Bioassays in Suspended Droplets.

    PubMed

    Hernandez-Perez, Ruth; Fan, Z Hugh; Garcia-Cordero, Jose L

    2016-07-19

    The microtiter plate has been an essential tool for diagnostics, high-throughput screening, and biological assays. We present an alternative platform to perform bioassays in a microplate format that exploits evaporation to drive assay reactions. Our method consists of droplets suspended on plastic pillars; reactions occur in these droplets instead of the wells. The pillars are fabricated by milling, and the rough surface created by this fabrication method pins the droplet to a constant contact line during the assay and also acts as a hydrophobic surface. Upon evaporation, natural convection arising from Marangoni currents mixes solutions in the droplet, which speeds up assay reactions, decreases assay times, and increases limits of detection. As a proof of concept we implemented two colorimetric assays to detect glucose and proteins in only 1.5 μL, without any external devices for mixing and with a digital microscope as a readout mechanism. Our platform is an ideal alternative to the microtiter plate, works with different volumes, is compatible with commercially available reagent dispensers and plate-readers, and could have broad applications in diagnostics and high-throughput screening. PMID:27331825

  13. Annotating Human P-Glycoprotein Bioassay Data

    PubMed Central

    Zdrazil, Barbara; Pinto, Marta; Vasanthanathan, Poongavanam; Williams, Antony J; Balderud, Linda Zander; Engkvist, Ola; Chichester, Christine; Hersey, Anne; Overington, John P; Ecker, Gerhard F

    2012-01-01

    Abstract Huge amounts of small compound bioactivity data have been entering the public domain as a consequence of open innovation initiatives. It is now the time to carefully analyse existing bioassay data and give it a systematic structure. Our study aims to annotate prominent in vitro assays used for the determination of bioactivities of human P-glycoprotein inhibitors and substrates as they are represented in the ChEMBL and TP-search open source databases. Furthermore, the ability of data, determined in different assays, to be combined with each other is explored. As a result of this study, it is suggested that for inhibitors of human P-glycoprotein it is possible to combine data coming from the same assay type, if the cell lines used are also identical and the fluorescent or radiolabeled substrate have overlapping binding sites. In addition, it demonstrates that there is a need for larger chemical diverse datasets that have been measured in a panel of different assays. This would certainly alleviate the search for other inter-correlations between bioactivity data yielded by different assay setups. PMID:23293680

  14. Bioassay-Directed Fractionation of Diesel and Biodiesel Emissions

    EPA Science Inventory

    Biofuels are being developed as alternatives to petroleum-derived products, but published research is contradictory regarding the mutagenic activity of such emissions relative to those from petroleum diesel. We performed bioassay-directed fractionation and analyzed the polycyclic...

  15. Bioassay Phantoms Using Medical Images and Computer Aided Manufacturing

    SciTech Connect

    Dr. X. Geroge Xu

    2011-01-28

    A radiation bioassay program relies on a set of standard human phantoms to calibrate and assess radioactivity levels inside a human body for radiation protection and nuclear medicine imaging purposes. However, the methodologies in the development and application of anthropomorphic phantoms, both physical and computational, had mostly remained the same for the past 40 years. We herein propose a 3-year research project to develop medical image-based physical and computational phantoms specifically for radiation bioassay applications involving internally deposited radionuclides. The broad, long-term objective of this research was to set the foundation for a systematic paradigm shift away from the anatomically crude phantoms in existence today to realistic and ultimately individual-specific bioassay methodologies. This long-term objective is expected to impact all areas of radiation bioassay involving nuclear power plants, U.S. DOE laboratories, and nuclear medicine clinics.

  16. A CONTROLLED BIOASSAY SYSTEM FOR MEASURING TOXICITY OF HEAVY METALS

    EPA Science Inventory

    Biological availability of metal micronutrients and metal toxicity are believed to be dependent on metal oxidation state, complexation, and solubility as well as the physicochemical characteristics of the aqueous phase. Basic design criteria for fish bioassays which are capable o...

  17. A wind tunnel bioassay system for screening mosquito repellents.

    PubMed

    Sharpington, P J; Healy, T P; Copland, M J

    2000-09-01

    A wind tunnel bioassay system to screen mosquito repellents is described. A wind tunnel is utilized to exploit the upwind flight response of host-seeking mosquitoes. Mosquitoes within the wind tunnel are activated with human breath, fly upwind, and land on heated chick skins. This behavioral sequence results in a consistently high percentage of the test population approaching repellent or control stimuli. The bioassay system is calibrated with diethyl methylbenzamide against Aedes aegypti and demonstrates a reproducible dose-response relationship. The persistence of diethyl methyl benzamide after a 1-h period is also recorded. The design of the bioassay system permits simultaneous, independent testing of 3 candidate repellents. The wind tunnel bioassay system is compared to other techniques for evaluating mosquito repellents. PMID:11081652

  18. In Vitro Androgen Bioassays as a Detection Method for Designer Androgens

    PubMed Central

    Cooper, Elliot R.; McGrath, Kristine C. Y.; Heather, Alison K.

    2013-01-01

    Androgens are the class of sex steroids responsible for male sexual characteristics, including increased muscle mass and decreased fat mass. Illicit use of androgen doping can be an attractive option for those looking to enhance sporting performance and/or physical appearance. The use of in vitro bioassays to detect androgens, especially designer or proandrogens, is becoming increasingly important in combating androgen doping associated with nutritional supplements. The nutritional sports supplement market has grown rapidly throughout the past decade. Many of these supplements contain androgens, designer androgens or proandrogens. Many designer or proandrogens cannot be detected by the standard highly-sensitive screening methods such as gas chromatography-mass spectrometry because their chemical structure is unknown. However, in vitro androgen bioassays can detect designer and proandrogens as these assays are not reliant on knowing the chemical structure but instead are based on androgen receptor activation. For these reasons, it may be advantageous to use routine androgen bioassay screening of nutraceutical samples to help curb the increasing problem of androgen doping. PMID:23389345

  19. A novel bronchial ring bioassay for the evaluation of small airway smooth muscle function in mice.

    PubMed

    Liu, John Q; Yang, Dennis; Folz, Rodney J

    2006-08-01

    Advances in our understanding of murine airway physiology have been hindered by the lack of suitable, ex vivo, small airway bioassay systems. In this study, we introduce a novel small murine airway bioassay system that permits the physiological and pharmacological study of intrapulmonary bronchial smooth muscle via a bronchial ring (BR) preparation utilizing BR segments as small as 200 microm in diameter. Using this ex vivo BR bioassay, we characterized small airway smooth muscle contraction and relaxation in the presence and absence of bronchial epithelium. In control BRs, the application of mechanical stretch is followed by spontaneous bronchial smooth muscle relaxation. BRs pretreated with methacholine (MCh) partially attenuate this stretch-induced relaxation by as much as 42% compared with control. MCh elicited a dose-dependent bronchial constriction with a maximal tension (E(max)) of 8.7 +/- 0.2 mN at an EC(50) of 0.33 +/- 0.02 microM. In the presence of nifedipine, ryanodine, 2-aminoethoxydiphenyl borate, and SKF-96365, E(max) to MCh was significantly reduced. In epithelium-denuded BRs, MCh-induced contraction was significantly enhanced to 11.4 +/- 1.0 mN with an EC(50) of 0.16 +/- 0.04 microM (P < 0.01). Substance P relaxed MCh-precontracted BR by 62.1%; however, this bronchial relaxation effect was completely lost in epithelium-denuded BRs. Papaverine virtually abolished MCh-induced constriction in both epithelium-intact and epithelium-denuded bronchial smooth muscle. In conclusion, this study introduces a novel murine small airway BR bioassay that allows for the physiological study of smooth muscle airway contractile responses that may aid in our understanding of the pathophysiology of asthma. PMID:16648239

  20. Comparison of laboratory batch and flow-through microcosm bioassays.

    PubMed

    Clément, Bernard J P; Delhaye, Hélène L; Triffault-Bouchet, Gaëlle G

    2014-10-01

    Since 1997, we have been developing a protocol for ecotoxicological bioassays in 2-L laboratory microcosms and have applied it to the study of various pollutants and ecotoxicological risk assessment scenarios in the area of urban facilities and transport infrastructures. The effects on five different organisms (micro-algae, duckweeds, daphnids, amphipods, chironomids) are assessed using biological responses such as growth, emergence (chironomids), reproduction (daphnids) and survival, with a duration of exposure of 3 weeks. This bioassay has mainly been used as a batch bioassay, i.e., the water was not renewed during the test. A flow-through microcosm bioassay has been developed recently, with the assumption that conditions for the biota should be improved, variability reduced, and the range of exposure patterns enlarged (e.g., the possibility of maintaining constant exposure in the water column). This paper compares the results obtained in batch and flow-through microcosm bioassays, using cadmium as a model toxicant. As expected, the stabilization of physico-chemical parameters, increased organism fitness and reduced variability were observed in the flow-through microcosm bioassay. PMID:25086825

  1. Primary Bioassay of Human Myeloma Stem Cells

    PubMed Central

    Hamburger, Anne; Salmon, Sydney E.

    1977-01-01

    The ability to clone primary tumors in soft agar has proven useful in the study of the kinetics and biological properties of tumor stem cells. We report the development of an in vitro assay which permits formation of colonies of human monoclonal plasma cells in soft agar. Colony growth has been observed from bone marrow aspirates from 75% of the 70 patients with multiple myeloma or related monoclonal disorders studied. Growth was induced with either 0.02 ml of human type O erythrocytes or 0.25 ml of medium conditioned by the adherent spleen cells of mineral oil-primed BALB/c mice. 5-500 colonies appeared after 2-3 wk in culture yielding a plating efficiency of 0.001-0.1%. The number of myeloma colonies was proportional to the number of cells plated between concentrations of 105-106 and back-extrapolated through zero, suggesting that colonies were clones derived from single myeloma stem cells. Morphological, histochemical, and functional criteria showed the colonies to consist of immature plasmablasts and mature plasma cells. 60-80% of cells picked from colonies contained intracytoplasmic monoclonal immunoglobulin. Colony growth was most easily achieved from the bone marrow cells of untreated patients or those in relapse. Only 50% of bone marrow samples from patients in remission were successfully cultured. Tritiated thymidine suicide studies provided evidence that for most myeloma patients, a very high proportion of myeloma colony-forming cells was actively in transit through the cell cycle. Velocity sedimentation at 1 g showed myeloma stem cells sedimented in a broad band with a peak at 13 mm/h. Antibody to granulocyte colony-stimulating factor did not reduce the number or size of the colonies. Increased numbers of myeloma colonies were seen when the marrow was depleted of colony-stimulating factor elaborating adherent cells before plating. This bioassay should prove useful in studying the in vitro biological behavior of certain bone marrow-derived (B

  2. A novel laboratory screening bioassay for crop seedling allelopathy.

    PubMed

    Belz, Regina G; Hurle, Karl

    2004-01-01

    Crops that control weeds by root exudation of allelochemicals are receiving increased attention, and there are efforts to breed allelopathic cultivars in several crops. The genetic improvement of allelopathic traits is based upon parental germ plasm with high allelopathic activity. Identification of allelopathic germplasm is done in laboratory screening bioassays, but experimental protocols are limited. We developed a fast and reliable laboratory screening bioassay for grain crops that includes dose-response considerations as an integral part of the experimental design. The bioassay was conducted in hydroponic culture, and a range of experiments with 2-(3H)-benzoxazolinone (BOA), an allelochemical of several grain crops, was carried out to define the basic protocol. Because of its sensitivity to BOA, Sinapis alba L. was selected as the receiver species. BOA affected growth (fresh weight and length of shoot and root), enzyme activities (ascorbate peroxidase, catalase, glutathione S-transferase, peroxidase, phenylalanine ammonia-lyase), and chlorophyll fluorescence, whereby root length was the most reliable response parameter. BOA sensitivity was dependent on nutrients for all parameters measured, and, thus, no nutrients were added. A set of experiments with Secale cereale L. and Triticum aestivum L. as donor species was carried out to optimize the protocol. Light and pH were eliminated as primary causes for the observed inhibition. The proposed bioassay has several methodological advantages over current bioassays. PMID:15074665

  3. Soil bioassays and the {sup 129}I problem

    SciTech Connect

    Sheppard, S.C.

    1995-12-31

    Iodine-129 is a very long-lived radionuclide associated with spent nuclear fuel. Because {sup 129}I has a 10{sup 7}-year half-life, is very mobile in the environment and is a biologically essential element, it is the most limiting radionuclide affecting disposal of spent fuel. Traditionally, the potential impacts of {sup 129}I have been estimated for human receptors, with the implicit assumption that all other organisms are less at risk. Risk is the operative word, the objective for protection of humans is to protect individuals, whereas the objective for other biota is usually to protect populations. Here, {sup 129}I poses an interesting problem: the half-life is so long it is barely radioactive. Thus, the chemical toxicity may be more limiting than the radiological impact. A series of soil bioassays were employed, including a life-cycle plant (Brassica rapa) bioassay, a modified earthworm survival bioassay, a microarthropod colonization/survival bioassay, and a series of more common soil and aquatic bioassays. Chemical toxicity was indicated at soil concentrations as low as 5 mg kg{sup {minus}1}. At these levels, radiological impact on non-human biota would not be expected, and therefore the chemical toxicity effects are more critical. However, human food-chain model estimates show these levels, as pure {sup 129}I, would be unacceptable for human radiological exposure, so that for {sup 129}I, protection of the human environment should also be protective of non-human biota.

  4. [Investigation on pattern and methods of quality control for Chinese materia medica based on dao-di herbs and bioassay - bioassay for Coptis chinensis].

    PubMed

    Yan, Dan; Xiao, Xiao-he

    2011-05-01

    Establishment of bioassay methods is the technical issues to be faced with in the bioassay of Chinese materia medica. Taking the bioassay of Coptis chinensis Franch. as an example, the establishment process and application of the bioassay methods (including bio-potency and bio-activity fingerprint) were explained from the aspects of methodology, principle of selection, experimental design, method confirmation and data analysis. The common technologies were extracted and formed with the above aspects, so as to provide technical support for constructing pattern and method of the quality control for Chinese materia medica based on the dao-di herbs and bioassay. PMID:21800546

  5. Environmental effects of dredging. A chronic sublethal sediment bioassay with the marine polychaete nereis (Neanthes) arenaceodentata

    SciTech Connect

    Dillon, T.M.; Moore, D.W.; Bridges, T.S.

    1995-01-01

    This note provides a general overview of a new 28-day chronic sublethal sediment bioassay designed for the regulatory evaluation of dredged material. The bioassay uses survival and growth rate endpoints with the polychaete Nereis (Neanthes) arenaceodentata. The primary technical reference for this new bioassay is Dillon, Moore, and Reish (in press), upon which this overview is based. Sediment bioassays are used to assess the aggregate toxicity of sediment associated anthropogenic chemicals. Historically, these bioassays have measured survival of highly sensitive species following acute exposures (10 days). A new generation of sediment bioassays is being developed in which the subtle, sublethal response of test species is measured following chronic sediment exposures (Dillon 1993).

  6. Carbon-14 Bioassay for Decommissioning of Hanford Reactors

    SciTech Connect

    Carbaugh, Eugene H.; Watson, David J.

    2012-05-01

    The old production reactors at the US Department of Energy Hanford Site used large graphite piles as the moderator. As part of long-term decommissioning plans, the potential need for 14C radiobioassay of workers was identified. Technical issues associated with 14C bioassay and worker monitoring were investigated, including anticipated graphite characterization, potential intake scenarios, and the bioassay capabilities that may be required to support the decommissioning of the graphite piles. A combination of urine and feces sampling would likely be required for the absorption type S 14C anticipated to be encountered. However the concentrations in the graphite piles appear to be sufficiently low that dosimetrically significant intakes of 14C are not credible, thus rendering moot the need for such bioassay.

  7. Method comparison for 241Am emergency urine bioassay.

    PubMed

    Li, Chunsheng; Sadi, Baki; Benkhedda, Karima; St-Amant, Nadereh; Moodie, Gerry; Ko, Raymond; Dinardo, Anthony; Kramer, Gary

    2010-10-01

    241Am is one of the high-risk radionuclides that might be used in a terrorist attack. 241Am in urine bioassay can identify the contaminated individuals who need immediate medical intervention and decontamination. This paper compares three methods for the measurement of 241Am in urine, namely liquid scintillation counting (LSC), inductively coupled plasma mass spectrometry (ICP-MS) and gamma spectrometry (GS), at two levels, 20 and 2 Bq l(-1). All three methods satisfied the ANSI N13.30 radio-bioassay criteria for accuracy and repeatability. ICP-MS offered the best sensitivity and fastest sample turnaround; however, the ICP-MS system used in this work may not be available in many bioassay laboratories. LSC and GS are more commonly available instruments. GS requires minimal or no sample preparation, which makes it a good candidate method. Moreover, the sample throughput can be significantly improved if the GS and LSC methods are automated. PMID:20573683

  8. Internal dosimetry performing dose assessments via bioassay measurements

    SciTech Connect

    Bailey, K.M.

    1993-05-11

    The Internal Dosimetry Department at the Y-12 Plant maintains a state-of-the-art bioassay program managed under the guidance and regulations of the Department of Energy. The two major bioassay techniques currently used at Y-12 are the in vitro (urinalysis) and in vivo (lung counting) programs. Fecal analysis (as part of the in vitro program) is another alternative; however, since both urine and fecal analysis provide essentially the same capabilities for detecting exposures to uranium, the urinalysis is the main choice primarily for aesthetic reasons. The bioassay frequency is based on meeting NCRP 87 objectives which are to monitor the accumulation of radioactive material in exposed individuals, and to ensure that significant depositions are detected.

  9. The effect of pesticide residue on caged mosquito bioassays.

    PubMed

    Barber, J A S; Greer, Mike; Coughlin, Jamie

    2006-09-01

    Wind tunnel experiments showed that secondary pickup of insecticide residue by mosquitoes in cage bioassays had a significant effect on mortality. Cage bioassays using adult Ochlerotatus taeniorhynchus (Wiedemann) investigated the effect of exposure time to a contaminated surface. Cages were dosed in a wind tunnel using the LC50 for naled (0.124 mg a.i./ml) and an LC25 (0.0772 mg a.i./ml) for naled. Half of the bioassay mosquitoes were moved directly into clean cages with the other half remaining in the sprayed, hence contaminated, cage. Treatment mortality was assessed at 8, 15, 30, 60, 120, 240, and 1,440 min postapplication. Cage contamination had a significant effect on mosquito mortality for both the LC25 and LC50 between 15 and 30 min postapplication. PMID:17067048

  10. Bioassay-directed chemical analysis in environmental research

    SciTech Connect

    Schuetzle, D.; Lewtas, J.

    1986-01-01

    The use of short-term bioassay tests in conjunction with analytical measurements, constitute a powerful tool for identifying important environmental contaminants. The authors have coined the terminology bioassay directed chemical analysis to best describe this marriage of analytical chemistry and biology. The objective of this methodology is to identify key compounds in various types of air-pollutant samples. Once that task is completed, studies on metabolism, sources, environmental exposure and atmospheric chemistry can be undertaken. The principles and methodologies for bioassay directed chemical analysis are presented and illustrated in this paper. Most of this work has been directed toward the characterization of ambient air and diesel particulates, which are used as examples in this report to illustrate the analytical logic used for identifying the bio-active components of complex mixtures.

  11. Do we really need in-situ bioassays?

    SciTech Connect

    Salazar, M.H.; Salazar, S.M.

    1995-12-31

    In-situ bioassays are needed to validate the results from laboratory testing and to understand biological interactions. Standard laboratory protocols provide reproducible test results, and the precision of those tests can be mathematically defined. Significant correlations between toxic substances and levels of response (bioaccumulation and bioeffects) have also been demonstrated with natural field populations and suggest that the laboratory results can accurately predict field responses. An equal number of studies have shown a lack of correlation between laboratory bioassay results and responses of natural field populations. The best way to validate laboratory results is with manipulative field testing; i.e., in-situ bioassays with caged organisms. Bioaccumulation in transplanted bivalves has probably been the most frequently used form of an in-situ bioassay. The authors have refined those methods to include synoptic measurements of bioaccumulation and growth. Growth provides an easily-measured bioeffects endpoint and a means of calibrating bioaccumulation. Emphasis has been on minimizing the size range of test animals, repetitive measurements of individuals and standardization of test protocols for a variety of applications. They are now attempting to standardize criteria for accepting and interpreting data in the same way that laboratory bioassays have been standardized. Others have developed methods for in-situ bioassays using eggs, larvae, unicellular organisms, crustaceans, benthic invertebrates, bivalves, and fish. In the final analysis, the in-situ approach could be considered as an exposure system where any clinical measurements are possible. The most powerful approach would be to use the same species in laboratory and field experiments with the same endpoints.

  12. A statistical treatment of bioassay pour fractions

    NASA Astrophysics Data System (ADS)

    Barengoltz, Jack; Hughes, David

    A bioassay is a method for estimating the number of bacterial spores on a spacecraft surface for the purpose of demonstrating compliance with planetary protection (PP) requirements (Ref. 1). The details of the process may be seen in the appropriate PP document (e.g., for NASA, Ref. 2). In general, the surface is mechanically sampled with a damp sterile swab or wipe. The completion of the process is colony formation in a growth medium in a plate (Petri dish); the colonies are counted. Consider a set of samples from randomly selected, known areas of one spacecraft surface, for simplicity. One may calculate the mean and standard deviation of the bioburden density, which is the ratio of counts to area sampled. The standard deviation represents an estimate of the variation from place to place of the true bioburden density commingled with the precision of the individual sample counts. The accuracy of individual sample results depends on the equipment used, the collection method, and the culturing method. One aspect that greatly influences the result is the pour fraction, which is the quantity of fluid added to the plates divided by the total fluid used in extracting spores from the sampling equipment. In an analysis of a single sample’s counts due to the pour fraction, one seeks to answer the question: What is the probability that if a certain number of spores are counted with a known pour fraction, that there are an additional number of spores in the part of the rinse not poured. This is given for specific values by the binomial distribution density, where detection (of culturable spores) is success and the probability of success is the pour fraction. A special summation over the binomial distribution, equivalent to adding for all possible values of the true total number of spores, is performed. This distribution when normalized will almost yield the desired quantity. It is the probability that the additional number of spores does not exceed a certain value. Of course

  13. A statistical treatment of bioassay pour fractions

    NASA Astrophysics Data System (ADS)

    Barengoltz, Jack; Hughes, David

    A bioassay is a method for estimating the number of bacterial spores on a spacecraft surface for the purpose of demonstrating compliance with planetary protection (PP) requirements (Ref. 1). The details of the process may be seen in the appropriate PP document (e.g., for NASA, Ref. 2). In general, the surface is mechanically sampled with a damp sterile swab or wipe. The completion of the process is colony formation in a growth medium in a plate (Petri dish); the colonies are counted. Consider a set of samples from randomly selected, known areas of one spacecraft surface, for simplicity. One may calculate the mean and standard deviation of the bioburden density, which is the ratio of counts to area sampled. The standard deviation represents an estimate of the variation from place to place of the true bioburden density commingled with the precision of the individual sample counts. The accuracy of individual sample results depends on the equipment used, the collection method, and the culturing method. One aspect that greatly influences the result is the pour fraction, which is the quantity of fluid added to the plates divided by the total fluid used in extracting spores from the sampling equipment. In an analysis of a single sample’s counts due to the pour fraction, one seeks to answer the question: What is the probability that if a certain number of spores are counted with a known pour fraction, that there are an additional number of spores in the part of the rinse not poured. This is given for specific values by the binomial distribution density, where detection (of culturable spores) is success and the probability of success is the pour fraction. A special summation over the binomial distribution, equivalent to adding for all possible values of the true total number of spores, is performed. This distribution when normalized will almost yield the desired quantity. It is the probability that the additional number of spores does not exceed a certain value. Of course

  14. HIGHLY SENSITIVE BIOASSAYS FOR EVALUATING AIRBORNE MUTAGENS INDOORS

    EPA Science Inventory

    The standard mutagenicity bioassays that are readily applied to the valuation of outdoor air samples collected by high volume samplers are not efficiently sensitive to measure the mutagenicity of low volume air samples collected indoors. wo microsuspension mutation assays using v...

  15. Filtration effects due to bioassay cage design and screen type

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The use of bioassay cages in the efficacy assessment of specific compounds, application techniques and technologies is a common practice. There are a number of cage designs being used that range across a variety of cage shapes and sizes and mesh types. The objective of this work was to examine a r...

  16. STRESS ETHYLENE: A BIOASSAY FOR RHIZOSPHERE-APPLIED PHYTOTOXICANTS

    EPA Science Inventory

    A bioassay for rhizosphere-applied phytotoxicants was developed and evaluated with a broad range of chemicals. Test substances were applied to the rhizosphere of whole, intact bush bean plants (Phaseolus vulgaris L. cv. Bush Blue Lake 290) grown in a solid support medium and the ...

  17. Assessment of acrylamide toxicity using a battery of standardised bioassays.

    PubMed

    Zovko, Mira; Vidaković-Cifrek, Željka; Cvetković, Želimira; Bošnir, Jasna; Šikić, Sandra

    2015-12-01

    Acrylamide is a monomer widely used as an intermediate in the production of organic chemicals, e.g. polyacrylamides (PAMs). Since PAMs are low cost chemicals with applications in various industries and waste- and drinking water treatment, a certain amount of non-polymerised acrylamide is expected to end up in waterways. PAMs are non-toxic but acrylamide induces neurotoxic effects in humans and genotoxic, reproductive, and carcinogenic effects in laboratory animals. In order to evaluate the effect of acrylamide on freshwater organisms, bioassays were conducted on four species: algae Desmodesmus subspicatus and Pseudokirchneriella subcapitata, duckweed Lemna minor and water flea Daphnia magna according to ISO (International Organization for Standardisation) standardised methods. This approach ensures the evaluation of acrylamide toxicity on organisms with different levels of organisation and the comparability of results, and it examines the value of using a battery of low-cost standardised bioassays in the monitoring of pollution and contamination of aquatic ecosystems. These results showed that EC50 values were lower for Desmodesmus subspicatus and Pseudokirchneriella subcapitata than for Daphnia magna and Lemna minor, which suggests an increased sensitivity of algae to acrylamide. According to the toxic unit approach, the values estimated by the Lemna minor and Daphnia magna bioassays, classify acrylamide as slightly toxic (TU=0-1; Class 1). The results obtained from algal bioassays (Desmodesmus subspicatus and Pseudokirchneriella subcapitata) revealed the toxic effect of acrylamide (TU=1-10; Class 2) on these organisms. PMID:26751864

  18. Artificial diets for life tables bioassays of TPB in Mississippi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two artificial diets for mass rearing and bioassay of the tarnished plant bug, (TPB), Lygus lineolaris Palisot de Beauvois, (Hemiptera: Miridae) were modified and developed, respectively. The first diet is a modification of a semisolid artificial diet (NI diet), which permits large scale rearing of ...

  19. INFLUENCE OF SEDIMENT EXTRACT FRACTIONATION METHODS ON BIOASSAY RESULTS

    EPA Science Inventory

    Four bioassays [Microtax(tm), Mutatox(tm), sister chromatid exchange (SCE), and metabolic cooperation] were used to analyze marine sediment extracts fractionated by two different methods: silica gel column chromatography and acid-base fractionation. esults indicated that a sedime...

  20. Statistical considerations in the analysis of data from replicated bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Multiple-dose bioassay is generally the preferred method for characterizing virulence of insect pathogens. Linear regression of probit mortality on log dose enables estimation of LD50/LC50 and slope, the latter having substantial effect on LD90/95s (doses of considerable interest in pest management)...

  1. US Army Radiological Bioassay and Dosimetry: The RBD software package

    SciTech Connect

    Eckerman, K. F.; Ward, R. C.; Maddox, L. B.

    1993-01-01

    The RBD (Radiological Bioassay and Dosimetry) software package was developed for the U. S. Army Material Command, Arlington, Virginia, to demonstrate compliance with the radiation protection guidance 10 CFR Part 20 (ref. 1). Designed to be run interactively on an IBM-compatible personal computer, RBD consists of a data base module to manage bioassay data and a computational module that incorporates algorithms for estimating radionuclide intake from either acute or chronic exposures based on measurement of the worker's rate of excretion of the radionuclide or the retained activity in the body. In estimating the intake,RBD uses a separate file for each radionuclide containing parametric representations of the retention and excretion functions. These files also contain dose-per-unit-intake coefficients used to compute the committed dose equivalent. For a given nuclide, if measurements exist for more than one type of assay, an auxiliary module, REPORT, estimates the intake by applying weights assigned in the nuclide file for each assay. Bioassay data and computed results (estimates of intake and committed dose equivalent) are stored in separate data bases, and the bioassay measurements used to compute a given result can be identified. The REPORT module creates a file containing committed effective dose equivalent for each individual that can be combined with the individual's external exposure.

  2. Book Review: Bioassays with Arthropods: 2nd Edition

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The technical book "Bioassays with Arthropods: 2nd Edition" (2007. Jacqueline L. Robertson, Robert M. Russell, Haiganoush K, Preisler and N. E. Nevin, Eds. CRC Press, Boca Raton, FL, 224 pp.) was reviewed for the scientific readership of the peer-reviewed publication Journal of Economic Entomology. ...

  3. US Army Radiological Bioassay and Dosimetry: The RBD software package

    SciTech Connect

    Eckerman, K.F.; Ward, R.C.; Maddox, L.B.

    1993-01-01

    The RBD (Radiological Bioassay and Dosimetry) software package was developed for the U. S. Army Material Command, Arlington, Virginia, to demonstrate compliance with the radiation protection guidance 10 CFR Part 20 (ref. 1). Designed to be run interactively on an IBM-compatible personal computer, RBD consists of a data base module to manage bioassay data and a computational module that incorporates algorithms for estimating radionuclide intake from either acute or chronic exposures based on measurement of the worker`s rate of excretion of the radionuclide or the retained activity in the body. In estimating the intake,RBD uses a separate file for each radionuclide containing parametric representations of the retention and excretion functions. These files also contain dose-per-unit-intake coefficients used to compute the committed dose equivalent. For a given nuclide, if measurements exist for more than one type of assay, an auxiliary module, REPORT, estimates the intake by applying weights assigned in the nuclide file for each assay. Bioassay data and computed results (estimates of intake and committed dose equivalent) are stored in separate data bases, and the bioassay measurements used to compute a given result can be identified. The REPORT module creates a file containing committed effective dose equivalent for each individual that can be combined with the individual`s external exposure.

  4. BENTHIC INVERTEBRATE BIOASSAYS WITH TOXIC SEDIMENT AND PORE WATER

    EPA Science Inventory

    The relative sensitivities of bioassays to determine the toxicity of sediments were investigated and three methods of making the sample dilutions required to generate dose-response relationships were compared. he assays studied were: (a) Microtox, a 15-min assay of Photobacterium...

  5. Shape-encoded silica microparticles for multiplexed bioassays.

    PubMed

    Kim, Lily Nari; Kim, Mira; Jung, Keumsim; Bae, Hyung Jong; Jang, Jisung; Jung, Yushin; Kim, Jiyun; Kwon, Sunghoon

    2015-08-01

    Shape-encoded silica microparticles for use in multiplexed bioassays were fabricated by using optofluidic maskless lithography (OFML) and tetraethylorthosilicate (TEOS) polymerization. These encoded silica microparticles exhibit excellent bioconjugation properties and negligible non-specific analyte adsorption. Encoded silica microparticles could be useful in a wide variety of applications, including DNA- and protein-based diagnostics. PMID:26125980

  6. Microplate Bioassay for Determining Substrate Selectivity of "Candida rugosa" Lipase

    ERIC Educational Resources Information Center

    Wang, Shi-zhen; Fang, Bai-shan

    2012-01-01

    Substrate selectivity of "Candida rugosa" lipase was tested using "p"-nitrophenyl esters of increasing chain length (C[subscript 1], C[subscript 7], C[subscript 15]) using the high-throughput screening method. A fast and easy 96-well microplate bioassay was developed to help students learn and practice biotechnological specificity screen. The…

  7. Sensitive bioassay for detection of biologically active ricin in food

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The potential use of ricin as an agent of biological warfare highlights the need to develop fast and effective methods to detect biologically active ricin. The current “gold standard” for ricin detection is an in vivo mouse bioassay; however, this method is not practical to test on a large number of...

  8. Correction of spray concentration and bioassay cage penetration data

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Field trials were conducted to demonstrate the need for correcting sampled spray concentration data for sampler collection efficiencies and estimated spray exposure levels in mosquito bioassays for cage interference effects. A large spray block was targeted with aerial spray treatments of etofenpro...

  9. 1. VIEW IN ROOM 125, BIOASSAY LABORATORY, SHOWN IS THE ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    1. VIEW IN ROOM 125, BIOASSAY LABORATORY, SHOWN IS THE FIRST STEP IN A SIX-STEP PROCESS TO ANALYZE URINE SAMPLES FOR PLUTONIUM AND URANIUM CONTAMINATION. IN THIS STEP, NITRIC ACID IS ADDED TO SAMPLE, AND THE SAMPLE IS BOILED DOWN TO A WHITE POWDER. - Rocky Flats Plant, Health Physics Laboratory, On Central Avenue between Third & Fourth Streets, Golden, Jefferson County, CO

  10. Sensitive bioassay for detection of PPARα potentially hazardous ligands with gold nanoparticle probe.

    PubMed

    Xia, Wei; Wan, Yan-Jian; Wang, Xianliang; Li, Yuan-Yuan; Yang, Wen-Jie; Wang, Chun-Xiang; Xu, Shun-qing

    2011-09-15

    There are so many kinds of peroxisome proliferator-activated receptor α (PPARα) ligands with hazardous effect for human health in the environment, such as certain herbicides, plasticizers and drugs. Among these agonists, perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), and mono-(2-ethylhexyl) phthalate (MEHP) are mostly investigated due to their persistence and accumulation in environment and their potential toxicity via PPARα. This investigation aims at developing a bioassay method to detect PPARα ligands based on the ligand-receptor interaction on microplate. PPARα, which formed heterodimers with retinoid X receptor-α (RXRα), were activated by PPARα ligands to form ligands-PPARα-RXRα complexes. Then the complexes were transferred into a microplate and captured via monoclonal anti-PPARα antibody. The PPARα responsive elements (PPRE) modified-gold nanoparticle probes were captured by the ligand-PPARα-RXRα complexes immobilized on the microplate, and then could be quantified through measuring the optical density after silver enhancement. The results showed that PFOS was quantified with a linear range from 100 pM to 1 μM and the detection limit was 10 pM. In addition to PFOS, PFOA and MEHP were also quantified within a proper range through the proposed bioassay. This bioassay was compared with that of liquid chromatography tandem-mass spectrometry (LC-MS) for water spiked samples with a significant correlation (r = 0.9893). This study provides a high-throughput detection method for PPARα ligands in microplate with high sensitivity and wide linear range. It may serve as an assistant of LC-MS for prescreening of PPARα ligands like PFOS. PMID:21726938

  11. Development of a Cell-Based Bioassay for Phospholipase A2-Triggered Liposomal Drug Release

    PubMed Central

    Arouri, Ahmad; Trojnar, Jakub; Schmidt, Steffen; Hansen, Anders H.; Mollenhauer, Jan; Mouritsen, Ole G.

    2015-01-01

    The feasibility of exploiting secretory phospholipase A2 (sPLA2) enzymes, which are overexpressed in tumors, to activate drug release from liposomes precisely at the tumor site has been demonstrated before. Although the efficacy of the developed formulations was evaluated using in vitro and in vivo models, the pattern of sPLA2-assisted drug release is unknown due to the lack of a suitable bio-relevant model. We report here on the development of a novel bioluminescence living-cell-based luciferase assay for the monitoring of sPLA2-triggered release of luciferin from liposomes. To this end, we engineered breast cancer cells to produce both luciferase and sPLA2 enzymes, where the latter is secreted to the extracellular medium. We report on setting up a robust and reproducible bioassay for testing sPLA2-sensitive, luciferin remote-loaded liposomal formulations, using 1,2-distearoyl-sn-glycero-3-phosphatidylcholine/1,2-distearoyl-sn-glycero-3-phosphatidylglycerol (DSPC/DSPG) 7:3 and DSPC/DSPG/cholesterol 4:3:3 as initial test systems. Upon their addition to the cells, the liposomes were degraded almost instantaneously by sPLA2 releasing the encapsulated luciferin, which provided readout from the luciferase-expressing cells. Cholesterol enhanced the integrity of the formulation without affecting its susceptibility to sPLA2. PEGylation of the liposomes only moderately broadened the release profile of luciferin. The provided bioassay represents a useful tool for monitoring active drug release in situ in real time as well as for testing and optimizing of sPLA2-sensitive lipid formulations. In addition, the bioassay will pave the way for future in-depth in vitro and in vivo studies. PMID:25945937

  12. Genotoxicity of leachates from a landfill using three bioassays.

    PubMed

    Cabrera, G L; Rodriguez, D M

    1999-05-19

    In the city of Queretaro, around 500 tons of solid wastes are produced everyday and are deposited in a landfill. This is the result of social and economic activities of human beings or from their normal physiological functions. As a result of rain, leachates are produced, which, if not handled and treated correctly, may pollute the underground water. Among the bioassays developed for the detection of mutagenicity in environmental pollutants, plant systems have been proven to be sensitive, cheap, and effective. The purpose of this study was to determine the presence of genotoxic agents in the leachates of the landfill of the city using three bioassays: Tradescantia-micronucleus (Trad-MCN), Tradescantia stamen hair mutations (Trad-SHM) and Allium root anaphase aberrations (AL-RAA) and make a comparison of the results in the three assays. Leachates were sampled during both the dry and rainy seasons. Plant cuttings of Tradescantia or the roots of Allium were treated by submerging them in the leachates. Three replicates of each sample were analyzed in each of the three bioassays. As expected the samples of leachates collected during the dry season showed a higher genotoxicity than those collected during the rainy season. In conclusion, there are substances present in the leachates capable of inducing genotoxicity in the plant assays. On the other hand, the plant assays showed different degrees of sensitivity: the more sensitive was the Trad-MCN bioassay and the less sensitive the Trad-SHM assay. Therefore, when analyzing environmental pollutants it is recommended to use a battery of bioassays. PMID:10350599

  13. Immobilized magnetic beads based multi-target affinity selection coupled with high performance liquid chromatography-mass spectrometry for screening anti-diabetic compounds from a Chinese medicine "Tang-Zhi-Qing".

    PubMed

    Tao, Yi; Chen, Zhui; Zhang, Yufeng; Wang, Yi; Cheng, Yiyu

    2013-05-01

    We developed an approach for screening bioactive compounds from botanical drug using multiple target-immobilized magnetic beads coupled with high performance liquid chromatography-mass spectrometry. This novel approach was called magnetic beads based multi-target affinity selection-mass spectrometry (MT-ASMS). It can enrich and identify different types of ligands from mixture extracts. Multiple targets (maltase, invertase, lipase) were immobilized on the magnetic beads by covalent linkage using 1-(3-dimethyl-aminopropyl)-3-ethyl-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) as reaction reagents, respectively. The properties of enzyme conjugated magnetic beads were characterized using transmission electron microscopy, X-ray diffractometer and vibration sample magnetometer. Several factors including pH, ion strength, incubation time and temperature were optimized using three known ligands (caffeic acid, ferulic acid, and hesperidin). The established MT-ASMS approach was applied to screening for ligands from a Chinese medicine "Tang-Zhi-Qing", which was used to treat type II diabetes in China. Seven bound compounds were identified via liquid chromatography-mass spectrometry (LC/MS). Five active compounds including 2,3,4,6-tetra-O-galloyl-D-glucose, 1,2,3,4-tetra-O-galloyl-D-glucose, 1,2,3,4,6-penta-O-galloyl-d-glucose, quercetin-3-O-β-D-glucuronide and quercetin-3-O-β-D-glucoside were identified and their activities were validated by conventional inhibitory assay. Our findings suggested that the proposed approach is efficient in screening compounds with multiple activities from extracts of botanical drugs. PMID:23501439

  14. BioAssay Ontology (BAO): a semantic description of bioassays and high-throughput screening results

    PubMed Central

    2011-01-01

    Background High-throughput screening (HTS) is one of the main strategies to identify novel entry points for the development of small molecule chemical probes and drugs and is now commonly accessible to public sector research. Large amounts of data generated in HTS campaigns are submitted to public repositories such as PubChem, which is growing at an exponential rate. The diversity and quantity of available HTS assays and screening results pose enormous challenges to organizing, standardizing, integrating, and analyzing the datasets and thus to maximize the scientific and ultimately the public health impact of the huge investments made to implement public sector HTS capabilities. Novel approaches to organize, standardize and access HTS data are required to address these challenges. Results We developed the first ontology to describe HTS experiments and screening results using expressive description logic. The BioAssay Ontology (BAO) serves as a foundation for the standardization of HTS assays and data and as a semantic knowledge model. In this paper we show important examples of formalizing HTS domain knowledge and we point out the advantages of this approach. The ontology is available online at the NCBO bioportal http://bioportal.bioontology.org/ontologies/44531. Conclusions After a large manual curation effort, we loaded BAO-mapped data triples into a RDF database store and used a reasoner in several case studies to demonstrate the benefits of formalized domain knowledge representation in BAO. The examples illustrate semantic querying capabilities where BAO enables the retrieval of inferred search results that are relevant to a given query, but are not explicitly defined. BAO thus opens new functionality for annotating, querying, and analyzing HTS datasets and the potential for discovering new knowledge by means of inference. PMID:21702939

  15. Isolation of Fungi from Heterodera glycines and in vitro Bioassays for Their Antagonism to Eggs.

    PubMed

    Meyer, S L; Huettel, R N; Sayre, R M

    1990-10-01

    Twenty fungi were assayed in vitro for antagonism to eggs of Heterodera glycines. Eight of the fungi were isolated from cysts or eggs of H. glycines during the current study, one was isolated from Panagrellus redivivus, and eleven were obtained from other researchers or collections. The bioassays were conducted on eggs from nematodes that had been grown monoxenically on excised root tips. Phoma chrysanthemicola, one strain of Verticillium chlamydosporium, and one strain of V. lecanii caused a decrease (P < 0.01, P < 0.05, P < 0.05, respectively) in the number of viable eggs, although no hyphae were observed colonizing live eggs. Trichoderma polysporum infected live eggs but enhanced (P < 0.05) egg survival. Acremonium bacillisporum, Chaetomium sp., Drechmeria coniospora (two strains), Epicoccum sp., Exophiala jeanselmei, Fusarium sp., Neocosmospora vasinfecta, Scytalidium fulvum, Trichoderma harzianum (two strains), V. chlamydosporium (one strain), V. lecanii (three strains), and an unidentified fungus did not measurably affect egg viability, even though hyphae of five of these fungi were seen in live eggs. The bioassay provides a useful step in the selection of a biological control agent for this major nematode pest. PMID:19287754

  16. Pullulan encapsulation of labile biomolecules to give stable bioassay tablets.

    PubMed

    Jahanshahi-Anbuhi, Sana; Pennings, Kevin; Leung, Vincent; Liu, Meng; Carrasquilla, Carmen; Kannan, Balamurali; Li, Yingfu; Pelton, Robert; Brennan, John D; Filipe, Carlos D M

    2014-06-10

    A simple and inexpensive method is reported for the long-term stabilization of enzymes and other unstable reagents in premeasured quantities in water-soluble tablets (cast, not compressed) made with pullulan, a nonionic polysaccharide that forms an oxygen impermeable solid upon drying. The pullulan tablets dissolve in aqueous solutions in seconds, thereby facilitating the easy execution of bioassays at remote sites with no need for special reagent handling and liquid pipetting. This approach is modular in nature, thus allowing the creation of individual tablets for enzymes and their substrates. Proof-of-principle demonstrations include a Taq polymerase tablet for DNA amplification through PCR and a pesticide assay kit consisting of separate tablets for acetylcholinesterase and its chromogenic substrate, indoxyl acetate, both of which are highly unstable. The encapsulated reagents remain stable at room temperature for months, thus enabling the room-temperature shipping and storage of bioassay components. PMID:24764260

  17. A New Bioassay for Auxins and Cytokinins 1

    PubMed Central

    Boerjan, Wout; Genetello, Chris; Van Montagu, Marc; Inzé, Dirk

    1992-01-01

    The authors have developed a sensitive bioassay that can be used to detect auxins as well as cytokinins. The bioassay is based on the expression in transformed tobacco (Nicotiana tabacum) mesophyll protoplasts of a chimeric gene, consisting of the upstream sequences of the Agrobacterium tumefaciens gene 5, coupled to the coding sequence of the β-glucuronidase. The expression of this gene is induced by the presence of both auxin and cytokinin in the culture medium. Using this assay, indole-3-acetic acid was detected at 5 × 10−8 molar, whereas trans-zeatin could be detected at 5 × 10−11 molar. The assay can be performed in microtiter plates, allowing numerous samples to be analyzed simultaneously. Only 2.5 × 105 protoplasts are required for one individual assay in 250 microliters of culture medium and for qualitative results, the reaction is readily visualized by ultraviolet light. ImagesFigure 3Figure 4Figure 6 PMID:16668975

  18. A Rapid and Simple Bioassay Method for Herbicide Detection

    PubMed Central

    Li, Xiu-Qing; Ng, Alan; King, Russell; Durnford, Dion G.

    2008-01-01

    Chlamydomonas reinhardtii, a unicellular green alga, has been used in bioassay detection of a variety of toxic compounds such as pesticides and toxic metals, but mainly using liquid culture systems. In this study, an algal lawn—agar system for semi-quantitative bioassay of herbicidal activities has been developed. Sixteen different herbicides belonging to 11 different categories were applied to paper disks and placed on green alga lawns in Petri dishes. Presence of herbicide activities was indicated by clearing zones around the paper disks on the lawn 2–3 days after application. The different groups of herbicides induced clearing zones of variable size that depended on the amount, mode of action, and chemical properties of the herbicides applied to the paper disks. This simple, paper-disk-algal system may be used to detect the presence of herbicides in water samples and act as a quick and inexpensive semi-quantitative screening for assessing herbicide contamination. PMID:19578512

  19. Paper bioassay based on ceria nanoparticles as colorimetric probes.

    PubMed

    Ornatska, Maryna; Sharpe, Erica; Andreescu, Daniel; Andreescu, Silvana

    2011-06-01

    We report the first use of redox nanoparticles of cerium oxide as colorimetric probes in bioanalysis. The method is based on changes in the physicochemical properties of ceria nanoparticles, used here as chromogenic indicators, in response to the analyte. We show that these particles can be fully integrated in a paper-based bioassay. To construct the sensor, ceria nanoparticles and glucose oxidase were coimmobilized onto filter paper using a silanization procedure. In the presence of glucose, the enzymatically generated hydrogen peroxide induces a visual color change of the ceria nanoparticles immobilized onto the bioactive sensing paper, from white-yellowish to dark orange, in a concentration-dependent manner. A detection limit of 0.5 mM glucose with a linear range up to 100 mM and a reproducibility of 4.3% for n = 11 ceria paper strips were obtained. The assay is fully reversible and can be reused for at least 10 consecutive measurement cycles, without significant loss of activity. Another unique feature is that it does not require external reagents, as all the sensing components are fixed onto the paper platform. The bioassay can be stored for at least 79 days at room temperature while maintaining the same analytical performance. An example of analytical application was demonstrated for the detection of glucose in human serum. The results demonstrate the potential of this type of nanoparticles as novel components in the development of robust colorimetric bioassays. PMID:21524141

  20. A Bioassay System Using Bioelectric Signals from Small Fish

    NASA Astrophysics Data System (ADS)

    Terawaki, Mitsuru; Soh, Zu; Hirano, Akira; Tsuji, Toshio

    Although the quality of tap water is generally examined using chemical assay, this method cannot be used for examination in real time. Against such a background, the technique of fish bioassay has attracted attention as an approach that enables constant monitoring of aquatic contamination. The respiratory rhythms of fish are considered an efficient indicator for the ongoing assessment of water quality, since they are sensitive to chemicals and can be indirectly measured from bioelectric signals generated by breathing. In order to judge aquatic contamination accurately, it is necessary to measure bioelectric signals from fish swimming freely as well as to stably discriminate measured signals, which vary between individuals. However, no bioassay system meeting the above requirements has yet been established. This paper proposes a bioassay system using bioelectric signals generated from small fish in free-swimming conditions. The system records signals using multiple electrodes to cover the extensive measurement range required in a free-swimming environment, and automatically discriminates changes in water quality from signal frequency components. This discrimination is achieved through an ensemble classification method using probability neural networks to solve the problem of differences between individual fish. The paper also reports on the results of related validation experiments, which showed that the proposed system was able to stably discriminate between water conditions before and after bleach exposure.

