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Sample records for 96-well microplate reader

  1. Enzyme activity assay of glycoprotein enzymes based on a boronate affinity molecularly imprinted 96-well microplate.

    PubMed

    Bi, Xiaodong; Liu, Zhen

    2014-12-16

    Enzyme activity assay is an important method in clinical diagnostics. However, conventional enzyme activity assay suffers from apparent interference from the sample matrix. Herein, we present a new format of enzyme activity assay that can effectively eliminate the effects of the sample matrix. The key is a 96-well microplate modified with molecularly imprinted polymer (MIP) prepared according to a newly proposed method called boronate affinity-based oriented surface imprinting. Alkaline phosphatase (ALP), a glycoprotein enzyme that has been routinely used as an indicator for several diseases in clinical tests, was taken as a representative target enzyme. The prepared MIP exhibited strong affinity toward the template enzyme (with a dissociation constant of 10(-10) M) as well as superb tolerance for interference. Thus, the enzyme molecules in a complicated sample matrix could be specifically captured and cleaned up for enzyme activity assay, which eliminated the interference from the sample matrix. On the other hand, because the boronate affinity MIP could well retain the enzymatic activity of glycoprotein enzymes, the enzyme captured by the MIP was directly used for activity assay. Thus, additional assay time and possible enzyme or activity loss due to an enzyme release step required by other methods were avoided. Assay of ALP in human serum was successfully demonstrated, suggesting a promising prospect of the proposed method in real-world applications.

  2. Cellphone-based hand-held microplate reader for point-of-care ELISA testing (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Berg, Brandon; Cortazar, Bingen; Tseng, Derek; Ozkan, Haydar; Feng, Steve; Wei, Qingshan; Chan, Raymond Y.; Burbano, Jordi; Farooqui, Qamar; Lewinski, Michael; Di Carlo, Dino; Garner, Omai B.; Ozcan, Aydogan

    2016-03-01

    Enzyme-linked immunosorbent assay (ELISA) in a microplate format has been a gold standard first-line clinical test for diagnosis of various diseases including infectious diseases. However, this technology requires a relatively large and expensive multi-well scanning spectrophotometer to read and quantify the signal from each well, hindering its implementation in resource-limited-settings. Here, we demonstrate a cost-effective and handheld smartphone-based colorimetric microplate reader for rapid digitization and quantification of immunoserology-related ELISA tests in a conventional 96-well plate format at the point of care (POC). This device consists of a bundle of 96 optical fibers to collect the transmitted light from each well of the microplate and direct all the transmission signals from the wells onto the camera of the mobile-phone. Captured images are then transmitted to a remote server through a custom-designed app, and both quantitative and qualitative diagnostic results are returned back to the user within ~1 minute per 96-well plate by using a machine learning algorithm. We tested this mobile-phone based micro-plate reader in a clinical microbiology lab using FDA-approved mumps IgG, measles IgG, and herpes simplex virus IgG (HSV-1 and HSV-2) ELISA tests on 1138 remnant patient samples (roughly 50% training and 50% testing), and achieved an overall accuracy of ~99% or higher for each ELISA test. This handheld and cost-effective platform could be immediately useful for large-scale vaccination monitoring in low-infrastructure settings, and also for other high-throughput disease screening applications at POC.

  3. Novel versatile smart phone based Microplate readers for on-site diagnoses.

    PubMed

    Fu, Qiangqiang; Wu, Ze; Li, Xiuqing; Yao, Cuize; Yu, Shiting; Xiao, Wei; Tang, Yong

    2016-07-15

    Microplate readers are important diagnostic instruments, used intensively for various readout test kits (biochemical analysis kits and ELISA kits). However, due to their expensive and non-portability, commercial microplate readers are unavailable for home testing, community and rural hospitals, especially in developing countries. In this study, to provide a field-portable, cost-effective and versatile diagnostic tool, we reported a novel smart phone based microplate reader. The basic principle of this devise relies on a smart phone's optical sensor that measures transmitted light intensities of liquid samples. To prove the validity of these devises, developed smart phone based microplate readers were applied to readout results of various analytical targets. These targets included analanine aminotransferase (ALT; limit of detection (LOD) was 17.54 U/L), alkaline phosphatase (AKP; LOD was 15.56 U/L), creatinine (LOD was 1.35μM), bovine serum albumin (BSA; LOD was 0.0041mg/mL), prostate specific antigen (PSA; LOD was 0.76pg/mL), and ractopamine (Rac; LOD was 0.31ng/mL). The developed smart phone based microplate readers are versatile, portable, and inexpensive; they are unique because of their ability to perform under circumstances where resources and expertize are limited.

  4. High-throughput measurements of the optical redox ratio using a commercial microplate reader

    NASA Astrophysics Data System (ADS)

    Cannon, Taylor M.; Shah, Amy T.; Walsh, Alex J.; Skala, Melissa C.

    2015-01-01

    There is a need for accurate, high-throughput, functional measures to gauge the efficacy of potential drugs in living cells. As an early marker of drug response in cells, cellular metabolism provides an attractive platform for high-throughput drug testing. Optical techniques can noninvasively monitor NADH and FAD, two autofluorescent metabolic coenzymes. The autofluorescent redox ratio, defined as the autofluorescence intensity of NADH divided by that of FAD, quantifies relative rates of cellular glycolysis and oxidative phosphorylation. However, current microscopy methods for redox ratio quantification are time-intensive and low-throughput, limiting their practicality in drug screening. Alternatively, high-throughput commercial microplate readers quickly measure fluorescence intensities for hundreds of wells. This study found that a commercial microplate reader can differentiate the receptor status of breast cancer cell lines (p<0.05) based on redox ratio measurements without extrinsic contrast agents. Furthermore, microplate reader redox ratio measurements resolve response (p<0.05) and lack of response (p>0.05) in cell lines that are responsive and nonresponsive, respectively, to the breast cancer drug trastuzumab. These studies indicate that the microplate readers can be used to measure the redox ratio in a high-throughput manner and are sensitive enough to detect differences in cellular metabolism that are consistent with microscopy results.

  5. High-throughput measurements of the optical redox ratio using a commercial microplate reader

    PubMed Central

    Cannon, Taylor M.; Shah, Amy T.; Walsh, Alex J.; Skala, Melissa C.

    2015-01-01

    Abstract. There is a need for accurate, high-throughput, functional measures to gauge the efficacy of potential drugs in living cells. As an early marker of drug response in cells, cellular metabolism provides an attractive platform for high-throughput drug testing. Optical techniques can noninvasively monitor NADH and FAD, two autofluorescent metabolic coenzymes. The autofluorescent redox ratio, defined as the autofluorescence intensity of NADH divided by that of FAD, quantifies relative rates of cellular glycolysis and oxidative phosphorylation. However, current microscopy methods for redox ratio quantification are time-intensive and low-throughput, limiting their practicality in drug screening. Alternatively, high-throughput commercial microplate readers quickly measure fluorescence intensities for hundreds of wells. This study found that a commercial microplate reader can differentiate the receptor status of breast cancer cell lines (p<0.05) based on redox ratio measurements without extrinsic contrast agents. Furthermore, microplate reader redox ratio measurements resolve response (p<0.05) and lack of response (p>0.05) in cell lines that are responsive and nonresponsive, respectively, to the breast cancer drug trastuzumab. These studies indicate that the microplate readers can be used to measure the redox ratio in a high-throughput manner and are sensitive enough to detect differences in cellular metabolism that are consistent with microscopy results. PMID:25634108

  6. Development of LEDs-based microplate reader for bioanalytical assay measurements

    NASA Astrophysics Data System (ADS)

    Alaruri, Sami D.; Katzlinger, Michael; Schinwald, Bernhard; Kronberger, Georg; Atzler, Joseph

    2013-10-01

    The optical design for an LEDs-based microplate reader that can perform fluorescence intensity (top and bottom), absorbance, luminescence and time-resolved fluorescence measurements is described. The microplate reader is the first microplate reader in the marketplace that incorporates LEDs as excitation light sources. Absorbance measurements over the 0-3.5 optical density range for caffeine solution are presented. Additionally, fluorescence intensity readings collected at 535 and 625 nm from a green and a red RediPlateTM are reported. Furthermore, fluorescence decay lifetime measurements obtained for Eu (europium) and Sm (samarium) standard solutions using 370 nm excitation are presented. The microplate reader detection limits for the fluorescence intensity top, fluorescence intensity bottom, fluorescence polarization and time-resolved fluorescence modes are 1.5 fmol 100 µL-1 fluorescein (384-well plate), 25 fmol 100 µL-1 fluorescein (384-well plate), 5 mP at 10 nM fluorescein (black 384-well plate) and 30 amol 100 µL-1 europium solution (white 384-well plate), respectively.

  7. Cellphone-Based Hand-Held Microplate Reader for Point-of-Care Testing of Enzyme-Linked Immunosorbent Assays.

    PubMed

    Berg, Brandon; Cortazar, Bingen; Tseng, Derek; Ozkan, Haydar; Feng, Steve; Wei, Qingshan; Chan, Raymond Yan-Lok; Burbano, Jordi; Farooqui, Qamar; Lewinski, Michael; Di Carlo, Dino; Garner, Omai B; Ozcan, Aydogan

    2015-08-25

    Standard microplate based enzyme-linked immunosorbent assays (ELISA) are widely utilized for various nanomedicine, molecular sensing, and disease screening applications, and this multiwell plate batched analysis dramatically reduces diagnosis costs per patient compared to nonbatched or nonstandard tests. However, their use in resource-limited and field-settings is inhibited by the necessity for relatively large and expensive readout instruments. To mitigate this problem, we created a hand-held and cost-effective cellphone-based colorimetric microplate reader, which uses a 3D-printed opto-mechanical attachment to hold and illuminate a 96-well plate using a light-emitting-diode (LED) array. This LED light is transmitted through each well, and is then collected via 96 individual optical fibers. Captured images of this fiber-bundle are transmitted to our servers through a custom-designed app for processing using a machine learning algorithm, yielding diagnostic results, which are delivered to the user within ∼1 min per 96-well plate, and are visualized using the same app. We successfully tested this mobile platform in a clinical microbiology laboratory using FDA-approved mumps IgG, measles IgG, and herpes simplex virus IgG (HSV-1 and HSV-2) ELISA tests using a total of 567 and 571 patient samples for training and blind testing, respectively, and achieved an accuracy of 99.6%, 98.6%, 99.4%, and 99.4% for mumps, measles, HSV-1, and HSV-2 tests, respectively. This cost-effective and hand-held platform could assist health-care professionals to perform high-throughput disease screening or tracking of vaccination campaigns at the point-of-care, even in resource-poor and field-settings. Also, its intrinsic wireless connectivity can serve epidemiological studies, generating spatiotemporal maps of disease prevalence and immunity.

  8. Microplate-reader method for the rapid analysis of copper in natural waters with chemiluminescence detection

    PubMed Central

    Durand, Axel; Chase, Zanna; Remenyi, Tomas; Quéroué, Fabien

    2013-01-01

    We have developed a method for the determination of copper in natural waters at nanomolar levels. The use of a microplate-reader minimizes sample processing time (~25 s per sample), reagent consumption (~120 μL per sample), and sample volume (~700 μL). Copper is detected by chemiluminescence. This technique is based on the formation of a complex between copper and 1,10-phenanthroline and the subsequent emission of light during the oxidation of the complex by hydrogen peroxide. Samples are acidified to pH 1.7 and then introduced directly into a 24-well plate. Reagents are added during data acquisition via two reagent injectors. When trace metal clean protocols are employed, the reproducibility is generally less than 7% on blanks and the detection limit is 0.7 nM for seawater and 0.4 nM for freshwater. More than 100 samples per hour can be analyzed with this technique, which is simple, robust, and amenable to at-sea analysis. Seawater samples from Storm Bay in Tasmania illustrate the utility of the method for environmental science. Indeed other trace metals for which optical detection methods exist (e.g., chemiluminescence, fluorescence, and absorbance) could be adapted to the microplate-reader. PMID:23335917

  9. Validation and characterization of optical redox ratio measurements with a microplate reader in breast cancer cells

    NASA Astrophysics Data System (ADS)

    Cannon, Taylor M.; Shah, Amy T.; Skala, Melissa C.

    2015-02-01

    There is a need for accurate, high-throughput measures to gauge the efficacy of potential drugs in living cells. Metabolism is an early marker of drug response in cells, and NADH and FAD are autofluorescent cellular metabolic coenzymes that can be non-invasively monitored using optical techniques. Relative rates of glycolysis and oxidative phosphorylation in a cell are quantified by the redox ratio, defined as the autofluorescence intensity of NADH divided by that of FAD. Microplate readers are high-throughput instruments that can rapidly measure NADH and FAD autofluorescence intensities for hundreds of wells, and are capable of identifying receptor status and resolving drug response in breast cancer cell lines. This study tests the accuracy and repeatability of plate reader experiments measuring the redox ratio in breast cancer cell lines. NADH and FAD fluorescence levels remained constant over the course of multiple measurements (p<0.1), ruling out the incidence of photobleaching. The contribution of media to background fluorescence signal was also investigated. Media fluorescence levels for both coenzymes were significantly lower (p<0.0001) than those from wells containing cells, and replacing the media with saline resulted in the same redox ratio trends among cell lines as initial measurements with media. Following treatment with carbonyl cyanide p-fluorodeoxyphenylhydrazone (FCCP), an oxidative phosphorylation inhibitor, the redox ratio decreased (p<0.05), validating NADH and FAD as the primary fluorescence sources. These findings verify that autofluorescence measurements taken by microplate readers accurately and reliably characterize NADH and FAD fluorescence, validating their promise in the areas of metabolic monitoring and drug development.

  10. Electrothermal micromixing in 96 well plate

    NASA Astrophysics Data System (ADS)

    Kauffmann, Paul; Loire, Sophie; Mezic, Igor

    2011-11-01

    Diagnostic and pharmacology processes could be greatly accelerated by appropriate mixing. Here electrothermal flows are explored to provide mixing of conductive physiological solutions (=1.6 S/m) in a 96 well plate. Three interdigitated electrodes provide an electric field (< 15Vpp, 1MHz) beneath each well. Polarization and conduction phenomenon of the fluid in a well will be first modeled numerically and compared to an electrical circuit model. Due to high conductivity and permittivity of the fluid, the impedance of the array of filled wells collapse dramatically (96 wells: R = 1Ohm, C=250nF). The power supply challenges accordingly raised by arrays of electrothermal micromixers will be then analyzed. The efficiency of different methods of mixing in those wells will be also compared: the addition of low frequency signal leading to AC electro-osmotic perturbations, a blinking vortices method. The experimental results will be compared to simulations.

  11. High-throughput and automated diagnosis of antimicrobial resistance using a cost-effective cellphone-based micro-plate reader

    NASA Astrophysics Data System (ADS)

    Feng, Steve; Tseng, Derek; di Carlo, Dino; Garner, Omai B.; Ozcan, Aydogan

    2016-12-01

    Routine antimicrobial susceptibility testing (AST) can prevent deaths due to bacteria and reduce the spread of multi-drug-resistance, but cannot be regularly performed in resource-limited-settings due to technological challenges, high-costs, and lack of trained professionals. We demonstrate an automated and cost-effective cellphone-based 96-well microtiter-plate (MTP) reader, capable of performing AST without the need for trained diagnosticians. Our system includes a 3D-printed smartphone attachment that holds and illuminates the MTP using a light-emitting-diode array. An inexpensive optical fiber-array enables the capture of the transmitted light of each well through the smartphone camera. A custom-designed application sends the captured image to a server to automatically determine well-turbidity, with results returned to the smartphone in ~1 minute. We tested this mobile-reader using MTPs prepared with 17 antibiotics targeting Gram-negative bacteria on clinical isolates of Klebsiella pneumoniae, containing highly-resistant antimicrobial profiles. Using 78 patient isolate test-plates, we demonstrated that our mobile-reader meets the FDA-defined AST criteria, with a well-turbidity detection accuracy of 98.21%, minimum-inhibitory-concentration accuracy of 95.12%, and a drug-susceptibility interpretation accuracy of 99.23%, with no very major errors. This mobile-reader could eliminate the need for trained diagnosticians to perform AST, reduce the cost-barrier for routine testing, and assist in spatio-temporal tracking of bacterial resistance.

  12. High-throughput and automated diagnosis of antimicrobial resistance using a cost-effective cellphone-based micro-plate reader

    PubMed Central

    Feng, Steve; Tseng, Derek; Di Carlo, Dino; Garner, Omai B.; Ozcan, Aydogan

    2016-01-01

    Routine antimicrobial susceptibility testing (AST) can prevent deaths due to bacteria and reduce the spread of multi-drug-resistance, but cannot be regularly performed in resource-limited-settings due to technological challenges, high-costs, and lack of trained professionals. We demonstrate an automated and cost-effective cellphone-based 96-well microtiter-plate (MTP) reader, capable of performing AST without the need for trained diagnosticians. Our system includes a 3D-printed smartphone attachment that holds and illuminates the MTP using a light-emitting-diode array. An inexpensive optical fiber-array enables the capture of the transmitted light of each well through the smartphone camera. A custom-designed application sends the captured image to a server to automatically determine well-turbidity, with results returned to the smartphone in ~1 minute. We tested this mobile-reader using MTPs prepared with 17 antibiotics targeting Gram-negative bacteria on clinical isolates of Klebsiella pneumoniae, containing highly-resistant antimicrobial profiles. Using 78 patient isolate test-plates, we demonstrated that our mobile-reader meets the FDA-defined AST criteria, with a well-turbidity detection accuracy of 98.21%, minimum-inhibitory-concentration accuracy of 95.12%, and a drug-susceptibility interpretation accuracy of 99.23%, with no very major errors. This mobile-reader could eliminate the need for trained diagnosticians to perform AST, reduce the cost-barrier for routine testing, and assist in spatio-temporal tracking of bacterial resistance. PMID:27976700

  13. A simple high-throughput method for determination of antiepileptic analogues of γ-aminobutyric acid in pharmaceutical dosage forms using microplate fluorescence reader.

    PubMed

    Martinc, Boštjan; Vovk, Tomaž

    2013-01-01

    Pregabalin (PGB), gabapentin (GBP), and vigabatrin (VGB) are structural analogues of γ-aminobutyric acid used for the treatment of different forms of epilepsy. Their analytical determination is challenging since these molecules have no significant UV or visible absorption. Several derivatization methods have been developed and used for their determination in bulk or pharmaceutical dosage forms. We aimed to develop a high- throughput method using a microplate reader with fluorescence detection and simple derivatization with fluorescamine. Obtained method involves derivatization step of only 5 min at room temperature and simultaneous measurements of 96 samples (λex 395, λem 476 nm) thus rendering excellent high-throughput analysis. The method was found to be linear with r²>0.998 across investigated analytical ranges of 0.75 to 30.0 µg/mL for PGB, 2.00 to 80.0 µg/mL for GBP, and 1.50 to 60.0 µg/mL for VGB. Intraday and interday precision values did not exceed 4.93%. The accuracy was ranging between 96.6 to 103.5%. The method was also found to be specific since used excipients did not interfere with the method. The robustness study showed that derivatization procedure is more robust than spectrofluorimetric conditions. The developed high-throughput method was successfully applied for determination of drug content and dissolution profiles in pharmaceutical dosage forms of studied antiepileptic drugs.

  14. Thermodynamic equilibrium solubility measurements in simulated fluids by 96-well plate method in early drug discovery.

    PubMed

    Bharate, Sonali S; Vishwakarma, Ram A

    2015-04-01

    An early prediction of solubility in physiological media (PBS, SGF and SIF) is useful to predict qualitatively bioavailability and absorption of lead candidates. Despite of the availability of multiple solubility estimation methods, none of the reported method involves simplified fixed protocol for diverse set of compounds. Therefore, a simple and medium-throughput solubility estimation protocol is highly desirable during lead optimization stage. The present work introduces a rapid method for assessment of thermodynamic equilibrium solubility of compounds in aqueous media using 96-well microplate. The developed protocol is straightforward to set up and takes advantage of the sensitivity of UV spectroscopy. The compound, in stock solution in methanol, is introduced in microgram quantities into microplate wells followed by drying at an ambient temperature. Microplates were shaken upon addition of test media and the supernatant was analyzed by UV method. A plot of absorbance versus concentration of a sample provides saturation point, which is thermodynamic equilibrium solubility of a sample. The established protocol was validated using a large panel of commercially available drugs and with conventional miniaturized shake flask method (r(2)>0.84). Additionally, the statistically significant QSPR models were established using experimental solubility values of 52 compounds.

  15. An optical microbial biosensor for detection of methyl parathion using Sphingomonas sp. immobilized on microplate as a reusable biocomponent.

    PubMed

    Kumar, Jitendra; D'Souza, S F

    2010-12-15

    Organophosphorus pesticides such as methyl parathion have been widely used in the field of agriculture for insect pest control. These pesticides and their degradation products cause environmental pollution and ecological problem. With a view to monitor these pesticides biosensors are being developed. A bacterium Sphingomonas sp. from field soil has been isolated and identified in our laboratory that hydrolyzes the methyl parathion upto a chromophoric product, p-nitrophenol (PNP). PNP can be detected by electrochemical and colorimetric methods, which can be exploited to develop a biosensor for detection of the organophosphate pesticide. Whole cells of Sphingomonas bacteria were immobilized directly onto the surface of the wells of polystyrene microplates (96 wells) using glutaraldehyde as the cross-linker. SEM study confirmed the immobilization of Sphingomonas sp. Immobilized bacterial microplate was associated directly with the optical transducer, microplate reader. The microplate-based biosensor is having advantages as it has 96 reaction vessels and therefore it provides a convenient system for detecting multiple numbers of samples in a single platform. Detection range of the biosensor from the linear range was determined to be 4-80 μM methyl parathion. Cells-immobilized microplates were having reusability upto 75 reactions. Present study reports an innovative concept where the microplate can be used as immobilizing support for development of reusable microbial biocomponent.

  16. An automated 96-well-plate loader for FACScan.

    PubMed

    Illges, H

    1999-01-01

    Staining multiple samples for data acquisition with a flow cytometer is done in 96-well plates to save time and make large data assessment in one working day possible. However, the inability of the FACScan to take up the samples from 96-well plates is a major drawback. In order to avoid the individual transfer of samples to tubes, we have developed a system, which allows using the FACScan with 96-well plates. The machine consists of a programmable control module and a loader which moves the 96-well plate in 3 axis well by well along the sample collector. The machine is equipped with a wash buffer tank to avoid cross contamination of samples and a shaking option to avoid sedimentation of cells during acquisition. The machine can be further developed into a full automatic loader if connected to the FACS Station. In 24 hours about 7,000 samples with 10,000 cells each can be acquired.

  17. 96-well electroporation method for transfection of mammalian central neurons.

    PubMed

    Buchser, William J; Pardinas, Jose R; Shi, Yan; Bixby, John L; Lemmon, Vance P

    2006-11-01

    Manipulating gene expression in primary neurons has been a goal for many scientists for over 20 years. Vertebrate central nervous system neurons are classically difficult to transfect. Most lipid reagents are inefficient and toxic to the cells, and time-consuming methods such as viral infections are often required to obtain better efficiencies. We have developed an efficient method for the transfection of cerebellar granule neurons and hippocampal neurons with standard plasmid vectors. Using 96-well electroporation plates, square-wave pulses can introduce 96 different plasmids into neurons in a single step. The procedure results in greater than 20% transfection efficiencies and requires only simple solutions of nominal cost. In addition to enabling the rapid optimization of experimental protocols with multiple parameters, this procedure enables the use of high content screening methods to characterize neuronal phenotypes.

  18. 96-Well electroporation method for transfection of mammalian central neurons

    PubMed Central

    Buchser, William J.; Pardinas, Jose R.; Shi, Yan; Bixby, John L.; Lemmon, Vance P.

    2008-01-01

    Manipulating gene expression in primary neurons has been a goal for many scientists for over 20 years. Vertebrate central nervous system neurons are classically difficult to transfect. Most lipid reagents are inefficient and toxic to the cells, and time-consuming methods such as viral infections are often required to obtain better efficiencies. We have developed an efficient method for the transfection of cerebellar granule neurons and hippocampal neurons with standard plasmid vectors. Using 96-well electroporation plates, square-wave pulses can introduce 96 different plasmids into neurons in a single step. The procedure results in greater than 20% transfection efficiencies and requires only simple solutions of nominal cost. In addition to enabling the rapid optimization of experimental protocols with multiple parameters, this procedure enables the use of high content screening methods to characterize neuronal phenotypes. PMID:17140120

  19. 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…

  20. Nitrotyrosine Density of Rabbit Urinary Bladder Muscle and Mucosa Measured via Western Blotting and 96-Well Plate Analysis.

    PubMed

    Fitzpatrick, Brittany; Schuler, Catherine; Leggett, Robert E; Levin, Robert M

    2012-01-01

    Purpose. Nitrotyrosine was quantitated in rabbit bladder muscle and mucosa using two analytical systems: Western blotting analyses and a 96-well plate quantitative analysis kit. Materials and Methods. Rabbit bladder muscle and mucosa were obtained from control rabbits. For the Western analysis, the samples were loaded into a SDS page gel and then transferred to a PVDF membrane. The optical density was measured using a Kodak Scanner. Using the 96-well plate, the samples and standards were loaded, incubated with primary and secondary antibody, washed and vacuumed with 10x wash buffer three times between each incubation period. Stop buffer was added to the plate and the results were quantified via the plate reader. Results. For both muscle and mucosa tissue, the optical density readings were linear with tissue concentration; the concentration of nitrotyrosine in the mucosa was significantly higher than in the muscle. However, whereas the Western blot analysis is based on relative optical densities, the 96-well plate kit provides a truly quantitative analysis. Discussion. Mucosa tissue displayed a higher density of nitrotyrosine than did detrusor muscle tissue. This may well be due to the significantly higher metabolic activity of the mucosa compared to the muscle.

  1. Enhancement of Fluorescence-Based Sandwich Immunoassay Using Multilayered Microplates Modified with Plasma-Polymerized Films.

    PubMed

    Yano, Kazuyoshi; Iwasaki, Akira

    2016-12-25

    A functional modification of the surface of a 96-well microplate coupled with a thin layer deposition technique is demonstrated for enhanced fluorescence-based sandwich immunoassays. The plasma polymerization technique enabling the deposition of organic thin films was employed for the modification of the well surface of a microplate. A silver layer and a plasma-polymerized film were consecutively deposited on the microplate as a metal mirror and the optical interference layer, respectively. When Cy3-labeled antibody was applied to the wells of the resulting multilayered microplate without any immobilization step, greatly enhanced fluorescence was observed compared with that obtained with the unmodified one. The same effect could be also exhibited for an immunoassay targeting antigen directly adsorbed on the multilayered microplate. Furthermore, a sandwich immunoassay for the detection of interleukin 2 (IL-2) was performed with the multilayered microplates, resulting in specific and 88-fold-enhanced fluorescence detection.

  2. Enhancement of Fluorescence-Based Sandwich Immunoassay Using Multilayered Microplates Modified with Plasma-Polymerized Films

    PubMed Central

    Yano, Kazuyoshi; Iwasaki, Akira

    2016-01-01

    A functional modification of the surface of a 96-well microplate coupled with a thin layer deposition technique is demonstrated for enhanced fluorescence-based sandwich immunoassays. The plasma polymerization technique enabling the deposition of organic thin films was employed for the modification of the well surface of a microplate. A silver layer and a plasma-polymerized film were consecutively deposited on the microplate as a metal mirror and the optical interference layer, respectively. When Cy3-labeled antibody was applied to the wells of the resulting multilayered microplate without any immobilization step, greatly enhanced fluorescence was observed compared with that obtained with the unmodified one. The same effect could be also exhibited for an immunoassay targeting antigen directly adsorbed on the multilayered microplate. Furthermore, a sandwich immunoassay for the detection of interleukin 2 (IL-2) was performed with the multilayered microplates, resulting in specific and 88-fold–enhanced fluorescence detection. PMID:28029144

  3. Development of a fluorescence-based microplate method for the determination of volatile fatty acids in anaerobically digested and sewage sludges.

    PubMed

    Robert-Peillard, F; Palacio-Barco, E; Coulomb, B; Boudenne, J L

    2012-01-15

    This paper presents a simple, accurate and multi-sample method for the determination of volatile fatty acids (VFAs) thanks to a 96-well microplate technique. A procedure using an activating reagent of the carboxylic function (water-soluble carbodiimide EDC) and a fluorescent amino labeling reagent (N-(1-naphthyl)ethylenediamine, EDAN) allows the formation of an isoindole derivative that needs to be separated from initial fluorescent amine for efficient VFAs determination. Isolation of these fluorescent VFA-derivatives was carried out by use of the fluorescent quenching of EDAN with o-phthaldialdehyde (OPA). Quenching was most efficient at pH around 7 and by heating at 40°C within the microplate reader. This optimized procedure has been applied to various carboxylic acids and other organic compounds, demonstrating that VFA exhibit the highest fluorescence responses with homogeneous results for the main ones (acetic, propionic and butyric acid, all mass concentration expressed as acetic acid equivalents). This protocol was calibrated against acetic acid and determination of VFA was thus possible in the range 3.9-2,000 mg L(-1) (acetic acid equivalents). Subsequent application to real samples (sewage sludges or anaerobically digested samples) and comparison to gas chromatography analyses gave accurate results, proving the great potential of our high-throughput microplate-based technique for the analysis of VFA.

  4. Microplates with adaptive surfaces.

    PubMed

    Akbulut, Meshude; Lakshmi, Dhana; Whitcombe, Michael J; Piletska, Elena V; Chianella, Iva; Güven, Olgun; Piletsky, Sergey A

    2011-11-14

    Here we present a new and versatile method for the modification of the well surfaces of polystyrene microtiter plates (microplates) with poly(N-phenylethylene diamine methacrylamide), (poly-NPEDMA). The chemical grafting of poly-NPEDMA to the surface of microplates resulted in the formation of thin layers of a polyaniline derivative bearing pendant methacrylamide double bonds. These were used as the attachment point for various functional polymers through photochemical grafting of various, for example, acrylate and methacrylate, polymers with different functionalities. In a model experiment, we have modified poly-NPEDMA-coated microplates with a small library of polymers containing different functional groups using a two-step approach. In the first step, double bonds were activated by UV irradiation in the presence of N,N-diethyldithiocarbamic acid benzyl ester (iniferter). This enabled grafting of the polymer library in the second step by UV irradiation of solutions of the corresponding monomers in the microplate wells. The uniformity of coatings was confirmed spectrophotometrically, by microscopic imaging and by contact angle measurements (CA). The feasibility of the current technology has been shown by the generation of a small library of polymers grafted to the microplate well surfaces and screening of their affinity to small molecules, such as atrazine, a trio of organic dyes, and a model protein, bovine serum albumin (BSA). The stability of the polymers, reproducibility of measurement, ease of preparation, and cost-effectiveness make this approach suitable for applications in high-throughput screening in the area of materials research.

  5. Calcofluor fluorescence assay for wort beta-glucan in a microplate format

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The widely-used fluorescent (Calcofluor) flow injection analysis method for determining the concentrations of beta-glucans in Congress worts from barley malts is adapted to microplate format. Adaptation of the Calcofluor assay to use widely available fluorescent microplate readers makes the assay m...

  6. Tectonic microplates: laying it down on wax

    NASA Astrophysics Data System (ADS)

    Katz, R. R.; Bodenschatz, E.

    2008-12-01

    We present a wax analogue model of sea-floor spreading that produces rotating, growing microplates. Wax microplates are kinematically similar to sea-floor tectonic microplates in terms of spreading rate and growth rate. Furthermore, their spiral pseudofault geometry is quantitatively consistent with Schouten's oceanic microplate model. These results suggest that Schouten's edge-driven microplate model captures the kinematics of tectonic microplate evolution on Earth. We propose a theory for the formation of microplates.

  7. HIGHLY SENSITIVE ASSAY FOR ANTICHOLINESTERASE COMPOUNDS USING 96 WELL PLATE FORMAT

    EPA Science Inventory

    The rapid and sensitive detection of organophosphate insecticides using a 96 well plate format is reported. Several features of this assay make it attractive for development as a laboratory-based or field screening assay. Acetylcholinesterase (AChE) was stabilized in a gelati...

  8. An improved 96-well turbidity assay for T4 lysozyme activity

    PubMed Central

    Toro, Tasha B.; Nguyen, Thao P.; Watt, Terry J.

    2015-01-01

    T4 lysozyme (T4L) is an important model system for investigating the relationship between protein structure and function. Despite being extensively studied, a reliable, quantitative activity assay for T4L has not been developed. Here, we present an improved T4L turbidity assay as well as an affinity-based T4L expression and purification protocol. This assay is designed for 96-well format and utilizes conditions amenable for both T4L and other lysozymes. This protocol enables easy, efficient, and quantitative characterization of T4L variants and allows comparison between different lysozymes. Our method: • Is applicable for all lysozymes, with enhanced sensitivity for T4 lysozyme compared to other 96-well plate turbidity assays; • Utilizes standardized conditions for comparing T4 lysozyme variants and other lysozymes; and • Incorporates a simplified expression and purification protocol for T4 lysozyme. PMID:26150996

  9. 96-Well plate assays for measuring collagenase activity using (3)H-acetylated collagen.

    PubMed

    Koshy, P J; Rowan, A D; Life, P F; Cawston, T E

    1999-11-15

    We describe two alternative assays for measuring collagenolytic activity using (3)H-acetylated collagen. Both assays have been developed for the 96-well plate format and measure the amount of radiolabeled collagen fragments released into the supernatant from an insoluble (3)H-acetylated collagen fibril preparation. The first method separates digested solubilized fragments from the intact fibril by sedimentation of the undigested collagen by centrifugation. The second method achieves this separation by filtration of the supernatant through the membrane of a 96-well filtration plate which retains the undigested collagen fibril. Both methods give linear dose- and time-dependent responses of collagenase activity > or = 70% of total collagen lysis. In addition, both assays can be simply modified to measure tissue inhibitors of metalloproteinases (TIMPs) inhibitory activity, which is also linear between 20 and 75% of total collagen lysis with the amount of TIMP added.

  10. Rapid evaluation of oxygen and water permeation through microplate sealing tapes.

    PubMed

    Zimmermann, Hartmut F; John, Gernot T; Trauthwein, Harald; Dingerdissen, Uwe; Huthmacher, Klaus

    2003-01-01

    Eight commercially available microplate sealing tapes and 10 other suitable materials (transparent wound dressings) are compared qualitatively in terms of their ability to minimize water evaporation from a multiwell plate while maintaining the oxygen supply as high as possible, which is necessary for applications like aerobic growth. The transparency and sterility of the products are considered as well. All evaluated commercially available sealing tapes fall into one of the following two classes: (1) O(2) transfer is comparable to that of an unsealed plate, but water vapor retention is relatively low, or (2) O(2) transfer via the sealing is slower, but the water retention capability is comparably high. All but one of the evaluated wound dressings fall under the second class. That dressing, however, constitutes a compromise by showing both moderate O(2) permeability and medium water retention. But the estimated mass transport in a microtiter plate sealed with this dressing is about 5 times slower than that of an unsealed 96 well plate. The aim of this publication is to enable the reader to choose a microtiter plate sealing from the materials evaluated within this work and to use the rapid methods described herein to easily perform tests of additional sealing materials.

  11. A 96-well plate assay for CYP4503A induction using cryopreserved human hepatocytes.

    PubMed

    Kamiguchi, Naomi; Aoyama, Eiji; Okuda, Teruaki; Moriwaki, Toshiya

    2010-11-01

    A reliable and practical CYP3A induction assay with cryopreserved human hepatocytes in a 96-well format was developed. Various 96-well plates with different basement membrane were evaluated using prototypical inducers, rifampicin, phenytoin, and carbamazepine. Thin-layer (TL) Matrigel was found to yield the highest basal and induced levels of CYP3A activity as determined by testosterone 6β-hydroxylation. Concentration-dependent CYP3A induction of rifampicin was reproducible with the EC(50) values of 0.36 ± 0.28 μM from four batches of human hepatocytes using the 96-well plate with TL Matrigel. The rank order of induction potency for nine inducers or noninducers at a concentration of 10 μM were well comparable among the multiple donors, by expressing the results as percentage of change compared with the positive control, 10 μM rifampicin. Cotreatment of avasimibe or efavirenz with 10 μM rifampicin was found to reduce CYP3A activities induced by rifampicin at a lower rate than treatment with rifampicin alone, whereas treatment with phenobarbital and carbamazepine had no effect. From a comparison of induced CYP3A activities and gene expression levels, there were compounds that would cause induction of CYP3A4 mRNA but not activity, presumably due to their inhibitory effect on CYP3A activity. The cotreatment assay of test compound with rifampicin allows us to exclude the false-negative results caused by the cytotoxicity and/or the mechanism-based inactivation, when the drug candidate's ability for CYP3A induction is evaluating the enzyme activity. This 96-well plate assay, which is robust, reproducible, and convenient, has demonstrated the paramount applicability to the early drug discovery stage.

  12. FIPRONIL EFFECTS ON ESTUARINE COPEPOD (AMPHIASCUS TENUIREMIS) DEVELOPMENT, FERTILITY, AND REPRODUCTION: A RAPID LIFE-CYCLE ASSAY IN 96-WELL MICROPLATE FORMAT. (R827397)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  13. Magnetochromatic thin-film microplates.

    PubMed

    He, Le; Janner, Michael; Lu, Qipeng; Wang, Mingsheng; Ma, Hua; Yin, Yadong

    2015-01-07

    A new type of magnetochromatic material is developed based on thin-film interference of microplates self-assembled from super-paramagnetic nanocrystals. Dynamic optical tuning can be achieved through orientational manipulation of free-standing super-paramagnetic thin-film microplates using external magnetic fields.

  14. Stretch Injury of Human Induced Pluripotent Stem Cell Derived Neurons in a 96 Well Format

    PubMed Central

    Sherman, Sydney A.; Phillips, Jack K.; Costa, J. Tighe; Cho, Frances S.; Oungoulian, Sevan R.; Finan, John D.

    2016-01-01

    Traumatic brain injury (TBI) is a major cause of mortality and morbidity with limited therapeutic options. Traumatic axonal injury (TAI) is an important component of TBI pathology. It is difficult to reproduce TAI in animal models of closed head injury, but in vitro stretch injury models reproduce clinical TAI pathology. Existing in vitro models employ primary rodent neurons or human cancer cell line cells in low throughput formats. This in vitro neuronal stretch injury model employs human induced pluripotent stem cell-derived neurons (hiPSCNs) in a 96 well format. Silicone membranes were attached to 96 well plate tops to create stretchable, culture substrates. A custom-built device was designed and validated to apply repeatable, biofidelic strains and strain rates to these plates. A high content approach was used to measure injury in a hypothesis-free manner. These measurements are shown to provide a sensitive, dose-dependent, multi-modal description of the response to mechanical insult. hiPSCNs transition from healthy to injured phenotype at approximately 35% Lagrangian strain. Continued development of this model may create novel opportunities for drug discovery and exploration of the role of human genotype in TAI pathology. PMID:27671211

  15. Edge-driven microplate kinematics

    USGS Publications Warehouse

    Schouten, Hans; Klitgord, Kim D.; Gallo, David G.

    1993-01-01

    It is known from plate tectonic reconstructions that oceanic microplates undergo rapid rotation about a vertical axis and that the instantaneous rotation axes describing the microplate's motion relative to the bounding major plates are frequently located close to its margins with those plates, close to the tips of propagating rifts. We propose a class of edge-driven block models to illustrate how slip across the microplate margins, block rotation, and propagation of rifting may be related to the relative motion of the plates on either side. An important feature of these edge-driven models is that the instantaneous rotation axes are always located on the margins between block and two bounding plates. According to those models the pseudofaults or traces of disrupted seafloor resulting from the propagation of rifting between microplate and major plates may be used independently to approximately trace the continuous kinematic evolution of the microplate back in time. Pseudofault geometries and matching rotations of the Easter microplate show that for most of its 5 m.y. history, block rotation could be driven by the drag of the Nazca and Pacific plates on the microplate's edges rather than by a shear flow of mantle underneath.

  16. High throughput preparation of fly genomic DNA in 96-well format using a paint-shaker.

    PubMed

    Lang, Michael; Nagy, Olga; Lang, Claus; Orgogozo, Virginie

    2015-01-01

    Sample homogenization is an essential step for genomic DNA extraction, with multiple downstream applications in Molecular Biology. Genotyping hundreds or thousands of samples requires an automation of this homogenization step, and high throughput homogenizer equipment currently costs 7000 euros or more. We present an apparatus for homogenization of individual Drosophila adult flies in 96-well micro-titer dishes, which was built from a small portable paint-shaker (F5 portable paint-shaker, Ushake). Single flies are disrupted in each well that contains extraction buffer and a 4-mm metal ball. Our apparatus can hold up to five 96-well micro-titer plates. Construction of the homogenizer apparatus takes about 3-4 days, and all equipment can be obtained from a home improvement store. The total material cost is approximately 700 euros including the paint-shaker. We tested the performance of our apparatus using the ZR-96 Quick-gDNA™ kit (Zymo Research) homogenization buffer and achieved nearly complete tissue homogenization after 15 minutes of shaking. PCR tests did not detect any cross contamination between samples of neighboring wells. We obtained on average 138 ng of genomic DNA per fly, and DNA quality was adequate for standard PCR applications. In principle, our tissue homogenizer can be used for isolation of DNA suitable for library production and high throughput genotyping by Multiplexed Shotgun Genotyping (MSG), as well as RNA isolation from single flies. The sample adapter can also hold and shake other items, such as centrifuge tubes (15-50 mL) or small bottles.

  17. High throughput preparation of fly genomic DNA in 96-well format using a paint-shaker

    PubMed Central

    Lang, Michael; Nagy, Olga; Lang, Claus; Orgogozo, Virginie

    2015-01-01

    Sample homogenization is an essential step for genomic DNA extraction, with multiple downstream applications in Molecular Biology. Genotyping hundreds or thousands of samples requires an automation of this homogenization step, and high throughput homogenizer equipment currently costs 7000 euros or more. We present an apparatus for homogenization of individual Drosophila adult flies in 96-well micro-titer dishes, which was built from a small portable paint-shaker (F5 portable paint-shaker, Ushake). Single flies are disrupted in each well that contains extraction buffer and a 4-mm metal ball. Our apparatus can hold up to five 96-well micro-titer plates. Construction of the homogenizer apparatus takes about 3–4 days, and all equipment can be obtained from a home improvement store. The total material cost is approximately 700 euros including the paint-shaker. We tested the performance of our apparatus using the ZR-96 Quick-gDNA™ kit (Zymo Research) homogenization buffer and achieved nearly complete tissue homogenization after 15 minutes of shaking. PCR tests did not detect any cross contamination between samples of neighboring wells. We obtained on average 138 ng of genomic DNA per fly, and DNA quality was adequate for standard PCR applications. In principle, our tissue homogenizer can be used for isolation of DNA suitable for library production and high throughput genotyping by Multiplexed Shotgun Genotyping (MSG), as well as RNA isolation from single flies. The sample adapter can also hold and shake other items, such as centrifuge tubes (15–50 mL) or small bottles. PMID:26818699

  18. A Microfabricated 96-Well 3D Assay Enabling High-Throughput Quantification of Cellular Invasion Capabilities

    PubMed Central

    Hao, Rui; Wei, Yuanchen; Li, Chaobo; Chen, Feng; Chen, Deyong; Zhao, Xiaoting; Luan, Shaoliang; Fan, Beiyuan; Guo, Wei; Wang, Junbo; Chen, Jian

    2017-01-01

    This paper presents a 96-well microfabricated assay to study three-dimensional (3D) invasion of tumor cells. A 3D cluster of tumor cells was first generated within each well by seeding cells onto a micro-patterned surface consisting of a central fibronectin-coated area that promotes cellular attachment, surrounded by a poly ethylene glycol (PEG) coated area that is resistant to cellular attachment. Following the formation of the 3D cell clusters, a 3D collagen extracellular matrix was formed in each well by thermal-triggered gelation. Invasion of the tumor cells into the extracellular matrix was subsequently initiated and monitored. Two modes of cellular infiltration were observed: A549 cells invaded into the extracellular matrix following the surfaces previously coated with PEG molecules in a pseudo-2D manner, while H1299 cells invaded into the extracellular matrix in a truly 3D manner including multiple directions. Based on the processing of 2D microscopic images, a key parameter, namely, equivalent invasion distance (the area of invaded cells divided by the circumference of the initial cell cluster) was obtained to quantify migration capabilities of these two cell types. These results validate the feasibility of the proposed platform, which may function as a high-throughput 3D cellular invasion assay. PMID:28240272

  19. A Microfabricated 96-Well 3D Assay Enabling High-Throughput Quantification of Cellular Invasion Capabilities.

    PubMed

    Hao, Rui; Wei, Yuanchen; Li, Chaobo; Chen, Feng; Chen, Deyong; Zhao, Xiaoting; Luan, Shaoliang; Fan, Beiyuan; Guo, Wei; Wang, Junbo; Chen, Jian

    2017-02-27

    This paper presents a 96-well microfabricated assay to study three-dimensional (3D) invasion of tumor cells. A 3D cluster of tumor cells was first generated within each well by seeding cells onto a micro-patterned surface consisting of a central fibronectin-coated area that promotes cellular attachment, surrounded by a poly ethylene glycol (PEG) coated area that is resistant to cellular attachment. Following the formation of the 3D cell clusters, a 3D collagen extracellular matrix was formed in each well by thermal-triggered gelation. Invasion of the tumor cells into the extracellular matrix was subsequently initiated and monitored. Two modes of cellular infiltration were observed: A549 cells invaded into the extracellular matrix following the surfaces previously coated with PEG molecules in a pseudo-2D manner, while H1299 cells invaded into the extracellular matrix in a truly 3D manner including multiple directions. Based on the processing of 2D microscopic images, a key parameter, namely, equivalent invasion distance (the area of invaded cells divided by the circumference of the initial cell cluster) was obtained to quantify migration capabilities of these two cell types. These results validate the feasibility of the proposed platform, which may function as a high-throughput 3D cellular invasion assay.

  20. Urine sample preparation in 96-well filter plates for quantitative clinical proteomics.

    PubMed

    Yu, Yanbao; Suh, Moo-Jin; Sikorski, Patricia; Kwon, Keehwan; Nelson, Karen E; Pieper, Rembert

    2014-06-03

    Urine is an important, noninvasively collected body fluid source for the diagnosis and prognosis of human diseases. Liquid chromatography mass spectrometry (LC-MS) based shotgun proteomics has evolved as a sensitive and informative technique to discover candidate disease biomarkers from urine specimens. Filter-aided sample preparation (FASP) generates peptide samples from protein mixtures of cell lysate or body fluid origin. Here, we describe a FASP method adapted to 96-well filter plates, named 96FASP. Soluble urine concentrates containing ~10 μg of total protein were processed by 96FASP and LC-MS resulting in 700-900 protein identifications at a 1% false discovery rate (FDR). The experimental repeatability, as assessed by label-free quantification and Pearson correlation analysis for shared proteins among replicates, was high (R ≥ 0.97). Application to urinary pellet lysates which is of particular interest in the context of urinary tract infection analysis was also demonstrated. On average, 1700 proteins (±398) were identified in five experiments. In a pilot study using 96FASP for analysis of eight soluble urine samples, we demonstrated that protein profiles of technical replicates invariably clustered; the protein profiles for distinct urine donors were very different from each other. Robust, highly parallel methods to generate peptide mixtures from urine and other body fluids are critical to increase cost-effectiveness in clinical proteomics projects. This 96FASP method has potential to become a gold standard for high-throughput quantitative clinical proteomics.

  1. Urine Sample Preparation in 96-Well Filter Plates for Quantitative Clinical Proteomics

    PubMed Central

    2015-01-01

    Urine is an important, noninvasively collected body fluid source for the diagnosis and prognosis of human diseases. Liquid chromatography mass spectrometry (LC-MS) based shotgun proteomics has evolved as a sensitive and informative technique to discover candidate disease biomarkers from urine specimens. Filter-aided sample preparation (FASP) generates peptide samples from protein mixtures of cell lysate or body fluid origin. Here, we describe a FASP method adapted to 96-well filter plates, named 96FASP. Soluble urine concentrates containing ∼10 μg of total protein were processed by 96FASP and LC-MS resulting in 700–900 protein identifications at a 1% false discovery rate (FDR). The experimental repeatability, as assessed by label-free quantification and Pearson correlation analysis for shared proteins among replicates, was high (R ≥ 0.97). Application to urinary pellet lysates which is of particular interest in the context of urinary tract infection analysis was also demonstrated. On average, 1700 proteins (±398) were identified in five experiments. In a pilot study using 96FASP for analysis of eight soluble urine samples, we demonstrated that protein profiles of technical replicates invariably clustered; the protein profiles for distinct urine donors were very different from each other. Robust, highly parallel methods to generate peptide mixtures from urine and other body fluids are critical to increase cost-effectiveness in clinical proteomics projects. This 96FASP method has potential to become a gold standard for high-throughput quantitative clinical proteomics. PMID:24797144

  2. Development of a rapid, 96-well alkaline based differential DNA extraction method for sexual assault evidence.

    PubMed

    Hudlow, William R; Buoncristiani, Martin R

    2012-01-01

    We present a rapid alkaline lysis procedure for the extraction of DNA from sexual assault evidence that generates purified sperm fraction extracts that yield STR typing results similar to those obtained from the traditional organic/dithiothreitol differential extraction. Specifically, a sodium hydroxide based differential extraction method has been developed in a single-tube format and further optimized in a 96-well format. The method yields purified extracts from a small sample set (≈ 2-6 swabs) in approximately 2h and from a larger sample set (up to 96 swabs) in approximately 4h. While conventional differential extraction methods require vigorous sample manipulation to remove the spermatozoa from the substrate, the method described here exploits the propensity of sperm to adhere to a substrate and does not require any manipulation of the substrate after it is sampled. For swabs, sample handling is minimized by employing a process where the tip of the swab, including the shaft, is transferred to the appropriate vessel eliminating the need for potentially hazardous scalpels to separate the swab material from the shaft. The absence of multiple handling steps allows the process to be semi-automated, however the procedure as described here does not require use of a robotic system. This method may provide forensic laboratories a cost-effective tool for the eradication of backlogs of sexual assault evidence, and more timely service to their client agencies. In addition, we have demonstrated that a modification of the procedure can be used to retrieve residual sperm-cell DNA from previously extracted swabs.

  3. A human CXCL13-induced actin polymerization assay measured by fluorescence plate reader.

    PubMed

    Alley, Jennifer; Bloom, Laird; Kasaian, Marion; Gao, Huilan; Berstein, Gabriel; Clark, James D; Miao, Wenyan

    2010-02-01

    The chemokine receptor CXCR5 is predominantly expressed on mature B cells and follicular T-helper cells. CXCR5 and its ligand CXCL13 participate in ectopic germinal center formation at the inflammatory sites of multiple immune diseases such as rheumatoid arthritis, multiple sclerosis, and Sjogren's syndrome. Therefore, disrupting CXCL13-induced chemotaxis may be a fruitful approach for developing therapeutics in treating these diseases. Cells undergo cytoskeletal rearrangement prior to chemotaxis, and therefore actin polymerization can be used as a surrogate readout more proximal to chemokine receptor activation than chemotaxis. Conventionally, actin polymerization is measured by fluorescence microscopy or flow cytometry, which are either of low throughput or in need of special instruments. We developed a 96-well actin polymerization assay that can process 1,000 to 1,500 samples a day. This assay uses a standard laboratory fluorescence microplate reader as the detection instrument and was optimized for various experimental conditions such as cell density, actin filament staining reagent, staining buffer, and cell culture conditions. We demonstrate that this actin polymerization assay in 96-well format exhibits the expected pharmacology for human CXCR5 and is suitable as a primary functional assay to screen neutralizing scFv in crude bacterial peri-preps and a secondary assay for small compound collections.

  4. A Quantitative High-Throughput 96-well plate Fluorescence Assay for Mechanism-Based Inactivators of Cytochromes P450 Exemplified using CYP2B6

    PubMed Central

    Kenaan, Cesar; Zhang, Haoming; Hollenberg, Paul F.

    2010-01-01

    Mechanism-based inactivators such as bergamottin are useful chemical tools for identifying the roles of specific active-site amino acid residues in the reactions catalyzed by the cytochromes P450 (CYPs or P450s) that are responsible for the metabolism of a wide variety of drugs and endogenous substrates. In clinical settings mechanism-based inactivation of P450s involved in xenobiotic metabolism has the potential to lead to adverse drug-drug interactions and assays to identify and characterize drug candidates as P450 inactivators are important in drug discovery and development. Here we present a quantitative high-throughput protocol for investigating cytochrome P450 mechanism-based inactivators using the example of CYP2B6 and bergamottin to illustrate the finer points of this protocol. This protocol details the adaptation of a 7-ethoxytrifluoromethyl coumarin (7-EFC) O-deethylation fluorescence activity assay to a 96-well microtiter plate format and uses a plate-reader to detect the fluorescence of the product. Compared to previous methods, this protocol requires less P450 and takes significantly less time while greatly increasing throughput. The protocol as written takes approximately two hours to complete. The principles and procedures outlined in this protocol can be easily adapted to other inactivators, P450 isoforms, substrates and plate-readers. PMID:20885377

  5. General calibration of microbial growth in microplate readers

    PubMed Central

    Stevenson, Keiran; McVey, Alexander F.; Clark, Ivan B. N.; Swain, Peter S.; Pilizota, Teuta

    2016-01-01

    Optical density (OD) measurements of microbial growth are one of the most common techniques used in microbiology, with applications ranging from studies of antibiotic efficacy to investigations of growth under different nutritional or stress environments, to characterization of different mutant strains, including those harbouring synthetic circuits. OD measurements are performed under the assumption that the OD value obtained is proportional to the cell number, i.e. the concentration of the sample. However, the assumption holds true in a limited range of conditions, and calibration techniques that determine that range are currently missing. Here we present a set of calibration procedures and considerations that are necessary to successfully estimate the cell concentration from OD measurements. PMID:27958314

  6. Adjustable bioadhesive control of PEGylated hyperbranch brushes on polystyrene microplate interface for the improved sensitivity of human blood typing.

    PubMed

    Chen, Yan-Wen; Chang, Yung; Lee, Rong-Ho; Li, Wen-Tyng; Chinnathambi, Arunachalam; Alharbi, Sulaiman Ali; Hsiue, Ging-Ho

    2014-08-05

    A PEGylated 96-well polystyrene (PS) microplate was first introduced for applications in high-throughput screening for selective blood typing to minimize the risks in blood transfusions. Herein, we present a hemocompatible PS 96-well microplate with adjustable PEGylated hyperbranch brush coverage prepared by ozone pretreated activation and thermally induced surface PEGylation. The grafting properties, hydration capacity, and blood compatibility of the PEGylated hyperbrush immobilized PS surfaces in human blood were illustrated by the combined chemical and physical properties of the surface, and the dependence of the specific absorption of human plasma fibrinogen onto the PEGylated surfaces on the grafting density was analyzed by monoclonal antibodies. The surface coverage of PEGylated brushes plays a major role in the bioadhesive properties of modified PS microplates, which in turn control the level of agglutination sensitivity in blood typing. The bioadhesive resistance toward proteins, platelets, and erythrocytes in human whole blood showed a correlation to the controlled hydration properties of the PEGylated hyperbrush-modified surfaces. Therefore, we suggested that the surface coverage of PEGylated hyperbrushes on PS surfaces can increase the sensitivity of cross-matching blood agglutination by up to 16-fold compared to that of the conventional 96-well virgin PS due to the regulated biorecognition of hematocrit and antibodies of the PEGylated hyperbrush-modified surfaces.

  7. Quantum dot immunoassays in renewable surface column and 96-well plate formats for the fluorescence detection of Botulinum neurotoxin using high-affinity antibodies

    SciTech Connect

    Warner, Marvin G.; Grate, Jay W.; Tyler, Abby J.; Ozanich, Richard M.; Miller, Keith D.; Lou, Jianlong; Marks, James D.; Bruckner-Lea, Cindy J.

