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Sample records for 96-well microplate reader

  1. Location dependent biases in automatic 96-well microplate readers.

    PubMed

    Harrison, R O; Hammock, B D

    1988-01-01

    Procedures performed in 96-well microplates and quantitated by automatic readers assume instruments to be precise, accurate, and free of well location dependent bias. Instrument specifications generally focus on precision and accuracy without specifically addressing biases which are dependent on well location. These biases appear to be meniscus dependent and can be demonstrated in varying degrees in automatic readers of many designs by using a reverse plate wet test, which compares repeated readings of a dye loaded plate in normal and reversed positions. This test analyzes differences between readings and is, therefore, independent of pipetting error or other experimental variables such as protein binding or immunoassay variability. Different plates increased or decreased the magnitude of observed errors but did not themselves cause the errors measured by the reverse plate wet test. Error patterns were consistent for each reader and varied widely among the 16 instruments tested. Only 4 of 16 instruments passed an existing manufacturer's specification for precision, and only one of the 16 readers tested passed a similar specification for accuracy. The severest location dependent bias was found in an instrument which exhibited excellent repeatability and consistently passed its built-in diagnostic tests. One reader with significant bias was returned to the manufacturer for routine service and calibration, but it was not demonstrably improved. The reverse plate wet test is an extremely useful diagnostic tool for quality control at all stages of instrument manufacture and use. PMID:3069835

  2. Single-step method for β-galactosidase assays in Escherichia coli using a 96-well microplate reader.

    PubMed

    Schaefer, Jorrit; Jovanovic, Goran; Kotta-Loizou, Ioly; Buck, Martin

    2016-06-15

    Historically, the lacZ gene is one of the most universally used reporters of gene expression in molecular biology. Its activity can be quantified using an artificial substrate, o-nitrophenyl-ß-d-galactopyranoside (ONPG). However, the traditional method for measuring LacZ activity (first described by J. H. Miller in 1972) can be challenging for a large number of samples, is prone to variability, and involves hazardous compounds for lysis (e.g., chloroform, toluene). Here we describe a single-step assay using a 96-well microplate reader with a proven alternative cell permeabilization method. This modified protocol reduces handling time by 90%. PMID:27036618

  3. Single-step method for β-galactosidase assays in Escherichia coli using a 96-well microplate reader

    PubMed Central

    Schaefer, Jorrit; Jovanovic, Goran; Kotta-Loizou, Ioly; Buck, Martin

    2016-01-01

    Historically, the lacZ gene is one of the most universally used reporters of gene expression in molecular biology. Its activity can be quantified using an artificial substrate, o-nitrophenyl-ß-d-galactopyranoside (ONPG). However, the traditional method for measuring LacZ activity (first described by J. H. Miller in 1972) can be challenging for a large number of samples, is prone to variability, and involves hazardous compounds for lysis (e.g., chloroform, toluene). Here we describe a single-step assay using a 96-well microplate reader with a proven alternative cell permeabilization method. This modified protocol reduces handling time by 90%. PMID:27036618

  4. Development of a fluorimetric multispecies 96-well micro-plate growth test for screening metal toxicity to phytoplankton

    SciTech Connect

    Peterson, H.G.; Ruecker, N.J.; Cantin, I.A.; Nyholm, N.; Dal-Jensen, S.

    1995-12-31

    The rapid and cost-effective screening of industrial waste is an ideal approach to regulations that offer true protection of aquatic habitats. For these tests to be ecologically important protection of large groups of organisms is also essential. This can best be done by testing batteries of species. Photosynthetic organisms compose 99.9% of habitats as well as providing food for higher trophic levels. A test was developed that can accommodate the testing of most phytoplanktonic species irrespective of morphology (unicellular, multicellular, colonial, filamentous). Forty eight to 72 h growth tests were carried out with green algae, diatoms, and cyanobacteria. The algae were incubated with different levels of toxicants in 96-well microplates which were read in a 96-well fluorometric plate reader. Phytoplankton emitting low levels of fluorescence can be incubated with DCMU, which can increase the fluorescent signal 2 to 4 times. The data from the plate reader is transferred to a computer spreadsheet and inhibition levels are automatically calculated. Eleven metal mining wastes from across Canada were tested against this method using the following phytoplanktonic species: Selenastrum, Nannochloris (green algae), Nitzschia (diatom), Microcystis, and Pseudoanabaena (cyanobacteria). These wastes were also screened against Microtox. All wastes were highly toxic to the tested phytoplankton, but only 4 were toxic to Microtox{trademark}.

  5. Establishment and validation of a method for multi-dose irradiation of cells in 96-well microplates

    SciTech Connect

    Abatzoglou, Ioannis; Zois, Christos E.; Pouliliou, Stamatia

    2013-02-15

    Highlights: ► We established a method for multi-dose irradiation of cell cultures within a 96-well plate. ► Equations to adjust to preferable dose levels are produced and provided. ► Up to eight different dose levels can be tested in one microplate. ► This method results in fast and reliable estimation of radiation dose–response curves. -- Abstract: Microplates are useful tools in chemistry, biotechnology and molecular biology. In radiobiology research, these can be also applied to assess the effect of a certain radiation dose delivered to the whole microplate, to test radio-sensitivity, radio-sensitization or radio-protection. Whether different radiation doses can be accurately applied to a single 96-well plate to further facilitate and accelerated research by one hand and spare funds on the other, is a question dealt in the current paper. Following repeated ion-chamber, TLD and radiotherapy planning dosimetry we established a method for multi-dose irradiation of cell cultures within a 96-well plate, which allows an accurate delivery of desired doses in sequential columns of the microplate. Up to eight different dose levels can be tested in one microplate. This method results in fast and reliable estimation of radiation dose–response curves.

  6. Protein farnesyltransferase: measurement of enzymatic activity in 96-well format using TopCount microplate scintillation counting technology.

    PubMed

    Harwood, H J

    1995-04-10

    Protein farnesyltransferase (PFT) catalyzes the transfer of the farnesyl group of farnesyl pyrophosphate (FPP) to proteins ending with a carboxy-terminal CAAX motif, forming a thioether linkage to the cysteine residue of the protein. A method is described herein for measurement of PFT activity in 96-well format using TopCount microplate scintillation counting technology. This method has the advantages of requiring only a single transfer from reaction vessels or wells of a 96-well reaction plate to the filtration wells of a 96-well Packard UniFilter GF/B filtration plate following acid precipitation and of allowing liquid scintillation counting to be conducted directly in the filtration plate without the need for either removal of the filter from the plate or transfer of the filter to liquid scintillation vials prior to radioactivity assessment. Using rat brain cytosol as the source of PFT, H-ras as the source of farnesyl acceptor protein, and [1-3H]FPP as the farnesyl donor, the incorporation of radiolabeled farnesyl residues into H-ras was found to be a linear function of both the time of incubation at 37 degrees C (up to 75 min) and the concentration of rat brain cytosolic protein present during incubation (up to 40 micrograms protein), and to be dependent on the concentration of H-ras (Km = 1.1 microM) and FPP (Km = 0.6 microM) present in the incubation reaction. In the presence of 4 microM H-ras, 0.5 microM FPP, 4 mM MgCl2, and 20 microM ZnCl2, the specific activity of rat brain cytosolic PFT measured using this methodology was 0.253 +/- 0.036 (SD; n = 30) pmol H-ras farnesylated per minute of incubation at 37 degrees C per milligram cytosolic protein. The signal-to-noise ratio for H-ras farnesylation using this methodology averaged 25 relative to incubation in the absence of H-ras (background farnesylation of cytosolic proteins) and 50 relative to incubation in the absence of both H-ras and rat brain cytosol (background filter associated radioactivity

  7. 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.

  8. Development of a microplate reader compatible microfluidic chip for ELISA.

    PubMed

    Hou, Fenghua; Zhang, Qin; Yang, Jianping; Li, Xinchun; Yang, Xiujuan; Wang, Shuping; Cheng, Zhiyi

    2012-08-01

    We report a novel microfluidic device use for sandwich enzyme-linked immunoassay assay (ELISA). The related procedures including the introduction of reagents, dilution and distribution of samples, as well as immobilization of enzyme can be readily carried out on a poly (dimethylsiloxane) (PDMS) chip. Particularly, this microfluidic chip comprising of two distinct parallel units, and has an identical dimension as a conventional microtiter plate, which offers access to the directly quantitative detection by the microplate reader. Gradient-concentration reacting solutions at six different concentrations level generated by the microfluidic channel network are simultaneously transported to 24 reaction chambers to form enzymatic products. Alkaline phosphatase (ALP), 4-methylumbelliferyl phosphate (4-MUP) and KH(2)PO(4) are used as enzyme-substrate-inhibitor model, to demonstrate the utility of the developed microchip-based enzyme inhibitor assay. Various conditions such as the surface treatment of chip channels, fluids velocities, substrate concentration, and buffer pH are investigated. The present microfluidic device for ELISA holds several advantages, for instance frugal usage of samples and reagents, less of operating time, favorably integrated configuration, ease of manipulation, and could be explored to a variety of high throughput drug screening. PMID:22526682

  9. 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. PMID:27019031

  10. Development of Singlet Oxygen Absorption Capacity (SOAC) Assay Method Using a Microplate Reader.

    PubMed

    Takahashi, Shingo; Iwasaki-Kino, Yuko; Aizawa, Koichi; Terao, Junji; Mukai, Kazuo

    2016-01-01

    Recently, a new assay method that can quantify the singlet oxygen absorption capacity (SOAC) of natural antioxidants and food extracts was developed. The SOAC values were measured in ethanol-chloroform-D2O (50 + 50 + 1, v/v/v) solution at 35°C using a UV-Vis spectrophotometer equipped with a six-channel cell positioner and an electron-temperature control unit. In the present study, measurement of the SOAC values was performed for eight representative carotenoids and three vegetable extracts (tomato, carrot, and red paprika) using a versatile instrument, the microplate reader. A 24-well glass microplate was used for measurements because a plastic microplate, commonly used in the laboratory, dissolves in the ethanol-chloroform-D2O solution. The SOAC values of eight carotenoids and three vegetable extracts measured using a microplate reader were in good agreement with the corresponding values measured using a UV-Vis spectrophotometer, suggesting that the microplate reader is an applicable instrument for the measurement of reliable SOAC values for general antioxidants and food extracts in solution. PMID:26822807

  11. 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. PMID:26159546

  12. 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.

  13. 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.

  14. 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

  15. 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.

  16. Mismatch oxidation assay: detection of DNA mutations using a standard UV/Vis microplate reader.

    PubMed

    Tabone, Tania; Sallmann, Georgina; Cotton, Richard G H

    2009-01-01

    Simple, low-cost mutation detection assays that are suitable for low-throughput analysis are essential for diagnostic applications where the causative mutation may be different in every family. The mismatch oxidation assay is a simple optical absorbance assay to detect nucleotide substitutions, insertions, and deletions in heteroduplex DNA. The method relies on detecting the oxidative modification products of mismatched thymine and cytosine bases by potassium permanganate as it is reduced to manganese dioxide. This approach, unlike other methods commonly used to detect sequence variants, does not require costly labeled probes or primers, toxic chemicals, or a time-consuming electrophoretic separation step. The oxidation rate, and hence the presence of a sequence variant, is detected by measuring the formation of the potassium permanganate reduction product (hypomanganate diester), which absorbs at the 420-nm visible wavelength, using a standard UV/vis microplate reader. PMID:19768598

  17. Upgrading a microplate reader for photobiology and all-optical experiments.

    PubMed

    Richter, Florian; Scheib, Ulrike S; Mehlhorn, Jennifer; Schubert, Roman; Wietek, Jonas; Gernetzki, Oliver; Hegemann, Peter; Mathes, Tilo; Möglich, Andreas

    2015-02-01

    Automation can vastly reduce the cost of experimental labor and thus facilitate high experimental throughput, but little off-the-shelf hardware for the automation of illumination experiments is commercially available. Here, we use inexpensive open-source electronics to add programmable illumination capabilities to a multimode microplate reader. We deploy this setup to characterize light-triggered phenomena in three different sensory photoreceptors. First, we study the photoactivation of Arabidopsis thaliana phytochrome B by light of different wavelengths. Second, we investigate the dark-state recovery kinetics of the Synechocystis sp. blue-light sensor Slr1694 at multiple temperatures and imidazole concentrations; while the kinetics of the W91F mutant of Slr1694 are strongly accelerated by imidazole, the wild-type protein is hardly affected. Third, we determine the light response of the Beggiatoa sp. photoactivatable adenylate cyclase bPAC in Chinese hamster ovary cells. bPAC is activated by blue light in dose-dependent manner with a half-maximal intensity of 0.58 mW cm(-2); intracellular cAMP spikes generated upon bPAC activation decay with a half time of about 5 minutes after light switch-off. Taken together, we present a setup which is easily assembled and which thus offers a facile approach to conducting illumination experiments at high throughput, reproducibility and fidelity. PMID:25373866

  18. Measuring intracellular calcium dynamics of HeLa cells exposed to nitric oxide by microplate fluorescence reader

    NASA Astrophysics Data System (ADS)

    Huang, Yimei; Chen, Jiangxu; Yang, Hongqin; Zheng, Liqin; Wang, Yuhua; Li, Hui; Xie, Shusen

    2012-12-01

    Nitric oxide (NO) has been reported to have the ability to promote or inhibit the proliferation and metastasis of cancer cells. It appears to have an effect on inducing calcium transient, which participates in essential cellular signaling in the physiological and pathological processes. Our work was intended to study the effects of exogenous NO on intracellular calcium dynamics of HeLa cells with Fluo-3, a calcium fluorescent indicator by microplate fluorescence reader. The results showed that after NO donor was injected into the wells, intracellular Ca2+ fluorescence intensity increased significantly compared with that of control group. Furthermore, the calcium transient activated by NO was mainly due to the calcium release from intracellular calcium stores. These would be helpful to further recognize the role of NO involved in cancer cell proliferation and metastasis.

  19. Measuring platelet aggregation with microplate reader. A new technical approach to platelet aggregation studies.

    PubMed

    Fratantoni, J C; Poindexter, B J

    1990-11-01

    Platelet aggregation measurements were done with the use of a commercially available microtiter plate reader with specific modification of the mode of agitation of the samples. Satisfactory aggregation curves were obtained with use of an external horizontal agitator, with an amplitude of 1.3 mm and minimum frequency of 1,360 cycles/minute. With the use of the 96 available wells in the microtiter plates, all test and control platelet samples, with replicates, were observed simultaneously and the output data obtained within 10-15 minutes. The technique was validated by demonstrating the similarity of dose-response curves, obtained with a standard aggregometer and with the microtiter technique, of platelets stimulated by adenosine diphosphate, thrombin, and arachidonic acid. PMID:2239825

  20. Toward Microbioreactor Arrays: A Slow-Responding Oxygen Sensor for Monitoring of Microbial Cultures in Standard 96-Well Plates.

    PubMed

    Glauche, Florian; John, Gernot T; Arain, Sarina; Knepper, Andreas; Neubauer, Antje; Goelling, Detlef; Lang, Christine; Violet, Norman; King, Rudibert; Neubauer, Peter

    2015-08-01

    In this study, a slow-responding chemo-optical sensor for dissolved oxygen (DO) integrated into a 96-well plate was developed. The slow response time ensures that the measured oxygen value does not change much during plate transport to the microplate reader. The sensor therefore permits at-line DO measurement of microbial cultures. Moreover, it eliminates the necessity of individual optical measurement systems for each culture plate, as many plates can be measured successively. Combined with the 96-well format, this increases the experimental throughput enormously. The novel sensor plate (Slow OxoPlate) consists of fluorophores suspended in a polymer matrix that were placed into u-bottom 96-well plates. Response time was measured using sodium sulfite, and a t90 value of 9.7 min was recorded. For application, DO values were then measured in Escherichia coli and Saccharomyces cerevisiae cultures grown under fed-batch-like conditions. Depending on the DO sensor's response time, different information on the oxygenation state of the culture plate was obtained: a fast sensor variant detects disturbance through sampling, whereas the slow sensor indicates oxygen limitation during incubation. A combination of the commercially available OxoPlate and the Slow OxoPlate enables operators of screening facilities to validate their cultivation procedures with regard to oxygen availability. PMID:25720599

  1. Quantification of cells cultured on 96-well plates.

    PubMed

    Kueng, W; Silber, E; Eppenberger, U

    1989-10-01

    The method for cell number measurement in monolayer cultures by crystal violet staining published recently by Gillies et al. (R. G. Gillies, N. Didier, M. Denton (1986) Anal. Biochem. 159, 109-113) was modified and significantly improved. The procedure was adapted for use in 96-well plates since the method is inherently very sensitive. Modifications allowed fast and complete solubilization of dye adsorbed by cell nuclei during staining. Since light absorption of the unstained or destained cell layers is negligible, cell number measurements can be performed in the respective wells. Due to these features, multiple assays may be carried out rapidly using standard 96-well plate readers. In addition, it is shown that the sensitivity of the assay can be varied and easily controlled by choosing the appropriate pH during the staining procedure. This increases the flexibility of the method making it useful for determining cell density of a wide range of different cell types. PMID:2604040

  2. 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.

  3. Lactobacillus casei microbiological assay of folic acid derivatives in 96-well microtiter plates.

    PubMed

    Horne, D W; Patterson, D

    1988-11-01

    Microbiological assay is still widely used for estimating folic acid derivatives in serum and other biological samples. We describe here a modification of this procedure involving use of 96-well microtiter plates. This procedure, used with modern, computer-interfaced microtiter-plate readers and data-reduction software, greatly shortens the time and minimizes reagent costs for this assay. Under the conditions of our assay procedures, all folic acid derivatives tested gave equal growth response for Lactobacillus casei. Results for assays of rat liver extracts showed excellent agreement between the standard bioassay and the 96-well procedure. PMID:3141087

  4. Automation of cell-based drug absorption assays in 96-well format using permeable support systems.

    PubMed

    Larson, Brad; Banks, Peter; Sherman, Hilary; Rothenberg, Mark

    2012-06-01

    Cell-based drug absorption assays, such as Caco-2 and MDCK-MDR1, are an essential component of lead compound ADME/Tox testing. The permeability and transport data they provide can determine whether a compound continues in the drug discovery process. Current methods typically incorporate 24-well microplates and are performed manually. Yet the need to generate absorption data earlier in the drug discovery process, on an increasing number of compounds, is driving the use of higher density plates. A simple, more efficient process that incorporates 96-well permeable supports and proper instrumentation in an automated process provides more reproducible data compared to manual methods. Here we demonstrate the ability to perform drug permeability and transport assays using Caco-2 or MDCKII-MDR1 cells. The assay procedure was automated in a 96-well format, including cell seeding, media and buffer exchanges, compound dispense, and sample removal using simple robotic instrumentation. Cell monolayer integrity was confirmed via transepithelial electrical resistance and Lucifer yellow measurements. Proper cell function was validated by analyzing apical-to-basolateral and basolateral-to-apical movement of rhodamine 123, a known P-glycoprotein substrate. Apparent permeability and efflux data demonstrate how the automated procedure provides a less variable method than manual processing, and delivers a more accurate assessment of a compound's absorption characteristics. PMID:22357561

  5. Detection of 100% of mutations in 124 individuals using a standard UV/Vis microplate reader: a novel concept for mutation scanning

    PubMed Central

    Tabone, Tania; Sallmann, Georgina; Webb, Elizabeth; Cotton, Richard G. H.

    2006-01-01

    We report the development of a simple and inexpensive assay for the detection of DNA polymorphisms and mutations that is based on the modification of mismatched bases by potassium permanganate. Unlike the chemical cleavage of mismatch assay, which also exploits the reactivity of potassium permanganate to detect genomic variants, the assay we describe here does not require a cleavage manipulation and therefore does not require expensive or toxic chemicals or a separation step, as mismatches are detected using direct optical methods in a microplate format. Studies with individual deoxynucleotides demonstrated that the reactivity with potassium permanganate resulted in a specific colour change. Furthermore, studies with synthetic oligonucleotide heteroduplexes demonstrated that this colour change phenomenon could be applied to detect mismatched bases spectrophotometrically. A collection of plasmids carrying single point mutations in the mouse β-globin promoter region was used as a model system to develop a functional mutation detection assay. Finally, the assay was validated as 100% effective in detecting mismatches in a blinded manner using DNA from patients previously screened for mutations using established techniques, such as sequencing, SSCP and denaturing high-performance liquid chromatography (DHPLC) analysis in DNA fragments up to 300 bp in length. PMID:16554551

  6. Laboratory evaluation of a SpectraMax microplate reader and test strips for field measurement of creatinine in spot urine samples in the event of a radiological accident.