  1. Modeling development of inhibition zones in an agar diffusion bioassay.

    PubMed

    Chandrasekar, Vaishnavi; Knabel, Stephen J; Anantheswaran, Ramaswamy C

    2015-09-01

    A two-temperature agar diffusion bioassay is commonly used to quantify the concentration of nisin using Micrococcus luteus as the indicator microorganism. A finite element computational model based on Fick's second law of diffusion was used to predict the radius of the inhibition zone in this diffusion bioassay. The model developed was used to calculate nisin concentration profiles as a function of time and position within the agar. The minimum inhibitory concentration (MIC) of nisin against M. luteus was determined experimentally. The critical time (T c) for growth of M. luteus within the agar diffusion bioassay was experimentally determined using incubation studies with nisin. The radius of the inhibition zone was predicted from the computational model as the location where the predicted nisin concentration at T c was equal to MIC. The MIC was experimentally determined to be 0.156 μg mL(-1), and T c was determined to be 7 h. Good agreement (R (2) = 0.984) was obtained between model-predicted and experimentally determined inhibition zone radii. PMID:26405525

  2. Improved bioassay for detecting autoinducer of Rhodovulum sulfidophilum

    NASA Astrophysics Data System (ADS)

    Terada, T.; Kikuchi, Y.; Umekage, S.

    2015-02-01

    Quorum sensing is a bacterial gene regulation system that enables prompt environmental adaptation in response to cell density. Quorum sensing is driven by an extracellularly secreted chemical signal called autoinducer. Gram-negative bacteria produce one or several types of N-acylhomoserine lactone (AHL) as autoinducers. Our previous study suggests that the gram-negative marine photosynthetic bacterium Rhodovulum sulfidophilum produces AHL in the early stationary phase and plays a role in maintaining the bacterial cell aggregates called "floc". We performed conventional bioassay to identify AHL production by using Chromobacterium violaceum VIR07, which produces violet pigment (violacein) in response to AHL with side chains ranging from C10 to C18 in length. However, we were not able to observe the violacein with good reproducibility, suggesting that inhibitory chemical compounds co-existed in the AHL extract. Therefore, we improved the extraction method; the ethyl acetate-extracted AHLs were fractionated by using reverse phase TLC. By using the re-extracted AHLs for the bioassay, we observed an obvious production of violacein. This result clearly indicates that R. sulfidophilum produces AHLs with side chains ranging from C10 to C18 in length and suggests the utility of improved bioassay for AHL detection.

  3. Modeling development of inhibition zones in an agar diffusion bioassay

    PubMed Central

    Chandrasekar, Vaishnavi; Knabel, Stephen J; Anantheswaran, Ramaswamy C

    2015-01-01

    A two-temperature agar diffusion bioassay is commonly used to quantify the concentration of nisin using Micrococcus luteus as the indicator microorganism. A finite element computational model based on Fick's second law of diffusion was used to predict the radius of the inhibition zone in this diffusion bioassay. The model developed was used to calculate nisin concentration profiles as a function of time and position within the agar. The minimum inhibitory concentration (MIC) of nisin against M. luteus was determined experimentally. The critical time (Tc) for growth of M. luteus within the agar diffusion bioassay was experimentally determined using incubation studies with nisin. The radius of the inhibition zone was predicted from the computational model as the location where the predicted nisin concentration at Tc was equal to MIC. The MIC was experimentally determined to be 0.156 μg mL−1, and Tc was determined to be 7 h. Good agreement (R2 = 0.984) was obtained between model-predicted and experimentally determined inhibition zone radii. PMID:26405525

  4. Novel bioassay using Bacillus megaterium to detect tetracycline in milk.

    PubMed

    Tumini, Melisa; Nagel, Orlando G; Molina, Pilar; Althaus, Rafael L

    2016-01-01

    Tetracyclines are used for the prevention and control of dairy cattle diseases. Residues of these drugs can be excreted into milk. Thus, the aim of this study was to develop a microbiological method using Bacillus megaterium to detect tetracyclines (chlortetracycline, oxytetracycline and tetracycline) in milk. In order to approximate the limits of detection of the bioassay to the Maximum Residue Limit (100μg/l) for milk tetracycline, different concentrations of chloramphenicol (0, 1000, 1500 and 2000μg/l) were tested. The detection limits calculated were similar to the Maximum Residue Limits when a bioassay using B. megaterium ATCC 9885 spores (2.8×10(8)spores/ml) and chloramphenicol (2000μg/l) was utilized. This bioassay detects 105μg/l of chlortetracycline, 100μg/l of oxytetracycline and 134μg/l of tetracycline in 5h. Therefore, this method is suitable to be incorporated into a microbiological multi-residue system for the identification of tetracyclines in milk. PMID:27131738

  5. Nanoparticle-catalyzed reductive bleaching for fabricating turn-off and enzyme-free amplified colorimetric bioassays.

    PubMed

    Li, Wei; Qiang, Weibing; Li, Jie; Li, Hui; Dong, Yifan; Zhao, Yaju; Xu, Danke

    2014-01-15

    Nanoparticle-catalyzed reductive bleaching reactions of colored substrates are emerging as a class of novel indicator reactions for fabricating enzyme-free amplified colorimetric biosensing (turn-off mode), which are exactly opposite to the commonly used oxidative coloring processes of colorless substrates in traditional enzyme-catalyzed amplified colorimetric bioassays (turn-on mode). In this work, a simple theoretical analysis shows that the sensitivity of this colorimetric bioassay can be improved by increasing the amplification factor (kcatΔt), or enhancing the binding affinity between analyte and receptor (Kd), or selecting the colored substrates with high extinction coefficients (ε). Based on this novel strategy, we have developed a turn-off and cost-effective amplified colorimetric thrombin aptasensor. This aptasensor made full use of sandwich binding of two affinity aptamers for increased specificity, magnetic particles for easy separation and enrichment, and gold nanoparticle (AuNP)-catalyzed reductive bleaching reaction to generate the amplified colorimetric signal. With 4-nitrophenol (4-NP) as the non-dye colored substrate, colorimetric bioassay of thrombin was achieved by the endpoint method with a detection limit of 91pM. In particular, when using methylene blue (MB) as the substrate, for the first time, a more convenient and efficient kinetic-based colorimetric thrombin bioassay was achieved without the steps of acidification termination and magnetic removal of particles, with a low detection limit of 10pM, which was superior to the majority of the existing colorimetric thrombin aptasensors. The proposed colorimetric protocol is expected to hold great promise in field analysis and point-of-care applications. PMID:23962710

  6. Toxicity of copper-spiked sediments to Tubifex tubifex (Oligochaeta, Tubificidae): Comparison of the 28-day reproductive bioassay with an early-life-stage bioassay

    SciTech Connect

    Vecchi, M.; Pasteris, A.; Bonomi, G. . Dipt. di Biologia Evoluzionistica Sperimentale); Reynoldson, T.B. . National Water Research Inst.)

    1999-06-01

    Two sediment bioassay methods using Tubifex tubifex (Mueller, 1774) as the test species were compared. The first was an adult reproduction test, the second an early-life-stage survival test. The duration of both bioassays is 28 d and the amount of work required was similar; they may be useful alternatives to each other in different circumstances (e.g., the early life stage bioassay could be carried out with smaller volumes of sediment). The two bioassays were performed simultaneously on copper-spiked sediments. Sediments from two freshwater and two terrestrial sites were used; five separate, nonsimultaneous experiments were performed, one for each sediment or soil and a further experiment with soil with a good supplement. In the adult bioassay, there were large differences in the production of cocoons, eggs, and young among the control treatments of the five experiments. There were also major differences in the NOEC and LOEC for copper between the tested substrates. The early life stage bioassay appears to be less sensitive to copper toxicity than the adult reproductive bioassay since NOECs and LOECs are higher for early survival than for the most sensitive endpoints of the adult bioassay in three experiments out of five.

  7. Toxicity assessment of effluent from flash light manufacturing industry by bioassay tests in Trigonella foenumgracum.

    PubMed

    Kumari, Narendra; Kumar, Sanjeev; Bauddh, Kuldeep; Dwivedi, Neetu; Singh, D P; Barman, S C

    2014-11-01

    A rapid bioassay test was conducted to study heavy metal accumulation and biochemical changes in Trigonella foenumgracum (methi) irrigated with 25, 50, 75 and 100% of effluent from flash light manufacturing industry at 60 days after sowing. Total metal concentration in effluent samples was: Cr = 0.12 < Cd = 0.18 < Pb = 0.24 < Cu = 2.68 mg l(-1) whereas, metals were not detected in control. An increase in photosynthetic pigments of exposed plant was noticed up to 50% concentrations of the effluent followed by a decrease at higher concentration as compared to their respective control.An enhanced lipid peroxidation in the treated plants was observed, which was evident by increased level of antioxidants: proline, cysteine, malondialdehyde and ascorbic acid content. The treated plants accumulated metals in the following order: Cu > Pb > Cr > Cd in the roots and shoots. PMID:25522513

  8. Microwave-accelerated bioassay technique for rapid and quantitative detection of biological and environmental samples.

    PubMed

    Mohammed, Muzaffer; Syed, Maleeha F; Aslan, Kadir

    2016-01-15

    Quantitative detection of molecules of interest from biological and environmental samples in a rapid manner, particularly with a relevant concentration range, is imperative to the timely assessment of human diseases and environmental issues. In this work, we employed the microwave-accelerated bioassay (MAB) technique, which is based on the combined use of circular bioassay platforms and microwave heating, for rapid and quantitative detection of Glial Fibrillary Acidic Protein (GFAP) and Shiga like toxin (STX 1). The proof-of-principle use of the MAB technique with the circular bioassay platforms for the rapid detection of GFAP in buffer based on colorimetric and fluorescence readouts was demonstrated with a 900W kitchen microwave. We also employed the MAB technique with a new microwave system (called the iCrystal system) for the detection of GFAP from mice with brain injuries and STX 1 from a city water stream. Control bioassays included the commercially available gold standard bioassay kits run at room temperature. Our results show that the lower limit of detection (LLOD) of the colorimetric and fluorescence based bioassays for GFAP was decreased by ~1000 times using the MAB technique and our circular bioassay platforms as compared to the commercially available bioassay kits. The overall bioassay time for GFAP and STX 1 was reduced from 4h using commercially available bioassay kits to 10min using the MAB technique. PMID:26356762

  9. New technique for collecting ambient diesel particles for bioassays

    SciTech Connect

    Hallock, M.F.; Smith, T.J.; Hammond, S.K.; Beck, B.D.; Brain, J.D.

    1987-05-01

    This paper describes a new application of viable aerosol sampler, the Liquid electrostatic Aerosol Precipitator (LEAP), for the collection of diesel particles for bioassays of pulmonary toxicity and mutagenicity or carinogenicity. Currently used methods (filtration, dry electrostatic precipitation) cause agglomeration of particles and increases in particle size up to twenty-fold, which may alter particle toxicity significantly. Collection of diesel particles with the LEAP preserved submicronic particle size. Differences in chemical composition of extracts of surface adsorbents as compared to particles collected on filters also were observed. This technique may be applicable for collection other types of combustion products or oil mists that agglomerate when collected by filtration.

  10. A new technique for collecting ambient diesel particles for bioassays.

    PubMed

    Hallock, M F; Smith, T J; Hammond, S K; Beck, B D; Brain, J D

    1987-05-01

    This paper describes a new application of a viable aerosol sampler, the Liquid Electrostatic Aerosol Precipitator (LEAP), for the collection of diesel particles for bioassays of pulmonary toxicity and mutagenicity or carcinogenicity. Currently used methods (filtration, dry electrostatic precipitation) cause agglomeration of particles and increases in particle size up to twenty-fold, which may alter particle toxicity significantly. Collection of diesel particles with the LEAP preserved submicronic particle size. Differences in chemical composition of extracts of surface adsorbents as compared to particles collected on filters also were observed. This technique may be applicable for collection of other types of combustion products or oil mists that agglomerate when collected by filtration. PMID:2438921

  11. Field and Bioassay Indicators for Internal Dose Intervention Therapy

    SciTech Connect

    Carbaugh, Eugene H.

    2007-05-01

    Guidance is presented that is used at the U.S. Department of Energy Hanford Site to identify the potential need for medical intervention in response to intakes of radioactivity. The guidance, based on ICRP Publication 30 models and committed effective dose equivalents of 20 mSv and 200 mSv, is expressed as numerical workplace measurements and derived first-day bioassay results for large intakes. It is used by facility radiation protection staff and on-call dosimetry support staff during the first few days following an intake.

  12. Field and bioassay indicators for internal dose intervention therapy.

    PubMed

    Carbaugh, Eugene H

    2007-05-01

    Guidance is presented that is used at the U.S. Department of Energy Hanford Site to identify the potential need for medical intervention in response to intakes of radioactivity. The guidance, based on ICRP Publication 30 models and committed effective dose equivalents of 20 mSv and 200 mSv, is expressed as numerical workplace measurements and derived first-day bioassay results for large intakes. It is used by facility radiation protection staff and on-call dosimetry support staff during the first few days following an intake. PMID:17440323

  13. Electroantennographic bioassay as a screening tool for host plant volatiles.

    PubMed

    Beck, John J; Light, Douglas M; Gee, Wai S

    2012-01-01

    Plant volatiles play an important role in plant-insect interactions. Herbivorous insects use plant volatiles, known as kairomones, to locate their host plant. When a host plant is an important agronomic commodity feeding damage by insect pests can inflict serious economic losses to growers. Accordingly, kairomones can be used as attractants to lure or confuse these insects and, thus, offer an environmentally friendly alternative to pesticides for insect control. Unfortunately, plants can emit a vast number volatiles with varying compositions and ratios of emissions dependent upon the phenology of the commodity or the time of day. This makes identification of biologically active components or blends of volatile components an arduous process. To help identify the bioactive components of host plant volatile emissions we employ the laboratory-based screening bioassay electroantennography (EAG). EAG is an effective tool to evaluate and record electrophysiologically the olfactory responses of an insect via their antennal receptors. The EAG screening process can help reduce the number of volatiles tested to identify promising bioactive components. However, EAG bioassays only provide information about activation of receptors. It does not provide information about the type of insect behavior the compound elicits; which could be as an attractant, repellent or other type of behavioral response. Volatiles eliciting a significant response by EAG, relative to an appropriate positive control, are typically taken on to further testing of behavioral responses of the insect pest. The experimental design presented will detail the methodology employed to screen almond-based host plant volatiles by measurement of the electrophysiological antennal responses of an adult insect pest navel orangeworm (Amyelois transitella) to single components and simple blends of components via EAG bioassay. The method utilizes two excised antennae placed across a "fork" electrode holder. The protocol

  14. An emergency bioassay method for (210)Po in urine.

    PubMed

    Guérin, Nicolas; Dai, Xiongxin

    2015-09-01

    A rapid method was developed to efficiently measure (210)Po in urine samples in an emergency situation. Polonium-210 in small urine samples (10 mL) was spontaneously deposited on a stainless steel disc in 1 M HCl at room temperature for 4 h in a polyethylene bottle. The metallic disc was then counted for 4 h by alpha spectrometry. The developed method allowed the preparation of large sample batch in a short time. The method meets the requirements for an emergency bioassay procedure. PMID:26115206

  15. How to Fabricate Functional Artificial Luciferases for Bioassays.

    PubMed

    Kim, Sung-Bae; Fujii, Rika

    2016-01-01

    The present protocol introduces fabrication of artificial luciferases (ALuc(®)) by extracting the consensus amino acids from the alignment of copepod luciferase sequences. The made ALucs have unique sequential identities that are phylogenetically distinctive from those of any existing copepod luciferase. Some ALucs exhibited heat stability, and strong and greatly prolonged optical intensities. The made ALucs are applicable to various bioassays as an optical readout, including live cell imaging, single-chain probes, and bioluminescent tags of antibodies. The present protocol guides on how to fabricate a unique artificial luciferase with designed optical properties and functionalities. PMID:27424894

  16. [Evaluation of Antilles fish ciguatoxicity by mouse and chick bioassays].

    PubMed

    Pottier, I; Vernoux, J P

    2003-03-01

    Ciguatera is a common seafood poisoning in Western Atlantic and French West Indies. Ciguatera fish poisoning in the Caribbean is a public health problem. A toxicological study was carried out on 178 Caribbean fish specimens (26 species) captured off Guadeloupe and Saint Barthelemy between 1993 and 1999. The mouse bioassay and the chick feeding test were used to control fish edibility. Ciguatoxins presence was assumed when symptomatology was typical of ciguatera in mouse and chick. Fishes were classified in three groups: non toxic fish (edible), low toxic fish (not edible) and toxic fish (not edible). 75% of fishes were non toxic. Toxic fish specimens belonged to four families of high trophic level carnivores: Carangidae, Lutjanidae, Serranidae et Sphyraenidae. Percentages of toxic fishes to humans reached 55% for Caranx latus and 33% for Caranx bartholomaei and Caranx lugubris. Only a significant correlation between weight and toxicity was only found for C. latus and snappers. Small carnivorous groupers (Serranidae) were also toxic. Atoxic fish species were (a) pelagic fish (Coryphaena hippurus, Auxis thazard and Euthynnus pelamis), (b) invertebrates feeders (Malacanthus plumieri, Balistes vetula), (c) small high-risk fish or (d) fish of edible benthic fish families. Liver of four fishes (Mycteroperca venenosa, Caranx bartholomaei, Seriola rivoliana, Gymnothorax funebris) contained ciguatoxins at a significant level although their flesh was safe. This study confirms the usefulness of mouse and chick bioassays for sanitary control of fish. PMID:12784589

  17. A Bioassay for Lafora Disease and Laforin Glucan Phosphatase Activity

    PubMed Central

    Sherwood, Amanda R.; Johnson, Mary Beth; Delgado-Escueta, Antonio V.; Gentry, Matthew S.

    2013-01-01

    Objectives Lafora disease is a rare yet invariably fatal form of progressive neurodegenerative epilepsy resulting from mutations in the phosphatase laforin. Several therapeutic options for Lafora disease patients are currently being explored, and these therapies would benefit from a biochemical means of assessing functional laforin activity following treatment. To date, only clinical outcomes such as decreases in seizure frequency and severity have been used to indicate success of epilepsy treatment. However, these qualitative measures exhibit variability and must be assessed over long periods of time. In this work, we detail a simple and sensitive bioassay that can be used for the detection of functional endogenous laforin from human and mouse tissue. Design and methods We generated antibodies capable of detecting and immunoprecipitating endogenous laforin. Following laforin immunoprecipitation, laforin activity was assessed via phosphatase assays using para-nitrophenylphosphate (pNPP) and a malachite green-based assay specific for glucan phosphatase activity. Results We found that antibody binding to laforin does not impede laforin activity. Furthermore, the malachite green-based glucan phosphatase assay used in conjunction with a rabbit polyclonal laforin antibody was capable of detecting endogenous laforin activity from human and mouse tissue. Importantly, this assay discriminated between laforin activity and other phosphatases. Conclusions The bioassay that we have developed utilizing laforin antibodies and an assay specific for glucan phosphatase activity could prove valuable in the rapid detection of functional laforin in patients to which novel Lafora disease therapies have been administered. PMID:24012855

  18. Dichloromethane attracts diabroticite larvae in a laboratory behavioral bioassay.

    PubMed

    Jewett, D K; Bjostad, L B

    1996-07-01

    A two-choice laboratory behavioral bioassay was used to demonstrate that dichloromethane elicits the dose-dependent attraction of secondinstar western and southern corn rootworms. Preliminary data suggest that second-instar banded cucumber beetles are also attracted to dichloromethane. An eluotropic series of 10 materials, including distilled water, ethanol, methanol, acetone, ethyl dichloroacetate, dichloromethane, diethyl ether, benzene, hexadecane, and hexane, was tested for attraction of western corn rootworm larvae. Dichloromethane was the only one attractive at all doses tested, and orthogonal comparisons revealed a quadratic trend (convex) for responses of larvae to increasing dose. Benzene and hexadecane also attracted larvae, but significantly fewer than dichloromethane, and only at three doses and one dose, respectively. Orthogonal comparisons revealed no linear or quadratic trend for responses of larvae to increasing doses of either compound. Dichloromethane is the first organic compound demonstrated to attract western corn rootworm larvae in the absence of carbon dioxide. Carbon dioxide has previously been reported to attract western corn rootworm larvae either independently or when combined with other organic compounds, and the sensitivity of our bioassay was tested by demonstrating the dose-dependent attraction of western corn rootworm larvae to carbonated water as a carbon dioxide source. We have also demonstrated the attraction of southern corn rootworm larvae to carbon dioxide and propose that carbon dioxide and dichloromethane behave analogously when they interact with chemoreceptor sites on larvae. PMID:24226089

  19. Vicia faba bioassay for environmental toxicity monitoring: A review.

    PubMed

    Iqbal, Munawar

    2016-02-01

    Higher plants are recognized as excellent genetic models to detect cytogenetic and mutagenic agents and are frequently used in environmental monitoring studies. Vicia faba (V. faba) bioassay have been used to study DNA damages i.e., chromosomal and nuclear aberrations induced by metallic compounds, pesticides, complex mixtures, petroleum derivates, toxins, nanoparticles and industrial effluents. The main advantages of using V. faba is its availability round the year, economical to use, easy to grow and handle; its use does not require sterile conditions, rate of cell division is fast, chromosomes are easy to score, less expensive and more sensitive as compared to other short-term tests that require pre-preparations. The V. faba test offers evaluation of different endpoints and tested agents can be classified as cytotoxic/genotoxic/mutagenic. This test also provides understanding about mechanism of action, whether the tested agent is clastogenic or aneugenic in nature. In view of advantages offered by V. faba test system, it is used extensively to assess toxic agents and has been emerged as an important bioassay for ecotoxicological studies. Based on the applications of V. faba test to assess the environmental quality, this article offers an overview of this test system and its efficiency in assessing the cytogenetic and mutagenic agents in different classes of the environmental concerns. PMID:26414739

  20. Resistance monitoring of Heliothis virescens to pyramided cotton varieties with a hydrateable, artificial cotton leaf bioassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Proof of concept was demonstrated for a practical, off the shelf bioassay to monitor for tobacco budworm resistance to pyramided Bt cotton using plant eluants. The bioassay was based on a previously described feeding disruption test using hydrateable artificial diet containing a blue indicator dye, ...

  1. Comparison of two mosquito bioassay methods for the estimate of minimum effective dose in repellents

    Technology Transfer Automated Retrieval System (TEKTRAN)

    It is expected that laboratory-based repellent bioassays should reliably evaluate the efficacy of compounds that deter mosquito feeding behavior. The variety of repellent bioassays available allows for flexibility in design, but makes it difficult to compare any two methods, including in vitro and i...

  2. Profiling Animal Toxicants by Automatically Mining Public Bioassay Data: A Big Data Approach for Computational Toxicology

    PubMed Central

    Zhang, Jun; Hsieh, Jui-Hua; Zhu, Hao

    2014-01-01

    In vitro bioassays have been developed and are currently being evaluated as potential alternatives to traditional animal toxicity models. Already, the progress of high throughput screening techniques has resulted in an enormous amount of publicly available bioassay data having been generated for a large collection of compounds. When a compound is tested using a collection of various bioassays, all the testing results can be considered as providing a unique bio-profile for this compound, which records the responses induced when the compound interacts with different cellular systems or biological targets. Profiling compounds of environmental or pharmaceutical interest using useful toxicity bioassay data is a promising method to study complex animal toxicity. In this study, we developed an automatic virtual profiling tool to evaluate potential animal toxicants. First, we automatically acquired all PubChem bioassay data for a set of 4,841 compounds with publicly available rat acute toxicity results. Next, we developed a scoring system to evaluate the relevance between these extracted bioassays and animal acute toxicity. Finally, the top ranked bioassays were selected to profile the compounds of interest. The resulting response profiles proved to be useful to prioritize untested compounds for their animal toxicity potentials and form a potential in vitro toxicity testing panel. The protocol developed in this study could be combined with structure-activity approaches and used to explore additional publicly available bioassay datasets for modeling a broader range of animal toxicities. PMID:24950175

  3. Effects of Wind Speed on Aerosol Spray Penetration in Adult Mosquito Bioassay Cages

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bioassay cages are commonly used to assess efficacy of insecticides against adult mosquitoes in the field. To properly correlate adult mortality readings to insecticidal efficacy and/or spray application parameters, it is important to know how the cage used in the bioassay interacts with the spray ...

  4. LIFE CYCLE BIOASSAY FOR ASSESSMENT OF THE EFFECTS OF TOXIC CHEMICALS USING RAPID CYCLING OF BRASSICA

    EPA Science Inventory

    Initial evaluation of a new plant life cycle bioassay for the assessment of the effects of toxic chemicals is presented. he bioassay features a rapid cycling Brassica species that can complete its life cycle in as little as 36 days. he herbicide dalapon (2,2 dichloropropionic aci...

  5. Immunochemical technologies for replacement of rodent bioassays in sensitive detection of toxins in foods

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rapid sensitive assays for biothreat toxins that can be used to detect intentionally contaminated foods are now typically performed via bioassay in live mice. While bioassay provides essential data on bioavailability, animal models are technically, fiscally, and ethically challenging. Through carefu...

  6. Improved high-throughput bioassay for Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As we gain more information through functional genomic studies of Rhyzopertha dominica (F.), we need a high throughput bioassay system to screen potential biopesticides. R. dominica is an internal feeder during immature stages and presents unique challenges with traditional bioassay methods. Our pri...

  7. Development of a High Throughput Translational Bioassay for Plant Biofuel Properties

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Using the well developed microbial system, Clostridium phytofermentans, we have developed a robust bioassay for biomass digestibility and conversion to biofuels. The bioassay can be used to measure the impact of plant genetic diversity on digestibility, and thereby determine the potential effects of...

  8. A Bioassay for Determining Resistance Levels in Tarnished Plant Bug Populations to Neonicotinoid Insecticides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A laboratory bioassay was developed and used to test field populations of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), for resistance development to the neonicitinoid insecticides imidacloprid (Trimax®) and thiamethoxam (Centric®). The bioassay determined LC50 values by feeding...

  9. A LABORATORY BIOASSAY FOR MONITORING RESISTANCE IN TARNISHED PLANT BUG POPULATIONS TO NEONICOTINOID INSECTICIDES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A laboratory bioassay was developed for testing tarnished plant bug populations for resistance development to the neonicotinoid insecticides imidacloprid and thiamethoxam. The bioassay allows for the determination of LC50 values by feeding known doses of the insecticides to adult tarnished plant bu...

  10. Benthic invertebrate bioassays with toxic sediment and pore water

    USGS Publications Warehouse

    Giesy, John P.; Rosiu, Cornell J.; Graney, Robert L.; Henry, Mary G.

    1990-01-01

    The relative sensitivities of bioassays to determine the toxicity of sediments were investigated and three methods of making the sample dilutions required to generate dose-response relationships were compared. The assays studied were: (a) Microtox®, a 15-min assay ofPhotobacterium phosphoreum bioluminescence inhibition by pore water; (b) 48-h Daphnia magnalethality test in pore water; (c) 10-d subchronic assay of lethality to and reduction of weight gain by Chironomus tentans performed in either whole sediment or pore water; (d) 168-h acute lethality assay of Hexagenia limbata in either whole sediment or pore water. The three methods of diluting sediments were: (a) extracting pore water from the toxic location and dilution with pore water from the control station; (b) diluting whole sediment from the toxic location with control whole sediment from a reference location, then extracting pore water; and (c) diluting toxic, whole sediment with whole sediment from a reference location, then using the whole sediment in bioassays. Based on lethality, H. limbata was the most sensitive organism to the toxicity of Detroit River sediment. Lethality of D. magna in pore water was similar to that of H. limbata in whole sediment and can be used to predict effects of whole sediment toxicity to H. limbata. The concentration required to cause a 50% reduction in C. tentans growth (10-d EC50) was approximately that which caused 50% lethality of D. magna (48-h LC50) and was similar to the toxicity that restricts benthic invertebrate colonization of contaminated sediments. While the three dilution techniques gave similar results with some assays, they gave very different results in other assays. The dose-response relationships determined by the three dilution techniques would be expected to vary with sediment, toxicant and bioassay type, and the dose-response relationship derived from each technique needs to be interpreted accordingly.

  11. Evaluation of the mutagenicity and carcinogenicity of motor vehicle emissions in short-term bioassays.

    PubMed Central

    Lewtas, J

    1983-01-01

    Incomplete combustion of fuel in motor vehicles results in the emission of submicron carbonaceous particles which, after cooling and dilution, contain varying quantities of extractable organic constituents. These organics are mutagenic in bacteria. Confirmatory bioassays in mammalian cells provide the capability of detecting chromosomal and DNA damage in addition to gene mutations. In order to evaluate the mutagenicity of these organics in mammalian cells, extractable organics from particle emissions from several diesel and gasoline vehicles were compared in a battery of microbial, mammalian cell and in vivo bioassays. The mammalian cell mutagenicity bioassays were selected to detect gene mutations, DNA damage, and chromosomal effects. Carcinogenesis bioassays conducted included short-term assays for oncogenic transformation and skin tumorigenesis. The results in different assay systems are compared both qualitatively and quantitatively. Good quantitative correlations were observed between several mutagenesis and carcinogenesis bioassays for this series of diesel and gasoline emissions. PMID:6186475

  12. Chronic and Initiation/Promotion Skin Bioassays of Petroleum Refinery Streams.

    PubMed Central

    Skisak, C; Furedi-Machacek, EM; Schmitt, SS; Swanson, MS; Vernot, EH

    1994-01-01

    Nine refinery streams were tested in both chronic and initiation/promotion (I/P) skin bioassays. In the chronic bioassay, groups of 50 C3H/HeJ mice received twice weekly applications of 50 microl of test article for at least 2 years. In the initiation phase of the I/P bioassay, groups of CD-1 mice received an initiating dose of 50 microl of test article for 5 consecutive days, followed by promotion with 50 microl of phorbol-12-myristate-13-acetate (0.01% w/v in acetone) for 25 weeks. In the promotion phase of the I/P bioassay, CD-1 mice were initiated with 50 microl of 7,12-dimethylbenzanthracene (0.1% w/v in acetone) or acetone, followed by promotion with 50 microl of test article twice weekly for 25 weeks. The most volatile of the streams, sweetened naphtha, and the least volatile, vacuum residuum, were noncarcinogenic in both assays. Middle distillates, with a boiling range of 150 degrees-370 degreesC, demonstrated carcinogenic activity in the chronic bioassay and acted as promoters but not initiators in the I/P bioassay. Untreated mineral oil streams displayed initiating activity and were carcinogenic in the chronic bioassay, presumably due to the presence of polycyclic aromatic hydrocarbons of requisite size and structure. A highly solvent-refined mineral oil stream lacked initiating activity. These results indicate that the I/P bioassay, which takes 6 months to complete, may be a good qualitative predictor of the results of a chronic bioassay, at least for petroleum streams. Furthermore, the I/P bioassay can provide insight into possible mechanisms of tumor development. Images p82-a PMID:9719673

  13. Harvester ant bioassay for assessing hazardous chemical waste sites

    SciTech Connect

    Gano, K.A.; Carlile, D.W.; Rogers, L.E.

    1984-12-01

    A technique was developed for using harvester ants, Pogonomyrmex owhyeei, in terrestrial bioassays. Procedures were developed for maintaining stock populations, handling ants, and exposing ants to toxic materials. Relative toxicities were determined by exposing ants to 10 different materials. These materials included three insecticides, Endrin, Aldrin, and Dieldrin; one herbicide, 2,4-D; three oil-like compounds, wood preservative, drilling fluid, and slop oil; and three heavy metals, copper, zinc, and cadmium. Ants were exposed in petri dishes containing soil amended with a particular toxicant. Under these test conditions, ants showed no sensitivity to the metals or 2,4-D. Ants were sensitive to the insecticides and oils in repeated tests, and relative toxicity remained consistent throughout. Aldrin was the most toxic material, followed by Dieldrin, Endrin, wood preservative, drilling fluid, and slop oil. 10 refs., 2 figs., 2 tabs.

  14. Acute bioassays with benthic macroinvertebrates conducted in situ

    SciTech Connect

    Whaley, M.; Garcia, R.; Sy, J. )

    1989-10-01

    Several methods of toxicity testing using macroinvertebrates in controlled laboratory experiments have been reported. Researchers conducted bioassays with natural assemblages of benthic macroinvertebrates exposed to several petroleum refinery effluents. They found that the populations of invertebrates declined after only a few days of exposure. The objective of the study was to determine the acute toxic effects of discharge water from a petrochemical complex on a natural assemblage of benthic macroinvertebrates. The discharge water consisted of refinery wastewater and sanitary wastewater, as well as brine discharge from a power/desalination plant. The benthic macroinvertebrates were transplanted from a healthy reef area to the outfall channel receiving the discharge water. The study began on October 7, 1985, and concluded that same week. Any decrease in specific species would indicate that the discharge was toxic to these species. These species could also serve as indicators of toxic conditions at other locations.

  15. A sediment suspension system for bioassays with small aquatic organisms

    USGS Publications Warehouse

    Schmidt-Dallmier, M. J.; Atchison, G.J.; Steingraeber, M.T.; Knights, B.C.

    1992-01-01

    Exposure of aquatic organisms to suspended sediments can impair growth and survival and increase bioaccumulation of sediment-associated contaminants. However, evaluation of the effects of suspended sediments and their associated contaminants on aquatic organisms has been hampered by the lack of a practical and inexpensive exposure system for conducting bioassays. We present a cost-effective system for assessing the effects of suspended sediments and associated contaminants on small aquatic organisms. A 7-day suspension test was conducted with nominal sediment concentrations ranging from 0.0 To 5.0 g 1-1. The system maintained relatively constant suspended sediment concentrations, as measured by turbidity, and caused minimal mortality to test organisms.

  16. Use of bioassay methods to evaluate incinerator emissions

    SciTech Connect

    Watts, R.R.; DeMarini, D.M.; Linak, W.P.; Lemieux, P.M.; McSorley, J.A.

    1989-01-01

    The organic components in combustion emissions are composed of thousands of chemicals. Analyzing such a complex mixture for the presence of even a few selected chemicals is difficult and provides information on only a fraction of the chemicals present. Reliance on such limited chemical analysis for determining possible health effects may ignore the contribution of many other chemical components of the effluent. Because combustion emissions are complex mixtures, they have been evaluated as such, rather than by studying a few selected chemicals that might be present. The Salmonella (Ames) assay was used to determine the mutagenicity associated with particles from the effluent of municipal-waste combustors, from ambient air collected near a municipal-waste combustor, and from the effluent of a pilot-sized rotary kiln in which polyethylene was combusted. Filter samples were extracted with dichloromethane, and concentrated extracts were solvent exchanged into dimethyl sulfoxide for bioassay.

  17. Toxicity assessment using different bioassays and microbial biosensors.

    PubMed

    Hassan, Sedky H A; Van Ginkel, Steven W; Hussein, Mohamed A M; Abskharon, Romany; Oh, Sang-Eun

    2016-01-01

    Toxicity assessment of water streams, wastewater, and contaminated sediments, is a very important part of environmental pollution monitoring. Evaluation of biological effects using a rapid, sensitive and cost effective method can indicate specific information on ecotoxicity assessment. Recently, different biological assays for toxicity assessment based on higher and lower organisms such as fish, invertebrates, plants and algal cells, and microbial bioassays have been used. This review focuses on microbial biosensors as an analytical device for environmental, food, and biomedical applications. Different techniques which are commonly used in microbial biosensing include amperometry, potentiometry, conductometry, voltammetry, microbial fuel cells, fluorescence, bioluminescence, and colorimetry. Examples of the use of different microbial biosensors in assessing a variety of environments are summarized. PMID:27071051

  18. A novel bioassay using root re-growth in Lemna.

    PubMed

    Park, Areum; Kim, Youn-Jung; Choi, Eun-Mi; Brown, Murray T; Han, Taejun

    2013-09-15

    A new phytotoxicity test method based on root elongation of three Lemna species (Lemna gibba, L. minor, and L. paucicostata) has been developed. Tests with aquatic plants have, typically, favored measurements on fronds (e.g. frond number, area, biomass) rather than on roots, due, in part, to issues associated with handling fragile roots and the time-consuming procedures of selecting roots with identical root lengths. The present method differs in that roots were excised prior to exposure with subsequent measurements on newly developed roots. Results show that there were species-specific difference in sensitivity to the five metals tested (Ag, Cd, Cr, Cu and Hg), with Ag being the most toxic (EC50=5.3-37.6 μgL(-1)) to all three species, and Cr the least toxic for L. gibba and L. minor (1148.3 and 341.8 μgL(-1), respectively) and Cu for L. paucicostata (470.4 μgL(-1)). Direct comparisons were made with measurements of frond area, which were found to be less sensitive. More generally, root re-growth was shown to reflect the toxic responses of all three Lemna species to these five important metals. The root growth bioassay differs from three internationally standardized methods (ISO, OCED and US EPA) in that it is completed in 48 h, the required volume of test solutions is only 3 ml and non-axenic plants are used. Our results show that the Lemna root method is a simple, rapid, cost-effective, sensitive and precise bioassay to assess the toxic risks of metals and has practical application for monitoring municipal and industrial waste waters where metals are common constituents. PMID:23917640

  19. Comprehensive integration of homogeneous bioassays via centrifugo-pneumatic cascading.

    PubMed

    Godino, Neus; Gorkin, Robert; Linares, Ana V; Burger, Robert; Ducrée, Jens

    2013-02-21

    This work for the first time presents the full integration and automation concept for a range of bioassays leveraged by cascading a centrifugo-pneumatic valving scheme to sequentially move several liquids through shared channel segments for multi-step sample preparation into the detection zone. This novel centrifugo-pneumatic liquid handling significantly simplifies system manufacture by obviating the need for complex surface functionalization procedures or hybrid material integration, as it is common in conventional valving methods such as capillary burst valves or sacrificial valves. Based on the centrifugo-pneumatic valving scheme, this work presents a toolkit of operational elements implementing liquid loading/transfer, metering, mixing and sedimentation in a microstructured polymer disc. As a proof of concept for the broad class of homogeneous bioassays, the full integration and automation of a colorimetric nitrate/nitrite test for the detection of clinically relevant nitric oxide (NO) in whole blood is implemented. First, 40 μL of plasma is extracted from a 100 μL sample of human blood, incubated for one hour with the enzymatic mixture (60 μL), and finally reacted with 100 μL of colorimetric (Greiss) reagents. Following just a single loading phase at the beginning of the process, all of these steps are automated through the centrifugo-pneumatic cascade with a high level of flow control and synchronization. Our system shows good correlation with controls up to 50 μM of nitrate, which adequately covers the healthy human range (4 to 45.3 μM). PMID:23250328

  20. Using lone star ticks, Amblyomma americanum (Acari: Ixodidae) in in vitro laboratory bioassays of repellents: dimensions, duration, and variability

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The in vitro bioassay is an important tool in repellent discovery and development, with a variety of bioassays used in recent years. Several factors, such as the dimensions and configuration of test surfaces and duration of tick exposure, can influence the outcome of bioassays. We tested two tick re...

  1. An in vitro rainbow trout cell bioassay for AhR-mediated toxins

    SciTech Connect

    Richter, C.A.; Giesy, J.P.; Denison, M.S.

    1995-12-31

    The toxicity of PCBs, dioxins, and other halogenated aromatic hydrocarbons (HAHS) at environmentally relevant concentrations is in large part mediated through the aromatic hydrocarbon receptor (AhR). Bioassays which measure the activity of genes regulated by the receptor provide an integrative measure of the total AhR-mediated toxicity of a sample. The authors have recently developed and characterized a bioassay using recombinant rainbow trout hepatoma cells containing the firefly luciferase reporter gene under the regulation of the AhR. The cell line is designated Remodulated Lightning Trout (RLT). The RLT bioassay is relevant to fish, and is useful as a rapid screening device, a guide for chemical analysis, and a tool for studies of the AhR mechanism. The responses of the RLT cell line to various PCB congeners are similar to responses of in vivo fish bioassays. The authors now report on the responses of the bioassay to dioxins, dibenzofurans, and other related compounds as compared to in vivo fish bioassays. The authors will also report on the utility of the RLT bioassay in measuring the total TEQ of complex mixtures.

  2. Evaluation and simplification of the assimilable organic carbon nutrient bioassay for bacterial growth in drinking water.

    PubMed

    Kaplan, L A; Bott, T L; Reasoner, D J

    1993-05-01

    A modified assimilable organic carbon (AOC) bioassay is proposed. We evaluated all aspects of the AOC bioassay technique, including inoculum, incubation water, bioassay vessel, and enumeration technique. Other concerns included eliminating the need to prepare organic carbon-free glassware and minimizing the risks of bacterial and organic carbon contamination. Borosilicate vials (40 ml) with Teflon-lined silicone septa are acceptable incubation vessels. Precleaned vials are commercially available, and the inoculum can be injected directly through the septa. Both bioassay organisms, Pseudomonas fluorescens P-17 and Spirillum sp. strain NOX, are available from the American Type Culture Collection and grow well on R2A agar, making this a convenient plating medium. Turbid raw waters need to be filtered prior to an AOC analysis. Glass fiber filters used with either a peristaltic pump or a syringe-type filter holder are recommended for this purpose. A sampling design that emphasizes replication of the highest experimental level, individual batch cultures, is the most efficacious way to reduce the total variance associated with the AOC bioassay. Quality control for the AOC bioassay includes an AOC blank and checks for organic carbon limitation and inhibition of the bioassay organisms. PMID:8517748

  3. Sensitive, Rapid, and Specific Bioassay for the Determination of Antilipogenic Compounds

    PubMed Central

    Ulitzur, S.; Goldberg, I.

    1977-01-01

    A sensitive and rapid bioassay for the determination of the antilipogenic compounds cerulenin and CM-55 is described. The bioassay is based on the inhibitory effect of cerulenin and CM-55 on the in vivo luminescence of an aldehyde-requiring mutant of the marine bacterium Beneckea harveyi. A total quantity as low as 0.1 μg of cerulenin can be determined within 15 min with an error of ±2%. The bioassay, as presented, is specific for compounds that are known to inhibit fatty acid biosynthesis and, as such, it might be used as a general screening method for the detection of antilipogenic compounds. PMID:303076

  4. Toxicity assessment for petroleum-contaminated soil using terrestrial invertebrates and plant bioassays.

    PubMed

    Hentati, Olfa; Lachhab, Radhia; Ayadi, Mariem; Ksibi, Mohamed

    2013-04-01

    The assessment of soil quality after a chemical or oil spill and/or remediation effort may be measured by evaluating the toxicity of soil organisms. To enhance our understanding of the soil quality resulting from laboratory and oil field spill remediation, we assessed toxicity levels by using earthworms and springtails testing and plant growth experiments. Total petroleum hydrocarbons (TPH)-contaminated soil samples were collected from an oilfield in Sfax, Tunisia. Two types of bioassays were performed. The first assessed the toxicity of spiked crude oil (API gravity 32) in Organization for Economic Co-operation and Development artificial soil. The second evaluated the habitat function through the avoidance responses of earthworms and springtails and the ability of Avena sativa to grow in TPH-contaminated soils diluted with farmland soil. The EC50 of petroleum-contaminated soil for earthworms was 644 mg of TPH/kg of soil at 14 days, with 67 % of the earthworms dying after 14 days when the TPH content reached 1,000 mg/kg. The average germination rate, calculated 8 days after sowing, varied between 64 and 74 % in low contaminated soils and less than 50 % in highly contaminated soils. PMID:22773148

  5. Methods to improve routine bioassay monitoring for freshly separated, poorly transported plutonium

    SciTech Connect

    Bihl, D.E.; Lynch, T.P.; Carbaugh, E.H.; Sula, M.J.

    1988-09-01

    Several human cases involving inhalation of plutonium oxide at Hanford have shown clearance half-times from the lung that are much longer than the 500-day half-time recommended for class Y plutonium in Publication 30 of the International Commission on Radiological Protection(ICRP). The more tenaciously retained material is referred to as super class Y plutonium. The ability to detect super class Y plutonium by current routine bioassay measurements is shown to be poor. Pacific Northwest Laboratory staff involved in the Hanford Internal Dosimetry Program investigated four methods to se if improvements in routine monitoring of workers for fresh super class Y plutonium are feasible. The methods were lung counting, urine sampling, fecal sampling, and use of diethylenetriaminepentaacetate (DTPA) to enhance urinary excretion. Use of DTPA was determined to be not feasible. Routine fecal sampling was found to be feasible but not recommended. Recommendations were made to improve the detection level for routine annual urinalysis and routine annual lung counting. 12 refs., 9 figs., 7 tabs.

  6. Analyzing bioassay data using Bayesian methods -- A primer

    SciTech Connect

    Miller, G.; Inkret, W.C.; Schillaci, M.E.

    1997-10-16

    The classical statistics approach used in health physics for the interpretation of measurements is deficient in that it does not allow for the consideration of needle in a haystack effects, where events that are rare in a population are being detected. In fact, this is often the case in health physics measurements, and the false positive fraction is often very large using the prescriptions of classical statistics. Bayesian statistics provides an objective methodology to ensure acceptably small false positive fractions. The authors present the basic methodology and a heuristic discussion. Examples are given using numerically generated and real bioassay data (Tritium). Various analytical models are used to fit the prior probability distribution, in order to test the sensitivity to choice of model. Parametric studies show that the normalized Bayesian decision level k{sub {alpha}}-L{sub c}/{sigma}{sub 0}, where {sigma}{sub 0} is the measurement uncertainty for zero true amount, is usually in the range from 3 to 5 depending on the true positive rate. Four times {sigma}{sub 0} rather than approximately two times {sigma}{sub 0}, as in classical statistics, would often seem a better choice for the decision level.

  7. Target organs in chronic bioassays of 533 chemical carcinogens

    SciTech Connect

    Gold, L.S.; Slone, T.H.; Manley, N.B. ); Bernstein, L. )

    1991-06-01

    A compendium of carcinogenesis bioassay results organized by target organ is presented for 533 chemicals that are carcinogenic in at least one species. This compendium is based primarily on experiments in rats or mice; results in hamsters, nonhuman primates, and dogs are also reported. The compendium can be used to identify chemicals that induce tumors at particular sites, and to determine whether target sites are the same for chemicals positive in more than one species. The Carcinogenic Potency Database (CPDB), which includes results of 3969 experiments, is used in the analysis. The published CPDB includes details on each test, and literature references. Chemical carcinogens are reported for 35 different target organs in rats or mice. More than 80% of the carcinogens in each of these species are positive in at least one of the 8 most frequent target sites; liver, lung, mammary gland, stomach, vascular system, kidney, hematopoietic system, and urinary bladder. An analysis is presented of how well one can predict the carcinogenic response in mice from results in rats, or vice versa. Among chemicals tested in both species, 76% of rat carcinogens are positive in mice, and 71% of mouse carcinogens are positive in rats. Prediction is less accurate to the same target site: 52% of rat carcinogens are positive in the same site in mice, and 48% of mouse carcinogens are positive in the same site in rats. The liver is the most frequent site in common between rats and mice.