    2009-09-01

    A fluorescence sandwich immunoassay using high affinity antibodies and quantum dot (QD) reporters has been developed for detection of botulinum toxin serotype A (BoNT/A). For the development of the assay, a nontoxic recombinant fragment of the holotoxin (BoNT/A-HC-fragment) has been used as a structurally valid simulant for the full toxin molecule. The antibodies used, AR4 and RAZ1, bind to nonoverlapping epitopes present on both the full toxin and on the recombinant fragment. In one format, the immunoassay is carried out in a 96-well plate with detection in a standard plate reader. Detection down to 31 pM of the BoNT/Hc-fragment was demonstrated with a total incubation time of 3 hours, using AR4 as the capture antibody and QD-coupled RAZ1 as the reporter. In a second format, the AR4 capture antibody was coupled to Sepharose beads, and the immunochemical reactions were carried out in microcentrifuge tubes with an incubation time of 1 hour. These beads were subsequently captured and concentrated in a rotating rod “renewable surface” flow cell as part of a sequential injection fluidic system. This flow cell was equipped with a fiber optic system for fluorescence measurements. In PBS buffer solution matrix, the BoNT/A-HC-fragment was detected to concentrations as low as 5 pM using the fluidic measurement approach.

  8. A semistatic microplate-based phytotoxicity test

    SciTech Connect

    Radetski, C.M.; Ferard, J.F. . Centre des Sciences de l'Environnement); Blaise, C. )

    1995-02-01

    A novel phytotoxicity test is described herein that employs a microplate equipped with membrane-bottomed wells. This MultiScreen[trademark] (Millipore Corp., Bedford, MA) microplate allows performance of a semistatic algal test, in which test medium is renewed periodically. With such a design, the algal test becomes comparable to other short-term tests used to evaluate chronic toxicity of chemicals and effluents. The EC50s obtained for Cu[sup 2+], Cd[sup 2+], Cr[sup 6+], atrazine, and one leachate sample (municipal sludge incinerator residue) with static and semistatic algal microplate tests were compared in this study. The semistatic microplate test revealed greater sensitivity than did the static microplate test.

  9. A 96-well screen filter plate for high-throughput biological sample preparation and LC-MS/MS analysis.

    PubMed

    Peng, Sean X; Cousineau, Martin; Juzwin, Stephen J; Ritchie, David M

    2006-01-01

    A novel 96-well screen filter plate (patent pending) has been invented to eliminate a time-consuming and labor-intensive step in preparation of in vivo study samples--to remove blood or plasma clots. These clots plug the pipet tips during a manual or automated sample-transfer step causing inaccurate pipetting or total pipetting failure. Traditionally, these blood and plasma clots are removed by picking them out manually one by one from each sample tube before any sample transfer can be made. This has significantly slowed the sample preparation process and has become a bottleneck for automated high-throughput sample preparation using robotic liquid handlers. Our novel screen filter plate was developed to solve this problem. The 96-well screen filter plate consists of 96 stainless steel wire-mesh screen tubes connected to the 96 openings of a top plate so that the screen filter plate can be readily inserted into a 96-well sample storage plate. Upon insertion, the blood and plasma clots are excluded from entering the screen tube while clear sample solutions flow freely into it. In this way, sample transfer can be easily completed by either manual or automated pipetting methods. In this report, three structurally diverse compounds were selected to evaluate and validate the use of the screen filter plate. The plasma samples of these compounds were transferred and processed in the presence and absence of the screen filter plate and then analyzed by LC-MS/MS methods. Our results showed a good agreement between the samples prepared with and without the screen filter plate, demonstrating the utility and efficiency of this novel device for preparation of blood and plasma samples. The device is simple, easy to use, and reusable. It can be employed for sample preparation of other biological fluids that contain floating particulates or aggregates.

  10. Ice-Cap: a method for growing Arabidopsis and tomato plants in 96-well plates for high-throughput genotyping.

    PubMed

    Su, Shih-Heng; Clark, Katie A; Gibbs, Nicole M; Bush, Susan M; Krysan, Patrick J

    2011-11-09

    It is becoming common for plant scientists to develop projects that require the genotyping of large numbers of plants. The first step in any genotyping project is to collect a tissue sample from each individual plant. The traditional approach to this task is to sample plants one-at-a-time. If one wishes to genotype hundreds or thousands of individuals, however, using this strategy results in a significant bottleneck in the genotyping pipeline. The Ice-Cap method that we describe here provides a high-throughput solution to this challenge by allowing one scientist to collect tissue from several thousand seedlings in a single day (1,2). This level of throughput is made possible by the fact that tissue is harvested from plants 96-at-a-time, rather than one-at-a-time. The Ice-Cap method provides an integrated platform for performing seedling growth, tissue harvest, and DNA extraction. The basis for Ice-Cap is the growth of seedlings in a stacked pair of 96-well plates. The wells of the upper plate contain plugs of agar growth media on which individual seedlings germinate. The roots grow down through the agar media, exit the upper plate through a hole, and pass into a lower plate containing water. To harvest tissue for DNA extraction, the water in the lower plate containing root tissue is rapidly frozen while the seedlings in the upper plate remain at room temperature. The upper plate is then peeled away from the lower plate, yielding one plate with 96 root tissue samples frozen in ice and one plate with 96 viable seedlings. The technique is named "Ice-Cap" because it uses ice to capture the root tissue. The 96-well plate containing the seedlings can then wrapped in foil and transferred to low temperature. This process suspends further growth of the seedlings, but does not affect their viability. Once genotype analysis has been completed, seedlings with the desired genotype can be transferred from the 96-well plate to soil for further propagation. We have demonstrated

  11. Streptomycetes in micro-cultures: growth, production of secondary metabolites, and storage and retrieval in the 96-well format.

    PubMed

    Minas, W; Bailey, J E; Duetz, W

    2000-12-01

    Mycelium-forming Streptomyces strains were grown in one milliliter liquid micro-cultures in square deep-well microtiter plates. Growth was evaluated with respect to biomass formation and production of secondary metabolites which were found to be very similar in the micro-cultures, bioreactor, and shake flask cultivations, respectively. Despite repetitive sampling and extensive growth on the walls of the wells, no cross contamination occurred. Furthermore, we successfully employed cold storage at -20 degrees C of spore suspensions (in the 96-well format), directly prepared from cultures grown on agar in the microtitre plate. Cultures were retrieved by replicating aliquots from the frozen spore suspensions.

  12. A Novel 96well-formatted Micro-gap Plate Enabling Drug Response Profiling on Primary Tumour Samples

    NASA Astrophysics Data System (ADS)

    Ma, Wei-Yuan; Hsiung, Lo-Chang; Wang, Chen-Ho; Chiang, Chi-Ling; Lin, Ching-Hung; Huang, Chiun-Sheng; Wo, Andrew M.

    2015-04-01

    Drug-based treatments are the most widely used interventions for cancer management. Personalized drug response profiling remains inherently challenging with low cell count harvested from tumour sample. We present a 96well-formatted microfluidic plate with built-in micro-gap that preserves up to 99.2% of cells during multiple assay/wash operation and only 9,000 cells needed for a single reagent test (i.e. 1,000 cells per test spot x 3 selected concentration x triplication), enabling drug screening and compatibility with conventional automated workstations. Results with MCF7 and MDA-MB-231 cell lines showed that no statistical significance was found in dose-response between the device and conventional 96-well plate control. Primary tumour samples from breast cancer patients tested in the device also showed good IC50 prediction. With drug screening of primary cancer cells must consider a wide range of scenarios, e.g. suspended/attached cell types and rare/abundant cell availability, the device enables high throughput screening even for suspended cells with low cell count since the signature microfluidic cell-trapping feature ensures cell preservation in a multiple solution exchange protocol.

  13. Protocol: high throughput silica-based purification of RNA from Arabidopsis seedlings in a 96-well format

    PubMed Central

    2011-01-01

    The increasing popularity of systems-based approaches to plant research has resulted in a demand for high throughput (HTP) methods to be developed. RNA extraction from multiple samples in an experiment is a significant bottleneck in performing systems-level genomic studies. Therefore we have established a high throughput method of RNA extraction from Arabidopsis thaliana to facilitate gene expression studies in this widely used plant model. We present optimised manual and automated protocols for the extraction of total RNA from 9-day-old Arabidopsis seedlings in a 96 well plate format using silica membrane-based methodology. Consistent and reproducible yields of high quality RNA are isolated averaging 8.9 μg total RNA per sample (~20 mg plant tissue). The purified RNA is suitable for subsequent qPCR analysis of the expression of over 500 genes in triplicate from each sample. Using the automated procedure, 192 samples (2 × 96 well plates) can easily be fully processed (samples homogenised, RNA purified and quantified) in less than half a day. Additionally we demonstrate that plant samples can be stored in RNAlater at -20°C (but not 4°C) for 10 months prior to extraction with no significant effect on RNA yield or quality. Additionally, disrupted samples can be stored in the lysis buffer at -20°C for at least 6 months prior to completion of the extraction procedure providing a flexible sampling and storage scheme to facilitate complex time series experiments. PMID:22136293

  14. Mass Spectrometric N-Glycan Analysis of Haptoglobin from Patient Serum Samples Using a 96-Well Plate Format.

    PubMed

    Zhu, Jianhui; Wu, Jing; Yin, Haidi; Marrero, Jorge; Lubman, David M

    2015-11-06

    Alterations in glycosylation of serum glycoproteins can provide unique and highly specific fingerprints of malignancy. Our previous mass spectrometric study revealed that the bifucosylation level of serum haptoglobin was distinctly increased in hepatocellular carcinoma (HCC) patients versus liver cirrhosis of all three major etiologies. We have thus developed a method for the analysis of large numbers of serum samples based on a 96-well plate platform for the evaluation of fucosylation changes of serum haptoglobin between HCC versus cirrhosis. Haptoglobin was isolated from the serum of individual patient samples based on an HPLC column immobilized with antihaptoglobin antibody via hydrazide immobilization chemistry. Only 10 μL of serum was required for glycan extraction and processing for MALDI-QIT mass spectrometry analysis using the 96-well plate format. The bifucosylation degrees of haptoglobin in individuals were calculated using a quantitative glycomics method. The MS data confirmed that the bifucosylated tetra-anntenary glycan was upregulated in HCC samples of all etiologies. This study provides a parallel method for processing glycan content for haptoglobin and evaluating detailed changes in glycan structures for a potentially large cohort of clinical serum samples.

  15. Seismicity of the Adriatic microplate

    USGS Publications Warehouse

    Console, R.; Di, Giovambattista R.; Favali, P.; Presgrave, B.W.; Smriglio, G.

    1993-01-01

    The Adriatic microplate was previously considered to be a unique block, tectonically active only along its margins. The seismic sequences that took place in the basin from 1986 to 1990 give new information about the geodynamics of this area. Three subsets of well recorded events were relocated by the joint hypocentre determination technique. On the whole, this seismic activity was concentrated in a belt crossing the southern Adriatic sea around latitude 42??, in connection with regional E-W fault systems. Some features of this seismicity, similar to those observed in other well known active margins of the Adriatic plate, support a model of a southern Adriatic lithospheric block, detached from the Northern one. Other geophysical information provides evidence of a transitional zone at the same latitude. ?? 1993.

  16. Performance evaluation of 3D polystyrene 96-well plates with human neural stem cells in a calcium assay.

    PubMed

    Lai, Yinzhi; Kisaalita, William S

    2012-08-01

    In this study, we have generated a high-throughput screening (HTS)-compatible 3D cell culture platform by chemically "welding" polystyrene scaffolds into standard 2D polystyrene 96-well plates. The variability of scaffolds was minimized by introducing automation into the fabrication process. The fabricated 3D cell culture plates were compared with several commercially available 3D cell culture platforms with light and scanning electron microscopy. Voltage-gated calcium channel functionality was used to access the Z' factors of all plates, including a 2D standard plate control. It was found that with the No-Wash Fluo-4 calcium assay and neural progenitor cells, all plates display acceptable Z' factors for use in HTS. The plates with "welded" polystyrene scaffolds have several advantages, such as being versatile and economical, and are ready to use off the shelf. These characteristics are especially desired in HTS preclinical drug discovery applications.

  17. Development and validation of a 96-well cellular assay for the discovery of ALDH1A1 inhibitors.

    PubMed

    Ming, Wenyu; Ma, Wenzhen; Chen, Lisa H; Volk, Catherine; Michael, Mervyn Dodson; Xu, Yanping; Zhang, Fang; Wang, Xiaojun

    2013-07-01

    Retinoic acid, the active metabolite of vitamin A, plays important roles in various physiological and pathological processes. The two-step production of retinoic acid from vitamin A (retinol) is catalyzed by alcohol dehydrogenases and aldehyde dehydrogenases, which are potential therapeutic targets for numerous diseases, such as obesity, diabetes, and cancer. Currently, the lack of a suitable high-throughput cellular assay hinders efforts to identify therapeutic small molecular inhibitors of aldehyde dehydrogenase, such as ALDH1A1. In this report, we utilized high-content imaging technology and a commercially available cell permeable ALDH substrate to develop a 96-well cellular ALDH1A1 assay. This assay has a robust and sensitive readout and is amenable to automation. With this cellular assay, we identified potent selective ALDH1A1 inhibitors to explore the role of retinoic acid production in various preclinical disease models.

  18. 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.

  19. Scalable 96-well Plate Based iPSC Culture and Production Using a Robotic Liquid Handling System.

    PubMed

    Conway, Michael K; Gerger, Michael J; Balay, Erin E; O'Connell, Rachel; Hanson, Seth; Daily, Neil J; Wakatsuki, Tetsuro

    2015-05-14

    Continued advancement in pluripotent stem cell culture is closing the gap between bench and bedside for using these cells in regenerative medicine, drug discovery and safety testing. In order to produce stem cell derived biopharmaceutics and cells for tissue engineering and transplantation, a cost-effective cell-manufacturing technology is essential. Maintenance of pluripotency and stable performance of cells in downstream applications (e.g., cell differentiation) over time is paramount to large scale cell production. Yet that can be difficult to achieve especially if cells are cultured manually where the operator can introduce significant variability as well as be prohibitively expensive to scale-up. To enable high-throughput, large-scale stem cell production and remove operator influence novel stem cell culture protocols using a bench-top multi-channel liquid handling robot were developed that require minimal technician involvement or experience. With these protocols human induced pluripotent stem cells (iPSCs) were cultured in feeder-free conditions directly from a frozen stock and maintained in 96-well plates. Depending on cell line and desired scale-up rate, the operator can easily determine when to passage based on a series of images showing the optimal colony densities for splitting. Then the necessary reagents are prepared to perform a colony split to new plates without a centrifugation step. After 20 passages (~3 months), two iPSC lines maintained stable karyotypes, expressed stem cell markers, and differentiated into cardiomyocytes with high efficiency. The system can perform subsequent high-throughput screening of new differentiation protocols or genetic manipulation designed for 96-well plates. This technology will reduce the labor and technical burden to produce large numbers of identical stem cells for a myriad of applications.

  20. Scalable 96-well Plate Based iPSC Culture and Production Using a Robotic Liquid Handling System

    PubMed Central

    Conway, Michael K.; Gerger, Michael J.; Balay, Erin E.; O'Connell, Rachel; Hanson, Seth; Daily, Neil J.; Wakatsuki, Tetsuro

    2015-01-01

    Continued advancement in pluripotent stem cell culture is closing the gap between bench and bedside for using these cells in regenerative medicine, drug discovery and safety testing. In order to produce stem cell derived biopharmaceutics and cells for tissue engineering and transplantation, a cost-effective cell-manufacturing technology is essential. Maintenance of pluripotency and stable performance of cells in downstream applications (e.g., cell differentiation) over time is paramount to large scale cell production. Yet that can be difficult to achieve especially if cells are cultured manually where the operator can introduce significant variability as well as be prohibitively expensive to scale-up. To enable high-throughput, large-scale stem cell production and remove operator influence novel stem cell culture protocols using a bench-top multi-channel liquid handling robot were developed that require minimal technician involvement or experience. With these protocols human induced pluripotent stem cells (iPSCs) were cultured in feeder-free conditions directly from a frozen stock and maintained in 96-well plates. Depending on cell line and desired scale-up rate, the operator can easily determine when to passage based on a series of images showing the optimal colony densities for splitting. Then the necessary reagents are prepared to perform a colony split to new plates without a centrifugation step. After 20 passages (~3 months), two iPSC lines maintained stable karyotypes, expressed stem cell markers, and differentiated into cardiomyocytes with high efficiency. The system can perform subsequent high-throughput screening of new differentiation protocols or genetic manipulation designed for 96-well plates. This technology will reduce the labor and technical burden to produce large numbers of identical stem cells for a myriad of applications. PMID:26068617

  1. 96-Well Plate Colorimetric Assay for K(sub i) Determination of (plusmn)-2-Benzylsuccinic Acid, an Inhibitor of Carboxypeptidase A

    ERIC Educational Resources Information Center

    Wentland, Mark P.; Raza, Shaan; Yingtong Gao

    2004-01-01

    An appropriate assay to determine the inhibition potency of carboxypeptidase A (CPA) in 96-well format to illustrate how high throughput screening is used in modern drug discovery to identify bioactive molecules is developed. Efforts in developing a colorimetric 96-well plate assay for determination of the K(sub i) for inhibition of CPA by…

  2. Microplate fecal coliform method to monitor stream water pollution.

    PubMed Central

    Maul, A; Block, J C

    1983-01-01

    A study has been carried out on the Moselle River by means of a microtechnique based on the most-probable-number method for fecal coliform enumeration. This microtechnique, in which each serial dilution of a sample is inoculated into all 96 wells of a microplate, was compared with the standard membrane filter method. It showed a marked overestimation of about 14% due, probably, to the lack of absolute specificity of the method. The high precision of the microtechnique (13%, in terms of the coefficient of variation for log most probable number) and its relative independence from the influence of bacterial density allowed the use of analysis of variance to investigate the effects of spatial and temporal bacterial heterogeneity on the estimation of coliforms. Variability among replicate samples, subsamples, handling, and analytical errors were considered as the major sources of variation in bacterial titration. Variances associated with individual components of the sampling procedure were isolated, and optimal replications of each step were determined. Temporal variation was shown to be more influential than the other three components (most probable number, subsample, sample to sample), which were approximately equal in effect. However, the incidence of sample-to-sample variability (16%, in terms of the coefficient of variation for log most probable number) caused by spatial heterogeneity of bacterial populations in the Moselle River is shown and emphasized. Consequently, we recommend that replicate samples be taken on each occasion when conducting a sampling program for a stream pollution survey. PMID:6360044

  3. Lipophilic rather than hydrophilic photosensitizers show strong adherence to standard cell culture microplates under cell-free conditions.

    PubMed

    Engelhardt, Victoria; Kiesslich, Tobias; Berlanda, Juergen; Hofbauer, Stefanie; Krammer, Barbara; Plaetzer, Kristjan

    2011-06-02

    Analysis of photosensitizer (PS) uptake kinetics into tumor cells is a standard cell culture experiment in photodynamic therapy (PDT) - usually performed in plastic microplates or petri dishes. Organic substances such as PS can potentially interact with the plastic surfaces. In this study, we provide a qualitative comparison of three lipophilic PS (hypericin, Foscan® and Photofrin®) and two rather hydrophilic PS formulations (PVP-hypericin and aluminum (III) phthalocyanine tetrasulfonate chloride) regarding their adherence to the surfaces of 96-well microplates obtained from four different manufacturers. For estimation of the relevance of PS adherence for cellular uptake studies we compared the fluorescence signal of the respective PS in microplates containing A431 human epithelial carcinoma cells with microplates incubated with the respective PS under cell-free conditions. We demonstrate that lipophilic PS substances show a strong adherence to microplates - in case of direct lysis and fluorescence measurement resulting in 50% up to 90% of the overall signal to be caused by adherence of the substances to the plastic materials in a cellular uptake experiment. For the hydrophilic compounds, adherence is negligible. Interestingly, adherence of PS agents to microplates takes place in a time-dependent and thus kinetic-like manner, requiring up to several hours to reach a plateau of the fluorescence signal. Furthermore, PS adherence is a function of the PS concentration applied and no saturation effect was observed for the concentrations used in this study. Taken together, this study provides a systematic analysis under which conditions PS adherence to cell culture plates may contribute to the overall fluorescence signal in - for example - PS uptake experiments.

  4. Equilibrium drug solubility measurements in 96-well plates reveal similar drug solubilities in phosphate buffer pH 6.8 and human intestinal fluid.

    PubMed

    Heikkilä, Tiina; Karjalainen, Milja; Ojala, Krista; Partola, Kirsi; Lammert, Frank; Augustijns, Patrick; Urtti, Arto; Yliperttula, Marjo; Peltonen, Leena; Hirvonen, Jouni

    2011-02-28

    This study was conducted to develop a high throughput screening (HTS) method for the assessment of equilibrium solubility of drugs. Solid-state compounds were precipitated from methanol in 96-well plates, in order to eliminate the effect of co-solvent. Solubility of twenty model drugs was analyzed in water and aqueous solutions (pH 1.2 and 6.8) in 96-well plates and in shake-flasks (UV detection). The results obtained with the 96-well plate method correlated well (R(2)=0.93) between the shake-flask and 96-well plates over the wide concentration scale of 0.002-169.2mg/ml. Thereafter, the solubility tests in 96-well plates were performed using fasted state human intestinal fluid (HIF) from duodenum of healthy volunteers. The values of solubility were similar in phosphate buffer solution (pH 6.8) and HIF over the solubility range of 10(2)-10(5)μg/ml. The new 96-well plate method is useful for the screening of equilibrium drug solubility during the drug discovery process and it also allows the use of human intestinal fluid in solubility screening.

  5. Assessment of a 96-Well Plate Assay of Quantitative Drug Susceptibility Testing for Mycobacterium Tuberculosis Complex in China

    PubMed Central

    Xia, Hui; Zheng, Yang; Zhao, Bing; van den Hof, Susan; Cobelens, Frank; Zhao, YanLin

    2017-01-01

    Objective To evaluate the performance of the Sensitire MYCOTB MIC Plate (MYCOTB) which could measure the twelve anti-tuberculosis drugs susceptibility on one 96-wells plate. Methods A total of 140 MDR-TB strains and 60 non-MDR strains were sub-cultured and 193 strains were finally tested for drug resistance using MYCOTB and agar proportion method (APM) and another 7 strains failed of subculture. The drugs included ofloxacin (Ofx), moxifloxacin (Mfx), rifampin (RFP), amikacin (Am), rifabutin (Rfb), para-aminosalicylic acid (PAS), ethionamide (Eth), isoniazid (INH), kanamycin (Km), ethambutol (EMB), streptomycin (Sm), and cycloserine(Cs). The categorical agreement, conditional agreement, sensitivity and specificity of MYCOTB were assessed in comparison with APM. For strains with inconsistent results between MYCOTB and APM, the drug resistance related gene fragments were amplified and sequenced: gyrA for Ofx and Mfx; rpoB for RFP and Rfb; embB for EMB; rpsl for Sm; katG and the promoter region of inhA for INH, ethA and the promoter region of inhA for Eth. The sequence results were compared with results of MYCOTB and APM to analyze the consistency between sequence results and MYCOTB or APM. Results The categorical agreement between two methods for each drug ranged from 88.6% to 100%. It was the lowest for INH (88.6%). The sensitivity and specificity of MYCOTB ranged from 71.4% to 100% and 84.3% to 100%, respectively. The sensitivity was lowest for Cs(71.4%), EMB at 10μg/ml (80.0%) and INH at 10.0μg/ml (84.6%). The specificity was lowest for Rfb (84.3%). Overall discordance between the two phenotypic methods was observed for 96 strains, of which 63 (65.6%) were found susceptible with APM and resistant with MYCOTB and the remaining 33(34.4%) strains were resistant by APM and susceptible with MYCOTB. 34/52 (65.4%) sequenced APM susceptible and MYCOTB resistant(APM-S/MYCOTB-R) strains had mutations or insertions in the amplified regions. 20/30 (66.7%) sequenced APM

  6. Computer-assisted photometric microplate analysis.

    PubMed

    Hörer, O L; Pop, D A

    1987-01-01

    The main algorithm of computer-assisted absorption and emission photometry of samples on a microplate is presented. The software can be used for the enzyme immunoassay (EIA) and other virological tests. The performances of an SPF-500 (Aminco) spectrofluorometer/Felix M18 microcomputer system are discussed on the ground of some results obtained by using the implemented programs.

  7. Lifelong Readers.

    ERIC Educational Resources Information Center

    Sorba, Barbara

    A brief review of the literature on the various approaches which have been used to help children formulate their emerging attitudes towards reading and literature helps clarify the goal of creating lifelong readers. Samway (1991), discussing literature study circles, relates that students read the book they have selected, answer assigned…

  8. COCHABAMBA READER.

    ERIC Educational Resources Information Center

    LASTRA, YOLANDA

    INTENDED AS FOLLOWUP MATERIAL AFTER THE COMPLETION OF THE TWO-VOLUME SPOKEN COCHABAMBA COURSE, THIS READER CONSISTS OF A SINGLE LONG STORY, "JUANITO," WRITTEN BY OSCAR TERAN. IT HAS BEEN USED AS A RADIO SCRIPT FOR A SERIES OF BROADCASTS FROM A COCHABAMBA STATION WHICH SERVES THE SURROUNDING INDIGENOUS POPULATION. THE MATERIAL IS…

  9. Highly Multiplexed RNA Aptamer Selection using a Microplate-based Microcolumn Device

    PubMed Central

    Reinholt, Sarah J.; Ozer, Abdullah; Lis, John T.; Craighead, Harold G.

    2016-01-01

    We describe a multiplexed RNA aptamer selection to 19 different targets simultaneously using a microcolumn-based device, MEDUSA (Microplate-based Enrichment Device Used for the Selection of Aptamers), as well as a modified selection process, that significantly reduce the time and reagents needed for selections. We exploited MEDUSA’s reconfigurable design between parallel and serially-connected microcolumns to enable the use of just 2 aliquots of starting library, and its 96-well microplate compatibility to enable the continued use of high-throughput techniques in downstream processes. Our modified selection protocol allowed us to perform the equivalent of a 10-cycle selection in the time it takes for 4 traditional selection cycles. Several aptamers were discovered with nanomolar dissociation constants. Furthermore, aptamers were identified that not only bound with high affinity, but also acted as inhibitors to significantly reduce the activity of their target protein, mouse decapping exoribonuclease (DXO). The aptamers resisted DXO’s exoribonuclease activity, and in studies monitoring DXO’s degradation of a 30-nucleotide substrate, less than 1 μM of aptamer demonstrated significant inhibition of DXO activity. This aptamer selection method using MEDUSA helps to overcome some of the major challenges with traditional aptamer selections, and provides a platform for high-throughput selections that lends itself to process automation. PMID:27432610

  10. A simple and rapid microplate assay for glycoprotein-processing glycosidases.

    PubMed

    Kang, M S; Zwolshen, J H; Harry, B S; Sunkara, P S

    1989-08-15

    A simple and convenient microplate assay for glycosidases involved in the glycoprotein-processing reactions is described. The assay is based on specific binding of high-mannose-type oligosaccharide substrates to concanavalin A-Sepharose, while monosaccharides liberated by enzymatic hydrolysis do not bind to concanavalin A-Sepharose. By the use of radiolabeled substrates [( 3H]glucose for glucosidases and [3H]mannose for mannosidases), the radioactivity in the liberated monosaccharides can be determined as a measure of the enzymatic activity. This principle was employed earlier for developing assays for glycosidases previously reported (B. Saunier et al. (1982) J. Biol. Chem. 257, 14155-14161; T. Szumilo and A. D. Elbein (1985) Anal. Biochem. 151, 32-40). These authors have reported the separation of substrate from the product by concanavalin A-Sepharose column chromatography. This procedure is handicapped by the fact that it cannot be used for a large number of samples and is time consuming. We have simplified this procedure and adapted it to the use of a microplate (96-well plate). This would help in processing a large number of samples in a short time. In this report we show that the assay is comparable to the column assay previously reported. It is linear with time and enzyme concentration and shows expected kinetics with castanospermine, a known inhibitor of alpha-glucosidase I.

  11. A simple and rapid microplate assay for glycoprotein-processing glycosidases

    SciTech Connect

    Kang, M.S.; Zwolshen, J.H.; Harry, B.S.; Sunkara, P.S. )

    1989-08-15

    A simple and convenient microplate assay for glycosidases involved in the glycoprotein-processing reactions is described. The assay is based on specific binding of high-mannose-type oligosaccharide substrates to concanavalin A-Sepharose, while monosaccharides liberated by enzymatic hydrolysis do not bind to concanavalin A-Sepharose. By the use of radiolabeled substrates (( 3H)glucose for glucosidases and (3H)mannose for mannosidases), the radioactivity in the liberated monosaccharides can be determined as a measure of the enzymatic activity. This principle was employed earlier for developing assays for glycosidases previously reported. These authors have reported the separation of substrate from the product by concanavalin A-Sepharose column chromatography. This procedure is handicapped by the fact that it cannot be used for a large number of samples and is time consuming. We have simplified this procedure and adapted it to the use of a microplate (96-well plate). This would help in processing a large number of samples in a short time. In this report we show that the assay is comparable to the column assay previously reported. It is linear with time and enzyme concentration and shows expected kinetics with castanospermine, a known inhibitor of alpha-glucosidase I.

  12. Versatile wetting measurement of microplate wells

    NASA Astrophysics Data System (ADS)

    Ng, Enoch Ming Wei; Cheong, Brandon Huey-Ping; Yu, Yang; Liew, Oi Wah; Ng, Tuck Wah

    2016-11-01

    A method to measure the contact angle, which is indicative of wetting, using small liquid volumes dispensed directly on microplate wells is described and demonstrated. Experiments with enhanced green protein samples of volumes 4.4-6 μl showed no measured variance in the contact angle. Experiments with phosphate buffer solution with varied concentrations of a non-ionic detergent (Tween 20) dissolved, however, revealed smaller contact angles with increased detergent concentration. It is experimentally shown that drops can be located up to 7° from the lowest position of the well without affecting the accuracy of contact angle measurements. Numerical simulations confirm the ability of the drops to manifest the correct contact angle despite the lack of axis-symmetry in their shape while residing on a circular surface. This method offers a convenient means to determine the wetting characteristics of different liquid samples in different microplates.

  13. In vitro toxicity testing with microplate cell cultures: Impact of cell binding.

    PubMed

    Gülden, Michael; Schreiner, Jeannine; Seibert, Hasso

    2015-06-05

    In vitro generated data on toxic potencies are generally based on nominal concentrations. However, cellular and extracellular binding and elimination processes may reduce the available free fraction of a compound. Then, nominal effective concentrations do not represent appropriate measures of toxic exposure in vitro and underestimate toxic potencies. In this study it was investigated whether cell binding can affect the availability of chemicals in microplate based toxicity assays. To this end the cytotoxicity of compounds like mercury chloride, digitonin and alcohol ethoxylates, accumulated by cells via different modes, was investigated in 96-well microplate cultures with varying concentrations of Balb/c 3T3 cells. The median effective nominal concentrations of all but one of the tested compounds depended linearly from the cell concentration. Applying a previously developed equilibrium distribution model cell concentration-independent median effective extracellular concentrations and cell burdens, respectively, could be calculated. The compounds were accumulated by the cells with bioconcentration factors, BCF, between 480 and ≥ 25,000. Cell binding of the alcohol ethoxylates was correlated with their lipophilicity. The results show that significant cell binding can occur even at the small cell volume fractions (∼ 1 × 10(-5) to 3 × 10(-3) L/L) encountered in microplate assays. To what extent cell binding affects the bioavailability depends on the BCF and the cell volume fraction. EC50 measurements in the presence of at least two different cell concentrations allow for excluding or detecting significant cell binding and for determining more appropriate measures of toxic exposure in vitro like median effective extracellular (free) concentrations or cell burdens.

  14. Microplate quantification of enzymes of the plant ascorbate-glutathione cycle.

    PubMed

    Murshed, Ramzi; Lopez-Lauri, Félicie; Sallanon, Huguette

    2008-12-15

    Here, we describe microplate assays for determining the specific activities of four enzymes that constitute the ascorbate-glutathione cycle: APX, MDHAR, DHAR, and GR. In plants, these enzymes play a major role in detoxifying reactive oxygen species produced in cells under environmental stress. This work presents the development of plate reader assays to allow rapid analysis of the ascorbate-glutathione cycle activity using tomato fruits subjected to salt stress as a model. With this method, it is possible to analyze easily in one day the activities of the four enzymes for 30 experimental samples, all in triplicate and with blanks.

  15. A new approach to the application of solid phase extraction disks with LC-MS/MS for the analysis of drugs on a 96-well plate format.

    PubMed

    Cudjoe, Erasmus; Pawliszyn, Janusz

    2009-11-01

    A new 96-well disk solid phase extraction sample preparation technique which does not involve vacuum pumps integrated with liquid chromatographic tandem mass spectrometric (LC-MS/MS) was developed for high throughput determination of benzodiazepines (nordiazepam, diazepam, lorazepam and oxazepam). In addition, the method completely allows the re-use of the SPE disk membranes for subsequent analyses after re-conditioning. The method utilizes a robotic autosampler for parallel extractions in a 96-well plate format. Results have been presented for independent extractions from three matrices; phosphate buffer solution, urine, and plasma. Factors affecting data reproducibility, extraction kinetics, sample throughput, and reliability of the system were investigated and optimized. A total time required per sample was 0.94 min using 96-well format. Method reproducibility was < or =9% relative standard deviation for all three matrices. Limits of detection and quantitation recorded were respectively in the range 0.02-0.15 and 0.2-2.0 ng/mL with linearity ranging from 0.2 to 500 ng/mL for all matrices.

  16. Helium isotope ratios in Easter microplate basalts

    NASA Astrophysics Data System (ADS)

    Poreda, R. J.; Schilling, J. G.; Craig, H.

    1993-09-01

    He-3/He-4 ratios in Easter Microplate basalt glasses show clear evidence of the effects of a mantle plume. The East Rift of the microplate between 26 and 28 deg S, identified by La/Sm, Sr and Pb isotopes and ridge crest elevation as the region of maximum plume influence, has He-3/He-4 ratios spanning the entire range from 7.5 to 11.7 R(sub A). The Easter Microplate is the only section of the entire East Pacific Rise that is associated with a known `hotspot' track (mantle plume) and has elevated He-3/He-4 ratios. Although most of the West Rift basalts contain MORB helium (8.0 - 8.7 (R sub A)), the basalt closest to the East Rift has an elevated He-3/He-4 ratio (11.3 R(sub A)), consistent with a significant plume component. The diversity in isotopic signatures also indicates that homogenization of isotopic anomalies does not occur, even in this region of `super-fast' spreading. The overall He-3/He-4-Pb-206/Pb-204 and He-3/He-4-Sr-87/Sr-86 trends have positive correlations, although the high between the He and Sr isotope distribution is modeled in the context of a plume source-migrating ridge sink. During channeling of the plume toward the ridge, helium if preferentially lost from the center of the channeled plume, resulting in lower He/Pb and He/Sr concentration ratios in the high He-3/He-4 component. Mixing trajectories in He-Sr isotopic space between a LILE depleted asthenosphere and a variably degassed plume component provide a reasonably good fit to the data and may explain the isotope systematics of plume-ridge interactions in the context of modern theories of plume dynamics.

  17. Portable Microfiche Readers.

    ERIC Educational Resources Information Center

    Zybura, Edward L.

    1980-01-01

    Provides an introduction to use of three types of portable microfiche readers: hand-held readers, briefcase readers, and self-contained mobile readers. Features described include image quality, method of operation, product history, and optimal environments for utilization. (SW)

  18. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates

    PubMed Central

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-01-01

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format. PMID:27886235

  19. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates

    NASA Astrophysics Data System (ADS)

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-11-01

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format.

  20. 96-well plate-to-plate gravity fluorous solid-phase extraction (F-SPE) for solution-phase library purification.

    PubMed

    Zhang, Wei; Lu, Yimin

    2007-01-01

    Large particle size (125_210 microm) fluorous silica gel bonded with a -SiCH2CH2C8F17 stationary phase has been employed for gravity-driven fluorous solid-phase extraction (F-SPE) on two types of 96-well plates. A 1 or 0.75 g portion of fluorous silica is packed to each well of the 3.5-mL Ex-Blok and the 2.2-mL deep-well filtration plates, respectively. Up to 50 mg of reaction mixture is loaded and then eluted with a fluorophobic solvent (DMSO, DMF, or 85:15 DMF-H2O). Products collected in 96-well receiving plates are directly concentrated on a GeneVac vacuum centrifuge. This simple and highly efficient plate-to-plate F-SPE technique has been demonstrated in the purification of four 96-compound libraries produced by scavenging reactions with 1-(perfluoroctyl)propyl isatoic anhydride (F-IA), amide coupling reactions with 2-chloro-4,6-bis[(perfluorooctyl)propyloxy]-1,3,5-triazine (F-CDMT) or 2,4-dichloro-6-(perfluorooctyl)propyloxy-1,3,5-triazine (F-DCT), and Mitsunobu reactions with fluorous diethyl azodicarboxylate (F-DEAD) and triphenylphosphine (F-TPP). Approximately 80% of products in each library have greater than 85% purity after F-SPE without conducting chromatography.

  1. Higher throughput bioanalysis by automation of a protein precipitation assay using a 96-well format with detection by LC-MS/MS.

    PubMed

    Watt, A P; Morrison, D; Locker, K L; Evans, D C

    2000-03-01

    Generic methodology for the automated preparation and analysis of drug levels in plasma samples within a drug discovery environment was achieved through the redesign of a protein precipitation assay to a microtiter (96-well) plate format and the application of robotic liquid handling for performance of all transfer and pipetting steps. Validation studies revealed that the application of robotics to sample preparation, in general, maintained the analytical accuracy and precision compared with preparing samples manually. The use of rapid gradient LC-MS/MS for analysis coupled with flow diversion of the solvent front allowed the introduction of protein-precipitated samples into the mass spectrometer without the necessity for source cleaning. The problem inherent in automatically pipetting plasma, caused by fibrinogen clots, was overcome by storing samples at -80 degrees C and thus precluding clot formation. The resulting methodology allowed sample preparation for a 96-well plate designed to accommodate 54 unknowns, duplicate 12-point calibration curves, and 6 sets of quality controls at three levels in approximately 2 h. This approach allowed an increase in throughput of sample preparation and analysis to >400 samples per day per LC-MS/MS instrument with minimal manual intervention. Overall, substantial time savings were realized, demonstrating that automation is an increasingly essential tool in a drug discovery bioanalytical environment.

  2. High-throughput 96-well solvent mediated sonic blending synthesis and on-plate solid/solution stability characterization of pharmaceutical cocrystals.

    PubMed

    Luu, Van; Jona, Janan; Stanton, Mary K; Peterson, Matthew L; Morrison, Henry G; Nagapudi, Karthik; Tan, Helming

    2013-01-30

    A 96-well high-throughput cocrystal screening workflow has been developed consisting of solvent-mediated sonic blending synthesis and on-plate solid/solution stability characterization by XRPD. A strategy of cocrystallization screening in selected blend solvents including water mixtures is proposed to not only manipulate solubility of the cocrystal components but also differentiate physical stability of the cocrystal products. Caffeine-oxalic acid and theophylline-oxalic acid cocrystals were prepared and evaluated in relation to saturation levels of the cocrystal components and stability of the cocrystal products in anhydrous and hydrous solvents. AMG 517 was screened with a number of coformers, and solid/solution stability of the resulting cocrystals on the 96-well plate was investigated. A stability trend was observed and confirmed that cocrystals comprised of lower aqueous solubility coformers tended to be more stable in water. Furthermore, cocrystals which could be isolated under hydrous solvent blending condition exhibited superior physical stability to those which could only be obtained under anhydrous condition. This integrated HTS workflow provides an efficient route in an API-sparing approach to screen and identify cocrystal candidates with proper solubility and solid/solution stability properties.

  3. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates.

    PubMed

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-11-25

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format.

  4. India-Eurasia collision triggers formation of an oceanic microplate

    NASA Astrophysics Data System (ADS)

    Matthews, Kara; Müller, Dietmar; Sandwell, David

    2016-04-01

    Detailed mapping of seafloor tectonic fabric in the Indian Ocean, using high-resolution satellite-derived vertical gravity gradient data, reveals an extinct Pacific-style oceanic microplate - the Mammerickx Microplate - west of the Ninetyeast Ridge. It is one of the first Pacific-style microplates to be mapped outside the Pacific basin, suggesting that geophysical conditions during formation probably resembled those that have dominated at eastern Pacific ridges. The microplate formed at the Indian-Antarctic ridge and is bordered by an extinct ridge in the north and pseudofault in the south, whose conjugate is located north of the Kerguelen Plateau. Independent microplate rotation is indicated by asymmetric pseudofaults and rotated abyssal hill fabric, also identified in multibeam data. Magnetic anomaly picks and age estimates calculated from published spreading rates suggest formation during chron 21o (~47.3 Ma). Plate reorganizations can trigger ridge propagation and microplate development, and we propose that formation of the Mammerickx Microplate is linked with the initial 'soft' stage of the India-Eurasia collision. The collision altered the stress regime at the Indian-Antarctic ridge, leading to a change in segmentation and ridge propagation from an establishing transform fault. Fast Indian-Antarctic spreading that preceded microplate formation, and Kerguelen Plume activity may have facilitated ridge propagation via the production of thin and weak lithosphere. However, both factors had been present for tens of millions of years and are therefore unlikely to have triggered the event. Prior to the collision, this combination of fast spreading and plume activity was responsible for the production of a wide region of undulate seafloor to the north of the extinct ridge and 'W' shaped lineations that record back and forth ridge propagation. Microplate formation provides a means of dating the onset of the India-Eurasia collision, and is completely independent of and

  5. Rapid Screening of the Epidermal Growth Factor Receptor Phosphosignaling Pathway via Microplate-Based Dot Blot Assays

    PubMed Central

    Cappione, Amedeo; Smith, Janet; Mabuchi, Masaharu; Nadler, Timothy

    2012-01-01

    Expression profiling on a large scale, as is the case in drug discovery, is often accomplished through use of sophisticated solid-phase protein microarrays or multiplex bead technologies. While offering both high-throughput and high-content analysis, these platforms are often too cost prohibitive or technically challenging for many research settings. Capitalizing on the favorable attributes of the standard ELISA and slot blotting techniques, we developed a modified dot blot assay that provides a simple cost-effective alternative for semiquantitative expression analysis of multiple proteins across multiple samples. Similar in protocol to an ELISA, but based in a membrane bound 96-well microplate, the assay takes advantage of vacuum filtration to expedite the tedious process of washing in between binding steps. We report on the optimization of the assay and demonstrate its use in profiling temporal changes in phosphorylation events in the well-characterized EGF-induced signaling cascade of A431 cells. PMID:22934183

  6. Revision of Microplitis species from China with description of a new species.

    PubMed

    Zhang, Wangzhen; Song, Dongbao; Chen, Jiahua

    2017-02-09

    Microplitis bicoloratus Chen 2004 is a synonym of Microplitis prodeniae Rao & Kurian 1950, and is also the junior homonym of Microplitis bicoloratus Xu & He 2003. A new species, Microplitis fujianica sp. nov. is described and illustrated. The new species is compared with its related species from the Oriental region.

  7. Characterization of growth inhibition of oral bacteria by sophorolipid using a microplate-format assay.

    PubMed

    Solaiman, Daniel K Y; Ashby, Richard D; Uknalis, Joseph

    2017-05-01

    Sophorolipid (SL) is a class of glycolipid biosurfactant produced by yeast and has potent antimicrobial activity against many microorganisms. In this paper, a microplate-based method was developed to characterize the growth inhibition by SL on five representative species of caries-causing oral bacteria. Bacterial growth on microplate in the absence and presence of varying concentrations of SL was continuously monitored by recording the absorbance at 600nm of the cultures using a microplate reader. The results showed that SL completely inhibited the growth of the Lactobacilli at ≥1mg/ml and the Streptococci at much lower concentrations of ≥50μg/ml. More importantly, we further defined the mechanism of antimicrobial activity of SL by analyzing the pattern of the cell growth curves. SL at sublethal concentrations (<1mg/ml) is bactericidal towards the Lactobacilli; it lengthens the apparent cell-doubling time (Td) and decreases the final cell density (as indicated by A600nm) in a concentration-dependent manner. Against the oral Streptococci, on the other hand, SL at sublethal concentrations (<50μg/ml) is bacteriostatic; it delays the onset of cell growth in a concentration-dependent fashion, but once the cell growth is commenced there is no noticeable adverse effect on Td and the final A600nm. Scanning electron microscopic (SEM) study of L. acidophilus grown in sublethal concentration of SL reveals extensive structural damage to the cells. S. mutans grown in sublethal level of SL did not show morphological damage to the cells, but numerous protruding structures could be seen on the cell surface. At the respective lethal levels of SL, L. acidophilus cells were lysed (at 1mg/ml SL) and the cell surface structure of S. mutans (at 130μg/ml SL) was extensively deformed. In summary, this paper presents the first report on a detailed analysis of the effects of SL on Lactobacilli and Streptococci important to oral health and hygiene.

  8. A high-throughput approach for the determination of pesticide residues in cucumber samples using solid-phase microextraction on 96-well plate.

    PubMed

    Bagheri, Habib; Es'haghi, Ali; Es-haghi, Ali; Mesbahi, Noushin

    2012-08-31

    A high-throughput solid-phase microextraction (SPME) on 96-well plate together with gas chromatography-mass spectrometry (GC-MS) was developed for the determination of some selected pesticides in cucumber samples. Pieces with the length of 1.0 cm of silicon tubing were precisely prepared and then coated on the end part of stainless steel wires. The prepared fibers were positioned in a home-made polytetrafluoroethylene (PTFE)-based constructed ninety-six holes block to have the possibility of simultaneous immersion of the SPME fibers into the center of individual wells. Pesticides such as diazinon, penconazol, tebuconazol, bitertanol, malathion, phosalone and chlorpyrifos-methyl were selected for their highly application in cucumber field. The performances of the SPME fibers, such as intra and inter-fibers reproducibility, were evaluated and the results showed a good similarity in extraction yields. A volume of 1 mL of the aquatic supernatant of the cucumber samples was transferred into the 96-well plate and the array of SPME fibers was applied for the extraction of the selected pesticides. The important parameters influencing the whole extraction process including, organic solvent percent, salt addition, dilution factor, stirring rate and extraction time were optimized. The inter- and intra-day RSD% were found to be less than 15.4%. Limits of detection (LOD) and limits of quantification (LOQ) were below 60 and 180 μg kg(-1), respectively. The coefficient of determination was satisfactory (r(2)>0.99) for all the studied analytes. The developed method was successfully applied to the monitoring of several samples gathered from local markets.

  9. Development of cystic embryoid bodies with visceral yolk-sac-like structures from mouse embryonic stem cells using low-adherence 96-well plate.

    PubMed

    Yasuda, Emiko; Seki, Yuji; Higuchi, Takatoshi; Nakashima, Fumio; Noda, Tomozumi; Kurosawa, Hiroshi

    2009-04-01

    Cystic embryoid bodies with visceral yolk-sac-like structure (cystic EB-Vs) are used as a model for the study of early extraembryonic tissue formation containing visceral endoderm-like derivatives. In this study, we optimized the cell density of embryonic stem (ES) cells for developing cystic EB-Vs in a low-adherence 96-well plate. When ES cells were seeded at a density of 4000 cells/well, the cystic EB-Vs were most efficiently developed from ES cells via forming multicellular spherical aggregates called embryoid bodies (EBs). The suspension culture in the low-adherence plate was preferable for developing EBs into cystic EB-Vs rather than the attachment culture in the plate coated with 0.1% gelatin. The seeding cell density of 4000 cells/well was always superior to 1000 cells/well in the efficiency of cystic EB-V development. Because the high-cell density culture generally raises the limitation of oxygen and nutrient supplies, we investigated the effects of low-oxygen and low-nutrient conditions on the development of cystic EB-Vs. It was found that low oxygen tension was not a factor for promoting the development of cystic EB-Vs. It was suggested that a low-nutrient medium is preferred for developing cystic EB-Vs rather than a sufficient-nutrient medium. In conclusion, the suspension culture in the low-adherence 96-well plate seeded with 4000 ES cells/well was optimum for developing cystic EB-Vs. The low-nutrient condition may be one of the factors for promoting the development of cystic EB-Vs.

  10. High-throughput determination of fudosteine in human plasma by liquid chromatography-tandem mass spectrometry, following protein precipitation in the 96-well plate format.

    PubMed

    Wen, Jun; Wu, Yiwen; Zhang, Linli; Qi, Yunpeng; Fan, Guorong; Wu, Yutian; Li, Zhen

    2008-05-01

    A 96-well protein precipitation, liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and fully validated for the determination of fudosteine in human plasma. After protein precipitation of the plasma samples (50 microL) by the methanol (150 microL) containing the internal standard (IS), erdosteine, the 96-well plate was vortexed for 5 min and centrifuged for 15 min. The 100 microL supernatant and 100 microL mobile phase were added to another plate and mixed and then the mixture was directly injected into the LC-MS/MS system in the negative ionization mode. The separation was performed on a XB-CN column for 3.0 min per sample using an eluent of methanol-water (60:40, v/v) containing 0.005% formic acid. Multiple reaction monitoring (MRM) using the precursor-product ion transitions m/z 178-->91 and m/z 284-->91 was performed to quantify fudosteine and erdosteine, respectively. The method was sensitive with a lower limit of quantification (LLOQ) of 0.02 microg mL(-1), with good linearity (r>0.999) over the linear range of 0.02-10 microg mL(-1). The within- and between-run precision was less than 5.5% and accuracy ranged from 94.2 to 106.7% for quality control (QC) samples at three concentrations of 0.05, 1 and 8 microg mL(-1). The method was employed in the clinical pharmacokinetic study of fudosteine formulation product after oral administration to healthy volunteers.

  11. Reader Response in Perspective.

    ERIC Educational Resources Information Center

    Sloan, Glenna

    2002-01-01

    Presents an overview of critical theory to show how the method of Reader Response evolved. Discusses theories of reading and the reader; variations within reader response; and implementing reader response in literacy programs. Includes a brief response by Robert E. Probst. (RS)

  12. A new cadmium reduction device for the microplate determination of nitrate in water, soil, plant tissue, and physiological fluids.

    PubMed

    Crutchfield, James D; Grove, John H

    2011-01-01

    A reusable catalytic reductor consisting of 96 copperized-cadmium pins attached to a microplate lid was developed to simultaneously reduce nitrate (NO3-) to nitrite (NO2-) in all wells of a standard microplate. The resulting NO2- is analyzed colorimetrically by the Griess reaction using a microplate reader. Nitrate data from groundwater samples analyzed using the new device correlated well with data obtained by ion chromatography (r2 = 0.9959). Soil and plant tissue samples previously analyzed for NO3- in an interlaboratory validation study sponsored by the Soil Science Society of America were also analyzed using the new technique. For the soil sample set, the data are shown to correlate well with the other methods used (r2 = 0.9976). Plant data correlated less well, especially for samples containing low concentrations of NO3-. Reasons for these discrepancies are discussed, and new techniques to increase the accuracy of the analysis are explored. In addition, a method is presented for analyzing NO3- in physiological fluids (blood serum and urine) after matrix modification with Somogyi's reagent. A protocol for statistical validation of data when analyzing samples with complex matrixes is also established. The simplicity, adaptability, and low cost of the device indicate its potential for widespread application.

  13. Rapid quantitative analysis of lipids using a colorimetric method in a microplate format.

    PubMed

    Cheng, Yu-Shen; Zheng, Yi; VanderGheynst, Jean S

    2011-01-01

    A colorimetric sulfo-phospho-vanillin (SPV) method was developed for high throughput analysis of total lipids. The developed method uses a reaction mixture that is maintained in a 96-well microplate throughout the entire assay. The new assay provides the following advantages over other methods of lipid measurement: (1) background absorbance can be easily corrected for each well, (2) there is less risk of handling and transferring sulfuric acid contained in reaction mixtures, (3) color develops more consistently providing more accurate measurement of absorbance, and (4) the assay can be used for quantitative measurement of lipids extracted from a wide variety of sources. Unlike other spectrophotometric approaches that use fluorescent dyes, the optimal spectra and reaction conditions for the developed assay do not vary with the sample source. The developed method was used to measure lipids in extracts from four strains of microalgae. No significant difference was found in lipid determination when lipid content was measured using the new method and compared to results obtained using a macro-gravimetric method.

  14. Microplate array diagonal gel electrophoresis for cohort studies of microsatellite loci.

    PubMed

    Chen, Xiao-he; O'Dell, Sandra D; Day, Ian N M

    2002-05-01

    After PCR amplification, we have achieved precise sizing of trinucleotide and tetranucleotide microsatellite alleles on 96-well open-faced polyacrylamide microplate array diagonal gel electrophoresis (MADGE) gels: two tetranucleotide repeats, HUMTHOI (five alleles 248-263 bp) and DYS390 (eight alleles 200-228 bp), and DYS392, a trinucleotide repeat (eight alleles 210-231 bp). A gel matrix of Duracryl, a high mechanical strength polyacrylamide derivative, and appropriate ionic conditions provide the 1.3%-1.5% band resolution required. No end-labeling of primers is needed, as the sensitive Vistra Green intercalating dye is used for the visualization of bands. Co-run markers bracketing the PCR fragments ensure accurate sizing without inter-lane variability. Electrophoresis of multiple gels in a thermostatically controlled tank allows up to 1000 samples to be run in 90 min. Gel images were analyzed using a Fluorlmager 595 fluorescent scanning system, and alleles were identified using Phoretix software for band migration measurement and Microsoft Excel to compute fragment sizes. Estimated sizes were interpolated precisely to achieve accurate binning. Microsatellite-MADGE represents a utilitarian methodfor high-throughput genotyping in cohort studies, using standard laboratory equipment.

  15. Evaluation of drug load and polymer by using a 96-well plate vacuum dry system for amorphous solid dispersion drug delivery.

    PubMed

    Chiang, Po-Chang; Ran, Yingqing; Chou, Kang-Jye; Cui, Yong; Sambrone, Amy; Chan, Connie; Hart, Ryan

    2012-06-01

    It is well recognized that poor dissolution rate and solubility of drug candidates are key limiting factors for oral bioavailability. While numerous technologies have been developed to enhance solubility of the drug candidates, poor water solubility continuously remains a challenge for drug delivery. Among those technologies, amorphous solid dispersions (SD) have been successfully employed to enhance both dissolution rate and solubility of poorly water-soluble drugs. This research reports a high-throughput screening technology developed by utilizing a 96-well plate system to identify optimal drug load and polymer using a solvent casting approach. A minimal amount of drug was required to evaluate optimal drug load in three different polymers with respect to solubility improvement and solid-state stability of the amorphous drug-polymer system. Validation of this method was demonstrated with three marketed drugs as well as with one internal compound. Scale up of the internal compound SD by spray drying further confirmed the validity of this method, and its quality was comparable to a larger scale process. Here, we demonstrate that our system is highly efficient, cost-effective, and robust to evaluate the feasibility of spray drying technology to produce amorphous solid dispersions.

  16. A semi-automated 96-well protein precipitation method for the determination of montelukast in human plasma using high performance liquid chromatography/fluorescence detection.

    PubMed

    Kitchen, Chester J; Wang, Amy Q; Musson, Donald G; Yang, Amy Y; Fisher, Alison L

    2003-03-26

    A simple, semi-automated, protein precipitation assay for the determination of montelukast (SINGULAIR, MK-0476) in human plasma has been developed. Montelukast is a potent and selective antagonist of the cysteinyl leukotriene receptor used for the treatment of asthma. A Packard MultiPROBE II EX is used to transfer 300 microl of plasma from sample, standard, and QC sample tubes to a microtiter plate (96-well). After addition of the internal standard by a repeating pipettor, a Tomtec QUADRA 96 adds 400 microl of acetonitrile to all plasma sample wells, simultaneously, in the microtiter plate. The Tomtec is also used to transfer the acetonitrile supernatant from the plasma protein precipitation step, batchwise, to another microtiter plate for analysis by HPLC with fluorescence detection. This assay has been validated and implemented for a clinical study of over 1300 plasma samples and is comparable to manual assays in the LLOQ (lower limit of quantitation, 3 ng/ml) and in stability. This is the first semi-automated protein precipitation assay published for the analysis of montelukast in human plasma and it results in significant time savings over the manual methods, both in sample preparation and in HPLC run time.

  17. Induction of apoptosis by intracellular potassium ion depletion: using the fluorescent dye PBFI in a 96-well plate method in cultured lung cancer cells.