    PubMed

    Daka, Joseph N; Moodie, Gerry; Li, Chunsheng; Wilkins, Ruth; Kramer, Gary H

    2011-08-01

    The fear that terrorists might use radiological or nuclear (RN) devices to attack others is a new but growing phenomenon, arising mainly from the events of 11 September 2001. Research on rapid analytical methods that can allow analyses of large numbers of people who may become internally contaminated with radionuclides due to a RN accident is still limited. To contribute to this bioassay capacity for emergency response, the Radiation Protection Bureau of Health Canada has identified and evaluated two new portable SpectraMax plate readers (model 250 and Plus 384) and one brand of dry reagent strips for rapid measurement of creatinine in spot urine samples. Concentrations of creatinine in spot urine samples provide a means of adjusting or normalizing urine collections to 24 h, upon which accurate internal dose assessments due to the radionuclides can be made. Preliminary test results of the devices showed the two SpectraMax plate readers and the TECO dry creatinine reagent strips were portable, rapid and reliable for urinary creatinine measurements in spot samples, suggesting they can be used in rapid dose screening of people. PMID:21709503

  7. 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…

  8. A microassay for measuring glycogen in 96-well-cultured cells.

    PubMed

    Gómez-Lechón, M J; Ponsoda, X; Castell, J V

    1996-05-01

    This study describes a rapid, sensitive, and automated spectrophotometric enzymatic microassay that measures the intracellular glycogen of primary cultured hepatocytes and other cultured cells in 96-well plates and can be adapted for other samples that are transferred to these plates. The procedure involves in situ disruption of cells, followed by hydrolysis of glycogen into glucosyl units by fungal glucoamylase (exo-1.4-alpha-glucosidase, EC 3.2.1.3), and glucose determination with the glucose oxidase colorimetric method. The color intensity can be measured in conventional ELISA readers, and the data can be fed to an on-line computer for rapid processing. The advantages of this method are its simplicity and automation, the reduction in sample handling, and the small number of cells required compared to other conventional methods. PMID:8660508

  9. Optimization of phagocyte chemiluminescence measurements using microplates and vials.

    PubMed

    Hengster, P; Kunc, M; Linke, R; Eberl, T; Steurer, W; Ofner, D; Berthold, F; Margreiter, R

    1999-01-01

    In order to cope with large amounts of samples for chemiluminescence (CL), vials were replaced with microplates. Although various types of plates have been commercially available for quite some time and the free-plate mode is advocated by the producer of the counter, little is known about their impact on the outcome of CL measurements. We tested two different 24-well microplates and six different 96-well microplates in two different luminometers, and results were compared with those achieved with vials. Before these comparative tests, we attempted to optimize measurement conditions. CL sensitivity was highest with luminol concentrations of 0.8-3.3 micromol/L, PMA concentrations of 0.06-80 micromol/L, a pH value of 10 and a temperature of 20 degrees C. An indirect correlation was found between fluid volume and yield in counts: the lower the volume, the higher the counts. With regard to sensitivity and cross-talk, the 96-well Isoplatetrade mark was superior to all other plates tested. While all white plates tested gave acceptable results, usage of the black 96-well plates resulted in an extremely low sensitivity. Plates designed for cell culturing gave even lower counts and a cross-talk of up to 31%. All attempts to reduce cross-talk and improve sensitivity, such as aluminium foil or grids, irrespective of the position of the photomultiplier, did not give results comparable to the original 96-well isoplate. Our results suggest that, with the exception of black 96-well microplates and cell culture plates, all other plates tested have a sufficient sensitivity when compared to vials and acceptable cross-talk, the 96-well Isoplatetrade mark being the best. Both types of luminometers used gave reproducible results, Wallac having a somewhat higher sensitivity, Canberra Packard somewhat less cross-talk. PMID:10398566

  10. Rapid acid digestion and simple microplate method for milk iodine determination.

    PubMed

    Hedayati, Mehdi; Ordookhani, Arash; Daneshpour, Maryam Sadat; Azizi, Fereidoun

    2007-01-01

    Iodine deficiency leads to deficiency of thyroid hormones, which causes mental retardation in infant. Laboratory confirmation is important in its diagnosis. The major problems associated with the existing methods for iodine determination in milk samples are: 1) nonsafe alkaline solution; 2) harsh thermal condition; and 3) extra time required to complete various steps. In this study, a simple and rapid colorimetric method was investigated, which used acid digestion in combination with a rapid microplate reading format method to determine the total iodine content in milk. Sample digestion was done on 50 microL milk in metavanadate/perchloric, at 230 degrees C for 10 min. After digestion, iodine determination was based on the Sandell-Kolthoff reaction. The reaction results were read in 96-well microplates by an enzyme-linked immunosorbent assay (ELISA) reader. The determination range of the assay was between 2 and 40 microg/dL. The within-run coefficient of variation percent in three levels (3, 12, and 36 microg/dL) ranged from 6.7 to 9.3 and between-run coefficients of variation ranged from 8.6 to 12.3%. The results obtained (n=70) by the optimized method have good correlation with the results of alkaline incineration as a reference method (n=70; r2=0.907; y=0.952x+1.77). Recovery tests for accuracy assessment in six levels from 6.2 to 34.2 microg/dL) were between 91.3 and 113%. This method has enabled us to achieve 0.12 microg/dL sensitivity. The results of this study show that a quick acid digestion combined with mild thermal and low sample volume with a quick reading of assay results were the main advantages of the acid digestion and microplate reading format. PMID:17847102

  11. The Galapagos Microplate Revealed

    NASA Astrophysics Data System (ADS)

    Smith, D. K.; Schouten, H.; Cann, J. R.; Zhu, W.; Montesi, L. G.; Mitchell, G. A.

    2009-12-01

    We report a new bathymetry survey of the Galapagos microplate (GMP), which separates the Pacific, Nazca, and Cocos plates at the Galapagos Triple Junction. Prior to the formation of the microplate, 1.5-1.0 Ma, there was a succession of transient minor rifts forming triple junctions north and south of the propagating Cocos-Nazca rift (see Schouten et al. abstract). As proposed by Lonsdale (1988) the formation of a large near-axis seamount coincided with the initiation of the GMP and stabilized rifting on its southern boundary, now called Dietz Deep Rift. Lonsdale also proposed that the GMP was rotating clockwise at 6 degrees/my. Schouten et al. (1993) and Klein et al. (2005) applied an edge-driven microplate model to the GMP to understand its kinematics and predicted rotation rates of 30-40 degrees/my and 22 degrees/my, respectively. These interpretations and predictions were based on sparse bathymetry data. In early 2009 (AT 15-41), we mapped the Galapagos microplate in its entirety to understand more fully the conditions that led to the stabilization of the southern triple junction at Dietz Deep Rift and to constrain the rotation rate of the microplate. Our new data show the two highly contrasted sections of Dietz Deep Rift. The northeastern section contains Dietz Deep, a 2 km deep basin, within a fault-dominated rift valley about 20 km wide; subsidiary rifts occur to the south. Sidescan data indicate that extension in this broadly rifted area has been largely amagmatic. The southwestern section of Dietz Deep Rift is dominated by a variety of volcanic constructions in which faulting plays a minor part. The volcanism has resulted in two large seamounts and a number of volcanic ridges running parallel to the fault dominated rift valley. The largest volcanic ridge is steep-sided and straight, and extends to intersect the East Pacific Rise (EPR) at 1 10’N to form the triple junction. Other minor volcanic ridges occur in the SW section of the microplate fanning

  12. 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...

  13. 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. PMID:12790681

  14. 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...

  15. IDENTIFICATION OF ANTIFUNGAL COMPOUNDS IN A BIOLOGICAL CONTROL PRODUCT USING A MICROPLATE INHIBITION BIOASSAY AND MS ANALYSIS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An inhibition assay was developed to quantify the antifungal activity of a solid state fermentation of Bacillus subtilis. Methanol extracts were tested against a spore solution of the fungal pathogen Fusarium oxysporum f. sp. melonis using a 96-well microplate bioassay. HPLC analysis of extracts s...

  16. 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...

  17. 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.

  18. 96-well format-based microfluidic platform for parallel interconnection of multiple multicellular spheroids.

    PubMed

    Kim, Jin-Young; Fluri, David A; Kelm, Jens M; Hierlemann, Andreas; Frey, Olivier

    2015-06-01

    In this article, we present a microfluidic platform, compatible with conventional 96-well formats, that enables facile and parallelized culturing and testing of spherical microtissues in a standard incubator. The platform can accommodate multiple microtissues (up to 66) of different cell types, formed externally by using the hanging-drop method, and enables microtissue interconnection through microfluidic channels for continuous media perfusion or dosage of substances. The platform contains 11 separate channels, and each channel has six tissue compartments. Primary rat liver tissues were cultured over 8 days, and multiple tumor tissues (HCT116) were exposed to various concentrations of 5-fluorouracil for platform characterization. PMID:25524491

  19. Semi-automated microplate monitoring of protein polymerization and aggregation.

    PubMed

    Garcia, Veronica M; Rowlett, Veronica W; Margolin, William; Morano, Kevin A

    2016-09-01

    Static light scattering (SLS) is a commonly used technique for monitoring dynamics of high molecular weight protein complexes such as protein oligomers or aggregates. However, traditional methods are limited to testing a single condition and typically require large amounts of protein and specialized equipment. We show that a standard microplate reader can be used to characterize the molecular dynamics of different types of protein complexes, with the multiple advantages of microscale experimental volumes, semi-automated protocols and highly parallel processing. PMID:27251433

  20. Biofilm Localization in the Vertical Wall of Shaking 96-Well Plates

    PubMed Central

    Gomes, Luciana C.; Moreira, Joana M. R.; Simões, Manuel; Melo, Luís F.; Mergulhão, Filipe J.

    2014-01-01

    Microtiter plates with 96 wells are being increasingly used for biofilm studies due to their high throughput, low cost, easy handling, and easy application of several analytical methods to evaluate different biofilm parameters. These methods provide bulk information about the biofilm formed in each well but lack in detail, namely, regarding the spatial location of the biofilms. This location can be obtained by microscopy observation using optical and electron microscopes, but these techniques have lower throughput and higher cost and are subjected to equipment availability. This work describes a differential crystal violet (CV) staining method that enabled the determination of the spatial location of Escherichia coli biofilms formed in the vertical wall of shaking 96-well plates. It was shown that the biofilms were unevenly distributed on the wall with denser cell accumulation near the air-liquid interface. The results were corroborated by scanning electron microscopy and a correlation was found between biofilm accumulation and the wall shear strain rates determined by computational fluid dynamics. The developed method is quicker and less expensive and has a higher throughput than the existing methods available for spatial location of biofilms in microtiter plates. PMID:24834360

  1. 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. PMID:26818699

  2. 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

  3. Stability test for amorphous materials in humidity controlled 96-well plates by near-infrared spectroscopy.

    PubMed

    Otsuka, Makoto; Tanabe, Hideaki

    2012-03-01

    The purpose of this research is to apply near infrared spectrometry (NIR) with chemoinformetrics to predict the change of crystalline properties of indomethacin (IMC) amorphous under various levels of relative humidity storage conditions. Stability test for amorphous and meta-stable polymorphic forms was performed in humidity controlled the modified 96-well quartz plates containing various kinds of saturated salt solutions (0-100% of relative humidity (RH)) by NIR spectroscopy. Amorphous form was obtained melt product to pour into liquid nitrogen and after then ground. Samples were stored at 25°C in the 6-well plates at various levels of RH. The spectra of the powder samples were measured by the reflectance FT-NIR spectrometer. The second derivative spectra of form α showed specific absorption peaks at 4980, 6036, 7296 and 8616 cm-1 and that of form γ showed those at 5020, 5028, 7344, 7428 and 8436 cm-1. After storage at less than 50% RH, the peak intensities at 5020, 5028, 7344, 7428 and 8436 cm-1 of the amorphous solid increased with increasing of storage time. However, the peak intensity at 4980, 6036 and 7296 cm-1 increased at more than 50% RH Please check and confirm the edit. The results suggested that at lower humidity, the IMC amorphous solid transformed into form γ, but it transformed into form α at more than high humidity. It is possible that crystalline stability of the pharmaceutical preparations could be predicted by using humidity controlled 96-well plates and reflectance NIR-chemoinformetric methods. PMID:21942281

  4. A novel 96-well gel-based assay for determining antifungal activity against filamentous fungi.

    PubMed

    Troskie, Anscha Mari; Vlok, Nicolas Maré; Rautenbach, Marina

    2012-12-01

    In recent years the global rise in antibiotic resistance and environmental consciousness lead to a renewed fervour to find and develop novel antibiotics, including antifungals. However, the influence of the environment on antifungal activity is often disregarded and many in vitro assays may cause the activity of certain antifungals to be overestimated or underestimated. The general antifungal test assays that are economically accessible to the majority of scientists primarily rely on visual examination or on spectrophotometric analysis. The effect of certain morphogenic antifungals, which may lead to hyperbranching of filamentous fungi, unfortunately renders these methods unreliable. To minimise the difficulties experienced as a result of hyperbranching, we developed a straightforward, economical 96-well gel-based method, independent of spectrophotometric analysis, for highly repeatable determination of antifungal activity. For the calculation of inhibition parameters, this method relies on the visualisation of assay results by digitisation. The antifungal activity results from our novel micro-gel dilution assay are comparable to that of the micro-broth dilution assay used as standard reference test of The Clinical and Laboratory Standard Institute. Furthermore, our economical assay is multifunctional as it permits microscopic analysis of the preserved assay results, as well as rendering highly reliable data. PMID:23089670

  5. Measuring oxidative stress resistance of Caenorhabditis elegans in 96-well microtiter plates.

    PubMed

    Possik, Elite; Pause, Arnim

    2015-01-01

    Oxidative stress, which is the result of an imbalance between production and detoxification of reactive oxygen species, is a major contributor to chronic human disorders, including cardiovascular and neurodegenerative diseases, diabetes, aging, and cancer. Therefore, it is important to study oxidative stress not only in cell systems but also using whole organisms. C. elegans is an attractive model organism to study the genetics of oxidative stress signal transduction pathways, which are highly evolutionarily conserved. Here, we provide a protocol to measure oxidative stress resistance in C. elegans in liquid. Briefly, ROS-inducing reagents such as paraquat (PQ) and H2O2 are dissolved in M9 buffer, and solutions are aliquoted in the wells of a 96 well microtiter plate. Synchronized L4/young adult C. elegans animals are transferred to the wells (5-8 animals/well) and survival is measured every hour until most worms are dead. When performing an oxidative stress resistance assay using a low concentration of stressors in plates, aging might influence the behavior of animals upon oxidative stress, which could lead to an incorrect interpretation of the data. However, in the assay described herein, this problem is unlikely to occur since only L4/young adult animals are being used. Moreover, this protocol is inexpensive and results are obtained in one day, which renders this technique attractive for genetic screens. Overall, this will help to understand oxidative stress signal transduction pathways, which could be translated into better characterization of oxidative stress-associated human disorders. PMID:25993260

  6. A 96-well electrochemical method for the screening of enzymatic activities.

    PubMed

    Abdellaoui, Sofiène; Noiriel, Alexandre; Henkens, Robert; Bonaventura, Celia; Blum, Loïc J; Doumèche, Bastien

    2013-04-01

    The rapid electrochemical screening of enzyme activities in bioelectronics is still a challenging issue. In order to solve this problem, we propose to use a 96-well electrochemical assay. This system is composed of 96 screen-printed electrodes on a printed circuit board adapted from a commercial system (carbon is used as the working electrode and silver chloride as the counter/reference electrode). The associated device allows for the measurements on the 96 electrodes to be performed within a few seconds. In this work, we demonstrate the validity of the screening method with the commercial laccase from the fungus Trametes versicolor. The signal-to-noise ratio (S/N) is found to be the best way to analyze the electrochemical signals. The S/N follows a saturation-like mechanism with a dynamic linear range of two decades ranging from 0.5 to 75 ng of laccase (corresponding to enzymatic activities from 62 × 10(-6) to 9.37 × 10(-3) μmol min(-1)) and a sensitivity of 3027 μg(-1) at +100 mV versus Ag/AgCl. Laccase inhibitors (azide and fluoride anions), pH optima, and interfering molecules could also be identified within a few minutes. PMID:23461701

  7. A rapid direct telomerase assay method using 96-well streptavidin plates.

    PubMed

    Francis, Rawle; Friedman, Simon H

    2002-05-01

    We have developed a high-throughput direct assay methodfor the assay of telomerase activity that improves on previous direct telomerase assays in two ways that allow larger numbers of samples to be conveniently processed: (i) 96-well streptavidin coated plates are used to bind and wash biotinylated primer extension products from the telomerase assay, as opposed to tubes containing streptavidin-coated magnetic beads; and (ii) storage phosphor-imagery is used instead of film autoradiography to detect telomerase products after being washed and released from the streptavidin-derivatized matrix. This method improves on previous direct assay methods using magnetic beads by allowing larger numbers of samples to be conveniently assayed. Also, the total activity of the radiolabeled nucleotides used in this procedure is significantly lower than that used in standard direct telomerase assays, lowering costs and exposure to radioactivity. We have validated the assay by repeating, in triplicate, the IC50 determination of rivanol, our previously identified telomerase inhibitor. PMID:12019789

  8. Establishment of ATP-based luciferase viability assay in 96-well plate for Trypanosoma congolense.

    PubMed

    Suganuma, Keisuke; Allamanda, Puttik; Hakimi, Hassan; Zhou, Mo; Angeles, Jose Ma; Kawazu, Shin-ichiro; Inoue, Noboru

    2014-11-01

    Animal African trypanosomosis (AAT), caused by Trypanosoma congolense, is widespread throughout sub-Saharan Africa. There are significant concerns related to the current drugs available for the treatment of AAT due to their limited effectiveness across species and their adverse effects. Moreover, drug resistant trypanosomes have recently been reported in the field. High throughput screening (HTS) of large chemical compound library collections is a promising approach for identifying novel drug candidates. While HTS for Trypanozoon trypanosomes, T. brucei sspp. and T. evansi is well established, no assays have been developed for T. congolense. In the present study, the authors developed an ATP-based luciferase viability assay for T. congolense in a 96-well plate format. The calculated 50% inhibitory concentration (IC50) values for pentamidine and diminazene were 10-100 times higher in T. congolense than in T. brucei. This result suggests that the transporters for the 2 tested compounds differ between T. congolense and T. brucei. This assay could further be applied to screen novel chemical compounds for the treatment of AAT caused by T. congolense. PMID:25056575

  9. Establishment of ATP-Based Luciferase Viability Assay in 96-Well Plate for Trypanosoma congolense

    PubMed Central

    SUGANUMA, Keisuke; ALLAMANDA, Puttik; HAKIMI, Hassan; ZHOU, Mo; ANGELES, Jose Ma.; KAWAZU, Shin-ichiro; INOUE, Noboru

    2014-01-01

    ABSTRACT Animal African trypanosomosis (AAT), caused by Trypanosoma congolense, is widespread throughout sub-Saharan Africa. There are significant concerns related to the current drugs available for the treatment of AAT due to their limited effectiveness across species and their adverse effects. Moreover, drug resistant trypanosomes have recently been reported in the field. High throughput screening (HTS) of large chemical compound library collections is a promising approach for identifying novel drug candidates. While HTS for Trypanozoon trypanosomes, T. brucei sspp. and T. evansi is well established, no assays have been developed for T. congolense. In the present study, the authors developed an ATP-based luciferase viability assay for T. congolense in a 96-well plate format. The calculated 50% inhibitory concentration (IC50) values for pentamidine and diminazene were 10–100 times higher in T. congolense than in T. brucei. This result suggests that the transporters for the 2 tested compounds differ between T. congolense and T. brucei. This assay could further be applied to screen novel chemical compounds for the treatment of AAT caused by T. congolense. PMID:25056575

  10. 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

    2014-06-01

    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

  11. Human tear analysis with miniaturized multiplex cytokine assay on “wall-less” 96-well plate

    PubMed Central

    Quah, Joanne; Tong, Louis; Kim, Namyong

    2015-01-01

    Purpose Tears are a particularly limited body fluid and commonly used in the diagnosis of patients who have ocular diseases. A popular method for analysis of ocular inflammation in tears uses Luminex® bead multiplex technology to generate valuable multiple cytokine profile outputs with 25–50 µl tear sample volume. We propose a method for measuring tear cytokines with 5 μl tear sample volume and 80% reduced Luminex reagents compared to previous protocols. Methods Using human tears pooled from 1,000 participants, the DA-Bead-based method running at 5–20 µl volume, using manual pipetting, in conjunction with a magnetic Luminex cytokine (four-plex) panel assay in a 96-well format was performed and validated for tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-1β, and IL-6. Results Upon use of the DA-Bead method at the 5 μl volume with cytokine standards, the concentrations of each of the four cytokines were found to be linear over a range of 3.5–4 log pg/ml with an intra-assay coefficient of variation (CV) ≤5%, inter-assay %CV ≤10%, and accuracy within the 70–130% range. Upon use of a 5 µl healthy pooled tear sample, cytokine concentrations were detected with a precision intra-assay %CV ˂ 20% for IL-6, IFN-γ, or TNF-α or 30.37% with IL-1β. The inter-assay %CV with tears was ≤20.84% for all cytokines. Tear volumes run at 5 μl on DA-Bead produced a similar cytokine expression profile at a 1-month interval and were highly correlated with the larger 10 μl–based tear sample volume cytokine profile with R2 = 0.98. Conclusions DA-Bead assay is highly sensitive and reproducible and has a performance profile that is potentially suitable for use in standard clinical scenarios. Considering the use of as little as 5 µl of assay beads and 5 µl sample, this is also likely to reduce the assay cost significantly and ease diagnosis of patients with ocular diseases. PMID:26539027

  12. 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

    2014-08-01

    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. PMID:25022949

  13. MStern Blotting-High Throughput Polyvinylidene Fluoride (PVDF) Membrane-Based Proteomic Sample Preparation for 96-Well Plates.

    PubMed

    Berger, Sebastian T; Ahmed, Saima; Muntel, Jan; Cuevas Polo, Nerea; Bachur, Richard; Kentsis, Alex; Steen, Judith; Steen, Hanno

    2015-10-01

    We describe a 96-well plate compatible membrane-based proteomic sample processing method, which enables the complete processing of 96 samples (or multiples thereof) within a single workday. This method uses a large-pore hydrophobic PVDF membrane that efficiently adsorbs proteins, resulting in fast liquid transfer through the membrane and significantly reduced sample processing times. Low liquid transfer speeds have prevented the useful 96-well plate implementation of FASP as a widely used membrane-based proteomic sample processing method. We validated our approach on whole-cell lysate and urine and cerebrospinal fluid as clinically relevant body fluids. Without compromising peptide and protein identification, our method uses a vacuum manifold and circumvents the need for digest desalting, making our processing method compatible with standard liquid handling robots. In summary, our new method maintains the strengths of FASP and simultaneously overcomes one of the major limitations of FASP without compromising protein identification and quantification. PMID:26223766

  14. 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.

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

    PubMed Central

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

    2009-01-01

    A fluorescence sandwich immunoassay using high affinity antibodies and quantum dot (QD) reporters has been developed for detection of botulinum neurotoxin serotype A (BoNT/A) using a nontoxic recombinant fragment of the holotoxin (BoNT/A-HC-fragment) 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 using AR4 as the capture antibody and QD-coupled RAZ1 as the reporter. Detection to 31 pM with a total incubation time of 3 hours was demonstrated. In a second format, the AR4 capture antibody was coupled to Sepharose beads, and the reactions were carried out in microcentrifuge tubes with an incubation time of 1 hour. The beads were subsequently captured and concentrated in a rotating rod “renewable surface” flow cell equipped with a fiber optic system for fluorescence measurements. In PBS buffer, the BoNT/A-HC-fragment was detected to concentrations as low as 5 pM using the fluidic measurement approach. PMID:19643593

  16. 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.

  17. 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

    2011-01-01

    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

  18. Fast carbohydrate analysis via liquid chromatography coupled with ultra violet and electrospray ionization ion trap detection in 96-well format.

    PubMed

    Rühmann, Broder; Schmid, Jochen; Sieber, Volker

    2014-07-11

    A fast carbohydrate screening platform processible in 96-well format is described. The method is suitable for the determination of various carbohydrates out of complex mixtures as obtained by acidic hydrolysis of carbohydrates polymers. The chromatographic conditions for an efficient separation (12min) and the derivatization process with 1-phenyl-3-methyl-5-pyrazolone (PMP) were optimized for high resolution separation and simultaneous determination of deoxy-, amino-, anhydro-sugars as well as hexoses, pentoses, dimers, uronic acids and degradation products like furfural and hydroxymethylfurfural (HMF). The potential to quantify with UV- and MS-detector in the same range has been demonstrated for 20 different compounds. Finally, the matrix effects of the hydrolysis were positively evaluated. The micro scale hydrolysis and PMP-derivatization without any extraction or drying steps, both in 96-well format, result in a fast and intuitive sample preparation. In combination with a fast liquid chromatography coupled to UV and electrospray ionization ion trap detection (LC-UV-ESI-MS/MS) for the qualification and quantification of various sugars, dimers and degradation products, this method shows great performance in carbohydrate analysis. PMID:24861788

  19. 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.

  20. 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

  1. Microplate Assay for Colletotrichum Spore Production

    PubMed Central

    Slade, S. J.; Harris, R. F.; Smith, C. S.; Andrews, J. H.; Nordheim, E. V.