  8. Using enzyme bioassays as a rapid screen for metal toxicity

    USGS Publications Warehouse

    Choate, LaDonna M.; Ross, P.E.; Blumenstein, E. P.; Ranville, James F.

    2005-01-01

    Mine tailings piles and abandoned mine soils are often contaminated by a suite of toxic metals, which were released in the mining process. Traditionally, toxicity of such areas has been determined by numerous chemical methods including the Toxicity Characteristic Leachate Procedure (TCLP) and traditional toxicity tests using organisms such as the cladoceran Ceriodaphnia dubia. Such tests can be expensive and time-consuming. Enzymatic bioassays may provide an easier, less costly, and more time-effective toxicity screening procedure for mine tailings and abandoned mine soil leachates. This study evaluated the commercially available MetPLATE™ enzymatic toxicity assay test kit. The MetPLATE™ assay uses a modified strain of Escherichia coli bacteria as the test organism. Toxicity is defined by the activity of β-galactosidase enzyme which is monitored colorometrically with a 96-well spectrophotometer. The study used water samples collected from North Fork Clear Creek, a mining influenced water (MIW) located in Colorado. A great benefit to using the MetPLATE™ assay over the TCLP is that it shows actual toxicity of a sample by taking into account the bioavailability of the toxicants rather than simply measuring the metal concentration present. Benefits of the MetPLATE™ assay over the use of C. dubia include greatly reduced time for the testing process (∼2 hours), a more continuous variable due to a greater number of organisms present in each sample (100,000+), and the elimination of need to maintain a culture of organisms at all times.

  9. Bioassays on Illinois waterway dredged material. Final report

    SciTech Connect

    Moore, D.W.; Gibson, A.B.; Dillon, T.M.

    1992-12-01

    Sediment from the Illinois Waterway navigation channel is hydraulically dredged by the US Army Engineer District, Rock Island, and placed in the nearshore environment via pipeline. Water returning to the river can have a high-suspended solids load approaching fluid mud consistency. There is a concern that this return water may exceed the State of Illinois water quality standards for ammonia and have adverse effects on aquatic life. To address these concerns, composite sediment samples and site water collected from selected sites in the Illinois Waterway were evaluated in toxicity tests. Acute (48-hr) toxicity tests were conducted with two species, Pimephales promelas (the fathead minnow) and Daphnia magna (a freshwater cladoceran). A chronic (21-day) toxicity test was also conducted using Daphnia magna. Animals were exposed separately to different concentrations of filtered and unfiltered elutriates prepared from Acute, Cadmium, Daphnia magna, Pimephales promela, Ammonia, Chronic, Elutriate, Sediment, Bioassay, Cladoceran, Fathead minnow. Illinois Waterway edged material. Total ammonia concentrations were measured in all tests and the un-ionized fraction was calculated by adjusting for temperature and pH. Tests were conducted at the US Army Engineer Waterways Experiment Station, Vicksburg, MS. In addition, as part of an interlaboratory effort, a 48-hr acute toxicity test with Pimephales pomelas fry was conducted concurrently by the Hygienic Laboratory of the University of Iowa, Des Moines, IA.

  10. Detection of Organic Compounds with Whole-Cell Bioluminescent Bioassays

    PubMed Central

    Xu, Tingting; Close, Dan; Smartt, Abby; Ripp, Steven

    2015-01-01

    Natural and manmade organic chemicals are widely deposited across a diverse range of ecosystems including air, surface water, groundwater, wastewater, soil, sediment, and marine environments. Some organic compounds, despite their industrial values, are toxic to living organisms and pose significant health risks to humans and wildlife. Detection and monitoring of these organic pollutants in environmental matrices therefore is of great interest and need for remediation and health risk assessment. Although these detections have traditionally been performed using analytical chemical approaches that offer highly sensitive and specific identification of target compounds, these methods require specialized equipment and trained operators, and fail to describe potential bioavailable effects on living organisms. Alternatively, the integration of bioluminescent systems into whole-cell bioreporters presents a new capacity for organic compound detection. These bioreporters are constructed by incorporating reporter genes into catabolic or signaling pathways that are present within living cells and emit a bioluminescent signal that can be detected upon exposure to target chemicals. Although relatively less specific compared to analytical methods, bioluminescent bioassays are more cost-effective, more rapid, can be scaled to higher throughput, and can be designed to report not only the presence but also the bioavailability of target substances. This chapter reviews available bacterial and eukaryotic whole-cell bioreporters for sensing organic pollutants and their applications in a variety of sample matrices. PMID:25084996

  11. Bioassay-based risk assessment of complex mixtures

    SciTech Connect

    Donnelly, K.C.; Safe, S.H.; Randerath, K.; Randerath, E.

    1994-12-31

    To compare the standard chemical-based risk assessment with in vitro genotoxicity assays, two complex environmental mixtures from a wood preserving site were analyzed in the Salmonella/microsome and E. coli prophage induction assays. Using GC/MS, sample 003 was found to contain relatively low levels of polycyclic aromatic hydrocarbons (PNAs) and elevated levels of polychlorinated dibenzo-p-dioxins (PCDDs), while sample 005 had higher levels of PNAs and relatively low levels of PCDDs. The complex mixtures were sequentially extracted with methylene chloride and methanol for analysis in Salmonella, or extracted with 1:1 hexane: acetone mixture for analysis in the prophage induction assay. At a dose of 1.0 mg/plate in Salmonella strain TA98 with metabolic activation, the methanol extract of sample 003 induced 197 net revertants, while sample 005 induced 436 net revertants. In the prophage induction assay, with activation, the hexane:acetone extract of sample 003 induced a fold increase that was slightly lower than that observed with sample 005. The estimated incremental carcinogenic risk for dermal adsorption and ingestion was 1.5E-3 for sample 003, while for sample 005 the estimated risk was 1.5E-2. Thus, the sample which induced the maximum response in both bioassays also had the highest estimated cancer risk. However, the frequency of PNA-DNA adducts in both skin and liver tissues was appreciably higher with sample 005 than with sample 003.

  12. Target organs in chronic bioassays of 533 chemical carcinogens.

    PubMed Central

    Gold, L S; Slone, T H; Manley, N B; Bernstein, L

    1991-01-01

    A compendium of carcinogenesis bioassay results organized by target organ is presented for 533 chemicals that are carcinogenic in at least one species. This compendium is based primarily on experiments in rats or mice; results in hamsters, nonhuman primates, and dogs are also reported. The compendium can be used to identify chemicals that induce tumors at particular sites, and to determine whether target sites are the same for chemicals positive in more than one species. The Carcinogenic Potency Database (CPDB), which includes results of 3969 experiments, is used in the analysis. The published CPDB includes details on each test, and literature references. Chemical carcinogens are reported for 35 different target organs in rats or mice. More than 80% of the carcinogens in each of these species are positive in at least one of the 8 most frequent target sites: liver, lung, mammary gland, stomach, vascular system, kidney, hematopoietic system, and urinary bladder. An analysis is presented of how well one can predict the carcinogenic response in mice from results in rats, or vice versa. Among chemicals tested in both species, 76% of rat carcinogens are positive in mice, and 71% of mouse carcinogens are positive in rats. Prediction is less accurate to the same target site: 52% of rat carcinogens are positive in the same site in mice, and 48% of mouse carcinogens are positive in the same site in rats. The liver is the most frequent site in common between rats and mice. PMID:1773795

  13. Bioassay-based risk assessment of complex mixtures

    SciTech Connect

    Donnelly, K.C.; Huebner, H.J.

    1996-12-31

    The baseline risk assessment often plays an integral role in various decision-making processes at Superfund sites. The present study reports on risk characterizations prepared for seven complex mixtures using biological and chemical analysis. Three of the samples (A, B, and C) were complex mixtures of polycyclic aromatic hydrocarbons (PAHs) extracted from coal tar; while four samples extracted from munitions-contaminated soil contained primarily nitroaromatic hydrocarbons. The chemical-based risk assessment ranked sample C as least toxic, while the risk associated with samples A and B was approximately equal. The microbial bioassay was in general agreement for the coal tar samples. The weighted activity of the coal tar extracts in Salmonella was 4,960 for sample C, and 162,000 and 206,000 for samples A and B, respectively. The bacterial mutagenicity of 2,4,6-trinitrotoluene contaminated soils exhibited an indirect correlation with chemical-based risk assessment. The aqueous extract of sample 004 induced 1,292 net revertants in Salmonella, while the estimated risk to ingestion and dermal adsorption was 2E-9. The data indicate that the chemical-based risk assessment accurately predicted the genotoxicity of the PAHs, while the accuracy of the risk assessment for munitions contaminated soils was limited due to the presence of metabolites of TNT degradation. The biological tests used in this research provide a valuable compliment to chemical analysis for characterizing the genotoxic risk of complex mixtures.

  14. Colorimetric paper bioassay for the detection of phenolic compounds.

    PubMed

    Alkasir, Ramiz S J; Ornatska, Maryna; Andreescu, Silvana

    2012-11-20

    A new type of paper based bioassay for the colorimetric detection of phenolic compounds including phenol, bisphenol A, catechol and cresols is reported. The sensor is based on a layer-by-layer (LbL) assembly approach formed by alternatively depositing layers of chitosan and alginate polyelectrolytes onto filter paper and physically entrapping the tyrosinase enzyme in between these layers. The sensor response is quantified as a color change resulting from the specific binding of the enzymatically generated quinone to the multilayers of immobilized chitosan on the paper. The color change can be quantified with the naked eye but a digitalized picture can also be used to provide more sensitive comparison to a calibrated color scheme. The sensor was optimized with respect to the number of layers, pH, enzyme, chitosan and alginate amounts. The colorimetric response was concentration dependent, with a detection limit of 0.86 (±0.1) μg/L for each of the phenolic compounds tested. The response time required for the sensor to reach steady-state color varied between 6 and 17 min depending on the phenolic substrate. The sensor showed excellent storage stability at room temperature for several months (92% residual activity after 260 days storage) and demonstrated good functionality in real environmental samples. A procedure to mass-produce the bioactive sensors by inkjet printing the LbL layers of polyelectrolyte and enzyme on paper is demonstrated. PMID:23113670

  15. USING BIOASSAYS TO EVALUATE THE PERFORMANCE OF EDC RISK MANAGEMENT METHODS

    EPA Science Inventory

    In Superfund risk management research, the performance of risk management techniques is typically evaluated by measuring "the concentrations of the chemicals of concern before and after risk management efforts. However, using bioassays and chemical data provides a more robust und...

  16. COMPARATIVE POTENCY OF COMPLEX MIXTURES: USE OF SHORT-TERM GENETIC BIOASSAYS IN CANCER RISK ASSESSMENT

    EPA Science Inventory

    The primary problem regarding the introduction of new energy sources is whether they will alter the mutagenicity, carcinogenicity and potential human cancer risk from combustion emissions. New risk assessment methodologies utilizing data from short-term bioassays, therefore, are ...

  17. IN SITU BIOASSAY CHAMBER FOR ASSESSMENT OF SEDIMENT TOXICITY AND BIOACCUMULATION USING BENTHIC INVERTEBRATES

    EPA Science Inventory

    In this study, we describe the construction of a simple, inexpensive bioassay chamber for testing sediment toxicity (survival and growth) and bioaccumulation under field conditions using the midge Chironomus tentans and the oligochaete Lumbriculus variegatus. The test chamber is ...

  18. DEVELOPMENT OF BIOASSAY PROCEDURES FOR DEFINING POLLUTION OF HARBOR SEDIMENTS. PART I

    EPA Science Inventory

    This research investigates bioassay methods which may be useful in assessing the degree of pollution of harbor sediments. Procedures studied include 96 hr. toxicity tests employing Hexagenia limbata, Daphnia magna and Pontoporeia affinis as biological probes, monitoring cough fre...

  19. CHARACTERIZING THE GENOTOXICITY OF HAZARDOUS INDUSTRIAL WASTES AND EFFLUENTS USING SHORT-TERM BIOASSAYS

    EPA Science Inventory

    This chapter demonstrates that short-term bioassays can reliably and expeditiously measure the genotoxic potential of hazardous industrial wastes and effluents. etrochemical wastes have been studied in detail, especially discharges from chemical manufacturing plants and textile a...

  20. EVALUATION OF THE MUTAGENICITY AND CARCINOGENICITY OF MOTOR VEHICLE EMISSIONS IN SHORT-TERM BIOASSAYS

    EPA Science Inventory

    Incomplete combustion of fuel in motor vehicles results in the emission of submicron carbonaceous particles which, after cooling and dilution, contain varying quantities of extractable organic constituents. These organics are mutagenic in bacteria. Confirmatory bioassays in mamma...

  1. Comparative susceptibility of bemisia tabaci to imidacloprid in field- and laboratory-based bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bemisia tabaci biotype B is a resistance-prone pest of protected and open agriculture. Systemic uptake bioassays used in resistance monitoring programs have provided important information on susceptibility to neonicotinoid insecticides, but have remained decoupled from field performance. Simultaneou...

  2. EVALUATION OF THREE FISH SPECIES AS BIOASSAY ORGANISMS FOR DREDGED MATERIAL TESTING

    EPA Science Inventory

    Three fish species, Cyprinodon variegatus, Fundulus similis, and Menidia menidia, were evaluated to determine which is most suitable as a bioassay organism for solid phase testing of dredged material. Acute toxicity and bioaccumulation of polychlorinated biphenyls (PCBs) were mon...

  3. Harmonia Axyridis Adults Avoid Catnip and Grapefruit-derived Terpenoids in Laboratory Bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We observed the avoidance behavior of the multicolored Asian lady beetle, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), when adults were exposed to volatiles derived from catnip oil and grapefruit seed. In replicated laboratory bioassays, beetles avoided contact with volatiles emanating f...

  4. Evaluation of toxicity of selected insecticides against thrips on cotton in laboratory bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Adult vial technique (AVT) and spray table bioassays were conducted to evaluate toxicity of selected insecticides against immature and adult Western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). In AVT, technical insecticides comprising of organophosphates (d...

  5. A Standardized Lepidopteran Bioassay to Investigate the Bioactivity of Insecticidal Proteins Produced in Transgenic Crops.

    PubMed

    Graser, Gerson; Walters, Frederick S

    2016-01-01

    Insecticidal bioassays are the only reliable method to investigate the biological activity of an insecticidal protein and therefore provide an essential toolkit for the characterization and potency determination of these proteins. Here we present a standardized method for a lepidopteran larval bioassay, which is optimized to specifically estimate activity of insecticidal proteins produced in transgenic plants. The treatment can be either applied to the surface of the artificial diet, or blended into the diet. PMID:26614295

  6. Issues in weighting bioassay data for use in regressions for internal dose assessments

    SciTech Connect

    Strom, D.J.

    1992-11-01

    For use of bioassay data in internal dose assessment, research should be done to clarify the goal desired, the choice of method to achieve the goal, the selection of adjustable parameters, and on the ensemble of information that is available. Understanding of these issues should determine choices of weighting factors for bioassay data used in regression models. This paper provides an assessment of the relative importance of the various factors.

  7. A simple, rapid bioassay for detecting effects of pollutants on bacteria

    SciTech Connect

    Bauer, N.J.; Seidler, R.J.; Knittel, M.D.

    1981-12-01

    A screening bioassay needs to be rapid, and sensitive. The bioassay is described which is accurate, inexpensive, and which utilizes bacteria as the toxicity predictor. The basis of the test involves measuring the kinetics of dissolved oxygen depletion by a mixed microbial population following exposure to a pollutant and allows results to be obtained in as little as 40 min. Pollutants tested were cadmium, copper, nickel, sulfate, diuron, pentachlorophenol, atrazine, tricholoracetic acid, dimethylformamide, and diazinon. (JMT)

  8. Application of root bioassays to detect nutrient deficiencies in fast-growing trees and agroforestry crops

    SciTech Connect

    Harrison, A.F.; Dighton, J.; Jones, H.E.

    1992-12-31

    A new method for the detection of nutrient deficiencies is outlined and recommended as an alternative to conventional soil and foliar analyses. Bioassays are conducted to measure the uptake and supply of the macronutrients. Examples are quoted of the successful use of this technique with Eucalyptus and Sitka spruce. The bioassays have been shown to give equally good results with a range of tree and ground crops.

  9. In situ bioassay using Chironomus riparius: An intermediate between laboratory and field sediment quality assessments

    SciTech Connect

    Guchte, C. van de; Grootelaar, L.; Naber, A.

    1995-12-31

    Benthic macroinvertebrates like chironomid larvae are important indicators for sediment quality. Both in field surveys and laboratory bioassays effect parameters like abundance, survival, growth, larval development and morphological abnormalities of chironomids are recommended biological endpoints to assess the impact of sediment associated contaminants. Now and then results from field surveys on contaminated sites appeared to differ from results in laboratory bioassays on sediment field samples from the same sites. The impact of so-called modifying factors like temperature, oxygen levels and the availability of food could be studied in the laboratory. However, these factors could not fully explain the observed differences. In situ bioassays have been developed to bridge the gap between laboratory and field derived data with respect to the exposure of cultured Chironomus riparius larvae versus field collected Chironomus sp. larvae. Control survival in the in situ bioassays was within acceptable limits (> 80%). Effects observed during the caged exposure of laboratory cultured first instar larvae at contaminated sites were in agreement with the hypothesis that adequate in-field bioassessment reduces uncertainties inherent in the use of standardized laboratory bioassays. Although relative risk ranking of chemicals or contaminated sites can rely upon standard testing protocols, in situ bioassays can give a better insight in exposure-effect relationships under actual field conditions.

  10. An overview of the PubChem BioAssay resource.

    PubMed

    Wang, Yanli; Bolton, Evan; Dracheva, Svetlana; Karapetyan, Karen; Shoemaker, Benjamin A; Suzek, Tugba O; Wang, Jiyao; Xiao, Jewen; Zhang, Jian; Bryant, Stephen H

    2010-01-01

    The PubChem BioAssay database (http://pubchem.ncbi.nlm.nih.gov) is a public repository for biological activities of small molecules and small interfering RNAs (siRNAs) hosted by the US National Institutes of Health (NIH). It archives experimental descriptions of assays and biological test results and makes the information freely accessible to the public. A PubChem BioAssay data entry includes an assay description, a summary and detailed test results. Each assay record is linked to the molecular target, whenever possible, and is cross-referenced to other National Center for Biotechnology Information (NCBI) database records. 'Related BioAssays' are identified by examining the assay target relationship and activity profile of commonly tested compounds. A key goal of PubChem BioAssay is to make the biological activity information easily accessible through the NCBI information retrieval system-Entrez, and various web-based PubChem services. An integrated suite of data analysis tools are available to optimize the utility of the chemical structure and biological activity information within PubChem, enabling researchers to aggregate, compare and analyze biological test results contributed by multiple organizations. In this work, we describe the PubChem BioAssay database, including data model, bioassay deposition and utilities that PubChem provides for searching, downloading and analyzing the biological activity information contained therein. PMID:19933261

  11. A novel bioassay for high-throughput screening microorganisms with N-acyl homoserine lactone degrading activity.

    PubMed

    Liu, Pengfu; Gao, Yang; Huang, Wei; Shao, Zongze; Shi, Jiping; Liu, Ziduo

    2012-05-01

    A novel biosensor strain (Escherichia coli ALM403) that responded to N-acyl homoserine lactone (AHL) was constructed using a luxR-Plux cassette as a regulatory sequence and β-mannanase as a reporter gene. Dinitrosalicylic acid method was used to detect the response of the sensor strain to N-acyl homoserine lactone. By investigating the response to a range of concentrations of N-β-oxooctanoyl-L-homoserine lactone (OOHL), it was demonstrated that the expression of mannanase in E. coli ALM403 could be greatly enhanced by OOHL and resulted in an assayable phenotype. A high-throughput screening approach was developed to isolate AHL-degrading microorganisms, and a marine Halomonas sp. S66-4 showing a marked AHL-degrading ability was successfully isolated. In conclusion, the bioassay system provided a simple and efficient approach to isolate AHL-degrading bacteria. PMID:22528649

  12. Bioassay of thermal protection afforded by candidate flight suit fabrics.

    PubMed

    Knox, F S; Wachtel, T L; McCahan, G R

    1979-10-01

    The United States Army Aeromedical Research Laboratory (USAARL) porcine cutaneous bioassay technique was used to determine what mitigating effect four thermally protective flight suit fabrics would have on fire-induced skin damage. The fabrics were 4.8-ox twill weave Nomex aramide, 4.5-oz stabilized twill weave polybenzimidazole, 4.8-oz plain weave experimental high-temperature polymer (HT4), and 4.8-oz plain weave Nomex aramide (New Weave Nomex or NWN). Each fabric sample was assayed 20 times in each of four configurations: as a single layer in contact with the skin; as a single layer with a 6.35 mm (0.25 in) air gap between fabric and skin; in conjuction with a cotton T-shirt with no air gaps; and, finally, in conjuction with a T-shirt with a 6.35 mm air gap between T-shirt and fabric. Bare skin was used as a control. A JP-4 fueled furnace was used as a thermal source and was adjested to deliver a mean heat flux of 3.07 cal/cm2/s. The duration of exposure was 5 s. Four hundred burn sites were graded using clinical observation and microscopic techniques. Used as single layers, none of the fabrics demonstrated superiority in providing clinically significant protection. When used with a cotton T-shirt, protection was improved. Protection improved progressively for all fabrics and configuration when an air gap was introduced. The experimental high-temperature polymer consistently demonstrated lower heat flux transmission in all configurations, but did not significantly reduce clinical burns. PMID:518445

  13. [Bioassay for enrich-blood bioactivity of Agelicae Sinensis Radix].

    PubMed

    Wang, Xiao-xiao; Zhang, Li-hong; Li, Xi; Wang, Ye; Rong, Zu-yuan; Wei, Hong-ping; Song, Qi-rui; Lv, Guang-hua

    2015-04-01

    Danggui, Agelicae Sinensis Radix, is a widely used Chinese herb to enrich blood, but its quality cannot be effectively assessed by the known chemical markers such as ferulic acid, ligustilide, polysaccharides, etc. A new bioassay was therefore developed to quantify the Enrich-Blood Bioactivity (EBB) for the quality assessment of Danggui raw materials. Danggui sample was first extracted with ethanol and water, respectively. Then the ethanolic extract and water extract were mixed as a test sample to quantify the amount of EBB by mice experiment. The blood deficiency mode in mice was developed by intraperitoneal injecting cyclophospharmide and phenylhdrazine hydrochloride. The quantity of red blood cell was chosen as EBB marker. Cyclosporine A was chosen as a control substance. EBB in analytes was quantified by the amount reaction of parallel line analysis (3, 3') method. The results indicated that the reliability test for quantifying EBB was passed through and the measured value was valid. The analytes showed the significant EBB (P < 0.05). The correlation coefficient was 0.9984 (n=5) between the amount of cyclosporine A (0.035-0.56 g x kg(-1)) and the increased number of red blood cell. The relative standard deviation (RSY) on the amount of EBB was estimated to be 6.15% (n = 6) by six replicated tests, and the confidence limit rate was 26.68% (n = 6). Five Danggui samples, which were collected from different cultivation areas with various morphological characters, showed the variety of EBB in the range of 21.95-44.16 U x g(-1). It is concluded that the developed method is accurate to quantify the EBB of Danggui raw materials, and is therefore suitable to assess its quality. PMID:26281565

  14. Episodic acidification of small streams in the northeastern united states: Fish mortality in field bioassays

    USGS Publications Warehouse

    Van Sickle, J.; Baker, J.P.; Simonin, H.A.; Baldigo, Barry P.; Kretser, W.A.; Sharpe, W.E.

    1996-01-01

    In situ bioassays were performed as part of the Episodic Response Project, to evaluate the effects of episodic stream acidification on mortality of brook trout (Salvelinus fontinalis) and forage fish species. We report the results of 122 bioassays in 13 streams of the three study regions: the Adirondack mountains of New York, the Catskill mountains of New York, and the Northern Appalachian Plateau of Pennsylvania. Bioassays during acidic episodes had significantly higher mortality than did bioassays conducted under nonacidic conditions, but there was little difference in mortality rates in bioassays experiencing acidic episodes and those experiencing acidic conditions throughout the test period. Multiple logistic regression models were used to relate bioassay mortality rates to summary statistics of time-varying stream chemistry (inorganic monomeric aluminum, calcium, pH, and dissolved organic carbon) estimated for the 20-d bioassay periods. The large suite of candidate regressors also included biological, regional, and seasonal factors, as well as several statistics summarizing various features of aluminum exposure duration and magnitude. Regressor variable selection and model assessment were complicated by multicol-linearity and overdispersion. For the target fish species, brook trout, bioassay mortality was most closely related to time-weighted median inorganic aluminum. Median Ca and minimum pH offered additional explanatory power, as did stream-specific aluminum responses. Due to high multicollinearity, the relative importance of different aluminum exposure duration and magnitude variables was difficult to assess, but these variables taken together added no significant explanatory power to models already containing median aluminum. Between 59 and 79% of the variation in brook trout mortality was explained by models employing between one and five regressors. Simpler models were developed for smaller sets of bioassays that tested slimy and mottled sculpin

  15. Benchmarking organic micropollutants in wastewater, recycled water and drinking water with in vitro bioassays.

    PubMed

    Escher, Beate I; Allinson, Mayumi; Altenburger, Rolf; Bain, Peter A; Balaguer, Patrick; Busch, Wibke; Crago, Jordan; Denslow, Nancy D; Dopp, Elke; Hilscherova, Klara; Humpage, Andrew R; Kumar, Anu; Grimaldi, Marina; Jayasinghe, B Sumith; Jarosova, Barbora; Jia, Ai; Makarov, Sergei; Maruya, Keith A; Medvedev, Alex; Mehinto, Alvine C; Mendez, Jamie E; Poulsen, Anita; Prochazka, Erik; Richard, Jessica; Schifferli, Andrea; Schlenk, Daniel; Scholz, Stefan; Shiraishi, Fujio; Snyder, Shane; Su, Guanyong; Tang, Janet Y M; van der Burg, Bart; van der Linden, Sander C; Werner, Inge; Westerheide, Sandy D; Wong, Chris K C; Yang, Min; Yeung, Bonnie H Y; Zhang, Xiaowei; Leusch, Frederic D L

    2014-01-01

    Thousands of organic micropollutants and their transformation products occur in water. Although often present at low concentrations, individual compounds contribute to mixture effects. Cell-based bioassays that target health-relevant biological endpoints may therefore complement chemical analysis for water quality assessment. The objective of this study was to evaluate cell-based bioassays for their suitability to benchmark water quality and to assess efficacy of water treatment processes. The selected bioassays cover relevant steps in the toxicity pathways including induction of xenobiotic metabolism, specific and reactive modes of toxic action, activation of adaptive stress response pathways and system responses. Twenty laboratories applied 103 unique in vitro bioassays to a common set of 10 water samples collected in Australia, including wastewater treatment plant effluent, two types of recycled water (reverse osmosis and ozonation/activated carbon filtration), stormwater, surface water, and drinking water. Sixty-five bioassays (63%) showed positive results in at least one sample, typically in wastewater treatment plant effluent, and only five (5%) were positive in the control (ultrapure water). Each water type had a characteristic bioanalytical profile with particular groups of toxicity pathways either consistently responsive or not responsive across test systems. The most responsive health-relevant endpoints were related to xenobiotic metabolism (pregnane X and aryl hydrocarbon receptors), hormone-mediated modes of action (mainly related to the estrogen, glucocorticoid, and antiandrogen activities), reactive modes of action (genotoxicity) and adaptive stress response pathway (oxidative stress response). This study has demonstrated that selected cell-based bioassays are suitable to benchmark water quality and it is recommended to use a purpose-tailored panel of bioassays for routine monitoring. PMID:24369993

  16. Reading disc-based bioassays with standard computer drives.

    PubMed

    Yu, Hua-Zhong; Li, Yunchao; Ou, Lily M-L

    2013-02-19

    -based bioassays quantitatively. In this Account, we first provide a brief introduction to CD-related materials chemistry and microfluidics research. Then we describe the mild chemistry developed in our laboratory for the preparation of computer-readable biomolecular screening assays: photochemical activation of the polycarbonate (PC) disc surface and immobilization and delivery of probe and target biomolecules. We thoroughly discuss the analysis of the molecular recognition events: researchers can "read" these devices quantitatively with an unmodified optical drive of any personal computer. Finally, and critically, we illustrate our digitized molecular diagnosis approach with three trial systems: DNA hybridization, antibody-antigen binding, and ultrasensitive lead detection with a DNAzyme assay. These examples demonstrate the broad potential of this new analytical/diagnostic tool for medical screening, on-site food/water safety testing, and remote environmental monitoring. PMID:23025412

  17. The usefulness of a sediment bioassay with the gastropod Nassarius reticulatus in tributyltin monitoring programs.

    PubMed

    Laranjeiro, Filipe; Pérez, Sara; Navarro, Patricia; Carrero, José Antonio; Beiras, Ricardo

    2015-11-01

    Despite the use of tributyltin (TBT) had been banned worldwide in 2008 there is still evidence of its deleterious presence in environment. We evaluate the usefulness of a 28days sediment bioassay with Nassarius reticulatus females to monitor TBT pollution, using imposex as endpoint. In addition, butyltins were determined in sediments and tissues, and, whenever posible, imposex was assessed in native N. reticulatus at the same sites where sediments were sampled. In the bioassay, a significant increase in imposex parameters was obtained with three sediments (Vi2, Vi3, and Vi4). No correlation was found between this and TBT concentrations in sediment although good correlations were obtained for TBT in tissues, putting in evidence TBT bioavailability in sediment. A significant decrease in imposex from 2008 to 2013 in native snails was only observed at sites that did not cause any effect in the bioassay. In contrast, imposex levels in 2013 were kept as high as 2008 in one of the sites where a significant imposex increase in the bioassay was observed. The bioassay proves thus to be a practical and ecological relevant tool, as: (i) it can be conducted in sites with no native populations of snails, (ii) it provides early identification of polluted sites, anticipating future imposex levels or early identification of recovering, and (iii) it yields information on the bioavailable fraction of the TBT in the sediment. Therefore, this tool can be of extreme usefulness under the scope of recent European legislative frameworks. PMID:26318117

  18. Rainbow trout cell bioassay-derived relative potencies for halogenated aromatic hydrocarbons: Comparison and sensitivity analysis

    SciTech Connect

    Villeneuve, D.L.; Blankenship, A.L.; Giesy, J.P.; Richter, C.A.

    1999-05-01

    Rainbow trout hepatoma cells, stably transfected with a luciferase reporter gene under control of dioxin-responsive elements (RLT 2.0 cells) were used to derive relative potencies (RPs) for a variety of halogenated aromatic hydrocarbons (HAHs) that are structurally similar to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). This in vitro bioassay utilizes 96-well microplates, which provide high sample throughput and assay efficiency without affecting sensitivity. The RLT 2.0-derived potencies for dioxin and furan congeners, relative to 2,3,7,8-TCDD, ranged from 0.917 for 1,2,3,4,7,8-hexachlorodibenzofuran to 0.208 or 1,2,3,7,8-pentachlorodibenzofuran. All mono- and di-ortho polychlorinated biphenyls (PCBs) tested had RPs that were orders of magnitude less than TCDD, but point estimates could not be determined. The RLT 2.0-derived RPs were found to be comparable to both other rainbow trout-specific RPs and RPs based on mammalian bioassays. Sensitivity analysis suggested that the range of uncertainty associated with TCDD equivalent (TEQ) estimates based on RLT 2.0-derived RPs is approximately 10-fold. Within this degree of uncertainty and the context of this study, the RLT 2.0 bioassay showed no definitive biases or inaccuracies relative to similar mammalian- or fish-specific in vitro bioassays. Thus, the RLT 2.0 bioassay appears to be a useful tool for evaluating dioxin-like potency of HAHs to fish.

  19. Selecting a sensitive battery of bioassays to detect toxic effects of metals in effluents.

    PubMed

    de Paiva Magalhães, Danielly; da Costa Marques, Mônica Regina; Fernandes Baptista, Darcilio; Forsin Buss, Daniel

    2014-09-01

    The use of bioassay batteries is necessary to evaluate toxic effects at various biological levels. The selection of bioassays without prior testing and determination of the most sensitive/suitable groups for each impact may allow the discharge of effluents that pose a threat to the environment. The present study tested and selected a battery of sensitive ecotoxicological bioassays for detecting toxic effects of metals. The sensitivities of six organisms were evaluated (algae Pseudokirchneriella subcapitata and Chlorella vulgaris, Cladocera Daphnia similis and Ceriodaphnia dubia, and fish Poecilia reticulata and Danio rerio) after exposure to 10 individual metal species deemed toxic to the aquatic environment (Ag(+), Cd(2+), Cu(+), Cu(2+), Cr(3+), Cr(6+), Pb(2+), Ni(2+), Zn(2+), and Hg(2+)) and to real (steel-mill) and laboratory simulated effluents. In the bioassays, fish were the least sensitive; D. rerio showed no sensitivity to any of the effluents tested. P. subcapitata was a good bioindicator of Cr(3+) toxicity, and D. similis was the most sensitive organism to Hg(2+); but the toxic effect of effluents with higher levels of Hg(2+) was better detected by C. dubia. The most sensitive battery of bioassays to detect low concentrations of dissolved metals in effluents was the 72-h chronic test with C. vulgaris and the 48-h acute test with C. dubia. PMID:25199585

  20. Characterization of chemical waste site contamination and its extent using bioassays

    SciTech Connect

    Thomas, J.M.; Callahan, C.A.; Cline, J.F.; Greene, J.C.; McShane, M.C.; Miller, W.E.; Peterson, S.A.; Simpson, J.C.; Skalski, J.R.

    1984-12-01

    Bioassays were used in a three-phase research project to assess the comparative sensitivity of test organisms to known chemicals, determine if the chemical components in field soil and water samples containing unknown contaminants could be inferred from our laboratory studies using known chemicals, and to investigate kriging (a relatively new statistical mapping technique) and bioassays as methods to define the areal extent of chemical contamination. The algal assay generally was most sensitive to samples of pure chemicals, soil elutriates and water from eight sites with known chemical contamination. Bioassays of nine samples of unknown chemical composition from the Rocky Mountain Arsenal (RMA) site showed that a lettuce seed soil contact phytoassay was most sensitive. In general, our bioassays can be used to broadly identify toxic components of contaminated soil. Nearly pure compounds of insecticides and herbicides were less toxic in the sensitive bioassays than were the counterpart commercial formulations. This finding indicates that chemical analysis alone may fail to correctly rate the severity of environmental toxicity. Finally, we used the lettuce seed phytoassay and kriging techniques in a field study at RMA to demonstrate the feasibility of mapping contamination to aid in cleanup decisions. 25 references, 9 figures, 9 tables.

  1. Susceptibility of Bagrada hilaris (Hemiptera: Pentatomidae) to Insecticides in Laboratory and Greenhouse Bioassays.

    PubMed

    Palumbo, John C; Prabhaker, Nilima; Reed, Darcy A; Perring, Thomas M; Castle, Steven J; Huang, Ta-I

    2015-04-01

    Field-collected nymphs and adults of Bagrada hilaris (Burmeister) (Hemiptera: Penatatomidae) from three locations were evaluated for susceptibility to insecticides representing 10 classes of insecticide chemistry. Although relative susceptibilities differed between leaf-spray and leaf-dip Petri dish bioassays, consistently low LC50 values were determined for chlorpyrifos, bifenthrin, and lambda-cyhalothrin. Fenpropathrin and methomyl had intermediate values. Susceptibility to dinotefuran varied depending on the bioassay, possibly owing to leaf substrates used in the two bioassays. In soil systemic bioassays, the LC50 value of dinotefuran was significantly greater than that of two other neonicotinoids, imidacloprid and thiamethoxam, and the anthranilic diamide, cyantraniliprole. Mortality and feeding damage of B. hilaris and plant growth on insecticide-treated plants in greenhouse trials were consistent with the laboratory bioassays; the best results were seen with bifenthrin, methomyl, and chlorpyrifos. Mortality to the neonicotinoids was not evident; however, feeding damage and plant growth responses on dinotefuran-treated plants damage were similar to the noninfested control. This highlights the apparent antifeedant properties of dinotefuran that may have prevented adults from injuring broccoli plants after exposure to foliar spray residues. Data presented serve as baseline susceptibilities that can be used to monitor for resistance development in field populations of B. hilaris. PMID:26470178

  2. Studies on the bioassayable growth hormone-like activity of plasma

    NASA Technical Reports Server (NTRS)

    Ellis, S.; Vodian, M. A.; Grindeland, R. E.

    1978-01-01

    Evidence supporting the existence of bioassayable growth hormone-like activity in blood plasma distinct from the growth hormone measurable by radioimmunoassay and from somatomedin is presented. Tibial assays of the growth-hormone-like activity of injected, concentrated normal human and rat plasma in hypophysectomized rats reveal 200- and 50-fold activity excesses, respectively, with respect to the amount of growth hormone detected by radioimmunoassay. The origin of this bioassayable plasma hormone has been localized to the region of the pituitary, the origin of growth hormone, a distribution not followed by somatomedin C. Purification of the bioassayable agent indicates that is has a molecular weight of between 60,000 and 80,000, in contrast to that of growth hormone (20,000), and that the bioassayable activity is distinct from that of somatomedin C. Growth hormone-like activity detected in Cohn fraction IV as well as plasma activity, are found to be collectable on Dowex 50 resin, in contrast to somatomedin C and nonsuppressible insulin-like activity. The formation of bioassayable growth hormone-activity agents from radioimmunoassayable growth hormone and directly in the pituitary is suggested.

  3. Bioaccumulation of organic chemicals in contaminated soils: evaluation of bioassays with earthworms.

    PubMed

    Jager, Tjalling; van der Wal, Leon; Fleuren, Roel H L J; Barendregt, Arjan; Hermens, Joop L M

    2005-01-01

    Earthworms live in close contact with the soil and can thus be considered representative for the bioavailability of chemicals at contaminated sites. Bioavailability can either be assessed by analyzing earthworms from contaminated locations or by exposing laboratory-reared specimens to soil samples from the field (bioassays). In this study, we investigate the relevance of bioassays by using an extended experimental design (to identify signs of depletion of the bioavailable phase by the earthworms) and by using two species of earthworm (the standard test species Eisenia andrei and the field-relevant Aporrectodea caliginosa). Furthermore, bioassay results are compared to body residues of worms collected from the field site: a heavily polluted polder, amended with dredge spoil. We focused on telodrin, dieldrin, hexachlorobenzene, and eight PCBs. With our bioassay design, it was shown that depletion was unlikely, although more subtle effects could have occurred (e.g., changes in sorption during the experiments). E. andrei is a good choice for bioassays because its body residues correlate well to those in A. caliginosa, as well as to those in the field-collected worms. Nevertheless, E. andrei accumulated slightly more than the other species and appeared to be more sensitive to the conditions in soil from one of our sites. PMID:15667108

  4. Bioassay of quinoline, 5-fluoroquinoline, carbazole, 9-methylcarbazole and 9-ethylcarbazole in newborn mice.

    PubMed

    Weyand, E H; Defauw, J; McQueen, C A; Meschter, C L; Meegalla, S K; LaVoie, E J

    1993-10-01

    Quinoline and carbazole are among the more prevalent aza-arenes present as components of environmental pollutants. Both of these aza-arenes are hepatocarcinogenic to mice when administered in the diet. The hepatocarcinogenic potential of quinoline is consistent with its mutagenic activity in Salmonella typhimurium TA100 and potential to induce unscheduled DNA synthesis (UDS) in rat hepatocytes. Structure-activity studies with fluorinated quinolines indicate that the presence of a fluorine atom at the 5-position of quinoline may inhibit detoxification and result in enhanced genotoxic potency. Quinoline and 5-fluoroquinoline were assayed in newborn CD-1 mice at a total dose of 1.75 mumol to establish their relative tumorigenic activity. Liver tumours developed in 60 and 90% of the male newborn mice treated with quinoline and 5-fluoroquinoline, respectively. The majority of liver tumours observed among the quinoline-treated mice were classified as adenomas. In contrast, liver carcinomas developed in most of the male mice treated with 5-fluoroquinoline. Unlike the well established genotoxic potential of quinoline, there is limited evidence for carbazole having either genotoxic or carcinogenic activity. Whereas carbazole is not mutagenic towards several strains of S. typhimurium, both 9-methylcarbazole and 9-ethylcarbazole are active as mutagens in S. typhimurium TA100. Carbazole, 9-methylcarbazole and 9-ethylcarbazole were assayed in primary rat hepatocytes to assess their relative potential to induce UDS in rat hepatocytes; only 9-ethylcarbazole did so. These carbazole derivatives were also assayed in newborn CD-1 mice at a total dose of 1.75 mumol. Neither carbazole nor either of these 9-alkylcarbazoles produced a significant tumorigenic response in this bioassay system. PMID:8225128

  5. Getting ready for the manned mission to Mars: bioassays for space research

    NASA Astrophysics Data System (ADS)

    Baumstark-Khan, Christa; Hellweg, Christine E.; Arenz, Andrea; Meier, Matthias M.; Horneck, Gerda

    2004-06-01

    Harmful environmental factors - namely ionizing radiation - will continue to influence future manned space missions. The Cellular Biodiagnostic group at the German Aerospace Center (DLR) develops cellular monitoring systems, which include bacterial and mammalian cell systems capable of recognizing DNA damage as a consequence of the presence of genotoxic conditions. Such bioassay or biosensor systems will complement the physical detector systems used in space, insofar as they yield intrinsically biologically weighted measures of cellular responses. Furthermore, synergistic mutagenic and cancerogenic impacts of the radiation environment together with other potentially genotoxic constituents of the space habitat can be quantified using such systems, whose signals are especially relevant for the molecular damage to the DNA or the chromosomes. The experiment Cellular Responses to Radiation in Space (CERASP) has been selected by NASA to be performed on the International Space Station. It will supply basic information on the cellular response to radiation applied in microgravity. One of the biological end-points under investigation will be survival reflected by radiation-dependent reduction of constitutive expression of the enhanced variant of green fluorescent protein (EGFP), originally isolated from the bioluminescent jellyfish Aequorea victoria. A second end-point will be gene activation by space flight conditions in mammalian cells, based on fluorescent promoter reporter systems using the destabilized EGFP variant (d2EGFP). The promoter element to be investigated will reflect the activity of the NF-kappaB stress response pathway as an anti-apoptotic radiation response. DNA damage will be measured by fluorescent analysis of DNA unwinding (FADU). The systems have worked properly for terrestrial applications during the first experiments. Experiments using accelerated particles produced at the French heavy ion accelerator GANIL have given insights into cellular mechanisms

  6. Facile synthesis of methotrexate intercalated layered double hydroxides: particle control, structure and bioassay explore.

    PubMed

    Tian, De-Ying; Liu, Zhen-Lei; Li, Shu-Ping; Li, Xiao-Dong

    2014-12-01

    To study the influence of particle size on drug efficacy and other properties, a series of methotrexate intercalated layered double hydroxides (MTX/LDHs) were synthesized through the traditional coprecipitation method, using a mixture of water and polyethylene glycol (PEG-400) as the solvent. To adjust the particle size of MTX/LDHs, the dropping way, the volume ratio of water to PEG-400 and different hydrothermal treatment time changed accordingly, and the results indicate that the particle size can be controlled between 90 and 140 nm. Elemental C/H/N and inductive coupled plasma (ICP) analysis indicated that different synthesis conditions almost have no effect on the compositions of the nanohybrids. X-ray diffraction (XRD) patterns manifested the successful intercalation of MTX anions into the LDH interlayers, and it's also found out that different volume ratios of water to PEG-400 and variable dropping way can affect the crystallinity of the final samples, i.e., the volume ratio of 3:1 and pH decreasing are proved to be optimum conditions. Furthermore, both antiparallel monolayer and bilayers adopting different orientations are suggested for four samples from XRD results. Fourier transform infrared spectroscopy (FTIR) investigations proved the coexistence of CO3(2-) and MTX anions in the interlayer of the nanohybrids. MTX/LDH particles exhibited hexagonal platelet morphology with round corner and different dropping ways can affect the morphology greatly. Moreover, a DSC study indicated that longer time treatment can weaken the bond between the MTX anions and LDH layers. The kinetic release profiles told us that larger MTX/LDH particles have enhanced the ability of LDH layers to protect interlayer molecules. At last, the bioassay study indicated that the nanohybrids with larger diameters have higher tumor suppression efficiency. PMID:25491832

  7. Thioglucose-stabilized gold nanoparticles as a novel platform for colorimetric bioassay based on nanoparticle aggregation.

    PubMed

    Watanabe, Shigeru; Yoshida, Kazuma; Shinkawa, Keitarou; Kumagawa, Daisuke; Seguchi, Hideki

    2010-12-01

    Gold nanoparticles stabilized with thioglucose (TGlu-AuNPs), which have carboxyl groups on the particle surface as anchoring sites for covalent immobilization of biomolecules, were prepared by the chemical reduction of HAuCl4 using 1-thio-β-D-glucose as a reducing and stabilizing agent, and their application to colorimetric bioassay was demonstrated using the carbohydrate-lectin system. p-Aminophenyl α-D-mannose (Man-NH2) was covalently attached by a conventional method to the activated carboxyl groups on the TGlu-AuNPs. On addition of Con A to the Man-AuNPs, multiple binding events occurred between Con A and the mannoses immobilized on the particle surface. This Con A-induced aggregation resulted in a significant red shift in local surface plasmon resonance. The binding isotherm showed a sigmoidal curve, indicating cooperativity in the binding of Con A and the Man-AuNPs. In addition, Hill plots showed two nonequivalent binding modes, with the Kd values for high- and low-affinity binding of 11.3 and 66.5 pM, respectively, which was significantly lower than that for methyl-α-D-mannose binding to Con A. The enhanced binding affinity between Man-AuNPs and Con A involves the cluster effect of the carbohydrate groups on the AuNPs. A linear correlation curve was obtained in the range 10-100 nM (R2=0.983). The limit of detection (LOD) for Con A was 9.0 nM in aqueous buffer, which is comparable to that of other conventional methods such as ELISA. PMID:20801619

  8. A medium-term rat liver bioassay for rapid in vivo detection of carcinogenic potential of chemicals.

    PubMed

    Ito, Nobuyuki; Tamano, Seiko; Shirai, Tomoyuki

    2003-01-01

    A reliable medium-term bioassay system for rapid detection of carcinogenic potential of chemicals in the human environment has been developed. The 8-week-protocol consists of 2 stages; male F344 rats are given a single intraperitoneal injection of diethylnitrosamine (200 mg/kg) for initiation of liver carcinogenesis, followed by a 6-week test chemical treatment starting 2 weeks thereafter. Test chemicals are usually given in the diet or the drinking water and in the 2nd week of test chemical treatment, all rats are subjected to two-thirds partial hepatectomy in order to induce regenerative cell replication. The end-point marker is the glutathione S-transferase placental form (GST-P)-positive hepatic focus, the numbers and sizes of which are analyzed using an image-analyzer and expressed as values per unit liver section (1 cm2). When the yield of GST-P-positive foci is significantly enhanced (P<0.05) over the control value, a chemical is judged to possess carcinogenic or promotion potential for the liver. Among 313 chemicals already tested in this system in our laboratory, 30/31 (97%) mutagenic hepatocarcinogens and 29/33 (88%) non-mutagenic hepatocarcinogens gave positive results. Ten out of 43 (23%) agents known to be carcinogenic in organs other than the liver were also positive. It is particularly important that only one of 48 non-carcinogens gave a very weak positive result, so that the system has a very low false-positivity rate. It is now well documented that the assay system is highly effective for detecting hepatocarcinogens, bridging the gap between traditional long-term carcinogenicity tests and short-term screening assays. At the Fourth International Conference on Harmonization, our medium-term liver bioassay based on an initiation and promotion protocol was recommended in the guidelines as an acceptable alternative to the long-term rodent carcinogenicity test. PMID:12708466

  9. Strategies for Transferring Mixtures of Organic Contaminants from Aquatic Environments into Bioassays.

    PubMed

    Jahnke, Annika; Mayer, Philipp; Schäfer, Sabine; Witt, Gesine; Haase, Nora; Escher, Beate I

    2016-06-01

    Mixtures of organic contaminants are ubiquitous in the environment. Depending on their persistence and physicochemical properties, individual chemicals that make up the mixture partition and distribute within the environment and might then jointly elicit toxicological effects. For the assessment and monitoring of such mixtures, a variety of cell-based in vitro and low-complexity in vivo bioassays based on algae, daphnids or fish embryos are available. A very important and sometimes unrecognized challenge is how to combine sampling, extraction and dosing to transfer the mixtures from the environment into bioassays, while conserving (or re-establishing) their chemical composition at adjustable levels for concentration-effect assessment. This article outlines various strategies for quantifiable transfer from environmental samples including water, sediment, and biota into bioassays using total extraction or polymer-based passive sampling combined with either solvent spiking or passive dosing. PMID:26804122

  10. Bioassay, isolation and studies on the mechanism of action of neurite extension factor

    NASA Technical Reports Server (NTRS)

    Kligman, D.