    PubMed

    Andersson, B; Janson, V; Behnam-Motlagh, P; Henriksson, R; Grankvist, K

    2006-09-01

    Depletion of intracellular potassium ions (K+) is necessary for cells to shrink, activate caspases and induce DNA fragmentation, events which are features of apoptosis. Here we describe a 96-well plate method using the cell permeable form of K+ binding benzofuran isophtalate (PBFI-AM) to measure intracellular K+ content in relation to untreated control. Cultured human pulmonary mesothelioma cells (P31) and small-cell lung cancer cells (U1690) were treated with K+ flux modulators in order to deprive the cells of intracellular K+. The combination of K+ influx inhibition with 10 micromol/L bumetanide plus 10 micromol/L ouabain and K+ efflux stimulation with 3 mg/L amphotericin B or 5 micromol/L nigericin efficiently reduced the intracellular K+ content after 3 h. Manipulation of K+ fluxes with subsequent intracellular K+ depletion induced apoptosis of lung cancer cells, as detected by caspase-3 activity after 3 h K+ depletion followed by 24 h proliferation and TUNEL positive staining after 48 h proliferation. We concluded that the PBFI-AM assay was a useful tool to determine intracellular K+ content in relation to untreated control, and that intracellular K+ depletion of lung cancer cells by clinically used drugs of relevant concentrations induced apoptosis. These findings may lead to novel therapeutic strategies in the treatment of lung cancer.

  18. Simplified method for determination of clarithromycin in human plasma using protein precipitation in a 96-well format and liquid chromatography-tandem mass spectrometry.

    PubMed

    Shin, Jaekyu; Pauly, Daniel F; Johnson, Julie A; Frye, Reginald F

    2008-08-01

    A simplified method to determine clarithromycin concentrations in human plasma using protein precipitation in a 96-well plate and liquid chromatography-tandem mass spectrometry was developed and validated. Plasma proteins were precipitated with acetonitrile and roxithromycin was used as the internal standard. After vortex mixing and centrifugation, the supernatants were directly injected onto a Phenomenex Luna Phenyl-Hexyl column (50 mm x 2.0 mm ID, 3 microm). The mobile phase consisted of water and methanol (30:70, v/v) containing 0.1% formic acid and 5mM ammonium acetate. The flow rate was 0.22 mL/min and the total run time (injection to injection) was less than 3 min. Detection of the analytes was achieved using positive ion electrospray tandem mass spectrometry in selected reaction monitoring (SRM) mode. The linear standard curve ranged from 100 to 5000 ng/mL and the precision and accuracy (inter- and intra-run) were within 7.9% and 4.9%, respectively. The method was successfully used to determine clarithromycin concentrations in human plasma samples obtained from healthy subjects who were given clarithromycin 500 mg for 3 days. The method is rapid, simple, precise and directly applicable to clarithromycin pharmacokinetic studies.

  19. Soluble penicillin-binding protein 2a: beta-lactam binding and inhibition by non-beta-lactams using a 96-well format.

    PubMed

    Toney, J H; Hammond, G G; Leiting, B; Pryor, K D; Wu, J K; Cuca, G C; Pompliano, D L

    1998-01-01

    High level methicillin resistance in Staphylococcus aureus is dependent upon the acquisition of the mecA gene encoding penicillin-binding protein 2a (PBP2a). PBP2a is a member of a family of peptidoglycan biosynthetic enzymes involved in assembly of the cell wall in bacteria and is poorly inactivated by beta-lactam antibiotics. We describe a 96-well-filter binding assay using recombinant, soluble PBP2a which allows for kinetic measurement of penicillin binding. The deacylation rate constant for the PBP2a-penicillin G covalent complex was found to be 5.7 +/- 1.0 x 10(-5) s-1 at 30 degrees C (half-life of approximately 200 min). For the PBP2a acylation reaction, the value of K(m) (penicillin G) = 0.5 +/- 0.1 mM and kcat = 1 x 10(-3) s-1, which yields a second-order rate constant (kcat/K(m)) for inactivation of 2.0 M-1 s-1. Using this assay, several non-beta-lactam inhibitors including Cibacron blue have been found which exhibit IC50 values between 10 and 30 microM. The binding affinities of several carbapenems and beta-lactams correlated well between the filter binding assay described in this report and an electrophoretic assay for PBP2a using membranes prepared form methicillin-resistant S. aureus.

  20. Method development for the quantitation of ABT-578 in rabbit artery tissue by 96-well liquid-liquid extraction and liquid chromatography/tandem mass spectrometric detection.

    PubMed

    Ji, Qin C; Zhang, Jun; Rodila, Ramona; Watson, Pamela; El-Shourbagy, Tawakol

    2004-01-01

    Quantitative determination of drug concentrations in tissue samples can provide critical information for drug metabolism, kinetics, and toxicity evaluations. For analysis of tissue samples using liquid chromatography/tandem mass spectrometric (LC/MS/MS) detection, homogenization is a critical step in achieving good assay performance. Assay performance can be closely evaluated by spiking the drug directly into tissue samples prior to homogenization. It is especially important to include this assay evaluation for the analysis of artery tissue samples because artery tissue is very elastic, making it quite a challenge to develop an effective procedure for homogenization. An LC/MS/MS assay in 96-well format using liquid-liquid extraction was developed for analyzing ABT-578 in rabbit artery samples. Tissue quality control samples were prepared by spiking ABT-578 stock solutions directly into the tissue before homogenization. The usage of the tissue control samples gives a thorough evaluation of the sample preparation process that includes both homogenization and sample extraction. A 20% blood in saline solution was used as a homogenization solution. Calibration standards were made by spiking ABT-578 into rabbit whole blood. Blood quality control samples were also prepared by spiking ABT-578 into rabbit whole blood. These blood QC samples were used to confirm the validity of the calibration curve. A lower limit of quantitation of 0.050 ng/mL was achieved. The linear dynamic range of blood standards was from 0.050-30.3 ng/mL with the correlation coefficient (r) ranging from 0.9969-0.9996. Overall %CV was between 1.3 and 7.0%, and analytical recovery was between 98.2 and 105.8% for blood QC samples. The %CVs for tissue QC samples were between 6.7 and 13.0%, and analytical recovery after correction was between 93.5 and 114.3%.

  1. A high-throughput protein refolding screen in 96-well format combined with design of experiments to optimize the refolding conditions.

    PubMed

    Dechavanne, Vincent; Barrillat, Nicolas; Borlat, Frederic; Hermant, Aurélie; Magnenat, Laurent; Paquet, Mikael; Antonsson, Bruno; Chevalet, Laurent

    2011-02-01

    Production of correctly folded and biologically active proteins in Escherichiacoli can be a challenging process. Frequently, proteins are recovered as insoluble inclusion bodies and need to be denatured and refolded into the correct structure. To address this, a refolding screening process based on a 96-well assay format supported by design of experiments (DOE) was developed for identification of optimal refolding conditions. After a first generic screen of 96 different refolding conditions the parameters that produced the best yield were further explored in a focused DOE-based screen. The refolding efficiency and the quality of the refolded protein were analyzed by RP-HPLC and SDS-PAGE. The results were analyzed by the DOE software to identify the optimal concentrations of the critical additives. The optimal refolding conditions suggested by DOE were verified in medium-scale refolding tests, which confirmed the reliability of the predictions. Finally, the refolded protein was purified and its biological activity was tested in vitro. The screen was applied for the refolding of Interleukin 17F (IL-17F), stromal-cell-derived factor-1 (SDF-1α/CXCL12), B cell-attracting chemokine 1 (BCA-1/CXCL13), granulocyte macrophage colony stimulating factor (GM-CSF) and the complement factor C5a. This procedure identified refolding conditions for all the tested proteins. For the proteins where refolding conditions were already available, the optimized conditions identified in the screening process increased the yields between 50% and 100%. Thus, the method described herein is a useful tool to determine the feasibility of refolding and to identify high-yield scalable refolding conditions optimized for each individual protein.

  2. Phospholipid removal combined with a semi-automated 96-well SPE application for determination of budesonide in human plasma with LC-MS/MS.

    PubMed

    Nilsson, Karin; Andersson, Maria; Beck, Olof

    2014-11-01

    A semi-automated method for quantification of budesonide in human plasma was developed, validated, and applied for high-volume analysis of samples in connection with a pharmacokinetic study. Protein and phospholipid removal was performed using an Ostro 96-well filter plate and subsequently combined with C18 solid-phase extraction on a Hamilton Microlab STARlet automation robot. The final extracts were evaporated to dryness and redissolved in 20% acetonitrile/water. The procedure used budesonide-d8 as internal standard and gave a 3.5-fold concentration of plasma to extract. The final extracts (5 μL injected) were analyzed with selected reaction monitoring liquid chromatography-tandem mass spectrometry (LC-MS/MS) using electrospray ionization in positive mode. The chromatography system used a 100 mm ACQUITY BEH UPLC column and a gradient system consisting of aqueous 0.1% formic acid and acetonitrile as organic modifier. Phospholipid removal was found to be needed during method development in order to reduce ion suppression effects from matrix and to increase method sensitivity. The measuring range was 50-5000 pg/mL with and LOD 24 pg/mL. Calibration response showed good linearity (correlation coefficients<0.99) over the measuring range. The absolute recovery over the sample preparation procedure was estimated to 67%. Total imprecision was <9% at three levels and accuracy was between 98.9 and 103%. The method was successfully applied for analysis of 864 study samples in a short time. The quality control samples at concentration levels 200 and 2000 pg/mL gave a total imprecision of 7.4% and 4.2%, respectively, (n=95).

  3. Absorbance and fluorometric sensing with capillary wells microplates

    SciTech Connect

    Tan, Han Yen; Cheong, Brandon Huey-Ping; Neild, Adrian; Wah Ng, Tuck; Liew, Oi Wah

    2010-12-15

    Detection and readout from small volume assays in microplates are a challenge. The capillary wells microplate approach [Ng et al., Appl. Phys. Lett. 93, 174105 (2008)] offers strong advantages in small liquid volume management. An adapted design is described and shown here to be able to detect, in a nonimaging manner, fluorescence and absorbance assays minus the error often associated with meniscus forming at the air-liquid interface. The presence of bubbles in liquid samples residing in microplate wells can cause inaccuracies. Pipetting errors, if not adequately managed, can result in misleading data and wrong interpretations of assay results; particularly in the context of high throughput screening. We show that the adapted design is also able to detect for bubbles and pipetting errors during actual assay runs to ensure accuracy in screening.

  4. JAPANESE READERS (TITLE SUPPLIED).

    ERIC Educational Resources Information Center

    THREE JAPANESE READERS ARE PRESENTED WHICH CONTAIN VOCABULARY, NOTES, AND DRILL SENTENCES. THE THREE READERS--(1) ARU SARARIIMAN NO ITI-NITI, (2) OTOOSAN WA KAMI-SAMA, AND (3) ARU GAKUSEI NO HANNITI--ARE WRITTEN IN THE ROMAJI ALPHABET. EACH READER HAS A VOCABULARY LISTING WITH ENGLISH EXPLANATIONS AND DRILL SENTENCES. AN ENGLISH WORD DEFINITION…

  5. Responding to Readers.

    ERIC Educational Resources Information Center

    Gill, Sharon Ruth

    2002-01-01

    Provides classroom teachers with seven guidelines for responding to readers in ways that support the use of strategies for making sense of text. Discusses traditional responses to readers and "round-robin" reading versus reading conferences. Concludes that by responding appropriately to readers, teachers provide powerful demonstrations of the…

  6. A 96-well single-pot liquid-liquid extraction, hydrophilic interaction liquid chromatography-mass spectrometry method for the determination of muraglitazar in human plasma.

    PubMed

    Xue, Y-J; Liu, Jane; Unger, Steve

    2006-06-07

    A single-pot liquid-liquid extraction (LLE) with hydrophilic interaction liquid chromatography/tandem mass spectrometry (HILIC/MS/MS) method has been developed and validated for the determination of muraglitazar, a hydrophobic diabetes drug, in human plasma. To 0.050 ml of each plasma sample in a 96-well plate, the internal standard solution in acetonitrile and toluene were added to extract the compound of interest. The plate was vortexed, followed by centrifugation. The organic layer was then directly injected into an LC/MS/MS system. Chromatographic separation was achieved isocratically on a Thermohypersil_Keystone, Hypersil silica column (3 mmx50 mm, 3 microm). The mobile phase contained 85% of methyl t-butyl ether and 15% of 90/10 (v/v) acetonitrile/water with 0.3% trifluoroacetic acid. Post-column mobile phase of 50/50 (v/v) acetonitrile/water containing 0.1% formic acid was added. Detection was by positive ion electrospray tandem mass spectrometry on a Sciex API 4000. The standard curve, ranged from 1 to 1000 ng/ml, was fitted to a 1/x weighted quadratic regression model. This single-pot LLE approach effectively eliminated time-consuming organic layer transfer, dry-down, and sample reconstitution steps, which are essential for a conventional liquid-liquid extraction procedure. The modified mobile phase was more compatible with the direct injection of the commonly used extraction solvents in LLE. Furthermore, the modified mobile phase improved the retention of muraglitazar, a hydrophobic compound, on the normal phase silica column. The validation results demonstrated that this method was rugged and suitable for analyzing muraglitazar in human plasma. In comparison with a revised-phase LC/MS/MS method, this single-pot LLE, HILIC/MS/MS method improved the detection sensitivity by more than four-fold based upon the LLOQ signal to noise ratio. This approach may be applied to other hydrophobic compounds with proper modification of the mobile phase compositions.

  7. Microplates in liquid chromatography--new solution in clinical research? A review.

    PubMed

    Krcmova, Lenka; Solichova, Dagmar; Solich, Petr

    2013-10-15

    Microplates are routinely used in Radio- or Immuno-assays. Recently, microplates have found use not only in analytical but also in the pre-analytical phase in bioanalyses (sample storage, sample preparation). New connection of this technology to liquid chromatography could be economical, fast and simple solution for many routine laboratories handling large sequences of biological samples. This review summarises the application of microplates in bioanalytical laboratories. Different types of sorbents, materials and shapes of microplates are discussed, and the main advantages and disadvantages of microplates used in clinical research are presented.

  8. A Sensitive Microplate Assay for Lipase Activity Measurement Using Olive Oil Emulsion Substrate: Modification of the Copper Soap Colorimetric Method.

    PubMed

    Mustafa, Ahmad; Karmali, Amin; Abdelmoez, Wael

    2016-01-01

    The present work involves a sensitive high-throughput microtiter plate based colorimetric assay for estimating lipase activity using cupric acetate pyridine reagent (CAPR). In the first approach, three factors two levels factorial design methodology was used to evaluate the interactive effect of different parameters on the sensitivity of the assay method. The optimization study revealed that the optimum CAPR concentration was 7.5% w/v, the optimum solvent was heptane and the optimum CAPR pH was 6. In the second approach, the optimized colorimetric microplate assay was used to measure lipase activity based on enzymatic hydrolysis of olive oil emulsion substrate at 37°C and 150 rpm. The emulsion substrates were formulated by using olive oil, triton X-100 (10% v/v in pH 8) and sodium phosphate buffer of pH 8 in ratio of 1:1:1 in the case of Candida sp. lipase. While in the case of immobilized lipozyme RMIM, The emulsion substrates were formulated by using olive oil, triton X-100 (1% v/v in pH 8) and sodium phosphate buffer of pH 8 in ratio of 2:1:1. Absorbance was measured at 655 nm. The stability of this assay (in terms of colored heptane phase absorbance readings) retained more than 92.5% after 24 h at 4°C compared to the absorbance readings measured at zero time. In comparison with other lipase assay methods, beside the developed sensitivity, the reproducibility and the lower limit of detection (LOD) of the proposed method, it permits analyzing of 96 samples at one time in a 96-well microplate. Furthermore, it consumes small quantities of chemicals and unit operations.

  9. Enhanced dual-frequency operation of a polymerized liquid crystal microplate by liquid crystal infiltration

    NASA Astrophysics Data System (ADS)

    Kumagai, Takayuki; Yoshida, Hiroyuki; Ozaki, Masanori

    2017-04-01

    The electric-field-induced switching behavior of a polymer microplate is investigated. A microplate fabricated with a photopolymerizable dual-frequency liquid crystal was surrounded by an unpolymerized photopolymerizable dual-frequency liquid crystal in the isotropic phase. As an electric field was applied along the plane of the microplate, the microplate switched to set its interior molecular orientation to be either parallel or perpendicular to the field, depending on the frequency. Analysis of the rotational behavior, as well as numerical calculations, showed that the surrounding unpolymerized photopolymerizable dual-frequency liquid crystal infiltrated into the microplate, which enhanced the dielectric properties of the microplate. To the best of our knowledge, this is the first report of an enhanced dual-frequency dielectric response of a polymer microplate induced by liquid crystal infiltration.

  10. Automated panning and screening procedure on microplates for antibody generation from phage display libraries.

    PubMed

    Turunen, Laura; Takkinen, Kristiina; Söderlund, Hans; Pulli, Timo

    2009-03-01

    Antibody phage display technology is well established and widely used for selecting specific antibodies against desired targets. Using conventional manual methods, it is laborious to perform multiple selections with different antigens simultaneously. Furthermore, manual screening of the positive clones requires much effort. The authors describe optimized and automated procedures of these processes using a magnetic bead processor for the selection and a robotic station for the screening step. Both steps are performed in a 96-well microplate format. In addition, adopting the antibody phage display technology to automated platform polyethylene glycol precipitation of the enriched phage pool was unnecessary. For screening, an enzyme-linked immunosorbent assay protocol suitable for a robotic station was developed. This system was set up using human gamma-globulin as a model antigen to select antibodies from a VTT naive human single-chain antibody (scFv) library. In total, 161 gamma-globulin-selected clones were screened, and according to fingerprinting analysis, 9 of the 13 analyzed clones were different. The system was further tested using testosterone bovine serum albumin (BSA) and beta-estradiol-BSA as antigens with the same library. In total, 1536 clones were screened from 4 rounds of selection with both antigens, and 29 different testosterone-BSA and 23 beta-estradiol-BSA binding clones were found and verified by sequencing. This automated antibody phage display procedure increases the throughput of generating wide panels of target-binding antibody candidates and allows the selection and screening of antibodies against several different targets in parallel with high efficiency.

  11. Teachers as Readers: Building Communities of Readers

    ERIC Educational Resources Information Center

    Cremin, Teresa; Mottram, Marilyn; Collins, Fiona; Powell, Sacha; Safford, Kimberly

    2009-01-01

    Given the narrow scope of primary teachers' knowledge and use of children's literature identified in Phase I of "Teachers as Readers" (2006-2007), the core goal of the Phase II project was to improve teachers' knowledge and experience of such literature in order to help them increase children's motivation and enthusiasm for reading,…

  12. Microform Reader Maintenance.

    ERIC Educational Resources Information Center

    Hall, Hal W.; Michaels, George H.

    1985-01-01

    Describes experiences in organizing a program of microform reader and reader/printer maintenance at Texas A & M's Sterling C. Evans Library and offers guidelines for regular machine maintenance and repair. Guidelines discussed relate to maintenance philosophy, general machine cleaning, troubleshooting, service contracts, supplies,…

  13. The Readers' Advisor's Companion.

    ERIC Educational Resources Information Center

    Shearer, Kenneth D., Ed.; Burgin, Robert, Ed.

    From scholarly discourse to the latest issues in readers' advisory services, this guide provides up-to-date information on the many challenges of the practice, as well as on interdisciplinary directions, continuing education, and the gap in graduate professional education for readers' advisory. The book's 16 chapters are organized into three main…

  14. Romancing the YA Reader.

    ERIC Educational Resources Information Center

    Charles, John; Mosley, Shelley; Bouricius, Ann

    1999-01-01

    Discusses the romance genre and provides guidelines for librarians with high school aged readers. Highlights include where to find information about the romance genre; the appeal of romance fiction; levels of sex and sensuality; reference tools; subgenres; and authors and titles that may be popular with young adult romance readers. (LRW)

  15. Texts and Readers.

    ERIC Educational Resources Information Center

    Iser, Wolfgang

    1980-01-01

    Notes that, since fictional discourse need not reflect prevailing systems of meaning and norms or values, readers gain detachment from their own presuppositions; by constituting and formulating text-sense, readers are constituting and formulating their own cognition and becoming aware of the operations for doing so. (FL)

  16. Becoming a Reader.

    ERIC Educational Resources Information Center

    Causey, John P.; Pfau, Donald W.

    Reflecting an orientation to reading, language development, and students, this guide was drawn together to help give focus and direction to the efforts that are occurring in schools where people are studying the environment in which a child becomes a reader. An overview of the guide centers around the belief that a child will become a reader if he…

  17. Developing Strategic Readers

    ERIC Educational Resources Information Center

    Jones, Jennifer; Leahy, Susie

    2006-01-01

    As elementary students progress from "learning to read" toward "reading to learn," it is vital that they become strategic readers. A strategic reader is one who understands when and how to use a strategy in order to comprehend text. While some students use strategies naturally during the reading process, most students must be taught how to…

  18. Science Fiction Readers Theatre.

    ERIC Educational Resources Information Center

    Fredericks, Anthony D.

    This book presents science fiction readers theatre scripts for use with grades 4 through 8. The introduction provides information on what readers theatre is and its value, as well as presentation suggestions (e.g., preparing scripts, staging, props, delivery, and post-presentation). Each script includes staging directions and ideas for learning…

  19. Development of a cloud point extraction and spectrophotometry-based microplate method for the determination of nitrite in human urine and blood.

    PubMed

    Zhao, Jiao; Lu, Yunhui; Fan, Chongyang; Wang, Jun; Yang, Yaling

    2015-02-05

    A novel and simple method for the sensitive determination of trace amounts of nitrite in human urine and blood has been developed by combination of cloud point extraction (CPE) and microplate assay. The method is based on the Griess reaction and the reaction product is extracted into nonionic surfactant Triton-X114 using CPE technique. In this study, decolorization treatment of urine and blood was applied to overcome the interference of matrix and enhance the sensitivity of nitrite detection. Multi-sample can be simultaneously detected thanks to a 96-well microplate technique. The effects of different operating parameters such as type of decolorizing agent, concentration of surfactant (Triton X-114), addition of (NH4)2SO4, extraction temperature and time, interfering elements were studied and optimum conditions were obtained. Under the optimum conditions, a linear calibration graph was obtained in the range of 10-400 ng mL(-1) of nitrite with limit of detection (LOD) of 2.5 ng mL(-1). The relative standard deviation (RSD) for determination of 100 ng mL(-1) of nitrite was 2.80%. The proposed method was successfully applied for the determination of nitrite in the urine and blood samples with recoveries of 92.6-101.2%.

  20. Development of a cloud point extraction and spectrophotometry-based microplate method for the determination of nitrite in human urine and blood

    NASA Astrophysics Data System (ADS)

    Zhao, Jiao; Lu, Yunhui; Fan, Chongyang; Wang, Jun; Yang, Yaling

    2015-02-01

    A novel and simple method for the sensitive determination of trace amounts of nitrite in human urine and blood has been developed by combination of cloud point extraction (CPE) and microplate assay. The method is based on the Griess reaction and the reaction product is extracted into nonionic surfactant Triton-X114 using CPE technique. In this study, decolorization treatment of urine and blood was applied to overcome the interference of matrix and enhance the sensitivity of nitrite detection. Multi-sample can be simultaneously detected thanks to a 96-well microplate technique. The effects of different operating parameters such as type of decolorizing agent, concentration of surfactant (Triton X-114), addition of (NH4)2SO4, extraction temperature and time, interfering elements were studied and optimum conditions were obtained. Under the optimum conditions, a linear calibration graph was obtained in the range of 10-400 ng mL-1 of nitrite with limit of detection (LOD) of 2.5 ng mL-1. The relative standard deviation (RSD) for determination of 100 ng mL-1 of nitrite was 2.80%. The proposed method was successfully applied for the determination of nitrite in the urine and blood samples with recoveries of 92.6-101.2%.

  1. Microplate based biosensing with a computer screen aided technique.

    PubMed

    Filippini, Daniel; Andersson, Tony P M; Svensson, Samuel P S; Lundström, Ingemar

    2003-10-30

    Melanophores, dark pigment cells from the frog Xenopus laevis, have the ability to change light absorbance upon stimulation by different biological agents. Hormone exposure (e.g. melatonin or alpha-melanocyte stimulating hormone) has been used here as a reversible stimulus to test a new compact microplate reading platform. As an application, the detection of the asthma drug formoterol in blood plasma samples is demonstrated. The present system utilizes a computer screen as a (programmable) large area light source, and a standard web camera as recording media enabling even kinetic microplate reading with a versatile and broadly available platform, which suffices to evaluate numerous bioassays. Especially in the context of point of care testing or self testing applications these possibilities become advantageous compared with highly dedicated comparatively expensive commercial systems.

  2. Surface tension drawing of liquid from microplate capillary wells.

    PubMed

    Schwalb, Willem; Ng, Tuck Wah; Lye, Jonathan Kok Keung; Liew, Oi Wah; Cheong, Brandon Huey-Ping

    2012-01-01

    Pressure differentials are routinely used to actuate flow in capillaries. We advance here an alternative means of flow generation that capitalizes on the extension of a liquid bridge achieved by the drawing of a rod through the action of surface tension. This meets the exigencies of creating controllable flow using simpler and more compact means. We found the ability to generate controllable flow to be strongly affected by the liquid bridge sustaining features, and that the use of rod diameters larger than the capillary was more conducive. The extensional flow resulting from the rupture of the liquid bridge was also found to have a strong circulation component which facilitated mixing. The approach here is highly amenable for use in capillary well microplates which have significant advantages over standard microplates. The features of this approach offer usage possibilities in biochemical applications in the field, such as in the leukocyte cell adhesion and hemagglutination tests of blood samples.

  3. Cryoalgotox: Use of cryopreserved alga in a semistatic microplate test

    SciTech Connect

    Benhra, A.; Radetski, C.M.; Ferard, J.F.

    1997-03-01

    Use of cryopreserved alga Selenastrum capricornutum has been evaluated as a simple and cost-efficient procedure in a new semistatic algal ecotoxicity test. Experiments have been conducted to compare performance criteria of this method, named Cryoalgotox, versus the classic microplate test using fresh algae. Cryoalgotox 72-h 50% effective concentrations (EC50s) determined with Cd{sup 2+}, Cu{sup 2+}, Cr{sup 6+}, and atrazine were more sensitive, repeatable (low coefficients of variation), and reproducible (low time effect) than the results obtained with the classical microplate tests. The effect of storage time at {minus}80 C on the sensitivity of the algae was assessed using cadmium as a toxic reference; it was shown that algae stored at {minus}80 C over a 3-month period gave comparable toxicity results to those found with fresh algae.

  4. Hypochlorite scavenging activity of hydroxycinnamic acids evaluated by a rapid microplate method based on the measurement of chloramines.

    PubMed

    Firuzi, Omidreza; Giansanti, Luisa; Vento, Roberta; Seibert, Cathrin; Petrucci, Rita; Marrosu, Giancarlo; Agostino, Roberta; Saso, Luciano

    2003-07-01

    Scavengers of hypochlorite (XOCl) could have beneficial effects in diseases in which this oxidant plays a pathogenic role. It has been reported that ferulic acid and chlorogenic acid, the quinic ester of caffeic acid, are good hypochlorite scavengers, but a systematic evaluation of the naturally occurring hydroxycinnamic acids (HCAs), which these substances belong to, has not been performed yet. Thus, in this work we studied, by two different in-vitro methods, the antioxidant activity of five HCAs: p-coumaric acid, ferulic acid, sinapinic acid, caffeic acid and chlorogenic acid. The methods applied in this study were based on the oxidation of human serum albumin (HSA) by XOCl, a new microplate method based on the measurement of chloramines and a previously described carbonyl assay. Firstly, lysine-derived chloramines, in the presence or absence of the HCAs, were detected using 5-thio-2-nitrobenzoic acid (TNB), measuring the absorbance at 415 nm by a microplate reader. To remove excess XOCl, Trolox, a known XOCl scavenger, was added before TNB. Secondly, lysine-derived carbonyls, in the presence or absence of the HCAs, were detected by using 2,4-dinitrophenylhydrazine. Hydroxycinnamic acids appeared active (caffeic >/= sinapinic > chlorogenic congruent with ferulic > p-coumaric acid) by both methods, suggesting possible pharmacological applications for these compounds, which are present at high concentrations in the plant kingdom.

  5. Rapid determination of trace copper in animal feed based on micro-plate colorimetric reaction and statistical partitioning correction.

    PubMed

    Niu, Yiming; Wang, Jiayi; Zhang, Chi; Chen, Yiqiang

    2017-04-15

    The objective of this study was to develop a micro-plate based colorimetric assay for rapid and high-throughput detection of copper in animal feed. Copper ion in animal feed was extracted by trichloroacetic acid solution and reduced to cuprous ion by hydroxylamine. The cuprous ion can chelate with 2,2'-bicinchoninic acid to form a Cu-BCA complex which was detected with high sensitivity by micro-plate reader at 354nm. The whole assay procedure can be completed within 20min. To eliminate matrix interference, a statistical partitioning correction approach was proposed, which makes the detection of copper in complex samples possible. The limit of detection was 0.035μg/mL and the detection range was 0.1-10μg/mL of copper in buffer solution. Actual sample analysis indicated that this colorimetric assay produced results consistent with atomic absorption spectrometry analysis. These results demonstrated that the developed assay can be used for rapid determination of copper in animal feed.

  6. A Colorimetric Microplate Assay for DNA-Binding Activity of His-Tagged MutS Protein.

    PubMed

    Banasik, Michał; Sachadyn, Paweł

    2016-09-01

    A simple microplate method was designed for rapid testing DNA-binding activity of proteins. The principle of the assay involves binding of tested DNA by his-tagged protein immobilized on a nickel-coated ELISA plate, following colorimetric detection of biotinylated DNA with avidin conjugated to horseradish peroxidase. The method was used to compare DNA mismatch binding activities of MutS proteins from three bacterial species. The assay required relatively low amounts of tested protein (approximately 0.5-10 pmol) and DNA (0.1-10 pmol) and a relatively short time of analysis (up to 60 min). The method is very simple to apply and convenient to test different buffer conditions of DNA-protein binding. Sensitive colorimetric detection enables naked eye observations and quantitation with an ELISA reader. The performance of the assay, which we believe is a distinguishing trait of the method, is based on two strong and specific molecular interactions: binding of a his-tagged protein to a nickel-coated microplate and binding of biotinylated DNA to avidin. In the reported experiments, the solution was used to optimize the conditions for DNA mismatch binding by MutS protein; however, the approach could be implemented to test nucleic acids interactions with any protein of interest.

  7. An optical microplate biosensor for the detection of methyl parathion pesticide using a biohybrid of Sphingomonas sp. cells-silica nanoparticles.

    PubMed

    Mishra, Archana; Kumar, Jitendra; Melo, Jose Savio

    2017-01-15

    The previously developed Sphingomonas sp. based optical microplate biosensor for methyl parathion (MP) was good as it detected multiple samples but had poor stability and low sensitivity. The present study aims to overcome these limitations. Silica nanoparticles (Si NP) were thus functionalized with polyethyleneimine (PEI) and the functionalized silica nanoparticles ((f)Si NP) were then integrated with Sphingomonas sp. cells. The process was optimized for hydrolysis of MP into p-nitrophenol (PNP). Integration of (f)Si NP with cells was confirmed by FT-IR analysis. Biohybrid of Sphingomonas sp.-(f)Si NP was immobilized on the wells of microplate and associated directly with the optical transducer of microplate reader. Immobilized biohybrid of Sphingomonas sp.-(f)Si NP was characterized using SEM. A detection range of 0.1-1ppm MP was achieved from the linear range of calibration plot. After integration with (f)Si NP the storage stability of biohybrid was enhanced ten times from 18 to 180 days. This study proves that after interaction of cells with (f)Si NP, improved the sensitivity and stability of the biosensor. Spiked samples were also analyzed and correlated using this biohybrid based biosensor.

  8. GPS estimates of microplate motions, northern Caribbean: evidence for a Hispaniola microplate and implications for earthquake hazard

    NASA Astrophysics Data System (ADS)

    Benford, B.; DeMets, C.; Calais, E.

    2012-09-01

    We use elastic block modelling of 126 GPS site velocities from Jamaica, Hispaniola, Puerto Rico and other islands in the northern Caribbean to test for the existence of a Hispaniola microplate and estimate angular velocities for the Gônave, Hispaniola, Puerto Rico-Virgin Islands and two smaller microplates relative to each other and the Caribbean and North America plates. A model in which the Gônave microplate spans the whole plate boundary between the Cayman spreading centre and Mona Passage west of Puerto Rico is rejected at a high confidence level. The data instead require an independently moving Hispaniola microplate between the Mona Passage and a likely diffuse boundary within or offshore from western Hispaniola. Our updated angular velocities predict 6.8 ± 1.0 mm yr-1 of left-lateral slip along the seismically hazardous Enriquillo-Plantain Garden fault zone of southwest Hispaniola, 9.8 ± 2.0 mm yr-1 of slip along the Septentrional fault of northern Hispaniola and ˜14-15 mm yr-1 of left-lateral slip along the Oriente fault south of Cuba. They also predict 5.7 ± 1 mm yr-1 of fault-normal motion in the vicinity of the Enriquillo-Plantain Garden fault zone, faster than previously estimated and possibly accommodated by folds and faults in the Enriquillo-Plantain Garden fault zone borderlands. Our new and a previous estimate of Gônave-Caribbean plate motion suggest that enough elastic strain accumulates to generate one to two Mw˜ 7 earthquakes per century along the Enriquillo-Plantain Garden and nearby faults of southwest Hispaniola. That the 2010 M= 7.0 Haiti earthquake ended a 240-yr-long period of seismic quiescence in this region raises concerns that it could mark the onset of a new earthquake sequence that will relieve elastic strain that has accumulated since the late 18th century.

  9. Plasma-Treated Microplates with Enhanced Protein Recoveries and Minimized Extractables

    PubMed Central

    Weikart, Christopher M.; Klibanov, Alexander M.; Breeland, Adam P.; Taha, Ahmad H.; Maurer, Brian R.; Martin, Steven P.

    2016-01-01

    SiO2 Medical Products, Inc. (SiO) has developed a proprietary technology that greatly enhances protein recoveries and reduces extractables from commercial microplates used for bioanalytical assays and storage of biologics. SiO technology is based on plasma treatment that chemically modifies the surface of polypropylene with predominantly hydrogen-bond-acceptor uncharged polar groups. The resultant surface resists nonspecific protein adsorption over a wide range of protein concentrations, thereby eliminating the need to passivate (and hence potentially contaminate) the microplates with blocking proteins. High shelf-life stability and cleanliness of the plasma-treated microplates have been demonstrated using five different proteins for two common microplate formats. The protein recovery performance of plasma-treated microplates is found to be higher compared with commercial low-protein-binding microplates. PMID:27651466

  10. Microplates with integrated oxygen sensors for kinetic cell respiration measurement and cytotoxicity testing in primary and secondary cell lines.

    PubMed

    Deshpande, Rahul Ravi; Koch-Kirsch, Yvonne; Maas, Ruth; John, Gernot T; Krause, Christian; Heinzle, Elmar

    2005-06-01

    This paper presents a cytotoxicity and cell respiration assay that is nondestructive and kinetic. It makes use of 96-well microplates integrated with oxygen sensors. The oxygen signal monitored on-line gives an indication of the cell viability. We show its application for suspension cell lines (Chinese hamster ovary and HL60 cells) as well as adherent (Caco2 cells) and primary (rat hepatocytes) cells using well-known cytotoxic compounds (sodium azide, diclofenac, clozapine, sodium dodecyl sulfate, 2-thiouracil, tamoxifen, and tranylcypromine). The 50% lethality concentration (LC50) obtained from the assay is compared with the standard 3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyl-2H-tetrazolium bromide end-point assay. The cells can be grown directly in the plates, and the assay requires no further reagents or processing. The cells can be harvested for further analysis, if required. The on-line dynamic measurement allows the calculation of LC50 as a function of exposure time. LC50 was shown to decrease with time in HL60 cells. The dynamics of this process was considerably different for the three compounds sodium dodecyl sulfate, tamoxifen, and diclofenac, indicating a large potential of application of this method for cell death studies. The assay system can be applied to almost any cell-based systems with little adaptation. The assay is robust, flexible, and applicable for medium- to high-throughput systems requiring only minimal handling and no additional agent.

  11. Quantitative analysis of simvastatin and its beta-hydroxy acid in human plasma using automated liquid-liquid extraction based on 96-well plate format and liquid chromatography-tandem mass spectrometry.

    PubMed

    Zhang, Nanyan; Yang, Amy; Rogers, John Douglas; Zhao, Jamie J

    2004-01-27

    An assay based on automated liquid-liquid extraction (LLE) and liquid chromatography-tandem mass spectrometry (LC/MS/MS) has been developed and validated for the quantitative analysis of simvastatin (SV) and its beta-hydroxy acid (SVA) in human plasma. A Packard MultiProbe II workstation was used to convert human plasma samples collected following administration of simvastatin and quality control (QC) samples from individual tubes into 96-well plate format. The workstation was also used to prepare calibration standards and spike internal standards. A Tomtec Quadra 96-channel liquid handling workstation was used to perform LLE based on 96-well plates including adding solvents, separating organic from aqueous layer and reconstitution. SV and SVA were separated through a Kromasil C18 column (50 mm x 2 mm i.d., 5 microm) and detected by tandem mass spectrometry with a TurboIonspray interface. Stable isotope-labeled SV and SVA, 13CD(3)-SV and 13 CD(3)-SVA, were used as the internal standards for SV and SVA, respectively. The automated procedures reduced the overall analytical time (96 samples) to 1/3 of that of manual LLE. Most importantly, an analyst spent only a fraction of time on the 96-well LLE. A limit of quantitation of 50 pg/ml was achieved for both SV and SVA. The interconversion between SV and SVA during the 96-well LLE was found to be negligible. The assay showed very good reproducibility, with intra- and inter-assay precision (%R.S.D.) of less than 7.5%, and accuracy of 98.7-102.3% of nominal values for both analytes. By using this method, sample throughput should be enhanced at least three-fold compared to that of the manual procedure.

  12. Reader Response Approaches and the Growth of Readers.

    ERIC Educational Resources Information Center

    Spiegel, Dixie Lee

    1998-01-01

    Outlines what reader response might look like in the classroom. Presents four basic assumptions of reader response theory. Discusses ways reader response approaches to literature help students grow both in the depth and breadth of their responses to literature, and as strategic readers. (SR)

  13. A Cenozoic tectonic model for Southeast Asia - microplates and basins

    SciTech Connect

    Maher, K.A.

    1995-04-01

    A computer-assisted Cenozoic tectonic model was built for Southeast Asia and used to construct 23 base maps, 2 to 6 million years apart. This close temporal spacing was necessary to constrain all the local geometric shifts in a consistent and geologically feasible fashion. More than a hundred individual blocks were required to adequately treat Cenozoic microplate processes at a basic level. The reconstructions show tectonic evolution to be characterized by long periods of gradual evolution, interrupted by brief, widespread episodes of reorganization in fundamental plate geometries and kinematics. These episodes are triggered by major collisions, or by accumulation of smaller changes. The model takes into account difficulties inherent in the region. The Pacific and Indo-Australian plates and their predecessors have driven westward and northward since the late Paleozoic, towards each other and the relatively stationary backstop of Asia. Southeast Asia is therefore the result of a long-lived, complex process of convergent tectonics, making it difficult to reconstruct tectonic evolution as much of the continental margin and sea floor spreading record was erased. In addition, the region has been dominated by small-scale microplate processes with short time scales and internal deformation, taking place in rapidly evolving and more ductile buffer zones between the major rigid plate systems. These plate interaction zones have taken up much of the relative motion between the major plates. Relatively ephemeral crustal blocks appear and die within the buffer zones, or accrete to and disperse from the margins of the major plate systems. However, such microplate evolution is the dominant factor in Cenozoic basin evolution. This detailed testonic model aids in comprehension and prediction of basin development, regional hydrocarbon habitat, and petroleum systems.

  14. Validation of a fluorescence polarization assay (FPA) performed in microplates and comparison with other tests used for diagnosing B. melitensis infection in sheep and goats.

    PubMed

    Minas, A; Stournara, A; Minas, M; Stack, J; Petridou, E; Christodoulopoulos, G; Krikelis, V

    2007-03-30

    Fluorescence polarization assay (FPA) is a relatively new test for the serological diagnosis of Brucella spp. infection in animals. FPA, carried out in 96-well microplate format, was validated here for diagnosing B. melitensis infection in sheep and goats. This study included sera from 1933 sheep and goats, from animals reared in naturally infected flocks (verified by culture) and showing a positive reaction to two different tests conducted in parallel. In addition, 2154 sera originating from healthy sheep and goats, reared in areas where B. melitensis had never been isolated, were assayed. The optimum cut-off value offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp) was determined at 15 mP over the mean value of the buffer control used in each microplate as determined by receiver operating characteristic analysis. The DSn and DSp of the FPA for small ruminants carried out in microplates at this cut-off value were calculated to be 95.9% and 97.9% with 95% confidence intervals (95% CI) of 94.9-97.7% and 97.2-98.4%, respectively. The accuracy of the FPA, as expressed by determination of the area under the curve, was 0.991. Indirect ELISA and FPA tests offered the highest DSn when compared with the Rose Bengal test, the complement fixation test, the modified Rose Bengal test and competitive ELISA. The parallel or serial combination of FPA with indirect ELISA offers the highest DSn and DSp. As temperature can affect the results of the FPA, all reagents must be at the same temperature and the standard for comparison must always be read under the same conditions as the sera under test. FPA performed in microplates is a promising assay; the DSn and accuracy are better than those of the tests currently approved for diagnosing B. melitensis in small ruminants. Because of its simplicity, speed, and accuracy, this test can improve capacity for laboratory testing and the efficacy of an eradication program based on a test-and-slaughter policy.

  15. Capacitive label reader

    DOEpatents

    Arlowe, H.D.

    1983-07-15

    A capacitive label reader includes an outer ring transmitting portion, an inner ring transmitting portion, and a plurality of insulated receiving portions. A label is the mirror-image of the reader except that identifying portions corresponding to the receiving portions are insulated from only one of two coupling elements. Positive and negative pulses applied, respectively, to the two transmitting rings biased a CMOS shift register positively to either a 1 or 0 condition. The output of the CMOS may be read as an indication of the label.

  16. Capacitive label reader

    DOEpatents

    Arlowe, H.D.

    1985-11-12

    A capacitive label reader includes an outer ring transmitting portion, an inner ring transmitting portion, and a plurality of insulated receiving portions. A label is the mirror-image of the reader except that identifying portions corresponding to the receiving portions are insulated from only one of two coupling elements. Positive and negative pulses applied, respectively, to the two transmitting rings biased a CMOS shift register positively to either a 1 or 0 condition. The output of the CMOS may be read as an indication of the label. 5 figs.

  17. Capacitive label reader

    DOEpatents

    Arlowe, H. Duane

    1985-01-01

    A capacitive label reader includes an outer ring transmitting portion, an inner ring transmitting portion, and a plurality of insulated receiving portions. A label is the mirror-image of the reader except that identifying portions corresponding to the receiving portions are insulated from only one of two coupling elements. Positive and negative pulses applied, respectively, to the two transmitting rings biased a CMOS shift register positively to either a 1 or 0 condition. The output of the CMOS may be read as an indication of the label.

  18. Structural patterns and tectonic history of the Bauer microplate, Eastern Tropical Pacific

    USGS Publications Warehouse

    Eakins, B.W.; Lonsdale, P.F.

    2003-01-01

    The Bauer microplate was an independent slab of oceanic lithosphere that from 17 Ma to 6 Ma grew from 1.4 ?? 105 km2 to 1.2 ?? 106 km2 between the rapidly diverging Pacific and Nazca plates. Growth was by accretion at the lengthening and overlapping axes of the (Bauer-Nazca) Galapagos Rise (GR) and the (Pacific-Bauer) East Pacific Rise (EPR). EPR and GR axial propagation to create and rapidly grow the counter-clockwise spinning microplate occurred in two phases: (1) 17-15Ma, when the EPR axis propagated north and the GR axis propagated south around a narrow (100- to 200-km-wide) core of older lithosphere; and (2) 8-6 Ma, when rapid northward propagation of the EPR axis resumed, overlapping ???400 km of the fast-spreading Pacific-Nazca rise-crest and appending a large (200- to 400-km-wide) area of the west flank of that rise as a 'northern annex' to the microplate. Between 15 and 8 Ma the microplate grew principally by crustal accretion at the crest of its rises. The microplate was captured by the Nazca plate and the Galapagos Rise axis became extinct soon after 6 Ma, when the south end of the Pacific-Bauer EPR axis became aligned with the southern Pacific-Nazca EPR axis and its north end was linked by the Quebrada Transform to the northern Pacific-Nazca EPR axis. Incomplete multibeam bathymetry of the microplate margins, and of both flanks of the Pacific-Bauer and Bauer-Nazca Rises, together with archival magnetic and satellite altimetry data, clarifies the growth and (counter-clockwise) rotation of the microplate, and tests tectonic models derived from studies of the still active, much smaller, Easter and Juan Fernandez microplates. Our interpretations differ from model predictions in that Euler poles were not located on the microplate boundary, propagation in the 15-8 Ma phase of growth was not toward these poles, and microplate rotation rates were small (5??/m.y.) for much of its history, when long, bounding transform faults reduced coupling to Nazca plate

  19. Microplate-based method for high-throughput screening of microalgae growth potential.

    PubMed

    Van Wagenen, Jon; Holdt, Susan Løvstad; De Francisci, Davide; Valverde-Pérez, Borja; Plósz, Benedek Gy; Angelidaki, Irini

    2014-10-01

    Microalgae cultivation conditions in microplates will differ from large-scale photobioreactors in crucial parameters such as light profile, mixing and gas transfer. Hence volumetric productivity (P(v)) measurements made in microplates cannot be directly scaled up. Here we demonstrate that it is possible to use microplates to measure characteristic exponential growth rates and determine the specific growth rate light intensity dependency (μ-I curve), which is useful as the key input for several models that predict P(v). Nannochloropsis salina and Chlorella sorokiniana specific growth rates were measured by repeated batch culture in microplates supplied with continuous light at different intensities. Exponential growth unlimited by gas transfer or self-shading was observable for a period of several days using fluorescence, which is an order of magnitude more sensitive than optical density. The microplate datasets were comparable to similar datasets obtained in photobioreactors and were used an input for the Huesemann model to accurately predict P(v).

  20. Tectonics and evolution of the Juan Fernandez microplate at the Pacific-Nazca-Antarctic triple junction

    NASA Technical Reports Server (NTRS)

    Anderson-Fontana, S.; Larson, R. L.; Engein, J. F.; Lundgren, P.; Stein, S.

    1986-01-01

    Magnetic and bathymetric profiles derived from the R/V Endeavor survey and focal mechanism studies for earthquakes on two of the Juan Fernandez microplate boundaries are analyzed. It is observed that the Nazca-Juan Fernandez pole is in the northern end of the microplate since the magnetic lineation along the East Ridge of the microplate fans to the south. The calculation of the relative motion of the Juan Fernandez-Pacific-Nazca-Antarctic four-plate system using the algorithm of Minster et al. (1974) is described. The development of tectonic and evolutionary models of the region is examined. The tectonic model reveals that the northern boundary of the Juan Fernandez microplate is a zone of compression and that the West Ridge and southwestern boundary are spreading obliquely; the evolutionary model relates the formation of the Juan Fernandez microplate to differential spreading rates at the triple junction.

  1. Understanding Readers' Differing Understandings

    ERIC Educational Resources Information Center

    Kucer, Stephen B.

    2015-01-01

    This research examines the characteristics of reader understandings that vary from those stated in the text. Eighty-seven fourth graders orally read complex academic literary and scientific texts, followed by probed retellings. Retold ideas not directly supported by, or reflective of, the texts were identified. These differing understandings…

  2. Hindi Basic Reader.

    ERIC Educational Resources Information Center

    Harter, J. Martin; And Others

    This reader is intended to accompany the Basic Course in Spoken Hindi. Following an outline of the Devanagari script, 20 lessons are presented. Each consists of a reading selection, several illustrative sentences in English and Hindi, and a series of questions. Most of the reading selections were adapted from the magazine "Bal-Bharati."…

  3. Advanced Tagalog Reader.

    ERIC Educational Resources Information Center

    Ramos, Teresita V.

    A Tagalog reader designed for adult students beyond the intermediate level contains 30 lessons in agriculture, art, economics, education, history, language, literature, medicine, music, political science, religion, and sociology. Each unit contains four sections: (1) a reading passage with numbered paragraphs and underlined vocabulary words, for…

  4. Growing Young Gifted Readers

    ERIC Educational Resources Information Center

    Wood, Patricia F.

    2009-01-01

    There is great pressure on parents to teach children to read as early as possible. In reality, precocious (early advanced) readers seem to almost master the skill on their own, without the assistance of highly touted, commercially available programs. The 18-month-old toddler who names the letters on alphabet blocks; or the 26-month-old who can…

  5. Comparison of turbulent-flow chromatography with automated solid-phase extraction in 96-well plates and liquid-liquid extraction used as plasma sample preparation techniques for liquid chromatography-tandem mass spectrometry.

    PubMed

    Zimmer, D; Pickard, V; Czembor, W; Müller, C

    1999-08-27

    Turbulent flow chromatography (TFC) combined with the high selectivity and sensitivity of tandem mass spectrometry (MS-MS) is a new technique for the fast direct analysis of drugs from crude plasma. TFC in the 96-well plate format reduces significantly the time required for sample clean-up in the laboratory. For example, for 100 samples the workload for a technician is reduced from about 8 h by a manual liquid-liquid extraction (LLE) assay to about 1 h in the case of TFC. Sample clean-up and analysis are performed on-line on the same column. Similar chromatographic performance and validation results were achieved using HTLC Turbo-C18 columns (Cohesive Technologies) and Oasis HLB extraction columns (Waters). One 96-well plate with 96 plasma samples is analyzed within 5.25 h, corresponding to 3.3 min per sample. Compared to this LLE and analysis of 96 samples takes about 16 h. Two structurally different and highly protein bound compounds, drug A and drug B, were analyzed under identical TFC conditions and the assays were fully validated for the application to toxicokinetics studies (compliant with Good Laboratory Practices-GLP). The limit of quantitation was 1.00 microg/l and the linear working range covered three orders of magnitude for both drugs. In the case of drug A the quality of analysis by TFC was similar to the reference LLE assay and slightly better than automated solid-phase extraction in 96-well plates. The accuracy was -3.1 to 6.7% and the precision was 3.1 to 6.8% in the case of drug A determined for dog plasma by TFC-MS-MS. For drug B the accuracy was -3.7 to 3.5% and the precision was 1.6 to 5.4% for rat plasma, which is even slightly better than what was achieved with the validated protein precipitation assay.

  6. NaturalReader: A New Generation Text Reader

    ERIC Educational Resources Information Center

    Flood, Jacqueline

    2007-01-01

    NaturalReader (http://www.naturalreaders.com/) is a new generation text reader, which means that it reads any machine readable text using synthesized speech without having to copy and paste the selected text into the NaturalReader application window. It installs a toolbar directly into all of the Microsoft Office[TM] programs and uses a mini-board…

  7. Readers' Readings: Applications of Reader-Response Theory.

    ERIC Educational Resources Information Center

    Steiner, Linda

    In the interest of applying reader response theory to journalism this paper posits that readers of newspapers, like readers of literature, take an active role in making meaning from the articles they read, rather than passively accepting news as a finished, static product. Additionally, it proposes that journalism textbooks pay little attention to…

  8. Liquid chromatography/tandem mass spectrometric bioanalysis using normal-phase columns with aqueous/organic mobile phases - a novel approach of eliminating evaporation and reconstitution steps in 96-well SPE.

    PubMed

    Naidong, Weng; Shou, Wilson Z; Addison, Thomas; Maleki, Saber; Jiang, Xiangyu

    2002-01-01

    Bioanalytical methods using automated 96-well solid-phase extraction (SPE) and liquid chromatography with electrospray tandem mass spectrometry (LC/MS/MS) are widely used in the pharmaceutical industry. SPE methods typically require manual steps of drying of the eluates and reconstituting of the analytes with a suitable injection solvent possessing elution strength weaker than the mobile phase. In this study, we demonstrated a novel approach of eliminating these two steps in 96-well SPE by using normal-phase LC/MS/MS methods with low aqueous/high organic mobile phases, which consisted of 70-95% organic solvent, 5-30% water, and small amount of volatile acid or buffer. While the commonly used SPE elution solvents (i.e. acetonitrile and methanol) have stronger elution strength than a mobile phase on reversed-phase chromatography, they are weaker elution solvents than a mobile phase for normal-phase LC/MS/MS and therefore can be injected directly. Analytical methods for a range of polar pharmaceutical compounds, namely, omeprazole, metoprolol, fexofenadine, pseudoephedrine as well as rifampin and its metabolite 25-desacetyl-rifampin, in biological fluids, were developed and optimized based on the foregoing principles. As a result of the time saving, a batch of 96 samples could be processed in one hour. These bioanalytical LC/MS/MS methods were validated according to "Guidance for Industry - Bioanalytical Method Validation" recommended by the Food and Drug Administration (FDA) of the United States.

  9. β-Cyclodextrin enhanced on-line organic solvent field-amplified sample stacking in capillary zone electrophoresis for analysis of ambroxol in human plasma, following liquid-liquid extraction in the 96-well format.

    PubMed

    Li, Ji; Bi, Youwei; Wang, Li; Sun, Fanlu; Chen, Zhao; Xu, Guili; Fan, Guorong

    2012-07-01

    A field-amplified sample stacking (FASS) and capillary zone electrophoresis (CZE) method is described for the quantification of ambroxol hydrochloride in human plasma, following liquid-liquid extraction in the 96-well format. The separation was carried out at 25 °C in a 31.2 cm × 75 μm fused-silica capillary with an applied voltage of 15 kV. The background electrolyte (BGE) was composed of 6.25 mM borate-25 mM phosphate (pH 3.0) and 1mM β-cyclodextrin. The detection wavelength was 210 nm. Clean-up and preconcentration of plasma biosamples were developed by 96-well format liquid-liquid extraction (LLE). In this study, FASS in combination with β-cyclodextrin enhanced the sensitivity about 60-70 fold in total. The method was suitably validated with respect to stability, specificity, linearity, lower limit of quantitation, accuracy, precision, extraction recovery and robustness. The calibration graph was linear for ambroxol hydrochloride from 2 to 500 ng/ml. The lower limit of quantification was 2 ng/ml. The intra- and inter-day precisions of lowest limit of quantification (LLOQ) were 9.61 and 11.80%, respectively. The method developed was successfully applied to the evaluation of clinical pharmacokinetic study of ambroxol hydrochloride tablet after oral administration to 12 healthy volunteers.

  10. The western transverse ranges microplate as a native terrane

    SciTech Connect

    Campbell, M.D.; Reed, W.E. )

    1994-04-01

    Palocurrent measurements from the entire Cretaceous section of the western Transverse Ranges microplate (WTRM) yield a northerly flow direction. Point count data indicate a mixed provenance for both conglomerates and associated sandstones. The dominant provenance was mixed magmatic arc/recycled orogen and disected/transitional arc terranes. Petrographic, quantitative SEM and microprobe analysis also indicate the presence of diagnostic Franciscan mineralogy in these sediments, including glaucophane, riebeckite, lawsonite, and serpentine, suggesting derivation from a subduction complex. Olistoclasts of chert, jadeitic graywacke, serpentine and blueschist are found intermixed within the arc-derived sediments. Olistoclasts range in size from sub-millimeter to centimeter scale and olistoliths range up to 150 m. Well preserved internal bedding in some of the olistoliths suggest emplacement by landsliding indicating very short transport distance. This Franciscan material represents the oldest melange-derived material reported from this part of California and documents uplift and erosion of the subduction complex earlier than previously suggested. These data are consistent with deposition in a Cretaceous fore-arc basin located west or south of the San Diego area. The allochthonous WTRM of southern California can be reconstructed to an originally north-south oriented fore-arc basin. After deposition of the Sespe Formation (22 Ma [+-]) the microplate was slivered by strike-slip faults and rotated clockwise approximately 90[degrees], after which, the block again accreted against the continental margin. Our reconstruction suggest that depositional and structural trends for Eocene and Cretaceous sediments is likely to be different from that in the Miocene Monterey pay zones in the Santa Barbara channel region. If our reconstruction is correct, exploration strategy for Eocene and Cretaceous petroleum in the southern California Bight should take this tectonic model into account.