    1987-01-01

    A simple microplate method was devised to assay spore production by Colletotrichum gloeosporioides by growing the fungus on 1 ml of solid media in the wells of tissue culture plates. Growth and sporulation on microplates were compared at days 4 and 8 with growth and sporulation in 100-ml liquid batch cultures that involved 11 common media. Spore production per unit volume of medium was the same for solid and liquid forms of the media. Qualitative assessment of mycelial growth measured on microplates agreed with that of growth measured in liquid cultures. The microplate assay indicated that V8 juice was the best medium and that an organic content of about 6 mg/ml was optimal for high sporulation and low mycelium production. The assay provides a convenient, rapid, and inexpensive means of screening media for the production of fungal conidia in large numbers, to be used, for example, in biological control programs. PMID:16347310

  2. 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…

  3. 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

  4. 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. PMID:23957476

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

    PubMed

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

    2013-11-15

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

  6. 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

  7. 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

    2015-01-01

    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

  8. High-throughput microplate technique for enzymatic hydrolysis of lignocellulosic biomass.

    PubMed

    Chundawat, Shishir P S; Balan, Venkatesh; Dale, Bruce E

    2008-04-15

    Several factors will influence the viability of a biochemical platform for manufacturing lignocellulosic based fuels and chemicals, for example, genetically engineering energy crops, reducing pre-treatment severity, and minimizing enzyme loading. Past research on biomass conversion has focused largely on acid based pre-treatment technologies that fractionate lignin and hemicellulose from cellulose. However, for alkaline based (e.g., AFEX) and other lower severity pre-treatments it becomes critical to co-hydrolyze cellulose and hemicellulose using an optimized enzyme cocktail. Lignocellulosics are appropriate substrates to assess hydrolytic activity of enzyme mixtures compared to conventional unrealistic substrates (e.g., filter paper, chromogenic, and fluorigenic compounds) for studying synergistic hydrolysis. However, there are few, if any, high-throughput lignocellulosic digestibility analytical platforms for optimizing biomass conversion. The 96-well Biomass Conversion Research Lab (BCRL) microplate method is a high-throughput assay to study digestibility of lignocellulosic biomass as a function of biomass composition, pre-treatment severity, and enzyme composition. The most suitable method for delivering milled biomass to the microplate was through multi-pipetting slurry suspensions. A rapid bio-enzymatic, spectrophotometric assay was used to determine fermentable sugars. The entire procedure was automated using a robotic pipetting workstation. Several parameters that affect hydrolysis in the microplate were studied and optimized (i.e., particle size reduction, slurry solids concentration, glucan loading, mass transfer issues, and time period for hydrolysis). The microplate method was optimized for crystalline cellulose (Avicel) and ammonia fiber expansion (AFEX) pre-treated corn stover. PMID:18306256

  9. BULGARIAN READER.

    ERIC Educational Resources Information Center

    HODGE, CARLETON T.

    DESIGNED TO FOLLOW THE BASIC COURSE IN BULGARIAN, A READER HAS BEEN PREPARED. IT ASSUMES KNOWLEDGE OF THE VOCABULARY (AS PRESENTED IN THE BASIC COURSE ED 003 873 AND ED 003 874) AS WELL AS THE GRAMMATICAL STRUCTURE OF THE LANGUAGE. THE READER PRESENTS AREA MATERIALS AND INTRODUCES THE STUDENT TO THE VOCABULARY NEEDED TO READ NEWSPAPERS AND SIMILAR…

  10. Accelerated Reader.

    ERIC Educational Resources Information Center

    Education Commission of the States, Denver, CO.

    This paper provides an overview of Accelerated Reader, a system of computerized testing and record-keeping that supplements the regular classroom reading program. Accelerated Reader's primary goal is to increase literature-based reading practice. The program offers a computer-aided reading comprehension and management program intended to motivate…

  11. Facile preparation of a photoactivatable surface on a 96-well plate: a versatile and multiplex cell migration assay platform.

    PubMed

    Kamimura, Masao; Scheideler, Olivia; Shimizu, Yoshihisa; Yamamoto, Shota; Yamaguchi, Kazuo; Nakanishi, Jun

    2015-06-01

    Cell migration is an essential cellular activity in various physiological and pathological processes, such as wound healing and cancer metastasis. Therefore, in vitro cell migration assays are important not only for fundamental biological studies but also for evaluating potential drugs that control cell migration activity in medical applications. In this regard, robust control over cell migrating microenvironments is critical for reliable and quantitative analysis as cell migration is highly dependent upon the microenvironments. Here, we developed a facile method for making a commercial glass-bottom 96-well plate photoactivatable for cell adhesion, aiming to develop a versatile and multiplex cell migration assay platform. Cationic poly-d-lysine was adsorbed to the anionic glass surface via electrostatic interactions and, subsequently, functionalized with poly(ethylene glycol) (PEG) bearing a photocleavable reactive group. The initial PEGylated surface is non-cell-adhesive. However, upon near-ultraviolet (UV) irradiation, the photorelease of PEG switches the surface from non-biofouling to cell-adhesive. With this platform, we assayed cell migration in the following procedure: (1) create cell-attaching regions of precise geometries by controlled photoirradiation, (2) seed cells to allow them to attach selectively to the irradiated regions, (3) expose UV light to the remaining PEGylated regions to extend the cell-adhesive area, (4) analyse cell migration using microscopy. Surface modification of the glass surface was characterized by ζ-potential and contact angle measurements. The PEGylated surface showed cell-resistivity and became cell-adhesive upon releasing PEG by near-UV irradiation. The method was applied for parallelly evaluating the effect of model drugs on the migration of epithelial MDCK cells in the multiplexed platform. The dose-response relationship for cytochalasin D treatment on cell migration behavior was successfully evaluated with high

  12. The BUME method: a novel automated chloroform-free 96-well total lipid extraction method for blood plasma[S

    PubMed Central

    Löfgren, Lars; Ståhlman, Marcus; Forsberg, Gun-Britt; Saarinen, Sinikka; Nilsson, Ralf; Hansson, Göran I.

    2012-01-01

    Lipid extraction from biological samples is a critical and often tedious preanalytical step in lipid research. Primarily on the basis of automation criteria, we have developed the BUME method, a novel chloroform-free total lipid extraction method for blood plasma compatible with standard 96-well robots. In only 60 min, 96 samples can be automatically extracted with lipid profiles of commonly analyzed lipid classes almost identically and with absolute recoveries similar or better to what is obtained using the chloroform-based reference method. Lipid recoveries were linear from 10–100 µl plasma for all investigated lipids using the developed extraction protocol. The BUME protocol includes an initial one-phase extraction of plasma into 300 µl butanol:methanol (BUME) mixture (3:1) followed by two-phase extraction into 300 µl heptane:ethyl acetate (3:1) using 300 µl 1% acetic acid as buffer. The lipids investigated included the most abundant plasma lipid classes (e.g., cholesterol ester, free cholesterol, triacylglycerol, phosphatidylcholine, and sphingomyelin) as well as less abundant but biologically important lipid classes, including ceramide, diacylglycerol, and lyso-phospholipids. This novel method has been successfully implemented in our laboratory and is now used daily. We conclude that the fully automated, high-throughput BUME method can replace chloroform-based methods, saving both human and environmental resources. PMID:22645248

  13. Gas sensing in microplates with optodes: influence of oxygen exchange between sample, air, and plate material.

    PubMed

    Arain, Sarina; Weiss, Svenja; Heinzle, Elmar; John, Gernot T; Krause, Christian; Klimant, Ingo

    2005-05-01

    Microplates with integrated optical oxygen sensors are a new tool to study metabolic rates and enzyme activities. Precise measurements are possible only if oxygen exchange between the sample and the environment is known. In this study we quantify gas exchange in plastic microplates. Dissolved oxygen was detected using either an oxygen-sensitive film fixed at the bottom of each well or a needle-type sensor. The diffusion of oxygen into wells sealed with different foils, paraffin oil, and paraffin wax, respectively, was quantified. Although foil covers showed the lowest oxygen permeability, they include an inevitable gas phase between sample and sealing and are difficult to manage. The use of oil was found to be critical due to the extensive shaking caused by movement of the plates during measurements in microplate readers. Thus, paraffin wax was the choice material because it avoids convection of the sample and is easy to handle. Furthermore, without shaking, significant gradients in pO2 levels within a single well of a polystyrene microplate covered with paraffin oil were detected with the needle-type sensor. Higher pO2 levels were obtained near the surface of the sample as well as near the wall of the well. A significant diffusion of oxygen through the plastic plate material was found using plates based on polystyrene. Thus, the location of a sensor element within the well has an effect on the measured pO2 level. Using a sensor film fixed on the bottom of a well or using a dissolved pO2-sensitive indicator results in pO2 offset and in apparently lower respiration rates or enzyme activities. Oxygen diffusion through a polystyrene microplate was simulated for measurements without convection--that is, for samples without oxygen diffusion through the cover and for unshaken measurements using permeable sealings. This mathematical model allows for calculation of the correct kinetic parameters. PMID:15772950

  14. Determination of suvorexant in human plasma using 96-well liquid-liquid extraction and HPLC with tandem mass spectrometric detection.

    PubMed

    Breidinger, S A; Simpson, R C; Mangin, E; Woolf, E J

    2015-10-01

    A method, using liquid chromatography with tandem mass spectrometric detection (LC-MS/MS), was developed for the determination of suvorexant (MK-4305, Belsomra(®)), a selective dual orexin receptor antagonist for the treatment insomnia, in human plasma over the concentration range of 1-1000ng/mL. Stable isotope labeled (13)C(2)H3-suvorexant was used as an internal standard. The sample preparation procedure utilized liquid-liquid extraction, in the 96-well format, of a 100μL plasma sample with methyl t-butyl ether. The compounds were chromatographed under isocratic conditions on a Waters dC18 (50×2.1mm, 3μm) column with a mobile phase consisting of 30/70 (v/v %) 10mM ammonium formate, pH3/acetonitrile at a flow rate of 0.3mL/min. Multiple reaction monitoring of the precursor-to-product ion pairs for suvorexant (m/z 451→186) and (13)C(2)H3-suvorexant (m/z 455→190) on an Applied Biosystems API 4000 tandem mass spectrometer was used for quantitation. Intraday assay precision, assessed in six different lots of control plasma, was within 10% CV at all concentrations, while assay accuracy ranged from 95.6 to 105.0% of nominal. Quality control (QC) samples in plasma were stored at -20°C. Initial within day analysis of QCs after one freeze-thaw cycle showed accuracy within 9.5% of nominal with precision (CV) of 6.7% or less. The plasma QC samples were demonstrated to be stable for up to 25 months at -20°C. The method described has been used to support clinical studies during Phase I through III of clinical development. PMID:26343269

  15. Fluorescent protein-based cellular assays analyzed by laser-scanning microplate cytometry in 1536-well plate format.

    PubMed

    Auld, Douglas S; Johnson, Ronald L; Zhang, Ya-qin; Veith, Henrike; Jadhav, Ajit; Yasgar, Adam; Simeonov, Anton; Zheng, Wei; Martinez, Elisabeth D; Westwick, John K; Austin, Christopher P; Inglese, James

    2006-01-01

    Microtiter plate readers have evolved from photomultiplier and charged-coupled device-based readers, where a population-averaged signal is detected from each well, to microscope-based imaging systems, where cellular characteristics from individual cells are measured. For these systems, speed and ease of data analysis are inversely proportional to the amount of data collected from each well. Microplate laser cytometry is a technology compatible with a 1536-well plate format and capable of population distribution analysis. Microplate cytometers such as the Acumen Explorer can monitor up to four fluorescent signals from single objects in microtiter plates with densities as high as 1536 wells. These instruments can measure changes in fluorescent protein expression, cell shape, or simple cellular redistribution events such as cytoplasmic to nuclear translocation. To develop high-throughput screening applications using laser-scanning microplate cytometry, we used green fluorescent protein- and yellow fluorescent protein-expressing cell lines designed to measure diverse biological functions such as nuclear translocation, epigenetic signaling, and G protein-coupled receptor activation. This chapter illustrates the application of microplate laser cytometry to these assays in a manner that is suitable for screening large compound collections in high throughput. PMID:17110211

  16. Review of lignocellulolytic enzyme activity analyses and scale-down to microplate-based assays.

    PubMed

    Mansour, A A; Da Costa, A; Arnaud, T; Lu-Chau, T A; Fdz-Polanco, Maria; Moreira, M T; Cacho Rivero, J A

    2016-04-01

    With the increasing use of enzymes in environmental applications, there is a need for analytical methods adapted to large factorial experiments. Existing reference methods are chemical and labor intensive and unsuitable to analyze in parallel a large number of samples. Based on an extensive literature review and on experimental results, this work compares reference and microplate adapted methods to define the most adequate filter paper, carboxymethylcellulase, β-glucosidase and xylanase activity tests. In the adapted methods, the total reaction volume was reduced from 2.2-24.5 mL to 0.21-0.24 mL. Statistical analysis of the activities measured on enzyme mixtures by applying the 96-well plate reduced methods showed that they were not significantly different to the activities obtained with reference tests. PMID:26838452

  17. Reader Pals.

    ERIC Educational Resources Information Center

    Ayres, Linda; Wainess, Didi

    1989-01-01

    Reader Pals is a cross-age, whole language, independent, daily reading program designed to supplement existing reading curricula and to enrich daily lessons. A younger child is paired with an older child for a year. Program planning and implementation are discussed, and activities based on a dinosaur theme are suggested. (IAH)

  18. 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 PRESENTED ON…

  19. 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

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

    PubMed

    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

  1. 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.

  2. Microplate-Based Characterization of Protein-Phosphoinositide Binding Interactions Using a Synthetic Biotinylated Headgroup Analogue

    PubMed Central

    Gong, Denghuang; Smith, Matthew D.; Manna, Debasis; Bostic, Heidi E.; Cho, Wonhwa; Best, Michael D.

    2009-01-01

    Membrane lipids act as important regulators of a litany of important physiological and pathophysiological events. Many of them act as site-specific ligands for cytosolic proteins in binding events that recruit receptors to the cell surface and control both protein function and subcellular localization. Phosphatidylinositol phosphates (PIPns) are a family of signaling lipids that regulate numerous cellular processes by interacting with a myriad of protein binding modules. Characterization of PIPn-binding proteins has been hampered by the lack of a rapid and convenient quantitative assay. Herein, microplate-based detection is presented as an effective approach to characterizing protein-PIPn binding interactions at the molecular level. With this assay, the binding of proteins to isolated PIPn headgroups is detected with high sensitivity using a platform that is amenable to high-throughput screening. In the studies described herein, biotinylated PI-(4,5)-P2 headgroup analogue 1 was designed, synthesized and immobilized onto 96-well streptavidin-coated microplates to study receptor binding. This assay was used to characterize the binding of the PH domain of β-spectrin to this headgroup. The high affinity interaction that was detected for surface association (Kd, surf = 6 nM ±3), demonstrates that receptor binding modules can form high affinity interactions with lipid headgroups outside of a membrane environment. The results also indicate the feasibility of the assay for rapid characterization of PIPn-binding proteins as well as the promise for high-throughput analysis of protein-PIPn binding interactions. Finally, this assay was also employed to characterize the inhibition of the binding of receptors to the PIPn-derivatized microplates using solution phase competitors. This showcases the viability of this assay for rapid screening of inhibitors of PIPn-binding proteins. PMID:19182890

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

    PubMed

    Gülden, Michael; Schreiner, Jeannine; Seibert, Hasso

    2015-06-01

    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. PMID:24291469

  4. Architecture of the Northwest Andean Microplates

    NASA Astrophysics Data System (ADS)

    Potts, L. V.; Hernandez, O.; von Frese, R. R.; Schmidt, M.

    2005-05-01

    Recently revised models on global plate boundary zones show that the North Andes microplate includes a wide distribution of seismicity, volcanic events, active faulting and extreme topography. The current description of the north Andean microplate boundaries is interpreted from a variety of geological and geophysical models including volcanism and seismicity with variable confidence levels. The poorly understood complex structure and geometry of plate boundaries limits the ability of current physical models to predict neotectonic and other effects including intra-plate lithospheric stresses and strain. Together with local surface gravity and topography data, a variety of available space geodetic sensors have substantially improved the modeling of the lithosphere for analyzing subsurface mass dynamics. They include the GPS-derived 3-D crustal velocities, high resolution (90-m) topography, seismic surveys and high resolution gravity models derived from integrated satellite (e.g., CHAMP and GRACE, 200-km resolution) and terrestrial observations (up to ~5 km resolution). Spherical wavelets is a modern tool for a multi-resolution representation of spatially heterogenously distributed gravity (consistent with the generalized boundary value problem) and for topography datasets with the distinct ability to enhance localized signals. Analysis of multi-resolution gravity and topography models combined with GPS velocities provide a unique opportunity to characterize the structure, isostatic conditions, mass dynamics, and intra-plate deformations of the North Andes microplate.

  5. Determination of N-species in soil extracts using microplate techniques.

    PubMed

    Shand, Charles A; Williams, Berwyn L; Coutts, Grace

    2008-01-15

    Colourimetric methods for the determination of NO(3)(-), NH(4)(+) and total N in water extracts of soils using 96-well microplate techniques are described. Nitrate was determined by azo dye formation after reduction to NO(2)(-) using a solution of hydrazine sulphate. Ammonium in the soil extracts was purified and concentrated by diffusion as NH(3) from small volumes (750microL) of extract treated with MgO into a H(2)SO(4) collector using a double-plate, MicroResp method and subsequently determined by the Berthelot reaction. For the determination of total N, samples were oxidised with K(2)S(2)O(8) at 110 degrees C in a 96x1.1mL polytetrafluoroethylene block with a lid that closed individual wells. The oxidised solutions were transferred to standard plates for colourimetric analysis of NO(3)(-). The recovery of N, measured as NO(3)(-), from NH(4)NO(3) and a range of organic-N compounds was >95%. The limits of quantitation of the colourimetic assays were 0.020mgNL(-1) for NO(3)(-) and 0.051mgNL(-1) for NH(4)(+). The methods were tested on water extracts derived from a range of 10 nutrient poor soils from Scotland. There were acceptable linear correlations between the results obtained by established methods. For soil extracts analysed by the microplate method, the relationship for NO(3)(-) was 1.03x result from ion chromatography+0.0055 (R(2)=0.9961); for NH(4)(+) determined by the microplate method, the relationship was 0.9696xresult from a discrete analyser-0.0169 (R(2)=0.9757) and for total N determined by oxidation in the PTFE microplate the relationship was 0.9435xresult obtained by combustion+0.0489 (R(2)=0.9743). Purification of the NH(4)(+) in water extracts from the 10 different soils by the diffusion method did not result in any systematic difference (paired t-test, p=0.05) between measured concentration values determined before and after diffusion. PMID:18371688

  6. High-throughput genotyping using horizontal polyacrylamide gels with wells arranged for microplate array diagonal gel electrophoresis (MADGE).

    PubMed

    Day, I N; Humphries, S E; Richards, S; Norton, D; Reid, M

    1995-11-01

    Genotyping (typing of genetic variation) typically involves PCR followed by an allele-specific oligonucleotide-binding assay, restriction enzyme digest or direct check of the outcome of a PCR designed to distinguish genotype. Electrophoresis can resolve "bound" from "free" oligonucleotide, as well as resolve PCR fragments and digests, but it is traditionally regarded as cumbersome and laborious in comparison with solution assays. Here we describe simple horizontal polyacrylamide gels which can receive a 96-well array of samples directly, which can be stacked in tanks and which are bound to a robust support of glass. The line of electrophoresis is on a 71.6 degree diagonal relative to the columns of the array (microplate array diagonal gel electrophoresis [MADGE]). Several thousand reactions can conveniently be analyzed in a shoebox-sized apparatus in a couple of hours. High resolution is achieved in the range of 20-1000 bp, information processing is simplified and automation is possible. PMID:8588924

  7. Photonic crystal biosensor microplates with integrated fluid networks for high throughput applications in drug discovery

    NASA Astrophysics Data System (ADS)

    Choi, Charles J.; Chan, Leo L.; Pineda, Maria F.; Cunningham, Brian T.

    2007-09-01

    Assays used in pharmaceutical research require a system that can not only detect biochemical interactions with high sensitivity, but that can also perform many measurements in parallel while consuming low volumes of reagents. While nearly all label-free biosensor transducers to date have been interfaced with a flow channel, the liquid handling system is typically aligned and bonded to the transducer for supplying analytes to only a few sensors in parallel. In this presentation, we describe a fabrication approach for photonic crystal biosensors that utilizes nanoreplica molding to produce a network of sensors that are automatically self-aligned with a microfluidic network in a single process step. The sensor/fluid network is inexpensively produced on large surface areas upon flexible plastic substrates, allowing the device to be incorporated into standard format 96-well microplates. A simple flow scheme using hydrostatic pressure applied through a single control point enables immobilization of capture ligands upon a large number of sensors with 220 nL of reagent, and subsequent exposure of the sensors to test samples. A high resolution imaging detection instrument is capable of monitoring the binding within parallel channels at rates compatible with determining kinetic binding constants between the immobilized ligands and the analytes. The first implementation of this system is capable of monitoring the kinetic interactions of 11 flow channels at once, and a total of 88 channels within an integrated biosensor microplate in rapid succession. The system was initially tested to characterize the interaction between sets of proteins with known binding behavior.

  8. 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

  9. Development of a novel 96-well format for liquid-liquid microextraction and its application in the HPLC analysis of biological samples.

    PubMed

    Borijihan, Guirong; Li, Youxin; Gao, Jianguo; Bao, James J

    2014-05-01

    A novel 96-well liquid-liquid microextraction system combined with modern HPLC was developed and used for the simultaneous analysis of 96 biological samples. The system made use of hollow fibers, a 96-well plate, and a plastic base with a center hole and a side hole. One end of the hollow fiber was sealed, while the other end was attached to one of the holes positioned at the center for the plastic base. The needle was inserted into the liquid from inside or outside of the hollow fiber through the center or the side holes, respectively. The system was tested with plasma samples containing three compounds, acidic indomethacin, neutral dexamethasone, and basic propafenone. Some parameters, such as the kind and dimension of hollow fiber, pH and salt concentration of the donor phase, the selection of organic solvent for the acceptor phase, and the extraction time were investigated. Under the optimization conditions, the Log D and drug concentration of indomethacin, dexamethasone, and propafenone in plasma and urine samples were analyzed. Then, the methodology was validated. The results demonstrated that ng/mL levels could be exactly and rapidly analyzed by our system, which was equipped with an auto-injection sampler, making sample analysis more convenient. PMID:24574156

  10. Miniaturized plate readers for low-cost, high-throughput phenotypic screening.

    PubMed

    Jensen, Paul A; Dougherty, Bonnie V; Moutinho, Thomas J; Papin, Jason A

    2015-02-01

    We present a miniaturized plate reader for measuring optical density in 96-well plates. Our standalone reader fits in most incubators, environmental chambers, or biological containment suites, allowing users to leverage their existing laboratory infrastructure. The device contains no moving parts, allowing an entire 96-well plate to be read several times per second. We demonstrate how the fast sampling rate allows our reader to detect small changes in optical density, even when the device is placed in a shaking incubator. A wireless communication module allows remote monitoring of multiple devices in real time. These features allow easy assembly of multiple readers to create a scalable, accurate solution for high-throughput phenotypic screening. PMID:25366331

  11. Miniaturized Plate Readers for Low-Cost, High-Throughput Phenotypic Screening

    PubMed Central

    Jensen, Paul A.; Dougherty, Bonnie V.; Moutinho, Thomas J.

    2015-01-01

    We present a miniaturized plate reader for measuring optical density in 96-well plates. Our standalone reader fits in most incubators, environmental chambers, or biological containment suites, allowing users to leverage their existing laboratory infrastructure. The device contains no moving parts, allowing an entire 96-well plate to be read several times per second. We demonstrate how the fast sampling rate allows our reader to detect small changes in optical density, even when the device is placed in a shaking incubator. A wireless communication module allows remote monitoring of multiple devices in real time. These features allow easy assembly of multiple readers to create a scalable, accurate solution for high-throughput phenotypic screening. PMID:25366331

  12. Collection, storage, and filtration of in vivo study samples using 96-well filter plates to facilitate automated sample preparation and LC/MS/MS analysis.