    1984-01-01

    The identification and purification of molecules active in promoting neurite outgrowth requires a sensitive reproducible bioassay. A quantitative bioassay was utilized to purify a neurite extension factor (NEF) based on counting the number of phase bright neurons with processes at least equal to one cell body diameter after 20 hrs. in culture is defined, serum free medium. Using a combination of heat treatment DEAE cellulose chromatography and gel filtration, an acidic protein of M sub r = 75,000 was highly purified. Upon reduction, it yields subunits of M sub r = 37,000. Purified fractions are active half maximally at 100 ng/ml in inducing neurite outgrowth in this bioassay. Currently, monoclonal antibodies to NEF are being produced. Female Balb C mice were immunized with the antigen and fusions with mouse myeloma cells will be performed to yield hybridoma cells.

  11. Comparison of liquid chromatographic and bioassay procedures for determining depletion of intramuscularly injected tylosin.

    PubMed

    Moats, W A; Harris, E W; Steele, N C

    1985-01-01

    Crossbred pigs weighing 80-110 kg were injected intramuscularly in the ham with 8.8 mg/kg tylosin. Animals were slaughtered in groups of 3 at intervals of 4 h, and 1, 2, 4, and 8 days after injection, and samples of blood, injected muscle, uninjected muscle, liver, and kidney were analyzed by liquid chromatography (LC) and by bioassay using Sarcina lutea as the test organism. The LC method was far more sensitive with a detection limit of less than 0.1 ppm, while the detection limit by bioassay was about 0.5 ppm in tissue. Results by bioassay and LC sometimes differed considerably for tissue samples. Residues in all tissues were below the tolerance limit of 0.2 ppm at 24 h, except in the injected muscle in one animal. Residues were not detected in any tissue of any animal at 48 h after treatment. PMID:4019360

  12. Bioassay for estimating the biogenic methane-generating potential of coal samples

    USGS Publications Warehouse

    Jones, E.J.P.; Voytek, M.A.; Warwick, P.D.; Corum, M.D.; Cohn, A.; Bunnell, J.E.; Clark, A.C.; Orem, W.H.

    2008-01-01

    Generation of secondary biogenic methane in coal beds is likely controlled by a combination of factors such as the bioavailability of coal carbon, the presence of a microbial community to convert coal carbon to methane, and an environment supporting microbial growth and methanogenesis. A set of treatments and controls was developed to bioassay the bioavailability of coal for conversion to methane under defined laboratory conditions. Treatments included adding a well-characterized consortium of bacteria and methanogens (enriched from modern wetland sediments) and providing conditions to support endemic microbial activity. The contribution of desorbed methane in the bioassays was determined in treatments with bromoethane sulfonic acid, an inhibitor of microbial methanogenesis. The bioassay compared 16 subbituminous coal samples collected from beds in Texas (TX), Wyoming (WY), and Alaska (AK), and two bituminous coal samples from Pennsylvania (PA). New biogenic methane was observed in several samples of subbituminous coal with the microbial consortium added, but endemic activity was less commonly observed. The highest methane generation [80????mol methane/g coal (56??scf/ton or 1.75??cm3/g)] was from a south TX coal sample that was collected from a non-gas-producing well. Subbituminous coals from the Powder River Basin, WY and North Slope Borough, AK contained more sorbed (original) methane than the TX coal sample and generated 0-23????mol/g (up to 16??scf/ton or 0.5??cm3/g) new biogenic methane in the bioassay. Standard indicators of thermal maturity such as burial depth, nitrogen content, and calorific value did not explain differences in biogenic methane among subbituminous coal samples. No original methane was observed in two bituminous samples from PA, nor was any new methane generated in bioassays of these samples. The bioassay offers a new tool for assessing the potential of coal for biogenic methane generation, and provides a platform for studying the

  13. In vitro bioassays to evaluate complex chemical mixtures in recycled water

    PubMed Central

    Jia, Ai; Escher, Beate I.; Leusch, Frederic D.L.; Tang, Janet Y.M.; Prochazka, Erik; Dong, Bingfeng; Snyder, Erin M.; Snyder, Shane A.

    2016-01-01

    With burgeoning population and diminishing availability of freshwater resources, the world continues to expand the use of alternative water resources for drinking, and the quality of these sources has been a great concern for the public as well as public health professionals. In vitro bioassays are increasingly being used to enable rapid, relatively inexpensive toxicity screening that can be used in conjunction with analytical chemistry data to evaluate water quality and the effectiveness of water treatment. In this study, a comprehensive bioassay battery consisting of 36 bioassays covering 18 biological endpoints was applied to screen the bioactivity of waters of varying qualities with parallel treatments. Samples include wastewater effluent, ultraviolet light (UV) and/or ozone advanced oxidation processed (AOP) recycled water, and infiltrated recycled groundwater. Based on assay sensitivity and detection frequency in the samples, several endpoints were highlighted in the battery, including assays for genotoxicity, mutagenicity, estrogenic activity, glucocorticoid activity, aryl hydrocarbon receptor activity, oxidative stress response, and cytotoxicity. Attenuation of bioactivity was found to be dependent on the treatment process and bioassay endpoint. For instance, ozone technology significantly removed oxidative stress activity, while UV based technologies were most efficient for the attenuation of glucocorticoid activity. Chlorination partially attenuated genotoxicity and greatly decreased herbicidal activity, while groundwater infiltration efficiently attenuated most of the evaluated bioactivity with the exception of genotoxicity. In some cases, bioactivity (e.g., mutagenicity, genotoxicity, and arylhydrocarbon receptor) increased following water treatment, indicating that transformation products of water treatment may be a concern. Furthermore, several types of bioassays with the same endpoint were compared in this study, which could help guide the selection

  14. Genotoxicity of soil from farmland irrigated with wastewater using three plant bioassays.

    PubMed

    Cabrera, G L; Rodriguez, D M

    1999-05-19

    Three well known plant bioassays, the Allium root chromosome aberration (AL-RAA) assay, the Tradescantia micronucleus (Trad-MCN) assay, and the Tradescantia stamen hair (Trad-SHM) mutation assay were validated in 1991 by the International Programme on Chemical Safety (IPCS) under the auspices of the World Health Organization, and the United Nations Environment Programme (UNEP). These plant bioassays have proven to be efficient tests for chemical screening and especially for in situ monitoring for genotoxicity of environmental pollutants. As a result of this validation study, standard protocols of these three plant bioassays were used by some of the 11 participating countries in the IPCS to carry on genotoxicity tests on air, water and soil as a follow up activity. In the city of Queretaro, Mexico, wastewater coming from both industrial and domestic sources and without any treatment is used to irrigate the farm crops, polluting the soil. Potentially the pollutants could reach the food chain. For the above reason, soil irrigated with wastewater was sampled and monitored for the presence of genotoxic agents using the above three bioassays. Extracts from soil samples were made using distilled water and organic solvents by shaking the sample for about 12 h under a relatively low temperature (15-20 degrees C). Plant cuttings of Tradescantia or the roots of Allium were treated by submerging them in the extracts. Three replicates of each sample were analyzed in each of the three bioassays. Extracts using DMSO, ethanol and distilled water tested positive in the three bioassays and there were no differences for the genotoxicity of the extracts with the different solvents. PMID:10350600

  15. In vitro bioassays to evaluate complex chemical mixtures in recycled water.

    PubMed

    Jia, Ai; Escher, Beate I; Leusch, Frederic D L; Tang, Janet Y M; Prochazka, Erik; Dong, Bingfeng; Snyder, Erin M; Snyder, Shane A

    2015-09-01

    With burgeoning population and diminishing availability of freshwater resources, the world continues to expand the use of alternative water resources for drinking, and the quality of these sources has been a great concern for the public as well as public health professionals. In vitro bioassays are increasingly being used to enable rapid, relatively inexpensive toxicity screening that can be used in conjunction with analytical chemistry data to evaluate water quality and the effectiveness of water treatment. In this study, a comprehensive bioassay battery consisting of 36 bioassays covering 18 biological endpoints was applied to screen the bioactivity of waters of varying qualities with parallel treatments. Samples include wastewater effluent, ultraviolet light (UV) and/or ozone advanced oxidation processed (AOP) recycled water, and infiltrated recycled groundwater. Based on assay sensitivity and detection frequency in the samples, several endpoints were highlighted in the battery, including assays for genotoxicity, mutagenicity, estrogenic activity, glucocorticoid activity, arylhydrocarbon receptor activity, oxidative stress response, and cytotoxicity. Attenuation of bioactivity was found to be dependent on the treatment process and bioassay endpoint. For instance, ozone technology significantly removed oxidative stress activity, while UV based technologies were most efficient for the attenuation of glucocorticoid activity. Chlorination partially attenuated genotoxicity and greatly decreased herbicidal activity, while groundwater infiltration efficiently attenuated most of the evaluated bioactivity with the exception of genotoxicity. In some cases, bioactivity (e.g., mutagenicity, genotoxicity, and arylhydrocarbon receptor) increased following water treatment, indicating that transformation products of water treatment may be a concern. Furthermore, several types of bioassays with the same endpoint were compared in this study, which could help guide the selection

  16. False-Positive Serum Botulism Bioassay in Miller-Fisher Syndrome.

    PubMed

    Zeylikman, Yuriy; Shah, Vishal; Shah, Umang; Mirsen, Thomas R; Campellone, Joseph V

    2015-09-01

    We describe a patient with acute progressive weakness and areflexia. Both botulism and Miller-Fisher variant of Guillain-Barré syndrome were initial diagnostic considerations, and she was treated with intravenous immunoglobulin and botulinum antitoxin. A mouse bioassay was positive for botulinum toxin A, although her clinical course, electrodiagnostic studies, and cerebrospinal fluid findings supported Miller-Fisher syndrome. This patient's atypical features offer points of discussion regarding the evaluation of patients with acute neuromuscular weakness and emphasize the limitations of the botulism bioassay. PMID:26301377

  17. A field bioassay to evaluate potential spatial repellents against natural mosquito populations.

    PubMed

    Chauhan, K R; Aldrich, J R; McCardle, P W; White, G B; Webb, R E

    2012-12-01

    A field bioassay evaluating candidate chemicals as aerial repellents was developed and evaluated against natural mosquito populations in Beltsville, MD. The bioassay consisted of an attractive source surrounded by a grid of 16 septa containing a volatile candidate aerial repellent, compared with an attractive source without such a grid. The attractive source was a Centers for Disease Control and Prevention light trap supplemented with carbon dioxide. Significant sources of variation included weather, position, and the differential response of mosquito species. Despite these sources of variation, significant repellent responses were obtained for catnip oil, E,Z-dihydronepetalactone, and DEET. PMID:23393752

  18. A Brine Shrimp Bioassay for Measuring Toxicity and Remediation of Chemicals

    NASA Astrophysics Data System (ADS)

    Lieberman, Marya

    1999-12-01

    A bioassay using Artemia franciscana (brine shrimp) was adapted to measure the toxicity of household chemicals. One project is described in which students collect dose-response curves for seven commercial flea-killing products. Next, groups of students researched the insecticidal ingredients of the flea products. On the basis of the structures of the active ingredients, they chose remediation methods to make the flea product less toxic to brine shrimp; procedures included copper-catalyzed hydrolysis, adsorption onto activated charcoal, bleach treatment, and photodegradation. No special equipment or supplies are necessary for the bioassay other than the brine shrimp eggs, which can be obtained at any aquarium store.

  19. Review of Bioassays for Monitoring Fate and Transport ofEstrogenic Endocrine Disrupting Compounds in Water

    SciTech Connect

    CGCampbell@lbl.gov

    2004-01-30

    Endocrine disrupting compounds (EDCs) are recognizedcontaminants threatening water quality. Despite efforts in sourceidentification, few strategies exist for characterization or treatment ofthis environmental pollution. Given that there are numerous EDCs that cannegatively affect humans and wildlife, general screening techniques likebioassays and biosensors provide an essential rapid and intensiveanalysis capacity. Commonly applied bioassays include the ELISA and YESassays, but promising technologies include ER-CALUXa, ELRA, Endotecta,RIANA, and IR-bioamplification. Two biosensors, Endotecta and RIANA, arefield portable using non-cellular biological detection strategies.Environmental management of EDCs in water requires integration ofbiosensors and bioassays for monitoring and assessment.

  20. Enhancement of Colorimetric Response of Enzymatic Reactions by Thermally Evaporated Plasmonic Thin Films: Application to Glial Fibrillary Acidic Protein

    PubMed Central

    Abel, Biebele; Kabir, Tabassum S.; Odukoya, Babatunde; Mohammed, Muzaffer; Aslan, Kadir

    2015-01-01

    We report the enhancement of the colorimetric response of horseradish peroxidase (HRP) and alkaline phosphatase (AP) in bioassays by thermally evaporated silver, gold, copper and nickel thin films. In this regard, a model bioassay based on biotin-avidin interactions was employed. Biotin groups and enzymes were introduced to all surfaces using a biotinylated linker molecule and avidin, respectively. The colorimetric response of HRP in the model bioassay carried out on the plasmonic thin films were up to 4.4-fold larger as compared to control samples (i.e., no plasmonic thin films), where the largest enhancement of colorimetric response was observed on silver thin films. The colorimetric response of AP on plasmonic thin films was found to be similar to those observed on control samples, which was attributed to the loss of enzymes from the surface during the bioassay steps. The extent of enzymes immobilized on to plasmonic thin films was found to affect the colorimetric response of the model bioassay. These findings allowed us to demonstrate the use of silver thin films for the detection of glial fibrillary acidic protein (GFAP), where the colorimetric response of the standard bioassays for GFAP was enhanced up to 67% as compared to bioassays on glass slides. PMID:25663850

  1. Laboratory bioassay for assessing the effects of sludge supernatant on plant growth and vesicular-arbuscular mycorrhiza formation

    SciTech Connect

    Bohn, K.S.; Liberta, A.E.

    1982-12-01

    A laboratory bioassay is described for assessing the effects of sludge supernatant on juvenile corn growth and the ability of vesicular-arbuscular (VA) mycorrhizal fungi, indigenous to coal spoil, to form mycorrhizae. The bioassay demonstrated that application rates can be identified that have the potential to promote increased plant dry weight without suppressing the formation of VA mycorrhizae in a plant's root system.

  2. INITIATION/PROMOTION BIOASSAY IN RAT LIVER: USE OF GAMMA GLUTAMYLTRANSPEPTIDASE-POSITIVE FOCI TO INDICATE CARCINOGENIC ACTIVITY

    EPA Science Inventory

    Gamma Glutamyltranspeptidase (GGTase)-positive foci have been used to indicate activity in an initiation/promotion bioassay in rat liver. This rat liver foci bioassay has been proposed for inclusion in tier 2 of a three tier decision tree approach to carcinogenesis testing where ...

  3. Minimization of Between-well Sample Variance of Antifungal Activity Measurements Using a High-Throughput Screening Microplate Bioassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The use of microplate bioassays, or broth microdilution assays, to measure the activity of biological and synthetic compounds against fungal pathogens has increased in recent years; this technique has been identified as the most promising in vitro bioassay for quantifying antifungal activity. Quant...

  4. Evaluating macroinvertebrate population and community level effects in outdoor microcosms: Use of in situ bioassays and multivariate analysis

    SciTech Connect

    Shaw, J.L.; Manning, J.P.

    1996-05-01

    Evaluating toxicant effects on aquatic communities is difficult due to the ecological complexity at higher levels of organization. Two methods were assessed to improve the understanding of effects on macroinvertebrate communities in aquatic model ecosystems. First, in situ bioassay population effects were used to interpret effects at a higher organization level. Second, canonical discriminant analysis was used to investigate effects on community structure. In situ bioassays were conducted on six occasions in 17-m{sup 3} microcosms treated with copper sulfate. Macroinvertebrates occurring naturally in the microcosms were monitored. Epibenthic in situ bioassays were conducted using Caenis sp. (Ephemeroptera) and Hyalella azteca (Amphipoda) and a water column bioassay was conducted using Notonectidae (Hemiptera). Survival and growth were assessed after 3 d. Effects of copper on both notonectidae and Caenis were observed following application. However, the final Caenis epibenthic bioassays indicated that potential for recovery and survival was {ge}95%. Potential for recovery was less distinct in the water column bioassays. Copper effects also occurred on epibenthic macroinvertebrate populations and communities. Only four taxa, including Caenis, distinguished community differences among copper treatments soon after application. Later, communities showed similarities to the pretreatment bioassay. However, actual recovery was less apparent than the potential for recovery indicated by the bioassays, and community differences due to Caenis persisted.

  5. COMPARISON OF BIOASSAY AND ENZYME-LINKED IMMUNOSORBENT ASSAY FOR QUANTIFICATION OF 'SPODOPTERA FRUGIPERDA' NUCLEAR POLYHEDROSIS VIRUS IN SOIL

    EPA Science Inventory

    Standard curves with known amounts of Spodoptera frugiperda nuclear polyhedrosis virus (NPV) in soil were established with a bioassay and with an enzyme-linked immunosorbent assay (ELISA). The bioassay detected as few as 4 x 10 to the 4th power polyhedral inclusion bodies (PIB)/g...

  6. Summary of the EPA (Environmental Protection Agency) workshop on carcinogenesis bioassay via the dermal route. Final technical report

    SciTech Connect

    Not Available

    1987-04-29

    Traditionally, the oral route has been the most common route of administration in bioassays which tested the potential carcinogenicity of chemicals. Regulatory agencies, however, prefer to have test chemicals applied by the same route as expected human exposure, whenever possible. Since human exposure to industrial chemicals is frequently via the dermal route, this has become a route of choice for animal testing of certain chemicals. However, protocol design for dermal bioassays presents many unique problems which must be addressed before guidelines for bioassays by the dermal route can be formulated. Furthermore, it may be feasible to develop a limited dermal protocol to screen certain classes of chemicals such as acrylates/methacrylates. Recognizing the need for this workshop, it was designed in two distinct parts; to address the problems inherent in the development of a generic protocol for dermal bioassays and, a specific limited dermal bioassay protocol for acrylates/methacrylates.

  7. A yeast bioassay for direct measurement of thyroid hormone disrupting effects in water without sample extraction, concentration, or sterilization.

    PubMed

    Li, Jian; Ren, Shujuan; Han, Shaolun; Li, Na

    2014-04-01

    The present study introduces an improved yeast bioassay for rapid yet sensitive evaluation of thyroid hormone disruption at the level of thyroid receptor (TR) in environmental water samples. This assay does not require water sample preparation and thus requires very little hands-on time. Based on different β-galactosidase substrates, two modified bioassays, a colorimetric bioassay and a chemiluminescent bioassay, were developed. The compounds tested included the known thyroid hormone 3,3',5-triiodo-l-thyronine (T3), the specific TR antagonist amiodarone hydrochloride (AH) and phthalate esters (PAEs), which potentially disrupt thyroid hormone signaling. The EC50 values for T3 were similar to those previously obtained using a 96-well plate bioassay. TR antagonism by AH was studied in the presence of 2.5 × 10(-7)M T3, and the concentration producing 20% of the maximum effect (RIC20) for AH was 3.1 × 10(-7)M and 7.8 × 10(-9)M for the colorimetric bioassay and chemiluminescent bioassay, respectively. None of the tested PAEs induced β-galactosidase expression, but diethylhexyl phthalate, benzyl butyl phthalate and dibutyl phthalate demonstrated TR antagonism. Furthermore, water samples collected from Guanting reservoir in Beijing were evaluated. Although TR agonism was not observed, antagonism was detected in all water samples and is expressed as AH equivalents. The toxicology equivalent quantity values obtained by the chemiluminescent bioassay ranged from 21.2 ± 1.6 to 313.9 ± 28.8 μg L(-1) AH, and similar values were obtained for the colorimetric bioassay. The present study shows that the modified yeast bioassay can be used as a valuable tool for quantification of thyroid hormone disrupting effects in environmental water samples. PMID:24355165

  8. OBSERVATIONS ON THE 10-DAY CHIRONOMUS TENTANS SURVIVAL AND GROWTH BIOASSAY IN EVALUATING GREAT LAKES SEDIMENTS

    EPA Science Inventory

    A 10-day bioassay with larval chironomids (Chironomus tentans) was used to evaluate sediment samples from harbors at Michigan City, IN, St. Joseph, MI, Grand Haven, MI and Toledo, OH for toxicity, based on the endpoints of survival, dry weight, and growth. Larval responses in se...

  9. Using a Macroalgal δ15N Bioassay to Detect Cruise Ship Waste Water Effluent Inputs

    EPA Science Inventory

    Nitrogen stable isotopes are a powerful tool for tracking sources of N to marine ecosystems. I used green macroalgae as a bioassay organism to evaluate if the δ15N signature of cruise ship waste water effluent (CSWWE) could be detected in Skagway Harbor, AK. Opportunistic green...

  10. APPLICATION OF SHORT-TERM BIOASSAYS IN THE FRACTIONATION AND ANALYSIS OF COMPLEX ENVIRONMENTAL MIXTURES

    EPA Science Inventory

    The report is the proceedings of a symposium convened at Williamsburg, Virginia February 21-23, 1978. The volume consists of 24 formal presentations that amplify the three major topics discussed during the symposium: an overview of short-term bioassay systems; current methodology...

  11. Relationships of maternal and fetal weight changes in developmental toxicology bioassays

    EPA Science Inventory

    Standard developmental toxicology bioassays are designed to identify agents with the potential to induce adverse effects in the embryo/fetus. Guidelines require the inclusion of a dose level(s) that induces “overt maternal toxicity”. The common occurrence of dose levels at which ...

  12. Maternal and fetal toxicity in developmental toxicology bioassays: Weight changes and their biological significance

    EPA Science Inventory

    Standard developmental toxicology bioassays are designed to identify agents with the potential to induce adverse effects in the embryo/fetus. Guidelines call for the inclusion of a dose level(s) that induces “overt maternal toxicity.” The possibility that general maternal toxicit...

  13. The Intersection of CMOS Microsystems and Upconversion Nanoparticles for Luminescence Bioimaging and Bioassays

    PubMed Central

    Wei, Liping.; Doughan, Samer.; Han, Yi.; DaCosta, Matthew V.; Krull, Ulrich J.; Ho, Derek.

    2014-01-01

    Organic fluorophores and quantum dots are ubiquitous as contrast agents for bio-imaging and as labels in bioassays to enable the detection of biological targets and processes. Upconversion nanoparticles (UCNPs) offer a different set of opportunities as labels in bioassays and for bioimaging. UCNPs are excited at near-infrared (NIR) wavelengths where biological molecules are optically transparent, and their luminesce in the visible and ultraviolet (UV) wavelength range is suitable for detection using complementary metal-oxide-semiconductor (CMOS) technology. These nanoparticles provide multiple sharp emission bands, long lifetimes, tunable emission, high photostability, and low cytotoxicity, which render them particularly useful for bio-imaging applications and multiplexed bioassays. This paper surveys several key concepts surrounding upconversion nanoparticles and the systems that detect and process the corresponding luminescence signals. The principle of photon upconversion, tuning of emission wavelengths, UCNP bioassays, and UCNP time-resolved techniques are described. Electronic readout systems for signal detection and processing suitable for UCNP luminescence using CMOS technology are discussed. This includes recent progress in miniaturized detectors, integrated spectral sensing, and high-precision time-domain circuits. Emphasis is placed on the physical attributes of UCNPs that map strongly to the technical features that CMOS devices excel in delivering, exploring the interoperability between the two technologies. PMID:25211198

  14. Complete Genome Sequence of Cyanobium sp. NIES-981, a Marine Strain Potentially Useful for Ecotoxicological Bioassays

    PubMed Central

    Shimura, Yohei; Suzuki, Shigekatsu; Yamagishi, Takahiro; Tatarazako, Norihisa; Kawachi, Masanobu

    2016-01-01

    Cyanobium sp. NIES-981 is a marine cyanobacterium isolated from tidal flat sands in Okinawa, Japan. Here, we report the complete 3.0-Mbp genome sequence of NIES-981, which is composed of a single chromosome, and its annotation. This sequence information may provide a basis for developing an ecotoxicological bioassay using this strain. PMID:27469961

  15. Utilization of a duckweed bioassay to evaluate leaching of heavy metals in smelter contaminated soils

    SciTech Connect

    Youngman, A.L.; Lydy, M.J.; Williams, T.L.

    1998-12-31

    The purpose of this study was to determine whether a duckweed bioassay could be used to evaluate the downward migration of heavy metals in smelter soils. The duckweed bioassay was initially used to evaluate elutriates prepared from samples of smelter soils. These initial tests verified that the elutriates would elicit toxic responses. Elutriate testing was followed with an evaluation of leachate from untreated soil cores or soil cores that had been amended with organic matter either unplanted or planted to a grass-forb seed mixture. There was an inverse linear relationship between heavy-metal concentrations in leachate and NOEC and IC{sub 50} values expressed as percentages among all soil cores. Based on these preliminary duckweed bioassays, there were no differences between soil types or organic amended or non-amended soil, but leachate from vegetated soil cores were less toxic than were leachates from non-vegetated soil cores. Overall, the duckweed bioassays were useful in detecting heavy metal availability in elutriate and leachate samples from smelter soils.

  16. CADMIUM SPECIATION IN AQUATIC-LIFE FLOW-THROUGH BIOASSAY DILUTERS

    EPA Science Inventory

    A framework of cadmium speciation studies is described for flow-through mini-diluter bioassay systems which employs Lake Superior water containing added concentrations of calcium carbonate, Aldrich humic acid and/or less than 2 microns suspended Lake Superior red clay. Aspects of...

  17. Development of bioassay techniques with extracts from semi-permeable membrane devices (SPMDs)

    SciTech Connect

    Metcalfe, T.L.; White, P.; Mackay, D.; Metcalfe, C.

    1995-12-31

    Semi-permeable membrane devices (SPMDs), consisting of polyethylene bags filled with triolein, have been used to monitor for lipophilic organic contaminants in water. Although extracts from SPMDs have most often been analyzed for concentrations of organic contaminants, there is also the potential to monitor the toxicity of these extracts using in vitro and in vivo bioassays. SPMDs were deployed for four weeks at several sites along a corridor extending from Peche Island in the Detroit River to Pelee Island in western Lake Erie to monitor the distribution of toxic organic contaminants in the water. Analysis of the extracts from the SPMDs for concentrations of PCBs and other organochlorine compounds, and polynuclear aromatic hydrocarbons (PAHs) indicated that the regions in the Detroit River within the Trenton Channel and near Zug Island were the most highly contaminated. Bioassays conducted with extracts from the SPMDs included the in vitro SOS Chromotest for genotoxic activity, an acute lethality test with Daphnia magna, and a fish embryotoxicity test with embryos of Japanese medaka (Oryzias latipes). These bioassay data generally indicated that the toxicity and concentrations of organic contaminants in the SPMD extracts were correlated. This study indicates that there is potential to use short-term bioassays of extracts from SPMDs to monitor for in situ contamination in the aquatic environment.

  18. Development of an in vitro rainbow trout cell bioassay for PCBs and dioxins

    SciTech Connect

    Richter, C.A.; Leykam, V.; Giesy, J.P.; Denison, M.S.

    1994-12-31

    The toxicity of PCBs, dioxins, and other halogenated aromatic hydrocarbons (HAHS) is difficult to predict from chemical analysis alone, since these chemicals occur in complex mixtures and participate in interactions. Since HAH toxicity is correlated with affinity for the aromatic hydrocarbon receptor (AhR), bioassays which measure induction of genes controlled by the AhR can predict toxicity. The authors have developed a bioassay relevant to fish derived from the RTH-149 rainbow trout hepatoma cell line. The parent cell line has been stably transfected with the plasmid pGudLucl.l. This plasmid contains the firefly luciferase reporter gene under the control of mouse DRES. The reporter gene is controlled only by the AhR, and its induction can be detected at low levels. The bioassay will be fully characterized and calibrated to the responses of whole fish exposed to HAHS. The bioassay exhibits detectable induction at 100 fM TCDD after a three day exposure. Induction increases in a linear relationship with time up to four days of exposure. Luciferase activity was induced to 1 7.5 times background activity after four days of exposure to 1 nM TCDD.

  19. The use of bioassay to determine the effects of cooking on the toxicity of fumonisins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fumonisins are mycotoxin contaminants of maize. Fumonisin B1 (FB1), the most common and toxic fumonisin, causes species-specific diseases in animals, is carcinogenic to rodents, and induces neural tube defects (NTD) in LM/Bc and CD1 mouse bioassays. The human health implications associated with FB1...

  20. Determination of Biochemical Oxygen Demand of Area Waters: A Bioassay Procedure for Environmental Monitoring

    ERIC Educational Resources Information Center

    Riehl, Matthew

    2012-01-01

    A graphical method for determining the 5-day biochemical oxygen demand (BOD5) for a body of water is described. In this bioassay, students collect a sample of water from a designated site, transport it to the laboratory, and evaluate the amount of oxygen consumed by naturally occurring bacteria during a 5-day incubation period. An accuracy check,…

  1. Study of the interactions between copper, cadmium, and ferbam using the protozoan Colpidium campylum bioassay.

    PubMed

    Sekkat, N; Le Dû, A; Jouany, J M; Guerbet, M

    1992-12-01

    The toxicity of a copper-cadmium-ferbam mixture has been studied using the protozoan Colpidium campylum bioassay. The assays were designed according to the factorial experiments method, associated with multiple regression analysis. The results show that, at the concentrations tested, a synergy occurs between cadmium and ferbam, whereas the copper is only oligodynamic. PMID:1282874

  2. Experience with NQA-1 quality assurance standards applied to in vitro bioassay

    SciTech Connect

    Bihl, D.E.; MacLellan, J.A.

    1991-10-01

    On June 1, 1990, the large (about 4000 samples per year) excreta bioassay program at the Hanford Site ceased abruptly when the contract with the bioassay laboratory was terminated. An intense, high-priority effort was begun to replace the services on an interim basis until a new contract could be procured. Despite the urgency to get the excreta bioassay program going again, the Hanford Internal Dosimetry Program was constrained to use only labs that could meet stringent quality assurance (QA) requirements, even during the interim period. The QA requirements were based on NQA-1 with selected additions from the Environmental Protection Agency's QAMS 005/80 (EPA 1983) and the American Society for Testing and Materials' C 1009-83 (ASTM 1984). This constraint was driven both by legal reasons and by the Hanford Site contractors and workers not wanting the quality of the data to be sacrificed. Finding labs that could (1) handle the large throughput, (2) meet the technical requirements, and (3) pass the QA audit proved more difficult than first anticipated. This presentation focuses on the QA requirements that the labs had to meet and how those very broad requirements were applied specifically to excreta bioassay. 5 refs.

  3. Complete Genome Sequence of Cyanobium sp. NIES-981, a Marine Strain Potentially Useful for Ecotoxicological Bioassays.

    PubMed

    Yamaguchi, Haruyo; Shimura, Yohei; Suzuki, Shigekatsu; Yamagishi, Takahiro; Tatarazako, Norihisa; Kawachi, Masanobu

    2016-01-01

    Cyanobium sp. NIES-981 is a marine cyanobacterium isolated from tidal flat sands in Okinawa, Japan. Here, we report the complete 3.0-Mbp genome sequence of NIES-981, which is composed of a single chromosome, and its annotation. This sequence information may provide a basis for developing an ecotoxicological bioassay using this strain. PMID:27469961

  4. Is there a role for estrogen activity assays? Recombinant cell bioassay for estrogen: Development and applications.

    PubMed

    Klein, Karen Oerter

    2015-07-01

    There are many questions which cannot be answered without a very sensitive estradiol assay. A recombinant cell bioassay (RCBA) for estradiol was developed in 1994. The sensitivity of the bioassay is 0.02-0.2 pg/ml (0.07-0.7 pmol/L), more than 20 times more sensitive than commercial RIAs and 10 times more sensitive than newer mass spectrometry assays. The RCBA for estradiol opened the door to study low levels of estradiol equivalents (EE) across the physiological spectrum of life from prepubertal children through menopause and across the spectrum from normal physiology, in boys as well as girls, to pathology, including: premature thelarche; estradiol suppression in children treated with GnRH analogues for precocious puberty; aromatase inhibition in boys with growth hormone deficiency; the differences between oral and transdermal routes of estrogen administration in girls with Turner's syndrome; women with breast cancer treated with aromatase inhibitors; and women with urogenital atrophy treated with low dose vaginal estrogen. A bioassay also allows study of endocrine disruptors, like phytoestrogens and other environmental compounds, which are relevant to public health and alternative medicine options. This paper reviews the assay and the last 20 years of applications. A bioassay for estrogen has a role because measuring biological effect is theoretically useful, increasing the understanding of physiology in addition to biochemical levels, giving different information than other assays, and opening the door to measure very low levels of estrogen activity in both humans and the environment. PMID:25159103

  5. A cellulose-based bioassay for the colorimetric detection of pathogen DNA.

    PubMed

    Saikrishnan, Deepika; Goyal, Madhu; Rossiter, Sharon; Kukol, Andreas

    2014-12-01

    Cellulose-paper-based colorimetric bioassays may be used at the point of sampling without sophisticated equipment. This study reports the development of a colorimetric bioassay based on cellulose that can detect pathogen DNA. The detection was based on covalently attached single-stranded DNA probes and visual analysis. A cellulose surface functionalized with tosyl groups was prepared by the N,N-dimethylacetamide-lithium chloride method. Tosylation of cellulose was confirmed by scanning electron microscopy, Fourier transform infrared spectroscopy and elemental analysis. Sulfhydryl-modified oligonucleotide probes complementary to a segment of the DNA sequence IS6110 of Mycobacterium tuberculosis were covalently immobilized on the tosylated cellulose. On hybridization of biotin-labelled DNA oligonucleotides with these probes, a colorimetric signal was obtained with streptavidin-conjugated horseradish peroxidase catalysing the oxidation of tetramethylbenzamidine by H2O2. The colour intensity was significantly reduced when the bioassay was subjected to DNA oligonucleotide of randomized base composition. Initial experiments have shown a sensitivity of 0.1 μM. A high probe immobilization efficiency (more than 90 %) was observed with a detection limit of 0.1 μM, corresponding to an absolute amount of 10 pmol. The detection of M. tuberculosis DNA was demonstrated using this technique coupled with PCR for biotinylation of the DNA. This work shows the potential use of tosylated cellulose as the basis for point-of-sampling bioassays. PMID:25354892

  6. Bioassay and Attributes of a Growth Factor Associated with Crown Gall Tumors 1

    PubMed Central

    Lippincott, Barbara B.; Lippincott, James A.

    1970-01-01

    An improved bioassay is described for a factor that promotes tumor growth which was first obtained from extracts of pinto bean leaves with crown gall tumors. Sixteen primary pinto bean leaves per sample are inoculated with sufficient Agrobacterium tumefaciens to initiate about 5 to 10 tumors per leaf and treated with tumor growth factor at day 3 after inoculation. The diameters of 30 to 48 round tumors (no more than 3 randomly selected per leaf) are measured per test sample at day 6. Mean tumor diameter increased linearly with the logarithm of the concentration of tumor growth factor applied. The tumor growth factor was separated by column chromatography from an ultraviolet light-absorbing compound previously reported to be associated with fractions having maximal tumor growth factor activity. Partly purified tumor growth factor showed no activity in a cytokinin bioassay or an auxin bioassay, and negligible activity in gibberellin bioassays. Representatives of these three classes of growth factors did not promote tumor growth. Extracts from crown gall tumors on primary pinto bean leaves, primary castor bean leaves, Bryophyllum leaves, carrot root slices, and tobacco stems showed tumor growth factor activity, whereas extracts from healthy control tissues did not. Extracts from actively growing parts of healthy pinto beans, Bryophyllum, and tobacco, however, showed tumor growth factor activity. Tumor growth factor is proposed to be a normal plant growth factor associated with rapidly growing tissues. Its synthesis may be activated in nongrowing tissues by infection with Agrobacterium sp. PMID:16657534

  7. SHORT-TERM CARCINOGENESIS AND MUTAGENESIS BIOASSAYS OF UNREGULATED AUTOMOTIVE EMISSIONS

    EPA Science Inventory

    Evaluation of the potential risk of a chemical or environmental emission causing chronic health effects requires data from one or more of the following sources: epidemiologic and clinical studies of human exposure and effects; chronic (long term) bioassays in animals; and short-t...

  8. TECHNIQUE FOR REMOVAL OF DISSOLVED AND DISPERSED HYDROCARBONS FROM BIOASSAY EFFLUENTS

    EPA Science Inventory

    A method for the efficient removal of petroleum-derived hydrocarbons from the oil-contaminated effluent of a continuous flow-through oil bioassay system is described. The concentration of No. 2 fuel oil in the effluent, discharged at rates from 17 to 26 L/min, is reduced from an ...

  9. Evaluation of soil bioassays for use at Washington state hazardous waste sites: A pilot study

    SciTech Connect

    Blakley, N.; Norton, D.; Stinson, M.; Boyer, R.

    1994-12-31

    The Washington State Department of Ecology (Ecology) is developing guidelines to assess soil toxicity at hazardous waste sites being investigated under the Washington Model Toxics Control Act Cleanup Regulation. To evaluate soil toxicity, Ecology selected five bioassay protocols -- Daphnia, Earthworm, Seedling, Fathead Minnow, and Frog Embryo Teratogenesis Assay Xenopus (FETAX) -- for use as screening level assessment tools at six State hazardous waste sites. Sites contained a variety of contaminants including metals, creosote, pesticides, and petroleum products (leaking underground storage tanks). Three locations, representing high, medium, and low levels of contamination, were samples at each site. In general, the high contaminant samples resulted in the highest toxic response in all bioassays. The order of site toxicity, as assessed by overall toxic response, is creosote, petroleum products, metals, and pesticides. Results indicate that human health standards, especially for metals, may not adequately protect some of the species tested. The FETAX bioassay had the greatest overall number of toxic responses and lowest variance. The seedling and Daphnia bioassays had lower and similar overall toxic response results, followed by the earthworm and fathead minnow. Variability was markedly highest for the seedling. The Daphnia and fathead minnow variability were similar to the FETAX level, while the earthworm variability was slightly higher.

  10. An Estuarine Fish Bioassay for Sensitive Biomonitoring of Oil-related Contamination

    EPA Science Inventory

    An embryonic and larval bioassay using the estuarine fish, Fundulus heteroclitus, was modified for the rapid detection of bioavailable compounds that act through the aryl hydrocarbon receptor (AhR). The early development of fishes is particularly sensitive to AhR agonists, such ...

  11. SUPERNUMERARY RIBS IN DEVELOPMENTAL TOXICITY BIOASSAYS AND IN HUMAN POPULATIONS: INCIDENCE AND BIOLOGICAL SIGNIFICANCE

    EPA Science Inventory

    Abstract
    Supernumerary or accessory ribs (SNR), either lumbar (LSNR) or cervical (CSNR) are a common finding in standard developmental toxicology bioassays. The biological significance of these anomalies within the regulatory arena has been problematic and the subject of some...

  12. Olfactoryresponse of the predatory mite Typhlodromus pyri (Acari: Phytoseiidae) to methyl salicylate in laboratory bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The response of Typhlodromus pyri, a key predator of grapevine rust mite (Calepitrimerus vitis), to MeSA was tested using a Y-tube olfactometer in laboratory bioassays. Six doses ranging from 200 to 0.002 µg of diluted MeSA were tested. Significantly higher proportions of T. pyri preferred MeSA at ...

  13. Determining UV Inactivation of Toxoplasma gondii Oocysts by Using Cell Culture and a Mouse Bioassay

    EPA Science Inventory

    The effect of UV exposure on Toxoplasma gondii oocysts has not been completely defined for use in water disinfection. This study evaluated UV irradiated oocysts by three assays: a SCID mouse bioassay, an in vitro T. gondii oocyst plaque assay (TOP-assay), and a quantitative reve...

  14. Statistical method for determining and comparing limits of detection of bioassays.

    PubMed

    Holstein, Carly A; Griffin, Maryclare; Hong, Jing; Sampson, Paul D

    2015-10-01

    The current bioassay development literature lacks the use of statistically robust methods for calculating the limit of detection of a given assay. Instead, researchers often employ simple methods that provide a rough estimate of the limit of detection, often without a measure of the confidence in the estimate. This scarcity of robust methods is likely due to a realistic preference for simple and accessible methods and to a lack of such methods that have reduced the concepts of limit of detection theory to practice for the specific application of bioassays. Here, we have developed a method for determining limits of detection for bioassays that is statistically robust and reduced to practice in a clear and accessible manner geared at researchers, not statisticians. This method utilizes a four-parameter logistic curve fit to translate signal intensity to analyte concentration, which is a curve that is commonly employed in quantitative bioassays. This method generates a 95% confidence interval of the limit of detection estimate to provide a measure of uncertainty and a means by which to compare the analytical sensitivities of different assays statistically. We have demonstrated this method using real data from the development of a paper-based influenza assay in our laboratory to illustrate the steps and features of the method. Using this method, assay developers can calculate statistically valid limits of detection and compare these values for different assays to determine when a change to the assay design results in a statistically significant improvement in analytical sensitivity. PMID:26376354

  15. A MARINE ALGAL BIOASSAY METHOD: RESULTS WITH PESTICIDES AND INDUSTRIAL WASTES

    EPA Science Inventory

    A simple marine algal bioassay method is described for short- and long-term studies on pesticides and industrial wastes. It can be used for rapid screening of a variety of substances with single-species and multiple-species tests and gives relative toxicities of the pollutants te...

  16. A SIMPLE, RAPID BIOASSAY FOR DETECTING EFFECTS OF POLLUTANTS ON BACTERIA

    EPA Science Inventory

    Since approximately 90% of hazardous wastes reach soil and water for permanent disposal, it is logical that microflora (bacteria) contained in these environments be used to establish initial toxicity levels. Bacteria can be suitable bioassay tools because they are inexpensive to ...

  17. Risk assessment for selected xenobiotics by bioassay methods with higher plants

    NASA Astrophysics Data System (ADS)

    Günther, Petra; Pestemer, Wilfried

    1990-05-01

    Different bioassays with higher plants were approved for use in a bioassay procedure for testing of xenobiotics according to the German Chemicals Act. Selected environmental pollutants (atrazine, cadmium chloride, 2,6-dichlorobenzonitrile, pentachlorophenol, potassium dichromate, thiourea), all from a list of reference chemicals, were tested with these methods. Dose-response curves for growth of oats and turnips were evaluated in soil and vermiculite (nonsorptive substrate), and availability to plants was calculated by comparing the EC50 values for one chemical in both substrates. The most active chemical was atrazine, followed by 2,6-dichlorobenzonitrile, pentachlorophenol, potassium dichromate, cadmium chloride, and thiourea. The least available compound to plants was pentachlorophenol, tested with turnips ( Brassica rapa var. rapa). The strongest inhibition of germination, demonstrated in an in vitro assay with garden cress ( Lepidium sativum), was found with 2,6-dichlorobenzonitrile, the lowest with atrazine. The effect of an extended exposure of the plants to the chemicals was evaluated in a long-term bioassay with oats ( Avena sativa) in hydroponic culture. Several dose-response curves during the growing period were derived. It was found that the EC50 values for atrazine and thiourea decreased markedly during the first four weeks; thereafter the changes were much smaller. As an overall conclusion, a bioassay procedure is proposed that can be included in the graduated plan recommended by the German Chemicals Act.

  18. Rapid diagnosis of imazapic & imazapyr resistance by using bioassays in Clearfield Production System, Malaysia

    NASA Astrophysics Data System (ADS)

    Bajrai F. S., M.; Ismail B., S.; Mardiana-Jansar, Khairiatul

    2015-09-01

    The resistance of weedy rice biotypes toward OnDuty™WG has been reported in Clearfield® MR 220 CL1 and MR 220 CL2 types of paddy. The purpose of this study was to adopt a rapid method to evaluate the resistance of bioassay species towards imazapic + imazapyr in different stages of plant development (seeds and seedlings). A series of OnDuty™WG concentrations from 0 to 300 g ai ha-1 were studied on the growth of rice cultivar MR263 (a susceptible species) as the bioassay species. The experiments were done in three replications with Complete Randomized Block Design (CRBD). From this study, the concentration of herbicide required to reduce coleoptiles length, root length and fresh weight in seed bioassay by 50% were 0.63, 0.33 and 3.60 g ai ha-1 respectively. Meanwhile, for seedling stage bioassay, the concentration of herbicide required to reduce coleoptiles length, root length and fresh weight by 50% were 0.03, 1.23 and 0.99 g ai ha-1 respectively. It is important to note that all growth parameters were concentration dependent and a total growth inhibition occurred in all parameters at high doses. It was proven that MR263 rice cultivar was not resistance towards imazapic + imazapyr and further experiments on other rice cultivars are recommended so that the most suitable cultivars will be selected in rice cultivation.

  19. Utilizing high throughput bioassays to characterize the bioactivity of complex environmental samples

    EPA Science Inventory

    Bioassays can be employed to evaluate the integrated effects of complex mixtures of both known and unidentified contaminants present in environmental samples. However, such methods have typically focused on one or a few bioactivities despite the fact that the chemicals in a mixtu...

  20. Efficiency of several cultural methods and a chick bioassay to recover dry stressed Campylobacter

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aims of the study were to evaluate the efficacy of 5 enrichment procedures for recovery of dry-atmospheric-temperature stressed C. jejuni and C. coli and determine the viable status of the non-culturable strains using a chick bioassay. Sterile chick paper pads (PP) and filter papers (FP) were i...