  11. Readers of histone modifications

    PubMed Central

    Yun, Miyong; Wu, Jun; Workman, Jerry L; Li, Bing

    2011-01-01

    Histone modifications not only play important roles in regulating chromatin structure and nuclear processes but also can be passed to daughter cells as epigenetic marks. Accumulating evidence suggests that the key function of histone modifications is to signal for recruitment or activity of downstream effectors. Here, we discuss the latest discovery of histone-modification readers and how the modification language is interpreted. PMID:21423274

  12. My Journey as a Reader

    ERIC Educational Resources Information Center

    Cevellos, Tatiana

    2008-01-01

    In this case history, the author describes how her journey as a reader evolved from a poor reader who did not like to read in elementary school into an avid trilingual reader in graduate school. Once she discovered the joy of reading, each language in which she read had its own purpose and emotional connection. She credits self-selected reading…

  13. Rapid screening of purification strategies for the capture of a human recombinant F(ab')2 expressed in baculovirus-infected cells using a micro-plate approach and SELDI-MS.

    PubMed

    Pezzini, J; Brenac Brochier, V; Barrouillet, M P; Cerruti, M; Clofent-Sanchez, G; Schapman, A; Topol, A; Robert, R; Cabanne, C; Cerruti, P; Santarelli, X

    2009-08-15

    The development of a capture step of a human recombinant F(ab')(2) produced and expressed in baculovirus-infected cells was investigated by screening three mixed-mode chromatography sorbents (HEA HyperCel, PPA HyperCel and MEP HyperCel) and two ion exchangers (Q Ceramic HyperD F, S Ceramic HyperD F) sorbents using a 96-well plate format and SELDI-MS. HEA HyperCel gave the best separation performance therefore the conditions tested in micro-plate were transferred to laboratory scale chromatographic experiments, confirming that the recombinant F(ab')(2) was effectively captured on the mixed-mode sorbent without any pre-treatment of the crude extract with a 82% recovery and a 39-fold purification.

  14. A fluorescent microplate assay quantifies bacterial efflux and demonstrates two distinct compound binding sites in AcrB.

    PubMed

    Iyer, Ramkumar; Ferrari, Annette; Rijnbrand, R; Erwin, Alice L

    2015-04-01

    A direct assay of efflux by Escherichia coli AcrAB-TolC and related multidrug pumps would have great value in discovery of new Gram-negative antibiotics. The current understanding of how efflux is affected by the chemical structure and physical properties of molecules is extremely limited, derived from antibacterial data for compounds that inhibit growth of wild-type E. coli. We adapted a previously described fluorescent efflux assay to a 96-well microplate format that measured the ability of test compounds to compete for efflux with Nile Red (an environment-sensitive fluor), independent of antibacterial activity. We show that Nile Red and the lipid-sensitive probe DiBAC4-(3) [bis-(1,3-dibutylbarbituric acid)-trimethine oxonol] can quantify efflux competition in E. coli. We extend the previous findings that the tetracyclines compete with Nile Red and show that DiBAC4-(3) competes with macrolides. The extent of the competition shows a modest correlation with the effect of the acrB deletion on MICs within the compound sets for both dyes. Crystallographic studies identified at least two substrate binding sites in AcrB, the proximal and distal pockets. High-molecular-mass substrates bound the proximal pocket, while low-mass substrates occupied the distal pocket. As DiBAC4-(3) competes with macrolides but not with Nile Red, we propose that DiBAC4-(3) binds the proximal pocket and Nile Red likely binds the distal site. In conclusion, competition with fluorescent probes can be used to study the efflux process for diverse chemical structures and may provide information as to the site of binding and, in some cases, enable rank-ordering a series of related compounds by efflux.

  15. A Fluorescent Microplate Assay Quantifies Bacterial Efflux and Demonstrates Two Distinct Compound Binding Sites in AcrB

    PubMed Central

    Ferrari, Annette; Rijnbrand, R.; Erwin, Alice L.

    2015-01-01

    A direct assay of efflux by Escherichia coli AcrAB-TolC and related multidrug pumps would have great value in discovery of new Gram-negative antibiotics. The current understanding of how efflux is affected by the chemical structure and physical properties of molecules is extremely limited, derived from antibacterial data for compounds that inhibit growth of wild-type E. coli. We adapted a previously described fluorescent efflux assay to a 96-well microplate format that measured the ability of test compounds to compete for efflux with Nile Red (an environment-sensitive fluor), independent of antibacterial activity. We show that Nile Red and the lipid-sensitive probe DiBAC4-(3) [bis-(1,3-dibutylbarbituric acid)-trimethine oxonol] can quantify efflux competition in E. coli. We extend the previous findings that the tetracyclines compete with Nile Red and show that DiBAC4-(3) competes with macrolides. The extent of the competition shows a modest correlation with the effect of the acrB deletion on MICs within the compound sets for both dyes. Crystallographic studies identified at least two substrate binding sites in AcrB, the proximal and distal pockets. High-molecular-mass substrates bound the proximal pocket, while low-mass substrates occupied the distal pocket. As DiBAC4-(3) competes with macrolides but not with Nile Red, we propose that DiBAC4-(3) binds the proximal pocket and Nile Red likely binds the distal site. In conclusion, competition with fluorescent probes can be used to study the efflux process for diverse chemical structures and may provide information as to the site of binding and, in some cases, enable rank-ordering a series of related compounds by efflux. PMID:25645845

  16. Fluorescence microplate readers as an alternative to flow injection analysis for determination of wort beta-glucan

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Selection of new barley varieties to meet the needs of the malting and brewing industry requires that the lines being developed meet a set of malting quality standards that have been developed to help maltsters and brewers predict the commercial performance of the barley and resulting malt. One imp...

  17. Fluorescence microplate readers as an alternative to flow injection analysis for determination of wort beta-glucan

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wort beta-glucan concentration is a critical malting quality parameter used to identify and avoid potential brewhouse filtration problems. ASBC method Wort-18 is widely used in malt analysis laboratories and brewhouses to measure wort beta-glucan levels. However, the chemistry underlying the method...

  18. A simple 96-well liquid-liquid extraction with a mixture of acetonitrile and methyl t-butyl ether for the determination of a drug in human plasma by high-performance liquid chromatography with tandem mass spectrometry.

    PubMed

    Xue, Y-J; Pursley, Janice; Arnold, Mark E

    2004-02-04

    A simple 96-well plate liquid-liquid extraction (LLE), liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the determination of a basic drug candidate in human plasma. Against the wisdom of conventional approaches, an aqueous/organic miscible solvent, acetonitrile, was used for liquid-liquid extraction along with methyl t-butyl ether. The use of acetonitrile effectively eliminated the formation of the irregular emulsion between aqueous/organic interfaces and modulated the polarity of the extraction solvents to achieve the desired recovery. This approach, which solved the emulsion problem, permitted the method to be automated using standard 96-well plate technology. A practical application was demonstrated through the use of this technique in the measurement of a novel drug in human plasma samples by LC/MS/MS. Chromatographic separation was achieved isocratically on a Phenomenox C18(2) Luna column (2 mm x 50 mm, 5 microm). The mobile phase contained 60% of 0.1% formic acid and 40% acetonitrile. Detection was by positive ion electrospray tandem mass spectrometry. The standard curve, which ranged from 1.22 to 979ng/ml, was fitted to a 1/x2 weighted quadratic regression model. The validation results show that this method was very rugged and had excellent precision and accuracy. The actual sample analysis results further demonstrated that this extraction procedure is well suited for real life applications. It is expected that with some modifications, this approach can be applied for the extraction of similar compounds from various biological fluids.

  19. A 96-well single-pot protein precipitation, liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the determination of muraglitazar, a novel diabetes drug, in human plasma.

    PubMed

    Xue, Y-J; Liu, Jane; Pursley, Janice; Unger, Steve

    2006-02-02

    A 96-well single-pot protein precipitation, liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the determination of muraglitazar, a PPAR alpha/gamma dual agonist, in human plasma. The internal standard, a chemical analogue, was dissolved in acetonitrile containing 0.1% formic acid. The solvent system was also served as a protein precipitation reagent. Human plasma samples (0.1 mL) and the internal standard solution (0.3 mL) were added to a 96-well plate. The plate was vortexed for 1 min and centrifuged for 5 min. Then the supernatant layers were directly injected into the LC/MS/MS system. The chromatographic separation was achieved isocratically on a Phenomenox C18(2) Luna column (2 mm x 50 mm, 5 microm). The mobile phase contained 20/80 (v/v) of water and acetonitrile containing 0.1% formic acid. Detection was by positive ion electrospray tandem mass spectrometry on a Sciex API 3000. The standard curve, which ranged from 1 to 1000 ng/mL, was fitted to a 1/x weighted quadratic regression model. This single-pot approach effectively eliminated three time consuming sample preparation steps: sample transfer, dry-down, and reconstitution before the injection, while it preserved all the benefits of the traditional protein precipitation. By properly adjusting the autosampler needle offset level, only the supernatant was injected, without disturbing the precipitated proteins in the bottom. As a result, the quality of chromatography and column life were not compromised. After more than 600 injections, there was only slightly increase of column back-pressure. The validation results demonstrated that this method was rugged and provide satisfactory precision and accuracy. The method has been successfully applied to analyze human plasma samples in support of a first-in-man study. This method has also been validated in monkey and mouse plasma for the determination of muraglitazar.

  20. High-throughput sample preparation for the quantitation of acephate, methamidophos, omethoate, dimethoate, ethylenethiourea, and propylenethiourea in human urine using 96-well-plate automated extraction and high-performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Jayatilaka, Nayana K; Angela Montesano, M; Whitehead, Ralph D; Schloth, Sara J; Needham, Larry L; Barr, Dana Boyd

    2011-07-01

    Acephate, methamidophos, o-methoate, and dimethoate are organophosphorus pesticides, and ethylenethiouria and propylenethiourea are two metabolites from the bisdithiocarbamate fungicide family. They are some of the most widely used pesticides and fungicides in agriculture both domestically and abroad. The existing high-performance liquid chromatography (HPLC)-tandem mass spectrometry (MS/MS) method for the measurement of these compounds in human urine was improved by using a 96-well plate format sample preparation; the use of HPLC-MS/MS was comparable with a concentration range of 0.125 to 50 ng/ml. Deuterium-labeled acephate, ethylenethiouria, and methamidophos were used as internal standards. The sample preparation procedure, in the 96-well format with a 0.8-ml urine sample size, uses lyophilization of samples, followed by extraction with dichloromethane. The analytes were chromatographed on a Zorbax SB-C3 (4.6 × 150 mm, 5.0-μm) column with gradient elution by using 0.1% formic acid in aqueous solution (solvent A) and 0.1% formic acid in methanol (solvent B) mobile phase at a flow rate of 1 ml/min. Quantitative analysis was performed by atmospheric pressure chemical ionization source in positive ion mode using multiple-reaction monitoring of the precursor-to-product ion pairs for the analytes on a TSQ Quantum Ultra HPLC-MS/MS. Repeated analyses of urine samples spiked with high (15 ng/ml), medium (5 ng/ml), and low (1 ng/ml) concentrations of the analytes gave relative SDs of <13%. The limits of detection were in the range of 0.004-0.01 ng/ml. The method also has high accuracy, high precision, and excellent extraction recovery. Furthermore, the improved sample preparation method decreased the cost and labor required while effectively doubling the analytic throughput with minimal matrix effect.

  1. Pull-in instability tuning in imperfect nonlinear circular microplates under electrostatic actuation

    NASA Astrophysics Data System (ADS)

    Jallouli, A.; Kacem, N.; Bourbon, G.; Le Moal, P.; Walter, V.; Lardies, J.

    2016-12-01

    Dynamic range improvement based on geometric nonlinearity and initial deflection is demonstrated with imperfect circular microplates under electrostatic actuation. Depending on design parameters, we prove how the von Kármán nonlinearity and the plate imperfections lead to a significant delay in pull-in occurrence. These promising results open the way towards an accurate identification of static parameters of circular microplates and the development of a predictive model for the nonlinear dynamics of imperfect capacitive micromachined ultrasonic transducers.

  2. The larval parasitoid Microplitis croceipes oviposits in conspecific adults

    NASA Astrophysics Data System (ADS)

    Takasu, Keiji; Hoang Le, K.

    2007-03-01

    Microplitis croceipes (Hymenoptera: Braconidae) is a larval parasitoid of Helicoverpa/Heliothis spp. In the course of mass rearing of M. croceipes, we found that females oviposited in the conspecific adults in rearing cages. When 20 pairs of inexperienced females and males or of experienced females and males were reared in a cage, the males lived for 14-15 days and the females for 18-20 days on average. At their death, 37-42% of the males and 50-57% of the females contained conspecific eggs or first instar larvae in their abdominal cavity. When two of inexperienced females met on a host-infested leaf of soybean, they attempted to sting each other. Of the attacked females, 30% contained a conspecific egg laid in their abdomen. In abdominal cavity of the adults parasitized by a conspecific female, the majority of the parasitoid eggs laid disappeared within 1 day after oviposition. Only 10-30% of the parasitoid eggs laid in conspecific adults hatched 3-4 days after oviposition, but those larvae never molted to second instar. When the adults were stung by one or two conspecific females, their subsequent longevity was significantly shorter than that for the control adults. Oviposition in conspecific adults may be prevalent in other parasitic wasps that quickly oviposit without intensive host examination, and have cuticle and size of abdomen to be stung by conspeicifcs.

  3. Development and application of a high-throughput sample cleanup process based on 96-well plate for simultaneous determination of 16 steroids in biological matrices using liquid chromatography-triple quadrupole mass spectrometry.

    PubMed

    Luo, Guanzhong; Li, Youxin; Bao, James J

    2016-02-01

    A novel high-throughput sample pretreatment system was developed by the integration of protein precipitation (PP), phospholipid removal (PPR), and hollow fiber liquid-phase microextraction (HF-LPME) into two simple 96-well plates and a matching 96-grid lid. With this system, 16 steroids were separated from biological matrices of plasma, milk, and urine and analyzed by liquid chromatography-triple quadrupole mass spectrometry. In the tandem sample cleanup process, the prepositive PP and PPR step preliminarily removed some of the interferences from the biological matrices. The following HF-LPME step kept the residual interference out of the hollow fiber and enriched the steroids in the hollow fiber to achieve high sensitivity. By a series of method optimizations, acetonitrile was chosen as the crash solvent for PP and PPR. A mixture of octanol and toluene (1:1 v/v) was used as the acceptor phase for HF-LPME. The extraction was conducted at 80 rpm for 50 min in a donor phase containing 1 mL 20% sodium chloride at 25 °C. Under these conditions, the limits of detection for the 16 steroids were 3.6-300.0 pg(.)mL(-1) in plasma, 3.0-270.0 pg·mL(-1) in milk, and 2.2-210.0 pg(.)mL(-1) in urine. The recoveries of the 16 steroids were 81.9-97.9% in plasma (relative standard deviation 1.0-8.0%), 80.6-97.7% in milk (relative standard deviation 0.8-5.4%), and 87.3-98.7% in urine (relative standard deviation 1.0-4.9%). Further, the integrated 96-well platform of PP, PPR, and HF-LPME enabled us to run this assay in an automatic and high-throughput fashion. The reliability of the method was further corroborated by evaluation of its applicability in plasma and urine samples from volunteers and fresh bovine milk from local dairy enterprises.

  4. Automated 96-well solid phase extraction and hydrophilic interaction liquid chromatography-tandem mass spectrometric method for the analysis of cetirizine (ZYRTEC) in human plasma--with emphasis on method ruggedness.

    PubMed

    Song, Qi; Junga, Heiko; Tang, Yong; Li, Austin C; Addison, Tom; McCort-Tipton, Melanie; Beato, Brian; Naidong, Weng

    2005-01-05

    A high-throughput bioanalytical method based on automated sample transfer, automated solid phase extraction, and hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) analysis, has been developed for the determination of cetirizine, a selective H(1)-receptor antagonist. Deuterated cetirizine (cetirizine-d(8)) was synthesized as described and was used as the internal standard. Samples were transferred into 96-well plates using an automated sample handling system. Automated solid phase extraction was carried out using a 96-channel programmable liquid-handling workstation. Solid phase extraction 96-well plate on polymer sorbent (Strata X) was used to extract the analyte. The extracted samples were injected onto a Betasil silica column (50 x 3, 5 microm) using a mobile phase of acetonitrile-water-acetic acid-trifluroacetic acid (93:7:1:0.025, v/v/v/v) at a flow rate of 0.5 ml/min. The chromatographic run time is 2.0 min per injection, with retention time of cetirizine and cetirizine-d(8) both at 1.1 min. The system consisted of a Shimadzu HPLC system and a PE Sciex API 3000 or API 4000 tandem mass spectrometer with (+) ESI. The method has been validated over the concentration range of 1.00-1000 ng/ml cetirizine in human plasma, based on a 0.10-ml sample size. The inter-day precision and accuracy of the quality control (QC) samples demonstrated <3.0% relative standard deviation (R.S.D.) and <6.0% relative error (RE). Stability of cetirizine in stock solution, in plasma, and in reconstitution solution was established. The absolute extraction recovery was 85.8%, 84.5%, and 88.0% at 3, 40, and 800 ng/ml, respectively. The recovery for the internal standard was 84.1%. No adverse matrix effects were noticed for this assay. The automation of the sample preparation steps not only increased the analysis throughput, but also increased method ruggedness. The use of a stable isotope-labeled internal standard further improved the method ruggedness

  5. The Graded Reader Is Dead, Long Live the Electronic Reader

    ERIC Educational Resources Information Center

    Eldridge, John; Neufeld, Steven

    2009-01-01

    Many course books for learners of English as a second or foreign language now claim to contain a strong lexical component. Other practitioners meanwhile continue to advocate the use of graded readers to augment vocabulary development. This article reports the findings of vocabulary profiling and analysis of both course books and graded readers and…

  6. A rapid and high-throughput microplate spectrophotometric method for field measurement of nitrate in seawater and freshwater.

    PubMed

    Wu, Jiapeng; Hong, Yiguo; Guan, Fengjie; Wang, Yan; Tan, Yehui; Yue, Weizhong; Wu, Meilin; Bin, Liying; Wang, Jiaping; Wen, Jiali

    2016-02-01

    The well-known zinc-cadmium reduction method is frequently used for determination of nitrate. However, this method is seldom to be applied on field research of nitrate due to the long time consuming and large sample volume demand. Here, we reported a modified zinc-cadmium reduction method (MZCRM) for measurement of nitrate at natural-abundance level in both seawater and freshwater. The main improvements of MZCRM include using small volume disposable tubes for reaction, a vortex apparatus for shaking to increase reduction rate, and a microplate reader for high-throughput spectrophotometric measurements. Considering salt effect, two salinity sections (5~10 psu and 20~35 psu) were set up for more accurate determination of nitrate in low and high salinity condition respectively. Under optimized experimental conditions, the reduction rates were stabilized on 72% and 63% on the salinity of 5 and 20 psu respectively. The lowest detection limit for nitrate was 0.5 μM and was linear up to 100 μM (RSDs was 4.8%). Environmental samples assay demonstrated that MZCRM was well consistent with conventional zinc-cadmium reduction method. In total, this modified method improved accuracy and efficiency of operations greatly, and would be realized a rapid and high-throughput determination of nitrate in field analysis of nitrate with low cost.

  7. Automated liquid-liquid extraction based on 96-well plate format in conjunction with ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for the quantitation of methoxsalen in human plasma.

    PubMed

    Yadav, Manish; Contractor, Pritesh; Upadhyay, Vivek; Gupta, Ajay; Guttikar, Swati; Singhal, Puran; Goswami, Sailendra; Shrivastav, Pranav S

    2008-09-01

    A sensitive, specific and high throughput bioanalytical method using automated sample processing via 96-well plate liquid-liquid extraction and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) has been developed for the determination of methoxsalen in human plasma. Plasma samples with ketoconazole as internal standard (IS) were prepared by employing 0.2 mL human plasma in ethyl acetate:dichloromethane (80:20, v/v). The chromatographic separation was achieved on a Waters Acquity UPLC BEH C18 column using isocratic mobile phase, consisting of 10 mM ammonium formate and acetonitrile (60:40, v/v), at a flow rate of 0.5 mL/min. The linear dynamic range was established over the concentration range 1.1-213.1 ng/mL for methoxsalen. The method was rugged and rapid with a total run time of 1.5 min. It was successfully applied to a pivotal bioequivalence study in 12 healthy human subjects after oral administration of 10 mg extended release methoxsalen formulation under fasting condition.

  8. Native American Adult Reader I.

    ERIC Educational Resources Information Center

    King, Lovern Root, Ed.

    Aspects of Native American history and culture as well as issues and concerns of American Indians are presented in the twelve short articles in this reader for adults. Intended for use in an adult basic education/GED program, the reader features simply written stories (for grades 0-3), illustrations, vocabulary lists and student study questions.…

  9. Readers, Instruction, and the NRP

    ERIC Educational Resources Information Center

    Wilson, G. Pat; Martens, Prisca; Arya, Poonam; Altwerger, Bess

    2004-01-01

    Are programs that emphasize systematic phonics instruction truly superior to other types of programs for young readers, as the National Reading Panel claims? The authors conducted a study of three different programs to see what kinds of readers are actually emerging from them. Two were commercial programs that used explicit and systematic phonics…

  10. Proud Readers Own What's Learned

    ERIC Educational Resources Information Center

    Tucker, Jessica Fink

    2014-01-01

    Proud Readers Own What's Learned (PROWL) was developed out of the need to find an engaging, creative process for readers to express what they have read and, frankly, to let teachers see that the students actually read the books. In other more prescriptive reading programs students can go through the motions and actually pass tests without having…

  11. TESOL Reader and Teachers Guide.

    ERIC Educational Resources Information Center

    Hanson-Smith, Elizabeth

    This combined reader and teacher's guide for English second language instruction was developed from teacher workshops conducted in Belize. The reader consists of stories, poems, and drama from the indigenous people of that country, accompanied by supporting English language learning exercises and activities. Texts represent aspects of indigenous…

  12. The Routledge Applied Linguistics Reader

    ERIC Educational Resources Information Center

    Wei, Li, Ed.

    2011-01-01

    "The Routledge Applied Linguistics Reader" is an essential collection of readings for students of Applied Linguistics. Divided into five sections: Language Teaching and Learning, Second Language Acquisition, Applied Linguistics, Identity and Power and Language Use in Professional Contexts, the "Reader" takes a broad…

  13. Readers Responding--And Then?

    ERIC Educational Resources Information Center

    Hansson, Gunnar

    1992-01-01

    Argues that recognition of the reader's role in the production of meanings in literary works has major consequences for the theory, research, and teaching of literature. Discusses the need to include the reader's role when writing the history of literature; develop new language for the description of the meanings of works; and develop new models…

  14. Magmatic evolution of the Easter microplate-Crough Seamount region (South East Pacific)

    USGS Publications Warehouse

    Hekinian, R.; Stoffers, P.; Akermand, D.; Binard, N.; Francheteau, Jean; Devey, C.; Garbe-Schonberg, D.

    1995-01-01

    The Easter microplate-Crough Seamount region located between 25?? S-116?? W and 25?? S-122?? W consists of a chain of seamounts forming isolated volcanoes and elongated (100-200 km in length) en echelon volcanic ridges oriented obliquely NE (N 065??), to the present day general spreading direction (N 100??) of the Pacific-Nazca plates. The extension of this seamount chain into the southwestern edge of the Easter microplate near 26??30??? S-115?? W was surveyed and sampled. The southern boundary including the Orongo fracture zone and other shallow ridges ( 0.25) MORBs which are similar in composition to other more recent basalts from the Southwest and East Rifts spreading axes of the Easter microplate. Incompatible element ratios normalized to chondrite values [(Ce/Yb)N = 1-2.5}, {(La/Sm)N = 0.4-1.2} and {(Zr/Y)N = 0.7-2.5} of the basalts are also similar to present day volcanism found in the Easter microplate. The volcanics from the Easter microplate-Crough region are unrelated to other known South Pacific intraplate magmatism (i.e. Society, Pitcairn, and Salas y Gomez Islands). Instead their range in incompatible element ratios is comparable to the submarine basalts from the recently investigated Ahu and Umu volcanic field (Easter hotspot) (Scientific Party SO80, 1993) and centered at about 80 km west of Easter Island. The oblique ridges and their associated seamounts are likely to represent ancient leaky transform faults created during the initial stage of the Easter microplate formation (??? 5 Ma). It appears that volcanic activity on seamounts overlying the oblique volcanic ridges has continued during their westward drift from the microplate as shown by the presence of relatively fresh lava observed on one of these structures, namely the first Oblique Volcanic Ridge near 25?? S-118?? W at about 160 km west of the Easter microplate West Rift. Based on a reconstruction of the Easter microplate, it is suggested that the Crough seamount (< 800 m depth) was formed

  15. Work function engineering of SnO single crystal microplates with thermal annealing.

    PubMed

    Doh, Won Hui; Jeong, Wooseok; Lee, Hyunsoo; Park, Jonghyurk; Park, Jeong Young

    2016-08-19

    We synthesized black SnO single-crystal microplates via a sonochemical process and engineered the work function of the SnO microplates using thermal treatments. The as-synthesized SnO microplates have a wide (001) plane, as is clearly evident from TEM images and diffraction patterns. Surface potential measurements on the SnO microplates show that the work function changes as the annealing temperature increases. The TEM and XAS results after thermal treatments imply that the micro-sized SnO(001) single-crystals are stable up to about 400 °C in air, after which the surface starts to become locally oxidized. Consequently, the long-range ordering and lattice parameter of the SnO(001) single crystals started to change to make polycrystalline SnO2 at about 600 °C. These results demonstrate the ability to tune the work function of the microplates and suggest an intriguing way to engineer the electrical properties of nanostructures.

  16. Validation of a sensitive and automated 96-well solid-phase extraction liquid chromatography-tandem mass spectrometry method for the determination of desloratadine and 3-hydroxydesloratadine in human plasma.

    PubMed

    Yang, Liyu; Clement, Robert P; Kantesaria, Bhavna; Reyderman, Larisa; Beaudry, Francis; Grandmaison, Charles; Di Donato, Lorella; Masse, Robert; Rudewicz, Patrick J

    2003-07-25

    To support clinical development, a liquid chromatographic-tandem mass spectrometric (LC-MS-MS) method was developed and validated for the determination of desloratadine (descarboethoxyloratadine) and 3-OH desloratadine (3-hydroxydescarboethoxyloratadine) concentrations in human plasma. The method consisted of automated 96-well solid-phase extraction for sample preparation and liquid chromatography/turbo ionspray tandem mass spectrometry for analysis. [2H(4)]Desloratadine and [2H(4)]3-OH desloratadine were used as internal standards (I.S.). A quadratic regression (weighted 1/concentration(2)) gave the best fit for calibration curves over the concentration range of 25-10000 pg/ml for both desloratadine and 3-OH desloratadine. There was no interference from endogenous components in the blank plasma tested. The accuracy (%bias) at the lower limit of quantitation (LLOQ) was -12.8 and +3.4% for desloratadine and 3-OH desloratadine, respectively. The precision (%CV) for samples at the LLOQ was 15.1 and 10.9% for desloratadine and 3-OH desloratadine, respectively. For quality control samples at 75, 1000 and 7500 pg/ml, the between run %CV was

  17. Validation and application of a liquid chromatography-tandem mass spectrometric method for the determination of GDC-0834 and its metabolite in human plasma using semi-automated 96-well protein precipitation.

    PubMed

    Shin, Young G; Jones, Steve A; Murakami, Stan C; Liu, Lichuan; Wong, Harvey; Buonarati, Michael H; Hop, Cornelis E C A

    2012-11-01

    A liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method was developed and validated for the determination of GDC-0834 and its amide hydrolysis metabolite (M1) in human plasma to support clinical development. The method consisted of semi-automated 96-well protein precipitation extraction for sample preparation and LC-MS/MS analysis in positive ion mode using TurboIonSpray® for analysis. D6-GDC-0834 and D6-M1 metabolite were used as internal standards. A linear regression (weighted 1/concentration(2) ) was used to fit calibration curves over the concentration range of 1 - 500 ng/mL for both GDC-0834 and M1 metabolite. The accuracy (percentage bias) at the lower limit of quantitation (LLOQ) was 5.20 and 0.100% for GDC-0834 and M1 metabolite, respectively. The precision (CV) for samples at the LLOQ was 3.13-8.84 and 5.20-8.93% for GDC-0834 and M1 metabolite, respectively. For quality control samples at 3, 200 and 400 ng/mL, the between-run CV was ≤ 7.38% for GDC-0834 and ≤ 8.20% for M1 metabolite. Between run percentage bias ranged from -2.76 to 6.98% for GDC-0834 and from -6.73 to 2.21% for M1 metabolite. GDC-0834 and M1 metabolite were stable in human plasma for 31 days at -20 and -70°C. This method was successfully applied to support a GDC-0834 human pharmacokinetic-based study.

  18. Development, validation and application of a 96-well enzymatic assay based on LC-ESI-MS/MS quantification for the screening of selective inhibitors against Mycobacterium tuberculosis purine nucleoside phosphorylase.

    PubMed

    Cattaneo, Giulia; Ubiali, Daniela; Calleri, Enrica; Rabuffetti, Marco; Höfner, Georg C; Wanner, Klaus T; De Moraes, Marcela C; Martinelli, Leonardo K B; Santos, Diógenes Santiago; Speranza, Giovanna; Massolini, Gabriella

    2016-11-02

    Mycobacterium tuberculosis (Mtb) purine nucleoside phosphorylase (PNP, EC 2.4.2.1) has been identified as a target for the development of specific inhibitors with potential antimycobacterial activity. We hereby described the development and validation of a new 96-well LC-ESI-MS/MS method to assess the inhibition activity of nucleoside analogues towards MtbPNP and the human PNP (HsPNP). Enzyme activity was determined by monitoring the phosphorolysis of inosine (Ino) to hypoxanthine (Hpx). The enzymatic assay (v = 0.5 mL, enzyme<0.2 μg/well, T = 37 °C) was performed with an overall time of about 15 min/plate for sample processing and 2 min/sample for LC-MS analysis. Validation of the quantification method met the criteria of the CDER guidance of FDA. Kinetic parameters were in agreement with those reported in literature (HsPNP KM = 0.150 ± 0.020 mM vs 0.133 ± 0.015 mM; MtbPNP KM = 0.060 ± 0.009 mM vs 0.040 ± 0.003 mM for Ino), thus demonstrating the reliability of the newly developed enzymatic assay. Preliminary inhibition assays confirmed the effects reported for Acyclovir (Acv) and Formycin A (FA) against HsPNP and MtbPNP. The validated enzymatic assay was applied to the evaluation of a set of 8-halo-, 8-amino-, 8-O-alkyl-substituted purine ribonucleosides synthesized on purpose as potential inhibitors against MtbPNP. The assayed 8-substituted ribonucleosides did not exert a significant inhibitory effect against the tested enzymes up to 1 mM.

  19. A direct heating model to overcome the edge effect in microplates.

    PubMed

    Lau, Chun Yat; Zahidi, Alifa Afiah Ahmad; Liew, Oi Wah; Ng, Tuck Wah

    2015-01-01

    Array-based tests in a microplate format are complicated by the regional variation in results of the outer against the inner wells of the plate. Analysis of the evaporation mechanics of sessile drops showed that evaporation rate increase with temperature was due to changes in the heat of vaporization, density and diffusion coefficient. In simulations of direct bottom heating of standard microplates, considerable heat transfer via conduction from the side walls was found to be responsible for lower temperatures in the liquid in wells close to the edge. Applying a two temperature heating mode, 304 K at the side compared to 310 K at the bottom, allowed for a more uniform temperature distribution. Transparency microplates were found to inherently possess immunity to the edge effect problem due to the presence of air between the liquid and solid wall.

  20. Rigid and non-rigid micro-plates: Philippines and Myanmar-Andaman case studies

    NASA Astrophysics Data System (ADS)

    Rangin, Claude

    2016-01-01

    Generally, tectonic plates are considered as rigid. Oblique plate convergence favors the development of micro-plates along the converging boundaries. The north-south-trending Philippines archipelago (here named Philippine Mobile Belt, PMB), a few hundreds kilometers wide, is one of such complex tectonic zones. We show here that it is composed of rigid rotating crustal blocks (here called platelets). In Myanmar, the northernmost tip of the Sumatra-Andaman subduction system is another complex zone made of various crustal blocks in-between convergent plates. Yet, contrary to PMB, it sustains internal deformation with platelet buckling, altogether indicative of a non-rigid behavior. Therefore, the two case studies, Philippine Mobile Belt and Myanmar-Andaman micro-plate (MAS), illustrate the complexity of micro-plate tectonics and kinematics at convergent plate boundaries.

  1. Events & Opinion: A Report to Our Readers.

    ERIC Educational Resources Information Center

    Green, Joseph H.

    1995-01-01

    Reprints an article originally published in 1957. Surveys readers of the journal. Notes that readers felt that 89% of the articles were either very good or excellent. Lists readers' views of pressing educational issues. (RS)

  2. Crustal Accretion and Mantle Geodynamics at Microplates: Constraints from Gravity Analysis

    NASA Astrophysics Data System (ADS)

    Ames, K.; Georgen, J. E.; Dordevic, M. M.

    2013-12-01

    Oceanic crustal accretion occurs in a variety of locations, including mid-ocean ridges and back-arc spreading centers, and in unique settings within these systems, such as plate boundary triple junctions, intra-transform spreading centers, and microplates. This study focuses on crustal accretion and mantle geodynamics at microplates. The Easter and Juan Fernandez microplates are located in the South Pacific along the Pacific, Nazca and Antarctic plate boundaries. Both microplates formed 3-5 Ma and they are currently rotating clockwise at 15 deg/Ma and 9 deg/Ma respectively (e.g., Searle et al. J. Geol. Soc. Lond. 1993). The study area also encompasses the Easter/Sala y Gomez mantle plume and the Foundation seamount chain, both of which are located close to spreading centers. We calculate mantle Bouguer anomaly (MBA) from satellite gravity measurements and shipboard soundings in order to gain a better understanding of the thermal structure of these two oceanic microplates and to quantify the effect that melting anomalies may have on their boundaries. We assume a crustal thickness of 6.0 km, a 1.7 g/cm^3 density difference at the water/crust interface, and a 0.6 g/cm^3 density difference at the crust/mantle interface. The west rift of the Easter microplate has an MBA low ranging from approximately -50 to -100 mGal, while the east rift has slightly higher MBA values ranging from roughly 10 to -50 mGal. The west rift of the Juan Fernandez microplate has a maximum MBA low of about -100 mGal with a sharp increase to -20 mGal at -35 deg S. The east rift of the Juan Fernandez microplate is characterized by more variable MBA, ranging from 0 to -140 mGal. The MBA low associated with the Easter/Sala y Gomez mantle plume has a maximum amplitude about 150 mGal. Likewise, the Foundation seamounts show a gravity low of -140 to -150 mGal. These spatial variations in gravity, as well as published isotopic data and exploratory numerical models, are used to constrain upper mantle

  3. Microplate-compatible total internal reflection fluorescence microscopy for receptor pharmacology

    NASA Astrophysics Data System (ADS)

    Chen, Minghan; Zaytseva, Natalya V.; Wu, Qi; Li, Min; Fang, Ye

    2013-05-01

    We report the use of total internal reflection fluorescence (TIRF) microscopy for analyzing receptor pharmacology and the development of a microplate-compatible TIRF imaging system. Using stably expressed green fluorescence protein tagged β2-adrenergic receptor as the reporter, we found that the activation of different receptors results in distinct kinetic signatures of the TIRF intensity of cells. These TIRF signatures closely resemble the characteristics of their respective label-free dynamic mass redistribution signals in the same cells. This suggests that TIRF in microplate can be used for profiling and screening drugs.

  4. Coupling the Torpedo microplate-receptor binding assay with mass spectrometry to detect cyclic imine neurotoxins.

    PubMed

    Aráoz, Rómulo; Ramos, Suzanne; Pelissier, Franck; Guérineau, Vincent; Benoit, Evelyne; Vilariño, Natalia; Botana, Luis M; Zakarian, Armen; Molgó, Jordi

    2012-12-04

    Cyclic imine neurotoxins constitute an emergent family of neurotoxins of dinoflagellate origin that are potent antagonists of nicotinic acetylcholine receptors. We developed a target-directed functional method based on the mechanism of action of competitive agonists/antagonists of nicotinic acetylcholine receptors for the detection of marine cyclic imine neurotoxins. The key step for method development was the immobilization of Torpedo electrocyte membranes rich in nicotinic acetylcholine receptors on the surface of microplate wells and the use of biotinylated-α-bungarotoxin as tracer. Cyclic imine neurotoxins competitively inhibit biotinylated-α-bungarotoxin binding to Torpedo-nicotinic acetylcholine receptors in a concentration-dependent manner. The microplate-receptor binding assay allowed rapid detection of nanomolar concentrations of cyclic imine neurotoxins directly in shellfish samples. Although highly sensitive and specific for the detection of neurotoxins targeting nicotinic acetylcholine receptors as a class, the receptor binding assay cannot identify a given analyte. To address the low selectivity of the microplate-receptor binding assay, the cyclic imine neurotoxins tightly bound to the coated Torpedo nicotinic receptor were eluted with methanol, and the chemical nature of the eluted ligands was identified by mass spectrometry. The immobilization of Torpedo electrocyte membranes on the surface of microplate wells proved to be a high-throughput format for the survey of neurotoxins targeting nicotinic acetylcholine receptors directly in shellfish matrixes with high sensitivity and reproducibility.

  5. Readers' Theatre as a History Teaching Tool.

    ERIC Educational Resources Information Center

    Harmon, Sandra D.; Riney-Kehrberg, Pamela; Westbury, Susan

    1999-01-01

    Describes the production of a readers' theater version of the first women's rights convention held at Seneca Falls, New York (includes script). Discusses the benefits of readers' theater for studying women's history and encouraging female student class participation; strategies for using readers' theater; and students creating their own readers'…

  6. Bringing Out the "Super Reader" in Everyone.

    ERIC Educational Resources Information Center

    Reichelderfer, Melissa A.

    2001-01-01

    Describes a "Super Reader" project that gives students an opportunity to show off their ability as readers. Defines "Super Readers" as students who read aloud to small groups of their peers. Notes that the "Super Reader" project encourages students to practice good reading behaviors, which are important for holding…

  7. Cognitive Profiles of Korean Poor Readers

    ERIC Educational Resources Information Center

    Cho, Jeung-Ryeul; Ji, Yu-Kyong

    2011-01-01

    This study compared the performance of 30 poor readers in the third grade with those of 30 average readers of the same age and 30 younger readers matched with the same reading level on phonological, visuo-perceptual, orthographic, and naming speed tasks. Individual data revealed heterogeneous profiles for the poor readers: six (20%) exhibited…

  8. Developing Lifelong Readers: Policies, Procedures, and Programs.

    ERIC Educational Resources Information Center

    Hayes, Bernard L., Ed.; Camperell, Kay, Ed.

    1992-01-01

    Selected papers from the 1991 annual conference of the American Reading Forum are presented in this yearbook. The papers, which deal with issues regarding ways to develop and strengthen a society of active readers, address policies, procedures, and programs for the beginning child reader, the beginning adult reader, and a range of readers in…

  9. Toward a Better Understanding of Reader Analysis.

    ERIC Educational Resources Information Center

    Suchan, James; Dulek, Ron

    1988-01-01

    Asserts that current business communication theory and practice make new inroads into reader analysis difficult. Discusses four pedagogical problems in reader analysis, including confusion over critical terminology, and poor heuristics for assessing readers. Explores readers' perceptual sets as a way of more accurately modeling the dynamics of the…

  10. New GPS Constrains on Crustal Deformation within the Puerto Rico-Virgin Islands Microplate

    NASA Astrophysics Data System (ADS)

    Solares, M. M.; Lopez, A. M.; Jansma, P. E.; Mattioli, G. S.

    2015-12-01

    Over twenty years of Global Positioning System (GPS) observations along the northeastern region of the Caribbean plate boundary zone have been used to evaluate crustal deformation in the Puerto Rico and Virgin Islands (PRVI) microplate, which generally translates westward relative to the Caribbean plate. New data from continuous GPS stations (cGPS) and re-occupied campaign GPS stations (eGPS) obtained between 2014 and 2015 allowed us to update the velocity field of the PRVI GPS Network and redefine the existing plate kinematics model of the PRVI microplate from previous measurements (Jansma et al., 2000; Jansma & Mattioli, 2005). Geodetic datasets for this epoch were processed with GIPSY/OASIS II (v.6.2) using an absolute point positioning strategy with final, precise orbits and clocks from JPL (IGS08). Results of sites velocity in the PRVI block are presented with respect to North America and Caribbean reference frames in ITRF08. The horizontal velocity components were used to calculate baseline lengths changes between selected GPS stations that span on-land faults and microplate boundaries, thus allowing quantification of internal deformation within the PRVI block. This enables us to locate zones of active deformation and faulting in order to understand how the relative motion between geological structures is accommodated. Our updated velocity field constrains intraplate deformation to 1-3 mm/yr across the PRVI microplate and active extension of 1-2 mm/yr in the Anegada passage eastern boundary. In addition, counterclockwise rotation has been observed and may be related to the deformation in southwestern Puerto Rico continuing offshore to the Muertos Trough along PRVI's southern boundary. Despite the PRVI microplate slow motion and small deformation, increasing velocities from east to west coincides with the most active microseismic zone and ongoing deformation in southwestern Puerto Rico suggesting independent motion along this segment of the PRVI block.

  11. Enhancement of Enzymatic Colorimetric Response by Silver Island Films on High Throughput Screening Microplates

    PubMed Central

    Abel, Biebele; Clement, Travis C.; Aslan, Kadir

    2014-01-01

    In this study, we report the use of an enzyme-based hybrid platform, which is comprised of silver island films, enzymes (HRP and AP) and high-throughput screening (HTS) microplates, to enhance the colorimetric response of enzymatic reactions. The hybrid platform was designed in a two-step process: (i) deposition of SIFs onto HTS microplates with low, medium, and high loading (refers to the extent of the surface plasmon resonance peak of SIFs at 460 nm) using Tollen’s reaction scheme; and (ii) attachment of b-BSA or BEA as linkers for the immobilization of enzymes. The presence of SIFs within the wells of the HTS microplates was confirmed using an optical spectrophotometer and real-color photography. Control experiments, where SIFs were omitted from the surfaces were carried out to confirm the effect of SIFs on the enzymatic colorimetric response. Significant colorimetric signal enhancement was observed for HRP or AP on SIFs (high loading) deposited HTS microplates using b-BSA (up to ~ 3-fold for AP and ~6-fold HRP) or BEA (up to ~ 7-fold for both HRP and AP), as compared to our control samples. The observed increase in colorimetric response can be attributed to the nature of BEA, which exposes surface-bound enzymes to the substrate present in bulk more efficiently than b-BSA. This study proves that SIFs can serve as a valuable tool to improve the signal output of existing bioassays carried out in HTS microplates, which can be applicable to the field biosensors and plasmonics. PMID:24950456

  12. Narratives of the Struggling Reader

    ERIC Educational Resources Information Center

    Vlach, Saba; Burcie, Judy

    2010-01-01

    Struggling readers need to be taught at their instruction level. But often, separating them from their classmates reinforces their feelings of inadequacy. This article will explain the importance of developing a sense of agency in struggling learners and outline some strategies teachers can incorporate to help make this happen within the classroom.

  13. Fairy Tales for Two Readers.

    ERIC Educational Resources Information Center

    Criscoe, Betty L., Ed.; Lanasa, Philip J., III, Ed.

    The 15 adapted fairy tales presented in this book were prepared for use in practicing oral reading by a parent and a child, a teacher and a child, or two children, one of whom reads slightly better than the other. The stories in the book are arranged in dialogue format for two readers. The high interest/low readability stories in the book are…

  14. Becoming a Classroom of Readers

    ERIC Educational Resources Information Center

    Miller, Donalyn

    2010-01-01

    Studies have shown that voracious reading improves students' academic performance in a variety of realms. In this article, 6th grade teacher Donalyn Miller explains how she has helped her students become committed readers. Her students are required to read at least 40 books each year in a variety of genres and styles. She gives them time to read…

  15. Reader Response in the Classroom.

    ERIC Educational Resources Information Center

    Chew, Charles, Ed.; And Others

    Focusing on reader response in the classroom, the works collected in this book represent the results of a five-week summer institute in which 25 middle school, high school, and college teachers studied the principles and applications of literature instruction. The following essays are included: an introduction by G. Garber; "An Overview of the…

  16. Readers' Emotions: A Plurilingual Perspective

    ERIC Educational Resources Information Center

    Rivière, Marie

    2016-01-01

    This article focuses on the differential emotional power of languages in the book-reading practices of plurilingual readers. Within a plurilingual perspective, it aims at adding nuance to the "emotional contexts of learning hypothesis" and the "theory of language embodiment". This qualitative study is based on semi-structured…

  17. Roots: An Asian American Reader.

    ERIC Educational Resources Information Center

    Tachiki, Amy, Ed.; And Others

    A documentary collection of the experiences of Asian Americans from a multitude of perspectives, including a scholarly focus and also containing contemporary expressions, comprises "Roots: An Asian American Reader." The volume is said to be designed to meet the needs of Asian Americans by providing a compilation of materials in readily…

  18. Readers' Knowledge of Popular Genre

    ERIC Educational Resources Information Center

    Dixon, Peter; Bortolussi, Marisa

    2009-01-01

    This research examined readers' knowledge of popular genres. Participants wrote short essays on fantasy, science fiction, or romance. The similarities among the essays were measured using latent semantic analysis (LSA) and were then analyzed using multidimensional scaling and cluster analysis. The clusters and scales were interpreted by searching…

  19. Building a Nation of Readers.

    ERIC Educational Resources Information Center

    Library of Congress, Washington, DC. Center for the Book.

    Inviting individuals and families, schools, libraries, educational, civic and government associations, labor unions, and others to join in building a nation of readers, this booklet lists ideas for organizing and supporting reading promotion projects that benefit all age groups. Ideas presented in the booklet include: teach someone to read; set…

  20. Comparative Education. ASHE Reader Series.

    ERIC Educational Resources Information Center

    Kempner, Ken, Ed.; Mollis, Marcela, Ed.; Tierney, William G., Ed.

    The chapters in this collection explore why particular national higher education systems operate as they do and the effects these systems have on one another, on national and global development, and on the production of knowledge. The works included in this reader address the assessment of inputs and outputs of institutions and the meaning these…

  1. Serving Boys through Readers' Advisory

    ERIC Educational Resources Information Center

    Sullivan, Michael

    2010-01-01

    Based on more than twenty years' experience working to get boys interested in reading, the author now offers his first readers' advisory volume. With an emphasis on nonfiction and the boy-friendly categories of genre fiction, the work offers a wealth of material including: (1) Suggestions for how to booktalk one-on-one as well as in large groups;…

  2. Optical Calibration of TLD Readers

    DTIC Science & Technology

    2007-11-02

    DISTRIBUTION CODE 13. ABSTRACT: The Navy uses thermoluminescent dosimeters (TLDs) on all of its nuclear warships. TLDs measure the amount of radiation a person...6614. SUBJECT TERMS: Nucl_______ thermoluminescent dosimeters, TLD reader, temperature compensation, laser diodes, light emitting diodes...Navy uses thermoluminescent dosimeters (TLDs) on all of its nuclear warships. TLDs measure the amount of radiation a person receives through the use

  3. Books for Adult New Readers.

    ERIC Educational Resources Information Center

    O'Brien, Roberta Luther, Comp.

    This document is an annotated bibliography of recommended print materials for English-speaking adults reading at the seventh grade level or below. (Sixty percent of the titles are at fifth grade level or below). The titles were selected for their broad appeal to the average adult new reader. In the selection, special consideration was given to…

  4. Writing Reviews for Readers' Advisory

    ERIC Educational Resources Information Center

    Hooper, Brad

    2010-01-01

    Reviews are an important resource for readers' advisory and collection development. They are also a helpful promotional tool, introducing patrons to what is new on the shelf. This resource includes: (1) Tips for writing strong, relevant reviews; (2) Different ways reviews can be used to promote your library; and (3) A chapter by Joyce Saricks…

  5. Spectral Unmixing Plate Reader: High-Throughput, High-Precision FRET Assays in Living Cells.

    PubMed

    Schaaf, Tory M; Peterson, Kurt C; Grant, Benjamin D; Thomas, David D; Gillispie, Gregory D

    2017-03-01

    We have developed a microplate reader that records a complete high-quality fluorescence emission spectrum on a well-by-well basis under true high-throughput screening (HTS) conditions. The read time for an entire 384-well plate is less than 3 min. This instrument is particularly well suited for assays based on fluorescence resonance energy transfer (FRET). Intramolecular protein biosensors with genetically encoded green fluorescent protein (GFP) donor and red fluorescent protein (RFP) acceptor tags at positions sensitive to structural changes were stably expressed and studied in living HEK cells. Accurate quantitation of FRET was achieved by decomposing each observed spectrum into a linear combination of four component (basis) spectra (GFP emission, RFP emission, water Raman, and cell autofluorescence). Excitation and detection are both conducted from the top, allowing for thermoelectric control of the sample temperature from below. This spectral unmixing plate reader (SUPR) delivers an unprecedented combination of speed, precision, and accuracy for studying ensemble-averaged FRET in living cells. It complements our previously reported fluorescence lifetime plate reader, which offers the feature of resolving multiple FRET populations within the ensemble. The combination of these two direct waveform-recording technologies greatly enhances the precision and information content for HTS in drug discovery.

  6. Sulfurizing-Induced Hollowing of Co9S8 Microplates with Nanosheet Units for Highly Efficient Water Oxidation.

    PubMed

    Liu, Huan; Ma, Fei-Xiang; Xu, Cheng-Yan; Yang, Li; Du, Yue; Wang, Pan-Pan; Yang, Shuang; Zhen, Liang

    2017-03-14

    Transition metal-based compounds are promising alternative non-precious electrocatalysts for hydrogen and oxygen evolution to noble metals-based materials. Nanosheets-constructed hollow structures can efficiently promote the electrocatalystic activity, mainly because of their largely exposed active sites. Herein, hierarchical Co9S8 hollow microplates with nanosheet building units are fabricated via sulfurization and subsequent calcination of pre-formed Co-glycolate microplates. Benefited from the advantages of hollow structure, nanosheet units and high Co3+ content, Co9S8 hollow microplates exhibit remarkable catalytic performance for oxygen evolution reaction (OER) with low overpotential of 278 mV to reach current density of 10 mA cm-2, low Tafel slope of 53 mV dec-1 and satisfied stability. This construction method of Co9S8 hierarchical hollow microplates composing by nanosheets structure is an effective tactics for promoting OER performance of water splitting electrocatalysts.

  7. Seismic evidence for rotating mantle flow around subducting slab edge associated with oceanic microplate capture

    NASA Astrophysics Data System (ADS)

    Mosher, Stephen G.; Audet, Pascal; L'Heureux, Ivan

    2014-07-01

    Tectonic plate reorganization at a subduction zone edge is a fundamental process that controls oceanic plate fragmentation and capture. However, the various factors responsible for these processes remain elusive. We characterize seismic anisotropy of the upper mantle in the Explorer region at the northern limit of the Cascadia subduction zone from teleseismic shear wave splitting measurements. Our results show that the mantle flow field beneath the Explorer slab is rotating anticlockwise from the convergence-parallel motion between the Juan de Fuca and the North America plates, re-aligning itself with the transcurrent motion between the Pacific and North America plates. We propose that oceanic microplate fragmentation is driven by slab stretching, thus reorganizing the mantle flow around the slab edge and further contributing to slab weakening and increase in buoyancy, eventually leading to cessation of subduction and microplate capture.

  8. Characterization of protein-membrane binding interactions via a microplate assay employing whole liposome immobilization.

    PubMed

    Smith, Matthew D; Best, Michael D

    2011-01-01

    Protein-cell membrane binding interactions control numerous vital biological processes, many of which can go awry during disease onset. However, the study of these events is complicated by the complexity of the membrane bilayer. These efforts would benefit from a rapid and easily accessible method for characterizing protein-membrane recognition events. Herein, we describe a microplate-based method for the detection of protein-membrane binding that employs whole liposome immobilization using a biotin anchor. First, control studies are detailed to test for nonspecific liposome immobilization (fluorescence assay; see Subheading 3.2), and to ensure that liposomes remain intact on the microplate surface (dye leakage assay; see Subheading 3.3). Finally, a protein-membrane binding detection assay is described through the example of protein kinase Cα binding to surface-immobilized whole liposomes (see Subheading 3.4).

  9. A Fast Optical Method for the Determination of Liquid Levels in Microplates

    PubMed Central

    Thurow, Kerstin; Stoll, Norbert; Ritterbusch, Kai

    2011-01-01

    Parallel liquid handling systems are widely used in different applications of life sciences. In order to avoid false positive or negative results which lead to higher costs due to the replication of the experiments it is necessary to monitor the process and success of liquid delivery. An easy method for the determination of the liquid levels in microplates has been developed and evaluated. The optical method bases on the measurement of the liquid level using CCD cameras followed by special algorithms for the evaluation and visualization of the measured data. The proposed method was tested in changing environmental lighting for two different liquids. These tests confirm our approach towards optical liquid level determination for smallest volumes in microplates and also show the challenges regarding environmental lighting and different physical properties of fluids. PMID:21747734

  10. Quantitative, competitive PCR assay for HIV-1 using a microplate-based detection system.

    PubMed

    Guenthner, P C; Hart, C E

    1998-05-01

    We have developed a quantitative competitive PCR (QC-PCR) assay in a microplate format for quantifying human immunodeficiency virus Type 1 (HIV-1) DNA or RNA in a broad range of source materials. Our QC-PCR assay is a modification of technique originally described by Piatak et al. (1993), which is based on the presence of a competitive internal standard containing an internal 80-bp deletion of HIV-1 gag target sequence. For improved detection and quantification of the wild-type and internal-standard PCR products in a microplate format, we introduced a non-HIV, 31-bp insert into the internal standard as a probe hybridization site that does not cross-hybridize with wild-type HIV-1 products. By using a primer pair in which one primer is biotinylated, QC-PCRs can be bound to a streptavidin-coated microplate, denatured and probed with a digoxigenin (Dig)-labeled, wild-type or internal-standard probe. The hybridized Dig-labeled probes are detected with an anti-Dig antibody conjugated to detector molecules for luminometry (aequorin) or optical densitometry (peroxidase), yielding results that are quantifiable over the range of 100-10,000 copies of HIV gag. Tested source materials for HIV-1 DNA or RNA quantification include plasma, vaginal lavage and cultured cells. The application of the QC-PCR assay using the microplate format affords a convenient and cost-effective method for quantifying HIV-1 proviral and viral loads from a variety of body fluids, cells and tissues.

  11. Detection of possible economically motivated adulterants in heparin sodium and low molecular weight heparins with a colorimetric microplate based assay.

    PubMed

    Sommers, Cynthia D; Keire, David A

    2011-09-15

    Recently, we described a 96-well plate format assay for visual detection of oversulfated chondroitin sulfate A (OSCS) contamination in heparin samples based on a water-soluble cationic polythiophene polymer (3-(2-(N-(N'-methylimidazole))ethoxy)-4-methylthiophene (LPTP)) and heparinase digestion of heparin. Here, we establish the specificity of the LPTP/heparinase test with a unique set of reagents that define the structural requirements for significant LPTP chemosensor color change. For example, we observed a biphasic behavior of larger shifts to the red in the UV absorbance spectra with decreasing average molecular weight of heparin chains with a break below 12-mer chain lengths. In addition, the oversulfation of chondroitin sulfate A (CSA) to a partially (PSCS) or fully (OSCS) sulfated form caused progressively less red shift of LPTP solutions. Furthermore, glycosaminoglycans (GAGs) containing glucuronic acid caused distinct spectral patterns compared to iduronic acid containing GAGs. We applied the LPTP/heparinase test to detection of OSCS (≥0.03% (w/w) visually or 0.01% using a plate reader) in 10 μg amounts of low molecular weight heparins (LMWHs; i.e. dalteparin, tinzaparin, or enoxaparin). Furthermore, because other oversulfated GAGs are possible economically motivated adulterants (EMAs) in heparin sodium, we tested the capacity of the LPTP/heparinase assay to detect oversulfated dermatan sulfate (OSDS), heparin (OSH), and heparan sulfate (OSHS). These potential EMAs were visually detectable at a level of ∼0.1% when spiked into heparin sodium. We conclude that the LPTP/heparinase test visually detects oversulfated GAGs in heparin sodium and LMWHs in a format potentially amenable to high-throughput screening.