    PubMed

    Berna, M; Murphy, A T; Wilken, B; Ackermann, B

    2002-03-01

    The benefits of high-throughput bioanalysis within the pharmaceutical industry are well established. One of the most significant bottlenecks in bioanalysis is transferring in vivo-generated study samples from their collection tubes during sample preparation and extraction. In most cases, the plasma samples must be stored frozen prior to analysis, and the freeze/thaw (F/T) process introduces thrombin clots that are capable of plugging pipets and automated liquid-transfer systems. A new approach to dealing with this problem involves the use of Ansys Captiva 96-well 20-microm polypropylene filter plates to collect, store frozen, and filter plasma samples prior to bioanalysis. The samples are collected from the test subjects, and the corresponding plasma samples are placed directly into the wells of the filter plate. Two Duoseal (patent pending) covers are used to seal the top and bottom of the plate, and the plate is stored at down to -70 degrees C. Prior to sample analysis, the seals are removed and the plate is placed in a 96-well SPE manifold. As the plasma thaws, it passes (by gravity or mild vacuum) through the polypropylene filter into a 96-well collection plate. A multichannel pipet or automated liquid-transfer system is used to transfer sample aliquots without fear of plugging. A significant advantage of this approach is that, unlike other methods, issues related to incomplete pipetting are virtually eliminated. The entire process is rapid since thawing and filtering take place simultaneously, and if a second F/T cycle is required for reanalysis, it is not necessary to refilter the samples (additional clotting was not observed after three F/T cycles). This technique was tested using monkey, rat, and dog plasma and sodium heparin and EDTA anticoagulants. To assess the possibility of nonspecific binding to the polypropylene filter, a variety of drug candidates from diverse drug classes were studied. Validation data generated for two Lilly compounds from distinct

  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. PMID:21069472

  14. Rapid microplate high-throughput methodology for assessment of Folin-Ciocalteu reducing capacity.

    PubMed

    Magalhães, Luís M; Santos, Fernando; Segundo, Marcela A; Reis, Salette; Lima, José L F C

    2010-12-15

    In the present work, a rapid and high-throughput Folin-Ciocalteu (F-C) reducing capacity assay adapted to routine/screening analysis was developed. In order to attain a fast F-C reducing kinetic reaction, the reaction conditions of the classical time-consuming F-C assay were modified and the influence of alkali and F-C reagent concentration was evaluated using gallic acid as standard. The proposed method was performed in a 96-well microplate format and it was applied to several phenolic compounds and food products (wines, beers, infusions and juices) providing F-C reducing capacity results after 3 min of reaction similar to those obtained by the time-consuming (120 min) conventional method. The additive and synergistic effect of reducing nonphenolic compounds usually found in food samples was also investigated. Ascorbic acid and ferrous sulfate provided an additive effect, while for fructose, glucose and sodium sulfite a synergistic effect was obtained. The detection limit was 0.25 mg L(-1) (as gallic acid) and the repeatability was <1.6% (n=12). PMID:21111158

  15. What Can Readers Read after Graded Readers?

    ERIC Educational Resources Information Center

    McQuillan, Jeff

    2016-01-01

    Nation (2014) concluded that most of the vocabulary one needs to read challenging texts in English can be acquired incidentally through voluminous reading. This study examines possible texts that second language (L2) readers can use to move from controlled-vocabulary materials such as graded readers, which go up through approximately the…

  16. Nurturing Lifelong Readers

    ERIC Educational Resources Information Center

    Judkins, Gerri

    2010-01-01

    In this paper the author relates how the thing he enjoys most about his job as a school librarian is nurturing lifelong readers, in particular readers of children's literature. In talking about nurturing lifelong readers and, referring to students, staff and school librarians, the author discusses: (1) The Southwell Library Programme; (2) The Lit…

  17. 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.

  18. Identification of photosynthesis inhibitors of pelagic marine algae using 96-well plate microfractionation for enhanced throughput in effect-directed analysis.

    PubMed

    Booij, Petra; Vethaak, A Dick; Leonards, Pim E G; Sjollema, Sascha B; Kool, Jeroen; de Voogt, Pim; Lamoree, Marja H

    2014-07-15

    Because of large-scale production and use of an increasing diversity of chemicals in modern society, estuarine and coastal waters may be contaminated with numerous substances. Some of these compounds have the potential to affect microalgae at the base of the pelagic food chain. Therefore, we identified the main chemical stressors that negatively affect the effective photosystem II efficiency (ϕPSII) in marine microalgae of the Dutch estuarine and coastal waters. An enhanced effect-directed analysis (EDA) was carried out by combining reversed-phase ultra performance liquid chromatography fractionation of extracts from passive samplers, followed by effect assessment using the pulse amplitude modulation fluorometry assay and chemical analysis of biologically active fractions using high-resolution mass spectrometry. This study focuses on a novel microfractionation technique using 96-well plates to enhance throughput in EDA, structure elucidation, and the analytical and effect confirmation of the compounds that are identified. Although there are numerous unknown compounds present in estuarine and coastal waters, our EDA study shows that atrazine, diuron, irgarol, isoproturon, terbutryn, and terbutylazine are the main contributors to the observed effect on the ϕPSII of marine microalgae. PMID:24926900

  19. High throughput adjustable 96-well plate assay for androgen receptor binding: a practical approach for EDC screening using the chimpanzee AR.

    PubMed

    Hartig, P C; Cardon, M C; Blystone, C R; Gray, L E; Wilson, V S

    2008-09-26

    The issue as to whether natural and man-made chemicals interfere with endocrine function has raised concerns. This interference could be biologically significant even at very low doses if the chemicals interact deleteriously with hormone receptors at low concentrations. Therefore, the United States Environmental Protection Agency (USEPA) Office of Coordination and Policy (OSCP) requested that a nonhuman mammalian androgen receptor binding assay be developed for possible use in their Endocrine Disruptor Screening Program (EDSP). Ideally, this assay would be high throughput, not use animals as a source of receptor protein, easily deployed throughout the scientific community, utilize reagents available to both the public and private sector, and have the potential for future automation. We developed a highly modified 96-well plate assay which meets these criteria. It employs a baculovirus expressed recombinant primate androgen receptor which is publically available and exploits the unique ability of some mammalian androgen receptors to remain biologically active after guanidine hydrochloride (GdnHCl) solubilization. This GdnHCl treated receptor remains soluble and requires no additional purification prior to use. We provide a very detailed description of the assay protocol itself, and similarly detailed method for producing and solubilizing the receptor. PMID:18691642

  20. Characterization of multiple platelet activation pathways in patients with bleeding as a high-throughput screening option: use of 96-well Optimul assay

    PubMed Central

    Lordkipanidzé, Marie; Lowe, Gillian C.; Kirkby, Nicholas S.; Chan, Melissa V.; Lundberg, Martina H.; Morgan, Neil V.; Bem, Danai; Nisar, Shaista P.; Leo, Vincenzo C.; Jones, Matthew L.; Mundell, Stuart J.; Daly, Martina E.; Mumford, Andrew D.; Warner, Timothy D.; Watson, Steve P.; Watson, Steve P.; Mumford, Andrew D.; Mundell, Stuart J.; Gissen, Paul; Daly, Martina E.; Lester, Will; Clark, Justin; Williams, Mike; Motwani, Jayashree; Marshall, Dianne; Nyatanga, Priscilla; Mann, Pat; Kirwan, Julie; Wilde, Jonathan; Dunkley, Tracey; Greenway, April; Makris, Michael; Pavord, Sue; Dattani, Rashesh; Grimley, Gerry Dolan Charlotte; Stokley, Simone; Astwood, Emma; Chang, Cherry; Foros, Merri; Trower, Linda; Thachil, Jecko; Hay, Charlie; Pike, Gill; Will, Andrew; Grainger, John; Foulkes, Matt; Fareh, Mona; Talks, Kate; Biss, Tina; Kesteven, Patrick; Hanley, John; Vowles, Julie; Basey, Lesley; Barnes, Michelle; Collins, Peter; Rayment, Rachel; Alikhan, Raza; Morris, Ana Guerrero Rebecca; Mansell, Dianne; Toh, Cheng Hock; Martlew, Vanessa; Murphy, Elaine; Lachmann, Robin; Rose, Peter; Chapman, Oliver; Lokare, Anand; Marshall, Kathryn; Khan, Naseem; Keeling, David; Giangrande, Paul; Austin, Steve; Bevan, David; Alamelu, Jayanthi

    2014-01-01

    Up to 1% of the population have mild bleeding disorders, but these remain poorly characterized, particularly with regard to the roles of platelets. We have compared the usefulness of Optimul, a 96-well plate-based assay of 7 distinct pathways of platelet activation to characterize inherited platelet defects in comparison with light transmission aggregometry (LTA). Using Optimul and LTA, concentration-response curves were generated for arachidonic acid, ADP, collagen, epinephrine, Thrombin receptor activating-peptide, U46619, and ristocetin in samples from (1) healthy volunteers (n = 50), (2) healthy volunteers treated with antiplatelet agents in vitro (n = 10), and (3) patients with bleeding of unknown origin (n = 65). The assays gave concordant results in 82% of cases (κ = 0.62, P < .0001). Normal platelet function results were particularly predictive (sensitivity, 94%; negative predictive value, 91%), whereas a positive result was not always substantiated by LTA (specificity, 67%; positive predictive value, 77%). The Optimul assay was significantly more sensitive at characterizing defects in the thromboxane pathway, which presented with normal responses with LTA. The Optimul assay is sensitive to mild platelet defects, could be used as a rapid screening assay in patients presenting with bleeding symptoms, and detects changes in platelet function more readily than LTA. This trial was registered at www.isrctn.org as #ISRCTN 77951167. PMID:24408324

  1. Characterization of multiple platelet activation pathways in patients with bleeding as a high-throughput screening option: use of 96-well Optimul assay.

    PubMed

    Lordkipanidzé, Marie; Lowe, Gillian C; Kirkby, Nicholas S; Chan, Melissa V; Lundberg, Martina H; Morgan, Neil V; Bem, Danai; Nisar, Shaista P; Leo, Vincenzo C; Jones, Matthew L; Mundell, Stuart J; Daly, Martina E; Mumford, Andrew D; Warner, Timothy D; Watson, Steve P

    2014-02-20

    Up to 1% of the population have mild bleeding disorders, but these remain poorly characterized, particularly with regard to the roles of platelets. We have compared the usefulness of Optimul, a 96-well plate-based assay of 7 distinct pathways of platelet activation to characterize inherited platelet defects in comparison with light transmission aggregometry (LTA). Using Optimul and LTA, concentration-response curves were generated for arachidonic acid, ADP, collagen, epinephrine, Thrombin receptor activating-peptide, U46619, and ristocetin in samples from (1) healthy volunteers (n = 50), (2) healthy volunteers treated with antiplatelet agents in vitro (n = 10), and (3) patients with bleeding of unknown origin (n = 65). The assays gave concordant results in 82% of cases (κ = 0.62, P < .0001). Normal platelet function results were particularly predictive (sensitivity, 94%; negative predictive value, 91%), whereas a positive result was not always substantiated by LTA (specificity, 67%; positive predictive value, 77%). The Optimul assay was significantly more sensitive at characterizing defects in the thromboxane pathway, which presented with normal responses with LTA. The Optimul assay is sensitive to mild platelet defects, could be used as a rapid screening assay in patients presenting with bleeding symptoms, and detects changes in platelet function more readily than LTA. This trial was registered at www.isrctn.org as #ISRCTN 77951167. PMID:24408324

  2. 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. PMID:9448849

  3. Determination of the HIV integrase inhibitor, MK-0518 (raltegravir), in human plasma using 96-well liquid-liquid extraction and HPLC-MS/MS.

    PubMed

    Merschman, S A; Vallano, P T; Wenning, L A; Matuszewski, B K; Woolf, E J

    2007-09-15

    An HPLC-MS/MS method was developed for the determination of MK-0518 (raltegravir), an HIV integrase inhibitor, in human plasma over the concentration range of 2-1000 ng/mL. Stable isotope labeled (13)C(6)-MK-0518 was used as an internal standard. The sample preparation procedure utilized liquid-liquid extraction with hexane:methylene chloride in the 96-well format with a 200 microL plasma sample size. The compounds were chromatographed on an Ace C(18) (50 x 3.0 mm, 3 microm, titanium frits) column with 42.5/57.5 (v/v %) 0.1mM EDTA in 0.1% formic acid/methanol mobile phase at a flow rate of 0.5 mL/min. Multiple reaction monitoring of the precursor-to-product ion pairs for MK-0518 (m/z 445-->109) and (13)C(6)-MK-0518 (m/z 451-->367) on an Applied Biosystem API 4000 HPLC-MS/MS was used for quantitation. Intraday precision of standard curve concentrations in five different lots of control plasma was within 3.2%, while accuracy ranged from 94.8 to 106.8%. The mean extraction recovery of spiked plasma samples was 87%. Quality control (QC) samples were stored at -20 degrees C. Initial within day analysis showed QC accuracy within 7.5% of nominal with precision of 3.1% or less. The plasma QC samples were demonstrated to be stable for up to 23 months at -20 degrees C. The method described has been used to support over 18 clinical studies during Phase I through III of clinical development. PMID:17644453

  4. The origin of the Iberian microplate high topography

    NASA Astrophysics Data System (ADS)

    Quintana, L.; Pulgar, J. A.; Alonso, J. L.; Fernandez Viejo, G.

    2013-12-01

    The Iberian microplate is about 1100 km width, 3500 km long, and experienced contraction during the Cenozoic convergence of the Eurasian and African plates. In this process two main mountain ranges were created in the boundaries of the microplate: the Cantabrian-Pyrenean range in the north and the Betic-Rif orogenic system in the south, with a maximun height of 3479 m over sea level. However, in the interior of the microplate there is also several ranges and plateaus, with maximun heights of 2592 m. The origin of these interior high topography is controversial and has been explained by means of several procesess as: a) transference of displacement from the south border of the microplate, b) lithospheric folding, or c) more complex deep mantle related processes. In this work we investigate the relation between the northern border range of the microplate and some Iberian interior reliefs as: a) the Spanish Central System range, located in the central part of the microplate, with peaks over 2500 m, and b) the North-Iberian meseta, situated between these two ranges, and with average heights of 750 m. In order to solve this relation we have built a crustal-scale cross-section, through the northern half of the Iberian microplate, by using field geological mapping, subsoil information and deep geophysical data. The section acroos the central part of the Cantabrian range shows the well-known Iberian microplate continental subduction beneath the Eurasian plate, with a prominent crustal root that reaches up to 45 km depth. In this section an important difference between the upper and lower crust shortening values has been found. Thus, the upper crust has been shortened 97 km while the lower crust 122 km. The section across de Central System range shows a slightly thickened crust. In this range also exist a difference in the shortening values between upper and lower crust, but in the contrary sense than the Cantabrian range. Thus, the Central System range has been shortened

  5. 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. PMID:27581492

  6. A single-islet microplate assay to measure mouse and human islet insulin secretion.

    PubMed

    Truchan, Nathan A; Brar, Harpreet K; Gallagher, Shannon J; Neuman, Joshua C; Kimple, Michelle E

    2015-01-01

    One complication to comparing β-cell function among islet preparations, whether from genetically identical or diverse animals or human organ donors, is the number of islets required per assay. Islet numbers can be limiting, meaning that fewer conditions can be tested; other islet measurements must be excluded; or islets must be pooled from multiple animals/donors for each experiment. Furthermore, pooling islets negates the possibility of performing single-islet comparisons. Our aim was to validate a 96-well plate-based single islet insulin secretion assay that would be as robust as previously published methods to quantify glucose-stimulated insulin secretion from mouse and human islets. First, we tested our new assay using mouse islets, showing robust stimulation of insulin secretion 24 or 48 h after islet isolation. Next, we utilized the assay to quantify mouse islet function on an individual islet basis, measurements that would not be possible with the standard pooled islet assay methods. Next, we validated our new assay using human islets obtained from the Integrated Islet Distribution Program (IIDP). Human islets are known to have widely varying insulin secretion capacity, and using our new assay we reveal biologically relevant factors that are significantly correlated with human islet function, whether displayed as maximal insulin secretion response or fold-stimulation of insulin secretion. Overall, our results suggest this new microplate assay will be a useful tool for many laboratories, expert or not in islet techniques, to be able to precisely quantify islet insulin secretion from their models of interest. PMID:26452321

  7. 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, especially…

  8. 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…

  9. 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)

  10. 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…

  11. Intermediate Cambodian Reader.

    ERIC Educational Resources Information Center

    Huffman, Franklin E., Ed.; Proum, Im, Ed.

    This book is a sequel to the "Cambodian System of Writing and Beginning Reader." It is intended to serve as an intermediate reader to develop the student's ability to the point of reading Cambodian texts with the aid of a dictionary. Part One of the book consists of 37 readings, graded in length and difficulty, and selected to provide a wide range…

  12. The Self Actualized Reader.

    ERIC Educational Resources Information Center

    Marino, Michael; Moylan, Mary Elizabeth

    A study examined the commonalities that "voracious" readers share, and how their experiences can guide parents, teachers, and librarians in assisting children to become self-actualized readers. Subjects, 25 adults ranging in age from 20 to 67 years, completed a questionnaire concerning their reading histories and habits. Respondents varied in…

  13. 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%.

  14. 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

    2015-02-01

    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%. PMID:25448978

  15. 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. PMID:27241123

  16. 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.

  17. 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. PMID:14558996

  18. Quantitative Microplate-Based Respirometry with Correction for Oxygen Diffusion

    PubMed Central

    2009-01-01

    Respirometry using modified cell culture microplates offers an increase in throughput and a decrease in biological material required for each assay. Plate based respirometers are susceptible to a range of diffusion phenomena; as O2 is consumed by the specimen, atmospheric O2 leaks into the measurement volume. Oxygen also dissolves in and diffuses passively through the polystyrene commonly used as a microplate material. Consequently the walls of such respirometer chambers are not just permeable to O2 but also store substantial amounts of gas. O2 flux between the walls and the measurement volume biases the measured oxygen consumption rate depending on the actual [O2] gradient. We describe a compartment model-based correction algorithm to deconvolute the biological oxygen consumption rate from the measured [O2]. We optimize the algorithm to work with the Seahorse XF24 extracellular flux analyzer. The correction algorithm is biologically validated using mouse cortical synaptosomes and liver mitochondria attached to XF24 V7 cell culture microplates, and by comparison to classical Clark electrode oxygraph measurements. The algorithm increases the useful range of oxygen consumption rates, the temporal resolution, and durations of measurements. The algorithm is presented in a general format and is therefore applicable to other respirometer systems. PMID:19555051

  19. 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.

  20. Crocin bleaching antioxidant assay revisited: application to microplate to analyse antioxidant and pro-oxidant activities.

    PubMed

    Prieto, M A; Vázquez, J A; Murado, M A

    2015-01-15

    The crocin bleaching assay (CBA) is a common method for evaluating the antioxidant activity of hydrosoluble samples. It is criticised due to its low reproducibility, problematic quantification of results, differences in reagent preparation, doubtful need for a preheating phase and sensitivity to factors such as temperature, pH, solvents and metals. Here, the critical points of the method were extensively revised, and a highly reproducible procedure for microplate readers redeveloped. The problems of using quantification procedures, disregarding kinetic considerations, are discussed in detail and a model is proposed for quantifying simultaneously anti- and pro-oxidant activities as function of concentration and time. Thus, the combined use of a reproducible procedure and robust mathematical modeling produced consistent and meaningful criteria for comparative characterization of any oxidation modifier, taking into account the dose-time-dependent behaviour. The method was verified by characterising several commercial antioxidants and some metal compounds using the parametric values of the proposed models. The activity of the tested antioxidants decreased in the order ETX>TR>PG>AA>TBHQ>BHA. Others, such as the lipophilic antioxidants of BHT and α-Tocopherol did not show any activity. Interference from metals were for Fe(2+), Fe(3+), Cd(2+), Ni(2+), Mg(2+), Zn(2+) and Sr(2+), slightly antioxidant for Cu(1+) and Cu(2+), and strongly antioxidant for Mn(2+). None of the tested metals showed a pro-oxidant activity. PMID:25148992

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. Hungarian Graded Reader.

    ERIC Educational Resources Information Center

    Mihalyfy, Ilona; Koski, Augustus A.

    This graded reader is intended to supplement a beginning course in Hungarian after the student has "developed control of about 700 lexical items and can manipulate with fluency much of the basic structure of Hungarian." (It may also be used in an intermediate course.) The 56 reading selections are varied in content and sequenced to develop reading…

  6. Indonesian Newspaper Reader.

    ERIC Educational Resources Information Center

    Harter, Joseph M.; And Others

    This reader consists of 250 selections of varying lengths. No real attempt has been made to grade the selections according to difficulty for the learner except that the initial selections are shorter and represent straight reporting of such uncomplicated events as arrivals and departures and thus are fairly simple in structure. Notes explaining…

  7. 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…

  8. 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." (RM)

  9. Parents and Beginning Readers.

    ERIC Educational Resources Information Center

    Mathews, Virginia H., Ed.

    This series of 12 articles for parents on how to help pre-schoolers and beginning readers at home was written for the National Reading Center by specialists in reading and early childhood education. The titles and authors are: "Getting Ready to Read" (E. Robert La Crosse), "Creating a Good Reading Climate at Home" (Mary Frances K. Johnson),…

  10. 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…

  11. For Reluctant Readers.

    ERIC Educational Resources Information Center

    Fidell, Jeannette A.; Horn, Valerie

    The eleven lesson plans outlined in this booklet are designed to stimulate and challenge both underachievers and reluctant readers by helping them learn to read "The New York Times." The lesson plans include the following: exercises in vocabulary enrichment; exercises for developing dictionary skills; topics for oral reports; topics for written…

  12. Miami Linguistic Readers

    ERIC Educational Resources Information Center

    Bannatyne, Alex

    1973-01-01

    The Miami Linguistic Readers, which are comprised of 21 paperbound books grouped in 15 levels, concentrate on sounds and structure of standard American English for teaching linguistically handicapped or disadvantaged children before and during the teaching of basic reading skills. (MC)

  13. The Strategic Reader.

    ERIC Educational Resources Information Center

    Devine, James T., Ed.; And Others

    1986-01-01

    To add a foundation to the growing excitement among educators about the central role they play in helping learners become strategic readers, the articles in this thematic journal provide insight into current reading theory and practice. Richard Telfer's article reviews research on strategic reading and clarifies what is meant by the phrase…

  14. 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.

  15. 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…

  16. Constraints on the Ionian microplate subduction beneath the Tyrrhenian basin

    NASA Astrophysics Data System (ADS)

    Pasquale, V.; Verdoya, M.; Chiozzi, P.