  1. The release of rat intestinal cholecystokinin after oral trypsin inhibitor measured by bio-assay.

    PubMed Central

    Brand, S J; Morgan, R G

    1981-01-01

    The distribution, molecular form and release of cholecystokinin (CCK)-like activity in extracts of rat small intestine was studied with an in vitro gall-bladder bio-assay. In contrast to the reported heterogeneity of CCK-like immunoreactivity in the intestine, only a single molecular form of CCK-like activity was detected using the bio-assay. 2. The CCK-like activity eluted from Sephadex G50 with a Kav of 0.69, after the triacontriapeptide of cholecystokinin (CCK33) and before cholecystokinin octapeptide 2500, may represent the 22 amino acid peptide of CCK (CCK22). The bio-assay peak of CCK-like activity had pancreozymin activity and CCK/gastrin C terminal immunoreactivity. The CCK-like activity weas readily extracted from the small intestine at neutral pH, but subsequent treatment with cold 0.5 M-acetic acid extracted further CCK-like activity of the same molecular form as that recovered under neutral conditions. 3. The bio-assay detected no CCK-like activity, nor was pancreozymin-like activity found in fractions corresponding to CCK33 or CCK8 after Sephadex G50 chromatography of rat intestinal extracts. 4. Oral trypsin inhibitor was a potent stimulus for the release of CCK-like activity from the upper small intestine of the rat. After oral trypsin inhibitor release, CCK-like activity was rapidly resynthesized. PMID:7320918

  2. A bioassay approach for determining the effect of cooking on fumonisin today

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fumonisin mycotoxins are found in corn and corn-based foods, but the effect of cooking on fumonisin toxicity has not been studied extensively. Rat feeding studies were used as an in vivo bioassay to compare the toxicity of extrusion cooked and nixtamalized (alkaline cooked) fumonisin-contaminated p...

  3. Parlin, a general microcomputer program for parallel-line analysis of bioassays.

    PubMed

    Jesty, J; Godfrey, H P

    1986-04-01

    Commonly used manual and calculator methods for analysis of clinically important parallel-line bioassays are subject to operator bias and provide neither confidence limits for the results nor any indication of their validity. To remedy this, the authors have written a general program for statistical analysis of these bioassays for the IBM Personal Computer and its compatibles. The program has been used for analysis of bioassays for specific coagulation factors and inflammatory lymphokines and for radioimmunoassays for prostaglandins. The program offers a choice of no transform, logarithmic, or logit transformation of data, which are fitted to parallel lines for standard and unknown. It analyzes the fit for parallelism and linearity with an F test, and calculates the best estimate of the result and its 95% confidence limits. Comparison of results calculated by PARLIN with those previously obtained manually shows excellent correlation (r greater than 0.99). Results obtained using PARLIN are quickly available with current assay technics and provide a complete evaluation of the bioassay at no increase in cost. PMID:3456698

  4. Novel bacterial bioassay for a high-throughput screening of 4-hydroxyphenylpyruvate dioxygenase inhibitors.

    PubMed

    Rocaboy-Faquet, Emilie; Noguer, Thierry; Romdhane, Sana; Bertrand, Cédric; Dayan, Franck Emmanuel; Barthelmebs, Lise

    2014-08-01

    Plant 4-hydroxyphenylpyruvate dioxygenase (HPPD) is the molecular target of a range of synthetic β-triketone herbicides that are currently used commercially. Their mode of action is based on an irreversible inhibition of HPPD. Therefore, this inhibitory capacity was used to develop a whole-cell colorimetric bioassay with a recombinant Escherichia coli expressing a plant HPPD for the herbicide analysis of β-triketones. The principle of the bioassay is based on the ability of the recombinant E. coli clone to produce a soluble melanin-like pigment, from tyrosine catabolism through p-hydroxyphenylpyruvate and homogentisate. The addition of sulcotrione, a HPPD inhibitor, decreased the pigment production. With the aim to optimize the assay, the E. coli recombinant clone was immobilized in sol-gel or agarose matrix in a 96-well microplate format. The limit of detection for mesotrione, tembotrione, sulcotrione, and leptospermone was 0.069, 0.051, 0.038, and 20 μM, respectively, allowing to validate the whole-cell colorimetric bioassay as a simple and cost-effective alternative tool for laboratory use. The bioassay results from sulcotrione-spiked soil samples were confirmed with high-performance liquid chromatography. PMID:24816780

  5. A RAPID, QUANTITATIVE BIOASSAY FOR DETECTING PHYTOTOXIC GASES USING STRESS-ETHYLENE

    EPA Science Inventory

    A simple bioassay for detecting phytotoxic air pollutants has been developed. Wheat (Triticum aestivum L.) seedlings growing on agar medium in test-tubes are exposed to pollutants for 2 h. Stress-ethylene produced by the seedlings during exposure to the pollutants is collected in...

  6. DEVELOPMENT OF A TEST-TUBE STRESS-ETHYLENE BIOASSAY FOR DETECTING PHYTOTOXIC GASES

    EPA Science Inventory

    A rapid, quantitative bioassay for detecting phytotoxic air pollutants has been developed. The technique uses wheat Triticum aestivum L. or tomato Lycopersicon esculentum L., seedlings growing on an agar medium in test-tubes. The seedlings are exposed to a pollutant in the test-t...

  7. USE OF SALMONELLA MICROSUSPENSION BIOASSAY TO DETECT THE MUTGENICITY OF MUNITIONS COMPOUNDS AT LOW CONCENTRATIONS

    EPA Science Inventory



    Use of a Salmonella Microsuspension Bioassay to Detect the Mutagenicity of
    Munitions Compounds at Low Concentrations

    Abstract

    Past production and handling of munitions has resulted in soil contamination at various military facilities. Depending on...

  8. Age and sex related responsiveness of Tribolium castaneum (Coleoptera: Tenebrionidae) in novel behavioral bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The hardiness and mobile nature of Tribolium castaneum (Herbst) make them easy to work with but are the same factors that make their responses to behavior modifying chemical stimuli difficult to evaluate. To overcome these difficulties we developed two bioassays: a two choice test with airflow and a...

  9. Effective bioassays for evaluating boxwood blight (Calonectria pseudonaviculata) susceptibility using detached stem inoculations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two simple and rapid in vitro bioassays using detached stems were developed for evaluating the susceptibility of boxwood genotypes to the blight disease caused by Calonectria pseudonaviculata. Individual leaves were inoculated on detached stems or entire detached stems were sprayed to assess suscept...

  10. Kinetic microplate bioassays for relative potency of antibiotics improved by partial Least Square (PLS) regression.

    PubMed

    Francisco, Fabiane Lacerda; Saviano, Alessandro Morais; Almeida, Túlia de Souza Botelho; Lourenço, Felipe Rebello

    2016-05-01

    Microbiological assays are widely used to estimate the relative potencies of antibiotics in order to guarantee the efficacy, safety, and quality of drug products. Despite of the advantages of turbidimetric bioassays when compared to other methods, it has limitations concerning the linearity and range of the dose-response curve determination. Here, we proposed to use partial least squares (PLS) regression to solve these limitations and to improve the prediction of relative potencies of antibiotics. Kinetic-reading microplate turbidimetric bioassays for apramacyin and vancomycin were performed using Escherichia coli (ATCC 8739) and Bacillus subtilis (ATCC 6633), respectively. Microbial growths were measured as absorbance up to 180 and 300min for apramycin and vancomycin turbidimetric bioassays, respectively. Conventional dose-response curves (absorbances or area under the microbial growth curve vs. log of antibiotic concentration) showed significant regression, however there were significant deviation of linearity. Thus, they could not be used for relative potency estimations. PLS regression allowed us to construct a predictive model for estimating the relative potencies of apramycin and vancomycin without over-fitting and it improved the linear range of turbidimetric bioassay. In addition, PLS regression provided predictions of relative potencies equivalent to those obtained from agar diffusion official methods. Therefore, we conclude that PLS regression may be used to estimate the relative potencies of antibiotics with significant advantages when compared to conventional dose-response curve determination. PMID:26971814

  11. Comparison of two bioassay methods for determining deltamethrin resistance in German cockroaches (Blattodea: Blattellidae).

    PubMed

    Choo, L E; Tang, C S; Pang, F Y; Ho, S H

    2000-06-01

    Susceptibility to deltamethrin of 10 strains of the German cockroach, Blattella germanica (L.), trapped in hotel kitchens in Singapore was determined. Two resistance detection bioassay methods (topical application and World Health Organization glass jar method) were compared. Resistance ratios based on 50% knockdown, obtained by comparison with the S strain, ranged from 17.7 to 4,235 for topical application and from 2.2 to 22 for the glass jar method. A field strain, with consistently low resistance ratios (topical method = 17.7, glass jar method = 2.2), was identified as a potential field strain to be used as a baseline for comparison with other field strains. Resistance ratios for the other field strains obtained by comparison with the R5 strain ranged from 24.5 to 239 for topical application and from 1.2 to 9.8 for the glass jar method. The results of our study demonstrate that deltamethrin-resistant German cockroaches are numerous in Singapore. Comparison between the two bioassay methods showed that there was significant correlation between KD50 and KT50 values. The glass jar method is similar to field situations but topical application is sensitive enough to define the magnitude of resistance. Differences between the two detection bioassays and the factors governing the choice of bioassay in monitoring resistance in German cockroaches are discussed. PMID:10902348

  12. Rapid Bioassessment and In Situ Bioassay: Cost Effective Tools for Environmental Impact Assessment

    SciTech Connect

    Wike, L.D.

    2002-08-23

    Environmental impact can be difficult to assess, especially at the ecosystem level. Any impact assessment methodology that can give cost effective and timely results is highly desirable. Rapid bioassessment (RBA) is cost effective and produces timely results. Several types of RBA have been used at the Savannah River Site (SRS) to assess stream conditions, including the Index of Biotic Integrity (IBI) based on fish community characteristics, and various techniques using aquatic macroinvertebrate species diversity and abundance. In an attempt to broaden the applicability of the RBA concept, we have also begun to develop RBA techniques for seep-fed wetlands and terrestrial habitats. These techniques will focus on vertebrate and macroinvertebrate assemblages for seep-fed wetlands and arthropod assemblages for terrestrial habitats. In situ bioassay is another technique that could be used for rapid and economical assessment of the effects of anthropogenic disturbance. We propose the development of two methods of in situ bioassay that can address bioavailability of constituents of concern. The use of caged bioassay organisms can be applied to terrestrial systems such as capped or existing waste sites using the common house cricket. Another proposed bioassay could use a resident species, such as the imported red fire ant, which is found in disturbed habitats and open areas such as waste sites. Combining in situ techniques with RBA methodologies has the potential to provide a comprehensive assessment of chemical and physical impacts to a wide range of ecosystem types.

  13. APPLICATION OF PLANT AND EARTHWORM BIOASSAYS TO EVALUATE REMEDIATION OF A LEAD-CONTAMINATED SOIL

    EPA Science Inventory

    Earthworm acute toxicity, plant seed germination/root elongation (SG/RE) and plant genotoxicity bioassays were employed to evaluate the remediation of a lead-contaminated soil. The remediation involved removal of heavy metals by a soil washing/soil leaching treatment process. A p...

  14. A BIOASSAY THAT IDENTIFIES POSTNATAL FUNCTIONAL DEFICITS IN MICE PRENATALLY EXPOSED TO XENOBIOTICS

    EPA Science Inventory

    Experimental strategies to evaluate adverse postnatal effects due to prenatal exposure exist for many organ systems. Often, however, there is insufficient information to suggest that a particular organ system(s) may be sensitive to the test agent. A single bioassay to identify ...

  15. PET/CT Imaging of c-Myc Transgenic Mice Identifies the Genotoxic N-Nitroso-Diethylamine as Carcinogen in a Short-Term Cancer Bioassay

    PubMed Central

    Laenger, Florian; Halter, Roman; Rodt, Thomas; Galanski, Michael; Borlak, Juergen

    2012-01-01

    Background More than 100,000 chemicals are in use but have not been tested for their safety. To overcome limitations in the cancer bioassay several alternative testing strategies are explored. The inability to monitor non-invasively onset and progression of disease limits, however, the value of current testing strategies. Here, we report the application of in vivo imaging to a c-Myc transgenic mouse model of liver cancer for the development of a short-term cancer bioassay. Methodology/Principal Findings μCT and 18F-FDG μPET were used to detect and quantify tumor lesions after treatment with the genotoxic carcinogen NDEA, the tumor promoting agent BHT or the hepatotoxin paracetamol. Tumor growth was investigated between the ages of 4 to 8.5 months and contrast-enhanced μCT imaging detected liver lesions as well as metastatic spread with high sensitivity and accuracy as confirmed by histopathology. Significant differences in the onset of tumor growth, tumor load and glucose metabolism were observed when the NDEA treatment group was compared with any of the other treatment groups. NDEA treatment of c-Myc transgenic mice significantly accelerated tumor growth and caused metastatic spread of HCC in to lung but this treatment also induced primary lung cancer growth. In contrast, BHT and paracetamol did not promote hepatocarcinogenesis. Conclusions/Significance The present study evidences the accuracy of in vivo imaging in defining tumor growth, tumor load, lesion number and metastatic spread. Consequently, the application of in vivo imaging techniques to transgenic animal models may possibly enable short-term cancer bioassays to significantly improve hazard identification and follow-up examinations of different organs by non-invasive methods. PMID:22319569

  16. Paper-based chromatic toxicity bioassay by analysis of bacterial ferricyanide reduction.

    PubMed

    Pujol-Vila, F; Vigués, N; Guerrero-Navarro, A; Jiménez, S; Gómez, D; Fernández, M; Bori, J; Vallès, B; Riva, M C; Muñoz-Berbel, X; Mas, J

    2016-03-01

    Water quality assessment requires a continuous and strict analysis of samples to guarantee compliance with established standards. Nowadays, the increasing number of pollutants and their synergistic effects lead to the development general toxicity bioassays capable to analyse water pollution as a whole. Current general toxicity methods, e.g. Microtox(®), rely on long operation protocols, the use of complex and expensive instrumentation and sample pre-treatment, which should be transported to the laboratory for analysis. These requirements delay sample analysis and hence, the response to avoid an environmental catastrophe. In an attempt to solve it, a fast (15 min) and low-cost toxicity bioassay based on the chromatic changes associated to bacterial ferricyanide reduction is here presented. E. coli cells (used as model bacteria) were stably trapped on low-cost paper matrices (cellulose-based paper discs, PDs) and remained viable for long times (1 month at -20 °C). Apart from bacterial carrier, paper matrices also acted as a fluidic element, allowing fluid management without the need of external pumps. Bioassay evaluation was performed using copper as model toxic agent. Chromatic changes associated to bacterial ferricyanide reduction were determined by three different transduction methods, i.e. (i) optical reflectometry (as reference method), (ii) image analysis and (iii) visual inspection. In all cases, bioassay results (in terms of half maximal effective concentrations, EC50) were in agreement with already reported data, confirming the good performance of the bioassay. The validation of the bioassay was performed by analysis of real samples from natural sources, which were analysed and compared with a reference method (i.e. Microtox). Obtained results showed agreement for about 70% of toxic samples and 80% of non-toxic samples, which may validate the use of this simple and quick protocol in the determination of general toxicity. The minimum instrumentation

  17. EFFECT OF THE OIL DISPERSANT OMNI-CLEAN(R) ON THE TOXICITY OF FUEL OIL NO. 2 IN TWO BIOASSAYS WITH THE SHEEPSHEAD MINNOW CYPRINODON VARIEGATUS

    EPA Science Inventory

    Bioassays (7-day early life stage and 96h acute bioassays) were conducted with the sheepshead minnow, Cyprinodon variegatus, to determine the toxicity of the dispersant Omni-Clean< by itself and in combination with fuel oil no. 2. Performance characteristics of both bioassay type...

  18. Responses of lone star tick (acari: ixodidae) nymphs to the repellent deet applied in acetone and ethanol solutions in vitro bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Behavioral bioassays remain a standard tool in the discovery, development, and registration of repellents. Although tick repellent bioassays tend to be rather uncomplicated, several factors can influence their outcomes. Typically repellent bioassays use a solvent, such as acetone or ethanol, to disp...

  19. Electrogenerated Chemiluminescence Bioassay of Two Protein Kinases Incorporating Peptide Phosphorylation and Versatile Probe.

    PubMed

    Liu, Xia; Dong, Manman; Qi, Honglan; Gao, Qiang; Zhang, Chengxiao

    2016-09-01

    A sensitive electrogenerated chemiluminescence (ECL) bioassay was developed for the detection of two protein kinases incorporating the peptide phosphorylation and a versatile ECL probe. Cyclic adenosine monophosphate-dependent protein kinase (PKA) and casein kinase II (CK2) were used as proof-of-concept targets while a PKA-specific peptide (CLRRASLG) and a CK2-specific peptide (CRRRADDSDDDDD) were used as the recognition substrates. Taking advantage of the ability of protein A binding with the Fc region of a variety of antibodies with high affinity, a ruthenium derivative-labeled protein A was utilized as a versatile ECL probe for bioassay of multiple protein kinases. A specific peptide substrate toward target protein kinase was first self-assembled on the surface of gold electrode and then serine in the specific peptide on the electrode was phosphorylated by target protein kinase in the presence of adenosine-5'-triphosphate. After recognition of the phosphorylated peptide by monoclonal antiphosphoserine antibody, the versatile ECL probe was specifically bound to the antiphosphoserine antibody on the electrode surface. The ECL bioassay was developed successfully in the individual detection of PKA and CK2 with detection limit of 0.005 U/mL and 0.004 U/mL, respectively. In addition, the ECL bioassay was applied to quantitative analysis of the kinase inhibitors and monitoring drug-triggered kinase activation in cell lysates. Moreover, an ECL imaging bioassay using electron-multiplying charged coupled device as detector on the gold electrode array was developed for the simultaneous detection of PKA and CK2 activity from 0.01 U/mL to 0.4 U/mL, respectively, at one time. This work demonstrates that the ingenious design and use of a versatile ECL probe are promising to simultaneous detection of multiple protein kinases and screening of kinase inhibitor. PMID:27518533

  20. Bioassays for the detection of chemicals that can form bioactivation-dependent reactive free radicals

    SciTech Connect

    Sanderson, J.T.; Commandeur, J.N.M.; Wezel, A. van; Vermeulen, N.P.E. . Div. of Molecular Toxicology National Inst. for Coastal and Marine Management, Den Haag )

    1999-06-01

    In vitro bioassays were developed for the detection of chemicals that can be bioactivated to reactive free radical species in microsomal fractions. Two methods were deployed, a down-scaled spectrophotometric method for the detection of chemicals that can cause lipid peroxidation using the measurement of thiobarbituric acid-reactive substances (TBARS) and a fluorometric method for the detection of chemicals that can undergo redox cycling to generate superoxide radicals based on the detection of hydrogen peroxide. The response of these systems to prototypical and environmentally relevant chemicals, including tetrachloromethane and paraquat, was examined. The detection limit of the lipid peroxidation bioassay, based on the formation of TBARS, was about 1 [micro]M for tetrachloromethane; that of the bioassay for redox cyclers, based on the production of hydrogen peroxide, was about 2 [micro]M for paraquat and about 100-fold lower for the potent redox cycler 2,3,5,6-tetramethylbenzoquinone (TMBQ). Several binary mixtures of chemicals were tested for potential nonadditive effects in both in vitro systems. Some antagonistic effects among halogenated methanes were observed in the lipid peroxidation assay. In the hydrogen peroxide production assay, greater than additive effects were seen between small concentrations of paraquat and TMBQ. A number of surface water concentrates from several locations in The Netherlands, with various levels of chemical contamination, exhibited a weak response in the hydrogen peroxide production assay. Acetone was found to interfere with the response of the bioassay to redox cyclers and, therefore, the water concentrates (originally in acetone) were transferred to ethanol prior to testing. A good correlation was observed between the response of the water concentrates in the hydrogen peroxide production assay and their acute toxicity in Daphnia magna. No correlation was observed between this bioassay response and toxicity in the Microtox

  1. The OECD program to validate the rat uterotrophic bioassay. Phase 2: dietary phytoestrogen analyses.

    PubMed Central

    Owens, William; Ashby, John; Odum, Jenny; Onyon, Lesley

    2003-01-01

    Many commercial laboratory diets have detectable levels of isoflavones (e.g., phytoestrogens such as genistein [GN]) that have weak estrogenic activity both in vitro and in vivo. During validation studies of the uterotrophic bioassay, diet samples from 20 participating laboratories were collected and analyzed for three major phytoestrogens: GN, daidzein (DN), and coumestrol (CM). Soy phytoestrogens GN and DN were found at total phytoestrogen levels from 100 to 540 microg/g laboratory diet; a forage phytoestrogen, CM, ranged from nondetectable to 4 microg/g laboratory diet. The phytoestrogen levels were compared with both baseline uterine weights of the control groups and with the relative uterine weight increase of groups administered two weak estrogen agonists: bisphenol A (BPA) and nonylphenol (NP). The comparison uses a working assumption of additivity among the phytoestrogens, despite several significant qualifications to this assumption, to estimate total genistein equivalents (TGE). Some evidence was found that phytoestrogen levels in the diet > 325-350 microg/g TGE could diminish the responsiveness of the uterotrophic bioassay to weak agonists. This was especially true for the case of the intact, immature female version of the uterotrophic bioassay, where higher food consumption relative to body weight leads to higher intakes of dietary phytoestrogens versus ovariectomized adults. This dietary level is sufficient in the immature female to approach a biological lowest observable effect level for GN of 40-50 mg/kg/day. These same data, however, show that low to moderate levels of dietary phytoestrogens do not substantially affect the responsiveness of the assay with weak estrogen receptor agonists such as NP and BPA. Therefore, laboratories conducting the uterotrophic bioassay for either research or regulatory purposes may routinely use diets containing levels of phytoestrogens < 325-350 microg/g TGE without impairing the responsiveness of the bioassay. PMID

  2. Scientific Considerations for Evaluating Cancer Bioassays Conducted by the Ramazzini Institute

    PubMed Central

    Caldwell, Jane C.; Jinot, Jennifer; Evans, Marina V.; Cote, Ila; Vandenberg, John J.

    2013-01-01

    Background: The Ramazzini Institute (RI) has completed nearly 400 cancer bioassays on > 200 compounds. The European Food Safety Authority (EFSA) and others have suggested that study design and protocol differences between the RI and other laboratories by may contribute to controversy regarding cancer hazard findings, principally findings on lymphoma/leukemia diagnoses. Objective: We aimed to evaluate RI study design, protocol differences, and accuracy of tumor diagnoses for their impact on carcinogenic hazard characterization. Methods: We analyzed the findings from a recent Pathology Working Group (PWG) review of RI procedures and tumor diagnoses, evaluated consistency of RI and other laboratory findings for chemicals identified by the RI as positive for lymphoma/leukemia, and examined evidence for a number of other issues raised regarding RI bioassays. The RI cancer bioassay design and protocols were evaluated in the context of relevant risk assessment guidance from international authorities. Discussion: Although the PWG identified close agreement with RI diagnoses for most tumor types, it did not find close agreement for lymphoma/leukemia of the respiratory tract or for neoplasms of the inner ear and cranium. Here we discuss a) the implications of the PWG findings, particularly lymphoma diagnostic issues; b) differences between RI studies and those from other laboratories that are relevant to evaluating RI cancer bioassays; and c) future work that may help resolve some concerns. Conclusions: We concluded that a) issues related to respiratory tract infections have complicated diagnoses at that site (i.e., lymphoma/leukemia), as well as for neoplasms of the inner ear and cranium, and b) there is consistency and value in RI studies for identification of other chemical-related neoplasia. Citation: Gift JS, Caldwell JC, Jinot J, Evans MV, Cote I, Vandenberg JJ. 2013. Scientific considerations for evaluating cancer bioassays conducted by the Ramazzini Institute

  3. Contact Bioassays with Phenoxybenzyl and Tetrafluorobenzyl Pyrethroids against Target-Site and Metabolic Resistant Mosquitoes

    PubMed Central

    Horstmann, Sebastian; Sonneck, Rainer

    2016-01-01

    Background Mosquito strains that exhibit increased tolerance to the chemical class of compounds with a sodium channel modulator mode of action (pyrethroids and pyrethrins) are typically described as “pyrethroid resistant”. Resistance to pyrethroids is an increasingly important challenge in the control of mosquito-borne diseases, such as malaria or dengue, because one of the main interventions (the distribution of large numbers of long-lasting insecticide-treated bed nets) currently relies entirely on long-lasting pyrethroids. Increasing tolerance of target insects against this class of insecticides lowers their impact in vector control. The current study suggests that the level of metabolic resistance depends on the structure of the molecule and that structurally different compounds may still be effective because detoxifying enzymes are unable to bind to these uncommon structures. Methods Treated surface contact bioassays were performed on susceptible Aedes aegypti, East African knockdown resistance (kdr) Anopheles gambiae (strain RSP-H) and metabolically resistant Anopheles funestus (strain FUMOZ-R) with different pyrethroids, such as cypermethrin, ß-cyfluthrin, deltamethrin, permethrin and transfluthrin (alone and in combination with the synergist piperonyl butoxide). The nonfluorinated form of transfluthrin was also assessed as a single agent and in combination with piperonyl butoxide. Results Although the dosages for pyrethroids containing a phenoxybenzyl moiety have exhibited differences in terms of effectiveness among the three tested mosquito species, the structurally different transfluthrin with a polyfluorobenzyl moiety remained active in mosquitoes with upregulated P450 levels. In trials with transfluthrin mixed with piperonyl butoxide, the added synergist exhibited no efficacy-enhancing effect. Conclusion The results of this study suggest that transfluthrin has the potential to control P450-mediated metabolically resistant mosquitoes because the

  4. SERS as a bioassay platform: fundamentals, design, and applications

    SciTech Connect

    Porter, M.; Lipert, R.; Siperko, L.; Wang, G.; Narayanan, R.

    2008-03-18

    Bioanalytical science is experiencing a period of unprecedented growth. Drivers behind this growth include the need to detect markers central to human and veterinary diagnostics at ever-lower levels and greater speeds. A set of parallel arguments applies to pathogens with respect to bioterrorism prevention and food and water safety. This tutorial review outlines our recent explorations on the use of surface enhanced Raman scattering (SERS) for detection of proteins, viruses, and microorganisms in heterogeneous immunoassays. It will detail the design and fabrication of the assay platform, including the capture substrate and nanoparticle-based labels. The latter, which is the cornerstone of our strategy, relies on the construction of gold nanoparticles modified with both an intrinsically strong Raman scatterer and an antibody. This labelling motif, referred to as extrinsic Raman labels (ERLs), takes advantage of the well-established signal enhancement of scatterers when coated on nanometre-sized gold particles, whereas the antibody imparts antigenic specificity. We will also examine the role of plasmon coupling between the ERLs and capture substrate, and challenges related to particle stability, nonspecific adsorption, and assay speed.

  5. Magnetic Bead Based Immunoassay for Autonomous Detection of Toxins

    SciTech Connect

    Kwon, Y; Hara, C A; Knize, M G; Hwang, M H; Venkatesteswaran, K S; Wheeler, E K; Bell, P M; Renzi, R F; Fruetel, J A; Bailey, C G

    2008-05-01

    As a step towards toward the development of a rapid, reliable analyzer for bioagents in the environment, we are developing an automated system for the simultaneous detection of a group of select agents and toxins. To detect toxins, we modified and automated an antibody-based approach previously developed for manual medical diagnostics that uses fluorescent eTag{trademark} reporter molecules and is suitable for highly multiplexed assays. Detection is based on two antibodies binding simultaneously to a single antigen, one of which is labeled with biotin while the other is conjugated to a fluorescent eTag{trademark} through a cleavable linkage. Aqueous samples are incubated with the mixture of antibodies along with streptavidin-coated magnetic beads coupled to a photo-activatable porphyrin complex. In the presence of antigen, a molecular complex is formed where the cleavable linkage is held in proximity to the photoactivable group. Upon excitation at 680 nm, free radicals are generated, which diffuse and cleave the linkage, releasing the eTags{trademark}. Released eTags{trademark} are analyzed using capillary gel electrophoresis with laser-induced fluorescence detection. Limits of detection for ovalbumin and botulinum toxoid individually were 4 ng/mL (or 80 pg) and 16 ng/mL (or 320 pg), respectively, using the manual assay. In addition, we demonstrated the use of pairs of antibodies from different sources in a single assay to decrease the rate of false positives. Automation of the assay was demonstrated on a flow-through format with higher LODs of 125 ng/mL (or 2.5 ng) each of a mixture of ovalbumin and botulinum toxoid. This versatile assay can be easily modified with the appropriate antibodies to detect a wide range of toxins and other proteins.

  6. Applicability of toxicity bioassays to ecological risk assessment in arid and semiarid ecosystems.

    SciTech Connect

    Markwiese, J. T.; Ryti, R. T.; Hooten, M. M.; Michael, D. I.; Hlohowskyj, I.; Environmental Assessment; Neptune and Company, Inc.

    2001-01-01

    Substantial tracts of land in the southwestern and western U.S. are undergoing or will require ERA. Toxicity bioassays employed in baseline ERAs are, for the most part. representative of mesic systems, and highly standardized test species (e.g., lettuce, earthworm) are generally not relevant to arid system toxicity testing. Conversely, relevant test species are often poorly characterized with regard to toxicant sensitivity and culture conditions. The applicability of toxicity bioassays to ecological risk assessment in arid and semiarid ecosystems was reviewed for bacteria and fungi, plants, terrestrial invertebrates, and terrestrial vertebrates. Bacteria and fungi are critical to soil processes, and understanding their ecology is important to understanding the ecological relevance of bioassays targeting either group. Terrestrial bacteria require a water film around soil particles to be active, while soil fungi can remain active in extremely dry soils. It is therefore expected that fungi will be of greater importance to arid and semiarid systems (Whitford 1989). If microbial processes are to be measured in soils of arid environments, it is recommended that bioassays target fungi. Regardless of the taxa studied, problems are associated with the standardization and interpretability of microbial tests, and regulatory acceptance may hinder widespread incorporation of microbial toxicity bioassays in arid system risk assessments. Plant toxicity bioassays are gaining recognition as sensitive indicators of soil conditions because they can provide a cost-effective and relatively rapid assessment of soil quality for both pre- and postremediation efforts. Although the choices of suitable plant species for assessing mesic system soils are numerous, the choices for arid system soils are limited. Guidance is provided for evaluating plant species with regard to their suitability for serving as representative arid system flora. Terrestrial invertebrates can survive and flourish in

  7. Interlaboratory comparison of in vitro bioassays for screening of endocrine active chemicals in recycled water.

    PubMed

    Mehinto, Alvine C; Jia, Ai; Snyder, Shane A; Jayasinghe, B Sumith; Denslow, Nancy D; Crago, Jordan; Schlenk, Daniel; Menzie, Christopher; Westerheide, Sandy D; Leusch, Frederic D L; Maruya, Keith A

    2015-10-15

    In vitro bioassays have shown promise as water quality monitoring tools. In this study, four commercially available in vitro bioassays (GeneBLAzer(®) androgen receptor (AR), estrogen receptor-alpha (ER), glucocorticoid receptor (GR) and progesterone receptor (PR) assays) were adapted to screen for endocrine active chemicals in samples from two recycled water plants. The standardized protocols were used in an interlaboratory comparison exercise to evaluate the reproducibility of in vitro bioassay results. Key performance criteria were successfully achieved, including low background response, standardized calibration parameters and high intra-laboratory precision. Only two datasets were excluded due to poor calibration performance. Good interlaboratory reproducibility was observed for GR bioassay, with 16-26% variability among the laboratories. ER and PR bioactivity was measured near the bioassay limit of detection and showed more variability (21-54%), although interlaboratory agreement remained comparable to that of conventional analytical methods. AR bioassay showed no activity for any of the samples analyzed. Our results indicate that ER, GR and PR, were capable of screening for different water quality, i.e., the highest bioactivity was observed in the plant influent, which also contained the highest concentrations of endocrine active chemicals measured by LC-MS/MS. After advanced treatment (e.g., reverse osmosis), bioactivity and target chemical concentrations were both below limits of detection. Comparison of bioassay and chemical equivalent concentrations revealed that targeted chemicals accounted for ≤5% of bioassay activity, suggesting that detection limits by LC-MS/MS for some chemicals were insufficient and/or other bioactive compounds were present in these samples. Our study demonstrated that in vitro bioassays responses were reproducible, and can provide information to complement conventional analytical methods for a more comprehensive water quality

  8. A reliable monitoring of the biocompatibility of an effluent along an oxidative pre-treatment by sequential bioassays and chemical analyses.

    PubMed

    Amat, A M; Arques, A; García-Ripoll, A; Santos-Juanes, L; Vicente, R; Oller, I; Maldonado, M I; Malato, S

    2009-02-01

    A new approach to assess biocompatibility of an effluent, based on combination of different bioassays and chemical analyses, has been tested using a mixture of four commercial pesticides treated by a solar photo-Fenton as target effluent. A very fast elimination of the pesticides occurred (all of them were below detection limit at t30W=36 min), but mineralisation was a more time-consuming process, due to the formation of organic intermediates and to the presence of solvents, as shown by GC-MS analysis. Measurements based on activated sludge indicated that detoxification was coincident with the removal of the active ingredients, while more sensitive Vibrio fischeri bacterium showed significant toxicity until the end of the experiment, although the effluent might be compatible with biological processes. Biodegradability of the solutions was enhanced by the photochemical process, to reach BOD5/COD ratios above 0.8. Longer time bioassays, such as the Zahn-Wellens' test, support the applicability of coupling photochemical with activated sludge-based biological processes to deal with these effluents. PMID:19070346

  9. [Investigation on pattern of quality control for Chinese materia medica based on famous-region drug and bioassay--the work reference].

    PubMed

    Yan, Dan; Xiao, Xiaohe

    2011-05-01

    Selection and standardization of the work reference are the technical issues to be faced with in the bioassay of Chinese materia medica. Taking the bioassay of Coptis chinensis. as an example, the manufacture process of the famous-region drugs extraction was explained from the aspects of original identification, routine examination, component analysis and bioassay. The common technologies were extracted, and the selection and standardization procedures of the work reference for the bioassay of Chinese materia medica were drawn up, so as to provide technical support for constructing a new mode and method of the quality control of Chinese materia medica based on the famous-region drugs and bioassay. PMID:21842660

  10. Toxicity bioassays for water from black-odor rivers in Wenzhou, China.

    PubMed

    DeFu, He; RuiRui, Chen; EnHui, Zhu; Na, Chen; Bo, Yang; HuaHong, Shi; MinSheng, Huang

    2015-02-01

    Following urbanization, a large number of urban rivers were contaminated and turned to black-odor rivers. The traditional approach for detecting water quality is based on chemical or physical analysis. However, biological toxicity of black-odor water has been less addressed. As two typical black-odor rivers, Jiushanwai River (JS) and Shanxia River (SX) are tributaries of Wen-Rui Tang River in Wenzhou (south of China). The eco-safety of the urban rivers was evaluated by bioassay for water toxicity in this study. Ten and 5 sampling sites were respectively set along JS and SX. Water samples were collected monthly from October 2010 to October 2011. The general physical and chemical parameters of river water were monitored. In order to investigate the ecotoxicological effects of black-odor water, the following bioassays were used: (1) Fish acute toxicity test (Danio rerio, comprehensive toxicity), (2) luminescent bacteria bioassay (Qinghaiensis vibrio, toxicity to bacteria), and (3) tropical claw embryo assay (Xenopus tropicalis, embryo toxicity). Biotoxicity of black-odor rivers water was demonstrated by D. rerio, Q. vibrio, and X. tropicalis embryos. Toxicological effects of black-odor water were respectively shown by mortality of zebrafish, and by the relative inhibitory light rate of luminescent bacteria. However, luminescent bacteria were more sensitive to inspect biotoxicity than zebrafish. In X. tropicalis embryos test, toxicological effects of black-odor water were mostly shown by embryos' survival rate and teratogenic rate. Bioassay results showed that toxicity of SX water was higher than that of JS water, especially in summer. Statistical analysis of luminescent bacteria toxicity test showed that biotoxicity of SX and JS was high in summer, but low in winter and spring. The seasonal changes of water toxicity of the black-odor river were positively correlative with changes of water temperature (p < 0.05), and related to pH and ammonium nitrogen of water

  11. Au@SiO2 core-shell structure involved with methotrexate: Fabrication, biodegradation process and bioassay explore.

    PubMed

    Huo, Xiaolei; Dai, Chaofan; Tian, Deying; Li, Shuping; Li, Xiaodong

    2015-12-30

    A new strategy is proposed to synthesize a kind of Au@SiO2 core-shell structure with methotrexate (MTX) loaded within it. Firstly, MTX molecules are attracted to the surface and vicinity of Au nanoparticles (NPs). Then the enriched MTX molecules on the surface of Au NPs have a good chance to be wrapped into the core-shell structure when SiO2 is uniformly deposited on the Au core. Secondly, the effect of Au amount and MTX content on the drug-loading capacity is emphatically studied and the result shows that core-shell structure plays a vital role in drug loading. In addition, the biodegradation process is also examined in phosphate buffer solution (PBS) at 37°C. The results show that the biodegradation of Au-MTX@SiO2 core-shell structure can be divided into two stages: the release of drug together with the fragmentation of core-shell structure and the subsequent dissolution of SiO2 layers. Lastly, in vitro bioassay tests give the evidence that obvious tumor inhibition can be achieved in presence of Au-MTX@SiO2 NPs even at low concentration and the efficacy can be greatly enhanced by the photothermal therapy on Au cores. PMID:26516099

  12. Magnetically labelled gold and epoxy bi-functional microcarriers for suspension based bioassay technologies.

    PubMed

    Vyas, Kunal N; Palfreyman, Justin J; Love, David M; Mitrelias, Thanos; Barnes, Crispin H W

    2012-12-21

    Microarrays and suspension-based assay technologies have attracted significant interest over the past decade with applications ranging from medical diagnostics to high throughput molecular biology. The throughput and sensitivity of a microarray will always be limited by the array density and slow reaction kinetics. Suspension (or bead) based technologies offer a conceptually different approach, improving detection by substituting a fixed plane of operation with many individually distinguishable microcarriers. In addition to all the features of a suspension based assay technology, our technology offers a rewritable label. This has the potential to be truly revolutionary by opening up the possibility of generating, on chip, extensive labelled molecular libraries. We unveil our latest SU-8 microcarrier design with embedded magnetic films that can be utilized for both magnetic and optical labelling. The novel design significantly simplifies fabrication and additionally incorporates a gold cap to provide a dual surface, bi-functional architecture. The microcarriers are fabricated using deep-ultraviolet lithography techniques and metallic thin film growth by evaporation. The bi-functional properties of the microcarriers will allow us to use each microcarrier as its own positive control thereby increasing the reliability of our technology. Here we present details of the design, fabrication, magnetic detection and functionalization of these microcarriers. PMID:23128508

  13. Inkjet-printed bioassays for direct reading with a multimode DVD/Blu-Ray optical drive.

    PubMed

    Li, Xiaochun; Shi, Maolin; Cui, Caie; Yu, Hua-Zhong

    2014-09-16

    Compact disc-based bioassays have been developed as novel point-of-care (POC) tools for various applications in chemical analysis and biomedical diagnosis. For the fabrication of assay discs, the surface patterning and sample introduction have been restricted to manual delivery that is unfavorable for on-demand high throughput medical screening. Herein, we have adapted a conventional inkjet printer to prepare bioassays on regular DVD-Rs and accomplished quantitative analysis with a multimode DVD/Blu-Ray optical drive in conjunction with free disc diagnostic software. The feasibility and accuracy of this method have been demonstrated by the quantitative analysis of inkjet-printed biotin-streptavidin binding assays on DVD, which serves as a trial system for other complex, medically relevant sandwich-format or competitive immunoassays. PMID:25144468

  14. Selection of a Suitable Disc Bioassay for the Screening of Anti-Tumor Molecules

    PubMed Central

    Trigui, Fatma; Pigeon, Pascal; Jalleli, Karim; Top, Siden; Aifa, Sami; El Arbi, Mehdi

    2013-01-01

    The crown gall induced in potato discs by Agrobacterium tumefaciens is becoming largely utilised in screening anti-tumor agents. The present work is showing that beet discs are more adequate for the anti-tumor screening test. In fact, maximal tumor induction was observed on beet discs (87.5%), followed by carrot discs (75%) and potato discs (68.5%). Beet discs present the most sensibility to crown gall disease with a fast expression of symptoms and more visible galls without any staining need. The beet discs bioassay was carried out by using some synthesized organometallics known for their antitumor activity in mammalian cells. We found significant crown gall inhibition (20.7% to 40.55%) of the tested compounds. Overall results supported that beet bioassay might be a potential prescreen system of anti-tumor molecules in mammalian cells. PMID:24711759

  15. Labor-effective manipulation of marine and beetle luciferases for bioassays.

    PubMed

    Kim, Sung Bae

    2012-06-01

    Engineering of luciferases with designed properties and functionalities collects great interest in bioassays. However, such an engineering including mutagenesis accompanies great consumption of time-and-labor. Here, I review an empirical approach to efficiently manipulate marine and beetle luciferases for bioassays, where a putative active site of luciferases is initially assigned with an in silico analysis, prior to the practical engineering, e.g. a hydrophilicity search reveals a characteristic hydrophilic region of luciferases as an engineering target. Amino acids in the hydrophilic region are recommended for a mutagenesis target to generate superluminescent variants of marine luciferases with prolonged bioluminescence. Empirical data suggest that a consecutive fragmentation to the assigned hydrophilic site greatly reduces time-and-labors on construction of single-chain probes. This review summarizes how to relieve the efforts for fabricating single-chain probes and potent variants of luciferases with excellent optical properties. PMID:22514115

  16. Regional differences in stratum corneum reactivity to surfactants. Quantitative assessment using the corneosurfametry bioassay.

    PubMed

    Henry, F; Goffin, V; Maibach, H I; Piérard, G E

    1997-12-01

    The skin does not react similarly to the presence of xenobiotics over all anatomic sites. Distinct regional differences have been described for irritancy and percutaneous absorption. The present study assesses the regional variation of stratum corneum reactivity to surfactants using the corneosurfametry bioassay. Stratum corneum was harvested from 6 body sites in 20 young adults. Corneosurfametry was performed using water, 1% SLS and a 5% soap solution. Data show that the best variable to assess regional variability in irritancy is the overall difference in corneosurfametry (ODC), comparing the effect of a given surfactant with water. The dorsal hand and volar forearm were the least reactive, the neck, forehead, back and dorsal foot the most reactive, sites. It is concluded that the corneosurfametry bioassay, through the ODC variable, is a practically noninvasive tool for the evaluation of regional variation in irritancy at the level of the stratum corneum. PMID:9455629

  17. Compost maturity assessment using physicochemical, solid-state spectroscopy, and plant bioassay analysis.

    PubMed

    Kumar, D Senthil; Kumar, P Satheesh; Rajendran, N M; Anbuganapathi, G

    2013-11-27

    The vermicompost produced from flower waste inoculated with biofertilizers was subjected to compost maturity test: (i) physicochemical method (pH, OC, TN, C:N); (ii) solid state spectroscopic analysis (FTIR and (13)C CPMAS NMR); and (iii) plant bioassay (germination index). The pH of vermicompost was decreased toward neutral, C:N ratio < 20; reduced organic carbon with increased nitrogen indicates the compost attains maturity. The final vermicomposts result shows reduction of complex organic materials into simple minerals which indicates the maturity of the experimental vermicompost product than the control. The increased aliphatic portion incorporated with flower residues may be due to the synthesis of alkyl, O-alkyl, and COO groups by the microbes present in the gut of earthworm. Plant bioassays are considered the most conventional assessment of compost maturity analysis, and subsequently, it shows the effect of vermicompost maturity on the germination index of Vigna mungo . PMID:24191667

  18. Characterizing the genotoxicity of hazardous industrial wastes and effluents using short-term bioassays

    SciTech Connect

    Houk, V.S.; DeMarini, D.M.

    1989-01-01

    This paper demonstrates that short-term bioassays can reliably and expeditiously measure the genotoxic potential of hazardous industrial wastes and effluents. Petrochemical wastes have been studied in detail, especially discharges from chemical manufacturing plants and textile and dye effluents. However, there is little information on effluents from pesticide manufacturers. The most extensive evaluations have been conducted on effluents from pulp and paper mills. These studies have shown which pulping plants generate the most genotoxic effluents, which process wastes are most hazardous, have isolated and identified the compounds responsible for the genotoxic activity, have described the environmental fate of these compounds, have evaluated the types of genetic damage likely to occur upon exposure to the effluents, and have identified several treatment methods that effectively reduce the genotoxicity of the effluents. The coupling of bioassays for biological analysis with chemical evaluation provides the most powerful approach to assessing the overall health effects of complex industrial wastes and effluents.

  19. Lung tumors in strain A mice as a bioassay for carcinogenicity of environmental chemicals

    SciTech Connect

    Stoner, G.D. )

    1991-03-01

    This report describes the protocol for the strain A mouse lung tumor bioassay and summarizes results on selected chemicals that have been tested for carcinogenicity in the assay. The assay is of 6 months duration and can distinguish 2-fold differences in carcinogenic potential of compounds from several chemical classes. Specifically, the assay is sensitive to polycyclic hydrocarbons, nitrosamines and nitrosoureas, carbamates, aflatoxin, certain metals, hydrazines, and others, but is relatively insensitive to aromatic amines, aliphatic halides, and other compounds that are carcinogenic in the rodent liver and/or bladder. Recommendations are made for future studies on the: (1) distribution and metabolism of chemicals in strain A mouse lung tissue and in specific lung cell types; (2) ability of the lung tumor bioassay to detect inhibitors and promoters of carcinogenesis; and (3) use of the assay for testing mixtures of chemicals for carcinogenic activity.

  20. Evaluation of mouse bioassay results in an inter-laboratory comparison for paralytic shellfish poisoning toxins

    NASA Astrophysics Data System (ADS)

    Cao, Jijuan; Zheng, Jiang; Yu, Bing; Wang, Qiuyan; Xu, Junyi; Li, Aifeng

    2011-07-01

    An inter-laboratory comparison of the AOAC mouse bioassay for paralytic shellfish poisoning (PSP) toxicity in shellfish was carried out among 25 Chinese laboratories to examine the overall performance for PSP testing in China, and to analyze the main factors affecting the performance of this method. The toxic scallop Patinopecten yessoensis collected from coast of Bohai Sea, China, was used as a test sample in the comparison study. The results were reported and evaluated using robust statistical methods. The z scores showed that 80%, 8%, and 12% of laboratories reported satisfactory results, unsatisfactory results, and questionable results, respectively. This evaluation demonstrates that the PSP mouse bioassay is an appropriate method for screening and testing PSP toxicity in shellfish. However, it was found that the experience and skill of technicians, as well as the body weight and health status of mice being used significantly affected the accuracy of the method.