  12. High-throughput microplate enzymatic assays for fast sugar and acid quantification in apple and tomato.

    PubMed

    Vermeir, S; Nicolaï, B M; Jans, K; Maes, G; Lammertyn, J

    2007-05-02

    In this article, we report on the use of miniaturized and automated enzymatic assays as an alternative technology for fast sugar and acid quantification in apples and tomatoes. Enzymatic assays for d-glucose, d-fructose, sucrose, D-sorbitol/xylitol, L-malic acid, citric acid, succinic acid, and L-glutamic acid were miniaturized from the standard 3 mL assays in cuvettes into assays of 200 microL or lower in 96 or 384 well microplates. The miniaturization and the automation were achieved with a four channel automatic liquid handling system in order to reduce the dispensing errors and to obtain an increased sample throughput. Performance factors (limit of detection, linearity of calibration curve, and repeatability) of the assays with standard solutions were proven to be satisfactory. The automated and miniaturized assays were validated with high-pressure liquid chromatography (HPLC) analyses for the quantification of sugars and acids in tomato and apple extracts. The high correlation between the two techniques for the different components indicates that the high-throughput microplate enzymatic assays can serve as a fast, reliable, and inexpensive alternative for HPLC as the standard analysis technique in the taste characterization of fruit and vegetables. In addition to the analysis of extracts, the high-throughput microplate enzymatic assays were used for the direct analysis of centrifuged and filtered tomato juice with an additional advantage that the sample preparation time and analysis costs are reduced significantly.

  13. Application of a microplate scale fluorochrome staining assay for the assessment of viability of probiotic preparations.

    PubMed

    Alakomi, H-L; Mättö, J; Virkajärvi, I; Saarela, M

    2005-07-01

    Cell viability in probiotic preparations is traditionally assessed by the plate count technique. Additionally, fluorescent staining combined with epifluorescence microscopy or flow cytometry has been developed for the viability assessment, but the currently available assays are either laborious or require highly sophisticated equipment. The aim of this study was to investigate the applicability of a microplate scale fluorochrome assay for predicting the cell state of freeze-dried Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis preparations. In addition to viability assessment with LIVE/DEAD BacLight Bacterial Viability Kit, DiBAC(4)3 stain was used for the kinetic measurement of changes in bifidobacterial cell membrane functions during exposure to low pH. The microplate scale fluorochrome assay results on the viability and cell numbers of probiotic preparations correlated well with the results obtained with the culture-based technique and (with few exceptions) with epifluorescence microscopy. The assay was applicable also for the viability assessment of stressed (acid-treated) cells provided that the cell density in treatments was adjusted to the optimal measurement level of the fluorometer. The microplate scale fluorochrome assay offers a rapid and robust tool for the viability assessment of probiotic preparations, and enables also kinetic measurements.

  14. Wafer-scale growth of large arrays of perovskite microplate crystals for functional electronics and optoelectronics

    PubMed Central

    Wang, Gongming; Li, Dehui; Cheng, Hung-Chieh; Li, Yongjia; Chen, Chih-Yen; Yin, Anxiang; Zhao, Zipeng; Lin, Zhaoyang; Wu, Hao; He, Qiyuan; Ding, Mengning; Liu, Yuan; Huang, Yu; Duan, Xiangfeng

    2015-01-01

    Methylammonium lead iodide perovskite has attracted intensive interest for its diverse optoelectronic applications. However, most studies to date have been limited to bulk thin films that are difficult to implement for integrated device arrays because of their incompatibility with typical lithography processes. We report the first patterned growth of regular arrays of perovskite microplate crystals for functional electronics and optoelectronics. We show that large arrays of lead iodide microplates can be grown from an aqueous solution through a seeded growth process and can be further intercalated with methylammonium iodide to produce perovskite crystals. Structural and optical characterizations demonstrate that the resulting materials display excellent crystalline quality and optical properties. We further show that perovskite crystals can be selectively grown on prepatterned electrode arrays to create independently addressable photodetector arrays and functional field effect transistors. The ability to grow perovskite microplates and to precisely place them at specific locations offers a new material platform for the fundamental investigation of the electronic and optical properties of perovskite materials and opens a pathway for integrated electronic and optoelectronic systems. PMID:26601297

  15. Using Readers' Theatre in the Classroom.

    ERIC Educational Resources Information Center

    Annarella, Lorie A.

    Reader's Theatre can be used to combine basic literature and writing instruction with creative arts. Improvisational playmaking by students, using literature in the form of plays, prose, and poetry, forms the basis of Reader's Theatre. Use of Reader's Theatre in the classroom can: (1) foster deeper understanding of character, setting, and plot…

  16. Benefit of the Doubt. Reader Reflections

    ERIC Educational Resources Information Center

    Mathematics Teacher, 2016

    2016-01-01

    The editors of Mathematics Teacher appreciate the interest of readers and value the views of those who write in with comments. The editors ask that name and affiliation including email address be provided at the end of their letters. This September 2016 Reader Reflections, provides reader comments on the following articles: (1) "Innocent…

  17. E-Readers: Powering up for Engagement

    ERIC Educational Resources Information Center

    Miranda, Twyla; Johnson, Kary A.; Rossi-Williams, Dara

    2012-01-01

    E-readers like the Amazon Kindle and the Barnes and Noble Nook are beginning to make their way into school libraries and classrooms. It's about time. E-readers have tremendous potential to entice reluctant readers to read more. A study that the authors recently conducted among low-reading-ability middle school students demonstrated that potential.…

  18. Block rotation and continental extension in Afar: A comparison to oceanic microplate systems

    NASA Astrophysics Data System (ADS)

    Acton, Gary D.; Stein, Seth; Engeln, Joseph F.

    1991-06-01

    The reorganization of oceanic spreading centers separating major plates often appears to occur by a process in which discrete microplates form and evolve by rift propagation. To see whether such microplate behavior has implications for continental rifting, we investigate the application of a microplate model to the Afar region at the Nubia-Somalia-Arabia triple junction. Studies of marine magnetic anomalies, volcanic ages, bathymetry, and seismicity suggest that the westward propagating Gulf of Aden spreading center has propagated into eastern Afar within the past 2 m.y., causing rifting and extension within the continent. We derive constraints on the extension history from the geometry and timing of rift formation and from paleomagnetic data indicating that Pliocene to Pleistocene age rocks have undergone a clockwise rotation of ˜11°. We suggest that the history of rifting, the rotation, and several other features of the regional geology can be described by combining features of an oceanic microplate model and the concept of rift localization previously proposed for Afar. In this scenario, motion occurring on several rifts within an extensional zone preceding the propagating spreading center is gradually transferred to a single rift. While motion is transferred, the overlap region between the growing and dying rifts acts as one or more microplates or blocks that rotate relative to the surrounding major plates. The rifting history and rotations in eastern Afar are thus related to the rift propagation and localization that occurs as the plate boundary evolves. Provided the constraints we use are appropriate, our model better describes the regional kinematics than alternative block models including one based on "bookshelf" faulting. If the tectonics of Afar are typical for continental breakup, they have interesting implications for the geometry of passive margins. In particular, asymmetric rifted margins can be produced if the final location of the rift axis is not

  19. Cerebral Dominance in Disabled Readers, Good Readers, and Gifted Children: Search for a Valid Model

    ERIC Educational Resources Information Center

    Kershner, John R.

    1977-01-01

    Disabled readers, good readers, and gifted children were compared on their recognition of words presented tachistoscopically under stimultaneous, bilateral viewing conditions. Poor readers showed inferior right-field performance compared with gifted and good readers, but when IQ was covaried, they showed higher left-field scores and lower visual…

  20. Variable Distance Angular Symbology Reader

    NASA Technical Reports Server (NTRS)

    Schramm, Harry F., Jr. (Inventor); Corder, Eric L. (Inventor)

    2006-01-01

    A variable distance angular symbology, reader utilizes at least one light source to direct light through a beam splitter and onto a target. A target may be angled relative to the impinging light beam up to and maybe even greater than 45deg. A reflected beam from the target passes through the beam splitter and is preferably directed 90deg relative to the light source through a telecentric lens to a scanner which records an image of the target such as a direct part marking code.

  1. Faulting and strain partitioning in Jamaica from GPS and structural data: Implications for Gonave and Hispaniola microplate kinematics, northern Caribbean

    NASA Astrophysics Data System (ADS)

    Benford, Bryn

    A series of small microplates separate the Caribbean and North America plates in the northern Caribbean. My dissertation focuses on understanding the structural evolution and neotectonic deformation of Jamaica, and how it relates to the overall microplates and tectonics of the northern Caribbean. Jamaica, which lies along the same seismically active plate boundary as Haiti, has had twelve earthquakes with Modified Mercalli intensities of VII to X since 1667. However, remarkably little is known about which faults presently constitute the most significant seismic hazards. This research provides insight into tectonic processes and facilitates mitigation of geological hazards in the region. Two chapters focus on characterizing deformation in Jamaica through modeling GPS velocities and through field mapping. The best-fitting models based on GPS velocities place most strike-slip motion on faults in central Jamaica and suggest that faults in northern Jamaica have minimal motion. I estimate 4--5 mm yr-1 of slip for faults near the capital city of Kingston of southeastern Jamaica, implying significant seismic hazard. Field mapping combined with present-day topography, focal mechanisms, geology, gravity, and well and borehole data indicate that east-west contraction is accommodated by reactivated, NNW-striking reverse faults, which are bound by E-striking strike-slip faults in southern Jamaica. The other two chapters of my thesis focuses on understanding the behavior of the microplates along the Caribbean-North America plate boundary: I model GPS velocities and use shear-wave splitting to understand the crustal and mantle behaviors, respectively of the microplates. The GPS data require an independently moving Hispaniola microplate between the Mona Passage and a likely diffuse boundary just west of or within western Hispaniola. The new microplate angular velocities predict 6.8+/-1.0 mm yr-1 of left-lateral slip and 5.7+/-1 mm yr-1 of convergent motion surrounding the

  2. Conventional tube and microplate Limulus amoebocyte lysate procedures for determination of gram-negative bacteria in milk.

    PubMed

    May, S A; Mikolajcik, E M; Richter, E R

    1989-05-01

    A comparison was made of the conventional tube and microplate Limulus amoebocyte lysate assay for detection of gram-negative bacterial lipopolysaccharide in milk. Raw whole milk samples were maintained frozen and portions were examined periodically on alternate days during 13-d storage to evaluate the reproducibility of both Limulus amoebocyte lysate procedures and to determine optimum reaction conditions for the microplate method. One-day-old, raw and locally purchased pasteurized milk samples, held at 7 degrees C, were analyzed during storage to establish the correlation of both procedures with aerobic and modified psychrotrophic plate counts. Vitamin- and mineral-fortified dairy-based products were examined using the microplate Limulus amoebocyte lysate test as a potential indicator of raw material or finished product bacterial quality and possible postprocessing contamination. Statistical analysis of the data collected comparing the conventional tube and the microplate Limulus amoebocyte lysate assay demonstrated no significant difference exists between the methods when either the modified psychrotrophic bacterial count or the aerobic plate count was used to determine gram-negative bacteria in pasteurized or raw milk (P less than .91). The microplate method, which uses half the lysate reagent, was a good indicator of the bacterial quality of milk and fortified dairy products, consistently detecting bacterial levels greater than 10(3) to 10(4)/ml.

  3. Microplate-based high throughput screening procedure for the isolation of lipid-rich marine microalgae

    PubMed Central

    2011-01-01

    We describe a new selection method based on BODIPY (4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene) staining, fluorescence activated cell sorting (FACS) and microplate-based isolation of lipid-rich microalgae from an environmental sample. Our results show that direct sorting onto solid medium upon FACS can save about 3 weeks during the scale-up process as compared with the growth of the same cultures in liquid medium. This approach enabled us to isolate a biodiverse collection of several axenic and unialgal cultures of different phyla. PMID:22192119

  4. Sensitive microplate assay for the detection of proteolytic enzymes using radiolabeled gelatin

    SciTech Connect

    Robertson, B.D.; Kwan-Lim, G.E.; Maizels, R.M.

    1988-07-01

    A sensitive, microplate assay is described for the detection of a wide range of proteolytic enzymes, using radio-iodine-labeled gelatin as substrate. The technique uses the Bolton-Hunter reagent to label the substrate, which is then coated onto the wells of polyvinyl chloride microtiter plates. By measuring the radioactivity released the assay is able to detect elastase, trypsin, and collagenase in concentrations of 1 ng/ml or less, while the microtiter format permits multiple sample handling and minimizes sample volumes required for analysis.

  5. A Sign Language Screen Reader for Deaf

    NASA Astrophysics Data System (ADS)

    El Ghoul, Oussama; Jemni, Mohamed

    Screen reader technology has appeared first to allow blind and people with reading difficulties to use computer and to access to the digital information. Until now, this technology is exploited mainly to help blind community. During our work with deaf people, we noticed that a screen reader can facilitate the manipulation of computers and the reading of textual information. In this paper, we propose a novel screen reader dedicated to deaf. The output of the reader is a visual translation of the text to sign language. The screen reader is composed by two essential modules: the first one is designed to capture the activities of users (mouse and keyboard events). For this purpose, we adopted Microsoft MSAA application programming interfaces. The second module, which is in classical screen readers a text to speech engine (TTS), is replaced by a novel text to sign (TTSign) engine. This module converts text into sign language animation based on avatar technology.

  6. [Readers' position against induced abortion].

    PubMed

    1981-08-25

    Replies to the request by the Journal of Nursing on readers' positions against induced abortion indicate there is a definite personal position against induced abortion and the assistance in this procedure. Some writers expressed an emotional "no" against induced abortion. Many quoted arguments from the literature, such as a medical dictionary definition as "a premeditated criminally induced abortion." The largest group of writers quoted from the Bible, the tenor always being: "God made man, he made us with his hands; we have no right to make the decision." People with other philosophies also objected. Theosophical viewpoint considers reincarnation and the law of cause and effect (karma). This philosophy holds that induced abortion impedes the appearance of a reincarnated being. The fundamental question in the abortion problem is, "can the fetus be considered a human life?" The German anatomist Professor E. Bleckschmidt points out that from conception there is human life, hence the fertilized cell can only develop into a human being and is not merely a piece of tissue. Professional nursing interpretation is that nursing action directed towards killing of a human being (unborn child) is against the nature and the essence of the nursing profession. A different opinion states that a nurse cares for patients who have decided for the operation. The nurse doesn't judge but respects the individual's decision. Some proabortion viewpoints considered the endangering of the mother's life by the unborn child, and the case of rape. With the arguments against abortion the question arises how to help the woman with unwanted pregnancy. Psychological counseling is emphasized as well as responsible and careful assistance. Referral to the Society for Protection of the Unborn Child (VBOK) is considered as well as other agencies. Further reader comments on this subject are solicited.

  7. Reader Response Theory and Classroom Practices.

    ERIC Educational Resources Information Center

    Bush, Harold K., Jr.

    1994-01-01

    Presents annotations of nine journal articles, monographs, and "learning packages" (published between 1989 and 1993) that examine teaching approaches founded upon the insights of reader response theory. (RS)

  8. Parallel electromembrane extraction in the 96-well format.

    PubMed

    Eibak, Lars Erik Eng; Rasmussen, Knut Einar; Oiestad, Elisabeth Leere; Pedersen-Bjergaard, Stig; Gjelstad, Astrid

    2014-05-30

    The repeatability and extraction recoveries of parallel electromembrane extraction (Pa-EME) was thoroughly investigated in the present project. Amitriptyline, fluoxetine, and haloperidol were isolated from eight samples of pure water, undiluted human plasma, and undiluted human urine, respectively; in total 24 samples were processed in parallel. The repeatability was found to be independent of the different sample matrices (pure water samples, human plasma, and water) processed in parallel, although the respective samples contained different matrix components. In another experiment seven of the 24 wells were perforated. Even though the perforation caused the total current level in the Pa-EME setup to increase, the intact circuits were unaffected by the collapse in seven of the circuits. In another approach, exhaustive extraction of amitriptyline, fluoxetine, and haloperidol was demonstrated from pure water samples. Amitriptyline and haloperidol were also isolated exhaustively from undiluted human plasma samples; the extraction recovery of fluoxetine from undiluted human plasma was 81%. Finally, the sample throughput was increased with the Pa-EME configuration. The extraction recoveries were investigated by processing 1, 8, 68, or 96 samples in parallel in 10min; neither the extraction recoveries nor the repeatability was affected by the total numbers of samples. Eventually, the Pa-EME was combined with ultra performance liquid chromatography (UPLC) to combine high-throughput sample preparation with high-throughput analytical instrumentation. The aim of the present investigation was to demonstrate the potential of electromembrane extraction as a high throughput sample preparation platform; and hopefully to increase the interest for EME in the bioanalytical field to solve exisiting and novel analytical challenges.

  9. Revision of the genera Microplitis and Snellenius (Hymenoptera, Braconidae, Microgastrinae) from Area de Conservacion Guanacaste, Costa Rica, with a key to all species previously described from Mesoamerica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genera Microplitis and Snellenius (Hymenoptera: Braconidae, Microgastrinae) from Area de Conservacion Guanacaste (ACG), Costa Rica, are revised. A total of 28 new species are described: 23 of Snellenius (the first record for Mesoamerica) and five of Microplitis. A key is provided to all new spec...

  10. High School Readers: A Profile of above Average Readers and Readers with Learning Disabilities Reading Expository Text

    ERIC Educational Resources Information Center

    Wigent, Catherine A.

    2013-01-01

    This study examined above average high school readers and high school readers with learning disabilities in order to better understand the impact of twelve years of formal education on reading skills and strategy use while reading expository text. This study examined reading strategies related to knowledge construction, monitoring, and evaluating…

  11. Metabolic activities of five botryticides against Botrytis cinerea examined using the Biolog FF MicroPlate

    PubMed Central

    Wang, Hancheng; Wang, Jin; Li, Licui; Hsiang, Tom; Wang, Maosheng; Shang, Shenghua; Yu, Zhihe

    2016-01-01

    Tobacco grey mold caused by Botrytis cinerea is an important fungal disease worldwide. Boscalid, carbendazim, iprodione, pyrimethanil and propiconazole are representative botryticides for grey mold management. This research investigated the sensitivities of B. cinerea from tobacco to these chemicals using the Biolog FF Microplate. All five chemicals showed inhibitory activity, with average EC50 values of 0.94, 0.05, 0.50, 0.61 and 0.31 μg ml−1, respectively. B. cinerea metabolized 96.8% of tested carbon sources, including 29 effectively and 33 moderately, but the metabolic fingerprints differed under pressures imposed by these botryticides. For boscalid, B. cinerea was unable to metabolize many substrates related to tricarboxylic acid cycle. For carbendazim, carbon sources related to glycolysis were not metabolized. For iprodione, use of most carbon substrates was weakly inhibited, and the metabolic profile was similar to that of the control. For propiconazole, no carbon substrates were metabolized and the physiological and biochemical functions of the pathogen were totally inhibited. These findings provide useful information on metabolic activities of these botryticides, and may lead to future applications of the Biolog FF Microplate for examining metabolic effects of other fungicides on other fungi, as well as providing a metabolic fingerprint of B. cinerea that could be useful for identification. PMID:27491536

  12. Size-dependent phase transition in methylammonium lead iodide perovskite microplate crystals

    SciTech Connect

    Li, Dehui; Wang, Gongming; Cheng, Hung -Chieh; Chen, Chih -Yen; Wu, Hao; Liu, Yuan; Huang, Yu; Duan, Xiangfeng

    2016-04-21

    Methylammonium lead iodide perovskite has attracted considerable recent interest for solution processable solar cells and other optoelectronic applications. The orthorhombic-to-tetragonal phase transition in perovskite can significantly alter its optical, electrical properties and impact the corresponding applications. Here, we report a systematic investigation of the size-dependent orthorhombic-to-tetragonal phase transition using a combined temperature-dependent optical, electrical transport and transmission electron microscopy study. Our studies of individual perovskite microplates with variable thicknesses demonstrate that the phase transition temperature decreases with reducing microplate thickness. The sudden decrease of mobility around phase transition temperature and the presence of hysteresis loops in the temperature-dependent mobility confirm that the orthorhombic-to-tetragonal phase transition is a first-order phase transition. Lastly, our findings offer significant fundamental insight on the temperature-and size-dependent structural, optical and charge transport properties of perovskite materials, and can greatly impact future exploration of novel electronic and optoelectronic devices from these materials.

  13. Size-dependent phase transition in methylammonium lead iodide perovskite microplate crystals

    DOE PAGES

    Li, Dehui; Wang, Gongming; Cheng, Hung -Chieh; ...

    2016-04-21

    Methylammonium lead iodide perovskite has attracted considerable recent interest for solution processable solar cells and other optoelectronic applications. The orthorhombic-to-tetragonal phase transition in perovskite can significantly alter its optical, electrical properties and impact the corresponding applications. Here, we report a systematic investigation of the size-dependent orthorhombic-to-tetragonal phase transition using a combined temperature-dependent optical, electrical transport and transmission electron microscopy study. Our studies of individual perovskite microplates with variable thicknesses demonstrate that the phase transition temperature decreases with reducing microplate thickness. The sudden decrease of mobility around phase transition temperature and the presence of hysteresis loops in the temperature-dependent mobility confirmmore » that the orthorhombic-to-tetragonal phase transition is a first-order phase transition. Lastly, our findings offer significant fundamental insight on the temperature-and size-dependent structural, optical and charge transport properties of perovskite materials, and can greatly impact future exploration of novel electronic and optoelectronic devices from these materials.« less

  14. A paper/polymer hybrid microfluidic microplate for rapid quantitative detection of multiple disease biomarkers

    PubMed Central

    Sanjay, Sharma T.; Dou, Maowei; Sun, Jianjun; Li, XiuJun

    2016-01-01

    Enzyme linked immunosorbent assay (ELISA) is one of the most widely used laboratory disease diagnosis methods. However, performing ELISA in low-resource settings is limited by long incubation time, large volumes of precious reagents, and well-equipped laboratories. Herein, we developed a simple, miniaturized paper/PMMA (poly(methyl methacrylate)) hybrid microfluidic microplate for low-cost, high throughput, and point-of-care (POC) infectious disease diagnosis. The novel use of porous paper in flow-through microwells facilitates rapid antibody/antigen immobilization and efficient washing, avoiding complicated surface modifications. The top reagent delivery channels can simply transfer reagents to multiple microwells thus avoiding repeated manual pipetting and costly robots. Results of colorimetric ELISA can be observed within an hour by the naked eye. Quantitative analysis was achieved by calculating the brightness of images scanned by an office scanner. Immunoglobulin G (IgG) and Hepatitis B surface Antigen (HBsAg) were quantitatively analyzed with good reliability in human serum samples. Without using any specialized equipment, the limits of detection of 1.6 ng/mL for IgG and 1.3 ng/mL for HBsAg were achieved, which were comparable to commercial ELISA kits using specialized equipment. We envisage that this simple POC hybrid microplate can have broad applications in various bioassays, especially in resource-limited settings. PMID:27456979

  15. Antimicrobial properties of cyclodextrin-antiseptics-complexes determined by microplate laser nephelometry and ATP bioluminescence assay.

    PubMed

    Finger, Susanne; Wiegand, Cornelia; Buschmann, Hans-Jürgen; Hipler, Uta-Christina

    2012-10-15

    Antimicrobial effects of substances can be determined with different methods that measure distinct parameters. Thus, a comparison of the results obtained can be difficult. In this study, two in vitro methods were employed to determine concentration and time dependent effects of cyclodextrin (CD)-complexes with the antiseptics chlorhexidine diacetate (CHX), iodine (IOD) and polihexanide (PHMB) on Candida albicans and Malassezia pachydermatis. Using both, microplate laser nephelometry and the ATP bioluminescence assay, it could be shown that CD-antiseptics-complexes tested exhibited significant antifungal effects with the exception of γ-CD-CHX in the case of C. albicans. Microplate laser nephelometry (MLN) is an optical method and enables a quantitative determination of particle concentrations in solution. By means of this method, microbial growth under influence of potential antimicrobial substances can be monitored over a prolonged time period. In addition, the antimicrobial activity was analyzed by measurement of the microbial adenosine triphosphate (ATP) content with a bioluminescent assay. The luminescent signal is directly proportional to the amount of ATP, and thus, a linear function of the number of living microbial cells present. Both methods were compared according to the half maximal inhibitory concentration (IC(50)) calculated and the statistical evaluation of Pearson's correlation coefficient (r). In summary, it could be demonstrated that both methods yield similar results although they differ in the parameter.

  16. Size-dependent phase transition in methylammonium lead iodide perovskite microplate crystals

    PubMed Central

    Li, Dehui; Wang, Gongming; Cheng, Hung-Chieh; Chen, Chih-Yen; Wu, Hao; Liu, Yuan; Huang, Yu; Duan, Xiangfeng

    2016-01-01

    Methylammonium lead iodide perovskite has attracted considerable recent interest for solution processable solar cells and other optoelectronic applications. The orthorhombic-to-tetragonal phase transition in perovskite can significantly alter its optical, electrical properties and impact the corresponding applications. Here, we report a systematic investigation of the size-dependent orthorhombic-to-tetragonal phase transition using a combined temperature-dependent optical, electrical transport and transmission electron microscopy study. Our studies of individual perovskite microplates with variable thicknesses demonstrate that the phase transition temperature decreases with reducing microplate thickness. The sudden decrease of mobility around phase transition temperature and the presence of hysteresis loops in the temperature-dependent mobility confirm that the orthorhombic-to-tetragonal phase transition is a first-order phase transition. Our findings offer significant fundamental insight on the temperature- and size-dependent structural, optical and charge transport properties of perovskite materials, and can greatly impact future exploration of novel electronic and optoelectronic devices from these materials. PMID:27098114

  17. A paper/polymer hybrid microfluidic microplate for rapid quantitative detection of multiple disease biomarkers

    NASA Astrophysics Data System (ADS)

    Sanjay, Sharma T.; Dou, Maowei; Sun, Jianjun; Li, Xiujun

    2016-07-01

    Enzyme linked immunosorbent assay (ELISA) is one of the most widely used laboratory disease diagnosis methods. However, performing ELISA in low-resource settings is limited by long incubation time, large volumes of precious reagents, and well-equipped laboratories. Herein, we developed a simple, miniaturized paper/PMMA (poly(methyl methacrylate)) hybrid microfluidic microplate for low-cost, high throughput, and point-of-care (POC) infectious disease diagnosis. The novel use of porous paper in flow-through microwells facilitates rapid antibody/antigen immobilization and efficient washing, avoiding complicated surface modifications. The top reagent delivery channels can simply transfer reagents to multiple microwells thus avoiding repeated manual pipetting and costly robots. Results of colorimetric ELISA can be observed within an hour by the naked eye. Quantitative analysis was achieved by calculating the brightness of images scanned by an office scanner. Immunoglobulin G (IgG) and Hepatitis B surface Antigen (HBsAg) were quantitatively analyzed with good reliability in human serum samples. Without using any specialized equipment, the limits of detection of 1.6 ng/mL for IgG and 1.3 ng/mL for HBsAg were achieved, which were comparable to commercial ELISA kits using specialized equipment. We envisage that this simple POC hybrid microplate can have broad applications in various bioassays, especially in resource-limited settings.

  18. Size-dependent phase transition in methylammonium lead iodide perovskite microplate crystals

    NASA Astrophysics Data System (ADS)

    Li, Dehui; Wang, Gongming; Cheng, Hung-Chieh; Chen, Chih-Yen; Wu, Hao; Liu, Yuan; Huang, Yu; Duan, Xiangfeng

    2016-04-01

    Methylammonium lead iodide perovskite has attracted considerable recent interest for solution processable solar cells and other optoelectronic applications. The orthorhombic-to-tetragonal phase transition in perovskite can significantly alter its optical, electrical properties and impact the corresponding applications. Here, we report a systematic investigation of the size-dependent orthorhombic-to-tetragonal phase transition using a combined temperature-dependent optical, electrical transport and transmission electron microscopy study. Our studies of individual perovskite microplates with variable thicknesses demonstrate that the phase transition temperature decreases with reducing microplate thickness. The sudden decrease of mobility around phase transition temperature and the presence of hysteresis loops in the temperature-dependent mobility confirm that the orthorhombic-to-tetragonal phase transition is a first-order phase transition. Our findings offer significant fundamental insight on the temperature- and size-dependent structural, optical and charge transport properties of perovskite materials, and can greatly impact future exploration of novel electronic and optoelectronic devices from these materials.

  19. Three Readers, Three Languages, Three Texts: The Strategic Reading of Multilingual and Multiliterate Readers

    ERIC Educational Resources Information Center

    Alsheikh, Negmeldin O.

    2011-01-01

    This case study investigates the metacognitive reading strategies of three advanced proficient trilingual readers whose native language is Hausa. The study examines the reading strategies employed by the three readers in English, French and Hausa. The aim of the study was to compare the reading strategy profiles of trilingual readers through…

  20. Reader-Centered Technical Writing

    NASA Astrophysics Data System (ADS)

    Narayanan, M.

    2012-12-01

    Technical writing is an essential part of professional communication and in recent years it has shifted from a genre-based approach. Formerly, technical writing primarily focused on generating templates of documents and sometimes it was creating or reproducing traditional forms with minor modifications and updates. Now, technical writing looks at the situations surrounding the need to write. This involves deep thinking about the goals and objectives of the project on hand. Furthermore, one observes that it is very important for any participatory process to have the full support of management. This support needs to be well understood and believed by employees. Professional writing may be very persuasive in some cases. When presented in the appropriate context, technical writing can persuade a company to improve work conditions ensuring employee safety and timely production. However, one must recognize that lot of professional writing still continues to make use of reports and instruction manuals. Normally, technical and professional writing addresses four aspects. Objective: The need for generating a given professionally written technical document and the goals the document is expected to achieve and accomplish. Clientele: The clientele who will utilize the technical document. This may include the people in the organization. This may also include "unintended readers." Customers: The population that may be affected by the content of the technical document generated. This includes the stakeholders who will be influenced. Environment: The background in which the document is created. Also, the nature of the situation that warranted the generation of the document. Swiss Psychologist Jean Piaget's view of Learning focuses on three aspects. The author likes to extend Jean Piaget's ideas to students, who are asked to prepare and submit Reader-Centered Technical Writing reports and exercises. Assimilation: Writers may benefit specifically, by assimilating a new object into

  1. Notebook Connections: Strategies for the Reader's Notebook

    ERIC Educational Resources Information Center

    Buckner, Aimee

    2009-01-01

    In "Notebook Know-How", Aimee Buckner demonstrated the power of notebooks to spark and capture students' ideas in the writing workshop. In "Notebook Connections", she turns her focus to the reading workshop, showing how to transform those "couch-potato" readers into deep thinkers. Buckner's fourth-grade students use reader's notebooks as a place…

  2. Designing Online Courses for Screen Reader Users

    ERIC Educational Resources Information Center

    Kearns, Lorna R.; Frey, Barbara A.; McMorland, Gabriel

    2013-01-01

    A review of multiple online courses at one institution was conducted by a skilled screen reader user for the purpose of assessing the extent to which the courses were navigable and understandable to online students using assistive technologies. This paper identifies features of online courses that may present problems for screen reader users and…

  3. Hemispheric Specialization in Normal and Disabled Readers.

    ERIC Educational Resources Information Center

    Newell, David; Rugel, Robert P.

    1981-01-01

    Degree of hemispheric specialization in 32 normal and 32 disabled readers (9 to 12 years old) was evaluated using a linguistic task and a musical task. While normals demonstrated hemispheric specialization on these tasks in the expected directions, disabled readers showed a right hemisphere deficit. (Author)

  4. Planning Behaviour in Good and Poor Readers

    ERIC Educational Resources Information Center

    Mahapatra, Shamita

    2016-01-01

    A group of 50 good readers and a group of 50 poor readers of Grade 5 matched for age and intelligence and selected on the basis of their proficiency in reading comprehension were tested for their competence in word reading and the process of planning at three different levels, namely, perceptual, memory and conceptual in order to study the…

  5. Fiction Reading Strategies of College Readers

    ERIC Educational Resources Information Center

    Poole, Alex

    2013-01-01

    Research has shown that skilled college readers use a variety of strategies in flexible ways in order to comprehend academic texts. However, little is known about how skilled college readers use strategies when reading fiction, in spite of the fact that literature courses are required at many universities and thousands of students regularly major…

  6. THE LAY READER PROGRAM IN ACTION.

    ERIC Educational Resources Information Center

    BURKE, VIRGINIA M.

    IN 1960, THE WISCONSIN COUNCIL OF TEACHERS OF ENGLISH CONDUCTED AN INTERVIEW AND QUESTIONNAIRE SURVEY OF PARTICIPANTS IN TWO LAY-READER PROGRAMS AT THE SECONDARY LEVEL--(1) A PROGRAM AT RACINE IN ITS FIRST FULL SEMESTER IN WHICH LAY READERS CORRECTED AND EVALUATED, BUT DID NOT GRADE, APPROXIMATELY HALF OF THE THEMES FROM SELECTED CLASSES, AND (2)…

  7. Attracting New Readers with Hip Hop Lit

    ERIC Educational Resources Information Center

    Meloni, Christine

    2007-01-01

    With the explosion of technology, librarians are not the only ones who sometimes feel that reading has taken a backseat as a recreational activity. Readers are not reading like they used to. However, while overall readership is down in almost every demographic group, African-Americans, a faction traditionally considered reluctant readers, are…

  8. Helping Your Child Become a Reader

    ERIC Educational Resources Information Center

    US Department of Education, 2008

    2008-01-01

    "Helping Your Child Become a Reader" includes information about why and how to use language skills (talking and listening, reading, and writing) to help young children grow into readers. Everyday activities and a list of resources for parents to encourage children's love of reading and strengthen language skills are also provided. This brochure…

  9. High Ability Readers and the Achievement Gap

    ERIC Educational Resources Information Center

    Hunsaker, Scott L.; Parke, Cynthia J.; Bramble, Joan G.

    2004-01-01

    To close the achievement gap, the "No Child Left Behind" law calls for all students to make appropriate yearly progress. This presumably means that progress is being made by capable readers at the same time progress is being made by struggling readers. However, there appear to be unintended effects of "No Child Left Behind"…

  10. Intermediate Amharic Cultural Reader. Final Report.

    ERIC Educational Resources Information Center

    Leslau, Wolf

    This reader is intended to provide material for the intermediate-level student of Amharic, as well as to introduce the student to the cultural and social life of Ethiopia. The 39 texts were each prepared by a different student at Haile Selassie I University, thus providing the reader with a variety of language styles. The Amharic texts are…

  11. Reader Response in the Teaching of Composition.

    ERIC Educational Resources Information Center

    Price, Marian

    As an interesting development in recent literary criticism, reader response can enhance a composition class in many ways. Reader response, by incorporating both intellect and feeling into an aesthetic reaction to literature, restores the subjective aspect that some forms of criticism deny. Three main elements compose the repeated cycle of a…

  12. Developing Readers: Lessons from Agatha Christie.

    ERIC Educational Resources Information Center

    Mackey, Margaret

    1991-01-01

    Reminds readers that looking at individual authors rather than dealing in generalizations is a very meaningful approach to increasing an understanding of how reading works by making sense of the text. Discusses how Agatha Christie's writing follows very predictable patterns and may provide useful experiences for novice readers. (MG)

  13. Readers Theatre News, 1974 Bibliography Issue.

    ERIC Educational Resources Information Center

    Barnhart, Alice, Ed.

    The contents of this issue consist of two articles, Readers Theatre as a Humanizing Process by Wallace Bacon and The Secret of Individual Progress in Art by Paul Gregory, and the 1974 readers theatre bibliography. Items are listed in the bibliography in four categories: plays, poetry, programs, and prose. In addition, the dates and places of…

  14. Enhancing Memory Access for Less Skilled Readers

    ERIC Educational Resources Information Center

    Smith, Emily R.; O'Brien, Edward J.

    2016-01-01

    Less skilled readers' comprehension often suffers because they have an impoverished representation of text in long-term memory; this, in turn, increases the difficulty of gaining access to backgrounded information necessary for maintaining coherence. The results of four experiments demonstrated that providing less skilled readers with additional…

  15. Readers Theatre: An Introduction to Classroom Performance.

    ERIC Educational Resources Information Center

    Ratliff, Gerald Lee

    One of the primary principles of Readers Theater is to "dramatize" literature in classroom performance and to provide a visual and oral stimulus to students who are unaccustomed to using imagination to appreciate literary texts. Readers Theater may be used to enhance the critical study of language; to explore author meaning or point of view; to…

  16. Professional Roles: Inventing Writers for Imagined Readers.

    ERIC Educational Resources Information Center

    Meese, George P. E.

    Careful audience analysis should be a major focus of the rhetorical side of technical writing courses for university undergraduates. Student writers need to be taught to appreciate their readers' problems, interests, and motives. Most often, audience analysis is accomplished imaginatively--authors create mental images of their readers. Students…

  17. KURDISH READERS. PART II, KURDISH ESSAYS.

    ERIC Educational Resources Information Center

    ABDULLA, JAMAL JALAL; MCCARUS, ERNEST N.

    THIS READER, TOGETHER WITH THE "NEWSPAPER KURDISH" AND "SHORT STORIES" READERS, FOLLOWS THE "BASIC COURSE IN KURDISH" (BY THE SAME AUTHORS) AND ASSUMES A MASTERY OF THE BASIC ELEMENTS OF THE STRUCTURE AND WRITING SYSTEM AS PRESENTED IN THE BEGINNING COURSE. WRITTEN IN THE SULAIMANIAN DIALECT, THE OFFICIAL DIALECT OF…

  18. Using Readers' Theater with Multicultural Literature

    ERIC Educational Resources Information Center

    Weisenburger, Stephanie

    2009-01-01

    The author needed a way to engage her students in the reading process and found one extremely successful strategy: using Readers' Theater. Readers' Theater "dramatizes" literature through a classroom performance and provides visual and oral stimulus to students who are not used to using imagination to appreciate literary texts. It involves a…

  19. Reader Imposed Structure and Prose Retention.

    ERIC Educational Resources Information Center

    Grabe, Mark D.

    1979-01-01

    Two experiments investigated the impact of a reader's perspective on prose learning: (1) subjects read stories from one of two directed perspectives or with no directed perspective; or (2) readers organized and familiarized themselves with a perspective before the perspective was applied to a story. Perspective influenced recall and organization.…

  20. Developing Preservice Teachers' Perspectives on Reader Response.

    ERIC Educational Resources Information Center

    Grisham, Dana L.

    2001-01-01

    Examines preservice teachers' developing conceptions of reader response theory, specifically focusing on the importance of aesthetic response to students' engagement with and motivation for reading. Finds that the aesthetic reader stance predominated in students' written responses and discussions; and that written response did not influence the…

  1. Using Nooks to Hook Reluctant Readers

    ERIC Educational Resources Information Center

    Dierking, Rebecca

    2015-01-01

    This article presents the findings of a two-year qualitative study of electronic reading device use with high school sophomores, most of whom self-identified as reluctant or struggling readers. Electronic readers were used primarily in one weekly fifty-minute class period, during silent sustained reading, wherein students chose freely their texts.…

  2. From a collage of microplates to stable continental crust - an example from Precambrian Europe

    NASA Astrophysics Data System (ADS)

    Korja, Annakaisa

    2013-04-01

    Svecofennian orogen (2.0-1.7 Ga) comprises the oldest undispersed orogenic belt on Baltica and Eurasian plate. Svecofennian orogenic belt evolved from a series of short-lived terrane accretions around Baltica's Archean nucleus during the formation of the Precambrian Nuna supercontinent. Geological and geophysical datasets indicate W-SW growth of Baltica with NE-ward dipping subduction zones. The data suggest a long-lived retreating subduction system in the southwestern parts whereas in the northern and central parts the northeasterly transport of continental fragments or microplates towards the continental nucleus is also documented. The geotectonic environment resembles that of the early stages of the Alpine-Himalayan or Indonesian orogenic system, in which dispersed continental fragments, arcs and microplates have been attached to the Eurasian plate margin. Thus the Svecofennian orogeny can be viewed as proxy for the initial stages of an internal orogenic system. Svecofennian orogeny is a Paleoproterozoic analogue of an evolved orogenic system where terrane accretion is followed by lateral spreading or collapse induced by change in the plate architecture. The exposed parts are composed of granitoid intrusions as well as highly deformed supracrustal units. Supracrustal rocks have been metamorphosed in LP-HT conditions in either paleo-lower-upper crust or paleo-upper-middle crust. Large scale seismic reflection profiles (BABEL and FIRE) across Baltica image the crust as a collage of terranes suggesting that the bedrock has been formed and thickened in sequential accretions. The profiles also image three fold layering of the thickened crust (>55 km) to transect old terrane boundaries, suggesting that the over-thickened bedrock structures have been rearranged in post-collisional spreading and/or collapse processes. The middle crust displays typical large scale flow structures: herringbone and anticlinal ramps, rooted onto large scale listric surfaces also suggestive

  3. Localized deformation zones in the offshore leading edge of the Yakutat microplate, Gulf of Alaska

    NASA Astrophysics Data System (ADS)

    Lowe, L. A.; Gulick, S. P.; Pavlis, T.; Bruhn, R. L.; Mann, P.

    2006-12-01

    The Gulf of Alaska margin is dominated by the collision and subduction of the Yakutat microplate as it travels northwest with respect to North America at near Pacific Plate velocities (\\~45 mm/yr). The oblique Yakutat block collision with North America is in transition between convergence to the west and translation along the Queen Charlotte-Fairweather-Denali Fault system to the east and north. Industry seismic reflection and high- resolution seismic reflection data collected by the R/V Maurice Ewing (2004) provides insight into how the Yakutat-North America collision is accommodated by active offshore structures near the leading edge of the Yakutat microplate. A \\~200 km wide area bounded by the Ten Fathom Fault, the offshore N. America-Yakutat contact, to the west and the eastern edge of the Pamplona Zone (PZ) to the east has previously been mapped as a continuous deformation zone consisting of NE-SW trending imbricate thrusts and folds. Though this mapping corroborates onshore measurements of active deformation west of the Bering Glacier in the Yakutat block, the relationship between current onshore deformation and the observed offshore structures remains unclear. Our observations indicate that neotectonic deformation is accommodated offshore by highly localized, asynchronous thrusts that, when analyzed in an accretionary context, may be connected by a sub-horizontal decollement. Data from the eastern edge of the PZ, the proposed deformation front, shows surface deformation caused by east-verging thrust faults. Seismic reflection profiles in the western PZ and the Bering Trough show no evidence of active tectonic deformation and up to \\~200 m of undisturbed sediments indicating that faulting in this part of the Yakutat block has been inactive since the Last Glacial Maximum or earlier. Farther west, above the Kayak Island fault zone, directly east of the Ten Fathom Fault, the presence of up to \\~50 m of undeformed sediments suggests a recent (ca. 14 ka

  4. Microplate-based active/inactive 1 screen for biomass degrading enzyme library purification and gene discovery

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We present here a whole-cell and permeabilized E. coli cell 1' active/inactive microplate screen for ß-D-xylosidase, xylanase, endocellulase, and ferulic acid esterase enzyme activities which are critical for the enzymatic deconstruction of biomass for fuels and chemicals. Transformants from genomic...

  5. An acquired distaste: Sugar discrimination by the larval parasitoid Microplitis croceipes (Hymenoptera: Braconidae) is affected by prior sugar exposure

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As sugar quality feeding is very important in the lives of adult parasitoids, we examined several feeding responses of Microplitis croceipes to sugars commonly found in nectar. We first examined the relationship between feeding time and consumption of sucrose, glucose, fructose and maltose by Microp...

  6. The role of photoperiod and temperature in diapause induction of the endoparasitoid wasp, Microplitis mediator (Haliday) (Hymenoptera: Braconidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Diapause in Microplitis mediator is manifested during the pupal stage and normally occurs during the winter season because of a photoperiodic response which is highly dependant on temperature. In the reported study, diapause was determine by photoperiod and mediated by temperature, which supports th...

  7. Microplate assay analysis of the distribution of organophosphate and carbamate resistance in Guatemalan Anopheles albimanus

    PubMed Central

    Brogdon, W. G.; Beach, R. F.; Stewart, J. M.; Castanaza, L.

    1988-01-01

    Simple microplate assay methods for determining the frequency of insecticide resistance in single mosquitos were used to study the distribution and localization of organophosphate and carbamate resistance in field populations of Anopheles albimanus Weidemann in Guatemala, where such resistance, caused by heavy use of agricultural pesticides, has long been assumed to be widespread. Areas of complete susceptibility to organophosphates and carbamates were observed, as well as areas where the resistant phenotypes represented up to 98% of the population. Overall, the resistance levels were lower and more localized than expected. Two mechanisms of resistance were identified by the microassay methods. These were the elevated esterase (nonspecific esterase) and insensitive acetylcholinesterase mechanisms which were selected independently, the former (documented for the first time in Central American anophelines) being predominant. These methods represent a promising new technology for the detection and assessment of resistance and will facilitate improved control strategy decisions. PMID:3262440

  8. Rapid microplate, green method for high-throughput evaluation of vinegar acidity using thermal infrared enthalpimetry.

    PubMed

    Tischer, Bruna; Oliveira, Alessandra Stangherlin; Ferreira, Daniele de Freitas; Menezes, Cristiano Ragagnin; Duarte, Fábio Andrei; Wagner, Roger; Barin, Juliano Smanioto

    2017-01-15

    Infrared thermal imaging was combined with disposable microplates to perform enthalpimetric analysis using an infrared camera to monitor temperature without contact. The proposed thermal infrared enthalpimetry (TIE) method was used to determine the total, fixed and volatile acidities of vinegars. Sample preparation and analysis were performed in the same vessel, avoiding excessive sample handling and reducing energy expenditure by more than ten times. The results agreed with those of the conventional method for different kinds of vinegars, with values of 1.7%, and 2.3% for repeatability and intermediate precision, respectively. A linear calibration curve was obtained from 0.040 to 1.30molL(-1). The proposed method provided rapid results (within 10s) for four samples simultaneously, a sample throughput of up to 480 samples per hour. In addition, the method complies with at least eight of twelve recommendations for green analytical chemistry, making TIE a promising tool for routine vinegar analysis.

  9. Modal interactions in primary and subharmonic resonant dynamics of imperfect microplates with geometric nonlinearities

    NASA Astrophysics Data System (ADS)

    Farokhi, Hamed; Ghayesh, Mergen H.

    2016-06-01

    This paper analyses the modal interactions in the nonlinear, size-dependent dynamics of geometrically imperfect microplates. Based on the modified couple stress theory, the equations of motion for the in-plane and out-of-plane motions are obtained employing the von Kármán plate theory as well as Kirchhoff's hypotheses by means of the Lagrange equations. The equations of motions are solved using the pseudo-arclength continuation technique and direct time-integration method. The system parameters are tuned to the values associated with modal interactions, and then nonlinear resonant responses and energy transfer are analysed. Nonlinear motion characteristics are shown in the form of frequency-response and force-response curves, time histories, phase-plane portraits, and fast Fourier transforms.

  10. Perceiving the epigenetic landscape through histone readers

    PubMed Central

    Musselman, Catherine A.; Lalonde, Marie-Eve; Côté, Jacques; Kutateladze, Tatiana G.

    2013-01-01

    Post-translational modifications (PTMs) of histones provide a fine-tuned mechanism for regulating chromatin structure and dynamics. PTMs can alter direct interactions between histones and DNA and serve as docking sites for protein effectors, or readers, of these PTMs. Binding of the readers recruits or stabilizes various components of the nuclear signaling machinery at specific genomic sites, mediating fundamental DNA-templated processes, including gene transcription and DNA recombination, replication and repair. In this review, we highlight the latest advances in characterizing histone-binding mechanisms and identifying new epigenetic readers and summarize the functional significance of PTM recognition. PMID:23211769

  11. Deep sequencing identifies viral and wasp genes with potential roles in replication of Microplitis demolitor Bracovirus.

    PubMed

    Burke, Gaelen R; Strand, Michael R

    2012-03-01

    Viruses in the genus Bracovirus (BV) (Polydnaviridae) are symbionts of parasitoid wasps that specifically replicate in the ovaries of females. Recent analysis of expressed sequence tags from two wasp species, Cotesia congregata and Chelonus inanitus, identified transcripts related to 24 different nudivirus genes. These results together with other data strongly indicate that BVs evolved from a nudivirus ancestor. However, it remains unclear whether BV-carrying wasps contain other nudivirus-like genes and what types of wasp genes may also be required for BV replication. Microplitis demolitor carries Microplitis demolitor bracovirus (MdBV). Here we characterized MdBV replication and performed massively parallel sequencing of M. demolitor ovary transcripts. Our results indicated that MdBV replication begins in stage 2 pupae and continues in adults. Analysis of prereplication- and active-replication-stage ovary RNAs yielded 22 Gb of sequence that assembled into 66,425 transcripts. This breadth of sampling indicated that a large percentage of genes in the M. demolitor genome were sequenced. A total of 41 nudivirus-like transcripts were identified, of which a majority were highly expressed during MdBV replication. Our results also identified a suite of wasp genes that were highly expressed during MdBV replication. Among these products were several transcripts with conserved roles in regulating locus-specific DNA amplification by eukaryotes. Overall, our data set together with prior results likely identify the majority of nudivirus-related genes that are transcriptionally functional during BV replication. Our results also suggest that amplification of proviral DNAs for packaging into BV virions may depend upon the replication machinery of wasps.

  12. Rigidity of Major Plates and Microplates Estimated From GPS Solution GPS2006.0

    NASA Astrophysics Data System (ADS)

    Kogan, M. G.; Steblov, G. M.

    2006-05-01

    Here we analyze the rigidity of eight major lithospheric plates using our global GPS solution GPS2006.0. We included all daily observations in interval 1995.0 to 2006.0 collected at IGS stations, as well as observations at many important stations not included in IGS. Loose multiyear solution GPS2006.0 is based on daily solutions by GAMIT software, performed at SOPAC and at Columbia University; those daily solutions were combined by Kalman filter (GLOBK software) into a loose multiyear solution. The constrained solution for station positions and velocities was obtained without a conventional reference frame; instead, we applied translation and rotation in order to best fit the zero velocities of 76 stations in stable plate cores excluding the regions of postglacial rebound. Simultaneously, we estimated relative plate rotation vectors (RV) and the origin translation rate (OTR), and then corrected station velocities for it. Therefore, the velocities in GPS2006.0 are unaffected by the OTR error of ITRF2000 conventionally used to constrain a loose solution. The 1-sigma plate-residual velocity in a stable plate core is less than 1 mm/yr for the plates: Eurasia, Pacific, North and South Americas, Nubia, Australia, and Antarctica; it is 1.4 mm/yr for the Indian plate, most probably because of poorer data quality. Plate-residuals at other established plates (Arabia, Nazca, Caribbean, Philippine) were not assessed for lack of observations. From our analysis, an upper bound for the mobility of the plate inner area is 1 mm/yr. Plate- residual GPS velocities for several hypothesized microplates in east Asia, such as Okhotsk, Amuria, South China, are 3-4 times higher; corresponding strain rates for these microplates are an order of magnitude higher than for Eurasia, North America, and other large plates.

  13. Deformation driven by subduction and microplate collision: Geodynamics of Cook Inlet basin, Alaska

    USGS Publications Warehouse

    Bruhn, R.L.; Haeussler, P.J.

    2006-01-01

    Late Neogene and younger deformation in Cook Inlet basin is caused by dextral transpression in the plate margin of south-central Alaska. Collision and subduction of the Yakutat microplate at the northeastern end of the Aleutian subduction zone is driving the accretionary complex of the Chugach and Kenai Mountains toward the Alaska Range on the opposite side of the basin. This deformation creates belts of fault-cored anticlines that are prolific traps of hydrocarbons and are also potential sources for damaging earthquakes. The faults dip steeply, extend into the Mesozoic basement beneath the Tertiary basin fill, and form conjugate flower structures at some localities. Comparing the geometry of the natural faults and folds with analog models created in a sandbox deformation apparatus suggests that some of the faults accommodate significant dextral as well as reverse-slip motion. We develop a tectonic model in which dextral shearing and horizontal shortening of the basin is driven by microplate collision with an additional component of thrust-type strain caused by plate subduction. This model predicts temporally fluctuating stress fields that are coupled to the recurrence intervals of large-magnitude subduction zone earthquakes. The maximum principal compressive stress is oriented east-southeast to east-northeast with nearly vertical least compressive stress when the basin's lithosphere is mostly decoupled from the underlying subduction megathrust. This stress tensor is compatible with principal stresses inferred from focal mechanisms of earthquakes that occur within the crust beneath Cook Inlet basin. Locking of the megathrust between great magnitude earthquakes may cause the maximum principal compressive stress to rotate toward the northwest. Moderate dipping faults that strike north to northeast may be optimally oriented for rupture in the ambient stress field, but steeply dipping faults within the cores of some anticlines are unfavorably oriented with respect to

  14. Crustal Structure and Deformation of the Yakutat Microplate: New Insights From STEEP Marine Seismic Reflection Data

    NASA Astrophysics Data System (ADS)

    Lowe, L. A.; Gulick, S. P.; Christeson, G.; van Avendonk, H.; Reece, R.; Elmore, R.; Pavlis, T.

    2008-12-01

    In fall 2008, we will conduct an active source marine seismic experiment of the offshore Yakutat microplate in the northern Gulf of Alaska. The survey will be conducted aboard the academic research vessel, R/V Marcus Langseth, collecting deep-penetrating multi-channel seismic reflection survey using an 8-km, 640 channel hydrophone streamer and a 6600 cu. in., 36 airgun array. The survey is the concluding data acquisition phase for the ST. Elias Erosion and tectonics Project (STEEP), a multi-institution NSF-Continental Dynamics project investigating the interplay of climate and tectonics in the Chugach-St. Elias Mountains in southern Alaska. The experiment will also provide important site survey information for possible future Integrated Ocean Drilling Program investigations. Two profiles coincident with wide-angle refraction data (see Christeson, et al., this session) will image structural changes across the Dangerous River Zone from east to west and the Transition Fault from south to north. We will also image the western portion of the Transition Fault to determine the nature of faulting along this boundary including whether or not the Pacific Plate is underthrusting beneath the Yakutat microplate as part of this collision. Our westernmost profile will image the Kayak Island Zone, typically described as the northern extension of the Aleutian megathrust but which may be a forming suture acting as a deformation backstop for the converging Yakutat and North American plates. Profiles across the Pamplona Zone, the current Yakutat-North America deformation front, will further constrain relative timing of structural development and the depth of deformation on the broad folds and thrust faults that comprise the area. This new dataset will allow further insight into regional tectonics of the St. Elias region as well as provide more detail regarding the development of the south Alaskan margin during major Plio-Pleistocene glacial- interglacial periods.

  15. Reader Profiles for Adults with Low Literacy Skills: A Quest to Find Resilient Readers

    PubMed Central

    Binder, Katherine S.; Lee, Cheryl; College, Mount Holyoke

    2014-01-01

    Resilient readers are those who, despite their poor phonological decoding skills, have good comprehension abilities (Jackson & Doellinger, 2002). Thus far, these readers have been identified in college settings. The purpose of this study was to a) determine if this reader profile was present in a sample taken from an Adult Basic Education (ABE) population, and b) identify compensatory mechanisms these readers might use to better their reading comprehension. We administered a battery of tasks consisting of non-word reading, comprehension, fluency, and orthographic processing to a diverse sample of adults in ABE classes. Not only did we identify a group of resilient readers in this sample, we identified three other sub-groups: unskilled readers who had poor decoding and comprehension abilities, skilled readers who possessed good decoding and comprehension abilities, and a group of individuals who had good decoding skills but poor comprehension abilities. We found that the resilient readers and good decoders/poor comprehenders had better orthographic and fluency skills compared to the unskilled readers. However, these last two groups produced different error patterns on the orthographic and fluency tasks. We discuss the implications that these very different reader profiles have for ABE programs. PMID:25431747

  16. Turning Polite Guests into Executive Readers.

    ERIC Educational Resources Information Center

    Gates, Dale D.