    2003-04-01

    The Tyrrhenian Basin is, from a geodynamical point of view, one of the most intriguing areas of the Central Mediterranean, being affected by still ongoing and not yet fully understood tectonic processes of lithosphere extension and subduction. Its high heat flux, albeit lower than that expected from classical models of plate cooling, together with other pieces of geophysical evidence, argues for a marginal-type basin of very young age. The abyssal plain of the basin is floored with oceanic crust and it is thermally perturbed by strong hydrothermal circulation. The stretched continental margins are affected by volcanic activity, which causes local thermal perturbations by fluid circulation and magma intrusion. The surface heat flux budget on the margins is well balanced only if one envisages an additional thermal contribution of a layer of magma underplating the crust and a lithosphere overlying an anomalously hot asthenosphere. In this paper we supply further constraints on the deformation processes affecting the Ionian microplate subducting beneath the southern part of the basin, on the basis of a joint analysis of the seismic activity and the thermal state. We determined the thermal structure in the descending plate by means of an analytical model taking into account how the slab would heat and the mantle wedge cool with time. A relation between the maximum depth of seismicity of several marginal basins and the thermal parameter of the descending slabs, which depends on the age of the subducted lithosphere and the vertical component of the convergence rate, has been used. The tectonic subsidence of about 6000 m evaluated for the oceanic domain of the Ionian microplate implies a heat flux of 40 mW/m2, which is in agreement with the observed values. By considering that the maximum depth of the seismic events within the slab, a thermal parameter of about 4500 km, was found. The corresponding assimilation and cooling times of the microplate are 13 and 100 My

  17. Biochemical characterization and immobilization of Erwinia carotovoral-asparaginase in a microplate for high-throughput biosensing of l-asparagine.

    PubMed

    Labrou, Nikolaos E; Muharram, Magdy Mohamed

    2016-10-01

    l-Asparaginases (l-ASNase, E.C. 3.5.1.1) catalyze the conversion of l-asparagine to l-aspartic acid and ammonia. In the present work, a new form of l-ASNase from a strain of Erwinia carotovora (EcaL-ASNase) was cloned, expressed in Escherichia coli as a soluble protein and characterized. The enzyme was purified to homogeneity by a single-step procedure comprising ion-exchange chromatography. The properties of the recombinant enzyme were investigated employing kinetic analysis and molecular modelling and the kinetic parameters (Km, kcat) were determined for a number of substrates. The enzyme was used to assemble a microplate-based biosensor that was used for the development of a simple assay for the determination of l-asparagine in biological samples. In this sensor, the enzyme was immobilized by crosslinking with glutaraldehyde and deposited into the well of a microplate in 96-well format. The sensing scheme was based on the colorimetric measurement of ammonia formation using the Nessler's reagent. This format is ideal for micro-volume applications and allows the use of the proposed biosensor in high-throughput applications for monitoring l-asparagine levels in serum and foods samples. Calibration curve was obtained for l-asparagine, with useful concentration range 10-200μΜ. The biosensor had a detection limit of 10μM for l-asparagine. The method's reproducibility was in the order of ±3-6% and l-asparagine mean recoveries were 101.5%. PMID:27542748

  18. 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…

  19. Cognitive Development in Early Readers.

    ERIC Educational Resources Information Center

    Briggs, Chari; Elkind, David

    Some studies of early readers are discussed. It is pointed out that study of early readers has relevance for practical and theoretical issues in psychology and education. Of interest in this document are the following questions: (1) Are there any special talents or traits distinguishing early from non-early readers? (2) Do children who read early…

  20. Oceanic microplate formation records the onset of India-Eurasia collision

    NASA Astrophysics Data System (ADS)

    Matthews, Kara J.; Dietmar Müller, R.; Sandwell, David T.

    2016-01-01

    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 ('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 seen 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 Mammerickx Microplate formation is linked with the India-Eurasia collision (initial 'soft' 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. 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, the 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 precise means of dating the onset of the India-Eurasia collision, and is completely independent of and complementary to timing

  1. 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.

  2. 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

  3. Use of a Fluorometric Imaging Plate Reader in high-throughput screening

    NASA Astrophysics Data System (ADS)

    Groebe, Duncan R.; Gopalakrishnan, Sujatha; Hahn, Holly; Warrior, Usha; Traphagen, Linda; Burns, David J.

    1999-04-01

    High-throughput screening (HTS) efforts at Abbott Laboratories have been greatly facilitated by the use of a Fluorometric Imaging Plate Reader. The FLIPR consists of an incubated cabinet with integrated 96-channel pipettor and fluorometer. An argon laser is used to excite fluorophores in a 96-well microtiter plate and the emitted fluorometer. An argon laser is used to excite fluorophores in a 96-well microtiter plate and the emitted fluorescence is imaged by a cooled CCD camera. The image data is downloaded from the camera and processed to average the signal form each well of the microtiter pate for each time point. The data is presented in real time on the computer screen, facilitating interpretation and trouble-shooting. In addition to fluorescence, the camera can also detect luminescence form firefly luciferase.

  4. 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. PMID:22464560

  5. 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...

  6. Thermoluminescence dosimeter reader

    SciTech Connect

    Miyake, S.; Miura, N.

    1984-10-30

    A thermoluminescence dosimeter reader having a heater for heating a thermoluminescence element, a light measuring circuit for measuring circuit for measuring the intensity of the thermoluminescence emanated from the element when it is heated and a display device for displaying the reading of the dosage of radiation to which the element is exposed according to the intensity of the thermoluminescence is provided with a dosage information inputting means which outputs an electric signal having a value representing a predetermined reference dosage of radiation, a calculating means for calculating a calibration constant which is the ratio between the value of the electric signal and the output value of the light measuring circuit which is the measured value of the dosage of radiation of a reference thermoluminescence element which is exposed to the predetermined reference dosage of radiation, and a memory means for memorizing the calibration constant.

  7. Development of a microplate-based, electrophoretic fluorescent protein kinase a assay: comparison with filter-binding and fluorescence polarization assay formats.

    PubMed

    Miick, Siobhan M; Jalali, Shila; Dwyer, Brian P; Havens, John; Thomas, Donald; Jimenez, Manuel A; Simpson, Mathew T; Zile, Betsy; Huss, Karen L; Campbell, Robert M

    2005-06-01

    A microplate-based electrophoretic assay has been developed for the serine/threonine kinase protein kinase A (PKA). The ElectroCapture PKA assay developed uses a positively charged, lissamine-rhodamine-labeled kemptide peptide substrate for the kinase reaction and Nanogen's ElectroCapture HTS Workstation and 384-well laminated membrane plates to electrophoretically separate the negatively charged phosphorylated peptide product from the kinase reaction mix. After the electrophoretic separation, the amount of rhodamine-labeled phosphopeptide product was quantified using a Tecan Ultra384 fluorescence reader. The ElectroCapture PKA assay was validated with both known PKA inhibitors and library compounds. The pK(iapp) results obtained in the ElectroCapture PKA assay were comparable to those generated with current radioactive filter-binding assay and antibody-based competitive fluorescence polarization PKA assay formats. PMID:15964934

  8. The Malaysian Class Reader Programme.

    ERIC Educational Resources Information Center

    Raj, Devinder; Hunt, Brain

    1990-01-01

    Briefly describes reading standards in Malaysian schools, outlines a class reader program, and provides samples of teaching files designed to help teachers implement the program. (one reference. (GLR)

  9. 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.

  10. Microplate spectroscopic methods for determination of the organophosphate soman.

    PubMed

    Prokofieva, Daria Stanislavovna; Voitenko, Natalia Gennadievna; Gustyleva, Lyudmila Konstantinovna; Babakov, Vladimir Nikolaevich; Savelieva, Elena Igorevna; Jenkins, Richard Owen; Goncharov, Nikolay Vasilievich

    2010-06-01

    Two microplate spectroscopic methods for determination of organophosphates, based on inhibition of acetylcholinesterase activity, have been elaborated and evaluated for determination of the chemical weapon agent soman. The principal difference between the methods is that one measures reaction substrate concentration (elaborated from Hestrin), while the other measures reaction product (elaborated from Ellman). The linear ranges of the two methods were found to be similar. Although the limit of quantification was lower for the Ellman method (110 pM), the sensitivity coefficient was in favor of the Hestrin method (1.55-fold higher). The effects of the main soman hydrolysis products were consistent for the two methods: both methylphosphonic acid and pinacolyl methylphosphonic acid did not inhibit acetylcholinesterase activity. The main components of decontaminating solutions showed differential effects: while monoethanolamine had no influence upon results obtained by either method, hydrogen peroxide interfered with the Ellman method at far lower concentrations than with the Hestrin method. In practical applications involving samples containing hydrogen peroxide, the method based on Hestrin should be regarded as much more specific for OP determination than the Ellman method. PMID:20411202

  11. Microplates based on liquid bridges between glass rods.

    PubMed

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

    2013-05-01

    Microplating that (i) does not necessitate complex or precise machinery to dispense small liquid volumes, (ii) enables fluorescent optical diagnosis, and (iii) permits simple analyte mixing mechanically is desirable. We advance here a novel approach that employs the formation of a liquid bridge held in place by capillary forces between glass rod tubes located parallel to each other. Experimental investigations made on liquid filling characteristics show conformance to theoretical notions. Analytical development showed the presence of regions of minimal uncertainty in the cross-sectional area of the liquid body arising from variations in the contact angle which permit consistent fluorescence measurements. Cyclical translation of the rods relative to each other, which cause rupture and reattachment of the liquid bridge, was found to engender good mixing. Strong linear trends were found in fluorescence signals relative to EGFP fluorophore concentration using standard and optical fiber (which offer targeted) excitation illumination. The open nature of liquid handling in the approach reported here and the positive results obtained portend the ability for development as integrated lab-on-a-chip devices. PMID:23484766

  12. Myoglobin microplate assay to evaluate prevention of protein peroxidation.

    PubMed

    Marques, Sara S; Magalhães, Luís M; Mota, Ana I P; Soares, Tânia R P; Korsak, Barbara; Reis, Salette; Segundo, Marcela A

    2015-10-10

    The current therapeutic strategies are based on the design of multifunctional drug candidates able to interact with various disease related targets. Drugs that have the ability to scavenge reactive oxygen species (ROS), beyond their main therapeutic action, may prevent the oxidative damage of biomolecules. Therefore, analytical approaches that monitor in a continuous mode the ability of drugs to counteract peroxidation of physiologically relevant biotargets are required. In the present work, a microplate spectrophotometric assay is proposed to evaluate the ability of selected cardiovascular drugs, including angiotensin-converting enzyme (ACE) inhibitors, β -blockers and statins to prevent protein peroxidation. Myoglobin, which is a heme protein, and peroxyl radicals generated from thermolysis of 2,2'-azo-bis(2-amidinopropane) dihydrochloride at 37 °C, pH 7.4 were selected as protein model and oxidative species, respectively. Myoglobin peroxidation was continuously monitored by the absorbance decrease at 409 nm and the ability of drugs to counteract protein oxidation was determined by the calculation of the area under the curve upon the myoglobin oxidation. Fluvastatin (AUC₅₀=12.5 ± 1.2 μM) and enalapril (AUC₅₀=15.2 ± 1.8 μM) showed high ability to prevent myoglobin peroxidation, providing even better efficiency than endogenous antioxidants such as reduced glutathione. Moreover, labetalol, enalapril and fluvastatin prevent the autoxidation of myoglobin, while glutathione showed a pro-oxidant effect. PMID:26093510

  13. Top One Hundred Kids' Books: Readers Talking to Readers.

    ERIC Educational Resources Information Center

    Meyers, Elaine; Kist, Marijo; Sugden, Sarah

    1999-01-01

    Describes how a committee of children's librarians created a booklist that allowed young readers to talk to other readers through published book annotations. Discusses development, promotion, and judging of a contest that encouraged children to submit new ideas and resulted in a list of entries from children at 55 schools. Includes sample winning…

  14. 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…

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

    PubMed Central

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

    2016-01-01

    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. PMID:26832984

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

    NASA Astrophysics Data System (ADS)

    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.

  17. 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.…

  18. Fostering Fluent Readers and Writers.

    ERIC Educational Resources Information Center

    Goldsmith, E.

    In recent characterizations of reading and writing, the distinctions melt into each other so that one definition serves for both: both are the creation of meaning. In the act of creating meaning, readers become writers and writers become readers. While reading is primarily receptive and writing is primarily productive, fluency in reading is very…

  19. 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 interpretation of the subject…

  20. First hydrothermal active vent discovered on the Galapagos Microplate

    NASA Astrophysics Data System (ADS)

    Tao, C.; Li, H.; Wu, G.; Su, X.; Zhang, G.; Chinese DY115-21 Leg 3 Scientific Party

    2011-12-01

    The Galapagos Microplate (GM) lies on the western Gaplapagos Spreading Center (GSC), representing one of the classic Ridge-Ridge-Ridge (R-R-R) plate boundaries of the Nazca, Cocos, and Pacific plates. The presence of the 'black smoke' and hydrothermal vent community were firstly confirmed on the GSC. Lots of hydrothermal fields were discovered on the center and eastern GSC, while the western GSC has not been well investigated. During 17th Oct. to 9th Nov. 2009, the 3rd leg of Chinese DY115-21 cruise with R/V Dayangyihao has been launched along 2°N-5°S near equatorial East Pacific Rise (EPR). Two new hydrothermal fields were confirmed. One is named 'Precious Stone Mountain', which is the first hydrothermal field on the GM. The other is found at 101.47°W, 0.84°S EPR. The 'Precious Stone Mountain' hydrothermal field (at 101.49°W, 1.22°N) is located at an off-axial seamount on the southern GM boundary, with a depth from 1,450 to 1,700m. Hydrothermal fluids emitting from the fissures and hydrothermal fauna were captured by deep-tow video. Few mineral clasts of pyrite and chalcopyrite were separated from one sediment sample, but no sulfide chimney was found yet. Hydrothermal fauna such as alive mussels, crabs, shrimps, tubeworms, giant clams, as well as rock samples were collected by TV-Grab. The study of the seafloor classification with Simrad EM120 multi-beam echosounder has been conducted on the 'Precious Stone Mountain' hydrothermal field. The result indicates that seafloor materials around the hydrothermal field can be characterized into three types, such as the fresh lava, hydrothermal sediment, and altered rock.

  1. 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

  2. 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. PMID:27389518

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

    NASA Astrophysics Data System (ADS)

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

    2016-08-01

    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.

  4. 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). PMID:25103033

  5. 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. PMID:26738495

  6. 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)

  7. 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. PMID:25289516

  8. 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.

  9. Vigorous venting and biology at Pito Seamount, Easter microplate

    NASA Astrophysics Data System (ADS)

    Naar, D. F.; Hekinian, R.; Segonzac, M.; Francheteau, J.; Armijo, R.; Cogne, J.-P.; Constantin, M.; Girardeau, J.; Hey, R. N.; Searle, R. C.

    A Nautile submersible investigation of Pito Seamount documents vigorous hydrothermal venting at 23°19.65'S, 111°38.41'W and at a depth of 2270 m. The data indicate the volcano is young and recently active, as predicted from analyses of SeaMARC II side-scan and swath bathymetry, and geophysical data. Pito Seamount lies near Pito Deep (5980 m), which marks the tip of the northwestward propagating East rift of the Easter microplate. Bathymetry surrounding Pito Seamount consists of a series of ridges and valleys with relief up to ˜4 km. The 4-km submersible-transect to the summit of Pito Seamount crossed areas of very glassy basalt with little or no sediment cover, suggesting the lava flows are very young. Most of the lava samples from Pito Seamount are depleted normal MORB (mid-ocean ridge basalt). Lava samples associated with active and dead hydrothermal vents consist of phyric and aphyric transitional and enriched MORB. Sulfides consist primarily of sphalerite and pyrite, with traces of chalcopyrite. The active hydrothermal chimney on Pito Seamount has a small, undiversified biological community similar to northern East Pacific Rise vent sites (alvinellid worms, bythograeid crabs and bythitid fishes) and western Pacific back-arc basin sites (alvinocaridid shrimps). No vestimentiferan worms were observed. Previous geophysical data, and new geochemical data and visual observations, suggest that the vigorous black smoker is a result of deep, extensive crosscutting faults formed by extensive tectonic thinning of Pito Deep, and a very robust magmatic supply being supplied from upwelling asthenosphere. Although no biological or vent fluid samples were obtained, geological and biological observations, such as the large number of inactive chimneys, old hydrothermal deposits, and starfish, as well as the occurrence of dead mollusks (gastropod and mussels), suggest a recent waning of hydrothermal activity near the summit. The speculative interpretation that Pito Seamount

  10. 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

  11. 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.

  12. 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

  13. Measurement of filter paper activities of cellulase with microplate-based assay

    PubMed Central

    Yu, Xiaoxiao; Liu, Yan; Cui, Yuxiao; Cheng, Qiyue; Zhang, Zaixiao; Lu, Jia Hui; Meng, Qingfan; Teng, Lirong; Ren, Xiaodong

    2015-01-01

    It is always a challenge to determine the total cellulase activity efficiently without reducing accuracy. The most common total cellulase activity assay is the filter paper assay (FPA) established by the International Union of Pure and Applied Chemistry (IUPAC). A new procedure to measure the FPA with microplate-based assay was studied in this work, which followed the main idea of IUPAC to dilute cellulase preparation to get fixed glucose release. FPAs of six cellulase preparations were determined with the microplate-based assay. It is shown that FPAs of cellulase Youtell, RCconc, R-10, Lerkam, Yishui and Sinopharm were 67.9, 46.0, 46.1, 27.4, 7.6 and 8.0 IU/ml respectively. There was no significant difference at the 95% confidence level between the FPA determined with IUPAC and the microplate-based assay. It could be concluded that the FPA could be determined by the microplate-based assay with the same accuracy and much more efficiency compared with that by IUPAC. PMID:26858572

  14. Microplate based optical biosensor for L-Dopa using tyrosinase from Amorphophallus campanulatus.

    PubMed

    Saini, Amardeep Singh; Kumar, Jitendra; Melo, Jose Savio

    2014-11-01

    Developing a biosensor which is capable of simultaneously monitoring l-Dopa levels in multiple samples besides requiring small reaction volume is of great value. The present study describes the detection of l-Dopa using tyrosinase enzyme extracted from Amorphophallus campanulatus and immobilized on the surface of the microplate wells. Among the different approaches used for immobilizing tyrosinase onto the microplate wells, glutaraldehyde treatment was found to be most effective. Besides enzyme activity, ESEM-EDS (environmental scanning electron microscope-energy dispersive system) and Atomic Force Microscopy (AFM) were also carried out to confirm the immobilization of tyrosinase enzyme onto the microplate well surface. This immobilized biocomponent was then integrated with an optical transducer for l-Dopa detection and it showed good reproducibility. The sensing property of the system was studied by measuring the initial rate of dopachrome formation at 475 nm. The calibration plot gave a linear range of detection from 10-1000 μM and the detection limit was calculated to be 3 μM. The immobilized biocomponent was stable for 41 days and was reused up to nine times. Spiked samples (blood plasma) were also analyzed using this biocomponent. This microplate based biosensor thus provides a convenient system for detection of multiple samples in a single run. PMID:25300217

  15. "Liquid Knife" to Fabricate Patterning Single-Crystalline Perovskite Microplates toward High-Performance Laser Arrays.

    PubMed

    Feng, Jiangang; Yan, Xiaoxu; Zhang, Yifan; Wang, Xuedong; Wu, Yuchen; Su, Bin; Fu, Hongbin; Jiang, Lei

    2016-05-01

    A facile and effective "liquid knife" is created by controlling the dewetting process of the liquid precursor, yielding patterning single-crystalline perovskite microplates with uniform size, precise positioning, high quality, and low lasing thresholds. The sizes and location of single-crystalline perovskite are controllable, leading to mode-tunable lasing emission and patterned lasers. PMID:27000628

  16. Measurement of filter paper activities of cellulase with microplate-based assay.

    PubMed

    Yu, Xiaoxiao; Liu, Yan; Cui, Yuxiao; Cheng, Qiyue; Zhang, Zaixiao; Lu, Jia Hui; Meng, Qingfan; Teng, Lirong; Ren, Xiaodong

    2016-01-01

    It is always a challenge to determine the total cellulase activity efficiently without reducing accuracy. The most common total cellulase activity assay is the filter paper assay (FPA) established by the International Union of Pure and Applied Chemistry (IUPAC). A new procedure to measure the FPA with microplate-based assay was studied in this work, which followed the main idea of IUPAC to dilute cellulase preparation to get fixed glucose release. FPAs of six cellulase preparations were determined with the microplate-based assay. It is shown that FPAs of cellulase Youtell, RCconc, R-10, Lerkam, Yishui and Sinopharm were 67.9, 46.0, 46.1, 27.4, 7.6 and 8.0 IU/ml respectively. There was no significant difference at the 95% confidence level between the FPA determined with IUPAC and the microplate-based assay. It could be concluded that the FPA could be determined by the microplate-based assay with the same accuracy and much more efficiency compared with that by IUPAC. PMID:26858572

  17. Coupling the Torpedo Microplate-Receptor Binding Assay with Mass Spectrometry to Detect Cyclic Imine Neurotoxins

    PubMed Central

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

    2014-01-01

    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. PMID:23131021

  18. 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

    2012-12-01

    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. PMID:23131021

  19. Customizable PCR-microplate array for differential identification of multiple pathogens

    PubMed Central

    Woubit, Abdela; Yehualaeshet, Teshome; Roberts, Sherrelle; Graham, Martha; Kim, Moonil; Samuel, Temesgen

    2014-01-01

    Customizable PCR-microplate arrays were developed for the rapid identification of Francisella tularensis subsp. tularensis, Salmonella Typhi, Shigella dysenteriae, Yersinia pestis, Vibrio cholerae Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Saintpaul, Francisella tularensis subsp. novicida, Vibrio parahaemolyticus, and Yersinia pseudotuberculosis. Previously, we identified highly specific primers targeting each of the pathogens above. Here, we report the development of customizable PCR-microplate arrays for simultaneous identification of the pathogens using the primers. A mixed aliquot of genomic DNA from 38 different strains was used to validate three PCR-microplate array formats. Identical PCR conditions were used to run all the samples on the three formats. Results show specific amplifications on all the three custom plates. In a preliminary test to evaluate the sensitivity of these assays in laboratory-inoculated samples, detection limits as low as 9 cfu/g/ml S. Typhimurium were obtained from beef hot dog, and 78 cfu/ml from milk. Such microplate arrays could serve as valuable tools for initial identification or secondary confirmation of these pathogens. PMID:24215700

  20. High-Resolution Vertical Observation of Intracellular Structure Using Magnetically Responsive Microplates.

    PubMed

    Teshima, Tetsuhiko; Onoe, Hiroaki; Tottori, Soichiro; Aonuma, Hiroka; Mizutani, Takeomi; Kamiya, Koki; Ishihara, Hirotaka; Kanuka, Hirotaka; Takeuchi, Shoji

    2016-07-01

    A vertical confocal observation system capable of high-resolution observation of intracellular structure is demonstrated. The system consists of magnet-active microplates to rotate, incline, and translate single adherent cells in the applied magnetic field. Appended to conventional confocal microscopes, this system enables high-resolution cross-sectional imaging with single-molecule sensitivity in single scanning. PMID:27185344

  1. Simultaneous determination of a novel KDR kinase inhibitor and its N-oxide metabolite in human plasma using 96-well solid-phase extraction and liquid chromatography/tandem mass spectrometry.