  1. Sensitive new in vitro bioassay for melanocyte-stimulating activity using the skin of Anolis carolinensis.

    PubMed

    Carter, R J; Shuster, S

    1978-10-01

    An accurate, highly reproducible and sensitive bioassay for melanocyte-stimulating hormone (MSH) using the skin of Anolis carolinensis in vitro is described. The time taken for green Anolis skin fragments to change to a specific, visually assessed, green-brown color is dose-related, and this forms the basis of the new assay. The method is simple to perform, and 1 person may assay 20 samples in a day using the dorsal skin from a single adult lizard. The mean dose-response ranges between 48 X 10(-12) and 375 X 12(-12) M (38 to 625 pg/ml). Using the assay, alpha-MSH, beta-MSH, ACTH (4-10), and ACTH (1-10) were equipotent on a molar basis. For repeated bioassay of rat pituitary extracts, the dose-response curves were highly significant, and only 1 of the 9 pituitary dose-response curves deviated significantly from the slope of the standard alpha-MSH curve. The index of precision, lambda, for the 9 pituitary bioassays ranged between 0.037 and 0.081, while the mean 95% fiducial limits were -6.6 and 7.1% on either side of the estimated potency. The new rate method is compared with an earlier quantal method which also uses the isolated skin of Anolis carolinensis. The quantal method does not have dose-response characteristics and is therefore less accurate and reproducible than the new method; the coefficient of variation for repeated bioassay of the same pituitary extracts ranged from between 12 to 20% for the quantal method and between 2.9 to 5.7% for the new rate method. PMID:212484

  2. Carcinogenic risk of copper gluconate evaluated by a rat medium-term liver carcinogenicity bioassay protocol.

    PubMed

    Abe, Masayoshi; Usuda, Koji; Hayashi, Seigo; Ogawa, Izumi; Furukawa, Satoshi; Igarashi, Maki; Nakae, Dai

    2008-08-01

    Carcinogenic risk and molecular mechanisms underlying the liver tumor-promoting activity of copper gluconate, an additive of functional foods, were investigated using a rat medium-term liver carcinogenicity bioassay protocol (Ito test) and a 2-week short-term administration experiment. In the medium-term liver bioassay, Fischer 344 male rats were given a single i.p. injection of N-nitrosodiethylamine at a dose of 200 mg/kg b.w. as a carcinogenic initiator. Starting 2 weeks thereafter, rats received 0, 10, 300 or 6,000 ppm of copper gluconate in diet for 6 weeks. All rats underwent 2/3 partial hepatectomy at the end of week 3, and all surviving rats were killed at the end of week 8. In the short-term experiment, rats were given 0, 10, 300 or 6,000 ppm of copper gluconate for 2 weeks. Numbers of glutathione S-transferase placental form (GST-P) positive lesions, single GST-P-positive hepatocytes and 8-oxoguanine-positive hepatocytes, and levels of cell proliferation and apoptosis in the liver were significantly increased by 6,000 ppm of copper gluconate in the medium-term liver bioassay. Furthermore, hepatic mRNA expression of genes relating to the metal metabolism, inflammation and apoptosis were elevated by 6,000 ppm of copper gluconate both in the medium-term liver bioassay and the short-term experiments. These results indicate that copper gluconate possesses carcinogenic risk toward the liver at the high dose level, and that oxidative stress and inflammatory and pro-apoptotic signaling statuses may participate in its underlying mechanisms. PMID:18350280

  3. The OECD program to validate the rat uterotrophic bioassay. Phase 2: dose-response studies.

    PubMed Central

    Kanno, Jun; Onyon, Lesley; Peddada, Shyamal; Ashby, John; Jacob, Elard; Owens, William

    2003-01-01

    The Organisation for Economic Co-operation and Development has completed phase 2 of an international validation program for the rodent uterotrophic bioassay. The purpose of the validation program was to demonstrate the performance of two versions of the uterotrophic bioassay, the immature female rat and the adult ovariectomized rat, in four standardized protocols. This article reports the dose-response studies of the validation program; the coded single-dose studies are reported in an accompanying paper. The dose-response study design used five selected weak estrogen agonists, bisphenol A, genistein, methoxychlor, nonylphenol, and o,p -DDT. These weak agonists were administered in a prescribed series of doses to measure the performance and reproducibility of the protocols among the participating laboratories. All protocols successfully detected increases in uterine weights when the weak agonists were administered. Within each protocol, there was good agreement and reproducibility of the dose response among laboratories with each substance. Substance-specific variations were observed in the influence of the route of administration on the uterine response, the potency as related to the dose producing the first statistically significant increase in uterine weights, and the maximum increase in uterine weight. Substantive performance differences were not observed between the uterotrophic bioassay versions or among the standardized protocols, and these were judged to be qualitatively equivalent. It is noteworthy that these results were reproducible under a variety of different experimental conditions (e.g., animal strain, diet, housing, bedding, vehicle, animal age), indicating that the bioassay's performance as a screen is robust. In conclusion, both the intact, immature, and adult OVX versions, and all protocols appear to be reproducible and transferable across laboratories and are able to detect weak estrogen agonists. PMID:12948896

  4. Zebrafish (Danio rerio) bioassay for visceral toxicosis of catfish and botulinum neurotoxin serotype E.

    PubMed

    Chatla, Kamalakar; Gaunt, Patricia; Petrie-Hanson, Lora; Hohn, Claudia; Ford, Lorelei; Hanson, Larry

    2014-03-01

    Visceral toxicosis of catfish (VTC), a sporadic disease of cultured channel catfish (Ictalurus punctatus) often with high mortality, is caused by botulinum neurotoxin serotype E (BoNT/E). Presumptive diagnosis of VTC is based on characteristic clinical signs and lesions, and the production of these signs and mortality after sera from affected fish is administered to sentinel catfish. The diagnosis is confirmed if the toxicity is neutralized with BoNT/E antitoxin. Because small catfish are often unavailable, the utility of adult zebrafish (Danio rerio) was evaluated in BoNT/E and VTC bioassays. Channel catfish and zebrafish susceptibilities were compared using trypsin-activated BoNT/E in a 96-hr trial by intracoelomically administering 0, 1.87, 3.7, 7.5, 15, or 30 pg of toxin per gram of body weight (g-bw) of fish. All of the zebrafish died at the 7.5 pg/g-bw and higher, while the catfish died at the 15 pg/g-bw dose and higher. To test the bioassay, sera from VTC-affected fish or control sera were intracoelomically injected at a dose of 10 µl per zebrafish and 20 µl/g-bw for channel catfish. At 96 hr post-injection, 78% of the zebrafish and 50% of the catfish receiving VTC sera died, while no control fish died. When the VTC sera were preincubated with BoNT/E antitoxin, they became nontoxic to zebrafish. Histology of zebrafish injected with either VTC serum or BoNT/E demonstrated renal necrosis. Normal catfish serum was toxic to larval zebrafish in immersion exposures, abrogating their utility in VTC bioassays. The results demonstrate bioassays using adult zebrafish for detecting BoNT/E and VTC are sensitive and practical. PMID:24518279

  5. DSSTOX NATIONAL TOXICOLOGY PROGRAM BIOASSAY ON-LINE DATABASE STRUCTURE-INDEX LOCATOR FILE: SDF FILE AND DOCUMENTATION

    EPA Science Inventory

    NTPBSI: National Toxicology Program Bioassay On-line Database Structure-Index Locator File. Database contains the results collected on approxiately 300 toxicity studies from shorter duration test and from genetic toxicity studies, both in vitro and in vivo tests.

  6. Fluorescence-based bioassays for the detection and evaluation of food materials.

    PubMed

    Nishi, Kentaro; Isobe, Shin-Ichiro; Zhu, Yun; Kiyama, Ryoiti

    2015-01-01

    We summarize here the recent progress in fluorescence-based bioassays for the detection and evaluation of food materials by focusing on fluorescent dyes used in bioassays and applications of these assays for food safety, quality and efficacy. Fluorescent dyes have been used in various bioassays, such as biosensing, cell assay, energy transfer-based assay, probing, protein/immunological assay and microarray/biochip assay. Among the arrays used in microarray/biochip assay, fluorescence-based microarrays/biochips, such as antibody/protein microarrays, bead/suspension arrays, capillary/sensor arrays, DNA microarrays/polymerase chain reaction (PCR)-based arrays, glycan/lectin arrays, immunoassay/enzyme-linked immunosorbent assay (ELISA)-based arrays, microfluidic chips and tissue arrays, have been developed and used for the assessment of allergy/poisoning/toxicity, contamination and efficacy/mechanism, and quality control/safety. DNA microarray assays have been used widely for food safety and quality as well as searches for active components. DNA microarray-based gene expression profiling may be useful for such purposes due to its advantages in the evaluation of pathway-based intracellular signaling in response to food materials. PMID:26473869

  7. Toxicity assessment through multiple endpoint bioassays in soils posing environmental risk according to regulatory screening values.

    PubMed

    Rodriguez-Ruiz, A; Asensio, V; Zaldibar, B; Soto, M; Marigómez, I

    2014-01-01

    Toxicity profiles of two soils (a brownfield in Legazpi and an abandoned iron mine in Zugaztieta; Basque Country) contaminated with several metals (As, Zn, Pb and Cu in Legazpi; Zn, Pb, Cd and Cu in Zugaztieta) and petroleum hydrocarbons (in Legazpi) were determined using a multi-endpoint bioassay approach. Investigated soils exceeded screening values (SVs) of regulatory policies in force (Basque Country; Europe). Acute and chronic toxicity bioassays were conducted with a selected set of test species (Vibrio fischeri, Dictyostelium discoideum, Lactuca sativa, Raphanus sativus and Eisenia fetida) in combination with chemical analysis of soils and elutriates, as well as with bioaccumulation studies in earthworms. The sensitivity of the test species and the toxicity endpoints varied depending on the soil. It was concluded that whilst Zugaztieta soil showed very little or no toxicity, Legazpi soil was toxic according to almost all the toxicity tests (solid phase Microtox, D. discoideum inhibition of fruiting body formation and developmental cycle solid phase assays, lettuce seed germination and root elongation test, earthworm acute toxicity and reproduction tests, D. discoideum cell viability and replication elutriate assays). Thus, albeit both soils had similar SVs, their ecotoxicological risk, and therefore the need for intervening, was different for each soil as unveiled after toxicity profiling based on multiple endpoint bioassays. Such a toxicity profiling approach is suitable to be applied for scenario-targeted soil risk assessment in those cases where applicable national/regional soil legislation based on SVs demands further toxicity assessment. PMID:24819436

  8. Bioassay directed identification of natural aryl hydrocarbon-receptor agonists in marmalade.

    PubMed

    van Ede, Karin; Li, An; Antunes-Fernandes, Elsa; Mulder, Patrick; Peijnenburg, Ad; Hoogenboom, Ron

    2008-06-01

    Citrus fruit and citrus fruit products, like grapefruit, lemon and marmalade were shown to contain aryl hydrocarbon receptor (AhR) agonists, as detected with the DR CALUX bioassay. This is of interest regarding the role of the Ah-receptor pathway in the adverse effects of dioxins, PCBs and other aromatic hydrocarbons. So far it is unclear which compounds in citrus fruit are responsible for the AhR-mediated activity and whether regular exposure to these compounds can cause effects comparable to, e.g. dioxins. The present study aimed at developing a method for identifying unknown Ah-receptor agonists in citrus products based on bioassay directed analysis, using marmalade as a first target. Following extraction with hexane and purification on an aluminium oxide-column, the extract was fractionated by HPLC using a C-18 semi-preparative column. Fractions were extracted, solvent-exchanged into dimethylsulfoxide and subsequently tested with DR CALUX. Extracts were shown to contain primarily coumarins, furocoumarins (FCs) and polymethoxyflavones (PMFs). Identification of fractions most active in the bioassay via LC/MS revealed that bergapten (an FC) is the most important Ah-receptor agonist in marmalade. The approach and method developed resulted in the successful identification of the bioactive component. However, potential pitfalls of the procedure will be discussed. PMID:18486664

  9. Bioassay assessment of metarhizium anisopliae (metchnikoff) sorokin (deuteromycota: hyphomycetes) against Oncometopia facialis (signoret) (hemiptera: cicadellidae)

    PubMed Central

    Pria Júnior, Wolney Dalla; Lacava, Paulo Teixeira; Messias, Claudio Luiz; Azevedo, João Lúcio; Lacava, Pedro Magalhães

    2008-01-01

    Citrus Variegated Chlorosis (CVC) is an economically important, destructive disease in Brazil and is caused by Xylella fastidiosa and transmitted by sharpshooter insects. In this study, the efficacy of the fungus Metarhizium anisopliae in controlling the sharpshooter Oncometopia facialis was studied by bioassay conditions. In the bioassay, insects were sprayed with a suspension containing 5 X 107 conidia mL-1. Adults captured in the field were treated in groups of 10 in a total of 11 replications per treatment. Significant differences between the natural mortality and the mortality of insects treated with the fungus were observed 6 days after inoculations (P<0.05). These significant differences increased until 10 days after treatment. The fungus caused 87.1% mortality, with the LT50 varying from 5 to 6 days. The LC50 was 1.2 X 106 conidia mL-1, varying from 7.7 X 105 to 2 X 106 conidia mL-1. The results showed that the sharpshooter O. facialis was susceptible to the entomopathogenic action of M. anisopliae in controlled condition during bioassay. PMID:24031192

  10. Bioassays as one of the Green Chemistry tools for assessing environmental quality: A review.

    PubMed

    Wieczerzak, M; Namieśnik, J; Kudłak, B

    2016-09-01

    For centuries, mankind has contributed to irreversible environmental changes, but due to the modern science of recent decades, scientists are able to assess the scale of this impact. The introduction of laws and standards to ensure environmental cleanliness requires comprehensive environmental monitoring, which should also meet the requirements of Green Chemistry. The broad spectrum of Green Chemistry principle applications should also include all of the techniques and methods of pollutant analysis and environmental monitoring. The classical methods of chemical analyses do not always match the twelve principles of Green Chemistry, and they are often expensive and employ toxic and environmentally unfriendly solvents in large quantities. These solvents can generate hazardous and toxic waste while consuming large volumes of resources. Therefore, there is a need to develop reliable techniques that would not only meet the requirements of Green Analytical Chemistry, but they could also complement and sometimes provide an alternative to conventional classical analytical methods. These alternatives may be found in bioassays. Commercially available certified bioassays often come in the form of ready-to-use toxkits, and they are easy to use and relatively inexpensive in comparison with certain conventional analytical methods. The aim of this study is to provide evidence that bioassays can be a complementary alternative to classical methods of analysis and can fulfil Green Analytical Chemistry criteria. The test organisms discussed in this work include single-celled organisms, such as cell lines, fungi (yeast), and bacteria, and multicellular organisms, such as invertebrate and vertebrate animals and plants. PMID:27472199

  11. Bivalve embryo bioassay to assess the potential toxicity of dredged material before dumping

    SciTech Connect

    Quiniou, F.

    1995-12-31

    Dredged harbor sediments frequently contain a wide spectrum of contaminants in addition to a significant percentage of organic matter. Also, dredging and dumping activities into sea water, of these highly contaminated soil may induce a harmful effect on the environment. In France, in accordance with Oslo convention guidelines, a working group on dredging activities and environment (GEODE) created since 1991 decided to set up a pilot research program to assess the intrinsic toxicity of four harbor sludges. Intrinsic toxicity of harbor muds were tested by solid phase (whole sediment) and aqueous extract bioassays (sea water elutriates) using the sublethal toxicity test bivalve embryo bioassay (Crassostrea gigas). Elutriates enable them to detect the toxicity of contaminants which may be released in the soluble form into the water column during dredging operations. While, whole sediment integrate the synergistic effects of all the contaminants (hydrophilic and hydrophobic) including pore water. Bioassays results, correlated to chemical analysis, are compared to contaminant levels determined by French working group GEODE and Canadian sediment quality criteria.

  12. Fluorescence-Based Bioassays for the Detection and Evaluation of Food Materials

    PubMed Central

    Nishi, Kentaro; Isobe, Shin-Ichiro; Zhu, Yun; Kiyama, Ryoiti

    2015-01-01

    We summarize here the recent progress in fluorescence-based bioassays for the detection and evaluation of food materials by focusing on fluorescent dyes used in bioassays and applications of these assays for food safety, quality and efficacy. Fluorescent dyes have been used in various bioassays, such as biosensing, cell assay, energy transfer-based assay, probing, protein/immunological assay and microarray/biochip assay. Among the arrays used in microarray/biochip assay, fluorescence-based microarrays/biochips, such as antibody/protein microarrays, bead/suspension arrays, capillary/sensor arrays, DNA microarrays/polymerase chain reaction (PCR)-based arrays, glycan/lectin arrays, immunoassay/enzyme-linked immunosorbent assay (ELISA)-based arrays, microfluidic chips and tissue arrays, have been developed and used for the assessment of allergy/poisoning/toxicity, contamination and efficacy/mechanism, and quality control/safety. DNA microarray assays have been used widely for food safety and quality as well as searches for active components. DNA microarray-based gene expression profiling may be useful for such purposes due to its advantages in the evaluation of pathway-based intracellular signaling in response to food materials. PMID:26473869

  13. Short-term bioassays of fractionated emission samples from wood combustion

    SciTech Connect

    Alfheim, I.; Becher, G.; Hongslo, J.K.; Lazaridis, G.; Loefroth, G.; Ramdahl, T.; Rivedal, E.; Salomaa, S.; Sanner, T.; Sorsa, M.

    1984-01-01

    Extracts of an emission sample from wood burning, consisting of particles and volatiles, have been fractionated on an HPLC silica gel column into five fractions of increasing polarity. Nonfractionated samples and the individual fractions have been tested in three different short-term bioassays: the Ames Salmonella assay, the sister chromatid exchange (SCE) induction-test in Chinese hamster ovary cells (CHO), and the cell transformation test on Syrian hamster embryo (SHE) cells. Most of the total activity was found in the volatile part of the sample with all three bioassays, whereas the particle extract had the highest activity per unit mass extracted. The second most polar fraction contained most of the mass and was also highly active in all assays. The most polar fraction was very potent in the Salmonella assay, but showed only a weak response in the eukaryotic bioassays. Storage of the samples for several months at 0 degrees C revealed that the bacterial mutagens present in the most polar fraction were labile; the mutagenicity was almost totally lost after 1 year's storage.

  14. Glycomics Approaches for the Bioassay and Structural Analysis of Heparin/Heparan Sulphates

    PubMed Central

    Puvirajesinghe, Tania M.; Turnbull, Jeremy E.

    2012-01-01

    The glycosaminoglycan heparan sulphate (HS) has a heterogeneous structure; evidence shows that specific structures may be responsible for specific functions in biological processes such as blood coagulation and regulation of growth factor signalling. This review summarises the different experimental tools and methods developed to provide more rapid methods for studying the structure and functions of HS. Rapid and sensitive methods for the facile purification of HS, from tissue and cell sources are reviewed. Data sets for the structural analysis are often complex and include multiple sample sets, therefore different software and tools have been developed for the analysis of different HS data sets. These can be readily applied to chromatographic data sets for the simplification of data (e.g., charge separation using strong anion exchange chromatography and from size separation using gel filtration techniques. Finally, following the sequencing of the human genome, research has rapidly advanced with the introduction of high throughput technologies to carry out simultaneous analyses of many samples. Microarrays to study macromolecular interactions (including glycan arrays) have paved the way for bioassay technologies which utilize cell arrays to study the effects of multiple macromolecules on cells. Glycan bioassay technologies are described in which immobilisation techniques for saccharides are exploited to develop a platform to probe cell responses such as signalling pathway activation. This review aims at reviewing available techniques and tools for the purification, analysis and bioassay of HS saccharides in biological systems using “glycomics” approaches. PMID:24957775

  15. Bioassays as a tool for evaluating advanced oxidation processes in water and wastewater treatment.

    PubMed

    Rizzo, Luigi

    2011-10-01

    Advanced oxidation processes (AOPs) have been widely used in water and wastewater treatment for the removal of organic and inorganic contaminants as well as to improve biodegradability of industrial wastewater. Unfortunately, the partial oxidation of organic contaminants may result in the formation of intermediates more toxic than parent compounds. In order to avoid this drawback, AOPs are expected to be carefully operated and monitored, and toxicity tests have been used to evaluate whether effluent detoxification takes place. In the present work, the effect of AOPs on the toxicity of aqueous solutions of different classes of contaminants as well as actual aqueous matrices are critically reviewed. The dualism toxicity-biodegradability when AOPs are used as pre-treatment step to improve industrial wastewater biodegradability is also discussed. The main conclusions/remarks include the followings: (i) bioassays are a really useful tool to evaluate the dangerousness of AOPs as well as to set up the proper operative conditions, (ii) target organisms for bioassays should be chosen according to the final use of the treated water matrix, (iii) acute toxicity tests may be not suitable to evaluate toxicity in the presence of low/realistic concentrations of target contaminants, so studies on chronic effects should be further developed, (iv) some toxicity tests may be not useful to evaluate biodegradability potential, in this case more suitable tests should be applied (e.g., activated sludge bioassays, respirometry). PMID:21722938

  16. [Anti-platelet aggregation bioassay based quality control for XST capsules].

    PubMed

    Han, Bing; Mao, Xin; Han, Shu-xian; Chen, Ying; Xiang, Yan-hua; Ge, Yi-meng; Liao, Fu-long; You, Yun

    2015-12-01

    A in vitro platelet aggregation bioassay was developed for the quality control of XST capsules. The in vitro anti-platelet aggregation effect in rats was observed to detect the bioactivity of XST capsules. Panax notoginseng saponins and Xuesaitong lyophilizedpowder for injection were taken as standard control substances to determine the potency. According to the results, XST capsules showeda significant inhibitory effect on thrombin-induced platelet aggregation in a dose-dependent manner. The in vitro anti-platelet activity oflyophilized powder for injection was stabler than that of Panax notoginseng saponins, and so suitable to serve as a standard control substance. The biological potency of XST capsules compared with standard control substance was detected by using parallel line assay. According to the results, the established bioassay method had a good repeatability (RSD 2.92%). The sample test results could pass thereliability test(linear deviation P > 0.05, parallel deviation P > 0.05). This bioassay method could be used as one of the complementary quality control methods for XST capsules. PMID:27141669

  17. Identifying chemical carcinogens and assessing potential risk in short-term bioassays using transgenic mouse models.

    PubMed Central

    Tennant, R W; French, J E; Spalding, J W

    1995-01-01

    Cancer is a worldwide public health concern. Identifying carcinogens and limiting their exposure is one approach to the problem of reducing risk. Currently, epidemiology and rodent bioassays are the means by which putative human carcinogens are identified. Both methods have intrinsic limitations: they are slow and expensive processes with many uncertainties. The development of methods to modify specific genes in the mammalian genome has provided promising new tools for identifying carcinogens and characterizing risk. Transgenic mice may provide advantages in shortening the time required for bioassays and improving the accuracy of carcinogen identification; transgenic mice might now be included in the testing armamentarium without abandoning the two-year bioassay, the current standard. We show that mutagenic carcinogens can be identified with increased sensitivity and specificity using hemizygous p53 mice in which one allele of the p53 gene has been inactivated. Furthermore, the TG.AC transgenic model, carrying a v-Ha-ras construct, has developed papillomas and malignant tumors in response to a number of mutagenic and nonmutagenic carcinogens and tumor promoters, but not to noncarcinogens. We present a decision-tree approach that permits, at modest extra cost, the testing of more chemicals with improved ability to extrapolate from rodents to humans. Images Figure 1. Figure 2. Figure 3. PMID:8529591

  18. Experimental and Computational Characterization of Biological Liquid Crystals: A Review of Single-Molecule Bioassays

    PubMed Central

    Eom, Kilho; Yang, Jaemoon; Park, Jinsung; Yoon, Gwonchan; Soo Sohn, Young; Park, Shinsuk; Yoon, Dae Sung; Na, Sungsoo; Kwon, Taeyun

    2009-01-01

    Quantitative understanding of the mechanical behavior of biological liquid crystals such as proteins is essential for gaining insight into their biological functions, since some proteins perform notable mechanical functions. Recently, single-molecule experiments have allowed not only the quantitative characterization of the mechanical behavior of proteins such as protein unfolding mechanics, but also the exploration of the free energy landscape for protein folding. In this work, we have reviewed the current state-of-art in single-molecule bioassays that enable quantitative studies on protein unfolding mechanics and/or various molecular interactions. Specifically, single-molecule pulling experiments based on atomic force microscopy (AFM) have been overviewed. In addition, the computational simulations on single-molecule pulling experiments have been reviewed. We have also reviewed the AFM cantilever-based bioassay that provides insight into various molecular interactions. Our review highlights the AFM-based single-molecule bioassay for quantitative characterization of biological liquid crystals such as proteins. PMID:19865530

  19. An evaluation of the effectiveness of utilizing bioassays in the assessment of contaminated sites

    SciTech Connect

    Mroz, R.; Carter, J.; Tay, K.L.; Doe, K.

    1995-12-31

    The purpose of this study was to evaluate the battery of biological tests recommended by Environment Canada in the document ``A Review of Whole Organism Bioassays for Assessing the Quality of Soil, Freshwater Sediment and Freshwater in Canada`` for the assessment of contaminated sites. Soil and sediment samples were collected from three contaminated sites in the Atlantic Region and subjected to biological and chemical tests. Four bioassays were conducted on the soil samples: lettuce (Lactuca sativa) seedling emergence, algal (Selenastrum capricornutum) population growth inhibition, earthworm (Eisenia andrel) survival and inhibition of light output in Microtox (Vibrio fischeri). Soil samples collected from Makinsons, Newfoundland had elevated levels of PCBs, total petroleum hydrocarbons (TPH) and heavy metals and showed some toxicity in the algal population growth inhibition test. Samples from the Weldon, New Brunswick site were high in TPH and were marginally toxic to Microtox and lettuce seedlings. The earthworm survival test did not appear sensitive to any of the contaminated soil samples. Freshwater sediment samples, collected from Five Island Lake, Nova Scotia had elevated PCB and heavy metal concentrations. These samples underwent four biological tests: midge (Chironomus tentans) survival, amphipod (Hyalella azteca) survival, algal population growth inhibition and Microtox. At 100% concentration, the sediment was toxic to the first three species, with toxicities ranging from marginal to high. For all samples, the bioassay results were compared to chemical analyses and, in most cases, there was a positive correlation between contaminant concentrations and toxicity.

  20. Chronic bioassays of rainbow trout fry with compounds representative of contaminants in Great Lakes fish

    USGS Publications Warehouse

    Passino-Reader, Dora R.; Berlin, William H.; Hickey, James P.

    1995-01-01

    To evaluate the hazard of organic compounds detected in Great Lakes fish by gas chromatography/mass spectrometry, we tested compounds representative of heterocyclic nitrogen compounds, polycyclic aromatic hydrocarbons, and cyclic alkanes and alkenes. Sixty-day bioassays on the effects of nicotine, phenanthrene, pinane, and pinene on the behavior, growth, and survival of rainbow trout fry, Oncorhynchus mykiss, were conducted in a large, constant-flow, temperature-controlled water system. The following 60-day LCSO's were determined (mg/L): nicotine 5.0, phenanthrene 0.2, pinane 0.8, and pinene 1.2. Values of lowest observed effects level (LOEL) and no observed effects level (NOEL) showed that growth was generally as sensitive an endpoint as behavior and was more sensitive than time of swim-up. The 60-day LC50 values for rainbow trout were compared with earlier acute bioassays with Daphnia pulexand rainbow trout and chronic bioassays with D.pulex conducted at the Great Lakes Science Center. Rainbow trout fry were less sensitive than daphnids in all tests, indicating that toxicity tests with daphnids should be protective of salmonid fry for these types of compounds. The results for representative compounds indicate that these classes of compounds should be included in aquatic risk assessments at sites in the Great Lakes.

  1. Enhancement of the Chemiluminescence Response of Enzymatic Reactions by Plasmonic Surfaces for Biosensing Applications

    PubMed Central

    Abel, Biebele; Odukoya, Babatunde; Mohammed, Muzaffer; Aslan, Kadir

    2015-01-01

    We report the enhancement of chemiluminescence response of horseradish peroxidase (HRP) in bioassays by plasmonic surfaces, which are comprised of (i) silver island films (SIFs) and (ii) metal thin films (silver, gold, copper, and nickel, 1 nm thick) deposited onto glass slides. A model bioassay, based on the interactions of avidin-modified HRP with a monolayer of biotinylated poly(ethylene-glycol)-amine, was employed to evaluate the ability of plasmonic surfaces to enhance chemiluminescence response of HRP. Chemiluminescence response of HRP in model bioassays were increased up to ~3.7-fold as compared to the control samples (i.e. glass slides without plasmonic nanoparticles), where the largest enhancement of the chemiluminescence response was observed on SIFs with high loading. These findings allowed us to demonstrate the use of SIFs (high loading) for the detection of a biologically relevant target protein (glial fibrillary acidic protein or GFAP), where the chemiluminescence response of the standard bioassay for GFAP was enhanced up to ~50% as compared to bioassay on glass slides. PMID:26582101

  2. Flow-through bioassay for measuring bioaccumulation of toxic substances from sediment

    USGS Publications Warehouse

    Mac, Michael J.; Edsall, Carol C.; Hesselberg, Robert J.; Sayers, Richard E., Jr.

    1984-01-01

    Over 10 million cubic meters of sediment are dredged annually from Great Lakes waterways. Because much of this material is taken from harbors, connecting channels, and other nearshore areas that often are contaminated with toxic substances, the sediments proposed for dredging need to be evaluated for the presence of bioavailable contaminants and the potential for toxicity to the biota. Sound decisions on the appropriate disposal of the dredged material can be made only after such an evaluation. Presently, no standardized procedure exists for evaluating dredged material in freshwater systems although current criteria for discharge of dredged material into marine water have been developed (USEPA/CE 1977). In the ocean discharge guideline, it is recommended that bioassays be conducted on liquid, solid, and suspended particulate phases of dredged material. because it appears that the solid phase has the greatest potential for environmental damage and because measurement of bioaccumulation must be made to evaluate sediments for disposal (USEPA/CE 1977, Seeyle and Mac 1983), we developed a bioassay for testing the solid phase of dredged material that measures the survival of organisms and, perhaps more important, the bioaccumulation of toxic substances by aquatic organisms from naturally contaminated sediments (Peddicord et al. 1980; Rubinstein et al. 1980, 1983; Seeyle st al. 1982), several have used testing methods that result in unacceptable mortality to control organisms (Bahnick et al. 1981, Prater et al. 1983). Our bioassay is intended to estimate the potential for bioaccumlation of contaminants from sediments that are not acutely toxic to test organisms, but are suspected of containing persistent contaminants. By using test organisms that are not highly susceptible to toxic compounds, the bioaccumulation test allows estimation of the potential food-chain accumulation of contaminants that may occur in local biota from surficial sediments. In practice

  3. A novel method for standardized application of fungal spore coatings for mosquito exposure bioassays

    PubMed Central

    2010-01-01

    Background Interest in the use of fungal entomopathogens against malaria vectors is growing. Fungal spores infect insects via the cuticle and can be applied directly on the insect to evaluate infectivity. For flying insects such as mosquitoes, however, application of fungal suspensions on resting surfaces is more realistic and representative of field settings. For this type of exposure, it is essential to apply specific amounts of fungal spores homogeneously over a surface for testing the effects of fungal dose and exposure time. Contemporary methods such as spraying or brushing spore suspensions onto substrates do not produce the uniformity and consistency that standardized laboratory assays require. Two novel fungus application methods using equipment developed in the paint industry are presented and compared. Methods Wired, stainless steel K-bars were tested and optimized for coating fungal spore suspensions onto paper substrates. Different solvents and substrates were evaluated. Two types of coating techniques were compared, i.e. manual and automated coating. A standardized bioassay set-up was designed for testing coated spores against malaria mosquitoes. Results K-bar coating provided consistent applications of spore layers onto paper substrates. Viscous Ondina oil formulations were not suitable and significantly reduced spore infectivity. Evaporative Shellsol T solvent dried quickly and resulted in high spore infectivity to mosquitoes. Smooth proofing papers were the most effective substrate and showed higher infectivity than cardboard substrates. Manually and mechanically applied spore coatings showed similar and reproducible effects on mosquito survival. The standardized mosquito exposure bioassay was effective and consistent in measuring effects of fungal dose and exposure time. Conclusions K-bar coating is a simple and consistent method for applying fungal spore suspensions onto paper substrates and can produce coating layers with accurate effective spore

  4. Development of marine sediment bioassays and toxicity tests for monitoring and regulation in Europe

    SciTech Connect

    Thain, J.; Matthiessen, P.

    1995-12-31

    There is a need in Europe and elsewhere for a broad suite of whole-sediment bioassays and toxicity tests which can be used for routine monitoring and assessment of the marine environment and for evaluating the toxic effects of chemicals which may find their way into sediments. Until recently, few European species had been incorporated into such tests but the availability of suitable methodologies is now increasing rapidly. Perhaps the most important recent activity in this area consisted of an international ring test of acute sediment toxicity test methods which was organized by the Oslo and Paris Commissions in 1993, using up to 4 offshore chemicals as test materials. It evaluated the performance of 4 acute (5--10 day) tests involving: the sea urchin Echinocardium cordatum, the bivalve mollusc Abra alba, the amphipod crustacean Corophium volutator, and the polychaete worm Arenicola marina. The ring test concluded that the C. volutator test was the most appropriate for evaluating offshore chemicals, but all these methods are now widely used in Europe, both as toxicity tests and as bioassays. For example, the A. marina procedure (which has both lethal and sublethal endpoints), in combination with the C. volutator method, is now routinely used in the UK for monitoring the toxicity of estuarine sediments. Further activities are in progress. Perhaps the most important is the development of chronic marine sediment tests and bioassays which can be used to assess the long-term effects of the many sedimentary contaminants which are able to persist in this type of habitat and possibly cause delayed effects on the growth and reproduction, etc. of benthic fauna.

  5. Application of a lux-based bioassay to assess soil toxicity

    SciTech Connect

    Paton, G.I. |; Campbell, C.D.; Rattray, E.A.S.; Glover, L.A.; Killham, K.

    1995-12-31

    The expression of prokaryotic bioluminescence is linked with cell metabolism and accordingly bioassays have been developed using naturally bioluminescent bacteria to assess ecotoxicity. Advances in biotechnology have allowed the isolation of the lux genes (responsible for bioluminescence) from marine organisms and their insertion into terrestrial bacteria. This has enabled the use of ecologically relevant bacteria to assess toxicity by measuring bioluminescence response in the presence of toxins. The lux genes were inserted into Pseudomonas fluorescens and Rhizobium leguminosarum biovar trifolii as a multi-copy plasmid and also integrated into the chromosome. It was found that in aqueous solutions the plasmid constructs were more sensitive than the chromosomal constructs to a range of toxins. The order of toxicity for Ps. fluorescens was Zn = Cu > Cd > Ni > Cr > DCP and for R. trifolii Zn > Cu > Cd > DCP > Cr. The lux based bioassays were more reproducible and sensitive than ATP and dehydrogenase assays and offered greater sensitivity than Photobacterium phosphoreum assays to assess toxicity of inorganic pollutants. Extracts from 4 soil types were spiked with a range of toxins and when EC{sub 50} values were determined it was shown that toxicity was related to soil characteristics. This enabled the assay to be used to assess the Lee Valley soil experiment which represents an important international study of the effect of the application of contaminated sewage to land. High metal application rates had been shown to have serious implications for soil ecology. Chemical analysis, carried out 26 years after sewage addition confirmed that soil extracts still had increased metal concentrations. The lux-based bioassays, which proved to be rapid, reproducible and sensitive confirmed that the metals were still biologically available and hence toxic.

  6. Fast and accurate semantic annotation of bioassays exploiting a hybrid of machine learning and user confirmation.

    PubMed

    Clark, Alex M; Bunin, Barry A; Litterman, Nadia K; Schürer, Stephan C; Visser, Ubbo

    2014-01-01

    Bioinformatics and computer aided drug design rely on the curation of a large number of protocols for biological assays that measure the ability of potential drugs to achieve a therapeutic effect. These assay protocols are generally published by scientists in the form of plain text, which needs to be more precisely annotated in order to be useful to software methods. We have developed a pragmatic approach to describing assays according to the semantic definitions of the BioAssay Ontology (BAO) project, using a hybrid of machine learning based on natural language processing, and a simplified user interface designed to help scientists curate their data with minimum effort. We have carried out this work based on the premise that pure machine learning is insufficiently accurate, and that expecting scientists to find the time to annotate their protocols manually is unrealistic. By combining these approaches, we have created an effective prototype for which annotation of bioassay text within the domain of the training set can be accomplished very quickly. Well-trained annotations require single-click user approval, while annotations from outside the training set domain can be identified using the search feature of a well-designed user interface, and subsequently used to improve the underlying models. By drastically reducing the time required for scientists to annotate their assays, we can realistically advocate for semantic annotation to become a standard part of the publication process. Once even a small proportion of the public body of bioassay data is marked up, bioinformatics researchers can begin to construct sophisticated and useful searching and analysis algorithms that will provide a diverse and powerful set of tools for drug discovery researchers. PMID:25165633

  7. Fast and accurate semantic annotation of bioassays exploiting a hybrid of machine learning and user confirmation

    PubMed Central

    Bunin, Barry A.; Litterman, Nadia K.; Schürer, Stephan C.; Visser, Ubbo

    2014-01-01

    Bioinformatics and computer aided drug design rely on the curation of a large number of protocols for biological assays that measure the ability of potential drugs to achieve a therapeutic effect. These assay protocols are generally published by scientists in the form of plain text, which needs to be more precisely annotated in order to be useful to software methods. We have developed a pragmatic approach to describing assays according to the semantic definitions of the BioAssay Ontology (BAO) project, using a hybrid of machine learning based on natural language processing, and a simplified user interface designed to help scientists curate their data with minimum effort. We have carried out this work based on the premise that pure machine learning is insufficiently accurate, and that expecting scientists to find the time to annotate their protocols manually is unrealistic. By combining these approaches, we have created an effective prototype for which annotation of bioassay text within the domain of the training set can be accomplished very quickly. Well-trained annotations require single-click user approval, while annotations from outside the training set domain can be identified using the search feature of a well-designed user interface, and subsequently used to improve the underlying models. By drastically reducing the time required for scientists to annotate their assays, we can realistically advocate for semantic annotation to become a standard part of the publication process. Once even a small proportion of the public body of bioassay data is marked up, bioinformatics researchers can begin to construct sophisticated and useful searching and analysis algorithms that will provide a diverse and powerful set of tools for drug discovery researchers. PMID:25165633

  8. Guidance document for prepermit bioassay testing of low-level radioactive waste

    SciTech Connect

    Anderson, S.L.; Harrison, F.L.

    1990-11-01

    In response to the mandate of Public Law 92-532, the Marine Protection, Research, and Sanctuaries Act (MPRSA) of 1972, as amended, the Environmental Protection Agency (EPA) has developed a program to promulgate regulations and criteria to control the ocean disposal of radioactive wastes. The EPA seeks to understand the mechanisms for biological response of marine organisms to the low levels of radioactivity that may arise from the release of these wastes as a result of ocean-disposal practices. Such information will play an important role in determining the adequacy of environmental assessments provided to the EPA in support of any disposal permit application. Although the EPA requires packaging of low-level radioactive waste to prevent release during radiodecay of the materials, some release of radioactive material into the deep-sea environment may occur when a package deteriorates. Therefore, methods for evaluating the impact on biota are being evaluated. Mortality and phenotypic responses are not anticipated at the expected low environmental levels that might occur if radioactive materials were released from the low-level waste packages. Therefore, traditional bioassay systems are unsuitable for assessing sublethal effects on biota in the marine environment. The EPA Office of Radiation Programs (ORP) has had an ongoing program to examine sublethal responses to radiation at the cellular level, using cytogenetic end points. This technical guidance report represents prepermit bioassay procedures that potentially may be applicable to the assessment of effects from a mixture of radionuclides that could be released from a point source at the ocean bottom. Methodologies along with rationale and a discussion of uncertainty are presented for the sediment benthic bioassay protocols identified in this report.

  9. Evaluation of the toxicity of marine sediments and dredge spoils with the MicrotoxR bioassay

    USGS Publications Warehouse

    Ankley, G.T.; Hoke, R.A.; Giesy, J.P.; Winger, P.V.

    1989-01-01

    The MicrotoxR bioassay was used to evaluate the toxicity of sediment and dredge spoil elutriates from several potentially-contaminated sites in Mobile and Pascagoula Bays. Elutriates were prepared using either local seawater or distilled deionized water (osmotically adjusted with NaCl prior to testing), and MicrotoxR assays were performed with the elutriates and three reference toxicants. There were marked differences in the toxicity of several elutriates and reference toxicants in the two different waters, with the seawater generally resulting in the same or lesser toxicity than the osmotically-adjusted distilled deionized water.

  10. BIOASSAY OF POLLUTED SEDIMENTS AND REDUCTION OF TOXICITY BY AEROBIC TREATMENT

    NASA Astrophysics Data System (ADS)

    Sumikura, Mitsuhiro; Kojima, Toshikazu; Okamura, Kazuo; Horiuchi, Sumio

    Aerobic treatment is being studied as an efficient in-situ remediation method for polluted sediments. This treatment method is able to decompose organic substances that are otherwise difficult to degrade. Changes in toxicity during such treatment is the subject of this study. Bioassay utilizing Daphnia magna was conducted for toxicity assessment of sediment. Laboratory treatment experiment was conducted, and changes in toxicity and dissolved ion concentrations were measured.Conclusions from this test are, as follows; (1) toxicity of chloride, ammonia, and sulfide was found to be masked by the coexisting materials in the sample matrix, and (2) changes of toxicity was dependent on the forms of sulfur and nitrogen species.

  11. Application of short-term bioassays to the assessment of engine exhaust emissions

    SciTech Connect

    Shore, P.R.; Tesh, J.M.; Bootman, J.

    1987-01-01

    Extracts of particulate emissions from light duty diesel, conventional gasoline and lean-burn gasoline engines have been analysed using a range of short-term bioassays. The intention of the analyses was to identify tests which could be routinely applied to exhaust assessment in order to study the effects of engine operating conditions and design on biological activity. In this respect the most promising assays were the Ames Salmonella typhimurium test for mutagenicity, the detection of chromosome damage in rat liver cells, and the assessment of growth and development defects in the Hydra Regeneration Assay.

  12. A Bayesian approach to the analysis of quantal bioassay studies using nonparametric mixture models.

    PubMed

    Fronczyk, Kassandra; Kottas, Athanasios

    2014-03-01

    We develop a Bayesian nonparametric mixture modeling framework for quantal bioassay settings. The approach is built upon modeling dose-dependent response distributions. We adopt a structured nonparametric prior mixture model, which induces a monotonicity restriction for the dose-response curve. Particular emphasis is placed on the key risk assessment goal of calibration for the dose level that corresponds to a specified response. The proposed methodology yields flexible inference for the dose-response relationship as well as for other inferential objectives, as illustrated with two data sets from the literature. PMID:24354490

  13. Promising Aedes aegypti Repellent Chemotypes Identified through Integrated QSAR, Virtual Screening, Synthesis, and Bioassay

    PubMed Central

    Oliferenko, Polina V.; Oliferenko, Alexander A.; Poda, Gennadiy I.; Osolodkin, Dmitry I.; Pillai, Girinath G.; Bernier, Ulrich R.; Tsikolia, Maia; Agramonte, Natasha M.; Clark, Gary G.; Linthicum, Kenneth J.; Katritzky, Alan R.

    2013-01-01

    Molecular field topology analysis, scaffold hopping, and molecular docking were used as complementary computational tools for the design of repellents for Aedes aegypti, the insect vector for yellow fever, chikungunya, and dengue fever. A large number of analogues were evaluated by virtual screening with Glide molecular docking software. This produced several dozen hits that were either synthesized or procured from commercial sources. Analysis of these compounds by a repellent bioassay resulted in a few highly active chemicals (in terms of minimum effective dosage) as viable candidates for further hit-to-lead and lead optimization effort. PMID:24039693

  14. Discovery of active components in herbs using chromatographic separation coupled with online bioassay.

    PubMed

    De-Qiang, Li; Zhao, Jing; Wu, Dong; Shao-Ping, Li

    2016-05-15

    Discovery of bioactive compounds from complex mixtures is a challenge. In past decades, several strategies were developed and implemented for rapid and effective screening and characterization of bioactive components in complex matrices. This review mainly focused on the online strategies, which integrated the separation science, mass spectrometry, and bioactivity screening in a single platform, allowing simultaneous screening and characterization of active compounds from complex matrices, especially from the herbs. The online screening methodologies, including pre-column affinity-based screening and post-column bioassay, were discussed and their applied examples were also presented to illustrate the strengths and limitations of these approaches. PMID:26896311

  15. Principles for the selection of doses in chronic rodent bioassays. ILSI Risk Science Working Group on Dose Selection.

    PubMed Central

    Foran, J A

    1997-01-01

    Dose selection in chronic rodent bioassays has been one of the most debated issues in risk assessment. The Committee on Risk Assessment Methods of the National Research Council attempted, but failed, in 1993 to reach consensus on how to select doses for chronic rodent bioassays. However, a more recent effort conducted by the ILSI Risk Science Institute has resulted in a consensus set of principles for dose selection, including selection of the highest dose for chronic rodent bioassays. The principles encourage a move away from sole reliance on a maximum tolerated dose (MTD), as it has been traditionally defined (primarily by body weight and histopathology), and toward the use of sound scientific and toxicologic principles for the selection of all doses in the chronic bioassay. Specifically, the principles recommend that dose selection for chronic studies must be based on sound toxicologic principles; dose selection should consider human exposure; dose selection should be based on a variety of endpoints and effects derived from prechronic studies; and dose selection should consider physicochemical and other factors. Implementation of the principles internationally will have two important benefits; improvement in the quality and consistency of the rodent bioassay and international harmonization of dose selection procedures. PMID:9074868

  16. Estimating Prion Adsorption Capacity of Soil by BioAssay of Subtracted Infectivity from Complex Solutions (BASICS)

    PubMed Central

    Wyckoff, A. Christy; Lockwood, Krista L.; Meyerett-Reid, Crystal; Michel, Brady A.; Bender, Heather; VerCauteren, Kurt C.; Zabel, Mark D.