    1983-01-01

    Expands the usual notion of critical reading from just the judgments readers make about the text to include judgments they make about their "reading" of the text. Describes activities to develop "executive" metacomprehension skills. (HTH)

  17. "Guy Reader" Is Not an Oxymoron

    ERIC Educational Resources Information Center

    Korman, Gordon

    2013-01-01

    The author of 80 books, most recently "HYPNOTIZE ME," discusses the unique challenges of engaging reluctant and sometimes openly hostile boys by creating the kind of stories that will hook them in and turn them into lifelong readers.

  18. 38 CFR 21.150 - Reader service.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... which the veteran has been furnished to enable him or her to read printed material unassisted. (e) Recording. VA will not normally pay for recording textbooks or other materials as a part of reader...

  19. 38 CFR 21.150 - Reader service.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... which the veteran has been furnished to enable him or her to read printed material unassisted. (e) Recording. VA will not normally pay for recording textbooks or other materials as a part of reader...

  20. Finding Hooks to Catch Reluctant Readers.

    ERIC Educational Resources Information Center

    Grimes, Marijo

    1991-01-01

    Discusses a list of books of varying lengths and difficulty that the reading teacher can use to help reluctant readers find that all-important breakthrough book to discover what reading can do for them. (KEH)

  1. Surface oxide effect on optical sensing and photoelectric conversion of α-In2Se3 hexagonal microplates.

    PubMed

    Ho, Ching-Hwa; Lin, Chien-Hao; Wang, Yi-Ping; Chen, Ying-Cen; Chen, Shin-Hong; Huang, Ying-Sheng

    2013-03-01

    The surface formation oxide assists of visible to ultraviolet photoelectric conversion in α-In2Se3 hexagonal microplates has been explored. Hexagonal α-In2Se3 microplates with the sizes of 10s to 100s of micrometers were synthesized and prepared by the chemical vapor transport method using ICl3 as a transport agent. Many vacancies and surface imperfection states have been found in the bulk and on the surface of the microplate because of the intrinsic defect nature of α-In2Se3. To discover physical and chemical properties and finding technological uses of α-In2Se3, several experiments including transmission electron miscopy (TEM), X-ray photoelectron spectroscopy (XPS), surface photovoltage (SPV), photoluminescence (PL), surface photoresponse (SPR), photoconductivity (PC), and thermoreflectance (TR) measurements have been carried out. Experimental results of TEM, XPS, SPV, PL, and SPR measurements show that a surface oxidation layer α-In2Se3-3xO3x (0 ≤ x ≤ 1) has formed on the crystal face of α-In2Se3 in environmental air with the inner layer content close to In2Se3 but the outermost layer content approaching In2O3. The near band edge transitions of α-In2Se3 microplates have been probed experimentally by TR and PC measurements. The direct band gap of α-In2Se3 has been determined to be 1.453 eV. The SPV result shows a maximum quantum efficiency of the surface oxide α-In2Se3-3xO3x (0 ≤ x ≤ 1) that presents a peak photoresponse near 2.18 eV. The analyses of SPV, SPR, PL, TR, and PC measurements revealed that the surface oxide layer facilitates the conversion of the ultraviolet to the visible range while the native defects (Se and In vacancies) sustain photoconductivity in the near-infrared region. On the basis of the experimental results a wide-energy-range photodetector that combines PC- and SPR-mode operations for α-In2Se3 microplate has been made. The testing results show a well-behaved function of photoelectric conversion in the near-infrared to

  2. Fluorescence lifetime plate reader: resolution and precision meet high-throughput.

    PubMed

    Petersen, Karl J; Peterson, Kurt C; Muretta, Joseph M; Higgins, Sutton E; Gillispie, Gregory D; Thomas, David D

    2014-11-01

    We describe a nanosecond time-resolved fluorescence spectrometer that acquires fluorescence decay waveforms from each well of a 384-well microplate in 3 min with signal-to-noise exceeding 400 using direct waveform recording. The instrument combines high-energy pulsed laser sources (5-10 kHz repetition rate) with a photomultiplier and high-speed digitizer (1 GHz) to record a fluorescence decay waveform after each pulse. Waveforms acquired from rhodamine or 5-((2-aminoethyl)amino) naphthalene-1-sulfonic acid dyes in a 384-well plate gave lifetime measurements 5- to 25-fold more precise than the simultaneous intensity measurements. Lifetimes as short as 0.04 ns were acquired by interleaving with an effective sample rate of 5 GHz. Lifetime measurements resolved mixtures of single-exponential dyes with better than 1% accuracy. The fluorescence lifetime plate reader enables multiple-well fluorescence lifetime measurements with an acquisition time of 0.5 s per well, suitable for high-throughput fluorescence lifetime screening applications.

  3. Damping constant estimation in magnetoresistive readers

    SciTech Connect

    Stankiewicz, Andrzej Hernandez, Stephanie

    2015-05-07

    The damping constant is a key design parameter in magnetic reader design. Its value can be derived from bulk or sheet film ferromagnetic resonance (FMR) line width. However, dynamics of nanodevices is usually defined by presence of non-uniform modes. It triggers new damping mechanisms and produces stronger damping than expected from traditional FMR. This work proposes a device-level technique for damping evaluation, based on time-domain analysis of thermally excited stochastic oscillations. The signal is collected using a high bandwidth oscilloscope, by direct probing of a biased reader. Recorded waveforms may contain different noise signals, but free layer FMR is usually a dominating one. The autocorrelation function is a reflection of the damped oscillation curve, averaging out stochastic contributions. The damped oscillator formula is fitted to autocorrelation data, producing resonance frequency and damping constant values. Restricting lag range allows for mitigation of the impact of other phenomena (e.g., reader instability) on the damping constant. For a micromagnetically modeled reader, the technique proves to be much more accurate than the stochastic FMR line width approach. Application to actual reader waveforms yields a damping constant of ∼0.03.

  4. Examining Readers' Evaluations of Objectivity and Bias in News Discourse

    ERIC Educational Resources Information Center

    Cramer, Peter; Eisenhart, Christopher

    2014-01-01

    Readers' objectivity and bias evaluations of news texts were investigated in order to better understand the process by which readers make these kinds of judgments and the evidence on which they base them. Readers were primed to evaluate news texts for objectivity and bias, and their selections and metacommentary were analyzed. Readers detected…

  5. Reader Response Theory in the High School English Classroom.

    ERIC Educational Resources Information Center

    Shelton, Karen Yvonne

    A study examined the theory concerning reader response and the rationale and practice of reader response in the high school English curriculum. Formal experimental studies existed that explored reader response practices in the high school setting, but no formal studies existed on the questioning practices of potential reader response teachers. A…

  6. The Effect of Thermal Annealing on Charge Transport in Organolead Halide Perovskite Microplate Field-Effect Transistors.

    PubMed

    Li, Dehui; Cheng, Hung-Chieh; Wang, Yiliu; Zhao, Zipeng; Wang, Gongming; Wu, Hao; He, Qiyuan; Huang, Yu; Duan, Xiangfeng

    2017-01-01

    Transformation of unipolar n-type semiconductor behavior to ambipolar and finally to unipolar p-type behavior in CH3 NH3 PbI3 microplate field-effect transistors by thermal annealing is reported. The photoluminescence spectra essentially maintain the same features before and after the thermal annealing process, demonstrating that the charge transport measurement provides a sensitive way to probe low-concentration defects in perovskite materials.

  7. "It's All Coming Together": An Encounter between Implied Reader and Actual Reader in the Australian Rainforest

    ERIC Educational Resources Information Center

    Williams, Sandra J.

    2008-01-01

    In this paper I discuss how taking a particular literary theory--the implied reader--serves to offer a focus for the teacher's initial reading of a text and provides a formative assessment tool. Iser's Implied Reader theory is discussed, after which a picture book, "Where the Forest Meets the Sea" by Jeannie Baker, is analysed from this…

  8. A SURVEY OF MICROFICHE READERS AND READER-PRINTERS CURRENTLY MANUFACTURED IN THE UNITED STATES.

    ERIC Educational Resources Information Center

    TATE, VERNON D.; WOLF, DAVID R.

    A DISCUSSION OF MICROFICHE, INCLUDING DEFINITION OF THE TERM, CONSIDERATION OF ITS CHARACTERISTICS AND DIFFERENCES BETWEEN IT AND OTHER MICROFORMS, PUBLICATION IN THE FORM, AND STANDARDIZATION OF ITS SIZE AND FORMAT IS FOLLOWED BY AN EXPLANATION OF THE SURVEY OF MICROFICHE READERS AND READER-PRINTERS. THE SURVEY ITSELF GIVES THE FOLLOWING…

  9. Breaking off Engagement: Readers' Disengagement as a Function of Reader and Text Characteristics

    ERIC Educational Resources Information Center

    Goedecke, Patricia J.; Dong, Daqi; Shi, Genghu; Feng, Shi; Risko, Evan; Olney, Andrew M.; D'Mello, Sidney K.; Graesser, Arthur C.

    2015-01-01

    Engagement during reading can be measured by the amount of time readers invest in the reading process. It is hypothesized that disengagement is marked by a decrease in time investment as compared with the demands made on the reader by the text. In this study, self-paced reading times for screens of text were predicted by a text complexity score…

  10. Peptide code-on-a-microplate for protease activity analysis via MALDI-TOF mass spectrometric quantitation.

    PubMed

    Hu, Junjie; Liu, Fei; Ju, Huangxian

    2015-04-21

    A peptide-encoded microplate was proposed for MALDI-TOF mass spectrometric (MS) analysis of protease activity. The peptide codes were designed to contain a coding region and the substrate of protease for enzymatic cleavage, respectively, and an internal standard method was proposed for the MS quantitation of the cleavage products of these peptide codes. Upon the cleavage reaction in the presence of target proteases, the coding regions were released from the microplate, which were directly quantitated by using corresponding peptides with one-amino acid difference as the internal standards. The coding region could be used as the unique "Protease ID" for the identification of corresponding protease, and the amount of the cleavage product was used for protease activity analysis. Using trypsin and chymotrypsin as the model proteases to verify the multiplex protease assay, the designed "Trypsin ID" and "Chymotrypsin ID" occurred at m/z 761.6 and 711.6. The logarithm value of the intensity ratio of "Protease ID" to internal standard was proportional to trypsin and chymotrypsin concentration in a range from 5.0 to 500 and 10 to 500 nM, respectively. The detection limits for trypsin and chymotrypsin were 2.3 and 5.2 nM, respectively. The peptide-encoded microplate showed good selectivity. This proposed method provided a powerful tool for convenient identification and activity analysis of multiplex proteases.

  11. Poor Readers of Chinese Respond Slower than Good Readers in Phonological, Rapid Naming, and Interval Timing Tasks

    ERIC Educational Resources Information Center

    Penney, Trevor B.; Leung, Kar Man; Chan, Po Chi; Meng, Xiangzhi; McBride-Chang, Catherine A.

    2005-01-01

    The role of information processing deficits in poor readers of nonalphabetic scripts such as Chinese is not well documented. Here, we examined perceptual processing in good and poor readers of Chinese. Specifically, two groups of third grade children comprising 20 "good readers" and 19 "poor readers," drawn from a larger pool of 254 students, were…

  12. Aqueous-biamphiphilic ionic liquid systems: self-assembly and synthesis of gold nanocrystals/microplates.

    PubMed

    Rao, K Srinivasa; Trivedi, Tushar J; Kumar, Arvind

    2012-12-13

    Biamphiphilic ionic liquids (BAILs) based on 1,3-dialkylimidazolium cation and alkyl sulfate anions ([C(n)H(2n+1)mim][C(m)H(2m+1)OSO(3)]; n = 4, 6, or 8; m = 8, 12) have been synthesized and characterized for their self-assembling behavior in the aqueous medium. Effects of alteration of alkyl chain length in cation and anion on surfactant properties of BAILs have been examined from surface tension measurements. The effectiveness of surface tension reduction for BAILs has been found to be exceptionally higher as compared to single chain surface active ILs/conventional surfactants. The thermodynamics of the aggregation process has been studied using isothermal titration calorimetry (ITC) and temperature dependent conductivity experiments. Dynamic light scattering (DLS), nuclear magnetic resonance (NMR), and transmission electron microscopy (TEM) studies showed that BAILs formed distinct aggregated structures depending upon the amphiphilic character present in the cation and anion. BAILs ([C(n)H(2n+1)mim][C(m)H(2m+1)OSO(3)]) form micelles when n = 4, 6; m = 8, intermicellar aggregates when n = 4, 6; m = 12, and vesicles when n = 8; m = 8, 12. Gold nanoparticles and microplates have been synthesized in micellar and vesicle solutions of BAILs using a simple photoreduction method. The studies show the potential of BAILs for constructing micelles and supramolecular assemblies, such as bilayer vesicles, which are effective in preparation of nanomaterials of controlled size and morphology.

  13. Antimycobacterial screening of traditional medicinal plants using the microplate resazurin assay.

    PubMed

    Webster, Duncan; Lee, Timothy D G; Moore, Jill; Manning, Tracy; Kunimoto, Dennis; LeBlanc, Darren; Johnson, John A; Gray, Christopher A

    2010-06-01

    Multidrug-resistant Mycobacterium tuberculosis strains have rapidly become a global health concern. North American First Nations communities have used traditional medicines for generations to treat many pulmonary infections. In this study, we evaluated the antimycobacterial activity of 5 medicinal plants traditionally used as general therapeutics for pulmonary illnesses and specifically as treatments for tuberculosis. Aqueous extracts of Aralia nudicaulis, Symplocarpus foetidus, Heracleum maximum, Juniperus communis, and Acorus calamus were screened for antimycobacterial activity against Bacillus Calmette-Guérin, Mycobacterium avium, and M. tuberculosis H37Ra using the colorimetric microplate resazurin assay. Extracts of Acorus calamus and H. maximum root demonstrated significant antimycobacterial activity comparable to that of the rifampin control (2 microg/mL). Evaluation of the cytotoxicity of these 2 extracts using the MTT assay also showed that the extracts were less toxic to 3 human cell lines than was the DMSO positive control. This study demonstrates that aqueous extracts of the roots of H. maximum and Acorus calamus possess strong in vitro antimycobacterial activity, validates traditional knowledge, and provides potential for the development of urgently needed novel antituberculous therapeutics.

  14. Computational modeling and experimental validation of odor detection behaviors of classically conditioned parasitic wasp, Microplitis croceipes.

    PubMed

    Zhou, Zhongkun; Kulasiri, Don; Samarasinghe, Sandhya; Rains, Glen; Olson, Dawn M

    2015-01-01

    A prototype chemical sensor named Wasp hound® that utilizes five classically conditioned parasitoid wasps, Microplitis croceipes (Cresson) (Hymenoptera: Braconidae), to detect volatile odors was successfully implemented in a previous study. To improve the odor-detecting ability of Wasp Hound®, searching behaviors of an individual wasp in a confined area are studied and modeled through stochastic differential equations in this paper. The wasps are conditioned to 20 mg of coffee when associated with food and subsequently, tested to 5, 10, 20, and 40 mg of coffee. A stochastic model is developed and validated based on three positive behavioral responses (walking, rotation around odor source, and self-rotation) from conditioned wasps at four different test dosages. The model is capable to reproducing the behaviors of conditioned wasps, and can be used to improve the ability of Wasp Hound® to assess changes in odor concentration. The model simulation results show the behaviors of conditioned wasps are significantly different when tested at different coffee dosages. We conjecture that the searching behaviors of conditioned wasps are based on the temporal and spatial neuron activity of olfactory receptor neurons and glomeruli, which are strongly correlated to the training dosages. The overall results demonstrate the utility of mathematical models for interpreting experimental observations, gaining novel insights into the dynamic behavior of classically conditioned wasps, as well as broadening the practical uses of Wasp Hound.

  15. Quantitative microplate assay for studying mesenchymal stromal cell-induced neuropoiesis.

    PubMed

    Aizman, Irina; McGrogan, Michael; Case, Casey C

    2013-03-01

    Transplanting mesenchymal stromal cells (MSCs) or their derivatives in a neurodegenerative environment is believed to be beneficial because of the trophic support, migratory guidance, and neurogenic stimuli they provide. There is a growing need for in vitro models of mesenchymal-neural cell interactions to enable identification of mediators of the MSC activity and quantitative assessment of neuropoietic potency of MSC preparations. Here, we characterize a microplate-format coculture system in which primary embryonic rat cortex cells are directly cocultured with human MSCs on cell-derived extracellular matrix (ECM) in the absence of exogenous growth factors. In this system, expression levels of the rat neural stem/early progenitor marker nestin, as well as neuronal and astrocytic markers, directly depended on MSC dose, whereas an oligodendrogenic marker exhibited a biphasic MSC-dose response, as measured using species-specific quantitative reverse transcription-polymerase chain reaction in total cell lysates and confirmed using immunostaining. Both neural cell proliferation and differentiation contributed to the MSC-mediated neuropoiesis. ECM's heparan sulfate proteoglycans were essential for the growth of the nestin-positive cell population. Neutralization studies showed that MSC-derived fibroblast growth factor 2 was a major and diffusible inducer of rat nestin, whereas MSC-derived bone morphogenetic proteins (BMPs), particularly, BMP4, were astrogenesis mediators, predominantly acting in a coculture setting. This system enables analysis of multifactorial MSC-neural cell interactions and can be used for elucidating the neuropoietic potency of MSCs and their derivative preparations.

  16. Geosmin induces genomic instability in the mammalian cell microplate-based comet assay.

    PubMed

    Silva, Aline Flor; Lehmann, Mauricio; Dihl, Rafael Rodrigues

    2015-11-01

    Geosmin (GEO) (trans-1,10-dimethyl-trans-9-decalol) is a metabolite that renders earthy and musty taste and odor to water. Data of GEO genotoxicity on mammalian cells are scarce in the literature. Thus, the present study assessed the genotoxicity of GEO on Chinese hamster ovary (CHO) cells in the microplate-based comet assay. The percent of tail DNA (tail intensity (TI)), tail moment (TM), and tail length (TL) were used as parameters for DNA damage assessment. The results demonstrated that concentrations of GEO of 30 and 60 μg/mL were genotoxic to CHO cells after 4- and 24-h exposure periods, in all parameters evaluated, such as TI, TM, and TL. Additionally, GEO 15 μg/mL was genotoxic in the three parameters only in the 24-h exposure time. The same was observed for GEO 7.5 μg/mL, which induced significant DNA damage observed as TI in the 24-h treatment. The results present evidence that exposure to GEO may be associated with genomic instability in mammalian cells.

  17. Analysis of Genetic Variation across the Encapsidated Genome of Microplitis demolitor Bracovirus in Parasitoid Wasps

    PubMed Central

    Burke, Gaelen R.

    2016-01-01

    Insect parasitoids must complete part of their life cycle within or on another insect, ultimately resulting in the death of the host insect. One group of parasitoid wasps, the ‘microgastroid complex’ (Hymenoptera: Braconidae), engage in an association with beneficial symbiotic viruses that are essential for successful parasitism of hosts. These viruses, known as Bracoviruses, persist in an integrated form in the wasp genome, and activate to replicate in wasp ovaries during development to ultimately be delivered into host insects during parasitism. The lethal nature of host-parasitoid interactions, combined with the involvement of viruses in mediating these interactions, has led to the hypothesis that Bracoviruses are engaged in an arms race with hosts, resulting in recurrent adaptation in viral (and host) genes. Deep sequencing was employed to characterize sequence variation across the encapsidated Bracovirus genome within laboratory and field populations of the parasitoid wasp species Microplitis demolitor. Contrary to expectations, there was a paucity of evidence for positive directional selection among virulence genes, which generally exhibited signatures of purifying selection. These data suggest that the dynamics of host-parasite interactions may not result in recurrent rounds of adaptation, and that adaptation may be more variable in time than previously expected. PMID:27390861

  18. Microplate assay for quantitation of neutral lipids in extracts from microalgae.

    PubMed

    Higgins, Brendan T; Thornton-Dunwoody, Alexander; Labavitch, John M; VanderGheynst, Jean S

    2014-11-15

    Lipid quantitation is widespread in the algae literature, but popular methods such as gravimetry, gas chromatography and mass spectrometry (GC-MS), and Nile red cell staining suffer drawbacks, including poor quantitation of neutral lipids, expensive equipment, and variable results among algae species, respectively. A high-throughput microplate assay was developed that uses Nile red dye to quantify neutral lipids that have been extracted from algae cells. Because the algal extracts contained pigments that quenched Nile red fluorescence, a mild bleach solution was used to destroy pigments, resulting in a nearly linear response for lipid quantities in the range of 0.75 to 40 μg. Corn oil was used as a standard for quantitation, although other vegetable oils displayed a similar response. The assay was tested on lipids extracted from three species of Chlorella and resulted in close agreement with triacylglycerol (TAG) levels determined by thin layer chromatography. The assay was found to more accurately measure algal lipids conducive to biodiesel production and nutrition applications than the widely used gravimetric assay. Assay response was also consistent among different species, in contrast to Nile red cell staining procedures.

  19. Evidence for Moho-lower crustal transition depth diking and rifting of the Sierra Nevada microplate

    NASA Astrophysics Data System (ADS)

    Smith, Kenneth D.; Kent, Graham M.; Seggern, David P.; Driscoll, Neal W.; Eisses, Amy

    2016-10-01

    Lithospheric rifting most often initiates in continental extensional settings where "breaking of a plate" may or may not progress to sea floor spreading. Generally, the strength of the lithosphere is greater than the tectonic forces required for rupture (i.e., the "tectonic force paradox"), and it has been proposed that rifting requires basaltic magmatism (e.g., dike emplacement) to reduce the strength and cause failure, except for the case of a thin lithosphere (<30 km thick). Here we isolate two very similar and unprecedented observations of Moho-lower crustal transition dike or fluid injection earthquake swarms under southern Sierra Valley (SV: 2011-2012) and North Lake Tahoe (LT: 2003-2004), California. These planar distributions of seismicity can be interpreted to define the end points, and cover 25% of the length, of an implied 56 km long structure, each striking N45°W and dipping 50°NE. A single event at 30 km depth that locates on the implied dipping feature between the two swarms is further evidence for a single Moho-transition depth structure. We propose that basaltic or fluid emplacement at or near Moho depths weakens the upper mantle lid, facilitating lithospheric rupture of the Sierra Microplate. Similar to the LT sequence, the SV event is also associated with increased upper crustal seismicity. An 27 October 2011, Mw 4.7 earthquake occurred directly above the deep SV sequence at the base of the upper crustal seismogenic zone ( 15 km depth).

  20. Microplate assay for screening the antibacterial activity of Schiff bases derived from substituted benzopyran-4-one.

    PubMed

    Amin, Rehab M; Abdel-Kader, Nora S; El-Ansary, Aida L

    2012-09-01

    Schiff bases (SB(1)-SB(3)) were synthesized from the condensation of 6-formyl-7-hydroxy-5-methoxy-2-methylbenzopyran-4-one with 2-aminopyridine (SB(1)), p-phenylenediamine (SB(2)) and o-phenylenediamine (SB(3)), while Schiff bases (SB(4)-SB(6)) were synthesized by condensation of 5,7-dihydroxy-6-formyl-2-methylbenzopyran-4-one with 2-aminopyridine (SB(4)), p-phenylenediamine (SB(5)) and o-phenylenediamine (SB(6)). Schiff bases were characterized using elemental analysis, IR, UV-Vis, (1)H NMR, (13)C NMR and mass spectroscopy. These compounds were screened for antibacterial activities by micro-plate assay technique. Escherichia coli and Staphylococcus capitis were exposed to different concentrations of the Schiff bases. Results showed that the antibacterial effect of these Schiff bases on Gram-negative bacteria were higher than that on Gram-positive bacteria moreover, the Schiff bases containing substituent OCH(3) on position five have higher antibacterial activity than that containing hydroxy group on the same position.

  1. The Suckling Hills Fault, Kayak Island Zone, and accretion of the Yakutat microplate, Alaska

    NASA Astrophysics Data System (ADS)

    Chapman, James B.; Worthington, Lindsay L.; Pavlis, Terry L.; Bruhn, Ronald L.; Gulick, Sean P.

    2011-12-01

    The Suckling Hills and Kayak Island are isolated mountain blocks located along strike from each other within the foreland of the St. Elias orogen in southern Alaska. These blocks preserve an erosional surface that was deformed by slip on northwest-dipping reverse faults in the Pleistocene. We suggest that the Suckling Hills Fault and Kayak Island Zone form a segmented fault network that links with the Bering Glacier structure to the north. This fault network separates the central Yakataga fold and thrust belt from complex, multiply deformed structures in the western syntaxis. Ongoing accretion of the Yakutat microplate to North America results in translation of structures of the fold and thrust belt into the western syntaxis. The composite Suckling Hills Fault, Kayak Island Zone, and Bering Glacier structure may have formed because the older structures of the fold and thrust belt were unfavorably oriented within the western syntaxis region. This pattern of deformation provides a template for understanding the complex deformation within the core of the western syntaxis and predicts refolding and straightening of the western syntaxis margin with continued accretion. This study provides an analog for structural overprinting and changing deformation patterns through time in orogenic corners.

  2. Investigation of mitochondrial dysfunction by sequential microplate-based respiration measurements from intact and permeabilized neurons.

    PubMed

    Clerc, Pascaline; Polster, Brian M

    2012-01-01

    Mitochondrial dysfunction is a component of many neurodegenerative conditions. Measurement of oxygen consumption from intact neurons enables evaluation of mitochondrial bioenergetics under conditions that are more physiologically realistic compared to isolated mitochondria. However, mechanistic analysis of mitochondrial function in cells is complicated by changing energy demands and lack of substrate control. Here we describe a technique for sequentially measuring respiration from intact and saponin-permeabilized cortical neurons on single microplates. This technique allows control of substrates to individual electron transport chain complexes following permeabilization, as well as side-by-side comparisons to intact cells. To illustrate the utility of the technique, we demonstrate that inhibition of respiration by the drug KB-R7943 in intact neurons is relieved by delivery of the complex II substrate succinate, but not by complex I substrates, via acute saponin permeabilization. In contrast, methyl succinate, a putative cell permeable complex II substrate, failed to rescue respiration in intact neurons and was a poor complex II substrate in permeabilized cells. Sequential measurements of intact and permeabilized cell respiration should be particularly useful for evaluating indirect mitochondrial toxicity due to drugs or cellular signaling events which cannot be readily studied using isolated mitochondria.

  3. Antibacterial properties of cyclodextrin-antiseptics-complexes determined by microplate laser nephelometry and ATP bioluminescence assay.

    PubMed

    Finger, Susanne; Wiegand, Cornelia; Buschmann, Hans-Jürgen; Hipler, Uta-Christina

    2013-08-16

    Cyclodextrins (CDs) are able to form inclusion complexes with other molecules, thereby, protecting these guest molecules from degradation, enhancing their biocompatibility or influencing their physiological distribution while retaining their activity. Here, antibacterial effects of CD-complexes with the antiseptics chlorhexidine diacetate (CHX), iodine (IOD) and polihexanide (PHMB) were determined using two different in vitro methods, microplate laser nephelometry and an ATP bioluminescence assay. Laser nephelometry is a direct method for monitoring and evaluating growth of micro-organisms by measurement of the turbidity of the solution. In contrast, the ATP bioluminescence assay determines specifically the amount of metabolic active bacterial cells. The antibacterial effects of CD-antiseptics-complexes were examined for Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus and Staphylococcus epidermidis and the results of both methods were compared in respect of calculated means of half maximal inhibitory concentrations (IC50) and statistical evaluated Pearson's correlation coefficients (r). It could be demonstrated that both methods showed a high comparability although they differ in the parameters tested. This study revealed that CD-complexes with CHX and PHMB were most effective against E. coli and the tested staphylococci. While CD-IOD-complexes obtained high activity against K. pneumoniae, P. aeruginosa was distinctly more resistant compared to the other bacteria.

  4. Passive microfluidic array card and reader

    DOEpatents

    Dugan, Lawrence Christopher; Coleman, Matthew A.

    2011-08-09

    A microfluidic array card and reader system for analyzing a sample. The microfluidic array card includes a sample loading section for loading the sample onto the microfluidic array card, a multiplicity of array windows, and a transport section or sections for transporting the sample from the sample loading section to the array windows. The microfluidic array card reader includes a housing, a receiving section for receiving the microfluidic array card, a viewing section, and a light source that directs light to the array window of the microfluidic array card and to the viewing section.

  5. Towards a Culturally Situated Reader Response Theory

    ERIC Educational Resources Information Center

    Brooks, Wanda; Browne, Susan

    2012-01-01

    This article describes a theory of how culture enables literary interpretations of texts. We begin with a brief overview of the reader response field. From there, we introduce the theory and provide illustrative participant data examples. These data examples illustrate the four cultural positions middle grade students in our research assumed when…

  6. Reflections on Teaching Struggling Middle School Readers.

    ERIC Educational Resources Information Center

    Ivey, Gay

    1999-01-01

    Shares four working generalizations on what it takes for middle school students with persistent reading difficulties to become successful readers: (1) access to materials that span the gamut of interests and difficulty levels; (2) opportunities to share reading experiences with teachers and classmates; (3) real purposes for reading; and (4)…

  7. The Copenhagen Studies in Reader Response.

    ERIC Educational Resources Information Center

    Dollerup, Cay; And Others

    1990-01-01

    This article describes a series of Scandinavian studies in reader response from 1968 to 1990. Studies chronologically discussed in the article are: (1) "Rhythm in Poetry"; (2) "The Esthetic Experience"; (3) "Meaning in Literary Texts"; (4) "Tension"; (5) the "Ram" study; (6) the…

  8. Project Physics Reader 4, Light and Electromagnetism.

    ERIC Educational Resources Information Center

    Harvard Univ., Cambridge, MA. Harvard Project Physics.

    As a supplement to Project Physics Unit 4, a collection of articles is presented in this reader for student browsing. The 21 articles are included under the following headings: Letter from Thomas Jefferson; On the Method of Theoretical Physics; Systems, Feedback, Cybernetics; Velocity of Light; Popular Applications of Polarized Light; Eye and…

  9. Getting Ready to Read with Readers Theatre

    ERIC Educational Resources Information Center

    Barchers, Suzanne I; Pfeffinger, Charla R.

    2007-01-01

    Readers theatre is a presentation by two or more participants who read from scripts and interpret a literary work in such a way that the audience imaginatively senses characterization, setting, and action. This book offers 50, two-page reproducible scripts to entice the preschool and kindergarten group into beginning to read. These patterned…

  10. The Expectant Reader in Theory and Practice.

    ERIC Educational Resources Information Center

    Fowler, Lois Josephs; McCormick, Kathleen

    1986-01-01

    Offers a method of using reader response theory that emphasizes the expectations about a text and how those expectations are fulfilled or deflated. Specifically, students read traditional fables, fairy tales, and parables, and compare them to contemporary works such as Kafka's "Metamorphosis" and Marquez's "The Very Old Man With Enormous Wings."…

  11. Vocabulary Instruction for Second Language Readers

    ERIC Educational Resources Information Center

    Nisbet, Deanna L.

    2010-01-01

    Over the past 20 years, research has consistently affirmed the importance of explicit vocabulary instruction for adult learners of English as a second language (ESL). Given the significant vocabulary demands faced by adult second language readers, ESL teachers must carefully target their instruction for maximum impact and to foster meaningful…

  12. How and Why Stories for Readers Theatre

    ERIC Educational Resources Information Center

    Wolfman, Judy

    2004-01-01

    How did the bee get his bumble? How do birds get their feathers? Why is the bluebird blue? Curious first through fifth graders want to know how and why things happen! Judy Wolfman has created 40 Readers Theatre scripts based on imaginative and creative porquoi stories that stem from multicultural folktales as well as Native American Indian legends…

  13. Children's Identities as Readers and Writers

    ERIC Educational Resources Information Center

    Braccio, Danielle

    2006-01-01

    In this article, the author shares the reflections of second graders at PS 11, an elementary school in the Chelsea section of Manhattan, on their identities as readers and writers through a photo essay. It is through these two mediums that they are coming to know themselves as learners and as human beings. One example of student comments is, Billy…

  14. Reader Reactions to Color in Newspapers.

    ERIC Educational Resources Information Center

    Bohle, Robert H.; Garcia, Mario R.

    In order to discover reader reactions to color on a newspaper page, specifically eye movement and overall opinion of the paper, identical pages were created and printed by the "St. Petersburg Times" (Florida). The content of fifteen front pages, six lifestyles pages, and three sports front pages were nearly identical, differing only in the kind…

  15. Shakespeare and Reader's Theatre: Fellow Traveling Companions

    ERIC Educational Resources Information Center

    Ratliff, Gerald Lee

    2010-01-01

    Whether constructed on literary analysis models or inspired by conventional acting theories, Reader's Theatre performance techniques are an invaluable instructional tool available to teachers who want their students to see, hear and feel Shakespeare texts in classroom discussion and performance. These exercises are designed to promote both a…

  16. Transcending Bias through Reader-Response Theory

    ERIC Educational Resources Information Center

    Soublis, Theoni; Winkler, Erik

    2004-01-01

    The preservice teachers from all disciplines will be benefited if they incorporate reading in their classes according to Dr. Louise Rosenblatt's reader-response theory. A teacher's experience with her students while reading Chris Crutcher's "Staying Fat for Sarah Byanes" in the Secondary Content Area and a student's response on the novel are…

  17. Effective Instruction for Struggling Readers, K-6

    ERIC Educational Resources Information Center

    Taylor, Barbara M., Ed.; Ysseldyke, James, E., Ed.

    2007-01-01

    With contributions from recognized leaders in the field of reading interventions, this book examines what has been learned from research to help struggling readers in grades K-6. Focusing on strategies that have been proven effective, the authors help educators meet the demands placed on them to ensure that all students are making good progress…

  18. Readers Response Approach to English Poetry Teaching.

    ERIC Educational Resources Information Center

    Lin, Antonia Hsiu-Chen; Sher, Teresa Hsiang-Jen

    This paper describes an elective course at Taiwan's Wen Tzao Ursuline College of Modern Languages, "Concise English Poetry Appreciation and Recitation." The course is based on the reader response approach and targets third year students, leading them into the world of poetry through various stages (traditional nursery rhymes and simple,…

  19. Teaching Social Studies Using Basal Readers.

    ERIC Educational Resources Information Center

    Garcia, Jesus; Logan, John W.

    1983-01-01

    A lesson, "Harriet Tubman: A Most Successful Conductor," illustrates how to employ a basal reader in social studies instruction in the elementary grades. This approach offers students a relevant curriculum, greater opportunities for concept development, practice in skills areas, and activities that offer greater opportunity to master…

  20. Reading Strategies for Beginning and Proficient Readers.

    ERIC Educational Resources Information Center

    Atterman, Jennifer S.

    The single most important task facing elementary school teachers today is teaching students to read by the end of third grade. Learning to read in those formative years is essential to develop the higher order thinking skills demanded in the older grades, when students are reading to learn. Beginning readers must be engaged in highly purposeful…

  1. Values in American and Hispanic Children's Readers.

    ERIC Educational Resources Information Center

    Knafle, June D.; And Others

    1991-01-01

    Assessment of the values (positive behavior, positive feelings, negative behavior, negative feelings, traditional values, Judeo-Christian religious values, other religious values, and neutral situations) presented in both American and Hispanic basal reader series found several differences among publishers and cultural groups that lead to…

  2. Oral Interpretation of Literature: Readers' Theater

    ERIC Educational Resources Information Center

    Kennedy, Joan

    2011-01-01

    The pedagogical principle of experiential learning embodied in the oral interpretation of literature through Readers' Theater provides an avenue to accomplish a seemingly daunting task. Students' participation in reading, interpreting, discussing, writing, assessing, and performing their own creative responses to a literary work promotes a…

  3. eReaders in School Libraries

    ERIC Educational Resources Information Center

    Sutton, Amy

    2013-01-01

    This article explores the possibility of using eReaders in the schools of the Houston Independent School District (HISD), specifically the Barnes and Noble Nook Pilot Project. HSID initially approved only an eReading device that was not wireless since it is very strict on the devices granted access to the wireless network. The biggest roadblock…

  4. Volunteer Expert Readers for STEM Student Writers

    ERIC Educational Resources Information Center

    Moskovitz, Cary

    2014-01-01

    This paper reports on a novel approach to providing undergraduates with feedback on STEM writing assignments via an otherwise untapped educational resource: university alumni and employees who normally play no role in the institution's educational mission. In the Volunteer Expert Reader (VER) approach, students are paired with volunteers whose…

  5. Project Physics Reader 6, The Nucleus.

    ERIC Educational Resources Information Center

    Harvard Univ., Cambridge, MA. Harvard Project Physics.

    As a supplement to Project Physics Unit 6, a collection of articles is presented in this reader for student browsing. Five excerpts are concerned with the nuclear energy revolution, the 20th birthday and possible consequences of the atomic age, a scientist's view of science, and relations between mathematics and physics. Six book passages are…

  6. Washington Striving Readers: Year 1 Evaluation Report

    ERIC Educational Resources Information Center

    Deussen, Theresa; Scott, Caitlin; Nelsestuen, Kari; Roccograndi, Angela; Davis, Ann

    2012-01-01

    In 2009, the United States Department of Education conducted a competition for a second round of Striving Readers grants. Its dual purpose was to: (1) Raise middle and high school students' literacy levels in Title I-eligible schools with significant numbers of students reading below grade level; and (2) Build a strong, scientific research base…

  7. The Emergent Reader's Working Kit of Stereotypes

    ERIC Educational Resources Information Center

    Mackey, Margaret

    2013-01-01

    This article draws on a careful study of series fiction read in the 1950s to explore how stereotypes feature in the development of a young reader's competence in learning to process stories in print. Five categories of stereotype are teased out: "embodied stereotypes," understood through physical experience; "working stereotypes," discerned…

  8. Japanese-American Internment. A Historical Reader.

    ERIC Educational Resources Information Center

    2000

    This Historical Reader on "Japanese American Internment" in World War II introduces students to key events and issues during the period through the voices of people with firsthand experienced. Source documents and illustrations are arranged in chronological order and/or thematic units that establish context. Each selection is followed by…

  9. Print Readers' Perceptions of Various Advertising Formats.

    ERIC Educational Resources Information Center

    Lamb, Charles W., Jr.; And Others

    1979-01-01

    The findings of a study of print readers' perceptions of the believability and interest of various advertising formats fail to support increasing either the frequency or specificity of comparative messages. The findings suggest that advertisers should consider the dimensions of intensity and directionality in their message development. (GT)

  10. Teaching Content Material through Reader's Theater

    ERIC Educational Resources Information Center

    Forney, Melissa

    2013-01-01

    When it comes to content area material, much of what students read and learn is predicated on information they have read before and are supposed to remember. Teachers often use silent reading and round robin reading as preferred reading methods to help students learn content area material. The objective of this study was to test reader's theater…

  11. The Author as Reader and Writer

    ERIC Educational Resources Information Center

    Miller, Tom

    2012-01-01

    This article presents contemporary commentary on the previously published articles "Writing for the Reader: A Problem-Solution Approach" and "Motivating Learners at South Korean Universities." Having been out of the field of English as a foreign language for several years, the author was surprised and pleased when he was asked to write some…

  12. Reader Response: Young Children Can Do That!

    ERIC Educational Resources Information Center

    Lowe, Kaye; Johnston, Cammie

    2000-01-01

    Describes "reader response" teaching techniques to enhance young children's critical thinking skills, build a sound literacy foundation, and clarify the relationship between reading and writing. Provides examples of response logs/journal entries, aesthetic responses such as drawings and dramatic reenactments, and student discussions…

  13. Character and Moral Education: A Reader

    ERIC Educational Resources Information Center

    DeVitis, Joseph L., Ed.; Yu, Tianlong, Ed.

    2011-01-01

    Against a formidable national discourse that emphasizes academic standardization, accountability, and high-stakes testing in educational policy, "Character and Moral Education: A Reader" seeks to re-introduce and revive the moral mission of education in public conversation and practices in America's schools. With contributions from a…

  14. KURDISH READERS. PART I, NEWSPAPER KURDISH.

    ERIC Educational Resources Information Center

    ABDULLA, JAMAL JALAL; MCCARUS, ERNEST N.

    ASSUMING A MASTERY OF THE CONTENTS OF THE "BASIC COURSE IN KURDISH" (BY THE SAME AUTHORS), THIS READER PRESENTS A VARIETY OF 28 ARTICLES SELECTED FROM THE IRAQI NEWSPAPERS "ZHIN" AND "KHEBAT." EACH LESSON BEGINS WITH A SELECTION WRITTEN IN KURDISH (MODIFIED ARABIC-PERSIAN) SCRIPT, FOLLOWED BY PHONEMIC TRANSCRIPTION…

  15. 38 CFR 21.150 - Reader service.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Special Rehabilitation Services § 21.150 Reader service. (a) Limitations on vision. A veteran considered... vision is 20/200 in both eyes; (2) Whose central vision is greater than 20/200 but whose field of vision... greater than 20 degrees; or (3) With impaired vision, whose condition or prognosis indicates that...

  16. 38 CFR 21.150 - Reader service.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Special Rehabilitation Services § 21.150 Reader service. (a) Limitations on vision. A veteran considered... vision is 20/200 in both eyes; (2) Whose central vision is greater than 20/200 but whose field of vision... greater than 20 degrees; or (3) With impaired vision, whose condition or prognosis indicates that...

  17. Classic Readers Theatre for Young Adults.

    ERIC Educational Resources Information Center

    Barchers, Suzanne I.; Kroll, Jennifer L.

    This book presents 16 original scripts that have been adapted from classic works of literature for use for readers theatre with young adults and ESL (English as a Second Language) students. Adaptations of the following works are included: "Little Women" (Louisa May Alcott); episodes from "Don Quixote" (Miguel de Cervantes; "The Necklace" (Guy de…

  18. Tropospheric Emission Spectrometer Product File Readers

    NASA Technical Reports Server (NTRS)

    Fisher, Brendan M.

    2010-01-01

    TES Product File Reader software extracts data from publicly available Tropospheric Emission Spectrometer (TES) HDF (Hierarchical Data Format) product data files using publicly available format specifications for scientific analysis in IDL (interactive data language). In this innovation, the software returns data fields as simple arrays for a given file. A file name is provided, and the contents are returned as simple IDL variables.

  19. Critical Literacy as Comprehension: Expanding Reader Response

    ERIC Educational Resources Information Center

    McLaughlin, Maureen; De Voogd, Glenn

    2004-01-01

    This article presents the theoretical underpinnings of critical literacy and related principles. It also provides ideas for creating environments to promote reading from a critical stance, teaching strategies, sample classroom applications, and annotated lists of theme-related texts. When engaging in critical literacy, readers move beyond…

  20. Reader-Response and the Pathos Principle.

    ERIC Educational Resources Information Center

    Johnson, Nan

    1988-01-01

    Reviews and equates theories of reader-response and rhetorical theories on audience response (the pathos principle). Concludes that the fundamental synonymity between them represents a significant bridge between analysis of literary texts and the dynamics of formal and social discourse and provides a theoretical foundation for teaching reading and…

  1. Trade Related Reading Packets for Disabled Readers.

    ERIC Educational Resources Information Center

    Davis, Beverly; Woodruff, Nancy S.

    Six trade-related reading packets for disabled readers are provided for these trades: assemblers, baking, building maintenance, data entry, interior landscaping, and warehousing. Each packet stresses from 9 to 14 skills. Those skills common to most packets include context clues, fact or opinion, details, following directions, main idea,…

  2. Readerly and Writerly "Letters from the Park."

    ERIC Educational Resources Information Center

    Conde, Susana

    1993-01-01

    Discusses in depth the film "Cartas del parque" ("Letters from the Park"), the first of six films in the "Amores Dificiles" series. Notes that the film is pervaded by the traditional overdetermination of gender roles. Suggests that an intrusive and authoritative narrator makes of this both a "readerly" and a…

  3. Understanding Deaf Readers: An Interpretative Phenomenological Analysis

    ERIC Educational Resources Information Center

    Kelstone, Aaron Weir

    2013-01-01

    The development of reading skills, beyond a functional level, is difficult for most deaf readers. Standardized testing demonstrates a median 4th grade reading level that remains consistent even after national norming of the Stanford Achievement test on the population of deaf school children. Deaf education continues to generate various educational…

  4. Break-up of Pangaea and Tectonic History of the Adria Microplate

    NASA Astrophysics Data System (ADS)

    Schettino, A.; Turco, E.

    2008-12-01

    A new kinematic model is proposed for the tectonic evolution of the western Tethys and the Adria microplate, constrained by the Atlantic plate kinematics and on land geologic evidences. The model defines thirteen tectonic phases, spanning the time interval from the late Ladinian (230 Ma) to the present. During the first phase, from the late Ladinian (230 Ma) to the latest Rhaetian (200 Ma), rifting proceeded along the eastern margin of North America, the northwest African margin, the High, Saharan and Tunisian Atlas, determining the formation of a separate Moroccan microplate at the interface between Gondwana and Laurasia. To the East, the break-up of Pangaea proceeded through a new phase of spreading in the central Mediterranean (Lagonegro basin) and the Tethys Ocean, while Adria remained attached to the southern Eurasian margin. During the second phase, from the latest Rhaetian (200 Ma) to the late Pliensbachian (185 Ma), oceanic crust started forming between the East Coast and Blake Spur magnetic anomalies, while the Morrocan Meseta simply continued to rift away from North America. During this time interval the Atlas rift reached its maximum extent. In the western Tethys region, cessation of spreading in the Lagonegro and Ionian Basins was accompanied by the formation of a new plate boundary which rifted Adria from Eurasia. The third phase, from the late Pliensbachian (185 Ma) to chron M21 (147.7 Ma), was triggered by the northward jump of the main plate boundary connecting the central Atlantic with the Tethys area. Therefore, as soon as rifting in the Atlas zone ceased, plate motion started along complex fault systems between Morocco and Iberia, while a rift/drift transition occurred in the northern segment of the central Atlantic, between Morocco and the conjugate margin of Nova Scotia. Sea floor spreading also started in the Mediterranean area, forming the Ligurian and Alpine Tethys oceans. The next two phases, from chron M21 (147.7 Ma) to chron M0 (120

  5. Quantitative Microplate-Based Growth Assay for Determination of Antifungal Susceptibility of Histoplasma capsulatum Yeasts

    PubMed Central

    Goughenour, Kristie D.; Balada-Llasat, Joan-Miquel

    2015-01-01

    Standardized methodologies for determining the antifungal susceptibility of fungal pathogens is central to the clinical management of invasive fungal disease. Yeast-form fungi can be tested using broth macrodilution and microdilution assays. Reference procedures exist for Candida species and Cryptococcus yeasts; however, no standardized methods have been developed for testing the antifungal susceptibility of yeast forms of the dimorphic systemic fungal pathogens. For the dimorphic fungal pathogen Histoplasma capsulatum, susceptibility to echinocandins differs for the yeast and the filamentous forms, which highlights the need to employ Histoplasma yeasts, not hyphae, in antifungal susceptibility tests. To address this, we developed and optimized methodology for the 96-well microtiter plate-based measurement of Histoplasma yeast growth in vitro. Using optical density, the assay is quantitative for fungal growth with a dynamic range greater than 30-fold. Concentration and assay reaction time parameters were also optimized for colorimetric (MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] reduction) and fluorescent (resazurin reduction) indicators of fungal vitality. We employed this microtiter-based assay to determine the antifungal susceptibility patterns of multiple clinical isolates of Histoplasma representing different phylogenetic groups. This methodology fulfills a critical need for the ability to monitor the effectiveness of antifungals on Histoplasma yeasts, the morphological form present in mammalian hosts and, thus, the form most relevant to disease. PMID:26246483

  6. Immunocapture and microplate-based activity measurement of mammalian pyruvate dehydrogenase complex.

    PubMed

    Lib, Margarita; Rodriguez-Mari, Adriana; Marusich, Michael F; Capaldi, Roderick A

    2003-03-01

    Altered pyruvate dehydrogenase (PDH) functioning occurs in primary PDH deficiencies and in diabetes, starvation, sepsis, and possibly Alzheimer's disease. Currently, the activity of the enzyme complex is difficult to measure in a rapid high-throughput format. Here we describe the use of a monoclonal antibody raised against the E2 subunit to immunocapture the intact PDH complex still active when bound to 96-well plates. Enzyme turnover was measured by following NADH production spectrophotometrically or by a fluorescence assay on mitochondrial protein preparations in the range of 0.4 to 5.0 micro g per well. Activity is sensitive to known PDH inhibitors and remains regulated by phosphorylation and dephosphorylation after immunopurification because of the presence of bound PDH kinase(s) and phosphatase(s). It is shown that the immunocapture assay can be used to detect PDH deficiency in cell extracts of cultured fibroblasts from patients, making it useful in patient screens, as well as in the high-throughput format for discovery of new modulators of PDH functioning.

  7. Electrophysiological and behavioral responses of Microplitis mediator (Hymenoptera: Braconidae) to caterpillar-induced volatiles from cotton.

    PubMed

    Yu, Huilin; Zhang, Yongjun; Wyckhuys, Kris A G; Wu, Kongming; Gao, Xiwu; Guo, Yuyuan

    2010-04-01

    Microplitis mediator Haliday (Hymenoptera: Braconidae) is an important larval endoparasitoid of various lepidopteran pests, including Helicoverpa armigera (Hübner). In China, H. armigera is a key pest of cotton and is currently the focus of several biological control efforts that use M. mediator as principal natural enemy of this pest. To improve the success of biological control efforts, behavioral studies are needed that shed light on the interaction between M. mediator and H. armigera. In this study, we determined M. mediator response to volatile compounds from undamaged, mechanically injured, or H. armigera--damaged plants and identified attractive volatiles. In Y-tube olfactometer assays, we found that mechanically damaged plants and/or plants treated with H. armigera oral secretions did not attract wasps. However, volatiles from H. armigera-damaged plants elicited a strong attraction of both M. mediator sexes. Headspace extracts from H. armigera-damaged cotton were analyzed by coupled gas chromatography-electroantennographic detection (GC-EAD), and a total of seven different compounds were found to elicit electroantennogram (EAG) responses, including an unknown compound. Six different EAD-active volatiles were identified from caterpillar-damaged cotton plants, of which 3, 7-dimethyl-1, 3, 6-octatriene and (Z)-3-hexenyl acetate were the principal compounds. Olfactometer assays indicated that individual synthetic compounds of 3, 7-dimethyl-1, 3, 6-octatriene, (Z)-3-hexenyl acetate, and nonanal were attractive to M. mediator. Field cage studies showed that parasitism of H. armigera larvae by M. mediator was higher on cotton plants to which 3,7-dimethyl-1,3, 6-octatriene was applied. Our results show that the combination of terpenoids and green leaf volatiles may not only facilitate host, mate, or food location but may also increase H. armigera parasitism by M. mediator.

  8. Microplitis demolitor bracovirus genome segments vary in abundance and are individually packaged in virions

    SciTech Connect

    Beck, Markus H.; Inman, Ross B.; Strand, Michael R. . E-mail: mrstrand@bugs.ent.uga.edu

    2007-03-01

    Polydnaviruses (PDVs) are distinguished by their unique association with parasitoid wasps and their segmented, double-stranded (ds) DNA genomes that are non-equimolar in abundance. Relatively little is actually known, however, about genome packaging or segment abundance of these viruses. Here, we conducted electron microscopy (EM) and real-time polymerase chain reaction (PCR) studies to characterize packaging and segment abundance of Microplitis demolitor bracovirus (MdBV). Like other PDVs, MdBV replicates in the ovaries of females where virions accumulate to form a suspension called calyx fluid. Wasps then inject a quantity of calyx fluid when ovipositing into hosts. The MdBV genome consists of 15 segments that range from 3.6 (segment A) to 34.3 kb (segment O). EM analysis indicated that MdBV virions contain a single nucleocapsid that encapsidates one circular DNA of variable size. We developed a semi-quantitative real-time PCR assay using SYBR Green I. This assay indicated that five (J, O, H, N and B) segments of the MdBV genome accounted for more than 60% of the viral DNAs in calyx fluid. Estimates of relative segment abundance using our real-time PCR assay were also very similar to DNA size distributions determined from micrographs. Analysis of parasitized Pseudoplusia includens larvae indicated that copy number of MdBV segments C, B and J varied between hosts but their relative abundance within a host was virtually identical to their abundance in calyx fluid. Among-tissue assays indicated that each viral segment was most abundant in hemocytes and least abundant in salivary glands. However, the relative abundance of each segment to one another was similar in all tissues. We also found no clear relationship between MdBV segment and transcript abundance in hemocytes and fat body.

  9. Long-term GNSS measurements from the northern Adria microplate reveal fault-induced fluid mobilization

    NASA Astrophysics Data System (ADS)

    Rossi, Giuliana; Zuliani, David; Fabris, Paolo

    2016-10-01

    Due to the development of denser permanent Global Navigation Satellite System (GNSS) networks over the last decade, the observation of transient deformations has significantly increased, mainly in high strain-rate zones. We analyzed the data from a group of permanent GNSS sites on the N-Adria microplate, where anomalous southward tilting and low-frequency tremors preceded the 1976 Mw = 6.4 earthquake. We present records from different stations of a transient signal with an approximately 2-year period that propagated through the northern edge of Adria, in a region 150 km wide. This represents the first time a transient deformation event has been observed in a continental collision area. We exclude surface and groundwater hydrological load effects because we corrected the data for such effects at seasonal, annual, and multiyear scales. The movement is initially upward, except in one location, with slight tilting parallel to the direction of the main tectonic structures. Later, the opposite behavior is observed. The novel methods used include earthquake location techniques and tomographic inversion of the arrival times. These methods demonstrated that the transient source was located spatially and temporally close to the 2004 Mw = 5.2 event in Bovec (Slovenia), attributed to the activity of the Ravne fault. We interpret the transient rises as the expression of a porosity wave, possibly produced by fault valve behavior of the NW tip of the Ravne fault. The propagation velocity is consistent with this hypothesis. As a further test, we invert the arrival times of the transient through hydraulic tomography to obtain hydraulic diffusivity: the values are compatible with the lithotypes present in the region and the literature. By substituting the tomographic velocity and diffusivity in the solitary/porosity wave equation, we infer an initial effective stress of approximately 0.23 bar, sufficient to alter the equilibrium of some fault segments and influence the subsequent

  10. Free vibration and biaxial buckling analysis of magneto-electro-elastic microplate resting on visco-Pasternak substrate via modified strain gradient theory

    NASA Astrophysics Data System (ADS)

    Jamalpoor, A.; Ahmadi-Savadkoohi, A.; Hosseini-Hashemi, Sh

    2016-10-01

    This paper deals with the theoretical analysis of free vibration and biaxial buckling of magneto-electro-elastic (MEE) microplate resting on Kelvin-Voigt visco-Pasternak foundation and subjected to initial external electric and magnetic potentials, using modified strain gradient theory (MSGT). Kirchhoff plate model and Hamilton’s principle are employed to extract the governing equations of motion. Governing equations were analytically solved to obtain clear closed-form expression for complex natural frequencies and buckling loads using Navier’s approach. Numerical results are presented to reveal variations of natural frequency and buckling load ratio of MEE microplate against different amounts of the length scale parameter, initial external electric and magnetic potentials, aspect ratio, damping and transverse and shear stiffness parameters of the visco-Pasternak foundation, length to thickness ratio, microplate thickness and higher modes. Numerical results of this study illustrate that by increasing thickness-to-material length scale parameter ratio, both natural frequency and buckling load ratio predicted by MSGT and modified couple stress theory are reduced because the non-dimensional length scale parameter tends to decrease the stiffness of structures and make them more flexible. In addition, results show that initial external electric and initial external magnetic potentials have no considerable influence on the buckling load ratio and frequency of MEE microplate as the microplate thickness increases.

  11. The Dinaric fault system: Large-scale structure, rates of slip, and Plio-Pleistocene evolution of the transpressive northeastern boundary of the Adria microplate

    NASA Astrophysics Data System (ADS)

    Moulin, Adrien; Benedetti, Lucilla; Rizza, Magali; Jamšek Rupnik, Petra; Gosar, Andrej; Bourlès, Didier; Keddadouche, Karim; Aumaître, Georges; Arnold, Maurice; Guillou, Valery; Ritz, Jean-François

    2016-10-01

    Located at the northeastern corner of the Adria microplate, the Alps-Dinarides junction represents a key region for understanding how the Adria microplate interacts with stable Europe. However, little is known on how the present-day deformation imposed by the rotation of the Adria microplate is absorbed across the Dinarides. Using morphotectonic analysis based on satellite and aerial images, accurate topographical maps, and digital elevation models combined with field investigations, we mapped in detail the three main active faults of the Northern Dinarides. Geomorphic and geological cumulative displacements ranging from a few meters to several kilometers have been identified on those faults and dated for the most recent ones using 36Cl exposure dating. Those results yielded a total right-lateral motion of 3.8 ± 0.7 mm/yr oriented N317. Comparing our results with the motion expected from Adria rotation models suggests that the Northern Dinarides absorbs most of the predicted Adria-Eurasia motion, thus representing the eastern boundary of the microplate. However, a significant E-W component is lacking, suggesting that part of the stress imposed by the microplate rotation is transferred farther to the east. Finally, bounds placed on the Plio-Pleistocene kinematics confirm that faulting onset occurred during the Early Pliocene and evidence a significant kinematic change at the Early/Middle Pleistocene boundary.