    PubMed

    Xu, Yang; Du, Lihong; Soli, Eric D; Braun, Matthew P; Dean, Dennis C; Musson, Donald G

    2005-03-25

    To support pharmacokinetic studies, a selective and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous determination of a novel KDR kinase inhibitor (1) and its active metabolite (2) in human plasma. The method is fully automated using a Packard MultiPROBE II system and a TomTec Quadra 96 liquid handling workstation to perform sample preparation and solid-phase extraction (SPE). Following the extraction on a mixed-mode SPE using Oasis MCX 96-well plate, the analytes were separated on a Aquasil C18 column (50 mm x 2.1 mm, i.d., 3 microm) with a mobile phase consisting of acetonitrile/ammonium acetate buffer (5 mM, pH 5.0) (60/40, v/v). The run time for each injection was 4.5 min with the retention times of approximately 2.0 and 2.7 min for 1 and 2 respectively, at a flow rate of 0.25 mL/min. A tandem mass spectrometric detection was conducted using multiple reaction monitoring (MRM) under the positive ion mode with a turbo ion-spray interface. The linear ranges of the calibration curves were 0.05-400 ng/mL for 1 and 0.1-400 ng/mL for 2 on a PE Sciex API 4000 LC-MS/MS system. The lower limits of quantitation (LLOQ) of the assay were 0.05 and 0.1 ng/mL for 1 and 2 respectively, when 0.4 mL of plasma was processed. Intra-day assay precision (using five standard curves prepared by spiking compounds to five lots of plasma) was less than 4.9% for 1 and less than 9.6% for 2 on each concentration. Assay accuracy was found to be 95.1-104.6% of nominal for 1 standards and 93.5-105.6% for 2 standards. QC samples were stable when kept at room temperature for 4 h, at -70 degrees C for 10 days, and after three freeze-thaw cycles. The extraction recoveries were 80%, 83% and 84% for 1 and 2 and I.S. respectively, and no significant matrix effects were observed. The method was successfully applied to plasma samples from clinical studies after oral administration of compound 1. PMID:15686997

  2. 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.

  3. The Westcoast Reader. A Newspaper for New Adult Readers.

    ERIC Educational Resources Information Center

    Acosta, Joan, Ed.

    This document consists of informative material on and a copy of a newspaper for adults who are learning to read. The newspaper is designed to help new readers develop reading skills, while providing interesting and relevant information with an adult focus. Local, national, and international news stories as well as information about health, safety,…

  4. Turkish Basic Course. Graded Reader.

    ERIC Educational Resources Information Center

    Agrali, Selman N., Comp.; And Others

    The present Reader is the third and final volume in the Foreign Service Institute's "Turkish Basic Course." (See ED 013 451 and ED 024 050 for Units 1-30 and Units 31-50.) Reading selections are arranged in approximate ascending order of difficulty with some grouping of selections in subject matter categories. The selections, which have not been…

  5. 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…

  6. 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…

  7. 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.

  8. 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;…

  9. A Matchmaking Tool for Readers.

    ERIC Educational Resources Information Center

    Belcastro, Patricia

    1995-01-01

    Discusses readers' advisory services in public libraries and describes a method to convert information on specific books and genres to a computer database. Topics include utilizing staff knowledge, fields included for each book, the development of descriptors, the editing process, and writing summaries and critiques of the books. (LRW)

  10. 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…

  11. Native American Adult Reader III.

    ERIC Educational Resources Information Center

    King, Lovern Root, Ed.

    This reader, one of three designed to provide adults in basic education/GED programs with meaningful material based on Native American cultures, includes selections appropriate for advanced reading ability (grade 7 and above). The twelve readings focus on culture, history, and contemporary concerns of Native Americans. Each selection includes a…

  12. Native American Adult Reader II.

    ERIC Educational Resources Information Center

    King, Lovern Root, Ed.

    This reader, one of three designed to provide adults in basic education/GED programs with meaningful materials based on Native American cultures, includes selections appropriate for intermediate reading ability (grades 4-6). The twelve readings focus on culture, history, and contemporary concerns of Native Americans. Each selection includes a…

  13. Raising Rigor for Struggling Readers

    ERIC Educational Resources Information Center

    Taylor, Rosemarye T.; Watson, Rebecca

    2013-01-01

    Increasing the rigor of learning tasks to align with the expectations in the Common Core State Standards presents challenges for many educators, particularly for middle and high school teachers of nonproficient readers. In this article, the authors describe a number of these challenges and identify four steps that together, create a systematic…

  14. Predictable Books: Captivating Young Readers.

    ERIC Educational Resources Information Center

    Luckner, John

    1990-01-01

    Because prediction plays such a vital role in reading comprehension, predictable books are essential in the teaching of beginning readers. Prediction involves a three-step cycle: sampling, predicting, and confirming. Steps in using predictable books with hearing-impaired students are outlined, and a list of predictable and repetitive books is…

  15. 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…

  16. Reading Comprehension for Older Readers

    ERIC Educational Resources Information Center

    Vaughn, Sharon; Edmonds, Meaghan

    2006-01-01

    This article provides an overview of a multicomponent comprehension strategy and graphic organizers designed for older readers to gain meaning from text. Practices designed to capitalize on the best research-based elements associated with improved outcomes in reading comprehension, particularly for expository texts, are described. The graphic…

  17. 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…

  18. Fast and Accurate Microplate Method (Biolog MT2) for Detection of Fusarium Fungicides Resistance/Sensitivity

    PubMed Central

    Frąc, Magdalena; Gryta, Agata; Oszust, Karolina; Kotowicz, Natalia

    2016-01-01

    The need for finding fungicides against Fusarium is a key step in the chemical plant protection and using appropriate chemical agents. Existing, conventional methods of evaluation of Fusarium isolates resistance to fungicides are costly, time-consuming and potentially environmentally harmful due to usage of high amounts of potentially toxic chemicals. Therefore, the development of fast, accurate and effective detection methods for Fusarium resistance to fungicides is urgently required. MT2 microplates (BiologTM) method is traditionally used for bacteria identification and the evaluation of their ability to utilize different carbon substrates. However, to the best of our knowledge, there is no reports concerning the use of this technical tool to determine fungicides resistance of the Fusarium isolates. For this reason, the objectives of this study are to develop a fast method for Fusarium resistance to fungicides detection and to validate the effectiveness approach between both traditional hole-plate and MT2 microplates assays. In presented study MT2 microplate-based assay was evaluated for potential use as an alternative resistance detection method. This was carried out using three commercially available fungicides, containing following active substances: triazoles (tebuconazole), benzimidazoles (carbendazim) and strobilurins (azoxystrobin), in six concentrations (0, 0.0005, 0.005, 0.05, 0.1, 0.2%), for nine selected Fusarium isolates. In this study, the particular concentrations of each fungicides was loaded into MT2 microplate wells. The wells were inoculated with the Fusarium mycelium suspended in PM4-IF inoculating fluid. Before inoculation the suspension was standardized for each isolates into 75% of transmittance. Traditional hole-plate method was used as a control assay. The fungicides concentrations in control method were the following: 0, 0.0005, 0.005, 0.05, 0.5, 1, 2, 5, 10, 25, and 50%. Strong relationships between MT2 microplate and traditional hole

  19. Wandering terranes in southern Alaska: The Aleutia Microplate and implications for the Bering Sea

    NASA Astrophysics Data System (ADS)

    Marlow, Michael S.; Cooper, Alan K.

    1983-04-01

    Paleomagnetic and geological data suggest that much of southern Alaska is a collage of tectonostratigraphic terranes which originated in Mesozoic time at paleolatitudes far south of their present position. The time of `docking' of the terranes against cratonic Alaska is critical to defining their amalgamated size and extent during their northward motion as well as their role in the evolution of the Bering Sea. One of the largest of the tectonostratigraphic terranes, the Peninsular terrane of south central and southwestern Alaska, extends offshore along the outer Bering Sea continental margin (Beringia). Paleomagnetic data suggest that this terrane has moved northward through all of Cenozoic time, but geologic data imply that the terrane had accreted to Alaska by the end of the Mesozoic. In early Cenozoic time the eastern part of the Aleutian arc appears to have been superimposed on the Peninsular terrane, and postulated northward Cenozoic motion of the terrane would therefore have required northward motion of the arc. Two accretion models, based on docking times for terranes in Alaska, are proposed, and they illustrate that large areas of the abyssal Bering Sea, the Alaska Peninsula, the Aleutian arc, and the Beringian continental margin may be part of a superterrane or microplate called Aleutia (microplate as defined by Beck et al. (1980), i.e., a microplate is a displaced segment of lithosphere that has crustal roots, whereas a superterrane is an amalgamation of terranes which may or may not be rootless). Model A implies that the Aleutian arc developed in situ on the southern edge of Aleutia after the microplate had docked. In model B, the final docking time of the Peninsular terrane is late Cenozoic, which implies that the Aleutia microplate encompasses a mammoth area that includes parts of southern Alaska, the Alaska Peninsula, the southern Beringian margin, the abyssal Bering Sea (Kula plate), and the Aleutian arc. If model A is correct, the docking time of

  20. Fast and Accurate Microplate Method (Biolog MT2) for Detection of Fusarium Fungicides Resistance/Sensitivity.

    PubMed

    Frąc, Magdalena; Gryta, Agata; Oszust, Karolina; Kotowicz, Natalia

    2016-01-01

    The need for finding fungicides against Fusarium is a key step in the chemical plant protection and using appropriate chemical agents. Existing, conventional methods of evaluation of Fusarium isolates resistance to fungicides are costly, time-consuming and potentially environmentally harmful due to usage of high amounts of potentially toxic chemicals. Therefore, the development of fast, accurate and effective detection methods for Fusarium resistance to fungicides is urgently required. MT2 microplates (Biolog(TM)) method is traditionally used for bacteria identification and the evaluation of their ability to utilize different carbon substrates. However, to the best of our knowledge, there is no reports concerning the use of this technical tool to determine fungicides resistance of the Fusarium isolates. For this reason, the objectives of this study are to develop a fast method for Fusarium resistance to fungicides detection and to validate the effectiveness approach between both traditional hole-plate and MT2 microplates assays. In presented study MT2 microplate-based assay was evaluated for potential use as an alternative resistance detection method. This was carried out using three commercially available fungicides, containing following active substances: triazoles (tebuconazole), benzimidazoles (carbendazim) and strobilurins (azoxystrobin), in six concentrations (0, 0.0005, 0.005, 0.05, 0.1, 0.2%), for nine selected Fusarium isolates. In this study, the particular concentrations of each fungicides was loaded into MT2 microplate wells. The wells were inoculated with the Fusarium mycelium suspended in PM4-IF inoculating fluid. Before inoculation the suspension was standardized for each isolates into 75% of transmittance. Traditional hole-plate method was used as a control assay. The fungicides concentrations in control method were the following: 0, 0.0005, 0.005, 0.05, 0.5, 1, 2, 5, 10, 25, and 50%. Strong relationships between MT2 microplate and traditional hole

  1. 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

  2. Into Focus: Understanding and Creating Middle School Readers.

    ERIC Educational Resources Information Center

    Beers, Kylene, Ed.; Samuels, Barbara G., Ed.

    Addressing the needs of English language learners, remedial readers, avid readers, gifted readers, dormant readers, reluctant readers, and readers in content areas, this book's goal is to help middle school teachers connect their students to reading. The book offers multiple lists of trade books that different types of middle school readers enjoy;…

  3. Analysis of Aerobic Respiration in Intact Skeletal Muscle Tissue by Microplate-Based Respirometry.

    PubMed

    Shintaku, Jonathan; Guttridge, Denis C

    2016-01-01

    Mitochondrial function is a key component of skeletal muscle health, and its dysfunction has been associated with a wide variety of diseases. Microplate-based respirometry measures aerobic respiration of live cells through extracellular changes in oxygen concentration. Here, we describe a methodology to measure aerobic respiration of intact murine skeletal muscle tissue. The tissues are not cultured, permeabilized, or enzymatically dissociated to single fibers, so there is minimal experimental manipulation affecting the samples prior to acquiring measurements. PMID:27492183

  4. Carbon-coated lithium titanium phosphate nanoporous microplates with superior electrochemical performance

    NASA Astrophysics Data System (ADS)

    Huang, Zhifeng; Liu, Li; Zhou, Qian; Tan, Jinli; Yan, Zichao; Xia, Dongdong; Shu, Hongbo; Yang, Xiukang; Wang, Xianyou

    2015-10-01

    In this study, we report a facile method to synthesize carbon-coated LiTi2(PO4)3 nanoporous microplates (LTP/C MPs) using ethylenediamine as the chelating agent and carbon source. The as-prepared LTP/C MPs with thickness of 0.4 μm consist of interconnected nanosized particles embedded in nano-thickness carbon layer and well-dispersed nanopores. The carbon layer significantly improves the electrochemical performance of LiTi2(PO4)3 microplates. LTP/C MPs deliver a reversible capacity of 121 mAh g-1 at 0.2C (1C = 138 mAh g-1) and show a remarkable capacity retention of 94.2% over 100 cycles when matched with Li metal counter electrode. It also presents excellent electrochemical properties as anode material for aqueous rechargeable lithium batteries (ARLBs). LTP/C MPs//LiMn2O4 ARLB shows a high discharge capacity of 76 mAh g-1 at 20 mA g-1 and superior rate capability. The results suggest a practical stratagem to develop a novel composite in which the carbon is coated with LiTi2(PO4)3 nanoporous microplates, which can become one of the promising electrode materials for both non-aqueous and aqueous lithium ion batteries.

  5. Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements

    PubMed Central

    Boutagy, Nabil E.; Rogers, George W.; Pyne, Emily S.; Ali, Mostafa M.; Hulver, Matthew W.; Frisard, Madlyn I.

    2015-01-01

    Skeletal muscle mitochondria play a specific role in many disease pathologies. As such, the measurement of oxygen consumption as an indicator of mitochondrial function in this tissue has become more prevalent. Although many technologies and assays exist that measure mitochondrial respiratory pathways in a variety of cells, tissue and species, there is currently a void in the literature in regards to the compilation of these assays using isolated mitochondria from mouse skeletal muscle for use in microplate based technologies. Importantly, the use of microplate based respirometric assays is growing among mitochondrial biologists as it allows for high throughput measurements using minimal quantities of isolated mitochondria. Therefore, a collection of microplate based respirometric assays were developed that are able to assess mechanistic changes/adaptations in oxygen consumption in a commonly used animal model. The methods presented herein provide step-by-step instructions to perform these assays with an optimal amount of mitochondrial protein and reagents, and high precision as evidenced by the minimal variance across the dynamic range of each assay. PMID:26555567

  6. Platelet aggregation inhibitors from Philippine marine invertebrate samples screened in a new microplate assay.

    PubMed

    Pimentel, Sheila Marie V; Bojo, Zenaida P; Roberto, Amy V D; Lazaro, Jose Enrico H; Mangalindan, Gina C; Florentino, Leila M; Lim-Navarro, Pilar; Tasdemir, Deniz; Ireland, Chris M; Concepcion, Gisela P

    2003-01-01

    A new microplate assay for Ca(2+)-induced platelet aggregation as detected by Giemsa dye was used to screen marine invertebrate samples from the Philippines for inhibitors of human platelet aggregation. Out of 261 crude methanol extracts of marine sponges and tunicates, 25 inhibited aggregation at 2 mg/ml. Inhibition of agonist-induced aggregation in an aggregometer was used to confirm results of the microplate assay and to determine the specific mode of inhibition of 2 samples. The marine sponge Xestospongia sp. yielded a xestospongin/araguspongine-type molecule that inhibited collagen-induced aggregation by 87% at 2 micro g/ml, and epinephrine-induced aggregation by 78% at 20 micro g/ml, while the marine sponge Aplysina sp. yielded 5,6-dibromotryptamine, which inhibited epinephrine-induced aggregation by 51% at 20 micro g/ml. In this study we have found that the microplate assay is a simple, inexpensive, yet useful preliminary tool to qualitatively screen a large number of marine samples for antiplatelet aggregation activity. PMID:14719168

  7. 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

  8. Influence of acoustic streaming on ultrasonic particle manipulation in a 100-well ring-transducer microplate

    NASA Astrophysics Data System (ADS)

    Ohlin, Mathias; Christakou, Athanasia E.; Frisk, Thomas; Önfelt, Björn; Wiklund, Martin

    2013-03-01

    We characterize and quantify the performance of ultrasonic particle aggregation and positioning in a 100-well microplate. We analyze the result when operating a planar ultrasonic ring transducer at different single actuation frequencies in the range 2.20-2.40 MHz, and compare with the result obtained from different schemes of frequency-modulated actuation. Compared to our previously used wedge transducer design, the ring transducer has a larger contact area facing the microplate, resulting in lower temperature increase for a given actuation voltage. Furthermore, we analyze the dynamics of acoustic streaming occurring simultaneously with the particle trapping in the wells of the microplate, and we define an adaptive ultrasonic actuation scheme for optimizing both efficiency and robustness of the method. The device is designed as a tool for ultrasound-mediated cell aggregation and positioning. This is a method for high-resolution optical characterization of time-dependent cellular processes at the level of single cells. In this paper, we demonstrate how to operate our device in order to optimize the scanning time of 3D confocal microscopy with the aim to perform high-resolution time-lapse imaging of cells or cell-cell interactions in a highly parallel manner.

  9. Graded Readers: Validating Reading Levels across Publishers

    ERIC Educational Resources Information Center

    Rodrigo, Victoria

    2016-01-01

    Graded readers can be an optimal resource to help language students improve and personalize their learning experience. An extensive reading library with graded readers and well-defined levels of reading difficulty increases language students' chances of having a successful reading experience and become independent readers. However, when it comes…

  10. Enhancing Readers' Analysis-by-Synthesis Abilities.

    ERIC Educational Resources Information Center

    McIntosh, Margaret E.

    A variety of techniques for improving readers' analysis-by-synthesis abilities (rapid, efficient reading typical of highly skilled readers) are presented in this paper. The techniques discussed in the first part emphasize improving reading comprehension and include the following: (1) modifications of the cloze procedure (encouraging readers to use…

  11. 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.…

  12. Precision Reading: New Hope for Struggling Readers

    ERIC Educational Resources Information Center

    Freeze, Rick

    2006-01-01

    A rationale for a novel approach to remedial reading for struggling readers, including students labeled with intellectual disabilities, learning disabilities, attention deficit and behavior disorders, below average language development, and those described as reluctant or resistant readers is presented. It is suggested that struggling readers find…

  13. 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…

  14. Extending the E-Z Reader Model of Eye Movement Control to Chinese Readers

    ERIC Educational Resources Information Center

    Rayner, Keith; Li, Xingshan; Pollatsek, Alexander

    2007-01-01

    Chinese readers' eye movements were simulated in the context of the E-Z Reader model, which was developed to account for the eye movements of readers of English. Despite obvious differences between English and Chinese, the model did a fairly good job of simulating the eye movements of Chinese readers. The successful simulation suggests that the…

  15. 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

  16. Enzyme activity deviates due to spatial and temporal temperature profiles in commercial microtiter plate readers.

    PubMed

    Grosch, Jan-Hendrik; Sieben, Michaela; Lattermann, Clemens; Kauffmann, Kira; Büchs, Jochen; Spieß, Antje C

    2016-03-01

    Microtiter plates (MTP) and automatized techniques are increasingly applied in the field of biotechnology. However, the susceptibility of MTPs to edge effects such as thermal gradients can lead to high variation of measured enzyme activities. In an effort to enhance experimental reliability, to quantify, and to minimize instrument-caused deviations in enzyme kinetics between two MTP-readers, we comprehensively quantified temperature distribution in 96-well MTPs. We demonstrated the robust application of the absorbance dye cresol red as easily applicable temperature indicator in cuvettes and MTPs and determined its accuracy to ±0.16°C. We then quantified temperature distributions in 96-well MTPs revealing temperature deviations over single MTP of up to 2.2°C and different patterns in two commercial devices (BioTek Synergy 4 and Synergy Mx). The obtained liquid temperature was shown to be substantially controlled by evaporation. The temperature-induced enzyme activity variation within MTPs amounted to about 20 %. Activity deviations between MTPs and to those in cuvettes were determined to 40 % due to deviations from the set temperature in MTPs. In conclusion, we propose a better control of experimental conditions in MTPs or alternative experimental systems for reliable determination of kinetic parameters for bioprocess development. PMID:26709721

  17. 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.

  18. 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

  19. Variation in CR imaging plate readers.

    PubMed

    Fauber, Terri L; Legg, Jeffrey S; Quinn, Megan

    2002-01-01

    This study investigated computed radiography (CR) image processing to determine whether variation exists within and among CR imaging plate readers. Photostimulable imaging plates were exposed using a phantom test tool and processed in 4 CR readers located in diverse settings in an urban academic medical center. Research results indicate daily variation of S-numbers within individual CR readers did not exceed tolerance limits, although over the 3-week study period, evidence of S-number variation within individual CR readers was mixed. In addition, S-number variability among multiple CR readers was found to be statistically significant. Although the cause of the variability remains unknown, evidence of variability among multiple CR readers indicates a need for systematic quality control. PMID:12362532

  20. 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.

  1. The August 2002 earthquake sequence in north Afar: Insights into the neotectonics of the Danakil microplate

    NASA Astrophysics Data System (ADS)

    Ayele, Atalay; Stuart, Graham; Bastow, Ian; Keir, Derek

    2007-06-01

    In August 2002, there was high seismic activity in Afar concentrated at the plateau margin of the northern Ethiopian rift east of Mekele, near the western part of the Danakil microplate. The spatial and temporal distributions of this seismic activity over four weeks indicate the NNW propagation of the Gulf of Aden rift across the Afar Depression towards the western Ethiopian plateau. Fault plane solutions for six larger earthquakes from the August 2002 sequence are estimated from moment tensor inversion of local broadband waveform data. The results show only normal faulting on NNW trending and NE dipping faults, which agree with tectonics of the area and distribution of aftershocks. No strike-slip component is observed in any of our fault plane solutions or those of other workers including Harvard CMT solutions in the region. Such motion would be indicative of oblique-slip deformation between the Nubian plate and the Danakil microplate consistent with counter-clockwise rotation of the microplate. Hypocentral depths of well-constrained events are 5-7 km, which is the approximate elastic plate thickness in the Main Ethiopian rift, possibly indicating the depth to the brittle-ductile transition zone in this part of the Afar Depression. The shallowness of the depth estimates agree with the macroseismic reports available from a wide area in northern Ethiopia. Potential future shallow crustal deformation may cause significant loss of human life and damage to property in the densely populated highland region around Mekele unless measures are taken in improving building standards. The b-value for this sequence is estimated to be 0.66 using a least squares fit, while it is 0.67 ± 0.16 from a maximum-likelihood approach. This estimated b-value is low or the frequency of occurrence of relatively larger magnitude events is high indicating that it is a highly stressed region as evidenced by the recent increase of the seismicity in the area.

  2. The Geometry of the Subducting Slabs Beneath the PRVI Microplate Based on 3D Tomography

    NASA Astrophysics Data System (ADS)

    Xu, X.; Keller, G. R.; Holland, A. A.; Keranen, K. M.; Li, H.