    2013-01-01

    Prions, the infectious agent of scrapie, chronic wasting disease and other transmissible spongiform encephalopathies, are misfolded proteins that are highly stable and resistant to degradation. Prions are known to associate with clay and other soil components, enhancing their persistence and surprisingly, transmissibility. Currently, few detection and quantification methods exist for prions in soil, hindering an understanding of prion persistence and infectivity in the environment. Variability in apparent infectious titers of prions when bound to soil has complicated attempts to quantify the binding capacity of soil for prion infectivity. Here, we quantify the prion adsorption capacity of whole, sandy loam soil (SLS) typically found in CWD endemic areas in Colorado; and purified montmorillonite clay (Mte), previously shown to bind prions, by BioAssay of Subtracted Infectivity in Complex Solutions (BASICS). We incubated prion positive 10% brain homogenate from terminally sick mice infected with the Rocky Mountain Lab strain of mouse-adapted prions (RML) with 10% SLS or Mte. After 24 hours samples were centrifuged five minutes at 200×g and soil-free supernatant was intracerebrally inoculated into prion susceptible indicator mice. We used the number of days post inoculation to clinical disease to calculate the infectious titer remaining in the supernatant, which we subtracted from the starting titer to determine the infectious prion binding capacity of SLS and Mte. BASICS indicated SLS bound and removed ≥ 95% of infectivity. Mte bound and removed lethal doses (99.98%) of prions from inocula, effectively preventing disease in the mice. Our data reveal significant prion-binding capacity of soil and the utility of BASICS to estimate prion loads and investigate persistence and decomposition in the environment. Additionally, since Mte successfully rescued the mice from prion disease, Mte might be used for remediation and decontamination protocols. PMID:23484043

  17. Using marine bioassays to classify the toxicity of Dutch harbor sediments.

    PubMed

    Stronkhorst, Joost; Schipper, Cor; Brils, Jos; Dubbeldam, Marco; Postma, Jaap; van de Hoeven, Nelly

    2003-07-01

    A procedure was developed to assess contaminated marine sediments from Dutch harbors for possible adverse biological effects using three laboratory bioassays: A 10-d survival test with the amphipod Corophium volutator, a 14-d survival test with the heart urchin Echinocardium cordatum (adults), and the bioluminescence inhibition test with the bacterium Vibrio fischeri (Microtox solid phase test LSP]). Microtox results were mathematically corrected for the modifying influence of fine sediment particles. After a validation procedure on test performance and modifying factors, respectively, 81%, 99%, and 90% of the amphipod, heart urchin, and Microtox results were approved. Lower and upper threshold limits for biological effects were set at respectively 24 and 30% mortality for C. volutator, 27 and 35% mortality for E. cordatum, and 24 and 48 toxic units for the Microtox SP based on significant differences with control sediment and the performance of reference sediments. The bioassays clearly distinguished harbor sediments that give rise to acute effects and those that do not. Threshold limits for the amphipods, heart urchins, and bacteria were exceeded in, respectively, 9 to 17%, 33 to 40%, and 23 to 50% of the sediment samples. Highest effects were observed in sediments from the northerly harbors; there was significantly less response in sediments from the Delta Region and the port of Rotterdam (The Netherlands). The procedure outlined in this paper can be used for routine screening of contaminated dredged material that is proposed for open water disposal. PMID:12836979

  18. Evaluation of bioassays to monitor surface microlayer toxicity in tropical marine waters.

    PubMed

    Rumbold, D G; Snedaker, S C

    1997-02-01

    Bioassays were developed, using embryos of: coral,Montastraea faveolata; graysby, Epinephelus cruentatus;grouper, Epinephelus adscensionis x gruttatus (hybrid); queenconch, Strombus gigas; rock-boring urchin, Echinodermatalucunter; spotted seatrout, Cynoscion nebulosus; variegatedurchin, Lytechinus variegatus; winged pearl oyster, Pteriacolymbus; and yellowtail snapper, Ocyurus chrysurus. Relativesensitivities and precison of various species-endpoint combinations wereevaluated using three reference toxicants: copper, sodium dodecyl sulfate,and Dibrom(R). The 24-h P. colymbus embryo test had the best overallsensitivity and exhibited a high degree of precision. However, oyster embryoswere difficult to obtain and did not aggregate at the air-water interface.Therefore, the P. colymbus embryo test was deemed unsuitable for useas a bioassay for monitoring sea-surface microlayer (SSML) toxicity. Testsbased on normal development of L. variegatus to the early pluteus 3stage and percent normal-live C. nebulosus larvae at 48 h wererelatively sensitive and exhibited good replicability and repeatability. TheL. variegatus urchin embryo test was also found to be highlyreproducible. The results of this comparative study indicated that L.variegatus and C. nebulosus were suitable surrogates forcoral-reef species in toxicity assessments of the SSML. PMID:9069187

  19. Environmental effects of dredging: Upland animal bioassays of dredged material. Technical notes

    SciTech Connect

    Stafford, E.A.; Edwards, C.A.

    1988-02-01

    Earthworms have great potential for use as bioassay/biomonitor organisms in studies of contaminant uptake and possess many characteristics that make them ideally suited for this purpose (Ma 1982). Studies have demonstrated that native species of earthworms, collected at contaminated sites, can be used to indicate biologically available levels of these contaminants (Helmke et al. 1979, Ireland 1983, Pietz et al. 1984). However, it is the species Eisenia foetida (which does not naturally colonize these sites) which has been recommended for use in the laboratory for the ecotoxicological testing of agricultural and industrial chemicals (European Economic Community (EEC) 1984), proposed as a bioassay species for assessing contaminant availability in waste materials, and used to determine the bioavallability of contaminants in dredged material (Marquenie and Simmers 1984). Correlations between total and OTPA-extractable metal concentrations in contaminated substrates and the concentrations in the tissues of earthworms exposed to these substrates over a 28-day period may be used to establish their potential as biomonitor organisms.

  20. CALUX bioassay: a cost-effective rapid screening technique for screening dioxins like compounds.

    PubMed

    Sakthivel, Selvaraj; Balasubramanian, Prithiviraj; Nakamura, Masafumi; Ko, Shunkei; Chakraborty, Paromita

    2016-03-01

    Xenobiotic detection systems-chemically activated luciferase expression (XDS-CALUX) bioassay in determining the toxic equivalency (TEQ) of PCDD/Fs from contaminated sites reported in several papers has been discussed in this study. CALUX bioassay method has been validated by an effective combined column clean-up system followed by addition of samples to monolayer cell cultures of H1L6.1c3 cell line in 96 well plates. Cultures are then examined under microscope after 24 h incubation followed by rinsing with 75 μL phosphate buffer saline and 30 μL of cell culture lysis. The response is observed in the luminometer and expressed in relative light unit (RLUs). CALUX-TEQ is estimated from a TCDD standard curve for unknown samples. Quality control in CALUX is done by selecting the range of CALUX values falling in the center of the linear standard curve. For developing nations CALUX biossay can be used as a cost effective and rapid screening technique for screening xenobiotic compounds from the hotspots like open solid waste burning sites, informal e-waste recycling workshops and industrial zones where constant monitoring for such compounds is required. PMID:26943601

  1. Potential risk of biochar-amended soil to aquatic systems: an evaluation based on aquatic bioassays.

    PubMed

    Bastos, A C; Prodana, M; Abrantes, N; Keizer, J J; Soares, A M V M; Loureiro, S

    2014-11-01

    It is vital to address potential risks to aquatic ecosystems exposed to runoff and leachates from biochar-amended soils, before large scale applications can be considered. So far, there are no established approaches for such an assessment. This study used a battery of bioassays and representative aquatic organisms for assessing the acute toxicity of water-extractable fractions of biochar-amended soil, at reported application rates (80 t ha(-1)). Biochar-amended aqueous soil extracts contained cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), manganese (Mn), zinc (Zn), nickel (Ni), lead (Pb), arsenic (As) and mercury (Hg) (Σmetals 96.3 µg l(-1)) as well as the 16 priority PAHs defined by the U.S. Environmental Protection Agency (Σ16PAHs 106 ng l(-1)) at contents in the range of current EU regulations for surface waters. Nevertheless, acute exposure to soil-biochar (SB) extracts resulted in species-specific effects and dose-response patterns. While the bioluminescent marine bacterium Vibrio fischeri was the most sensitive organism to aqueous SB extracts, there were no effects on the growth of the microalgae Pseudokirchneriella subcapitata. In contrast, up to 20 and 25% mobility impairment was obtained for the invertebrate Daphnia magna upon exposure to 50 and 100% SB extract concentrations (respectively). Results suggest that a battery of rapid and cost-effective aquatic bioassays that account for ecological representation can complement analytical characterization of biochar-amended soils and risk assessment approaches for surface and groundwater protection. PMID:25213286

  2. Bioassays of Bacillus sphaericus (strain 1593) against mosquitoes of public health importance in Malaysia.

    PubMed

    Cheong, W C; Yap, H H

    1985-03-01

    The pathogenicity of Bacillus sphaericus strain 1593 was tested against laboratory-reared larvae of four local species of mosquitoes of public health importance in Malaysia; Aedes aegypti, Anopheles balabacensis, Mansonia uniformis and Culex quinquefasciatus. The bacteria was shake-cultured at 28 +/- 1 degrees C for three days, using Glucose-Yeast Extract Salts medium. After which, the spores and vegetative cells were harvested and stored at 4 degrees C before use. Conditions for bioassays were mean temperature of 25 +/- 1 degrees C and relative humidity 65 +/- 5.0. Twenty third-instar larvae of each species were assayed in 90 ml of diluted spore solution. Each concentration and a control were replicated three times for each bioassay. Larval mortalities at 24 hours and 48 hours were taken and analyzed through Probit Analysis using a computer (IBM 370). LC50 values after 48 hours of exposure showed an increasing order of larval susceptibility as follows: Ae. aegypti (417.70 x 10(4)), An. balabacensis (45.84 x 10(4)), Ma. uniformis (18.23 x 10(4)) and Cx. quinquefasciatus (4.14 x 10(4) spores/ml). With the ability to kill 90% of the Cx. quinquefasciatus larvae tested with just a concentration of 10(5) spores/ml, B. sphaericus (strain 1593) has shown good potential as a biocontrol agent for this species of mosquito. PMID:4023816

  3. Botulinum Neurotoxin Serotype a Specific Cell-Based Potency Assay to Replace the Mouse Bioassay

    PubMed Central

    Fernández-Salas, Ester; Wang, Joanne; Molina, Yanira; Nelson, Jeremy B.; Jacky, Birgitte P. S.; Aoki, K. Roger

    2012-01-01

    Botulinum neurotoxin serotype A (BoNT/A), a potent therapeutic used to treat various disorders, inhibits vesicular neurotransmitter exocytosis by cleaving SNAP25. Development of cell-based potency assays (CBPAs) to assess the biological function of BoNT/A have been challenging because of its potency. CBPAs can evaluate the key steps of BoNT action: receptor binding, internalization-translocation, and catalytic activity; and therefore could replace the current mouse bioassay. Primary neurons possess appropriate sensitivity to develop potential replacement assays but those potency assays are difficult to perform and validate. This report describes a CBPA utilizing differentiated human neuroblastoma SiMa cells and a sandwich ELISA that measures BoNT/A-dependent intracellular increase of cleaved SNAP25. Assay sensitivity is similar to the mouse bioassay and measures neurotoxin biological activity in bulk drug substance and BOTOX® product (onabotulinumtoxinA). Validation of a version of this CBPA in a Quality Control laboratory has led to FDA, Health Canada, and European Union approval for potency testing of BOTOX®, BOTOX® Cosmetic, and Vistabel®. Moreover, we also developed and optimized a BoNT/A CBPA screening assay that can be used for the discovery of novel BoNT/A inhibitors to treat human disease. PMID:23185348

  4. Assessment of the environmental quality of coastal sediments by using a combination of in vitro bioassays.

    PubMed

    Pérez-Albaladejo, Elisabet; Rizzi, Juliane; Fernandes, Denise; Lille-Langøy, Roger; Karlsen, Odd André; Goksøyr, Anders; Oros, Andra; Spagnoli, Federico; Porte, Cinta

    2016-07-15

    The environmental quality of marine sediments collected in the area of influence of the Po and Danube Rivers was assessed by using a battery of bioassays based on the use of PLHC-1 cells, zebrafish-Pxr-transfected COS-7 cells, and sea bass ovarian subcellular fractions. This allowed the determination of multiple endpoints, namely, cytotoxicity, oxidative stress, induction of CYP1A, activation of zebrafish Pxr and inhibition of ovarian aromatase. Organic extracts of sediments influenced by the Danube River and collected near harbors and urban discharges showed significant cytotoxicity, CYP1A induction and inhibition of aromatase activity. An analogous response of CYP1A induction and zfPxr activation was observed, which suggests the existence of common ligands of AhR and PXR in the sediment extracts. The study highlights the usefulness of the selected bioassays to identify those sediments that could pose a risk to aquatic organisms and that require further action in order to improve their environmental quality. PMID:27207027

  5. Automated cytochrome c oxidase bioassay developed for ionic liquids' toxicity assessment.

    PubMed

    Costa, Susana P F; Martins, Bárbara S F; Pinto, Paula C A G; Saraiva, M Lúcia M F S

    2016-05-15

    A fully automated cytochrome c oxidase assay resorting to sequential injection analysis (SIA) was developed for the first time and implemented to evaluate potential toxic compounds. The bioassay was validated by evaluation of 15 ionic liquids (ILs) with distinct cationic head groups, alkyl side chains and anions. The assay was based on cytochrome c oxidase activity reduction in presence of tested compounds and quantification of inhibitor concentration required to cause 50% of enzyme activity inhibition (EC50). The obtained results demonstrated that enzyme activity was considerably inhibited by BF4 anion and ILs incorporating non-aromatic pyrrolidinium and tetrabutylphosphonium cation cores. Emim [Ac] and chol [Ac], on contrary, presented the higher EC50 values among the ILs tested. The developed automated SIA methodology is a simple and robust high-throughput screening bioassay and exhibited good repeatability in all the tested conditions (rsd<3.7%, n=10). Therefore, it is expected that due to its simplicity and low cost, the developed approach can be used as alternative to traditional screening assays for evaluation of ILs toxicity and identification of possible toxicophore structures. Additionally, the results presented in this study provide further information about ILs toxicity. PMID:26894289

  6. Comparative study of three oligochaete species as indicators of metals in a sediment toxicity bioassay

    SciTech Connect

    Chapman, K.; Scheuerman, P.; Lanza, G.; Nelson, D.; Brinkhurst, R.

    1995-12-31

    Three oligochaete species, Tubifex tubifex, Branchiura sowerbyi and Lumbriculus variegatus, were analyzed for bioaccumulation and reproductive effects from reference sediment spiked with Cd or Cu. Sediment was spiked using the Sediment Suspension method to achieve concentrations of 4.0, 8.0 and 16.0 mg Cd/kg sediment (dry weight) and 25.0, 36.0, 50.0, 100.0 mg Cu/kg sediment (dry weight) . The bioassay was conducted under aerated, static conditions for 28 d at 22.5 C. Reproductive effects consisting of number of cocoons and eggs produced a negative linear regression with increasing Cd concentration. Cocoon volume remained consistent. Cu was more toxic to T. tubifex in this bioassay than results reported by the USEPA using similar concentrations. Lower concentrations of Cu also showed a negative linear regression with reproductive effects showing that oligochaetes could be a feasible indicator organism for sediment toxicity in a standardized ecological impact assay using reproduction as an endpoint.

  7. A review of whole animal bioassays of the carcinogenic potential of naphthalene.

    PubMed

    North, D Warner; Abdo, Kamal M; Benson, Janet M; Dahl, Alan R; Morris, John B; Renne, Roger; Witschi, Hanspeter

    2008-07-01

    This report provides a summary of deliberations conducted under the charge for members of Module A participating in the Naphthalene State-of-the-Science Symposium (NS3), Monterey, CA, October 9-12, 2006. Whole animal bioassays have been performed by the National Toxicology Program in mice and rats to ascertain the carcinogenic potential of naphthalene by inhalation exposure. A statistically significant increased incidence of pulmonary alveolar/bronchiolar adenoma (a benign lesion), was observed among female mice; an observed increase among the males did not reach statistical significance. No nasal tumors were observed in either sex. A tumorigenic response was observed in both sexes of rats, in males an increased incidence of nasal respiratory epithelium adenoma (a benign rather than malignant lesion) and in females, olfactory epithelial neuroblastoma. Interpretations of these studies vary. On the one hand, evidence of extensive non-neoplastic response in both sexes of both species indicates cytotoxicity occurred at all doses, and strongly suggests that cytotoxicity played a significant role in the tumor responses observed in the target tissues. On the other hand, olfactory epithelial neuroblastoma has rarely been observed in NTP bioassays. This review seeks to develop a consensus understanding of the scientific evidence provided by these studies, taking into account that they have been used as the basis for quantitative human cancer risk assessment, and suggests scientific studies that, if performed, could resolve scientific uncertainties. PMID:18364246

  8. Antimicrobial Activity and Brine Shrimp Lethality Bioassay of the Leaves Extract of Dillenia indica Linn

    PubMed Central

    Apu, AS; Muhit, MA; Tareq, SM; Pathan, AH; Jamaluddin, ATM; Ahmed, M

    2010-01-01

    The crude methanolic extract of Dillenia indica Linn. (Dilleniaceae) leaves has been investigated for the evaluation of antimicrobial and cytotoxic activities. Organic solvent (n-hexane, carbon tetrachloride and chloroform) fractions of methanolic extract and methanolic fraction (aqueous) were screened for their antimicrobial activity by disc diffusion method. Besides, the fractions were screened for cytotoxic activity using brine shrimp (Artemia salina) lethality bioassay. Among the four fractions tested, n-hexane, carbon tetrachloride, and chloroform fractions showed moderate antibacterial and antifungal activity compared to standard antibiotic, kanamycin. The average zone of inhibition was ranged from 6 to 8 mm at a concentration of 400 µg/disc. But the aqueous fraction was found to be insensitive to microbial growth. Compared to vincristine sulfate (with LC50 of 0.52 µg/ ml), n-hexane and chloroform fractions demonstrated a significant cytotoxic activity (having LC50 of 1.94 µg/ml and 2.13 µg/ml, respectively). The LC50 values of the carbon tetrachloride and aqueous fraction were 4.46 µg/ml and 5.13 µg/ ml, respectively. The study confirms the moderate antimicrobial and potent cytotoxic activities of Dillenia indica leaves extract and therefore demands the isolation of active principles and thorough bioassay. PMID:21331191

  9. DOSEXPRT: A bioassay dosimetry code for Martin Marietta Energy Systems, Inc.

    SciTech Connect

    Ward, R.C.; Eckerman, K.F.

    1992-04-01

    The bioassay code DOSEXPRT was developed for Martin Marietta Energy Systems, Inc., to provide compliance with Department of Energy (DOE) Order 5480, Chapter 11. DOSEXPRT computes the intake of a radionuclide in any year (considering both acute and chronic intakes) from in vivo measurements of the retained activity and/or measurements of the activity in excreta. The committed effective and organ doses for the intake are computed as well as the effective and organ doses expected to be received in each calendar year out to 50 years beyond the year of intake. The bioassay records used as input for DOSEXPRT are extracted from the Martin Marietta Energy Systems Occupational Health Information System (OHIS). DOSEXPRT implements a set of algorithms with parameters governing the translocation, retention, and excretion of the nuclide contained in data files specific to the nuclide. These files also contain dose-per-unit-intake coefficients used to compute the committed dose equivalent for the intakes in the year. Annual organ and effective doses are computed using additional dose-rate files that contain data on the dose rate at various times following a unit intake. If measurements are presented for more than one assay for a given nuclide, DOSEXPRT estimates the intake by applying weights assigned in the nuclide file for each assay. DOSEXPRT is accessed off the OHIS MENU No. 4 and designed to be run as a batch processor, but can also be run interactively for testing purposes.

  10. Bioassay and characterization of soil microorganisms involved in the biodegradation of the fungicide, metalaxyl

    SciTech Connect

    Bailey, A.M.

    1985-01-01

    A sensitive bioassay was developed to detect low concentrations of metalaxyl in soils. The quantitative estimation of metalaxyl in soils was based on a significant positive relationship between the radial growth of Phytophthora boehmeriae and the log concentration of the fungicide in the agar. The isolate of P. boehmeriae was chosen for its sensitivity to metalaxyl as manifested in a linear growth response on cornmeal agar over a range of 2 to 30 ng/ml. The sensitivity and quantitative nature of the bioassay was confirmed by comparison with data obtained by using /sup 14/C-metalaxyl. Metabolism of metalaxyl was detected in three of five avocado soils that had repeated applications of the fungicide over 2-5 yr. The average disappearance of metalaxyl was 28 days, and in the most active soils was 14 days. The composition and level of the microbial populations of soils, either active or inactive in the breakdown of metalaxyl, did not differ. Fungal and bacterial microflora recovered from these two soils by use of either selective media or filtration techniques were capable of metabolizing metalaxyl over a 45-day period.

  11. Antiplasmodial Properties and Bioassay-Guided Fractionation of Ethyl Acetate Extracts from Carica papaya Leaves

    PubMed Central

    Melariri, Paula; Campbell, William; Etusim, Paschal; Smith, Peter

    2011-01-01

    We investigated the antiplasmodial properties of crude extracts from Carica papaya leaves to trace the activity through bioassay-guided fractionation. The greatest antiplasmodial activity was observed in the ethyl acetate crude extract. C. papaya showed a high selectivity for P. falciparum against CHO cells with a selectivity index of 249.25 and 185.37 in the chloroquine-sensitive D10 and chloroquine-resistant DD2 strains, respectively. Carica papaya ethyl acetate extract was subjected to bioassay-guided fractionation to ascertain the most active fraction, which was purified and identified using high-pressure liquid chromatography (HPLC) and GC-MS (Gas chromatography-Mass spectrometry) methods. Linoleic and linolenic acids identified from the ethyl acetate fraction showed IC50 of 6.88 μg/ml and 3.58 μg/ml, respectively. The study demonstrated greater antiplasmodial activity of the crude ethyl acetate extract of Carica papaya leaves with an IC50 of 2.96 ± 0.14 μg/ml when compared to the activity of the fractions and isolated compounds. PMID:22174990

  12. Bioassay-Guided Isolation of Two Flavonoids from Derris scandens with Topoisomerase II Poison Activity.

    PubMed

    Sangmalee, Suphattra; Laorpaksa, Areerat; Sritularak, Boonchoo; Sukrong, Suchada

    2016-04-01

    Derris scandens (ROXB.) BENTH. (Fabaceae) is used as an alternative treatment for cancer in Thai traditional medicine. Investigation of the topoisomerase II (Top2) poison of compounds isolated from this plant may reveal new drug leads for the treatment of cancer. Bioassay-guided isolation was performed on an extract of D. scandens stems using a yeast cell-based assay. A yeast strain expressing the top2-1 temperature-sensitive mutant was used to assay Top2 activity. At the permissive temperature of 25°C, yeast cells were highly sensitive to Top2 poison agents. At the semi-permissive temperature of 30°C, where enzyme activity was present but greatly diminished, cells displayed only marginal sensitivity. The bioassay-guided fractionation of the extract led to the isolation of two known isoflavones: 5,7,4'-trihydroxy-6,8-diprenylisoflavone (1) and lupalbigenin (2). These two compounds also displayed cytotoxicity against three different cancer cell lines, KB, MCF-7 and NCI-H187. In conclusion, Top2 poison agents from D. scandens are reported for the first time, substantiating the use of D. scandens in Thai traditional medicine for cancer treatment. PMID:26754253

  13. Metal toxicity and biodiversity in serpentine soils: application of bioassay tests and microarthropod index.

    PubMed

    Visioli, Giovanna; Menta, Cristina; Gardi, Ciro; Conti, Federica Delia

    2013-01-01

    Eco-toxicological or bioassay tests have been intensively discussed as tools for the evaluation of soil quality. Tests using soil organisms, including microarthropods and plants, allow direct estimates to be made of important soil characteristics and functions. In this study we compared the results obtained by two in vitro standard bioassays following ISO or OECD guidelines: (i) the short term-chronic phytotoxicity germination and root elongation test using three different plant species Cucumis sativus L. (Cucurbitaceae), Lepidium sativum L. (Brassicaceae), and Medicago sativa L. (Fabaceae) and (ii) the inhibition of reproduction of Folsomia candida (Collembola) by soil pollutants to investigate the toxicity of a serpentine soil present in the Italian Apennines, rich in heavy metals such as Ni, Cr, and Co. In addition, microarthropod communities were characterised to evaluate the effects of metal contents on the soil fauna in natural conditions. Abundances, Acari/Collembola ratio, biodiversity indices and the QBS-ar index were calculated. Our results demonstrate that the two in vitro tests distinguish differences correlated with metal and organic matter contents in four sub-sites within the serpentinite. Soil fauna characterisation, not previously performed on serpentine soils, revealed differences in the most vulnerable and adapted groups of microarthropods to soil among the four sub-sites: the microarthropod community was found to be rich in term of biodiversity in the sub-site characterised by a lower metal content and a higher organic matter content and vegetation. PMID:23107056

  14. Studies on the use of cultured cells in a bioassay for parathyroid hormone.

    PubMed

    Armston, A E; Wood, P J

    1994-11-01

    Measurement of parathyroid hormone (PTH) is important for diagnosing hyper- and hypoparathyroidism. The move to two-site immunometric assays that detect the whole molecule has improved the discrimination of these conditions but these assays may be too restrictive because some PTH fragments that are biologically active may not be detected. In addition, PTH-like peptide of malignancy, an important cause of malignancy-associated hypercalcaemia, is not detected by the two-site assays. Experiments were performed to set up a simple, robust and inexpensive bioassay for PTH, exploiting a kidney cell line and using cyclic AMP or an eluted stain assay as the end point. Of the 12 cell lines tested, an opossum kidney (WOK) cell line showed the most promise. Despite optimization of the procedure to include pre-treatment with dexamethasone, insulin and PTH, followed by incubation in the presence of 5'-guanylimidodiphosphate, isobutyl-1-methylxanthine and forskolin, the WOK cells showed insufficient sensitivity for use in a cultured cell bioassay for PTH in human serum. In addition, the cells were less sensitive to PTH-like peptide precluding their use for an assay for this molecule. PMID:7829991

  15. Bioavailability of iron from spinach using an in vitro/human Caco-2 cell bioassay model

    NASA Technical Reports Server (NTRS)

    Rutzke, Corinne J.; Glahn, Raymond P.; Rutzke, Michael A.; Welch, Ross M.; Langhans, Robert W.; Albright, Louis D.; Combs, Gerald F Jr; Wheeler, Raymond M.

    2004-01-01

    Spinach (Spinacia oleracea) cv Whitney was tested for iron bioavailabilty using an in vitro human intestinal cell culture ferritin bioassay technique previously developed. Spinach was cultured in a growth chamber for 33 days, harvested, and freeze-dried. Total iron in the samples was an average of 71 micrograms/g dry weight. Spinach was digested in vitro (pepsin and 0.1 M HCl followed by pancreatin and 0.1 M NaHCO3) with and without the addition of supplemental ascorbic acid. Caco-2 cell cultures were used to determine iron bioavailability from the spinach mixtures. Production of the iron-binding protein ferritin in the Caco-2 cells showed the supplemental ascorbic acid doubled bioavailability of iron from spinach. The data show fresh spinach is a poor source of iron, and emphasize the importance of evaluation of whole meals rather than single food items. The data support the usefulness of the in vitro/Caco-2 cell ferritin bioassay model for prescreening of space flight diets for bioavailable iron.

  16. DOSEXPRT: A bioassay dosimetry code for Martin Marietta Energy Systems, Inc

    SciTech Connect

    Ward, R.C.; Eckerman, K.F.

    1992-04-01

    The bioassay code DOSEXPRT was developed for Martin Marietta Energy Systems, Inc., to provide compliance with Department of Energy (DOE) Order 5480, Chapter 11. DOSEXPRT computes the intake of a radionuclide in any year (considering both acute and chronic intakes) from in vivo measurements of the retained activity and/or measurements of the activity in excreta. The committed effective and organ doses for the intake are computed as well as the effective and organ doses expected to be received in each calendar year out to 50 years beyond the year of intake. The bioassay records used as input for DOSEXPRT are extracted from the Martin Marietta Energy Systems Occupational Health Information System (OHIS). DOSEXPRT implements a set of algorithms with parameters governing the translocation, retention, and excretion of the nuclide contained in data files specific to the nuclide. These files also contain dose-per-unit-intake coefficients used to compute the committed dose equivalent for the intakes in the year. Annual organ and effective doses are computed using additional dose-rate files that contain data on the dose rate at various times following a unit intake. If measurements are presented for more than one assay for a given nuclide, DOSEXPRT estimates the intake by applying weights assigned in the nuclide file for each assay. DOSEXPRT is accessed off the OHIS MENU No. 4 and designed to be run as a batch processor, but can also be run interactively for testing purposes.

  17. Tradescantia-micronucleus (Trad-MCN) bioassay on clastogenicity of wastewater and in situ monitoring.

    PubMed

    Ruiz, E F; Rabago, V M; Lecona, S U; Perez, A B; Ma, T H

    1992-11-01

    The Tradescantia-micronucleus (Trad-MCN) bioassay was used to determine the clastogenicity of wastewater samples collected from the Arena canal which contains effluent from the industrial district Benito Juarez of the city of Queretaro, Mexico. Fifteen wastewater samples which were collected, in most cases, at bi-weekly intervals beginning in September 1986 through February 1988, after a 3-fold dilution were used to treat Tradescantia plant cuttings. The clastogenicity expressed in terms of micronucleus frequencies of treated groups (30 h of treatment without recovery time) was significantly (0.01) higher than that of the tapwater control groups. The Trad-MCN bioassay was also used for in situ monitoring of air pollutants for the clastogenicity at 3 sites near the industrial and residential areas (Flores Magon, Conalep and Bellas Artes) of the city of Queretaro. Fourteen monitoring trips were made to each of the 3 sites at monthly intervals beginning in May 1988 through June 1990. Seasonal variation of micronucleus frequencies was exhibited with the peak clastogenicities shown in May and June 1988, June 1989 and April 1990 at the three sites. Micronucleus frequencies of all the exposed groups at the Conalep site, a predominantly industrial area, were markedly higher than that of the laboratory control groups throughout the 2-year period. PMID:1383722

  18. Dose selection as it pertains to testing a prodrug in carcinogenicity bioassays.

    PubMed

    Szczech, G M; Tucker, W E

    1983-01-01

    Toxicologists need more information than is usually available in the early stages of development of a drug in order to choose proper dose levels for testing in the bioassays. The approach most likely to result in successful bioassays involves an early multidisciplinary effort in which there is pharmacokinetic characterization of the test material in both rats and mice. Preliminary 3 month studies are desirable. Periodic sampling of plasma is essential to detect possible non-linear kinetics (as in the example we report herein) reflected as accumulation of the test material or metabolites. This is true regardless of the test substance. However, if one tests prodrugs it may be particularly helpful to know if chemical or enzymatic conversion of the prodrug is linear and if there is reversion to prodrug or other abberant metabolism. Failure to rule out these possibilities could result in subsequent clinically irrelevant organ damage or could compromise longevity or the interpretation of results in lifetime studies. Pharmacokinetic considerations are as valid as the more traditional biologic or morphologic end points used to estimate maximum tolerated or no-effect dose levels. PMID:6681397

  19. A battery of bioassays for the evaluation of phenanthrene biotoxicity in soil.

    PubMed

    Khan, Muhammad Imran; Cheema, Sardar Alam; Tang, Xianjin; Hashmi, Muhammad Zaffar; Shen, Chaofeng; Park, Joonhong; Chen, Yingxu

    2013-07-01

    A battery of bioassays was used to assess the ecotoxicological risk of soil spiked with a range of phenanthrene levels (0.95, 6.29, 38.5, 58.7, 122, and 303 μg g(-1) dry soil) and aged for 69 days. Multiple species (viz. Brassica rapa, Eisenia feotida, Vibrio fischeri), representing different trophic levels, were used as bioindicator organisms. Among acute toxicity assays tested, the V. fischeri luminescence inhibition assay was the most sensitive indicator of phenanthrene biotoxicity. More than 15 % light inhibition was found at the lowest phenanthrene level (0.95 μg g(-1)). Furthermore, comet assay using E. fetida was applied to assess genotoxicity of phenanthrene. The strong correlation (r (2) ≥ 0.94) between phenanthrene concentration and DNA damage indicated that comet assay is appropriate for testing the genotoxic effects of phenanthrene-contaminated soil. In the light of these results, we conclude that the Microtox test and comet assay are robust and sensitive bioassays to be employed for the risk evaluation of polycyclic aromatic hydrocarbon-contaminated soil. PMID:23440446

  20. Antimicrobial Activity and Brine Shrimp Lethality Bioassay of the Leaves Extract of Dillenia indica Linn.

    PubMed

    Apu, As; Muhit, Ma; Tareq, Sm; Pathan, Ah; Jamaluddin, Atm; Ahmed, M

    2010-01-01

    The crude methanolic extract of Dillenia indica Linn. (Dilleniaceae) leaves has been investigated for the evaluation of antimicrobial and cytotoxic activities. Organic solvent (n-hexane, carbon tetrachloride and chloroform) fractions of methanolic extract and methanolic fraction (aqueous) were screened for their antimicrobial activity by disc diffusion method. Besides, the fractions were screened for cytotoxic activity using brine shrimp (Artemia salina) lethality bioassay. Among the four fractions tested, n-hexane, carbon tetrachloride, and chloroform fractions showed moderate antibacterial and antifungal activity compared to standard antibiotic, kanamycin. The average zone of inhibition was ranged from 6 to 8 mm at a concentration of 400 µg/disc. But the aqueous fraction was found to be insensitive to microbial growth. Compared to vincristine sulfate (with LC(50) of 0.52 µg/ ml), n-hexane and chloroform fractions demonstrated a significant cytotoxic activity (having LC(50) of 1.94 µg/ml and 2.13 µg/ml, respectively). The LC(50) values of the carbon tetrachloride and aqueous fraction were 4.46 µg/ml and 5.13 µg/ ml, respectively. The study confirms the moderate antimicrobial and potent cytotoxic activities of Dillenia indica leaves extract and therefore demands the isolation of active principles and thorough bioassay. PMID:21331191

  1. Acute and chronic ecotoxicity of carbaryl with a battery of aquatic bioassays.

    PubMed

    Toumi, Hela; Burga-Perez, Karen F; Ferard, Jean-François

    2016-01-01

    The ecotoxic effects of carbaryl (carbamate insecticide) were investigated with a battery of four aquatic bioassays. The nominal effective concentrations immobilizing 50% of Daphnia magna (EC50) after 24 and 48 h were 12.76 and 7.47 µg L(-1), respectively. After 21 days of exposure of D. magna, LOECs (lowest observed effect concentrations) for cumulative molts and the number of neonates per surviving adult were observed at carbaryl concentration of 0.4 µg L(-1). An increase of embryo deformities (curved or unextended shell spines) was observed at 1.8 and 3.7 µg L(-1), revealing that carbaryl could act as an endocrine disruptor in D. magna. Other bioassays of the tested battery were less sensitive: the IC50-72h and IC10-72h of the algae Pseudokirchneriella subcapitata were 5.96 and 2.87 mg L(-1), respectively. The LC50-6d of the ostracod Heterocypris incongruens was 4.84 mg L(-1). A growth inhibition of H. incongruens was registered after carbaryl exposure and the IC20-6d was 1.29 mg L(-1). Our results suggest that the daphnid test sensitivity was better than other used tests. Moreover, carbaryl has harmful and toxic effects on tested species because it acts at low concentrations on diverse life history traits of species and induce embryo deformities in crustaceans. PMID:26549316

  2. Environmental effects of dredging. Environmental effects of dredging technical notes. Plant bioassay of dredged material. Technical notes

    SciTech Connect

    Folsom, B.L.; Lee, C.R.

    1985-06-01

    Recently, a solid-phase plant bioassay was developed to test sediment for contaminants that are potentially phytotoxic and may be bioaccumulated by plants (Folsom and Lee 1981a; Folsom, Lee, and Bates 1981). The solid-phase plant bioassay was shown to be an excellent tool for predicting whether or not contaminants (e.g., zinc and cadmium) were potentially bioaccumulated by the saltwater plant S. alterniflora. Folsom and Lee (1981a) pointed out, however, that the DTPA extraction data indicated that plant uptake from air-dried oxidized saltwater sediment would be substantially greater than from the same saltwater sediment under flooded reduced conditions. In addition, they suspected greater plant uptake once the excess salts were leached out and the sediments were dried. This technical note reports results of modifications to the original solid-phase plant bioassay to pursue this assumption.

  3. Comparison of bioassays for assessing sediment toxicity in Puget Sound. (Appendices included). Report for 1988-1989 (Final)

    SciTech Connect

    Pastorok, R.A.; Becker, D.S.

    1989-05-01

    This study evaluated the relative performance of several bioassays used to assess toxicity of Puget Sound sediments. Selected bioassays were conducted simultaneously on the same array of sediment samples, and direct comparisons were made among the various tests in relation to specific performance criteria. The bioassays and toxicity endpoints evaluated included the following: Amphipod test (Rhepoxynius abronius), percent mortality and abnormality; Amphipod test (Eohaustorius estuarius), percent mortality and abnormality; Juvenile geoduck (Panope generosa), percent mortality; Juvenile polychaete (Neanthes arenaceodentata), percent mortality and percent reduced biomass; Echinoderm embryo test (Dendraster excentricus), percent developmental abnormality and chromosomal abnormality; Bivalve larvae test (Mytilus edulis, Crassostera gigas), percent abnormal development; Microtox saline and organic extract (Photobacterium phosphoreum), percent decrease in luminescence. The 11 evaluation criteria included: dose responsiveness, sensitivity, statistical power, cost-effectiveness, ecological relevance, ease of use, availability of test organisms, endpoint reliability, relationship to indigenous biota, holding constraints, and stage of protocol development.

  4. Use of solid-phase extraction systems to improve the sensitivity of Artemia bioassays for trichothecene mycotoxins.

    PubMed

    Hoke, S H; Carley, C M; Johnson, E T; Broski, F H

    1987-01-01

    Solid-phase extraction was used to preconcentrate trichothecene mycotoxins from rivers and streams in order to develop and improve a rapid and sensitive bioassay using the brine shrimp Artemia salina. For T-2 toxin, HT-2 toxin, and 4,15-diacetoxyscirpenol, LC50 values obtained were 172, 600, and 700 micrograms/L, respectively. The LC50 for 4-deoxynivalenol was 21 mg/L. A more than 5-fold increase in sensitivity was observed when solid-phase extraction (SPE) was used in conjunction with the Artemia bioassay. For T-2 toxin, HT-2 toxin, and 4,15-diacetoxyscirpenol, LC50/SPE values after solid-phase extraction were 21, 83, and 130 micrograms/L. The use of river and stream waters and chlorinated water did not seem to interfere with the bioassay. PMID:3624171

  5. Bioassay of Styrene for Possible Carcinogenicity (CAS No.100-42-5).

    PubMed

    1979-01-01

    Styrene, a widely used intermediate in the manufacture of plastics, elastomers, and resins, was selected for bioassay by the National Cancer Institute because of the widespread use of this compound and a lack of adequate carcinogenicity data. A bioassay for the possible carcinogenicity of styrene was conducted using Fischer 344 rats and B6C3F1 mice. Styrene was administered by gavage to groups of 50 male and 50 female animals of each species. Forty rats of each sex and twenty mice of each sex were placed on test as vehicle controls. The high, medium, and low dosages of styrene administered to rats were, respectively, 2,000, 1,000, and 500 mg/kg. The high and low dosages administered to mice were 300 and 150 mg/kg, respectively. The compound was administered for 78 weeks to high and medium dose rats, for 103 weeks to low dose rats, and for 78 weeks to mice. The period of compound administration was followed by an observation period of 27 weeks for high and medium dose rats, 1 week for low dose rats, and 13 weeks for mice. Mortality among male and female high dose rats was significantly higher than that among their respective vehicle controls. In response to this elevated and early mortality, an additional dosed group of each sex was included in the chronic bioassay. No significant positive association was apparent between dosage and mortality among any other dosed rat groups. For mice, there was a significant positive association between mortality and the dosages of styrene administered to males, but not to females. Adequate numbers of animals in all groups, except for the high dose male and female rats, survived sufficiently long to be at risk from late-developing tumors. Slight dose-related mean body weight depression was apparent when male rats and female mice were compared to their respective vehicle controls, indicating that the dosages administered to these animals during the chronic bioassay may have approximated the maximum tolerated dosages. There was no

  6. A Novel in vitro Bioassay to Explore the Repellent Effects of Compounds Against Mosquito Aedes aegypti (Diptera: Culicidae).

    PubMed

    Rehman, Junaid U; Tabanca, Nurhayat; Khan, Ikhlas A

    2016-01-01

    Mosquitoes are vectors for many pathogens resulting in many deaths of humans. Repellents play an important role in reducing mosquito bites and the spread of mosquito-borne diseases. Currently, Klun & Debboun (K & D) and human-arm-based bioassay systems are used to identify repellent properties of compounds, extracts, and essential oils. Risks involved with human-arm-based systems are allergic reactions and limited replicates. We are reporting an in vitro bioassay method “NCNPR repellent bioassay (NCNPR-RB)” that can closely simulate the results of the cloth patch bioassay system used to determine repellency against mosquitoes. The NCNPRRB method uses heat to attract mosquito and edible collagen sheets as an alternate to human skin. Multiple plant compounds with documented repellency were tested. DEET (N,N-diethyl-3-methylbenzamide) was used as a positive control. Treatments were prepared in EtOH and applied in dosages ranging from 0.011–1.5mg/cm2 to a 20-cm2 collagen sheet. The number of mosquitoes commencing to bite per probe was recorded visually for 1 min. The minimum effective dosage (mg/cm2) of compounds: DEET (0.021), carvacrol (0.011), thymol (0.013), undecanoic acid (0.023), thymol methyl ether (0.269), and 2-nonanone (>0.375 mg/cm2) determined in NCNPRRB were similar to those reported in literature using a cloth patch bioassay system. The NCNPR-RB can be used to screen compounds with reasonable reproducibility of the data at a faster rate than the cloth patch bioassay, which involves the use of human subjects. PMID:26590191

  7. Addressing the recovery of feeding rates in post-exposure feeding bioassays: Cyathura carinata as a case study.

    PubMed

    Pais-Costa, Antonia Juliana; Acevedo, Pelayo; Marques, João Carlos; Martinez-Haro, Mónica

    2015-02-01

    Post-exposure bioassays are used in environmental assessment as a cost-effective tool, but the effects of organism's recovery after exposure to pollutant has not yet been addressed in detail. The recoveries of post-exposure feeding rates after being exposed to two sublethal concentrations of cadmium during two different exposure periods (48h and 96h) were evaluated under laboratory conditions using the estuarine isopod Cyathura carinata. Results showed that feeding depression was a stable endpoint up to 24h after cadmium exposure, which is useful for ecotoxicological bioassays. PMID:25576796

  8. Effects of conidial densities and spray volume of Metarhizium anisopliae and Beauveria bassiana fungal suspensions on conidial viability, droplet size and deposition coverage in bioassay using a novel bioassay spray system

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Experiments were conducted to study the conidial viability during bioassay spray with different suspensions of Metarhizium anisopliae ATCC 62176 and Beauveria bassiana NI8, and to investigate the effects of conidial density and spray volume on the distribution of droplet size and deposit coverage us...

  9. Predicting the carcinogenicity of chemicals in humans from rodent bioassay data.

    PubMed Central

    Goodman, G; Wilson, R

    1991-01-01

    Regulatory agencies currently rely on rodent carcinogenicity bioassay data to predict whether or not a given chemical poses a carcinogenic threat to humans. We argue that it is always more useful to know a chemical's carcinogenic potency (with confidence limits) than to be able to say only qualitatively that it has been found to be a carcinogen. In a typical bioassay, a chemical is administered to groups of 50 to 100 rodents at the highest feasible level (the maximum tolerated dose) and rarely at less than 1/10 this dose in order to maximize the statistical significance of any increase in tumors that might result. Recently, much experimental work has focused on the mechanisms by which site-specific toxicity arising from chronic administration at the maximum tolerated dose may lead to carcinogenicity. Extrapolation of high-dose results to low doses does not take into consideration the possibility of a threshold dose, below which the carcinogenic potency is much lower or even zero. Threshold dose-response phenomena may be much more relevant to the etiology of cancer in the rodent bioassays than was earlier realized; if so, there is an even greater need for establishing dose-dependent potency estimates. The emphasis of this review is on the interspecies comparison of high-dose potencies. The qualitative and quantitative comparison of carcinogenicities between mice and rats and between rodents and humans is reviewed and discussed. We conclude that there is a good qualitative (yes/no) correlation for both the rat/mouse and the rodent/human comparison. There is also a good correlation of the carcinogenic potencies between rats and mice, and the upper limits on potencies in humans are consistent with rodent potencies for those chemicals for which human exposure data are available. For the rodent/human comparison, the best estimate of the interspecies potency factor is lognormally distributed around 1 when the potencies in both species are measured in units of (mg

  10. In vivo genotoxicity of furan in F344 rats at cancer bioassay doses

    SciTech Connect

    Ding, Wei

    2012-06-01

    Furan, a potent rodent liver carcinogen, is found in many cooked food items and thus represents a human cancer risk. Mechanisms for furan carcinogenicity were investigated in male F344 rats using the in vivo Comet and micronucleus assays, combined with analysis of histopathological and gene expression changes. In addition, formamidopyrimidine DNA glycosylase (Fpg) and endonuclease III (EndoIII)-sensitive DNA damage was monitored as a measure of oxidative DNA damage. Rats were treated by gavage on four consecutive days with 2, 4, and 8 mg/kg bw furan, doses that were tumorigenic in 2-year cancer bioassays, and with two higher doses, 12 and 16 mg/kg. Rats were killed 3 h after the last dose, a time established as producing maximum levels of DNA damage in livers of furan-treated rats. Liver Comet assays indicated that both DNA strand breaks and oxidized purines and pyrimidines increased in a near-linear dose-responsive fashion, with statistically significant increases detected at cancer bioassay doses. No DNA damage was detected in bone marrow, a non-target tissue for cancer, and peripheral blood micronucleus assays were negative. Histopathological evaluation of liver from furan-exposed animals produced evidence of inflammation, single-cell necrosis, apoptosis, and cell proliferation. In addition, genes related to apoptosis, cell-cycle checkpoints, and DNA-repair were expressed at a slightly lower level in the furan-treated livers. Although a mixed mode of action involving direct DNA binding cannot be ruled out, the data suggest that furan induces cancer in rat livers mainly through a secondary genotoxic mechanism involving oxidative stress, accompanied by inflammation, cell proliferation, and toxicity. -- Highlights: ► Furan is a potent rodent liver carcinogen and represents a human cancer risk. ► Furan induces DNA damage in rat liver at cancer bioassay doses. ► Furan induces oxidative stress, inflammation and cell proliferation in rat liver. ► Expression of

  11. Theobroma cacao: Review of the Extraction, Isolation, and Bioassay of Its Potential Anti-cancer Compounds.

    PubMed

    Baharum, Zainal; Akim, Abdah Md; Hin, Taufiq Yap Yun; Hamid, Roslida Abdul; Kasran, Rosmin

    2016-02-01

    Plants have been a good source of therapeutic agents for thousands of years; an impressive number of modern drugs used for treating human diseases are derived from natural sources. The Theobroma cacao tree, or cocoa, has recently garnered increasing attention and become the subject of research due to its antioxidant properties, which are related to potential anti-cancer effects. In the past few years, identifying and developing active compounds or extracts from the cocoa bean that might exert anti-cancer effects have become an important area of health- and biomedicine-related research. This review provides an updated overview of T. cacao in terms of its potential anti-cancer compounds and their extraction, in vitro bioassay, purification, and identification. This article also discusses the advantages and disadvantages of the techniques described and reviews the processes for future perspectives of analytical methods from the viewpoint of anti-cancer compound discovery. PMID:27019680

  12. RTD fluxgate performance for application in magnetic label-based bioassay: preliminary results.

    PubMed

    Ando, B; Ascia, A; Baglio, S; Bulsara, A R; Trigona, C; In, V

    2006-01-01

    Magnetic bioassay is becoming of great interest in several application including magnetic separation, drug delivery, hyperthermia treatments, magnetic resonance imaging (MRI) and magnetic labelling. The latter can be used to localize bio-entities (e.g. cancer tissues) by using magnetic markers and high sensitive detectors. To this aim SQUIDs can be adopted, however this result in a quite sophisticated and complex method involving high cost and complex set-up. In this paper, the possibility to adopt RTD fluxgate magnetometers as alternative low cost solution to perform magnetic bio-sensing is investigated. Some experimental results are shown that encourage to pursue this approach in order to obtain simple devices that can detect a certain number of magnetic particles accumulated onto a small surface such to be useful for diagnosis purposes. PMID:17946280

  13. Rapid Bioassay-Guided Isolation of Antibacterial Clerodane Type Diterpenoid from Dodonaea viscosa (L.) Jaeq.

    PubMed Central

    Khurram, Muhammad; Lawton, Linda A.; Edwards, Christine; Iriti, Marcello; Hameed, Abdul; Khan, Murad A.; Khan, Farman A.; ur Rahman, Shafiq

    2015-01-01

    Plant extracts are complex matrices and, although crude extracts are widely in use, purified compounds are pivotal in drug discovery. This study describes the application of automated preparative-HPLC combined with a rapid off-line bacterial bioassay, using reduction of a tetrazolium salt as an indicator of bacterial metabolism. This approach enabled the identification of fractions from Dodonaea viscosa that were active against Staphylococcus aureus and Escherichia coli, which, ultimately, resulted in the identification of a clerodane type diterpenoid, 6β-hydroxy-15,16-epoxy-5β, 8β, 9β, 10α-cleroda-3, 13(16), 14-trien-18-oic acid, showing bacteriostatic activity (minimum inhibitory concentration (MIC) = 64–128 µg/mL) against test bacteria. To the best of our knowledge, this is the first report on antibacterial activity of this metabolite from D. viscosa. PMID:26343638

  14. Methotrexate intercalated layered double hydroxides with the mediation of surfactants: Mechanism exploration and bioassay study.