  12. Blind Braille readers mislocate tactile stimuli.

    PubMed

    Sterr, Annette; Green, Lisa; Elbert, Thomas

    2003-05-01

    In a previous experiment, we observed that blind Braille readers produce errors when asked to identify on which finger of one hand a light tactile stimulus had occurred. With the present study, we aimed to specify the characteristics of this perceptual error in blind and sighted participants. The experiment confirmed that blind Braille readers mislocalised tactile stimuli more often than sighted controls, and that the localisation errors occurred significantly more often at the right reading hand than at the non-reading hand. Most importantly, we discovered that the reading fingers showed the smallest error frequency, but the highest rate of stimulus attribution. The dissociation of perceiving and locating tactile stimuli in the blind suggests altered tactile information processing. Neuroplasticity, changes in tactile attention mechanisms as well as the idea that blind persons may employ different strategies for tactile exploration and object localisation are discussed as possible explanations for the results obtained.

  13. A Microplate Growth Inhibition Assay for Screening Bacteriocins against Listeria monocytogenes to Differentiate Their Mode-of-Action.

    PubMed

    Vijayakumar, Paul Priyesh; Muriana, Peter M

    2015-06-11

    Lactic acid bacteria (LAB) have historically been used in food fermentations to preserve foods and are generally-recognized-as-safe (GRAS) by the FDA for use as food ingredients. In addition to lactic acid; some strains also produce bacteriocins that have been proposed for use as food preservatives. In this study we examined the inhibition of Listeria monocytogenes 39-2 by neutralized and non-neutralized bacteriocin preparations (Bac+ preps) produced by Lactobacillus curvatus FS47; Lb. curvatus Beef3; Pediococcus acidilactici Bac3; Lactococcus lactis FLS1; Enterococcus faecium FS56-1; and Enterococcus thailandicus FS92. Activity differences between non-neutralized and neutralized Bac+ preps in agar spot assays could not readily be attributed to acid because a bacteriocin-negative control strain was not inhibitory to Listeria in these assays. When neutralized and non-neutralized Bac+ preps were used in microplate growth inhibition assays against L. monocytogenes 39-2 we observed some differences attributed to acid inhibition. A microplate growth inhibition assay was used to compare inhibitory reactions of wild-type and bacteriocin-resistant variants of L. monocytogenes to differentiate bacteriocins with different modes-of-action (MOA) whereby curvaticins FS47 and Beef3, and pediocin Bac3 were categorized to be in MOA1; enterocins FS92 and FS56-1 in MOA2; and lacticin FLS1 in MOA3. The microplate bacteriocin MOA assay establishes a platform to evaluate the best combination of bacteriocin preparations for use in food applications as biopreservatives against L. monocytogenes.

  14. Using Reader Response in a College Literature Class.

    ERIC Educational Resources Information Center

    Soles, Derek

    1995-01-01

    Claims that the insights of reader response theory can be brought into the teaching of poetry in college literature courses. Outlines methods for utilizing reader response techniques to help students enjoy and understand poetry. (HB)

  15. Plastic Logic quits e-reader market

    NASA Astrophysics Data System (ADS)

    Perks, Simon

    2012-07-01

    A UK firm spun out from the University of Cambridge that sought to be a world leader in flexible organic electronic circuits and displays has pulled out of the competitive e-reader market as it struggles to find a commercial outlet for its technology. Plastic Logic announced in May that it is to close its development facility in Mountain View, California, with the loss of around 40 jobs.

  16. Man and His World; A Structured Reader.

    ERIC Educational Resources Information Center

    Kurilecz, Margaret

    This reader for high intermediate or advanced students of English as a second language may be used on the secondary level for students from 14 to 18 years of age who have had about five years of instruction in daily classes, or on the adult level for students who have had two years of work. With a three-way emphasis on reading, speaking, and…

  17. E-Readers and Visual Fatigue

    PubMed Central

    Benedetto, Simone; Drai-Zerbib, Véronique; Pedrotti, Marco; Tissier, Geoffrey; Baccino, Thierry

    2013-01-01

    The mass digitization of books is changing the way information is created, disseminated and displayed. Electronic book readers (e-readers) generally refer to two main display technologies: the electronic ink (E-ink) and the liquid crystal display (LCD). Both technologies have advantages and disadvantages, but the question whether one or the other triggers less visual fatigue is still open. The aim of the present research was to study the effects of the display technology on visual fatigue. To this end, participants performed a longitudinal study in which two last generation e-readers (LCD, E-ink) and paper book were tested in three different prolonged reading sessions separated by - on average - ten days. Results from both objective (Blinks per second) and subjective (Visual Fatigue Scale) measures suggested that reading on the LCD (Kindle Fire HD) triggers higher visual fatigue with respect to both the E-ink (Kindle Paperwhite) and the paper book. The absence of differences between E-ink and paper suggests that, concerning visual fatigue, the E-ink is indeed very similar to the paper. PMID:24386252

  18. E-readers and visual fatigue.

    PubMed

    Benedetto, Simone; Drai-Zerbib, Véronique; Pedrotti, Marco; Tissier, Geoffrey; Baccino, Thierry

    2013-01-01

    The mass digitization of books is changing the way information is created, disseminated and displayed. Electronic book readers (e-readers) generally refer to two main display technologies: the electronic ink (E-ink) and the liquid crystal display (LCD). Both technologies have advantages and disadvantages, but the question whether one or the other triggers less visual fatigue is still open. The aim of the present research was to study the effects of the display technology on visual fatigue. To this end, participants performed a longitudinal study in which two last generation e-readers (LCD, E-ink) and paper book were tested in three different prolonged reading sessions separated by--on average--ten days. Results from both objective (Blinks per second) and subjective (Visual Fatigue Scale) measures suggested that reading on the LCD (Kindle Fire HD) triggers higher visual fatigue with respect to both the E-ink (Kindle Paperwhite) and the paper book. The absence of differences between E-ink and paper suggests that, concerning visual fatigue, the E-ink is indeed very similar to the paper.

  19. Directional Radio-Frequency Identification Tag Reader

    NASA Technical Reports Server (NTRS)

    Medelius, Pedro J.; Taylor, John D.; Henderson, John J.

    2004-01-01

    A directional radio-frequency identification (RFID) tag reader has been designed to facilitate finding a specific object among many objects in a crowded room. The device could be an adjunct to an electronic inventory system that tracks RFID-tagged objects as they move through reader-equipped doorways. Whereas commercial RFID-tag readers do not measure directions to tagged objects, the device is equipped with a phased-array antenna and a received signal-strength indicator (RSSI) circuit for measuring direction. At the beginning of operation, it is set to address only the RFID tag of interest. It then continuously transmits a signal to interrogate that tag while varying the radiation pattern of the antenna. It identifies the direction to the tag as the radiation pattern direction of peak strength of the signal returned by the tag. An approximate distance to the tag is calculated from the peak signal strength. The direction and distance can be displayed on a screen. A prototype containing a Yagi antenna was found to be capable of detecting a 915.5-MHz tag at a distance of approximately equal to 15 ft (approximately equal to 4.6 m).

  20. Gifted Male Readers: Current Understandings and Suggestions for Future Research

    ERIC Educational Resources Information Center

    Pagnani, Alexander R.

    2013-01-01

    Research literature concerning gifted male readers relies primarily on more extensive bodies of work regarding gifted males and male readers. Studied as a whole, the two halves portray a worrisome state of affairs for gifted male readers, who lag behind their female counterparts in the same patterns found across the ability spectrum. This literacy…

  1. 21 CFR 886.5810 - Ophthalmic prism reader.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Ophthalmic prism reader. 886.5810 Section 886.5810...) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5810 Ophthalmic prism reader. (a) Identification. An ophthalmic prism reader is a device intended for use by a patient who is in a supine...

  2. 21 CFR 886.5810 - Ophthalmic prism reader.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Ophthalmic prism reader. 886.5810 Section 886.5810...) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5810 Ophthalmic prism reader. (a) Identification. An ophthalmic prism reader is a device intended for use by a patient who is in a supine...

  3. 21 CFR 886.5810 - Ophthalmic prism reader.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Ophthalmic prism reader. 886.5810 Section 886.5810...) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5810 Ophthalmic prism reader. (a) Identification. An ophthalmic prism reader is a device intended for use by a patient who is in a supine...

  4. 21 CFR 886.5810 - Ophthalmic prism reader.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Ophthalmic prism reader. 886.5810 Section 886.5810...) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5810 Ophthalmic prism reader. (a) Identification. An ophthalmic prism reader is a device intended for use by a patient who is in a supine...

  5. 21 CFR 886.5810 - Ophthalmic prism reader.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Ophthalmic prism reader. 886.5810 Section 886.5810...) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5810 Ophthalmic prism reader. (a) Identification. An ophthalmic prism reader is a device intended for use by a patient who is in a supine...

  6. Reader Engagement in English and Persian Applied Linguistics Articles

    ERIC Educational Resources Information Center

    Ansarin, Ali Akbar; Tarlani-Aliabdi, Hassan

    2011-01-01

    There is an increasing interest in the way academic writers establish the presence of their readers over the past few years. Establishing the presence of readers or what Kroll (1984, p.181) calls imagining "a second voice" is accomplished when a writer refers "explicitly" to their readers using explicit linguistic resources…

  7. Tester automatically checks paper tape punch and reader after maintenance

    NASA Technical Reports Server (NTRS)

    Mazer, L.; Mc Murchy, D. D.

    1967-01-01

    Device automatically bench tests paper tape punches and readers in a simulated operating environment following routine maintenance. The reader and punch operate back-to-back and the paper tape output feeds the reader. The tape leader is prepunched with an arbitrary pattern that is continuously reproduced during the check.

  8. "Science in Society, Omnibus Pack, Readers M-P."

    ERIC Educational Resources Information Center

    Association for Science Education, Cambridge (England).

    Four additional readers have been written for use in the Science in Society general studies project. Three of the readers discuss the applications and importance of engineering in the world. They include: Engineering 1 (Reader M), which discusses such topics as the role of engineering in society, structural design and engineering, the engineering…

  9. A Comparison of Laterality Between Normal and Dyslexic Readers.

    ERIC Educational Resources Information Center

    Keefe, Barbara

    Examined with 38 right-handed boys who were either dyslexic or normal readers and matched for age and IQ (mean age both groups=10.6, mean IQ normal readers=106, mean IQ dyslexic readers=105) were the weak, strong, and equal lateralization theories of dyslexia. Cerebral lateralization was measured for linguistic material (digits) using the dichotic…

  10. Are You a Reader? 5th Graders Respond

    ERIC Educational Resources Information Center

    Barone, Diane; Barone, Rebecca

    2016-01-01

    The author tells the story of a 5th-grade teacher who challenges her class to take on self-identities as readers. Students defined seven characteristics of what it means to be a good reader and considered whether those characteristics applied to them: Good readers read for fun, talk about books, usually finish the book they're reading, can relate…

  11. The Language Skills and Concepts of Early and Nonearly Readers.

    ERIC Educational Resources Information Center

    Huba, M. E.; Ramisetty-Mikler, S.

    1995-01-01

    This study compared 56 early and nonearly readers (matched individually on sex, age, and preschool and kindergarten attended) in terms of their language concepts and skills, as well as their reading achievement, in kindergarten through second grade. The early readers were found superior to nonearly readers on preschool measures of general language…

  12. BEGINNING CHINESE READER, PART I. YALE LINGUISTIC SERIES.

    ERIC Educational Resources Information Center

    DE FRANCIS, JOHN; AND OTHERS

    THIS READER, ALONG WITH "BEGINNING CHINESE READER, PART II," IS CLOSELY INTEGRATED WITH THE SETON HALL UNIVERSITY TEXTS FOR SPOKEN AND WRITTEN CHINESE, "BEGINNING CHINESE" AND "CHARACTER TEXT FOR BEGINNING CHINESE." OF THE 400 CHARACTERS IN THESE TWO READERS, ALL BUT 33 HAVE BEEN PRESENTED TO THE STUDENT IN "BEGINNING CHINESE" AND "CHARACTER…

  13. HNO₃-assisted polyol synthesis of ultralarge single-crystalline Ag microplates and their far propagation length of surface plasmon polariton.

    PubMed

    Chang, Cheng-Wei; Lin, Fan-Cheng; Chiu, Chun-Ya; Su, Chung-Yi; Huang, Jer-Shing; Perng, Tsong-Pyng; Yen, Ta-Jen

    2014-07-23

    We developed a HNO3-assisted polyol reduction method to synthesize ultralarge single-crystalline Ag microplates routinely. The edge length of the synthesized Ag microplates reaches 50 μm, and their top facets are (111). The mechanism for dramatically enlarging single-crystalline Ag structure stems from a series of competitive anisotropic growths, primarily governed by carefully tuning the adsorption of Ag(0) by ethylene glycol and the desorption of Ag(0) by a cyanide ion on Ag(100). Finally, we measured the propagation length of surface plasmon polaritons along the air/Ag interface under 534 nm laser excitation. Our single-crystalline Ag microplate exhibited a propagation length (11.22 μm) considerably greater than that of the conventional E-gun deposited Ag thin film (5.27 μm).

  14. Quantifying protein by bicinchoninic Acid.

    PubMed

    Simpson, Richard J

    2008-08-01

    INTRODUCTIONThis protocol describes a method of quantifying protein that is a variation of the Lowry assay. It uses bicinchoninic acid (BCA) to enhance the detection of Cu(+) generated under alkaline conditions at sites of complexes between Cu(2+) and protein. The resulting chromophore absorbs at 562 nm. This technique is divided into three parts: Standard Procedure, Microprocedure, and 96-Well Microtiter Plate Procedure. For each procedure, test samples are assayed in parallel with protein standards that are used to generate a calibration curve, and the exact concentration of protein in the test samples is interpolated. The standard BCA assay uses large volumes of both reagents and samples and cannot easily be automated. If these issues are important, the Microprocedure is recommended. This in turn can be adapted for use with a microplate reader in the 96-Well Microtiter Plate Procedure. If the microplate reader is interfaced with a computer, more than 1000 samples can be read per hour.

  15. An optimized microplate assay system for quantitative evaluation of plant cell wall-degrading enzyme activity of fungal culture extracts.

    PubMed

    King, Brian C; Donnelly, Marie K; Bergstrom, Gary C; Walker, Larry P; Gibson, Donna M

    2009-03-01

    Developing enzyme cocktails for cellulosic biomass hydrolysis complementary to current cellulase systems is a critical step needed for economically viable biofuels production. Recent genomic analysis indicates that some plant pathogenic fungi are likely a largely untapped resource in which to prospect for novel hydrolytic enzymes for biomass conversion. In order to develop high throughput screening assays for enzyme bioprospecting, a standardized microplate assay was developed for rapid analysis of polysaccharide hydrolysis by fungal extracts, incorporating biomass substrates. Fungi were grown for 10 days on cellulose- or switchgrass-containing media to produce enzyme extracts for analysis. Reducing sugar released from filter paper, Avicel, corn stalk, switchgrass, carboxymethylcellulose, and arabinoxylan was quantified using a miniaturized colorimetric assay based on 3,5-dinitrosalicylic acid. Significant interactions were identified among fungal species, growth media composition, assay substrate, and temperature. Within a small sampling of plant pathogenic fungi, some extracts had crude activities comparable to or greater than T. reesei, particularly when assayed at lower temperatures and on biomass substrates. This microplate assay system should prove useful for high-throughput bioprospecting for new sources of novel enzymes for biofuel production.

  16. Microplate fluorescence protease assays test the inhibition of select North American snake venoms' activities with an anti-proteinase library.

    PubMed

    Price, Joseph A

    2015-09-01

    Snake envenomation is a relatively neglected significant world health problem, designated an orphan disease by the WHO. While often effective, antivenins are insufficient. Could another approach greatly aid inhibition of the venom toxins? New fluorescent substrates for measuring protease activity in microplate assays suitable for high throughput screening were tested and found reproducible with snake venom. Representative North American venoms showed relatively strong proteinase and collagenase, but weaker elastase activities. Caseinolytic activity is inhibited by the nonspecific proteinase inhibitor 1,10-phenanthroline and by EDTA, as is collagenase activity, consistent with the action of metalloproteinases. Both general protease and collagenase assays CV average 3%, and Km measured were above normal working conditions. Using a library of anti -proteinase compounds with multiple venoms revealed high inhibitor activity by three agents with known multiple metalloproteinase inhibitor activity (Actinonin, GM6001, and NNGH), which incidentally supports the concept that much of the degradative activity of certain venoms is due to metalloproteinases with collagenase activity. These results together support the use of microplate proteinase assays, particularly this collagenase assay, in future drug repurposing studies leading to the development of new treatments for those envenomations that have a major proteolytic component in their pathophysiology.

  17. Space-time model for migration of weak earthquakes along the northern boundary of the Amurian microplate

    NASA Astrophysics Data System (ADS)

    Trofimenko, S. V.; Bykov, V. G.; Merkulova, T. V.

    2016-07-01

    In this paper, we aimed to investigate the statistical distributions of shallow earthquakes with 2 ≤ M ≤ 4, located in 13 rectangular areas (clusters) bounded by 120°E and 144°E along the northern boundary of the Amurian microplate. As a result of our study, the displacement of seismicity maxima has been determined and three recurrent spatial cycles have been observed. The clusters with similar distribution of earthquakes are suggested to alternate being equally spaced at 7.26° (360-420 km). A comparison of investigation results on the structure of seismicity in various segments of the Amurian microplate reveals the identity between the alternation pattern observed for meridional zones of large earthquakes and a distinguished spatial period. The displacement vector for seismicity in the annual cycles is determined, and the correspondence between its E-W direction and the displacement of the fronts of large earthquakes is established. The elaborated model of seismic and deformation processes is considered, in which subsequent activation of clusters of weak earthquakes (2 ≤ M ≤ 4), tending to extend from the Japanese-Sakhalin island arc to the eastern closure of the Baikal rift zone, is initiated by the displacement of the strain wave front.

  18. High-sensitive and high-efficient biochemical analysis method using a bionic electronic eye in combination with a smartphone-based colorimetric reader system.

    PubMed

    Kaiqi Su; Quchao Zou; Ning Hu; Ping Wang

    2015-01-01

    Bionic electronic eye (Bionic e-Eye), a developed smartphone-based colorimetric reader system, consists of smartphone or pad (iPhone 4s or iPad 3) as detection instrument and portable accessory as illumination provider, integrating with a wide-angle lens, a piece of lowest-power electro luminescent and a custom-made dark hood. A 96-well microtiter plate (MTP) was positioned on the electro luminescent and Bionic e-Eye captures the detection images by the back camera of smartphone. Being similar to human visual system, the hue, saturation and value (HSV, also called hex cone model) color model was employed in image processing algorithm of Bionic e-Eye. Optimized system dimension was determined by the system steadiness experiment of different photograph distances. Moreover, the commercially available BCA protein assay and CCK8 cell number assay were carried out to evaluate this Bionic e-Eye. Analytical performance of Bionic e-Eye had the better precision, higher sensitivity than microtiter plate reader (MTPR) and previous smartphone-based colorimetric reader for both two assays. Also, Bionic e-Eye using optical image detection had simultaneous and synchronous working mode, while MPTR using machine moving detection had asynchronous working mode in high throughput detection. Therefore, Bionic e-Eye will be an ideal point-of-care (POC) colorimetric detection device in the field of clinical application, industrial quality control, environment monitoring, and food assessment.

  19. High-throughput, semi-automated determination of a cyclooxygenase II inhibitor in human plasma and urine using solid-phase extraction in the 96-well format and high-performance liquid chromatography with post-column photochemical derivatization-fluorescence detection.

    PubMed

    Matthews, C Z; Woolf, E J; Lin, L; Fang, W; Hsieh, J; Ha, S; Simpson, R; Matuszewski, B K

    2001-02-25

    Compound I, 5-chloro-3-(4-methanesulfonylphenyl)-6'-methyl-[2,3']bipyridinyl, has been found to be a specific inhibitor of the enzyme cyclooxygenase II (COX II). The anti-inflammatory properties of this compound are currently being investigated. HPLC assays for the determination of this analyte in human plasma and human urine have been developed. Isolation of I and the internal standard (II) was achieved by solid-phase extraction (SPE) in the 96-well format. A C8 SPE plate was used for the extraction of the drug from human plasma (recovery >90%) while a mixed-mode (C8/Cation) SPE plate was used to isolate the analytes from human urine (recovery approximately 71%). The analyte and internal standard were chromatographed on a Keystone Scientific Prism-RP guard column (20 x 4.6 mm) connected to a Prism-RP analytical column (150 x 4.6 mm), using a mobile phase consisting of 45% acetonitrile in 10 mM acetate buffer (pH = 4); the analytes eluted at retention times of 5.2 and 6.9 min for I and II, respectively. Compounds I and II were found to form highly fluorescent products after exposure to UV light (254 nm). Thus, the analytes were detected by fluorescence (lambda(ex) = 260 nm, lambda(em) =375 nm) following post-column photochemical derivatization. Eight point calibration curves over the concentration range of 5-500 ng/ml for human plasma and human urine yielded a linear response (R2>0.99) when a 1/y weighted linear regression model was employed. Based on the replicate analyses (n = 5) of spiked standards, the within-day precision for both assays was better than 7% C.V. at all points on the calibration curve; within-day accuracy was within 5% of nominal at all standard concentrations. The between-run precision and accuracy of the assays, as calculated from the results of the analysis of quality control samples, was better than 8% C.V. and within 8% of nominal. I was found to be stable in human plasma and urine for at least 8 and 2 months, respectively. In addition, the

  20. AGU Publications: Improvements for Authors and Readers

    NASA Astrophysics Data System (ADS)

    Hanson, Brooks

    2013-12-01

    AGU has introduced several new features aimed at simplifying and improving the submission of papers to AGU journals. Enhanced PDF and HTML formats and new journal home pages developed with our publishing partner, Wiley, will also provide improvements for readers. In previous issues of Eos, we provided broader overviews of AGU publications, including the transition to Wiley and open access (Eos, 94(30), 264-266, doi:10.1002/2013EO300009; Eos, 94(39), 345, doi:10.1002/2013EO390006).

  1. BOOK REVIEW: Astrophysics (Advanced Physics Readers)

    NASA Astrophysics Data System (ADS)

    Kibble, Bob

    2000-07-01

    Here is a handy and attractive reader to support students on post-16 courses. It covers the astrophysics, astronomy and cosmology that are demanded at A-level and offers anyone interested in these fields an interesting and engaging reference book. The author and the production team deserve credit for producing such an attractive book. The content, in ten chapters, covers what one would expect at this level but it is how it is presented that struck me as the book's most powerful asset. Each chapter ends with a summary of key ideas. Line drawings are clear and convey enough information to make them more than illustrations - they are as valuable as the text in conveying information. Full colour is used throughout to enhance illustrations and tables and to lift key sections of the text. A number of colour photographs complement the material and serve to maintain interest and remind readers that astrophysics is about real observable phenomena. Included towards the end is a set of tables offering information on physical and astronomical data, mathematical techniques and constellation names and abbreviations. This last table puzzled me as to its value. There is a helpful bibliography which includes society contacts and a website related to the text. Perhaps my one regret is that there is no section where students are encouraged to actually do some real astronomy. Astrophysics is in danger of becoming an armchair and calculator interest. There are practical projects that students could undertake either for school assessment or for personal interest. Simple astrophotography to capture star trails, observe star colours and estimate apparent magnitudes is an example, as is a simple double-star search. There are dozens more. However, the author's style is friendly and collaborative. He befriends the reader as they journey together through the ideas. There are progress questions at the end of each chapter. Their style tends to be rather closed and they emphasize factual recall

  2. Analysis of Able and Disabled Sixth-Grade Readers' Knowledge of Story Structure: A Comparison.

    ERIC Educational Resources Information Center

    Krein, Evelyn Leech; Zaharias, Jane Ann

    1986-01-01

    Confirms that able readers have a more well-rounded sense of story structure than disabled readers. Specifically, that able readers demonstrated an ability to tell more elaborate stories than disabled readers. (FL)

  3. Moho-Depth Dike Swarms and Rifting of the Sierra Nevada Microplate, Northeast California

    NASA Astrophysics Data System (ADS)

    Smith, K. D.; Kent, G.; von Seggern, D. H.; Eissis, A.; Driscoll, N. W.

    2012-12-01

    Moho-depth earthquake swarms below N. Lake Tahoe (LT) in late 2003 and, beneath southern Sierra Valley (SV), California in late 2011/early 2012, ~50 km to the north, define nearly identical dike injection events along the same Moho-depth structure. Each swarm lasts ~6 months, outlines an ~7x7 km fault area, initiates at its deepest extent, shows dominantly reverse slip mechanisms, and aligns along an ~N45W striking, ~50 degree east dipping Moho-depth structure. About 1600 events were located in the LT swarm (maximum M 2.2; b-value 2.0) and nearly 2200 have been located under SV (maximum M 1.9; b-value 1.7); an NSF-Earthscope supported deployment in SV includes five broadband instruments that are currently operating within one focal depth - three directly above the sequence. A Long-Period earthquake (LP), located at a depth of 30 km along the dipping structure (9/29/2011) was located midway between the two sequences. This is the first LP identified outside of the Long Valley volcanic center along the northeastern Sierra. These diking events define a northward propagating rift boundary along the northeastern boundary of the Sierran Microplate. Extension driven decompression melting of upper mantle magmas and weakening of a stronger (~5 km thick) upper-Moho lid drives dike injection and faulting of the rift structure. Following failure of the strong upper mantle layer, footwall unloading and flexural response help bring the Sierran block elevations into equilibrium with the upper-mantle and crustal profile. Forces driving plate boundary processes, slab roll back, westward escape of the southern Cascadia upper plate, and northward evolution of the Mendocino Triple Junction (MTJ) result in northwestward motion of the Sierra block (e.g., Atwater and Stock, 1988) and extension east of the Sierra, driving northwestward rift propagation. These overall processes are reflected in the historical earthquake record in northeast California and western Nevada. Shear dominated San

  4. Metacognition and Reading: Comparing Three Forms of Metacognition in Normally Developing Readers and Readers with Dyslexia

    PubMed Central

    Brunswick, Nicola

    2015-01-01

    Metacognition refers to ‘cognition about cognition’ and includes metacognitive knowledge, strategies and experiences (Efklides, 2008; Flavell, 1979). Research on reading has shown that better readers demonstrate more metacognitive knowledge than poor readers (Baker & Beall, 2009), and that reading ability improves through strategy instruction (Gersten, Fuchs, Williams, & Baker, 2001). The current study is the first to specifically compare the three forms of metacognition in dyslexic (N = 22) versus normally developing readers (N = 22). Participants read two factual texts, with learning outcome measured by a memory task. Metacognitive knowledge and skills were assessed by self-report. Metacognitive experiences were measured by predictions of performance and judgments of learning. Individuals with dyslexia showed insight into their reading problems, but less general knowledge of how to approach text reading. They more often reported lack of available reading strategies, but groups did not differ in the use of deep and surface strategies. Learning outcome and mean ratings of predictions of performance and judgments of learning were lower in dyslexic readers, but not the accuracy with which metacognitive experiences predicted learning. Overall, the results indicate that dyslexic reading and spelling problems are not generally associated with lower levels of metacognitive knowledge, metacognitive strategies or sensitivity to metacognitive experiences in reading situations. @ 2015 The Authors. Dyslexia published by John Wiley & Sons Ltd. PMID:26234622

  5. Error Detection Mechanism for Words and Sentences: A Comparison between Readers with Dyslexia and Skilled Readers

    ERIC Educational Resources Information Center

    Horowitz-Kraus, Tzipi; Breznitz, Zvia

    2011-01-01

    The activity level of the error monitoring system for processing isolated versus contextual words in Hebrew was studied in adults with dyslexia and skilled readers while committing reading errors. Behavioural measures and event-related potentials were measured during a lexical decision task using words in a list and sentences. Error-related…

  6. Metacognition and Reading: Comparing Three Forms of Metacognition in Normally Developing Readers and Readers with Dyslexia.

    PubMed

    Furnes, Bjarte; Norman, Elisabeth

    2015-08-01

    Metacognition refers to 'cognition about cognition' and includes metacognitive knowledge, strategies and experiences (Efklides, 2008; Flavell, 1979). Research on reading has shown that better readers demonstrate more metacognitive knowledge than poor readers (Baker & Beall, 2009), and that reading ability improves through strategy instruction (Gersten, Fuchs, Williams, & Baker, 2001). The current study is the first to specifically compare the three forms of metacognition in dyslexic (N = 22) versus normally developing readers (N = 22). Participants read two factual texts, with learning outcome measured by a memory task. Metacognitive knowledge and skills were assessed by self-report. Metacognitive experiences were measured by predictions of performance and judgments of learning. Individuals with dyslexia showed insight into their reading problems, but less general knowledge of how to approach text reading. They more often reported lack of available reading strategies, but groups did not differ in the use of deep and surface strategies. Learning outcome and mean ratings of predictions of performance and judgments of learning were lower in dyslexic readers, but not the accuracy with which metacognitive experiences predicted learning. Overall, the results indicate that dyslexic reading and spelling problems are not generally associated with lower levels of metacognitive knowledge, metacognitive strategies or sensitivity to metacognitive experiences in reading situations.

  7. Proficient Readers' Reading Behavior in Taiwan: The Study of Young Chinese Readers

    ERIC Educational Resources Information Center

    Chang, Li-Chun

    2015-01-01

    The purpose of this study was to explore the reading behavior of young proficient Chinese readers at preschool age. Especially, the roles of phonetic skill and Chinese Character recognition in reading comprehension were explored. 10 kindergartens were recruited to participate in the study. Subjects were 72-98 kindergarten children. Instruments…

  8. Reader Development Bibliography. Books Recommended for Adult New Readers. Fourth Edition.

    ERIC Educational Resources Information Center

    Collins, Vickie L.

    This edition is an annotated list of 390 literacy materials for adult new readers. All the materials are written on an eighth-grade level or below. These paperback instructional titles, many in workbook format, are for use with adult basic education (ABE) and English-as-a-Second-Language (ESL) students. The introduction includes guidelines for…

  9. The Reader-Text-Writer Interaction: L2 Japanese Learners' Response toward Graded Readers

    ERIC Educational Resources Information Center

    Tabata-Sandom, Mitsue

    2013-01-01

    This paper reports on two projects which investigated graded readers (GRs) as meaningful input for learners of Japanese as a foreign language (JFL). Project One examined the intentions of six writers of Japanese GRs. A focus group interview demonstrated that the writers had a genuine communicative intent in the writing process. Project Two…

  10. Late Quaternary Activity and Seismogenic Potential of the Gonave Microplate: Plantain Garden Strike-Slip Fault Zone of Eastern Jamaica

    NASA Astrophysics Data System (ADS)

    Mann, P.; Prentice, C.; King, W.; Demets, C.; Wiggins-Grandison, M.; Benford, B.

    2008-12-01

    At the longitude of Jamaica, Caribbean (Carib)-North America (Noam) plate motion of 19 ± 2 mm/a is carried by two parallel, left-lateral strike-slip faults, the Oriente fault zone, immediately south of Cuba, and the Enriquillo-Plantain Garden fault zone (EPGFZ), which lies 100-150 km further south. It has been postulated that the lithosphere between these faults constitutes an independent Gonave microplate that has formed in response to the ongoing collision between the leading edge of Carib in Hispaniola and the Bahama carbonate platform. GPS measurements in Jamaica and Hispanola is supportive of the microplate hypothesis and indicates that roughly half of Carib-Noam plate motion (8-14 mm/a) is carried by the EPGFZ of southern Hispaniola and eastern Jamaica. This study applies geomorphic and paleoseismic methods as a direct test of the activity and amount of microplate motion carried on the Plantain Garden fault segment of eastern Hispaniola and how this motion is distributed across a large restraining bend that has formed the island of Jamaica since the late Miocene. The EPFZ curves gently to the northeast and forming a steep mountain front to the Blue Mountains restraining bend with elevations up to 2200 m. Geomorphic fault-related features along the mountain front fault zone include left-laterally deflected rivers and streams, but no small scale features indicative of Holocene activity. River and stream deflections range from 0.1 to 0.5 km. We identified and trenched the most active trace of the mountain front fault at the Morant River where the fault is characterized by a 1.5-m-wide sub-vertical fault zone juxtaposing sheared alluvium and fault Cretaceous basement rocks This section is overlain by a 6-m-thick fluvial terrace. Trenching in the unfaulted terrace immediately overlying the fault trace revealed radiocarbon and OSL ages ranging from 20 to 21 ka that are consistent with a prominent unfaulted alluvial fan along the projection of this fault 1.5 km to

  11. Crustal Structure of the Yakutat Microplate: New Parameters for Understanding the Evolution of the Chugach-St.Elias Orogeny

    NASA Astrophysics Data System (ADS)

    Worthington, L. L.; Christeson, G. L.; van Avendonk, H. J.; Gulick, S. P.

    2009-12-01

    We present results of a 2008 marine seismic reflection/refraction survey acquired as part of the St. Elias Erosion and Tectonics Project (STEEP), a multi-disciplinary NSF-Continental Dynamics project aimed at tectonic-climate interaction, structural evolution and geodynamics in the Chugach-St. Elias orogen. The Chugach-St.Elias orogen is the result of flat-slab subduction and collision of the Yakutat (YAK) microplate with North Amercian (NA) on the southern Alaska margin during the last ~10Ma. A fundamental goal of STEEP is to address controversy related to the deep crustal structure of the YAK block itself, describe its offshore structural relationships and constrain its buoyancy in order to understand the orogenic driver. Marine seismic reflection profiles acquired across the offshore YAK microplate provide the first regional images of the top of the subducting YAK basement. The basement reflector is observed near the seafloor at the Dangerous River Zone (DRZ) and is overlain by up to 12 km of sediments near Kayak Island, resulting in a basement dip of ~3° in the direction of subduction. The basement reflector also shallows near the shelf-edge adjacent to the Transition Fault, the YAK-Pacific boundary. These observations are indicative of an overall regional basement tilt towards the NA continent. Two coincident wide-angle refraction profiles constrain YAK crustal thickness between 30-35km, >20km thicker than normal oceanic crust, and lower crustal velocities potentially >7km/s. Crustal velocity and thickness are comparable to the Kerguelen oceanic plateau and the Siletz terrane. These results are the first direct observations in support of the oceanic plateau theory for the origin of the YAK microplate. Crustal velocity and structure are continuous across the DRZ on the YAK shelf, which is historically described as a vertical boundary between continental crust on the east and oceanic basement on the west. Instead, we observe a gradual shallowing of elevated

  12. Low-cost and highly efficient DNA biosensor for heavy metal ion using specific DNAzyme-modified microplate and portable glucometer-based detection mode.

    PubMed

    Zhang, Jin; Tang, Ying; Teng, Liumei; Lu, Minghua; Tang, Dianping

    2015-06-15

    A simple and low-cost DNA sensing platform based on Pb(2+)-specific DNAzyme-modified microplate was successfully developed for highly sensitive monitoring of lead ion (Pb(2+), one kind of toxic heavy metal ion) in the environmental samples coupling with a portable personal glucometer (PGM)-based detection mode. The detection cell was first prepared simply by means of immobilizing the DNAzyme on the streptavidin-modified microplate. Gold nanoparticle labeled with single-stranded DNA and invertase (Enz-AuNP-DNA) was utilized as the signal-transduction tag to produce PGM substrate (glucose). Upon addition of lead ion into the microplate, the substrate strand of the immobilized DNAzyme was catalytically cleaved by target Pb(2+), and the newly generated single-strand DNA in the microplate could hybridize again with the single-stranded DNA on the Enz-AuNP-DNA. Accompanying with the Enz-AuNP-DNA, the carried invertase could convert sucrose into glucose. The as-produced glucose could be monitored by using a widely accessible PGM for in situ amplified digital readout. Based on Enz-AuNP-DNA amplification strategy, as low as 1.0 pM Pb(2+) could be detected under the optimal conditions. Moreover, the methodology also showed good reproducibility and high selectivity toward target Pb(2+) against other metal ions because of highly specific Pb(2+)-dependent DNAzyme, and was applicable for monitoring Pb(2+) in the naturally contaminated sewage and spiked drinking water samples.

  13. A rapid and specific microplate assay for the determination of intra- and extracellular ascorbate in cultured cells.

    PubMed

    Lane, Darius J R; Lawen, Alfons

    2014-04-11

    Vitamin C (ascorbate) plays numerous important roles in cellular metabolism, many of which have only come to light in recent years. For instance, within the brain, ascorbate acts in a neuroprotective and neuromodulatory manner that involves ascorbate cycling between neurons and vicinal astrocytes--a relationship that appears to be crucial for brain ascorbate homeostasis. Additionally, emerging evidence strongly suggests that ascorbate has a greatly expanded role in regulating cellular and systemic iron metabolism than is classically recognized. The increasing recognition of the integral role of ascorbate in normal and deregulated cellular and organismal physiology demands a range of medium-throughput and high-sensitivity analytic techniques that can be executed without the need for highly expensive specialist equipment. Here we provide explicit instructions for a medium-throughput, specific and relatively inexpensive microplate assay for the determination of both intra- and extracellular ascorbate in cell culture.

  14. A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells

    PubMed Central

    Lane, Darius J. R.; Lawen, Alfons

    2014-01-01

    Vitamin C (ascorbate) plays numerous important roles in cellular metabolism, many of which have only come to light in recent years. For instance, within the brain, ascorbate acts in a neuroprotective and neuromodulatory manner that involves ascorbate cycling between neurons and vicinal astrocytes - a relationship that appears to be crucial for brain ascorbate homeostasis. Additionally, emerging evidence strongly suggests that ascorbate has a greatly expanded role in regulating cellular and systemic iron metabolism than is classically recognized. The increasing recognition of the integral role of ascorbate in normal and deregulated cellular and organismal physiology demands a range of medium-throughput and high-sensitivity analytic techniques that can be executed without the need for highly expensive specialist equipment. Here we provide explicit instructions for a medium-throughput, specific and relatively inexpensive microplate assay for the determination of both intra- and extracellular ascorbate in cell culture. PMID:24747535

  15. Fluorescence anisotropy microplate assay to investigate the interaction of full-length steroid receptor coactivator-1a with steroid receptors

    PubMed Central

    Zhang, Chen; Nordeen, Steven K.; Shapiro, David J.

    2013-01-01

    Estrogens, acting via estrogen receptor (ER) play key roles in growth, differentiation and gene regulation in the reproductive, central nervous and skeletal systems. ER-mediated gene transcription contributes to the development and spread of breast, uterine, and liver cancer. Steroid receptor coactivator-1a (SRC1a) belongs to the P160 family of coactivators, which is the best known of the many coactivators implicated in ER-mediated transactivation. Binding of full-length P160 coactivators to steroid receptors has been difficult to investigate in vitro. This chapter details how to investigate the interaction of SRC1a with ER using the fluorescence anisotropy/polarization microplate assay (FAMA). PMID:23436375

  16. Large-scale preparation of shape controlled SnO and improved capacitance for supercapacitors: from nanoclusters to square microplates

    NASA Astrophysics Data System (ADS)

    Wang, Lu; Ji, Hongmei; Zhu, Feng; Chen, Zhi; Yang, Yang; Jiang, Xuefan; Pinto, João; Yang, Gang

    2013-07-01

    Here, we first provide a facile ultrasonic-assisted synthesis of SnO using SnCl2 and the organic solvent of ethanolamine (ETA). The moderate alkalinity of ETA and ultrasound play very important roles in the synthesis of SnO. After the hydrolysis of the intermediate of ETA-Sn(ii), the as-synthesized SnO nanoclusters undergo assembly, amalgamation, and preferential growth to microplates in hydrothermal treatment. The as-synthesized SnO was characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), ultraviolet-visible absorption spectroscopy (UV-vis) and X-ray diffraction (XRD). To explore its potential applications in energy storage, SnO was fabricated into a supercapacitor electrode and characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and galvanostatic charge-discharge measurements. The as-synthesized SnO exhibits remarkable pseudocapacitive activity including high specific capacitance (208.9 F g-1 at 0.1 A g-1), good rate capability (65.8 F g-1 at 40 A g-1), and excellent cycling stability (retention 119.3% after 10 000 cycles) for application in supercapacitors. The capacitive behavior of SnO with various crystal morphologies was observed by fitted EIS using an equivalent circuit. The novel synthetic route for SnO is a convenient and potential way to large-scale production of microplates which is expected to be applicable in the synthesis of other metal oxide nanoparticles.Here, we first provide a facile ultrasonic-assisted synthesis of SnO using SnCl2 and the organic solvent of ethanolamine (ETA). The moderate alkalinity of ETA and ultrasound play very important roles in the synthesis of SnO. After the hydrolysis of the intermediate of ETA-Sn(ii), the as-synthesized SnO nanoclusters undergo assembly, amalgamation, and preferential growth to microplates in hydrothermal treatment. The as-synthesized SnO was characterized by scanning

  17. Neutral red cytotoxicity assays for assessing in vivo carbon nanotube ecotoxicity in mussels--Comparing microscope and microplate methods.

    PubMed

    Miller, M A; Bankier, C; Al-Shaeri, M A M; Hartl, M G J

    2015-12-30

    The purpose of the present study was to compare two neutral red retention methods, the more established but very labour-intensive microscope method (NRR) against the more recently developed microplate method (NRU). The intention was to explore whether the sample volume throughput could be increased and potential operator bias avoided. Mussels Mytilus sp. were exposed in vivo to 50, 250 and 500 μg L(-1) single (SWCNTs) or multi-walled carbon nanotubes (MWCNTs). Using the NRR method, SWCNTs and MWCNTs caused concentration dependent decreases in neutral red retention time. However, a concentration dependent decrease in optical density was not observed using the NRU method. We conclude that the NRU method is not sensitive enough to assess carbon nanotube ecotoxicity in vivo in environmentally relevant media, and recommend using the NRR method.

  18. U-Pb isotopic evidence for accretion of a continental microplate in the Zalm region of the Saudi Arabian Shield

    USGS Publications Warehouse

    Stacey, J.S.; Agar, R.A.

    1986-01-01

    U-Pb zircon age determinations show that this "Andean" continental margin developed before about 720 Ma, and emplacement of calc-alkaline plutonic rocks continued until about 690 Ma. During the period 690-640 Ma, the continental Afif microplate collided with the Asir terrane as part of the Nabitah orogeny. At approximately 640 Ma ago, the Najd strike-slip orogen commenced with a dextral phase that controlled the emplacement of granitic plutons as well as the development of a series of large pull-apart grabens. Some of these grabens were floored by new oceanic crust and were filled with volcano-sedimentary rocks of the Bani Ghayy group. Subsequently, the Najd fault system changed to sinistral strike slip motion at about 620 Ma ago.

  19. Neural syntax: cell assemblies, synapsembles and readers

    PubMed Central

    Buzsáki, György

    2010-01-01

    Summary A widely discussed hypothesis in neuroscience is that transiently active ensembles of neurons, known as ‘cell assemblies’, underlie numerous operations of the brain, from encoding memories to reasoning. However, the mechanisms responsible for the formation and disbanding of cell assemblies and temporal evolution of cell assembly sequences are not well understood. I introduce and review three interconnected topics, which could facilitate progress in defining cell assemblies, identifying their neuronal organization and revealing causal relationships between assembly organization and behavior. First, I hypothesize that cell assemblies are best understood in light of their output product, as detected by ‘reader-actuator’ mechanisms. Second, I suggest that the hierarchical organization of cell assemblies may be regarded as a neural syntax. Third, constituents of the neural syntax are linked together by dynamically changing constellations of synaptic weights (‘synapsembles’). Existing support for this tripartite framework is reviewed and strategies for experimental testing of its predictions are discussed. PMID:21040841

  20. Early implications of the COCONet GPS velocity field for studies of plate and microplate motions in the Caribbean

    NASA Astrophysics Data System (ADS)

    DeMets, C.

    2013-05-01

    Now entering their 3rd decade, GPS measurements in the Caribbean region have been used to study a wide range of tectonic topics such as the movement of the Caribbean plate relative to North and South America, earthquake cycle effects along the seismically hazardous Caribbean plate boundary faults, and microplate interactions along the complexly deforming Caribbean plate boundaries. The construction of COCONet stands out as the most significant-ever one-time advance in GPS-MET infrastructure in the Caribbean region due to its standardized GPS-MET equipment, its open access to real-time data, and its expansion of coverage relative to pre-existing GPS stations. In this talk, I will show an up-to-date Caribbean region velocity field derived using the most recent version of GIPSY software (release 6.1), the latest satellite orbit products, single-station ambiguity resolution, and a consistent realization of the Caribbean plate reference frame. Consisting of several hundred site velocities, the new velocity field clearly defines how the crust responds to east-to-west changes in the geometry of the Caribbean-North America plate boundary, including the profound effect of the oblique collision zone between the Bahama platform and northern edge of the Caribbean plate. I will discuss implications of the still-maturing COCONet GPS velocities for ongoing studies of Caribbean plate motion and plate rigidity and will also discuss applications of COCONet velocities for testing recently published kinematic estimates for the movements of the Gonave, Puerto Rico, Hispaniola, and southern Jamaica microplates.

  1. Development and testing of a multiwell plates absorbance reader for clinical analysis using inexpensive webcam

    NASA Astrophysics Data System (ADS)

    Castillo, Jimmy; Gutierrez, Hector; Vitta, Yosmery; Martinez, Mauro; Fernandez, Alberto

    2007-09-01

    Biochemical analysis and clinical tests like glucose, hemoglobin, cholesterol, iron, etc. are crucial for early illness diagnosis like diabetes, anemia and coronary deceases. These tests usually are done in state of the art instruments in well equipped laboratories in health centers. In some cases, these instruments are not portable, so they are not recommended for clinical field studies in remote areas. The present work shows a portable low-cost prototype of multi-well plates reader designed for clinical analysis. A Light Emission Diodes (LEDs) array is used as excitation source and an inexpensive webcam as detector. The light source illuminates the 96 well plates and the webcam take the image with 640x480 pixels. The data is acquired and processed by using a portable computer. 96 samples can be read including blanks and calibration standards simultaneously. Light absorption data are processed using a MatLab software designed in our laboratory to obtain calibration curves, standards lectures and samples concentration. The system was evaluated using different analytes series solutions: Neutral Red, Cooper (II) Ammonia Complex and Methyl Orange. The results shows that it is possible to measure few micro liters of solutions with adequate exactitude and precision of less than 3%. As possible analytical clinical application, iron determination was performed using Fe(III) Thiocyanate complex. This method is usually applied in serum samples analysis. The sensibility achieved with the proposed instrumentation configurations allows the analysis of iron in serum samples in the references values normal range (0.75 - 1.5 mg/L) in human.

  2. What do mind readers know and what do we know about mind readers?

    PubMed

    Woolley, Jacqueline D

    2014-11-01

    In this commentary, I raise various questions about Kim and Harris's fascinating findings. I ask what kind of knowledge children expect telepathic individuals to have, who children might consider to be good mind readers, why children value telepathy, and how puzzled children are by telepathy. I suggest potential ways to address some of these questions and end by reiterating the importance of probing individual differences in scepticism and credulity.

  3. Motivating Struggling Adolescent Readers: An Action Research Study

    ERIC Educational Resources Information Center

    Richardson, Eileen M.

    2016-01-01

    In a high-school reading class, the author used Reader's Theater as an instructional and motivational strategy for underachieving students. This action research focused on the extent to which implementing Reader's Theater motivated students to read and improve their reading skills. Consistent increases in scores for all students occurred over the…

  4. 21 CFR 886.5800 - Ophthalmic bar reader.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Ophthalmic bar reader. 886.5800 Section 886.5800 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5800 Ophthalmic bar reader....

  5. 21 CFR 886.5800 - Ophthalmic bar reader.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Ophthalmic bar reader. 886.5800 Section 886.5800 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5800 Ophthalmic bar reader....

  6. 21 CFR 886.5800 - Ophthalmic bar reader.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Ophthalmic bar reader. 886.5800 Section 886.5800 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5800 Ophthalmic bar reader....

  7. 21 CFR 886.5800 - Ophthalmic bar reader.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Ophthalmic bar reader. 886.5800 Section 886.5800 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5800 Ophthalmic bar reader....

  8. 21 CFR 886.5800 - Ophthalmic bar reader.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Ophthalmic bar reader. 886.5800 Section 886.5800 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5800 Ophthalmic bar reader....

  9. Prevalence and Nature of Late-Emerging Poor Readers

    ERIC Educational Resources Information Center

    Catts, Hugh W.; Compton, Donald; Tomblin, J. Bruce; Bridges, Mindy Sittner

    2012-01-01

    Some children demonstrate adequate or better reading achievement in early school grades but fall significantly behind their peers in later grades. These children are often referred to as late-emerging poor readers. In this study, we investigated the prevalence and heterogeneity of these poor readers. We also examined the early language and…

  10. Learning to Choose: The Hidden Art of the Enthusiastic Reader

    ERIC Educational Resources Information Center

    Mackey, Margaret

    2014-01-01

    Often the difference between a successful and unsuccessful reader is that the former is simply better at finding enjoyable books to read. The capacity to select appealing reading is not developed in classrooms where the decision about what book to read is made by someone other than the reader: the teacher, the curriculum planner, the person who…

  11. Did Anything Happen?: Chance Composition in Readers Theatre.

    ERIC Educational Resources Information Center

    Bindert, Kathleen

    During the 1950s, when it first began, Readers Theatre was textual in its emphasis; it involved the group reading of literature and the staging of poems and poets' stories, essays, and scenes. In its second stage, during the sixties, Readers Theatre became a directors' art; it was the director who adapted and arranged the literature and whose…

  12. Accelerated Reader/Reading Renaissance. What Works Clearinghouse Intervention Report

    ERIC Educational Resources Information Center

    What Works Clearinghouse, 2007

    2007-01-01

    The Accelerated Reader/Reading Renaissance program (now called Accelerated Reader Best Classroom Practices) is a guided reading intervention in which teachers direct student reading of text. It involves two components. Reading Renaissance, the first component, is a set of recommended principles on guided reading (or teachers' direction of…

  13. Skilled or Skillful: What's the Difference for Readers?

    ERIC Educational Resources Information Center

    Gilles, Carol; Dickinson, Jean

    1999-01-01

    Outlines the difference between "drilling for skills" and "becoming skillful readers" and shows how teachers can help students become skillful readers, writers, discussants, and thinkers through literature discussions. Discusses establishing a context for literature study, and initiating and maintaining literature study groups. (SR)

  14. Teacher Work Samples & Struggling Readers: Impacting Student Performance & Candidate Dispositions.

    ERIC Educational Resources Information Center

    Cartwright, Dennis D.; Blacklock, Karen K.

    This study reports positive impacts of a modified version of the teacher work sample on performance of struggling K-12 readers and on the dispositions of interns who worked with them. The participants included 55 university students in a year-long, senior-level internship and 55 elementary students identified as struggling readers. The…

  15. Tadpole Tales and Other Totally Terrific Treats for Readers Theatre.

    ERIC Educational Resources Information Center

    Fredericks, Anthony D.

    Readers theater is a storytelling device that stimulates the imagination and promotes all of the language arts. Readers theater is an oral interpretation of a piece of literature read in a dramatic style. It involves and motivates students, energizes the language arts program, and stimulates learning. Geared for children in grades 1-4, this book…

  16. Identifying What Works for Struggling Readers. Educator's Guide

    ERIC Educational Resources Information Center

    Slavin, Robert E.; Lake, Cynthia; Davis, Susan; Madden, Nancy A.

    2010-01-01

    This guide summarizes "Effective Programs for Struggling Readers: A Best Evidence Synthesis," a research review conducted by Johns Hopkins University's Center for Research and Reform in Education. The purpose of the review was to evaluate the achievement outcomes of alternative approaches for struggling readers in grade K-5: (1) one-to-one…

  17. Can Bold Typeface Improve Readers' Comprehension and Metacomprehension of Negation?

    ERIC Educational Resources Information Center

    Margolin, Sara J.

    2013-01-01

    Recent research on negation has demonstrated that while readers are aware that this text construction is difficult, they seem to be able to do little to improve their comprehension. The present research evaluated whether a change in typeface could improve comprehension and metacomprehension of negation. Results indicated that while readers were…

  18. Investigating the Relationship between Connectives and Readers' Reading Comprehension Level

    ERIC Educational Resources Information Center

    Gençer, Yusuf; ÇetiInkaya, Gökhan

    2015-01-01

    The purpose of this study is to investigate the relationship between connectives in Turkish texts and readers' reading comprehension. Research was conducted with a total of 50 teachers. In the study group, readers' reading comprehension was determined through 10 descriptive texts by using open-ended questions. The results of the analysis revealed…

  19. Writing for New Readers: Selections from a Writers' Manual.

    ERIC Educational Resources Information Center

    Van Horne, Marion

    This guide to writing material that will be read by new readers focuses on making the material readable, relevant, and effective. The first section of the guide gives an overview of this process, and addresses the following aspects of writing for new readers: (1) determining the audience and its interests; (2) elements of effective written…

  20. Addressing Learning Disabilities with UDL and Technology: Strategic Reader

    ERIC Educational Resources Information Center

    Hall, Tracey E.; Cohen, Nicole; Vue, Ge; Ganley, Patricia

    2015-01-01

    CAST created "Strategic Reader," a technology-based system blending Universal Design for Learning (UDL) and Curriculum-Based Measurement (CBM) in a digital learning environment to improve reading comprehension instruction. This experimental study evaluates the effectiveness of Strategic Reader using two treatment conditions for measuring…

  1. Stance, Navigation, and Reader Response in Expository Hypertext

    ERIC Educational Resources Information Center

    McEneaney, John E.; Li, Ledong; Allen, Kris; Guzniczak, Lizabeth

    2009-01-01

    This article reports on two studies investigating reader stance, navigation, and response in expository hypertext. Subjects in the studies included 69 and 147 adult readers prompted to adopt either an efferent or aesthetic stance when reading a 36-node expository hypertext. Reading was followed by recall and essay writing tasks. Results of the…

  2. A Comparative Review of Stratification Texts and Readers

    ERIC Educational Resources Information Center

    Peoples, Clayton D.

    2012-01-01

    Social stratification is a core substantive area within sociology. There are a number of textbooks and readers available on the market that deal with this central topic. In this article, I conduct a comparative review of (a) four stratification textbooks and (b) four stratification readers. (Contains 2 tables.)

  3. Cognitive Patterns of "Retarded" and Below-Average Readers.

    ERIC Educational Resources Information Center

    Leong, Che K.

    1980-01-01

    The cognitive patterns of 58 "retarded" and 38 below-average readers were compared with controls, according to Luria's simultaneous and successive modes of information processing. Factor Analysis showed different cognitive patterns for disabled and nondisabled readers. Reading skills, rather than cognitive ability, were shown to be…

  4. Benefits of Computer-Presented Speed Training for Poor Readers

    ERIC Educational Resources Information Center

    Irausquin, Rosemarie S.; Drent, Jeanine; Verhoeven, Ludo

    2005-01-01

    The effects of computer-presented automatization exercises in a group of 14 poor readers were assessed in comparison to a matched control group of 14 poor readers that received computer-presented exercises aimed at the use of context for word identification and comprehension. Training took place three sessions a week for 15 minutes per session and…

  5. Library Services to Distance Learners in the Commonwealth: A Reader.

    ERIC Educational Resources Information Center

    Watson, Elizabeth F., Ed.; Jagannathan, Neela, Ed.

    The provision of good library services is a crucial factor in determining the quality of distance education. This collection of articles acquaints readers with distance librarianship as it is practiced in developed and developing countries throughout the British Commonwealth. The reader includes: "Introduction" (Michael Wooliscroft);…

  6. Eye Movements Reveal Readers' Lexical Quality and Reading Experience

    ERIC Educational Resources Information Center

    Taylor, Jessica Nelson; Perfetti, Charles A.