    2011-12-01

    The Puerto Rico and the Virgin Islands (PRVI) microplate is located between two subduction zones, with the Puerto Rico trench to the north and the Muertos trough to the south. The Puerto Rico trench is caused by southward-directed subduction of the North American Plate, and the Muertos trough is the northern boundary of the Caribbean Plate. There is no active volcanism on Puerto Rico; however, earthquake depths and seismic tomography imply that the slab of Caribbean plate continues northward beneath Puerto Rico. Puerto Rico overlies these two slabs with extension to both the west (Mona Passage) and southeast (Anacapa Passage). The cause of the extension is unknown, but GPS measurements show that Puerto Rico is experiencing anti-clockwise rotation, and the extension associated with the Anacapa Passage may be produced by this rotation. To the west, it is debated whether the Mona Passage is a boundary between two micro-plates or simple a local rift basin. To address the sources of the extension and the cause of the rotation, we are investigating if the deep structures can be the dynamic source for the observed kinematic movements. We collected data on earthquakes occurring between 2009-2011 in the PRVI region and relocated them using the SEISAN code provided by the Institute of Solid Earth Physics, University of Bergen. The FMTOMO code from Australian National University was used for 3D tomography from P and S wave arrival times. By comparing the relocated epicenters and the 3D tomography results, the subducting slabs were identified. When integrated with the results of previous studies, the geometric model of the slabs is a critical key to understanding the evolution of the PRVI microplate in the past and the future.

  3. 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.

  4. Self-assembly of gold nanorods into vertically aligned, rectangular microplates with a supercrystalline structure

    NASA Astrophysics Data System (ADS)

    Xiao, Junyan; Li, Zhe; Ye, Xiaozhou; Ma, Yurong; Qi, Limin

    2013-12-01

    Vertically aligned, supercrystalline microplates with a well-defined rectangular shape were fabricated in a large area through self-assembly of gold nanorods by a novel bulk solution evaporation method. This evaporative self-assembly strategy involving continuous movement of the contact line can prevent the coffee-ring effect, thus allowing uniform deposition of discrete GNR superstructures over a large area and favoring the formation of GNR supercrystals with geometrically symmetric shapes. A mechanism based on the continuing nucleation and growth of smectic GNR superstructures accompanying the movement of the contact line was put forward for the formation of the unique GNR supercrystal arrays. Based on this mechanism, a micropatterned substrate was designed to control the nucleation location and growth direction, leading to the spontaneous self-assembly of nearly parallel arrays of vertically aligned, supercrystalline microplates of GNRs. The obtained rectangular-plate-shaped GNR supercrystals exhibited interesting anisotropic optical reflection properties, which were revealed by polarized light microscopy.Vertically aligned, supercrystalline microplates with a well-defined rectangular shape were fabricated in a large area through self-assembly of gold nanorods by a novel bulk solution evaporation method. This evaporative self-assembly strategy involving continuous movement of the contact line can prevent the coffee-ring effect, thus allowing uniform deposition of discrete GNR superstructures over a large area and favoring the formation of GNR supercrystals with geometrically symmetric shapes. A mechanism based on the continuing nucleation and growth of smectic GNR superstructures accompanying the movement of the contact line was put forward for the formation of the unique GNR supercrystal arrays. Based on this mechanism, a micropatterned substrate was designed to control the nucleation location and growth direction, leading to the spontaneous self-assembly of

  5. [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. PMID:6913282

  6. 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…

  7. 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...

  8. Nanoshell-Enhanced Raman Spectroscopy on a Microplate for Staphylococcal Enterotoxin B Sensing.

    PubMed

    Wang, Wenbin; Wang, Weiwei; Liu, Liqiang; Xu, Liguang; Kuang, Hua; Zhu, Jianping; Xu, Chuanlai

    2016-06-22

    A sensitive surface-enhanced Raman scattering (SERS) immunosensor based on the Au nanoparticle (Au NP) shell structure was developed to detect staphylococcal enterotoxin B (SEB) on a microplate. Au NPs modified with 4-nitrothiophenol (4-NTP) and coated with Ag shell of controlled thickness at 6.6 nm exhibited excellent SERS intensity and were used as signal reporters in the detection of SEB. The engaged 4-NTP allowed the significant electromagnetic enhancement between Au NPs and the Ag shell and prevented the dissociation of the Raman reporter. More importantly, 4-NTP-differentiated SERS signals between the sample and microplate. The SERS-based immunosensor had a limit of detection of 1.3 pg/mL SEB. Analysis of SEB-spiked milk samples revealed that the developed method had high accuracy. Therefore, the SERS-encoded Au@Ag core-shell structure-based immunosensor is promising for the detection of biotoxins, pathogens, and environmental pollutants. PMID:27193082

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

    PubMed

    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

  10. 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

  11. 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.

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

    PubMed

    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

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

    PubMed

    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

  14. An Analytical Model for Squeeze-Film Damping of Perforated Torsional Microplates Resonators

    PubMed Central

    Li, Pu; Fang, Yuming

    2015-01-01

    Squeeze-film damping plays a significant role in the performance of micro-resonators because it determines their quality factors. Perforations in microstructures are often used to control the squeeze-film damping in micro-resonators. To model the perforation effects on the squeeze-film damping, many analytical models have been proposed, however, most of the previous models have been concerned with the squeeze-film damping due to the normal motion between the perforated vibrating plate and a fixed substrate, while there is a lack of works that model the squeeze-film damping of perforated torsion microplates, which are also widely used in MEMS devices. This paper presents an analytical model for the squeeze-film damping of perforated torsion microplates. The derivation in this paper is based on a modified Reynolds equation that includes compressibility and rarefaction effects. The pressure distribution under the vibrating plate is obtained using the double sine series. Closed-form expressions for the stiffness and the damping coefficients of the squeeze-film are derived. The accuracy of the model is verified by comparing its results with the finite element method (FEM) results and the experimental results available in the literature. The regime of validity and limitations of the present model are assessed. PMID:25815453

  15. Biocompatibility of microplates for culturing epithelial renal cells evaluated by a microcalorimetric technique.

    PubMed

    Xie, Y; DePierre, J W; Nässberger, L

    2000-09-01

    In the present study we have developed a microcalorimetric procedure which allows convenient investigation of biocompatibility in a microsystem. We examined the biocompatibility of a porcine renal epithelial tubule cell line LLC-PK1 and a human primary renal epithelial tubule cell (RPTEC) with microplates composed of three different materials, i.e. Thermanox, transparent film and titanium. All three materials showed equal biocompatibility with LLC-PK1 cells, judging from the attainment of steady-state power curves and the same rate of heat production per cell (2.5 microW / microg DNA). The human renal cells were poorly biocompatible with the Thermanox and transparent film. However, on titanium the RPTEC cell did adhere, as demonstrated by a steady-state power curve. The human cells also showed a higher metabolic activity (3.0 microW / microg DNA), than did LLC-PK1 cells cultured on the same type of microplates. In research on biocompatibility there is a need for alternatives to experimental animal investigations. The present technique allows studies of cellular interactions with different biomaterials in a rapid and standardized manner and may therefore prove to be a useful screening procedure. PMID:15348389

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

    DOE PAGESBeta

    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 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

  17. 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

  18. Inline chemical process analysis in micro-plants based on thermoelectric flow and impedimetric sensors

    NASA Astrophysics Data System (ADS)

    Jacobs, T.; Kutzner, C.; Kropp, M.; Brokmann, G.; Lang, W.; Steinke, A.; Kienle, A.; Hauptmann, P.

    2010-10-01

    In micro-plants, as used in chemical micro-process engineering, an integrated inline analytics is regarded as an important factor for the development and optimization of chemical processes. Up to now, there is a lack of sensitive, robust and low-priced micro-sensors for monitoring mixing and chemical conversion in micro-fluidic channels. In this paper a novel sensor system combining an impedimetric sensor and a novel pressure stable thermoelectric flow sensor for monitoring chemical reactions in micro-plants is presented. The CMOS-technology-based impedimetric sensor mainly consists of two capacitively coupled interdigital electrodes on a silicon chip. The thermoelectric flow sensor consists of a heater in between two thermopiles on a perforated membrane. The pulsed and constant current feeds of the heater were analyzed. Both sensors enable the analysis of chemical conversion by means of changes in the thermal and electrical properties of the liquid. The homogeneously catalyzed synthesis of n-butyl acetate as a chemical model system was studied. Experimental results revealed that in an overpressure regime, relative changes of less than 1% in terms of thermal and electrical properties can be detected. Furthermore, the transition from one to two liquid phases accompanied by the change in slug flow conditions could be reproducibly detected.

  19. 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.

  20. AgI microplate monocrystals with polar {0001} facets: spontaneous photocarrier separation and enhanced photocatalytic activity.

    PubMed

    Kuang, Qin; Zheng, Xiaoli; Yang, Shihe

    2014-02-24

    Elucidating the facet-dependent photocatalytic activity of semiconductor photocatalysts is important in improving the overall efficiency of photocatalysis. Furthermore, combining facet control with selective deposition of oxidation and/or reduction cocatalysts on specific faces of semiconductor photocatalysts is potentially an effective strategy to synergistically optimize the functionality of photocatalysts. In the present study, high-purity wurtzite-type β-AgI platelet microcrystals with polar {0001} facets were prepared by a facile polyvinylpyrrolidone-assisted precipitation reaction. The polar-faceted AgI microplates were used as archetypes to demonstrate preferential diametric migration (i.e., effective separation) of photogenerated electrons and holes along the c axis. Such vectorial electron-hole separation stems from the asymmetric surface structures, which give rise to distinct photoexcited reaction behaviors on the ±(0001) polar facets of wurtzite-type semiconductors. Furthermore, on selective deposition of Ag and MnOx (1.5microplates in degrading organic pollutants was dramatically enhanced thanks to the broad light-absorption range, strong dye-adsorption ability, and effective spatial separation of photocarriers. PMID:24449437

  1. Self-assembly of gold nanorods into vertically aligned, rectangular microplates with a supercrystalline structure.

    PubMed

    Xiao, Junyan; Li, Zhe; Ye, Xiaozhou; Ma, Yurong; Qi, Limin

    2014-01-21

    Vertically aligned, supercrystalline microplates with a well-defined rectangular shape were fabricated in a large area through self-assembly of gold nanorods by a novel bulk solution evaporation method. This evaporative self-assembly strategy involving continuous movement of the contact line can prevent the coffee-ring effect, thus allowing uniform deposition of discrete GNR superstructures over a large area and favoring the formation of GNR supercrystals with geometrically symmetric shapes. A mechanism based on the continuing nucleation and growth of smectic GNR superstructures accompanying the movement of the contact line was put forward for the formation of the unique GNR supercrystal arrays. Based on this mechanism, a micropatterned substrate was designed to control the nucleation location and growth direction, leading to the spontaneous self-assembly of nearly parallel arrays of vertically aligned, supercrystalline microplates of GNRs. The obtained rectangular-plate-shaped GNR supercrystals exhibited interesting anisotropic optical reflection properties, which were revealed by polarized light microscopy. PMID:24292548

  2. An analytical model for squeeze-film damping of perforated torsional microplates resonators.

    PubMed

    Li, Pu; Fang, Yuming

    2015-01-01

    Squeeze-film damping plays a significant role in the performance of micro-resonators because it determines their quality factors. Perforations in microstructures are often used to control the squeeze-film damping in micro-resonators. To model the perforation effects on the squeeze-film damping, many analytical models have been proposed, however, most of the previous models have been concerned with the squeeze-film damping due to the normal motion between the perforated vibrating plate and a fixed substrate, while there is a lack of works that model the squeeze-film damping of perforated torsion microplates, which are also widely used in MEMS devices. This paper presents an analytical model for the squeeze-film damping of perforated torsion microplates. The derivation in this paper is based on a modified Reynolds equation that includes compressibility and rarefaction effects. The pressure distribution under the vibrating plate is obtained using the double sine series. Closed-form expressions for the stiffness and the damping coefficients of the squeeze-film are derived. The accuracy of the model is verified by comparing its results with the finite element method (FEM) results and the experimental results available in the literature. The regime of validity and limitations of the present model are assessed. PMID:25815453

  3. 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

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

    ERIC Educational Resources Information Center

    Binder, Katherine S.; Lee, Cheryl S.

    2012-01-01

    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)…

  5. 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…

  6. 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

  7. 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…

  8. 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…

  9. 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…

  10. Native American Adult Reader. Teacher's Guide.

    ERIC Educational Resources Information Center

    King, Lovern Root, Ed.

    Behavioral objectives and supplemental student activities are featured in this teacher's guide for using the Native American Adult Readers in basic education/GED programs. The guide provides the rationale for developing the readers, which contain a variety of brief articles and essays and are designed to provide meaningful material based on Native…

  11. 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…

  12. Advanced Hindi Reader in the Social Sciences.

    ERIC Educational Resources Information Center

    Vatuk, Ved Prakash

    This reader contains 25 selections in standard Hindi by recognized authorities in the major fields of social science; namely sociology, anthropology, folklore, economics, and political science. The writings, evenly divided both in content and style, are intended to give the reader a broad perspective of Indian culture. A 128-page Hindi-English…

  13. 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…

  14. UNDERSTANDING AND HELPING THE RETARDED READER.

    ERIC Educational Resources Information Center

    STRANG, RUTH, ED.

    THE PROCEEDINGS OF A 1962 STATEWIDE ARIZONA CONFERENCE ON READING DEVELOPMENT AND READING DIFFICULTIES INCLUDE 15 PAPERS. ARTICLES ON THE ABLE RETARDED READER ARE "UNDERSTANDING THE ABLE RETARDED READER" BY HELEN M. ROBINSON, AND "CLASSROOM PROCEDURES" BY ROSEMARY YOAKUM. PAPERS ON EMOTIONALLY DISTURBED CHILDREN ARE "IDENTIFICATION OF EMOTIONAL…

  15. Accelerated Reader. What Works Clearinghouse Intervention Report

    ERIC Educational Resources Information Center

    What Works Clearinghouse, 2009

    2009-01-01

    "Accelerated Reader" is a computer-based reading management system designed to complement an existing classroom literacy program for grades pre-K-12. It is designed to increase the amount of time students spend reading independently. Students choose reading-level appropriate books or short stories for which Accelerated Reader tests are available…

  16. Verbal Reports: How Readers Process Unfamiliar Text.

    ERIC Educational Resources Information Center

    Marr, Mary Beth

    With the use of verbal report strategies, a study was conducted to examine (1) the types of comprehension strategies readers use to process familiar and less familiar texts and (2) the differential use of think aloud strategies by average and below average readers. Subjects were 15 tenth grade male students in upstate New York. Two weeks prior to…

  17. 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…

  18. Practicum Training for Teachers of Struggling Readers

    ERIC Educational Resources Information Center

    Morris, Darrell

    2011-01-01

    Teachers who work with struggling beginning readers need a supervised training experience that leads them to understand both how reading ability develops and how to adapt instruction to meet the needs of individual children. The practicum, in which a teacher works with one struggling reader under the supervision of an experienced and expert…

  19. 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…

  20. 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…

  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. Secondary School Students' Opinions about Readers' Theatre

    ERIC Educational Resources Information Center

    Karabag, S. Gulin

    2015-01-01

    In this article, a teaching strategy which not only blends yesterday and today in a meaningful way but also powerfully integrates literacy and history will be examined. Firstly Readers' Theatre as a technique will be introduced. Secondly, the usage guidelines of Readers' Theatre will be presented. Finally the opinions of secondary school students…

  3. Technology: News Readers and Other Handy Utilities

    ERIC Educational Resources Information Center

    Van Horn, Royal

    2004-01-01

    In this article, the author discusses how there are advantages and disadvantages to using an Internet News Reader instead of a Web browser. The major advantage is that one can read the headlines and short summaries of news articles from dozens of sources quickly. Another advantage the author points out to news readers is that one gets a short…

  4. 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 IRAQI KURDISTAN, THESE 12…

  5. The Portrayal of Reader-Writer Conferencing.

    ERIC Educational Resources Information Center

    Della-Piana, Gabriel M.

    Reader-writer conferencing was examined as an alternative or complement to the direct assessment of writing. Criteria guiding development of a framework for analysis of reader-writer conferencing were summarized as: (1) achievability; (2) transfer to other domains of writing; (3) importance of the outcomes; (4) inter-scorer agreement; and (5)…

  6. 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" that may impede the…

  7. Chemical Inhibitors of Epigenetic Methyllysine Reader Proteins.

    PubMed

    Milosevich, Natalia; Hof, Fraser

    2016-03-22

    Protein methylation is a common post-translational modification with diverse biological functions. Methyllysine reader proteins are increasingly a focus of epigenetics research and play important roles in regulating many cellular processes. These reader proteins are vital players in development, cell cycle regulation, stress responses, oncogenesis, and other disease pathways. The recent emergence of a small number of chemical inhibitors for methyllysine reader proteins supports the viability of these proteins as targets for drug development. This article introduces the biochemistry and biology of methyllysine reader proteins, provides an overview of functions for those families of readers that have been targeted to date (MBT, PHD, tudor, and chromodomains), and reviews the development of synthetic agents that directly block their methyllysine reading functions. PMID:26650180

  8. Caveats and limitations of plate reader-based high-throughput kinetic measurements of intracellular calcium levels

    SciTech Connect

    Heusinkveld, Harm J.; Westerink, Remco H.S.

    2011-08-15

    Calcium plays a crucial role in virtually all cellular processes, including neurotransmission. The intracellular Ca{sup 2+} concentration ([Ca{sup 2+}]{sub i}) is therefore an important readout in neurotoxicological and neuropharmacological studies. Consequently, there is an increasing demand for high-throughput measurements of [Ca{sup 2+}]{sub i}, e.g. using multi-well microplate readers, in hazard characterization, human risk assessment and drug development. However, changes in [Ca{sup 2+}]{sub i} are highly dynamic, thereby creating challenges for high-throughput measurements. Nonetheless, several protocols are now available for real-time kinetic measurement of [Ca{sup 2+}]{sub i} in plate reader systems, though the results of such plate reader-based measurements have been questioned. In view of the increasing use of plate reader systems for measurements of [Ca{sup 2+}]{sub i} a careful evaluation of current technologies is warranted. We therefore performed an extensive set of experiments, using two cell lines (PC12 and B35) and two fluorescent calcium-sensitive dyes (Fluo-4 and Fura-2), for comparison of a linear plate reader system with single cell fluorescence microscopy. Our data demonstrate that the use of plate reader systems for high-throughput real-time kinetic measurements of [Ca{sup 2+}]{sub i} is associated with many pitfalls and limitations, including erroneous sustained increases in fluorescence, limited sensitivity and lack of single cell resolution. Additionally, our data demonstrate that probenecid, which is often used to prevent dye leakage, effectively inhibits the depolarization-evoked increase in [Ca{sup 2+}]{sub i}. Overall, the data indicate that the use of current plate reader-based strategies for high-throughput real-time kinetic measurements of [Ca{sup 2+}]{sub i} is associated with caveats and limitations that require further investigation. - Research Highlights: > The use of plate readers for high-throughput screening of intracellular

  9. A Microplate-Based System as In Vitro Model of Biofilm Growth and Quantification.

    PubMed

    Vandecandelaere, Ilse; Van Acker, Heleen; Coenye, Tom

    2016-01-01

    We describe a 96-well microtiter plate-based system as an in vitro model for biofilm formation and quantification. Although in vitro assays are artificial systems and thus significantly differ from in vivo conditions, they represent an important tool to evaluate biofilm formation and the effect of compounds on biofilms. Stainings to evaluate the amount of biomass (crystal violet staining) and the number of metabolically active cells (resazurin assay) are discussed and specific attention is paid to the use of this model to quantify persisters in sessile populations. PMID:26468099

  10. 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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  12. 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...

  13. 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...

  14. 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

  15. 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.

  16. Application of Imaging-Based Assays in Microplate Formats for High-Content Screening.

    PubMed

    Fogel, Adam I; Martin, Scott E; Hasson, Samuel A

    2016-01-01

    The use of multiparametric microscopy-based screens with automated analysis has enabled the large-scale study of biological phenomena that are currently not measurable by any other method. Collectively referred to as high-content screening (HCS), or high-content analysis (HCA), these methods rely on an expanding array of imaging hardware and software automation. Coupled with an ever-growing amount of diverse chemical matter and functional genomic tools, HCS has helped open the door to a new frontier of understanding cell biology through phenotype-driven screening. With the ability to interrogate biology on a cell-by-cell basis in highly parallel microplate-based platforms, the utility of HCS continues to grow as advancements are made in acquisition speed, model system complexity, data management, and analysis systems. This chapter uses an example of screening for genetic factors regulating mitochondrial quality control to exemplify the practical considerations in developing and executing high-content campaigns. PMID:27317002

  17. Antimicrobial susceptibility of standard strains of nontuberculous mycobacteria by microplate Alamar Blue assay.

    PubMed

    Li, Guilian; Lian, Lu-Lu; Wan, Li; Zhang, Jingrui; Zhao, Xiuqin; Jiang, Yi; Zhao, Li-Li; Liu, Haican; Wan, Kanglin

    2013-01-01

    In this study, 24 standard nontuberculous mycobacteria (NTM) species strains including 12 slowly growing mycobacteria strains and 12 rapidly growing mycobacteria strains were subjected to drug susceptibility testing using microplate Alamar Blue assay-based 7H9 broth. The most active antimicrobial agents against the 24 NTM strains were streptomycin, amikacin, the fluoroquinolones, and the tetracyclines. Mycobacterium chelonae, Mycobacterium abscessus, Mycobacterium bolletii, and Mycobacterium simiae are resistant to most antimicrobial agents. The susceptibility results of this study from 24 NTM standard strains can be referenced by clinicians before susceptibility testing for clinical isolates is performed or when conditions do not allow for susceptibility testing. The application of broth-based methods is recommended by the Clinical and Laboratory Standards Institute, and the documentation of the susceptibility patterns of standard strains of mycobacteria can improve the international standardization of susceptibility testing methods. PMID:24386332

  18. 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. PMID:27542445

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

    NASA Astrophysics Data System (ADS)

    Farokhi, Hamed; Ghayesh, Mergen H.

    2015-12-01

    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.

  20. Highly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification

    PubMed Central

    Moudjou, Mohammed; Sibille, Pierre; Fichet, Guillaume; Reine, Fabienne; Chapuis, Jérôme; Herzog, Laetitia; Jaumain, Emilie; Laferrière, Florent; Richard, Charles-Adrien; Laude, Hubert; Andréoletti, Olivier; Rezaei, Human; Béringue, Vincent

    2013-01-01

    ABSTRACT Measurements of the presence of prions in biological tissues or fluids rely more and more on cell-free assays. Although protein misfolding cyclic amplification (PMCA) has emerged as a valuable, sensitive tool, it is currently hampered by its lack of robustness and rapidity for high-throughput purposes. Here, we made a number of improvements making it possible to amplify the maximum levels of scrapie prions in a single 48-h round and in a microplate format. The amplification rates and the infectious titer of the PMCA-formed prions appeared similar to those derived from the in vivo laboratory bioassays. This enhanced technique also amplified efficiently prions from different species, including those responsible for human variant Creutzfeldt-Jakob disease. This new format should help in developing ultrasensitive, high-throughput prion assays for cognitive, diagnostic, and therapeutic applications. PMID:24381300

  1. Adaptation of Microplate-based Respirometry for Hippocampal Slices and Analysis of Respiratory Capacity

    PubMed Central

    Schuh, Rosemary A.; Clerc, Pascaline; Hwang, Hyehyun; Mehrabian, Zara; Bittman, Kevin; Chen, Hegang; Polster, Brian M.