    PubMed

    Dai, Chao-Fan; Tian, De-Ying; Li, Shu-Ping; Li, Xiao-Dong

    2015-12-01

    Methotrexatum intercalated layered double hydroxides (MTX/LDHs) hybrids were synthesized by the co-precipitation method and three kinds of nonionic surfactants with different hydrocarbon chain lengths were used. The resulting hybrids were then characterized by X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy (TEM). XRD and FTIR investigations manifest the successful intercalation of MTX anions into the interlayer of LDHs. TEM graphs indicate that the morphology of the hybrids changes with the variation of the chain length of the surfactants, i.e., the particles synthesized using polyethylene glycol (PEG-7) present regular disc morphology with good monodispersity, while samples with the mediation of alkyl polyglycoside (APG-14) are heavily aggregated and samples with the addition of polyvinylpyrrolidone (PVP-10) exhibit irregular branches. Furthermore, the release and bioassay experiments show that monodisperse MTX/LDHs present good controlled-release and are more efficient in the suppression of the tumor cells. PMID:26354264

  15. Isolation of bioactive allelochemicals from sunflower (variety Suncross-42) through fractionation-guided bioassays.

    PubMed

    Anjum, Tehmina; Bajwa, Rukhsana

    2010-11-01

    Plants are rich source of biologically active allelochemicals. However, natural product discovery is not an easy task. Many problems encountered during this laborious practice can be overcome through the modification of preliminary trials. Bioassay-directed isolation of active plant compounds can increase efficiency by eliminating many of the problems encountered. This strategy avoids unnecessary compounds, concentrating on potential components and thus reducing the cost and time required. In this study, a crude aqueous extract of sunflower leaves was fractionated through high performance liquid chromatography. The isolated fractions were checked against Chenopodium album and Rumex dentatus. The fraction found active against two selected weeds was re-fractionated, and the active components were checked for their composition. Thin layer chromatography isolated a range of phenolics, whereas the presence of bioactive terpenoids was confirmed through mass spectroscopy and nuclear magnetic resonance spectroscopy. PMID:20981619

  16. Respiratory tract clearance model for dosimetry and bioassay of inhaled radionuclides

    SciTech Connect

    Bailey, M.R.; Birchall, A. ); Cuddihy, R.G. ); James, A.C. ); Roy, M. . Inst. de Protection et de Surete Nucleaire)

    1990-07-01

    The ICRP Task Group on Respiratory Tract Models is developing a model to describe the retention and clearance of deposited radionuclides for dose-intake calculations and interpretation of bioassay data. Clearance from each region is treated as competition between mechanical transport, which moves particles to the gastro-intestinal tract and lymph nodes, and the translocation of material to blood. It is assumed that mechanical transport rates are the same for all materials, and that rates of translocation to blood are the same in all regions. Time-dependent clearance is represented by combinations of compartments. Representative values of parameters to describe mechanical transport from the human respiratory tract have been estimated, and guidance is given on the determination of translocation rates. It is emphasized that the current version of the model described here is still provisional. 30 refs.

  17. Monitoring North Sea oil production discharges using passive sampling devices coupled with in vitro bioassay techniques.

    PubMed

    Harman, Christopher; Farmen, Eivind; Tollefsen, Knut Erik

    2010-09-01

    Semipermeable membrane devices (SPMDs) and polar organic integrative chemical samplers (POCIS) were deployed in vicinity of an offshore oil production platform discharging production water (produced water) to the North Sea. Extracts from SPMDs and POCIS were subjected to chemical analysis for polycyclic aromatic hydrocarbons (PAHs) and alkylphenols (APs) respectively, and also assessed for acute toxicity (cytotoxicity), estrogen receptor (ER)-mediated production of vitellogenin (Vtg) and induction of 7-ethoxyresorufin-O-deethylase (EROD) activity in primary hepatocytes from rainbow trout (Oncorhynchus mykiss). Chemical analysis of the extracts revealed a gradient of exposure away from the platform for low molecular weight PAH and AP, whereas no exposure gradient was apparent for high molecular weight PAH, as expected. These data coupled with earlier work allowed a tentative general exposure scenario to be determined. The passive sampler extracts also caused modulation of the bioassay toxicity endpoints, although a clear gradient of response relative to the discharge point could not be identified. PMID:20683536

  18. Preserved echinoderm gametes as a useful and ready-to-use bioassay material.

    PubMed

    Kiyomoto, M; Hamanaka, G; Hirose, M; Yamaguchi, M

    2014-02-01

    Marine animals, and sea urchin species in particular, have several advantages for use in environmental research. However, the spawned eggs of the sea urchin quickly lose fertility, although the fertile period can be lengthened by the addition of antibiotics to the sea water (Epel et al., 2004). We evaluated five species of Japanese sea urchin and the gametes of Hemicentrotus pulcherrimus could be maintained for 2 weeks or more at low temperature with the addition of antibiotics to sea water. We also demonstrated the practicality of shipping these preserved gametes as experimental material for universities and schools to use immediately for bioassays of physical and chemical impacts on the marine environment. PMID:24129269

  19. Correlation and comparison of Nb/sub 2/ lymphoma cell bioassay with radioimmunoassay for human prolactin

    SciTech Connect

    Subramanian, M.G.; Spirtos, N.J.; Moghissi, K.S.; Magyar, D.M.; Hayes, M.F.; Gala, R.R.

    1984-12-01

    Serum samples from groups of men and women with normal and elevated prolactin (PRL) levels were assayed by radioimmunoassay (RIA) and by Nb/sub 2/ lymphoma cell bioassay (BA) for the presence of PRL. Because the Nb/sub 2/ lymphoma cells respond to both PRL and growth hormone, BA for PRL activity was carried out before and after neutralization of growth hormone in the serum samples. There were excellent correlations between RIA and BA both in euprolactinemic and hyperprolactinemic subjects. On an absolute basis, RIA and BA values were similar in the euprolactinemic group (6.6 +/- 0.8 versus 6.2 +/- 1.0), whereas in the hyperprolactinemic group, RIA values were significantly higher than the BA results. The two assay systems also appeared to correlate better in women who were hyperprolactinemic, with obvious menstrual cycle disturbances, than in hyperprolactinemic women without menstrual cycle disturbances.

  20. Evaluation of the toxic and genotoxic potential of acid mine drainage using physicochemical parameters and bioassays.

    PubMed

    Netto, E; Madeira, R A; Silveira, F Z; Fiori, M A; Angioleto, E; Pich, C T; Geremias, R

    2013-05-01

    Carboniferous activity generates acid mine drainage (AMD) which is capable of unleashing toxic effects on the exposed biota. The aim of this study was to evaluate the toxic and genotoxic potential of untreated-AMD and AMD treated with calcinated sediment, using physicochemical parameters and bioassays. Results revealed that untreated-AMD presented low pH values and elevated concentrations of the metals Fe, Al, Mn, Zn and Cu. High acute toxicity was observed in Artemia sp. and Daphnia magna, and sub-chronic toxicity and genotoxicity in Allium cepa L. as well as scission of plasmid DNA exposed to untreated-AMD. Treatment of AMD with calcinated sediment promoted the reduction of acidity and the removal of metals, as well as a reduction in toxic and genotoxic effects. In conclusion, the calcinated sediment can be used as an alternative AMD treatment. PMID:23518284

  1. Bioassay technique using nonspecific esterase activities of Tetrahymena pyriformis for screening and assessing cytotoxicity of xenobiotics

    SciTech Connect

    Bogaerts, P.; Senaud, J.; Bohatier, J. |

    1998-08-01

    A simple and rapid test for screening and assessing the cytotoxicity of xenobiotics was developed with Tetrahymena pyriformis. The method estimates the activities of nonspecific esterases of a cell by concentrating within it a specific amount of fluorescence associated with fluorescein dye. The 2-h median effective concentration (EC50) values of 10 inorganic and eight organic substances are presented and compared to those of three other bioassays: the conventional T. pyriformis proliferation rate 9-h median inhibitory concentrations, the Microtox 30-min EC50s, and the Daphnia magna 4-methylumbelliferyl {beta}-D galactoside 1-h EC50s. A highly significant correlation was found between the results obtained with the fluorescein diacetate test and those obtained with the growth inhibition and Microtox tests. This in vivo enzymatic test showed high sensitivity to all compounds tested except Cr{sup 6+} and sodium dodecyl sulfate.

  2. Laboratory bioassays of entomopathogenic fungi for control of Delia radicum (L.) larvae.

    PubMed

    Bruck, Denny J; Snelling, Jane E; Dreves, Amy J; Jaronski, Stefan T

    2005-06-01

    Laboratory soil bioassays were performed at economic field rates for in-furrow (3.85 x 10(6)spores/g dry soil) and broadcast (3.85 x 10(5)spores/g dry soil) applications with three isolates of Metarhizium anisopliae (F52, ATCC62176, and ARSEF5520) and one isolate of Beauveria bassiana (GHA). All isolates tested were infective to second instar Delia radicum (L.). The conditionally registered M. anisopliae isolate (F52) performed best killing an average of 85 and 72% of D. radicum larvae at the high and low concentration, respectively. The mean LC50 and LC95 of F52 against second instar D. radicum was 2.7 x 10(6) and 1.8 x 10(8)spores/g dry soil, respectively. The use of F52 in an integrated management program is discussed. PMID:16087004

  3. Harvester ant bioassay for assessing hazardous chemical waste sites. [Pogonomyrmex owhyeei

    SciTech Connect

    Gano, K.A.; Carlile, D.W.; Rogers, L.E.

    1985-05-01

    A technique was developed for using harvester ants, Pogonomyrmex owhyeei, in terrestrial bioassays. Procedures were developed for maintaining stock populations, handling ants, and exposing ants to toxic materials. Relative toxicities were determined by exposing ants to 10 different materials. These materials included three insecticides, Endrin, Aldrin, and Dieldrin; one herbicide, 2,4-D; three complex industrial waste residuals, wood preservative sludge, drilling fluid, and slop oil; and three heavy metals, copper zinc, and cadium. Ants were exposed in petri dishes containing soil amended with a particular toxicant. Under these test conditions, ants showed no sensitivity to the metals or 2,4-D. Ants were sensitive to the insecticides and oils in repeated tests, and relative toxicity remained consistent throughout. Aldrin was the most toxic material followed by Dieldrin, Endrin, wood preservative sludge, drilling fluid, and slop oil. 12 refs., 2 figs., 2 tabs.

  4. Environmental effects of dredging: Upland animal bioassays of dredged materials. Technical note

    SciTech Connect

    Simmers, J.W.; Rhett, R.G.; Lee, C.R.

    1986-01-01

    The Clean Water Act in the United States requires that the environmental evaluation of dredged material prior to discharge or impacting the waters of the United States include the effects of disposal on concentrations of contaminants through biological processes. This results in a need for Corps of Engineers districts to be able to predict the contamination of animals that may be associated with potential disposal alternatives: open-water disposal, upland disposal, and wetland creation. The following is a summary of the results of bioassay procedures using the earthworm Eisenia foetida to evaluate the potential contaminant mobility into soil-dwelling animals. These tests were derived from proposed Organization for European Common Development (OECD) and European Economics Commission (EEC) test procedures (evaluating the effects of new chemicals) and modified to consider accumulation and sublethal effects rather than toxicity.

  5. Theobroma cacao: Review of the Extraction, Isolation, and Bioassay of Its Potential Anti-cancer Compounds

    PubMed Central

    Baharum, Zainal; Akim, Abdah Md; Hin, Taufiq Yap Yun; Hamid, Roslida Abdul; Kasran, Rosmin

    2016-01-01

    Plants have been a good source of therapeutic agents for thousands of years; an impressive number of modern drugs used for treating human diseases are derived from natural sources. The Theobroma cacao tree, or cocoa, has recently garnered increasing attention and become the subject of research due to its antioxidant properties, which are related to potential anti-cancer effects. In the past few years, identifying and developing active compounds or extracts from the cocoa bean that might exert anti-cancer effects have become an important area of health- and biomedicine-related research. This review provides an updated overview of T. cacao in terms of its potential anti-cancer compounds and their extraction, in vitro bioassay, purification, and identification. This article also discusses the advantages and disadvantages of the techniques described and reviews the processes for future perspectives of analytical methods from the viewpoint of anti-cancer compound discovery. PMID:27019680

  6. Rotationally Induced Hydrodynamics: Fundamentals and Applications to High-Speed Bioassays

    NASA Astrophysics Data System (ADS)

    Wang, Gufeng; Driskell, Jeremy D.; Hill, April A.; Dufek, Eric J.; Lipert, Robert J.; Porter, Marc D.

    2010-07-01

    Bioassays are indispensable tools in areas ranging from fundamental life science research to clinical practice. Improving assay speed and levels of detection will have a profound impact in all of these areas. We recently developed a rapid, sensitive format for immunosorbent assays that expedites antigen mass transport by rotating the capture substrate. This review outlines the theoretical foundation of rotationally induced hydrodynamics and its application in heterogeneous assays. We describe a general solution that solves the rates of immunoreactions on rotating capture substrates, taking into account both diffusion and the rate of reaction between antibody and antigen. The general solution applies to a wide range of rotation rates, including mass transport-limited to reaction rate-limited assays, and is validated experimentally. We discuss several applications that demonstrate how immunoassays can be tailored to increase speed as well as lower the limit of detection of viral particles, pathogens, toxins, and proteins.

  7. Ecotoxicological assessment of metal-polluted urban soils using bioassays with three soil invertebrates.

    PubMed

    Santorufo, Lucia; Van Gestel, Cornelis A M; Maisto, Giulia

    2012-07-01

    This study aimed at assessing the quality of urban soils by integrating chemical and ecotoxicological approaches. Soils from five sites in downtown Naples, Italy, were sampled and characterized for physical-chemical properties and total and water-extractable metal concentrations. Bioassays with Eisenia andrei, Enchytraeus crypticus and Folsomia candida were performed to assess toxicity of the soils, using survival, reproduction and growth as the endpoints. Metal bioaccumulation in the animals was also measured. The properties and metal concentrations of the soils strongly differed. Metal bioaccumulation was related with total metal concentrations in soil and was highest in E. crypticus, which was more sensitive than E. andrei and F. candida. Responses of the three species to the investigated soils seemed due to both metal contamination and soil properties. PMID:22445389

  8. Bioassay for assessing cell stress in the vicinity of radio-frequency irradiating antennas.

    PubMed

    Monselise, Edna Ben-Izhak; Levkovitz, Aliza; Gottlieb, Hugo E; Kost, Daniel

    2011-07-01

    The 24 h exposure of water plants (etiolated duckweed) to RF-EMF between 7.8 V m(-1) and 1.8 V m(-1), generated by AM 1.287 MHz transmitting antennas, resulted in alanine accumulation in the plant cells, a phenomenon we have previously shown to be a universal stress signal. The magnitude of the effect corresponds qualitatively to the level of RF-EMF exposure. In the presence of 10 mM vitamin C, alanine accumulation is completely suppressed, suggesting the involvement of free radicals in the process. A unique biological connection has thus been made between exposure to RF-EMF and cell stress, in the vicinity of RF transmitting antennas. This simple test, which lasts only 24 h, constitutes a useful bioassay for the quick detection of biological cell stress caused in the vicinity of RF irradiating antennas. PMID:21655615

  9. Submicron polyacrolein particles in situ embedded with upconversion nanoparticles for bioassay.

    PubMed

    Generalova, A N; Kochneva, I K; Khaydukov, E V; Semchishen, V A; Guller, A E; Nechaev, A V; Shekhter, A B; Zubov, V P; Zvyagin, A V; Deyev, S M

    2015-02-01

    We report a new surface modification approach of upconversion nanoparticles (UCNPs) structured as inorganic hosts NaYF4 codoped with Yb(3+) and Er(3+) based on their encapsulation in a two-stage process of precipitation polymerization of acrolein under alkaline conditions in the presence of UCNPs. The use of tetramethylammonium hydroxide both as an initiator of acrolein polymerization and as an agent for UCNP hydrophilization made it possible to increase the polyacrolein yield up to 90%. This approach enabled the facile, lossless embedment of UCNPs into the polymer particles suitable for bioassay. These particles are readily dispersible in aqueous and physiological buffers, exhibiting excellent photoluminescence properties, chemical stability, and also allow the control of particle diameters. The feasibility of the as-produced photoluminescent polymer particles mean-sized 260 nm for in vivo optical whole-animal imaging was also demonstrated using a home-built epi-luminescence imaging system. PMID:25510961

  10. Rapid, Bioassay-Guided Process for the Detection and Identification of Antibacterial Neem Oil Compounds.

    PubMed

    Krüzselyi, Dániel; Nagy, Róbert; Ott, Péter G; Móricz, Ágnes M

    2016-08-01

    Bioassay guidance was used along the whole process including method development, isolation and identification of antibacterial neem (Azadirachta indica) oil compounds. The biomonitoring was performed by direct bioautography (DB), a combination of thin-layer chromatography (TLC) and antimicrobial detection. DB of neem oil showed one antibacterial zone that was not UV-active; therefore, the TLC separation was improved under DB control. The chromatographic zone that exhibited activity against Bacillus subtilis, Xanthomonas euvesicatoria, Aliivibrio fischeri, Staphylococcus aureus and methicillin-resistant Staphylococcus aureus was characterized by TLC reagents, indicating a lipophilic, fatty acid-like chemical feature. Two compounds were found and identified in the active zone by high-performance liquid chromatography-electrospray ionization mass spectrometry as linoleic and oleic acids. Both fatty acids inhibited B. subtilis, but A. fischeri was sensitive only against linoleic acid. PMID:26951543

  11. Bioassay Guided Fractionation Identified Hederagenin as a Major Cytotoxic Agent from Cyclocarya paliurus Leaves.

    PubMed

    Gao, Ying; He, Chunnian; Bi, Wu; Wu, Guofan; Altman, Elliot

    2016-01-01

    An ethanol extract prepared from the leaves of Cyclocarya paliurus, also known as sweet tea, which is one of the most popular teas utilized in traditional Chinese medicine, exhibited significant cytotoxicity against human lung and breast cancer cells. Using a bioassay-guided fractionation, we purified a pentacyclic triterpenoid, hederagenin, which exhibited superior and selective cytotoxicity against human breast and lung cancer cells. Evaluation of the structure-activity relationship between hederagenin and seven other pentacyclic triterpenoids revealed that the C3 hydroxyl group, the C17 carboxyl group and the Δ (12,13) double bond could be important active groups for the bioactivity of pentacyclic triterpenoids, whereas introduction of a hydroxyl group at C2 or C23 might reduce their bioactivity. We also investigated the cytotoxic activity of hedeargenin and demonstrated that it induces apoptosis, increases the cell membrane permeability, reduces the mitochondria potential, and suppresses NF-κB activation. PMID:26393939

  12. Bioassay-guided isolation of prenylated xanthones and polycyclic acylphloroglucinols from the leaves of Garcinia nujiangensis.

    PubMed

    Xia, Zheng-Xiang; Zhang, Dan-Dan; Liang, Shuang; Lao, Yuan-Zhi; Zhang, Hong; Tan, Hong-Sheng; Chen, Shi-Lin; Wang, Xin-Hong; Xu, Hong-Xi

    2012-08-24

    Bioassay-guided fractionation of the acetone extract of the leaves of Garcinia nujiangensis resulted in the isolation of two new prenylated xanthones, nujiangexanthones A (1) and B (2), three new polycyclic polyprenylated acylphloroglucinols, nujiangefolins A-C (3-5), and 10 known related analogues. The structures of compounds 1-5 were elucidated by interpretation of their spectroscopic data. Compounds 3 and 4 are unusual polycyclic polyprenylated acylphloroglucinols in which the enol hydroxy group forms a six-membered ring with a benzene ring carbon. The compounds isolated were evaluated for their cytotoxic effects against 11 cancer cell lines and immortalized MIHA normal liver cells, and the test substances demonstrated selectivity toward the cancer cells. Isojacareubin (6) was found to be the most potent cytotoxic compound of those tested. PMID:22871217

  13. Bioassay-guided isolation of xanthones and polycyclic prenylated acylphloroglucinols from Garcinia oblongifolia.

    PubMed

    Huang, Sheng-Xiong; Feng, Chao; Zhou, Yan; Xu, Gang; Han, Quan-Bin; Qiao, Chun-Feng; Chang, Donald C; Luo, Kathy Qian; Xu, Hong-Xi

    2009-01-01

    Bioassay-guided fractionation of the acetone extract of the bark of Garcinia oblongifolia has resulted in the isolation of three new xanthones, oblongixanthones A-C (1-3), three new polyprenylated benzoylphloroglucinols, oblongifolins E-G (4-6), and 12 known compounds. Oblongifolins I (5) and J (6) are the first natural products that have similar structural features to those of two known oxidation products of garcinol. The structures of the new compounds 1-6 were characterized by spectroscopic data interpretation. All isolates were assayed for their apoptosis-inducing effects against HeLa-C3 cells. Oblongifolin C (16) was found to be the most potent apoptotic inducer of the compounds evaluated. PMID:19113969

  14. Bio-assay Guided Isolation of Anti-cancer Compounds from Anthocephalus cadamba Bark.

    PubMed

    Kumar, Deepak; Tejaswi, Chilukuri; Rasamalla, Saiprasanna; Mallick, Sumana; Pala, Bikas C

    2015-08-01

    Anthocephalus cadamba, an important plant in the traditional system of medicine in India, is reported to possess anticancer activity. Guided by bio-assay tests using human colorectal (HCT116) and hepatocellular carcinoma (HepG2) cell lines, it has been shown to contain three active constituents, the triterpenoid saponins 3-O-[α-L-rhamnopyranosyl]-quinovic acid (1) and 3-O-[α-L-rhamnopyranosyl]-quinovic acid 28-O-[β-D-glucopyranosyl] ester (2), and the alkaloid cadambine (3). The structures of the isolated compounds were established using spectroscopic techniques. The isolated compounds demonstrated concentration dependent inhibition of both the cell lines, where compound 3 proved to be the most potent inhibitor of cell line HCT116 (IC50 45 +/- 4 μg/mL) and compound 2 demonstrated maximum inhibitory activity against HepG2 cell line with an IC50 value of 89 +/- 7 μg/mL. PMID:26434112

  15. Turbidity as a method of preparing sperm dilutions in the echinoid sperm/egg bioassay

    SciTech Connect

    Hall, T.J.; Haley, R.K.; Battan, K.J. )

    1993-11-01

    The use of turbidimeter for preparing sperm dilutions used in the echinoid sperm/egg bioassay was evaluated. Regression analyses of the relationship between sperm density and turbidity for the sea urchins Strongylocentrotus purpuratus and Strongylocentrotus droebachiensis and the sand dollar Dendraster excentricus indicated that although there were slope differences for each species, each coefficient of determination was highly significant. For Dendraster excentricus, triplicate hemacytometer counts over a range of turbidities as well as repeated preparations of a single sperm turbidity indicated similar variability for each. The use of the turbidimeter has time-saving advantages over conventional hemacytometer methods without sacrificing precision. Sperm dilutions can be prepared rapidly, minimizing seawater sperm preactivation before test initiation, and may therefore contribute to increased test precision.

  16. Using macroalgal δ15N bioassay to detect cruise ship waste water effluent inputs in Skagway, AK

    EPA Science Inventory

    Nitrogen stable isotopes are a powerful tool for tracking sources of N to marine ecosystems. I used green macroalgae as a bioassay organism to evaluate if the δ15N signature of cruise ship waste water effluent (CSWWE) could be detected in Skagway Harbor, AK. Opportunistic green...

  17. USE OF BIOASSAY-DIRECTED CHEMICAL ANALYSIS FOR IDENTIFYING MUTAGENIC COMPOUNDS IN URBAN AIR AND COMBUSTION EMISSIONS

    EPA Science Inventory

    Bioassay-directed chemical analysis fractionation has been used for 30 years to identify mutagenic classes of compounds in complex mixtures. Most studies have used the Salmonella (Ames) mutagenicity assay, and we have recently applied this methodology to two standard reference sa...

  18. USE OF PLANT AND EARTHWORM BIOASSAYS TO EVALUATE REMEDIATION OF SOIL FROM A SITE CONTAMINATED WITH POLYCHLORINATED BIPHENYLS

    EPA Science Inventory

    Soil from a site heavily contaminated with polychlorinated biphenyls (PCBs) was treated with a pilot-scale, solvent extraction technology. Bioassays in earthworms and plants were used to examine the efficacy of the remediation process for reducing the toxicity of the soil. The ...

  19. A fish-feeding laboratory bioassay to assess the antipredatory activity of secondary metabolites from the tissues of marine organisms.

    PubMed

    Marty, Micah J; Pawlik, Joseph R

    2015-01-01

    Marine chemical ecology is a young discipline, having emerged from the collaboration of natural products chemists and marine ecologists in the 1980s with the goal of examining the ecological functions of secondary metabolites from the tissues of marine organisms. The result has been a progression of protocols that have increasingly refined the ecological relevance of the experimental approach. Here we present the most up-to-date version of a fish-feeding laboratory bioassay that enables investigators to assess the antipredatory activity of secondary metabolites from the tissues of marine organisms. Organic metabolites of all polarities are exhaustively extracted from the tissue of the target organism and reconstituted at natural concentrations in a nutritionally appropriate food matrix. Experimental food pellets are presented to a generalist predator in laboratory feeding assays to assess the antipredatory activity of the extract. The procedure described herein uses the bluehead, Thalassoma bifasciatum, to test the palatability of Caribbean marine invertebrates; however, the design may be readily adapted to other systems. Results obtained using this laboratory assay are an important prelude to field experiments that rely on the feeding responses of a full complement of potential predators. Additionally, this bioassay can be used to direct the isolation of feeding-deterrent metabolites through bioassay-guided fractionation. This feeding bioassay has advanced our understanding of the factors that control the distribution and abundance of marine invertebrates on Caribbean coral reefs and may inform investigations in diverse fields of inquiry, including pharmacology, biotechnology, and evolutionary ecology. PMID:25650625

  20. 2,3,7,8-TCDD toxic equivalents in Great Lakes salmon derived using mammalian, bird, and fish cell bioassays

    SciTech Connect

    Heuvel, M.R. van den; Clemons, J.H.; Bols, N.C.; Dixon, D.G.; Kennedy, S.W.; Metcalfe, T.L.; Smith, I.R.

    1994-12-31

    Chinook salmon from lakes Ontario and Huron, and Coho salmon from lakes Ontario and Erie were captured during their fall spawning migration. Subsamples of extracted pooled muscle, liver and egg tissue were used to measure congener specific PCBs, chlorinated dioxins and furans as well as bioassay derived 2,3,7,8-TCDD toxic equivalent concentration (TEC). The cell culture bioassays used to measure TECs were rat hepatoma (H411E) and rainbow trout hepatocyte (RTL-W1) continuous cell lines as well as chicken embryo hepatocyte primary culture (CEH). Although excellent correlations were found between all 3 cell culture bioassays, CEH was found to be 10 times and 30 times more sensitive than H411E and RTL-W1 respectively. Lake Ontario TECs were found to be higher than either Lake Huron or Lake Erie samples for both species of Salmon, and Chinook salmon was elevated over Coho Salmon. Chemical data indicates that the more toxic coplanar PCBs are selectively concentrated in eggs as compared to liver and muscle. Bioassay derived TECs are discussed with regard to chemical contribution of the PCB and dioxin/furan congeners based on an additive model.

  1. Elemol and Amyris Oil Repel the Ticks Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae) in Laboratory Bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The essential oil from Amyris balsamifera (Rutaceae) and elemol, a principal constituent of the essential oil of Osage orange, Maclura pomifera (Moraceae) were evaluated in in vitro and in vivo laboratory bioassays for repellent activity against host-seeking nymphs of the blacklegged tick, Ixodes sc...

  2. Cumulative toxicity of an environmentally relevant mixture of nine regulated disinfection by-products in a multigenerational rat reproductive bioassay

    EPA Science Inventory

    CUMULATIVE TOXICITY OF AN ENVIRONMENTALLY RELEVANT MIXTURE OF NINE REGULATED DISINFECTION BY-PRODUCTS IN A MULTIGENERATIONAL RAT REPRODUCTIVE BIOASSAY J E Simmons, GR. Klinefelter, JM Goldman, AB DeAngelo, DS Best, A McDonald, LF Strader, AS Murr, JD Suarez, MH George, ES Hunte...

  3. Evaluation of the toxic effects of arsenite, chromate, cadmium, and copper using a battery of four bioassays.

    PubMed

    Ko, Kyung-Seok; Lee, Pyeong-Koo; Kong, In Chul

    2012-09-01

    The sensitivities of four different kinds of bioassays to the toxicities of arsenite, chromate, cadmium, and copper were compared. The different bioassays exhibited different sensitivities, i.e., they responded to different levels of toxicity of each of the different metals. However, with the exception of the α-glucosidase enzyme activity, arsenite was the most toxic compound towards all the tested organisms, exhibiting the highest toxic effect on the seeds of Lactuca, with an EC(50) value of 0.63 mg/L. The sensitivities of Lactuca and Raphanus were greater than the sensitivities of two other kinds of seeds tested. Therefore, these were the seeds appropriate for use in a seed germination assay. A high revertant mutagenic ratio (5:1) of Salmonella typhimurium was observed with an arsenite concentration of 0.1 μg/plate, indicative of a high possibility of mutagenicity. These different results suggested that a battery of bioassays, rather than one bioassay alone, is needed as a more accurate and better tool for the bioassessment of environmental pollutants. PMID:22170103

  4. Syringe test (modified larval immersion test): a new bioassay for testing acaricidal activity of plant extracts against Rhipicephalus microplus.

    PubMed

    Sindhu, Zia-ud-Din; Jonsson, Nicholas N; Iqbal, Zafar

    2012-09-10

    We report a new bioassay "syringe test" (modified larval immersion test) for in vitro evaluation of acaricidal activity of crude plant extracts. Prepared syringes, containing eggs of tick, were incubated until 14 d after hatching of eggs, when the bioassay was performed on the larvae. Lethal concentrations for 50% of larvae (LC(50)), LC(90) and LC(99) values were calculated for each tested product. 95% confidence intervals for LC(50) were very narrow, indicating a high degree of repeatability for the new bioassay on larvae of R. microplus. Bioassays were applied to six crude aqueous-methanol extracts from five plants (Acacia nilotica, Buxus papillosa, Fumaria parviflora, Juniperus excelsa, and Operculina turpethum), of which three showed discernible effects. Twenty-four hours post exposure, LC(99) values were 11.9% (w/v) for F. parviflora, 20.8% (w/v) and 29.2% (w/v) for B. papillosa and A. nilotica, respectively. After six days of exposure these values were; 9.1% (w/v), 9.2% (w/v) and 15.5 (w/v) for F. parviflora, A. nilotica and B. papillosa, respectively. PMID:22516644

  5. Proof of concept for a novel insecticide bioassay based on sugar feeding by adult Aedes aegypti (Stegomyia aegypti).

    PubMed

    Stell, F M; Roe, R M; Arellano, C; Kennedy, L; Thornton, H; Saavedra-Rodriguez, K; Wesson, D M; Black, W C; Apperson, C S

    2013-09-01

    Aedes aegypti L. (Stegomyia aegypti) (Diptera: Culicidae) is the principal vector of dengue and yellow fever viruses in tropical and subtropical regions of the world. Disease management is largely based on mosquito control achieved by insecticides applied to interior resting surfaces and through space sprays. Population monitoring to detect insecticide resistance is a significant component of integrated disease management programmes. We developed a bioassay method for assessing insecticide susceptibility based on the feeding activity of mosquitoes on plant sugars. Our prototype sugar-insecticide feeding bioassay system was composed of inexpensive, disposable components, contained minimal volumes of insecticide, and was compact and highly transportable. Individual mosquitoes were assayed in a plastic cup that contained a sucrose-permethrin solution. Trypan blue dye was added to create a visual marker in the mosquito's abdomen for ingested sucrose-permethrin solution. Blue faecal spots provided further evidence of solution ingestion. With the sugar-insecticide feeding bioassay, the permethrin susceptibility of Ae. aegypti females from two field-collected strains was characterized by probit analysis of dosage-response data. The field strains were also tested by forced contact of females with permethrin residues on filter paper. Dosage-response patterns were similar, indicating that the sugar-insecticide feeding bioassay had appropriately characterized the permethrin susceptibility of the two strains. PMID:23077986

  6. DUCKWEED (LEMNA GIBBA) GROWTH INHIBITION BIOASSAYS FOR EVALUATING THE TOXICITY OF OLIVE MILL WASTES BEFORE AND DURING COMPOSTING

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two-phase olive mill waste (TPOMW) is considered a major problem confronting the modern oil extraction and processing industry. Composting has been recently proposed as a suitable method to treat TPOMW so that it is suitable for use in agriculture. In the work reported here, the Lemna gibba bioassay...

  7. COMPARATIVE ESTROGENICITY OF ESTRADIOL, ETHYNYL ESTRADIOL AND DIETHYLSTILBESTROL IN AN IN VIVO, MALE SHEEPSHEAD MINNOW (CYPRINODON VARIEGATUS), VITELLOGENIN BIOASSAY

    EPA Science Inventory

    An in vivo bioassay for vitellogenin (VTG) synthesis was developed to screen individual chemicals or mixtures of chemicals for potentially estrogenic effects in a marine teleost model. An enzyme-linked immunosorbent assay (ELISA) was used to quantitate VTG synthesis in male sheep...

  8. IDENTIFICATION OF ANTIFUNGAL COMPOUNDS IN A BIOLOGICAL CONTROL PRODUCT USING A MICROPLATE INHIBITION BIOASSAY AND MS ANALYSIS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An inhibition assay was developed to quantify the antifungal activity of a solid state fermentation of Bacillus subtilis. Methanol extracts were tested against a spore solution of the fungal pathogen Fusarium oxysporum f. sp. melonis using a 96-well microplate bioassay. HPLC analysis of extracts s...

  9. Cleavable DNA-protein hybrid molecular beacon: A novel efficient signal translator for sensitive fluorescence anisotropy bioassay.

    PubMed

    Hu, Pan; Yang, Bin

    2016-01-15

    Due to its unique features such as high sensitivity, homogeneous format, and independence on fluorescent intensity, fluorescence anisotropy (FA) assay has become a hotspot of study in oligonucleotide-based bioassays. However, until now most FA probes require carefully customized structure designs, and thus are neither generalizable for different sensing systems nor effective to obtain sufficient signal response. To address this issue, a cleavable DNA-protein hybrid molecular beacon was successfully engineered for signal amplified FA bioassay, via combining the unique stable structure of molecular beacon and the large molecular mass of streptavidin. Compared with single DNA strand probe or conventional molecular beacon, the DNA-protein hybrid molecular beacon exhibited a much higher FA value, which was potential to obtain high signal-background ratio in sensing process. As proof-of-principle, this novel DNA-protein hybrid molecular beacon was further applied for FA bioassay using DNAzyme-Pb(2+) as a model sensing system. This FA assay approach could selectively detect as low as 0.5nM Pb(2+) in buffer solution, and also be successful for real samples analysis with good recovery values. Compatible with most of oligonucleotide probes' designs and enzyme-based signal amplification strategies, the molecular beacon can serve as a novel signal translator to expand the application prospect of FA technology in various bioassays. PMID:26592607

  10. MORTALITY OF TARNISHED PLANT BUG TO VARIOUS INSECTICIDES IN SPRAY TABLE BIOASSAYS FOLLOWING FIELD APPLICATION OF CURACRON, 2001.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The efficacy of various insecticides on mortality of the tarnished plant bug collected at 48h following a field application of Curacron at 0.50 lb (AI)/acre were evaluated in laboratory bioassays. Treatments were Orthene, Curacron, Bidrin, Regent, Steward, and Baythroid applied to cotton terminals ...

  11. Identification of Androgen Receptor Antagonists in Fish Using a Simple Bioassay with the Fathead Minnow Pimephales promelas .

    EPA Science Inventory

    Considerable effort has been expended on the development of bioassays to detect chemicals that affect endocrine function controlled by the vertebrate hypothalamic-pituitary-gonadal (HPG) axis via different mechanisms/modes of action (MOA). Antagonism of the androgen receptor (AR)...

  12. Development and testing of a laboratory spray table methodology to bioassay simulated levels of aerial spray drift

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this work was to develop a repeatable methodology for bioassaying simulated levels of aerially applied glyphosate at deposition levels ranging from 1/3 to 1/100 of labeled rate at droplet sizes of 100 µm in a spray table environment. These drift deposition levels are consistent wit...

  13. MORTALITY OF TARNISHED PLANT BUGS TO VARIOUS INSECTICIDES IN SPRAY TABLE BIOASSAYS FOLLOWING FIELD APPLICATION OF ORTHENE, 2001

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The efficacy of various insecticides on mortality of the tarnished plant bug collected at 48h following a field application of Orthene at 0.50 lb (AI)/acre were evaluated in laboratory bioassays. Treatments were Vydate, Orthene, Regent, Centric, Provado, and Decis applied to cotton terminals at 0.3...

  14. 9 CFR 147.16 - Procedure for the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment).

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Procedure for the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment). 147.16 Section 147.16 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE LIVESTOCK IMPROVEMENT...

  15. Quantification of follicle stimulating hormone (follitropin alfa): is in vivo bioassay still relevant in the recombinant age?

    PubMed

    Driebergen, R; Baer, G

    2003-01-01

    Quantification of follicle-stimulating hormone (FSH) for clinical use has traditionally involved the use of in vivo bioassays, particularly the Steelman-Pohley bioassay. This assay has limited precision, requires large numbers of laboratory animals and involves cumbersome procedures for data generation and interpretation. Recent advances in manufacturing procedures for recombinant human FSH (r-hFSH) have resulted in a preparation (follitropin alfa; Gonal-F) that is highly consistent in both isoform profile and glycan species distribution. As a result, follitropin alfa can be reliably quantified using an optimised size exclusion high-performance liquid chromatography (SE-HPLC) method, and vials can be filled by mass. Preliminary clinical studies suggest that the fill-by-mass process results in a product that delivers a more consistent clinical response and is more effective than follitropin alfa vials filled by bioassay in women undergoing controlled ovarian stimulation. Non-bioassay methods such as SE-HPLC are likely to become increasingly important for quality testing and regulatory purposes, provided that the manufacturing process is well controlled and produces a protein of highly consistent physico-chemical properties. PMID:12661779

  16. Effects of sporophyll storage on giant kelp Macrocystis pyrifera (Agardh) bioassay

    SciTech Connect

    Gully, J.R.; Bottomley, J.P.; Baird, R.B.

    1999-07-01

    The giant kelp Macrocystis pyrifera (Agardh) is a US Environmental Protection Agency (US EPA)-approved west coast marine species for chronic toxicity monitoring and compliance in the National Pollution Discharge Elimination System (NPDES). The protocol allows field-collected sporophylls to be stored for up to 24 h at 9 to 12 C prior to use. However, the effects of sporophyll storage on the bioassay results have not been fully investigated, particularly with kelp collected from beds south of Point Conception, CA, USA. Therefore, 13 matched-pair reference toxicant bioassays using fresh and stored sporophylls collected from a subtidal kelp bed near Laguna Beach, CA, USA, were performed and compared. The results indicate that a lower percentage of spores germinate and the germ tube lengths are reduced when stored sporophylls are used. The intratest precision of the germination endpoint decreased as evidenced by significant increases in the percent minimum significant difference (%MSD) statistic. The intertest precision also decreased in the germination endpoint as demonstrated by significant increases in the coefficient of variation (CV) values at four effect levels. Conversely, a significant reduction in the CVs was observed in the germ tube length data, possibly as a consequence of the decrease in germ tube length associated with storage. Finally, significant decreases in mean effect concentrations in the germination endpoint in tests using stored sporophylls indicated that storage increased the sensitivity of the spores to the toxic effects of CuCl{sub 2}. The toxicological sensitivity and intratest precision of the germ tube length endpoint were not significantly affected by storage of the sporophylls. The effects of sporophyll storage resulted in a high frequency of invalid tests, lower statistical power, less effective quality assurance standards, and apparent bias in the observed toxicity of CuCl{sub 2}.

  17. An Alternative In Vivo Method to Refine the Mouse Bioassay for Botulinum Toxin Detection

    PubMed Central

    Wilder-Kofie, Temeri D; Lúquez, Carolina; Adler, Michael; Dykes, Janet K; Coleman, JoAnn D; Maslanka, Susan E

    2011-01-01

    Botulism is a rare, life-threatening paralytic disease of both humans and animals that is caused by botulinum neurotoxins (BoNT). Botulism is confirmed in the laboratory by the detection of BoNT in clinical specimens, contaminated foods, and cultures. Despite efforts to develop an in vitro method for botulinum toxin detection, the mouse bioassay remains the standard test for laboratory confirmation of this disease. In this study, we evaluated the use of a nonlethal mouse toe-spread reflex model to detect BoNT spiked into buffer, serum, and milk samples. Samples spiked with toxin serotype A and nontoxin control samples were injected into the left and right extensor digitorum longus muscles, respectively. Digital photographs at 0, 8, and 24 h were used to obtain objective measurements through effective paralysis scores, which were determined by comparing the width-to-length ratio between right and left feet. Both objective measurements and clinical observation could accurately identify over 80% of animals injected with 1 LD50 (4.3 pg) BoNT type A within 24 h. Half of animals injected with 0.5 LD50 BoNT type A and none injected with 0.25 LD50 demonstrated localized paralysis. Preincubating the toxin with antitoxin prevented the development of positive effective paralysis scores, demonstrating that (1) the effect was specific for BoNT and (2) identification of toxin serotype could be achieved by using this method. These results suggest that the mouse toe-spread reflex model may be a more humane alternative to the current mouse bioassay for laboratory investigations of botulism. PMID:21819693

  18. Chronic bioassays of chlorinated humic acids in B6C3F1 mice

    SciTech Connect

    van Duuren, B.L.; Melchionne, S.; Seidman, I.; Pereira, M.A.

    1986-11-01

    Humic acids (Fluka), chlorinated to carbon:chlorine (C:Cl) ratios of 1:1 and 1:0.3, were administered to B6C3F1 mice, 50 males and 50 females per group, in the drinking water at a total organic carbon (TOC) level of 0.5 g/L. The mice were 6 to 8 weeks old at the beginning of the bioassays. The doses used were based on short-term (8 weeks) evaluations for toxicity, palatability, and weight gain. The chronic bioassays included the following control groups: unchlorinated humic acids (0.5 g/L), no-treatment (100 males and 100 females), dibromoethane (DBE, 2.0 mM in drinking water; positive control) and 0.44% sodium chloride in drinking water, i.e., at the same concentration as those receiving chlorinated humic acids. The chlorinated humic acids were prepared freshly and chemically assayed once per week. All chemicals were, with the exception of DBE, administered for 24 months; DBE was administered for 18 months. The volumes of solutions consumed were measured once weekly. All treatment groups showed normal weight gain except the DBE group. No markedly significant increases in tumor incidences were evident in any of the organs and tissues examined in the chlorinated humic acid groups compared to unchlorinated humic acids and the no-treatment control groups. DBE caused the expected high incidence of squamous carcinomas of the forestomach. The chlorinated humic acids tested contained direct-acting alkylating agents, based on their reactivity with p-nitrobenzylpyridine (PNBP), and showed mutagenic activity in S. typhimurium.

  19. Chronological supplement to the Carcinogenic Potency Database: standardized results of animal bioassays published through December 1982.

    PubMed Central

    Gold, L S; de Veciana, M; Backman, G M; Magaw, R; Lopipero, P; Smith, M; Blumenthal, M; Levinson, R; Bernstein, L; Ames, B N

    1986-01-01

    This paper is a chronological supplement to our earlier publication, "A Carcinogenic Potency Database of the Standardized Results of Animal Bioassays.¿ We report here results of carcinogenesis bioassays published in Technical Reports of the National Cancer Institute/National Toxicology Program between July 1980 and December 1982, and the general literature between July 1981 and December 1982. This supplement includes results of 280 long-term, chronic experiments of 114 test compounds, and reports the same information about each experiment in the same plot format as the earlier paper: e.g., the species and strain of test animal, the route and duration of compound administration, dose level and other aspects of experimental protocol, histopathology and tumor incidence, TD50 and its statistical significance, dose response, author's opinion about carcinogenicity, and literature reference. While a number of appendices are provided to facilitate use of this supplement, we have not duplicated here the material published earlier. Instead, we refer the reader to the earlier publications (Peto et al. and Gold et al.) for a thorough description of the numerical index of carcinogenic potency (TD50), a guide to the plot of the database, and a discussion of the sources of data, the rationale for the inclusion of particular experiments and particular target sites, and the conventions adopted in summarizing the literature. For 44 of the 114 chemicals reported in this second plot, results of earlier experiments are also given in the first plot; since only 1981-1982 results are reported here, the first plot is required for these repeated compounds. In this paper we also give corrections for errors that appeared in the earlier publication. PMID:3530736

  20. Bioassays and selected chemical analysis of biocide-free antifouling coatings.

    PubMed

    Watermann, B T; Daehne, B; Sievers, S; Dannenberg, R; Overbeke, J C; Klijnstra, J W; Heemken, O

    2005-09-01

    Over the years several types of biocide-free antifouling paints have entered the market. The prohibition of biocidal antifouling paints in special areas of some European countries such as Sweden, Denmark and Germany has favoured the introduction of these paints to the market. Several types of biocide-free antifouling paints were subjected to bioassays and selected chemical analysis of leachate and incorporated substances. Both non-eroding coatings (silicones, fibre coats, epoxies, polyurethane, polyvinyl) and eroding coatings (SPCs, ablative) were tested to exclude the presence of active biocides and dangerous compounds. The paints were subjected to the luminescent bacteria test and the cypris larvae settlement assay, the latter delivering information on toxicity as well as on efficacy. The following chemical analyses of selected compounds of dry-film were performed: The results of the bioassays indicated that none of the coatings analysed contained leachable biocides. Nevertheless, some products contained or leached dangerous compounds. The analyses revealed leaching of nonylphenol (up to 74.7 ng/cm2/d after 48 h) and bisphenol A (up to 2.77 ng/cm2/d after 24 h) from epoxy resins used as substitutes for antifouling paints. The heavy metal, zinc, was measured in dry paint film in quantities up to 576,000 ppm in erodable coatings, not incorporated as a biocide but to control the rate of erosion. Values for TBT in silicone elutriates were mostly below the detection limit of 0.005 mg/kg. Values for DBT ranged between <0.005 and 6.28 mg/kg, deriving from catalysts used as curing agents. Some biocide-free paints contained leachable, toxic and dangerous compounds in the dry film, some of which may act as substitutes for biocides or are incorporated as plasticizers or catalysts. Implications to environmental requirements and legislation are discussed. PMID:15878605