    2016-01-01

    Two experiments demonstrate that individual differences among normal adult readers, including lexical quality, are expressed in silent reading at the word level. In the first of two studies we identified major dimensions of variability among college readers and among words using factor analysis. We then examined the effects of these dimensions of…

  7. Sexism Springs Eternal--in the Reader's Digest.

    ERIC Educational Resources Information Center

    Zimbardo, Philip G.; Meadow, Wendy

    This document reports on an empirical investigation of anti-women humor appearing in the Reader's Digest over three decades, revealing the operation of an unconscious sexist ideology. A systematic analysis was made of 1,069 jokes appearing in two featured columns of the Reader's Digest for the two-year periods 1947-48, 1957-58, and 1967-68.…

  8. Helping the Slow-Reader in the Primary School Classroom.

    ERIC Educational Resources Information Center

    Burke, Edmund V.

    1988-01-01

    A strategy is proposed for primary teachers to use with children who have reading difficulties. It includes developing a profile of each slow reader, testing each slow reader to determine reading level and specific weaknesses, and using certain practical methods to help solve the problem. (two references) (LB)

  9. Peace Pilgrim: A Readers Theatre Approach to Peace Education.

    ERIC Educational Resources Information Center

    Curran, Joanne M.

    Readers theater is a dramatic art that directs attention to the words of a text. During a performance, a reader (rather than an actor) presents and interprets the work within a narrative context intended by the author. In this project, the text, a collection of the words and writings of a woman known as the Peace Pilgrim, tells the story of her…

  10. Social Problem Fiction; A Source of Help for Retarded Readers.

    ERIC Educational Resources Information Center

    Iowa Univ., Iowa City. Special Education Curriculum Development Center.

    An annotated listing of over 500 major fiction books and basal readers provides suggestions for their use in library instruction, independent reading, class and small group discussion, and correlation with unit work and basal reader programs. The books are listed according to primary, intermediate, and advanced reading levels and books to be read…

  11. Who's Really Struggling?: Middle School Teachers' Perceptions of Struggling Readers

    ERIC Educational Resources Information Center

    Moreau, Leah K.

    2014-01-01

    This study explored middle school teachers' perceptions of struggling readers, including influences such as: understandings of components and factors relating to reading difficulties; views of struggling readers' behaviours and affect; classroom implications of their difficulties; and feelings of both competency and responsibility in the teaching…

  12. University Level Second Language Readers' Online Reading and Comprehension Strategies

    ERIC Educational Resources Information Center

    Park, Jaehan; Yang, Jaeseok; Hsieh, Yi Chin

    2014-01-01

    With the growing prevalence of Web 2.0 technologies and use of online resources in their classrooms, language learners have increasing exposure to online texts. In this study we attempted to understand how university level second language (L2) readers construct meaning when reading online. We investigated L2 readers' information-seeking strategies…

  13. Readers Theatre Handbook: A Dramatic Approach to Literature. Revised Edition.

    ERIC Educational Resources Information Center

    Coger, Leslie Irene; White, Melvin R.

    This handbook defines Readers Theatre, explores the selection and adaptation of material, and discusses casting, rehearsing, and performing of literature. It is divided into three major parts. Part One, "Origins, Concepts, and Precepts," emphasizes current trends in Readers Theatre and considers problems stemming from the introduction of…

  14. Infographics for Educational Purposes: Their Structure, Properties and Reader Approaches

    ERIC Educational Resources Information Center

    Yildirim, Serkan

    2016-01-01

    Infographics are one of the new educational environments used to provide information to their readers in a visual way. Infographics are designed to provide information to their readers using various visuals such as texts, pictures, drawings, diagrams, graphs, etc. The use of infographics becomes increasingly widespread both in advertising…

  15. Perceptions about "Good Readers": Analyzing Curricular, Linguistic, and Socioeconomic Differences

    ERIC Educational Resources Information Center

    Moses, Lindsey

    2015-01-01

    Students' perceptions about "what good readers do" are representative of their definition of reading and what they believe they should be striving toward as a reader. These beliefs about reading influence their motivation and ways they engage with texts. In this study, interviews were conducted with students in three first-grade…

  16. Teaching Materials for German. G3: Readers. Teaching Materials Lists.

    ERIC Educational Resources Information Center

    Centre for Information on Language Teaching, London (England).

    This publication, part of a bibliography of language teaching materials, lists 91 readers. It contains a contents list, annotated entries, an index of authors, editors, compilers and adapters, and a title index. In addition to basic bibliographical details, the following are provided for each entry: an annotation describing the reader, a statement…

  17. Reader Response Criticism in Children's Literature. Occasional Papers, 15.

    ERIC Educational Resources Information Center

    Benton, Michael

    Since the 1950s, literary study has experienced a major paradigm shift. M. H. Abrams' notion that the author, reader, and the signified world are arranged like satellites around a stable text has been succeeded by one that acknowledges the reader to be a central determinant in any "reading" of a text. Five themes related to reader…

  18. Choosing and Using Books with Adult New Readers.

    ERIC Educational Resources Information Center

    Weibel, Marguerite Crowley

    Written for teachers of adult literacy students and administrators of literacy programs, this book discusses a multitude of ways to use books appropriate for adult new readers. The book's first six chapters provide annotated lists of 639 books suitable for new readers, including titles, authors, dates, publishers, and suggested reading levels.…

  19. "Science in Society, Omnibus Pack, Readers A-L."

    ERIC Educational Resources Information Center

    Association for Science Education, Cambridge (England).

    Twelve readers have been written for use in a course developed by the Science in Society Project. Each reader (corresponding to a specific unit in the course) contains a number of short articles contributed by prominent figures in industry, the professions, the academic world, and in politics. Detailed suggestions on how to take advantage of the…

  20. Accelerated Reader[TM]. What Works Clearinghouse Intervention Report

    ERIC Educational Resources Information Center

    What Works Clearinghouse, 2010

    2010-01-01

    "Accelerated Reader"[TM] is a guided reading intervention used to supplement regular reading instruction in K-12 classrooms. Its aim is to improve students' reading skills through reading practice and quizzes on the books students read. The "Accelerated Reader"[TM] program calls for students to select and read a book and then…

  1. Designating Reader Perspective to Increase Comprehension and Interest

    ERIC Educational Resources Information Center

    Ramsay, Crystal M.; Sperling, Rayne A.

    2010-01-01

    In three experiments we examined whether reader perspective on a long expository text could be manipulated such that increased text interest and enhancement of two comprehension outcomes would result. In Experiment 1 we verified the viability of a new text for experimental purposes. We then assigned readers a perspective before reading in…

  2. Miscue Analysis: A Transformative Tool for Researchers, Teachers, and Readers

    ERIC Educational Resources Information Center

    Goodman, Yetta M.

    2015-01-01

    When a reader produces a response to a written text (the observed response) that is not expected by the listener, the result is called a miscue. Using psychosociolingustic analyses of miscues in the context of an authentic text, miscue analysis provides evidence to discover how readers read. I present miscue analysis history and development and…

  3. How Think-and-Feel-Aloud Instruction Influences Poetry Readers

    ERIC Educational Resources Information Center

    Eva-Wood, Amy L.

    2004-01-01

    Assuming readers' emotional responses can inform literary analysis, this study of poetry readers featured an instructional intervention that involved modeling both cognitive and affective reading processes through a think-and-feel-aloud pedagogy. Eleventh-grade students in 2 conditions participated in a 4-week unit on reading poetry. Control group…

  4. Breaking the Silence: Toward Improving LGBTQ Representation in Composition Readers

    ERIC Educational Resources Information Center

    Hudson, John

    2014-01-01

    Lesbian-Gay-Bisexual-Transgender-Queer (LGBTQ) representation in composition readers remains limited and is frequently nonexistent. In addition, the LGBTQ-related materials that do find their way into composition readers are often problematic. In this essay I explain why WPAs and composition teachers should be concerned about LGBTQ representation…

  5. Homophone Effects in Deaf Readers: Evidence from Lexical Decision

    ERIC Educational Resources Information Center

    Friesen, Deanna C.; Joanisse, Marc F.

    2012-01-01

    The current study examined the nature of deaf readers' phonological processing during online word recognition, and how this compares to similar effects in hearing individuals. Unlike many previous studies on phonological activation, we examined whether deaf readers activated phonological representations for words as opposed to pseudohomophones.…

  6. Chinese Deaf Readers Have Early Access to Parafoveal Semantics

    ERIC Educational Resources Information Center

    Yan, Ming; Pan, Jinger; Bélanger, Nathalie N.; Shu, Hua

    2015-01-01

    In the present study, we manipulated different types of information available in the parafovea during the reading of Chinese sentences and examined how deaf readers make use of the parafoveal information. Results clearly indicate that although the reading-level matched hearing readers make greater use of orthographic information in the parafovea,…

  7. An Unforgiving Enemy: AIDS. Student Reader and Teacher's Guide.

    ERIC Educational Resources Information Center

    Adult Education Services, Johnstown, PA.

    These adult basic education instructional materials on the prevention of Acquired Immune Deficiency Syndrome (AIDS) include a student reader and a teacher's guide. The student reader contains six chapters. Chapter 1 introduces two characters--Ricco and Francis--and focuses on the definition of AIDS, the three stages of the disease, the cause, and…

  8. ANNOUNCEMENT: To the readers of Kvantovaya Elektronika

    NASA Astrophysics Data System (ADS)

    1994-06-01

    The Radio i Svyaz' Press is preparing to publish several books on a number of topics in quantum electronics. These books are described below. They are included in the Federal Target Programme for the publication of books in Russia in 1994. In accordance with the Programme, the books are supported financially by the State to pay for the cost of paper and printing. Since at present the question of finance is very problematic and the actual cost of each of these books can be between 12 and 20 thousand roubles per copy, if the number of copies printed is 800-1500 the retail price of each copy would be between 15 and 25 thousand roubles. Consequently, if for any reason we do not receive financial support from the Federal Programme or if we do not receive guaranteed orders from readers, bearing in mind the real retail price, the Press will be unable to publish these books. Guaranteed orders are accepted from organisations and from individuals. The price includes postage on a cash-on-delivery basis if there is a guaranteed print order of 800-1500 copies of each book.

  9. Handheld E-Book Readers and Scholarship Report and Reader Survey: ACLS Humanities E-Book. White Paper No. 3

    ERIC Educational Resources Information Center

    Gielen, Nina

    2010-01-01

    This report describes a conversion experiment and subsequent reader survey conducted by the American Council of Learned Societies (ACLS) Humanities E-Book (HEB) in late 2009 and early 2010 to assess the viability of using scholarly monographs with handheld e-readers. As sample content, HEB selected six titles from its own online collection, three…

  10. Present-Day Microplate Tectonics of Tibet and its Relation to Rheological Stratification and Flow in the Lithosphere

    NASA Astrophysics Data System (ADS)

    Thatcher, W.

    2005-12-01

    deformation calculated from dynamic models for Tibet. These studies, based on continuum model assumptions, have concluded that fast S and strain axis orientations agree, suggesting coherent, coupled deformation through most of the thickness of the lithosphere. Comparison of fast S orientations and relative motions derived from the GPS microplate model suggest another interpretation. First, fast S orientations within identified microplates are nearly parallel to predicted block motions relative to Eurasia (rms misfit=14 deg.). Furthermore, fast S orientations across major strike-slip faults (Jiali, Kunlun) are parallel to predicted relative motions across block boundaries. Together these correlations suggest present-day microplate motions drive or resist more continuous ductile flow at depth. Because of the limited resolution of the S split observations, the depth at which block-like motion yields to ~continuum flow is poorly constrained. However, the source of splitting is unlikely to be in the asthenosphere, because correcting Tibet-Eurasia block motions to a hotspot reference frame (HS3-NUVEL1A) significantly degrades the match of `absolute' block motion to fast S orientations (rms misfit=40 deg.). The 1-2 sec magnitude of SKS split times seems to require significant flow in the mantle lithosphere, so ~rigid block motion of the crust could be directly coupled to upper mantle flow. However, crustal anisotropy may also be important, and low seismic velocity zones in the crust may mimic lower viscosity, so coupling between upper crustal blocks and ductile deformation of mantle lithosphere could occur via lower crustal flow.

  11. Productivity, Syntax, and Ideation in the Written Expression of Remedial and Achieving Readers.

    ERIC Educational Resources Information Center

    Anderson, Peggy L.

    1989-01-01

    The study compared the written expression of 56 fourth to fifth grade remedial readers and achieving readers. Significant differences favoring the achieving readers were found in three areas: productivity, syntax, and level of ideation. Remedial readers tended toward the same kinds of errors as achieving readers but with an increased frequency.…

  12. Inter- and Intra-Assay Reproducibility of Microplate Alamar Blue Assay Results for Isoniazid, Rifampicin, Ethambutol, Streptomycin, Ciprofloxacin, and Capreomycin Drug Susceptibility Testing of Mycobacterium tuberculosis▿ †

    PubMed Central

    Leonard, Brian; Coronel, Jorge; Siedner, Mark; Grandjean, Louis; Caviedes, Luz; Navarro, Pilar; Gilman, Robert H.; Moore, David A. J.

    2008-01-01

    The intersample and intrasample variability of the results obtained with the microplate Alamar blue assay for the indirect drug susceptibility testing of Mycobacterium tuberculosis was investigated. Between 1.2 and 8.5% of paired MICs differed by more than one twofold dilution, resulting in discordant susceptible-resistant designations at frequencies between 0.6% (rifampin) and 18.9% (ethambutol). PMID:18701659

  13. Clarithromycin Susceptibility Testing of Mycobacterium avium Complex Using 2,3-Diphenyl-5-thienyl-(2)-tetrazolium Chloride Microplate Assay with Middlebrook 7H9 Broth

    PubMed Central

    Park, Young Kil; Koh, Won-Jung; Kim, Shin Ok; Shin, Sonya; Kim, Bum Joon; Cho, Sang-Nae; Lee, Sun Min

    2009-01-01

    A series of 119 Mycobacterium avium complex isolates were subjected to clarithromycin susceptibility testing using microplates containing 2,3-diphenyl-5-thienyl-(2)-tetrazolium chloride (STC). Among 119 isolates, 114 (95.8%) were susceptible to clarithromycin and 5 were resistant according to the new and the standard method. STC counts the low cost and reduces the number of procedures needed for susceptibility testing. PMID:19543518

  14. Quantitation of biotin-binding immunoglobulins G, A, and M in Human Sera Using F(ab')2anti-human immunoglobulin-coated microplates.

    PubMed

    Muratsugu, Makoto; Yazawa, Ayaka; Fujiwara, Sami; Nishida, Satsuki; Fukui, Toru

    2008-03-01

    Biotin-binding IgG (B-IgG) in human sera was quantified using previously developed F(ab')(2)anti-human IgG-coated multiwell microplates (Muratsugu M. et al., 2003, Biol. Pharm. Bull., 26, 1605--1608). The levels of B-IgG in sera, however, were higher than those we predicted. In this study, we modified the assay using F(ab')2anti-human IgG-coated multiwell microplates and successfully quantified the levels of B-IgG in sera. The cause of the unpredicted results was discussed in the text. In addition, the levels of biotin-binding IgA (B-IgA) and IgM (B-IgM) in sera could be measured using F(ab')2anti-human IgA- or IgM-coated multiwell microplates. We quantified B-IgG, B-IgA, and B-IgM in sera from healthy specimens and patients with bronchial asthma, atopic dermatitis, epilepsy, and juvenile rheumatoid arthritis.

  15. RFID reader design for identification of industrial metallic objects

    NASA Astrophysics Data System (ADS)

    Szczurkowski, M.; Jankowski, H.; Worek, C.; Maksymowicz, L. J.

    2006-10-01

    An electronic circuit design of the RFID (Radio Frequency Identification) reader was the aim of our research. This device is designed for identification of transponders, which are present on the surface of metal elements or inside machine elements made of metal. In order to achieve that, we optimized the antenna reader circuit, that works in near field (magnetic coupling), by means of field flux lines analysis (using finite elements method) and experimental data collecting. Finally, one obtaines a specific shaped magnetic field around the reader.

  16. Usability and Accessibility of eBay by Screen Reader

    NASA Astrophysics Data System (ADS)

    Buzzi, Maria Claudia; Buzzi, Marina; Leporini, Barbara; Akhter, Fahim

    The evolution of Information and Communication Technology and the rapid growth of the Internet have fuelled a great diffusion of eCommerce websites. Usually these sites have complex layouts crowded with active elements, and thus are difficult to navigate via screen reader. Interactive environments should be properly designed and delivered to everyone, including the blind, who usually use screen readers to interact with their computers. In this paper we investigate the interaction of blind users with eBay, a popular eCommerce website, and discuss how using the W3C Accessible Rich Internet Applications (WAI-ARIA) suite could improve the user experience when navigating via screen reader.

  17. Detection of c-reactive protein based on a magnetic immunoassay by using functional magnetic and fluorescent nanoparticles in microplates.

    PubMed

    Yang, S F; Gao, B Z; Tsai, H Y; Fuh, C Bor

    2014-11-07

    We report the preparation and application of biofunctional nanoparticles to detect C-reactive protein (CRP) in magnetic microplates. A CRP model biomarker was used to test the proposed detection method. Biofunctional magnetic nanoparticles, CRP, and biofunctional fluorescent nanoparticles were used in a sandwich nanoparticle immunoassay. The CRP concentrations in the samples were deduced from the reference plot, using the fluorescence intensity of the sandwich nanoparticle immunoassay. When biofunctional nanoparticles were used to detect CRP, the detection limit was 1.0 ng ml(-1) and the linear range was between 1.18 ng ml(-1) and 11.8 μg ml(-1). The results revealed that the method involving biofunctional nanoparticles exhibited a lower detection limit and a wider linear range than those of the enzyme-linked immunosorbent assay (ELISA) and most other methods. For CRP measurements of serum samples, the differences between this method and ELISA in CRP measurements of serum samples were less than 13%. The proposed method can reduce the analysis time to one-third that of ELISA. This method demonstrates the potential to replace ELISA for rapidly detecting biomarkers with a low detection limit and a wide dynamic range.

  18. Late Pleistocene and Holocene uplift history of Cyprus: implications for active tectonics along the southern margin of the Anatolian microplate

    USGS Publications Warehouse

    Harrison, R.W.; Tsiolakis, E.; Stone, B.D.; Lord, A.; McGeehin, J.P.; Mahan, S.A.; Chirico, P.

    2013-01-01

    The nature of the southern margin of the Anatolian microplate during the Neogene is complex, controversial and fundamental in understanding active plate-margin tectonics and natural hazards in the Eastern Mediterranean region. Our investigation provides new insights into the Late Pleistocene uplift history of Cyprus and the Troodos Ophiolite. We provide isotopic (14C) and radiogenic (luminescence) dates of outcropping marine sediments in eastern Cyprus that identify periods of deposition during marine isotope stages (MIS) 3, 4, 5 and 6. Past sea-levels indicated by these deposits are c. 95±25 m higher in elevation than estimates of worldwide eustatic sea-level. An uplift rate of c. 1.8 mm/year and possibly as much as c. 4.1 mm/year in the past c. 26–40 ka is indicated. Holocene marine deposits also occur at elevations higher than those expected for past SL and suggest uplift rates of c. 1.2–2.1 mm/year. MIS-3 marine deposits that crop out in southern and western Cyprus indicate uniform island-wide uplift. We propose a model of tectonic wedging at a plate-bounding restraining bend as a mechanism for Late Pleistocene to Holocene uplift of Cyprus; uplift is accommodated by deformation and seismicity along the margins of the Troodos Ophiolite and re-activation of its low-angle, basal shear zone.

  19. Enhanced Microwave Absorption Properties of Flexible Polymer Composite Based on Hexagonal NiCo2O4 Microplates and PVDF

    NASA Astrophysics Data System (ADS)

    Luo, Hui; Wang, Xian; Song, Kai; Yang, Jing; Gong, Rongzhou

    2016-08-01

    Hexagonal NiCo2O4 microplates were synthesized via a facile one-pot hydrothermal method and followed by a subsequent annealing process. The complex permittivity and permeability of a NiCo2O4 and polyvinylidene fluoride (PVDF) composite were investigated over 2-18 GHz. The experiment indicated that the good microwave absorption performance of NiCo2O4@PVDF depends on dielectric loss and quarter-wavelength cancellation. Our results show that the absorption frequency bandwidth of reflection loss (RL) less than -20 dB for the NiCo2O4@PVDF composite can be measured over the frequency range of 3-15.5 GHz with an absorbing thickness that varies in the range of 1.25-5 mm. Furthermore, an optimal RL of -44.8 dB was observed at 10.7 GHz with a thickness of 1.75 mm. The loss mechanism is also discussed.

  20. [Validation of antibody screening by indirect antigloblin test and ABO blood typing by filtration and microplate techniques: assessment of robustness].

    PubMed

    Mannessier, L; Delamaire, M; Bouix, O; Krause, C; Roubinet, F

    2006-10-01

    According to requirements of the French Committee for Accreditation (Cofrac), it is essential to use validated and standardised methods in Immunohematology. This imposes first the knowledge of metrological tolerances for all the technics. Two multicenter studies were carried out to define the maximal acceptable deviations concerning incubation temperature and time, volumes of patient plasma and tests cells for antibody screening using indirect antiglobulin test on one hand and for reverse grouping on another hand. All equipment used (temperature test chamber, chronometer, pipettes) were calibrated according to Cofrac standards. The antibody screenings were performed manually using 3 different filtration systems: ID Diamed, Biovue Ortho and Scangel Biorad, the same tests cells, a standard 20 ng/mL anti RH1, a positive control anti KEL1 and a negative control; the reverse blood grouping was performed manually using the above mentionned filtration systems and microplate technic with the same A1 and B test cells. These two studies showed that all the tests from the multiples combinations of the above parameters gave the same results and allowed us to define a range of tolerance for 4 critical physical parameters involved in the antibody screening and blood typing.

  1. Susceptibility profiles of Nocardia spp. to antimicrobial and antituberculotic agents detected by a microplate Alamar Blue assay

    PubMed Central

    Zhao, Pan; Zhang, Xiujuan; Du, Pengcheng; Li, Guilian; Li, Luxi; Li, Zhenjun

    2017-01-01

    Nocardia species are ubiquitous in natural environments and can cause nocardiosis. Trimethoprim-sulfamethoxazole has long been the monotherapy treatment of choice, but resistance to this treatment has recently emerged. In this study, we used microplate Alamar Blue assays to determine the antimicrobial susceptibility patterns of 65 standard Nocardia isolates, including 28 type strains and 20 clinical Nocardia isolates, to 32 antimicrobial agents, including 13 little studied drugs. Susceptibility to the most commonly used drug, trimethoprim-sulfamethoxazole, was observed in 98% of the isolates. Linezolid, meropenem, and amikacin were also highly effective, with 98%, 95%, and 90% susceptibility, respectively, among the isolates. The isolates showed a high percentage of resistance or nonsusceptibility to isoniazid, rifampicin, and ethambutol. For the remaining antimicrobials, resistance was species-specific among isolates and was observed in traditional drug pattern types. In addition, the antimicrobial susceptibility profiles of a variety of rarely encountered standard Nocardia species are reported, as are the results for rarely reported clinical antibiotics. We also provide a timely update of antimicrobial susceptibility patterns that includes three new drug pattern types. The data from this study provide information on antimicrobial activity against specific Nocardia species and yield important clues for the optimization of species-specific Nocardia therapies. PMID:28252662

  2. Microplate Assay of α-Glucosidase and Its Inhibitors Based on the Direct Reduction of Molybdosilicate by Glucose.

    PubMed

    Katano, Hajime; Takakuwa, Masahiro; Itoh, Takafumi; Hibi, Takao

    2015-01-01

    A colorimetric method for monosaccharide determination (Anal. Sci., 2013, 29, 1021) was optimized for the high-throughput screening of α-glucosidase, which hydrolyzes an α-1,4-glycosidic bond of starch and related oligo- and polysaccharides, followed by the release of D-glucose from the non-reducing ends. In a microplate, 40 μL of a sample solution was mixed with 160 μL of a 50 mM Na2SiO3, 600 mM Na2MoO4, 1.5 M CH3COOH, and 20% (v/v) dimethyl sulfoxide solution, which was yellowish due to the formation of a yellow molybdosilicate. The mixture was kept at 80°C for 60 min. In the mixture, glucose reduced the Mo(VI) species directly to form a blue heteropolymolybdate(V/VI). Thus, 0.1 mM level glucose can be determined by the color change from yellow to blue. Since maltose cannot render the mixture blue as strongly as glucose, the present method has been successfully applied to a microtiter plate assay of α-glucosidase with the disaccharide. Also, the method has been applied to an assay of α-glucosidase inhibitors, acarbose and quercetin.

  3. Reversal and Rotation Errors by Normal and Retarded Readers

    ERIC Educational Resources Information Center

    Black, F. William

    1973-01-01

    Reports an investigation of the incidence of and relationships among word and letter reversals in writing and Bender-Gestalt rotation errors in matched samples of normal and retarded readers. No significant diffenences were found in the two groups. (TO)

  4. How Readers and Advertisers Benefit from Local Newspaper Competition.

    ERIC Educational Resources Information Center

    Everett, Shu-Ling Chen; Everett, Stephen E.

    1989-01-01

    Explores relations among three competitive schemes with respect to newspapers' price structures, including advertising rates and prices to consumers. Finds that readers get some benefit from greater competition, but that advertisers do not. (MM)

  5. ERIC/RCS: Reader Response in the Classroom.

    ERIC Educational Resources Information Center

    Holbrook, Hilary Taylor

    1987-01-01

    Explores briefly the New Criticism that dominated literature instruction until recently and then provides an overview of reader response theory and how response approaches can be used in the classroom to enhance reading. (NKA)

  6. What Is That Lapping the Miles?: Responding to Reader Response.

    ERIC Educational Resources Information Center

    Seib, Kenneth

    1995-01-01

    Responds to an article in an earlier issue of this journal about using reading response in a college literature classroom. Argues that the use of reader-response theory with two-year college students requires some caution. (SR)

  7. Late Miocene to Recent formation of the Aure-Moresby fold-thrust belt and foreland basin as a consequence of Woodlark microplate rotation, Papua New Guinea

    NASA Astrophysics Data System (ADS)

    Ott, Bryan; Mann, Paul

    2015-06-01

    The Aure-Moresby fold-thrust belt and Aure-Moresby foreland basin are located in the eastern Gulf of Papua (GOP), Papua New Guinea (PNG), and formed during late Miocene-Recent as the result of large-scale, counterclockwise rotation of the 355,000 km2 Woodlark microplate. To document the structure, stratigraphy, and age of convergent deformation along the poorly studied, western edge of the rotating Woodlark microplate, we integrate results of 2,538 km of previously unpublished 2-D seismic reflection data with onshore geologic and GPS studies from previous workers. The late Miocene Aure-Moresby fold-thrust belt is a 400 km long, northwest-trending fold-belt exposed onshore in Papua New Guinea that plunges to the southeast, where continuous folds and northeast-dipping thrusts can be imaged in the subsurface for more than 250 km. The arcuate trend of the Aure-Moresby fold-thrust belt along the southwestern coast and offshore areas of the Papuan peninsula parallels the shape of the adjacent, offshore Aure-Moresby foreland basin and the strike of the transpressional segment of the left-lateral Owen-Stanley fault zone (OSFZ) passing along the center of the Papuan peninsula. As the OSFZ becomes more transtensional east of 148°E, folds of the Aure-Moresby fold-thrust belt along southern coast of the peninsula become less prominent, and the adjacent Aure-Moresby foreland basin transitions into an undeformed Cenozoic passive margin setting. These observations of convergent an left-lateral deformation along the Aure-Moresby fold-thrust belt are consistent with: (1) counterclockwise rotation of the Woodlark microplate known from regional GPS studies; (2) coeval opening of the Woodlark basin along its southern edge in the late Miocene; and (3) rapid subduction at the New Britain trench along its northern edge. The kinematics of the rotating Woodlark microplate are driven by slab pull forces acting on the actively subducting northern edge of the microplate.

  8. Flexible electronics enters the e-reader market

    NASA Astrophysics Data System (ADS)

    Banks, Michael

    2010-02-01

    A company that was spun off from the physics department at the University of Cambridge in the UK 10 years ago released its first product last month. Plastic Logic, founded by Henning Sirringhaus and Richard Friend, launched an electronic reader that can display books, magazines and newspapers on a flexible, lightweight plastic display. The reader commercializes pioneering work first started over 20 years ago at the lab by the two physicists, who are based in the department's optoelectronics group.

  9. An IoT Reader for Wireless Passive Electromagnetic Sensors.

    PubMed

    Galindo-Romera, Gabriel; Carnerero-Cano, Javier; Martínez-Martínez, José Juan; Herraiz-Martínez, Francisco Javier

    2017-03-28

    In the last years, many passive electromagnetic sensors have been reported. Some of these sensors are used for measuring harmful substances. Moreover, the response of these sensors is usually obtained with laboratory equipment. This approach highly increases the total cost and complexity of the sensing system. In this work, a novel low-cost and portable Internet-of-Things (IoT) reader for passive wireless electromagnetic sensors is proposed. The reader is used to interrogate the sensors within a short-range wireless link avoiding the direct contact with the substances under test. The IoT functionalities of the reader allows remote sensing from computers and handheld devices. For that purpose, the proposed design is based on four functional layers: the radiating layer, the RF interface, the IoT mini-computer and the power unit. In this paper a demonstrator of the proposed reader is designed and manufactured. The demonstrator shows, through the remote measurement of different substances, that the proposed system can estimate the dielectric permittivity. It has been demonstrated that a linear approximation with a small error can be extracted from the reader measurements. It is remarkable that the proposed reader can be used with other type of electromagnetic sensors, which transduce the magnitude variations in the frequency domain.

  10. Implementation and extension of a GNU-Radio RFID reader

    NASA Astrophysics Data System (ADS)

    Smietanka, G.; Brato, S.; Freudenberg, M.; Götze, J.

    2013-07-01

    The development of a flexible software defined RFID is discused. Commercial reader systems only allow a top level view on the communication and restrict the variation for many transmission parameters. Recently a software reader from the CGran project was proposed which uses the GNU Radio environment in combination with an USRP front end. Because most of the signal processing is done on a common host PC, this reader offers high flexibility, but also has several disadvantages. One of the main hardware limitations is the usage of only one separated antenna per transmit and receive path. Commercial readers usually use four antennas which are time multiplexed and can be used as transmitter and receiver. In this work a HF multiplexer for the USRP device is introduced. With this extension up to four transmit and receive antennas can be used in combination with the software reader. It is shown that the multiplexer achieves good read rates for a switching interval of 100 ms. Using this multiplexer the read range of the system decrease compared to the basic software reader, but distances over two meters can still be realized without additional hardware extensions.

  11. When High-Capacity Readers Slow Down and Low-Capacity Readers Speed Up: Working Memory and Locality Effects.

    PubMed

    Nicenboim, Bruno; Logačev, Pavel; Gattei, Carolina; Vasishth, Shravan

    2016-01-01

    We examined the effects of argument-head distance in SVO and SOV languages (Spanish and German), while taking into account readers' working memory capacity and controlling for expectation (Levy, 2008) and other factors. We predicted only locality effects, that is, a slowdown produced by increased dependency distance (Gibson, 2000; Lewis and Vasishth, 2005). Furthermore, we expected stronger locality effects for readers with low working memory capacity. Contrary to our predictions, low-capacity readers showed faster reading with increased distance, while high-capacity readers showed locality effects. We suggest that while the locality effects are compatible with memory-based explanations, the speedup of low-capacity readers can be explained by an increased probability of retrieval failure. We present a computational model based on ACT-R built under the previous assumptions, which is able to give a qualitative account for the present data and can be tested in future research. Our results suggest that in some cases, interpreting longer RTs as indexing increased processing difficulty and shorter RTs as facilitation may be too simplistic: The same increase in processing difficulty may lead to slowdowns in high-capacity readers and speedups in low-capacity ones. Ignoring individual level capacity differences when investigating locality effects may lead to misleading conclusions.

  12. When High-Capacity Readers Slow Down and Low-Capacity Readers Speed Up: Working Memory and Locality Effects

    PubMed Central

    Nicenboim, Bruno; Logačev, Pavel; Gattei, Carolina; Vasishth, Shravan

    2016-01-01

    We examined the effects of argument-head distance in SVO and SOV languages (Spanish and German), while taking into account readers' working memory capacity and controlling for expectation (Levy, 2008) and other factors. We predicted only locality effects, that is, a slowdown produced by increased dependency distance (Gibson, 2000; Lewis and Vasishth, 2005). Furthermore, we expected stronger locality effects for readers with low working memory capacity. Contrary to our predictions, low-capacity readers showed faster reading with increased distance, while high-capacity readers showed locality effects. We suggest that while the locality effects are compatible with memory-based explanations, the speedup of low-capacity readers can be explained by an increased probability of retrieval failure. We present a computational model based on ACT-R built under the previous assumptions, which is able to give a qualitative account for the present data and can be tested in future research. Our results suggest that in some cases, interpreting longer RTs as indexing increased processing difficulty and shorter RTs as facilitation may be too simplistic: The same increase in processing difficulty may lead to slowdowns in high-capacity readers and speedups in low-capacity ones. Ignoring individual level capacity differences when investigating locality effects may lead to misleading conclusions. PMID:27014113

  13. Reading skill components and impairments in middle school struggling readers

    PubMed Central

    Cirino, Paul T.; Romain, Melissa A.; Barth, Amy E.; Tolar, Tammy D.; Fletcher, Jack M.; Vaughn, Sharon

    2012-01-01

    This study investigated how measures of decoding, fluency, and comprehension in middle school students overlap with one another, whether the pattern of overlap differs between struggling and typical readers, and the relative frequency of different types of reading difficulties. The 1,748 sixth, seventh, and eighth grade students were oversampled for struggling readers (n = 1,025) on the basis of the state reading comprehension proficiency measure. Multigroup confirmatory factor analyses showed partial invariance among struggling and typical readers (with differential loadings for fluency and for comprehension), and strict invariance for decoding and a combined fluency/comprehension factor. Among these struggling readers, most (85 %) also had weaknesses on nationally standardized measures, particularly in comprehension; however, most of these also had difficulties in decoding or fluency. These results show that the number of students with a specific comprehension problem is lower than recent consensus reports estimate and that the relation of different reading components varies according to struggling versus proficient readers. PMID:24000271

  14. Reading between the lines: Societal norms in Sierra Leonean readers

    NASA Astrophysics Data System (ADS)

    Brams, Patricia

    1980-12-01

    A content analysis of primary school readers of Sierra Leone revealed that the particular modern and traditional normative aspirations expressed in the National Development Plan for 1974/75-1978/79 were also generally reflected in the children's readers. Compared to the 1964 readers, the first indigenous readers developed circa 1977 contained markedly greater emphasis on traditional norms, though modernity norms continued to dominate, and substantially less emphasis on Efficacy (a central aspect of modernity) and on Non-parochial Affiliation. This closely corresponded with the intent of the National Plan to continue on a modernizing course employing the traditional norms of Manual Labor and Social Cohesion in a grassroots effort to develop the agricultural sector, with nationalism in a less important role. Apart from lesser emphasis on Efficacy and underemphasis on the Value of Education which were discordant with national goals, the 1977 readers seem to provide children and teachers with a fairly accurate image of the national ethos. This may help to account for the generally positive relationship that has been found between schooling and economic development.

  15. A Novel Automatic Rapid Diagnostic Test Reader Platform

    PubMed Central

    Ozkan, Haydar; Kayhan, Osman Semih

    2016-01-01

    A novel automatic Rapid Diagnostic Test (RDT) reader platform is designed to analyze and diagnose target disease by using existing consumer cameras of a laptop-computer or a tablet. The RDT reader is useable with numerous lateral immunochromatographic assays and similar biomedical tests. The system has two different components, which are 3D-printed, low-cost, tiny, and compact stand and a decision program named RDT-AutoReader 2.0. The program takes the image of RDT, crops the region of interest (ROI), and extracts the features from the control end test lines to classify the results as invalid, positive, or negative. All related patient's personal information, image of ROI, and the e-report are digitally saved and transferred to the related clinician. Condition of the patient and the progress of the disease can be monitored by using the saved data. The reader platform has been tested by taking image from used cassette RDTs of rotavirus (RtV)/adenovirus (AdV) and lateral flow strip RDTs of Helicobacter pylori (H. pylori) before discarding them. The created RDT reader can also supply real-time statistics of various illnesses by using databases and Internet. This can help to inhibit propagation of contagious diseases and to increase readiness against epidemic diseases worldwide. PMID:27190549

  16. Alternate reading strategies and variable asymmetry of the planum temporale in adult resilient readers

    PubMed Central

    Welcome, Suzanne E.; Leonard, Christiana M.; Chiarello, Christine

    2010-01-01

    Resilient readers are characterized by impaired phonological processing despite skilled text comprehension. We investigated orthographic and semantic processing in resilient readers to examine mechanisms of compensation for poor phonological decoding. Performance on phonological (phoneme deletion, pseudoword reading), orthographic (orthographic choice, orthographic analogy), and semantic (semantic priming, homograph resolution) tasks was compared between resilient, poor and proficient readers. Asymmetry of the planum temporale was investigated in order to determine whether atypical readers showed unusual morphology in this language-relevant region. Resilient readers showed deficits on phonological tasks similar to those shown by poor readers. We obtained no evidence that resilient readers compensate via superior orthographic processing, as they showed neither exceptional orthographic skill nor increased reliance on orthography to guide pronunciation. Resilient readers benefited more than poor or proficient readers from semantic relationships between words and experienced greater difficulty when such relationships were not present. We suggest, therefore, that resilient readers compensate for poor phonological decoding via greater reliance on word meaning relationships. The reading groups did not differ in mean asymmetry of the planum temporale. However, resilient readers showed greater variability in planar asymmetry than proficient readers. Poor readers also showed a trend towards greater variability in planar asymmetry, with more poor readers than proficient readers showing extreme asymmetry. Such increased variability suggests that university students with less reading skill display less well regulated brain anatomy than proficient readers. PMID:20223512

  17. Changing Perspectives: The Implied Reader in Australian Children's Literature, 1841-1994.

    ERIC Educational Resources Information Center

    Saxby, Maurice

    1995-01-01

    Looks back at 150 years of writing for children in Australia through the illumination cast by Wolfgang Iser's notion of "the implied reader." Suggests that the implied reader in 1841 was of the educated elite; by 1941 the reader was more likely to be middle-class with popular tastes and interests; and the reader of 1991 reflected a…

  18. Using Twitter to Teach Reader-Oriented Biblical Interpretation: "Tweading" the Gospel of Mark

    ERIC Educational Resources Information Center

    Williamson, Robert, Jr.

    2013-01-01

    Twitter offers an engaging way to introduce students to reader-oriented interpretation of the Bible. The exercise described here introduces students to the idea that the reader has a role in the production of a text's meaning, which thus varies from reader to reader. Twitter enables us to capture the real-time thoughts of a variety of…

  19. Implication versus Inference: Analyzing Writer and Reader Representations in Business Texts

    ERIC Educational Resources Information Center

    Jameson, Daphne A.

    2004-01-01

    The narrative concepts of the implied author and implied reader elucidate how business texts represent writers and readers. It is important, though, to distinguish carefully between writers' "implications" and readers' "inferences." Instructors should contrast implied versus inferred writers and readers, provide multiple ways to comprehend these…

  20. Comparative cytotoxic and genotoxic potential of 13 drinking water disinfection by-products using a microplate-based cytotoxicity assay and a developed SOS/umu assay.

    PubMed

    Zhang, Shao-Hui; Miao, Dong-Yue; Tan, Li; Liu, Ai-Lin; Lu, Wen-Qing

    2016-01-01

    The implications of disinfection by-products (DBPs) present in drinking water are of public health concern because of their potential mutagenic, carcinogenic and other toxic effects on humans. In this study, we selected 13 main DBPs found in drinking water to quantitatively analyse their cytotoxicity and genotoxicity using a microplate-based cytotoxicity assay and a developed SOS/umu assay in Salmonella typhimurium TA1535/pSK1002. With the developed SOS/umu test, eight DBPs: 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-fura3-chloro-4-(dichloromethyl)-5-hydroxy-2-[5H]-furanone (MX), dibromoacetonitrile (DBN), iodoacetic acid (IA), bromochloroacetonitrile (BCN), bromoacetic acid (BA), trichloroacetonitrile (TCN), dibromoacetic acid (DBA) and dichloroacetic acid (DCA) were significantly genotoxic to S. typhimurium. Three DBPs: chloroacetic acid (CA), trichloroacetic acid (TCA) and dichloroacetonitrile (DCN) were weakly genotoxic, whereas the remaining DBPs: chloroacetonitrile (CN) and chloral hydrate (CH) were negative. The rank order in decreasing genotoxicity was as follows: MX > DBN > IA > BCN > BA > TCN > DBA > DCA > CA, TCA, DCN > CN, CH. MX was approximately 370 000 times more genotoxic than DCA. In the microplate-based cytotoxicity assay, cytotoxic potencies of the 13 DBPs were compared and ranked in decreasing order as follows: MX > IA > DBN > BCN > BA > TCN > DCN > CA > DCA > DBA > CN > TCA > CH. MX was approximately 19 200 times more cytotoxic than CH. A statistically significant correlation was found between cytotoxicity and genotoxicity of the 13 DBPs in S. typhimurium. Results suggest that microplate-based cytotoxicity assay and the developed SOS/umu assay are feasible tools for analysing the cytotoxicity and genotoxicity of DBPs, particularly for comparing their toxic intensities quantitatively.

  1. Development, Optimization, and Validation of a Microplate Bioassay for Relative Potency Determination of Linezolid Using a Design Space Concept, and its Measurement Uncertainty.

    PubMed

    Saviano, Alessandro Morais; Francisco, Fabiane Lacerda; Ostronoff, Celina Silva; Lourenço, Felipe Rebello

    2015-01-01

    The aim of this study was to develop, optimize, and validate a microplate bioassay for relative potency determination of linezolid in pharmaceutical samples using quality-by-design and design space approaches. In addition, a procedure is described for estimating relative potency uncertainty based on microbiological response variability. The influence of culture media composition was studied using a factorial design and a central composite design was adopted to study the influence of inoculum proportion and triphenyltetrazolium chloride in microbial growth. The microplate bioassay was optimized regarding the responses of low, medium, and high doses of linezolid, negative and positive controls, and the slope, intercept, and correlation coefficient of dose-response curves. According to optimization results, design space ranges were established using: (a) low (1.0 μg/mL), medium (2.0 μg/mL), and high (4.0 μg/mL) doses of pharmaceutical samples and linezolid chemical reference substance; (b) Staphylococcus aureus ATCC 653 in an inoculum proportion of 10%; (c) antibiotic No. 3 culture medium pH 7.0±0.1; (d) 6 h incubation at 37.0±0.1ºC; and (e) addition of 50 μL of 0.5% (w/v) triphenyltetrazolium chloride solution. The microplate bioassay was linear (r2=0.992), specific, precise (repeatability RSD=2.3% and intermediate precision RSD=4.3%), accurate (mean recovery=101.4%), and robust. The overall measurement uncertainty was reasonable considering the increased variability inherent in microbiological response. Final uncertainty was comparable with those obtained with other microbiological assays, as well as chemical methods.

  2. Interaction of the Koinobiont Parasitoid Microplitis rufiventris of the Cotton Leafworm, Spodoptera littoralis, with Two Entomopathogenic Rhabditids, Heterorhabditis bacteriophora and Steinernema carpocapsae

    PubMed Central

    Atwa, Atwa A.; Hegazi, Esmat M.; Khafagi, Wedad E.; El-Aziz, Gehan M. Abd

    2013-01-01

    Entomopathogenic nematodes are generally considered beneficial organisms. However, they can affect beneficial insects such as parasitoids. The interaction between the entomopathogenic nematodes Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae) and Steinernema carpocapsae Weiser, and the parasitoid Microplitis rufiventris Kokujev (Hymenoptera: Braconidae) was investigated in the laboratory. In non-parasitized hosts, Spodoptera littoralis Boisduval (Lepidoptera: Noctuidae) larvae exposed to H. bacteriophora showed a higher percent mortality than those exposed to S. carpocapsae. Both nematodes were able to invade and propagate in non-parasitized S. littoralis larvae and those parasitized by M. rufiventris. Both nematode species reproduced in Microplitis-parasitized hosts, but there was a higher number of nematodes in non-parasitized larvae. S. carpocapsae yielded higher numbers of infective juveniles than H. bacteriophora. Generally, the number of nematodes harvested increased as their host's size increased. The interaction between the nematodes and parasitoid favored the nematodes when the nematodes were inoculated during the parasitoid egg stage or the young parasitoid larvae, thus giving the nematodes a better chance to grow and reproduce, resulting in the death of the parasitoid larvae. Conversely, when the nematodes were inoculated during the late larval instar of the parasitoid, the competition partially favored the wasp, thus giving approximately 50% of the wasps a better chance to develop, emerge, and reproduce, providing evidence that both nematodes and wasps could reproduce in the same host. Egg maturation of female wasps derived from nematode-infected hosts was not significantly different than those from control hosts. The combined application of nematodes and parasitoids may be beneficial if the detrimental effects of the nematodes on the parasitoid could be avoided by precisely timing the application strategies. It is clear that

  3. Optical benchmarking of security document readers for automated border control

    NASA Astrophysics Data System (ADS)

    Valentín, Kristián.; Wild, Peter; Å tolc, Svorad; Daubner, Franz; Clabian, Markus

    2016-10-01

    Authentication and optical verification of travel documents upon crossing borders is of utmost importance for national security. Understanding the workflow and different approaches to ICAO 9303 travel document scanning in passport readers, as well as highlighting normalization issues and designing new methods to achieve better harmonization across inspection devices are key steps for the development of more effective and efficient next- generation passport inspection. This paper presents a survey of state-of-the-art document inspection systems, showcasing results of a document reader challenge investigating 9 devices with regards to optical characteristics.

  4. Hand-held portable microarray reader for biodetection

    DOEpatents

    Thompson, Deanna Lynn; Coleman, Matthew A; Lane, Stephen M; Matthews, Dennis L; Albala, Joanna; Wachsmann-Hogiu, Sebastian

    2013-04-23

    A hand-held portable microarray reader for biodetection includes a microarray reader engineered to be small enough for portable applications. The invention includes a high-powered light-emitting diode that emits excitation light, an excitation filter positioned to receive the excitation light, a slide, a slide holder assembly for positioning the slide to receive the excitation light from the excitation filter, an emission filter positioned to receive the excitation light from the slide, a lens positioned to receive the excitation light from the emission filter, and a CCD camera positioned to receive the excitation light from the lens.

  5. Rapid Detection of an ABT-737-Sensitive Primed for Death State in Cells Using Microplate-Based Respirometry

    PubMed Central

    Clerc, Pascaline; Carey, Gregory B.; Mehrabian, Zara; Wei, Michael; Hwang, Hyehyun; Girnun, Geoffrey D.; Chen, Hegang; Martin, Stuart S.; Polster, Brian M.

    2012-01-01

    Cells that exhibit an absolute dependence on the anti-apoptotic BCL-2 protein for survival are termed “primed for death” and are killed by the BCL-2 antagonist ABT-737. Many cancers exhibit a primed phenotype, including some that are resistant to conventional chemotherapy due to high BCL-2 expression. We show here that 1) stable BCL-2 overexpression alone can induce a primed for death state and 2) that an ABT-737-induced loss of functional cytochrome c from the electron transport chain causes a reduction in maximal respiration that is readily detectable by microplate-based respirometry. Stable BCL-2 overexpression sensitized non-tumorigenic MCF10A mammary epithelial cells to ABT-737-induced caspase-dependent apoptosis. Mitochondria within permeabilized BCL-2 overexpressing cells were selectively vulnerable to ABT-737-induced cytochrome c release compared to those from control-transfected cells, consistent with a primed state. ABT-737 treatment caused a dose-dependent impairment of maximal O2 consumption in MCF10A BCL-2 overexpressing cells but not in control-transfected cells or in immortalized mouse embryonic fibroblasts lacking both BAX and BAK. This impairment was rescued by delivering exogenous cytochrome c to mitochondria via saponin-mediated plasma membrane permeabilization. An ABT-737-induced reduction in maximal O2 consumption was also detectable in SP53, JeKo-1, and WEHI-231 B-cell lymphoma cell lines, with sensitivity correlating with BCL-2:MCL-1 ratio and with susceptibility (SP53 and JeKo-1) or resistance (WEHI-231) to ABT-737-induced apoptosis. Multiplexing respirometry assays to ELISA-based determination of cytochrome c redistribution confirmed that respiratory inhibition was associated with cytochrome c release. In summary, cell-based respiration assays were able to rapidly identify a primed for death state in cells with either artificially overexpressed or high endogenous BCL-2. Rapid detection of a primed for death state in individual cancers

  6. Validation of an atomic absorption rubidium ion efflux assay for KCNQ/M-channels using the ion Channel Reader 8000.

    PubMed

    Wang, Kewei; McIlvain, Beal; Tseng, Eugene; Kowal, Dianne; Jow, Flora; Shen, Ru; Zhang, Howard; Shan, Qin Jennifer; He, Lan; Chen, Diana; Lu, Qiang; Dunlop, John

    2004-10-01

    M-channels (M-current), encoded by KCNQ2/3 K(+) channel genes, have emerged as novel drug targets for a number of neurological disorders. The lack of direct high throughput assays combined with the low throughput of conventional electrophysiology (EP) has impeded rapid screening and evaluation of K(+)-channel modulators. Development of a sensitive and efficient assay for the direct measurement of M-current activity is critical for identifying novel M-channel modulators and subsequent investigation of their therapeutic potential. Using a stable CHO cell line expressing rat KCNQ2/3 K(+) channels confirmed by EP, we have developed and validated a nonradioactive rubidium (Rb(+)) efflux assay in a 96-well plate format. The Rb(+) efflux assay directly measures the activity of functional channels by atomic absorption spectroscopy using the automated Ion Channel Reader (ICR) 8000. The stimulated Rb(+) efflux from KCNQ2/3-expressing cells was blocked by the channel blockers XE991 and linopirdine with IC(50) values of 0.15 microM and 1.3 microM, respectively. Twelve compounds identified as KCNQ2/3 openers were further assessed in this assay, and their EC(50) values were compared with those obtained with EP. A higher positive correlation coefficient between these two assays (r = 0.60) was observed than that between FlexStation membrane potential and EP assays (r = 0.23). To simplify the assay and increase the throughput, we demonstrate that EC(50) values obtained by measuring Rb(+) levels in the supernatant are as robust and consistent as those obtained from the ratio of Rb(+) in supernatant/lysate. By measuring the supernatant only, the throughput of ICR8000 in an eight-point titration is estimated to be 40 compounds per day, which is suitable for a secondary confirmation assay.

  7. HIGHLY SENSITIVE ASSAY FOR ANTICHOLINESTERASE COMPOUNDS USING 96 WELL PLATE FORMAT

    EPA Science Inventory

    One of the approaches for reducing uncertainties in the assessment of human exposure is to better characterize concentrations of hazardous compounds that may be present in our immediate environment. A significant limitation to this approach, however, is that sampling and labora...

  8. A method for high throughput determination of viable bacteria cell counts in 96-well plates

    PubMed Central

    2012-01-01

    Background There are several methods for quantitating bacterial cells, each with advantages and disadvantages. The most common method is bacterial plating, which has the advantage of allowing live cell assessment through colony forming unit (CFU) counts but is not well suited for high throughput screening (HTS). On the other hand, spectrophotometry is adaptable to HTS applications but does not differentiate between dead and living bacteria and has low sensitivity. Results Here, we report a bacterial cell counting method termed Start Growth Time (SGT) that allows rapid and serial quantification of the absolute or relative number of live cells in a bacterial culture in a high throughput manner. We combined the methodology of quantitative polymerase chain reaction (qPCR) calculations with a previously described qualitative method of bacterial growth determination to develop an improved quantitative method. We show that SGT detects only live bacteria and is sensitive enough to differentiate between 40 and 400 cells/mL. SGT is based on the re-growth time required by a growing cell culture to reach a threshold, and the notion that this time is proportional to the number of cells in the initial inoculum. We show several applications of SGT, including assessment of antibiotic effects on cell viability and determination of an antibiotic tolerant subpopulation fraction within a cell population. SGT results do not differ significantly from results obtained by CFU counts. Conclusion SGT is a relatively quick, highly sensitive, reproducible and non-laborious method that can be used in HTS settings to longitudinally assess live cells in bacterial cell cultures. PMID:23148795

  9. Development of a 96-well plate iodine binding assay for amylose content determination

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cereal starch amylose/amylopectin (AM/AP) ratios are critical in functional properties for food and industrial applications. Conventional methods for the determination of AM/AP of cereal starches are very time consuming and labor intensive making it very difficult to screen large sample sets. Stud...

  10. Determining cereal starch amylose content using a dual wavelength iodine binding 96 well plate assay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cereal starch amylose/amylopectin (AM/AP) ratios are critical in functional properties for food and industrial applications. Conventional determination of AM/AP of cereal starches are very time consuming and labor intensive making it very difficult to screen large sample sets. Studying these large...

  11. Folate content in fresh-cut vegetable packed products by 96-well microtiter plate microbiological assay.

    PubMed

    Fajardo, Violeta; Alonso-Aperte, Elena; Varela-Moreiras, Gregorio

    2015-02-15

    Ready-to-eat foods have nowadays become a significant portion of the diet. Accordingly, nutritional composition of these food categories should be well-known, in particular its folate content. However, there is a broad lack of folate data in food composition tables and databases. A total of 21 fresh-cut vegetable and fruit packed products were analysed for total folate (TF) content using a validated method that relies on the folate-dependent growth of chloramphenicol-resistant Lactobacillus casei subspecies rhamnosus (NCIMB 10463). Mean TF content ranged from 10.0 to 140.9μg/100g for the different matrices on a fresh weight basis. Higher TF quantity, 140.9-70.1μg/100g, was found in spinach, rocket, watercress, chard and broccoli. Significant differences were observed between available data for fresh vegetables and fruits from food composition tables or databases and the analysed results for fresh-cut packed products. Supplied data support the potential of folate-rich fresh-cut ready-to-eat vegetables to increase folate intake significantly.

  12. Caring for Stories: Increasing Reader Engagement in School Libraries

    ERIC Educational Resources Information Center

    Sturm, Brian W.

    2012-01-01

    Librarians, teachers, authors, video game designers, corporate executives, in fact all providers of information struggle with the dilemma of how to get consumers of information engaged with their message. School librarians know that engaged students learn more and retain the information longer; authors and game designers want their readers and…

  13. Crime and Justice in America. A Courses by Newspaper Reader.

    ERIC Educational Resources Information Center

    Skolnick, Jerome H., Ed.; And Others

    This reader is one of several supplementary materials for a 15-week newspaper course about crime and justice, mainly in the United States. Six units contain 67 readings from primary sources such as journal articles, monographs, personal interviews, published letters, and government reports. The readings present personal experiences, research…

  14. 21 CFR 866.2850 - Automated zone reader.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Automated zone reader. 866.2850 Section 866.2850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2850 Automated zone...

  15. 21 CFR 866.2850 - Automated zone reader.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Automated zone reader. 866.2850 Section 866.2850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2850 Automated zone...

  16. 21 CFR 866.2850 - Automated zone reader.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Automated zone reader. 866.2850 Section 866.2850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2850 Automated zone...

  17. 21 CFR 866.2850 - Automated zone reader.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Automated zone reader. 866.2850 Section 866.2850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2850 Automated zone...

  18. 21 CFR 866.2850 - Automated zone reader.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated zone reader. 866.2850 Section 866.2850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2850 Automated zone...

  19. Mnemonic Strategy Instruction for Beginning Readers with Cognitive Impairments

    ERIC Educational Resources Information Center

    Burton, Rivka

    2013-01-01

    Many students with cognitive impairments are not afforded the opportunity to develop their potential as readers. A review of the literature reveals that few researchers have evaluated the effects of phonics instruction on the reading skills of students with cognitive impairments. The purpose of this study was to test the effectiveness of a…

  20. Learning about Mason: A Collaborative Lesson with a Struggling Reader

    ERIC Educational Resources Information Center

    Compton-Lilly, Catherine

    2010-01-01

    In this "Struggling Readers" department, the author describes a collaborative lesson--a professional development strategy adapted from Reading Recovery. Collaborative lessons involve a small group of educators who collaborate to help a student who is struggling with reading. While collaborative lessons were initially designed by Reading Recovery…