    2011-01-01

    Multiple neurodegenerative disorders are associated with altered mitochondrial bioenergetics. Although mitochondrial O2 consumption is frequently measured in isolated mitochondria, isolated synaptic nerve terminals (synaptosomes), or cultured cells, the absence of mature brain circuitry is a remaining limitation. Here we describe the development of a method that adapts the Seahorse Extracellular Flux Analyzer (XF24) for the microplate-based measurement of hippocampal slice O2 consumption. As a first evaluation of the technique, we compared whole slice bioenergetics to previous measurements made with synaptosomes or cultured neurons. We found that mitochondrial respiratory capacity and O2 consumption coupled to ATP synthesis could be estimated in cultured or acute hippocampal slices with preserved neural architecture. Mouse organotypic hippocampal slices oxidizing glucose displayed mitochondrial O2 consumption that was well-coupled, as determined by the sensitivity to the ATP synthase inhibitor oligomycin. However stimulation of respiration by uncoupler was modest (<120% of basal respiration) compared to previous measurements in cells or synaptosomes, although enhanced slightly (to ~150% of basal respiration) by the acute addition of the mitochondrial complex I-linked substrate pyruvate. These findings suggest a high basal utilization of respiratory capacity in slices and a limitation of glucose-derived substrate for maximal respiration. The improved throughput of microplate-based hippocampal respirometry over traditional O2 electrode-based methods is conducive to neuroprotective drug screening. When coupled with cell type-specific pharmacology or genetic manipulations, the ability to efficiently measure O2 consumption from whole slices should advance our understanding of mitochondrial roles in physiology and neuropathology. PMID:21520220

  2. 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

  3. Crustal Structure of the Yakutat Microplate: Constraints from STEEP Wide-angle Seismic Data

    NASA Astrophysics Data System (ADS)

    Christeson, G. L.; van Avendonk, H.; Gulick, S. P.; Worthington, L.; Pavlis, T.

    2008-12-01

    In Fall 2008 we will conduct a seismic program focusing on the Yakutat microplate. As part of this study we plan to acquire two wide-angle profiles: an onshore-offshore northwest-southeast oriented profile extending from the Bering glacier onto the continental shelf and across the Dangerous River Zone, and an offshore northeast-southwest oriented profile extending from the ocean basin across the Transition fault and into Yakutat Bay. The sound source will be the R/V Langseth's tuned 6600 cu. in., 36 air gun array. Ocean bottom seismometers will be positioned at ~15 km spacing, and Texan seismometers at 1-4 km spacing across the Bering Glacier. Coincident deep-penetrating seismic reflection data will be acquired on the marine portion of both profiles using a 8-km, 640-channel solid hydrophone streamer. Existing models for the Yakutat microplate disagree as to whether it is a continental fragment attached to normal oceanic crust or an oceanic plateau, and if the deep structure changes from west to east across the Dangerous River Zone. In the continental fragment model uplift is concentrated along crustal-scale thrust faulting at the ocean crust boundary (Dangerous River Zone?) resulting in focused and rapid erosion. In the oceanic plateau model more distributed, regional uplift is expected which will produce widespread exhumation with net erosion potentially coupled with glacial cycles. Thus distinguishing between these models, which we expect to accomplish with our planned seismic program, is vital for linking tectonics to erosion on both spatial and temporal scales.

  4. Two Variants of a High-Throughput Fluorescent Microplate Assay of Polysaccharide Endotransglycosylases.

    PubMed

    Kováčová, Kristína; Farkaš, Vladimír

    2016-04-01

    Polysaccharide endotransglycosylases (PETs) are the cell wall-modifying enzymes of fungi and plants. They catalyze random endo-splitting of the polysaccharide donor molecule and transfer of the newly formed reducing sugar residue to the nonreducing end of an acceptor molecule which can be a polysaccharide or an oligosaccharide. Owing to their important role in the cell wall formation, the inhibition of PETs represents an attractive strategy in the fight against fungal infections. We have elaborated two variants of a versatile high-throughput microplate fluorimetric assay that could be used for effective identification of PETs and screening of their inhibitors. Both assays use the respective polysaccharides as the donors and sulforhodamine-labeled oligosaccharides as the acceptors but differ from each other by mode of how the labeled polysaccharide products of transglycosylation are separated from the unreacted oligosaccharide acceptors. In the first variant, the reactions take place in a layer of agar gel laid on the bottoms of the wells of a microtitration plate. After the reaction, the high-Mr transglycosylation products are precipitated with 66 % ethanol and retained within the gel while the low-Mr products and the unreacted acceptors are washed out. In the second variant, the donor polysaccharides are adsorbed to the surface of a microplate well and remain adsorbed there also after becoming labeled in the course of the transglycosylation reaction whereas the unused low-Mr acceptors are washed out. As a proof of versatility, assays of heterologously expressed transglycosylases ScGas1, ScCrh1, and ScCrh2 from the yeast Saccharomyces cerevisiae, CaPhr1 and CaPhr2 from Candida albicans, and of a plant xyloglucan endotransglycosylase (XET) are demonstrated. PMID:26754421

  5. Novel high-sensitivity fluorescence polarization reader

    NASA Astrophysics Data System (ADS)

    Hoyt, Clifford C.; Levenson, Richard M.; Banks, Peter

    2001-05-01

    We have developed a new fluorescence polarization (FP) reader suitable for high-throughput screening (HST) and ultra-HTS whose assay-performance and sample-throughput are both considerably improved over present state-of-the-art instrumentation. The SymmetryTM reader possesses a number of features that differ from conventional HTS FP readers. These include: laser-based excitation, liquid crystal polarization optics that rapidly and accurately measure polarization states; and CCD detectors to capture emission from multiple wells. We show that the performance in assays relevant to the drug discovery process, such as G- protein coupled receptor-based assays, is significantly enhanced due to a dramatic improvement in precision. Furthermore, the CCD-detection system used can substantially improve sample throughput compared to sequential readers while maintaining high performance.

  6. Fluorescence lifetime plate reader: Resolution and precision meet high-throughput

    NASA Astrophysics Data System (ADS)

    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.

  7. Fluorescence lifetime plate reader: Resolution and precision meet high-throughput

    PubMed Central

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

    2014-01-01

    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. PMID:25430092

  8. 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

  9. 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.

  10. Damping constant estimation in magnetoresistive readers

    NASA Astrophysics Data System (ADS)

    Stankiewicz, Andrzej; Hernandez, Stephanie

    2015-05-01

    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.

  11. "True Wizardry": Readers Theatre in the Classroom. PEN 79.

    ERIC Educational Resources Information Center

    Robertson, Marion E.

    This article describes Readers Theater (a form of group storytelling in which two or more readers present a piece of literature by reading aloud from hand-held scripts) and advocates its use in the classroom. The paper's seven sections are as follows: (1) What is Readers Theater; (2) Readers Theater and Its Conventions (discussing scripts,…

  12. "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…

  13. 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…

  14. Development of Magnetization Measurement Devices Using Micro-dc-SQUIDs and a Sr_2RuO_4 Microplate

    NASA Astrophysics Data System (ADS)

    Nago, Y.; Shinozaki, T.; Tsuchiya, S.; Ishiguro, R.; Kashiwaya, H.; Kashiwaya, S.; Nomura, S.; Kono, K.; Takayanagi, H.; Maeno, Y.

    2016-05-01

    We developed high-sensitivity magnetization measurement devices composed of micro-dc-SQUIDs and a superconducting Sr_2RuO_4 microplate, aiming to investigate novel magnetic properties related to a spin-triplet chiral p-wave superconductor with a mesoscopic size. Micron-sized dc-SQUID was fabricated by thin Al electrodes, and the SQUID structure was improved to prevent magnetic fluxes from intruding into SQUID electrodes. A Sr_2RuO_4 superconducting microplate was fabricated into the size as small as the SQUID loop using a focused ion beam and directly mounted on the SQUID with precise positioning for high-sensitivity magnetization measurements. In the preliminary magnetization measurements of this device, we observed vortices trapped into the plate and thus the lower critical field. The improved magnetization measurement device developed to exclude undesirable flux intrusion successfully enabled high-sensitivity detection of quantized vortex.

  15. Microplate MPN-enumeration of monocyclic- and dicyclic-aromatic hydrocarbon degraders via substrate phase-partitioning.

    PubMed

    Johnsen, Anders R; Henriksen, Sidsel

    2009-07-01

    The high aqueous solubility of monoaromatic and some diaromatic oil components may hinder classical growth-based MPN enumeration of bacterial mono- and di-aromatics degraders because these aromatics are toxic in high concentrations. We developed a microplate MPN method for the enumeration of toluene-, xylene-, naphthalene-, biphenyl- and benzothiophene-degraders on the basis of phase-partitioning of substrate between a biologically inert organic phase and an aqueous mineral salt medium. This way, it was possible to maintain non-toxic, aqueous concentrations in the microplate wells. Depletion of aqueous aromatics by growth of the degraders was prevented by the continuous solubilization of aromatics from the silicone phase. The method was validated by MPN enumerating degrader cultures both with phase-partitioned aromatics and with tryptic soy broth as carbon sources. The applicability of the method was demonstrated by MPN-enumerating mono- and di-aromatic degraders in soils of varying hydrocarbon pre-exposure. PMID:19043785

  16. A New Look at Spreading in Iceland: Propagating Rifts, Migrating Transform Faults, and Microplate Tectonics

    NASA Astrophysics Data System (ADS)

    Karson, J.; Horst, A. J.; Nanfito, A.

    2011-12-01

    Iceland has long been used as an analog for studies of seafloor spreading. Despite its thick (~25 km) oceanic crust and subaerial lavas, many features associated with accretion along mid-ocean ridge spreading centers, and the processes that generate them, are well represented in the actively spreading Neovolcanic Zone and deeply glaciated Tertiary crust that flanks it. Integrated results of structural and geodetic studies show that the plate boundary zone on Iceland is a complex array of linked structures bounding major crustal blocks or microplates, similar to oceanic microplates. Major rift zones propagate N and S from the hotspot centered beneath the Vatnajökull icecap in SE central Iceland. The southern propagator has extended southward beyond the South Iceland Seismic Zone transform fault to the Westman Islands, resulting in abandonment of the Eastern Rift Zone. Continued propagation may cause abandonment of the Reykjanes Ridge. The northern propagator is linked to the southern end of the receding Kolbeinsey Ridge to the north. The NNW-trending Kerlingar Pseudo-fault bounds the propagator system to the E. The Tjörnes Transform Fault links the propagator tip to the Kolbeinsey Ridge and appears to be migrating northward in incremental steps, leaving a swath of deformed crustal blocks in its wake. Block rotations, concentrated mainly to the west of the propagators, are clockwise to the N of the hotspot and counter-clockwise to the S, possibly resulting in a component of NS divergence across EW-oriented rift zones. These rotations may help accommodate adjustments of the plate boundary zone to the relative movements of the N American and Eurasian plates. The rotated crustal blocks are composed of highly anisotropic crust with rift-parallel internal fabric generated by spreading processes. Block rotations result in reactivation of spreading-related faults as major rift-parallel, strike-slip faults. Structural details found in Iceland can help provide information

  17. Microplitis demolitor Bracovirus Proviral Loci and Clustered Replication Genes Exhibit Distinct DNA Amplification Patterns during Replication

    PubMed Central

    Simmonds, Tyler J.; Thomas, Sarah A.; Strand, Michael R.

    2015-01-01

    ABSTRACT Polydnaviruses are large, double-stranded DNA viruses that are beneficial symbionts of parasitoid wasps. Polydnaviruses in the genus Bracovirus (BVs) persist in wasps as proviruses, and their genomes consist of two functional components referred to as proviral segments and nudivirus-like genes. Prior studies established that the DNA domains where proviral segments reside are amplified during replication and that segments within amplified loci are circularized before packaging into nucleocapsids. One DNA domain where nudivirus-like genes are located is also amplified but never packaged into virions. We recently sequenced the genome of the braconid Microplitis demolitor, which carries M. demolitor bracovirus (MdBV). Here, we took advantage of this resource to characterize the DNAs that are amplified during MdBV replication using a combination of Illumina and Pacific Biosciences sequencing approaches. The results showed that specific nucleotide sites identify the boundaries of amplification for proviral loci. Surprisingly, however, amplification of loci 3, 4, 6, and 8 produced head-to-tail concatemeric intermediates; loci 1, 2, and 5 produced head-to-head/tail-to-tail concatemers; and locus 7 yielded no identified concatemers. Sequence differences at amplification junctions correlated with the types of amplification intermediates the loci produced, while concatemer processing gave rise to the circularized DNAs that are packaged into nucleocapsids. The MdBV nudivirus-like gene cluster was also amplified, albeit more weakly than most proviral loci and with nondiscrete boundaries. Overall, the MdBV genome exhibited three patterns of DNA amplification during replication. Our data also suggest that PacBio sequencing could be useful in studying the replication intermediates produced by other DNA viruses. IMPORTANCE Polydnaviruses are of fundamental interest because they provide a novel example of viruses evolving into beneficial symbionts. All polydnaviruses are

  18. Effects of van der Waals Force and Thermal Stresses on Pull-in Instability of Clamped Rectangular Microplates

    PubMed Central

    Batra, Romesh C.; Porfiri, Maurizio; Spinello, Davide

    2008-01-01

    We study the influence of von Kármán nonlinearity, van der Waals force, and thermal stresses on pull-in instability and small vibrations of electrostatically actuated microplates. We use the Galerkin method to develop a tractable reduced-order model for electrostatically actuated clamped rectangular microplates in the presence of van der Waals forces and thermal stresses. More specifically, we reduce the governing two-dimensional nonlinear transient boundary-value problem to a single nonlinear ordinary differential equation. For the static problem, the pull-in voltage and the pull-in displacement are determined by solving a pair of nonlinear algebraic equations. The fundamental vibration frequency corresponding to a deflected configuration of the microplate is determined by solving a linear algebraic equation. The proposed reduced-order model allows for accurately estimating the combined effects of van der Waals force and thermal stresses on the pull-in voltage and the pull-in deflection profile with an extremely limited computational effort.

  19. 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.

  20. What Eye Movements Reveal about Deaf Readers

    PubMed Central

    Bélanger, Nathalie N.; Rayner, Keith

    2014-01-01

    Levels of illiteracy in the deaf populations around the world have been extremely high for decades and much higher than the illiteracy levels found in the general population. Research has mostly focused on deaf readers’ difficulties rather than on their strengths, which can then inform reading education. Deaf readers are a unique population. They process language and the world surrounding them mostly via the visual channel and this greatly affects how they read or might learn to read. The study of eye movements in reading provides highly sophisticated information about how words and sentences are processed and our research with deaf readers reveals the importance of their uniqueness. PMID:26594098

  1. Cloning and expression profile of ionotropic receptors in the parasitoid wasp Microplitis mediator (Hymenoptera: Braconidae).

    PubMed

    Wang, Shan-Ning; Peng, Yong; Lu, Zi-Yun; Dhiloo, Khalid Hussain; Zheng, Yao; Shan, Shuang; Li, Rui-Jun; Zhang, Yong-Jun; Guo, Yu-Yuan

    2016-07-01

    Ionotropic receptors (IRs) mainly detect the acids and amines having great importance in many insect species, representing an ancient olfactory receptor family in insects. In the present work, we performed RNAseq of Microplitis mediator antennae and identified seventeen IRs. Full-length MmedIRs were cloned and sequenced. Phylogenetic analysis of the Hymenoptera IRs revealed that ten MmedIR genes encoded "antennal IRs" and seven encoded "divergent IRs". Among the IR25a orthologous groups, two genes, MmedIR25a.1 and MmedIR25a.2, were found in M. mediator. Gene structure analysis of MmedIR25a revealed a tandem duplication of IR25a in M. mediator. The tissue distribution and development specific expression of the MmedIR genes suggested that these genes showed a broad expression profile. Quantitative gene expression analysis showed that most of the genes are highly enriched in adult antennae, indicating the candidate chemosensory function of this family in parasitic wasps. Using immunocytochemistry, we confirmed that one co-receptor, MmedIR8a, was expressed in the olfactory sensory neurons. Our data will supply fundamental information for functional analysis of the IRs in parasitoid wasp chemoreception. PMID:27208597

  2. Nonlinear size-dependent dynamics of an imperfect shear deformable microplate

    NASA Astrophysics Data System (ADS)

    Farokhi, Hamed; Ghayesh, Mergen H.

    2016-01-01

    The nonlinear size-dependent dynamics of a geometrically imperfect shear deformable microplate is investigated based on the modified couple stress theory. An initial imperfection in the out-of-plane direction is taken into account. The third-order shear deformation theory is employed to model the system by retaining in-plane displacements and inertia. The governing equations of motion of the system are obtained employing an energy method through use of the Lagrange equations, which upon employing an assumed-mode technique, yields a set of second-order nonlinear ordinary differential equations with coupled terms. A high-dimensional discretised system is considered and made double-dimensional via a change of variables in order to obtain a set of first-order nonlinear ordinary differential equations. The resulting equations are solved using a direct time-integration technique, resulting in time-dependent generalised coordinates for the in-plane and out-of-plane displacements and two rotations. From these generalised coordinates, phase-plane portraits and fast Fourier transforms (FFTs) are also obtained. Moreover, the frequency-response and force-response curves of the system are obtained using a continuation technique; stability analysis is conducted via the Floquet theory. The effect of the initial imperfection as well as the length-scale parameter on the system response is also examined.

  3. Identification and Expression Analysis of Putative Chemosensory Receptor Genes in Microplitis mediator by Antennal Transcriptome Screening

    PubMed Central

    Wang, Shan-Ning; Peng, Yong; Lu, Zi-Yun; Dhiloo, Khalid Hussain; Gu, Shao-Hua; Li, Rui-Jun; Zhou, Jing-Jiang; Zhang, Yong-Jun; Guo, Yu-Yuan

    2015-01-01

    Host-seeking, ovipositional behavior and mating of insects are controlled mainly by odor perception through sensory organs such as antennae. Antennal chemoreception is extremely important for insect survival. Several antennal chemosensory receptors are involved in mediating the odor detection in insects, especially the odorant receptors (ORs) and ionotropic receptors (IRs), to ensure the specificity of the olfactory sensory neuron responses. In the present study, we identified the chemosensory receptor gene repertoire of the parasitoid wasp Microplitis mediator, a generalist endoparasitoid that infests more than 40 types of Lepidopterous larvae and is widely distributed in the Palaearctic region. By transcriptome sequencing of male and female antennae we identified 60 candidate odorant receptors, six candidate ionotropic receptors and two gustatory receptors in M. mediator. The full-length sequences of these putative chemosensory receptor genes were obtained by using the rapid amplification of cDNA ends PCR (RACE-PCR) method. We also conducted reverse transcription PCR (RT-PCR) combined with real-time quantitative PCR (qPCR) for investigating the expression profiles of these chemosensory receptor genes in olfactory and non-olfactory tissues. The tissue- and sex-biased expression patterns may provide insights into the roles of the chemosensory receptor in M. mediator. Our findings support possible future study of the chemosensory behavior of M. mediator at the molecular level. PMID:26078716

  4. Development of a fluorometric microplate antiadhesion assay using uropathogenic Escherichia coli and human uroepithelial cells.

    PubMed

    Kimble, Lindsey L; Mathison, Bridget D; Kaspar, Kerrie L; Khoo, Christina; Chew, Boon P

    2014-05-23

    A fluorometric microplate assay has been developed to determine Escherichia (E.) coli adhesion to uroepithelial cells (UEC). P-fimbriated E. coli were labeled with BacLight Green and preincubated 30 min with human urine or standard. Fluorescent-E. coli were added to UEC in mircoplates at a 400:1 ratio, incubated 1 h, and washed, and the fluorescence intensity was measured. Specific labeling and adherence were confirmed by flow cytometry. A myricetin (1) standard curve (0-30 μg/mL) was developed; the lower limit of detection was 0.1 μg/mL, and half-maximal inhibitory concentration was 0.88 μg/mL (intra- and interassay coefficients of variance were <10% and <15%, respectively). Vaccinium macrocarpon (cranberry) extracts, quercetin (2), and procyanidins B1 (3), B2 (4), and C1 (5) showed similar inhibition. Antiadhesion activity of urine samples from subjects (n = 12) consuming placebo or V. macrocarpon beverage determined using this assay was positively correlated (R(2) = 0.78; p < 0.01) with a radiolabeled-E. coli assay. PMID:24749980

  5. 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

  6. Knockdown of microplitis mediator odorant receptor involved in the sensitive detection of two chemicals.

    PubMed

    Li, Ke-Ming; Ren, Li-Yan; Zhang, Yong-Jun; Wu, Kong-Ming; Guo, Yu-Yuan

    2012-03-01

    Odorant receptors are thought to play critical roles in the perception of chemosensory stimuli by insects. The primary method to address the functions of odorant receptors in insects is to use in vitro binding assays between the receptors and potential chemical stimuli. We injected MmedOrco dsRNA into the abdominal cavity of a braconid wasp, Microplitis mediator, and assayed for expression of this gene 72 h after treatment (RNAi). Quantitative real-time PCR demonstrated that the level of mRNA expression in MmedOrco dsRNA-treated M. mediator was significantly reduced (>90%) when compared with water-treated controls. Furthermore, electroantennogram (EAG) responses of M. mediator to two chemical attractants, nonanal and farnesene, were also reduced significantly (~70%) in RNAi-treated M. mediator when compared to controls. RNAi-treated M. mediator also responded by walking/flying at a lower rate to both chemicals when compared with controls in a Y-tube olfactometer bioassay, which provides direct evidence that MmedOrco plays an important role in perception of nonanal and farnesene in M. mediator. PMID:22402893

  7. 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. PMID:25482381

  8. Microplate biochemical determination of Russian VX: influence of admixtures and avoidance of false negative results.

    PubMed

    Prokofieva, Daria S; Jenkins, Richard O; Goncharov, Nikolay V

    2012-05-15

    Two microplate spectroscopic methods for determination of organophosphates, based on inhibition of acetylcholinesterase (AChE) activity, were further improved and evaluated for determination of the chemical weapon agent Russian VX (RVX) in aqueous solutions. The linear range of the Hestrin method (74.8-1120 pM) was 3.1-fold wider than that of the Ellman method (37.4-374 pM). Limits of detection and quantification of RVX for both methods were below the maximal allowable concentration of RVX in water-soluble washouts. One of the early products of RVX hydrolysis, N,N-diethylaminoethanethiol, like reduced glutathione, caused false negative results in the Ellman method at concentrations exceeding 10 μM; individual blanks were necessary to eliminate the effect. The Hestrin method showed greater specificity (~3 orders of magnitude) for analysis of samples containing mercaptans. A major product of RVX degradation, 2,2'-dithiobis(N,N-diethylethanamine), caused significant inhibition of AChE at concentrations of ≥0.1 mM (P<0.01) and had a false positive effect at higher concentrations (≥2 mM). For environmental monitoring of RVX, the method based on Hestrin is preferred over that based on Ellman, principally because the former method was less sensitive to interference from major admixtures and did not give rise to potentially dangerous false negative results. PMID:22381367

  9. High-throughput microplate assay for the determination of drug partition coefficients.

    PubMed

    Magalhães, Luís M; Nunes, Cláudia; Lúcio, Marlene; Segundo, Marcela A; Reis, Salette; Lima, José L F C

    2010-11-01

    Partition coefficients (K(p)) of drugs between the phospholipid bilayer and the aqueous phase provide useful information in quantitative structure-activity relationship studies. Hexadecylphosphocholine (HePC) micelles, composed of a zwitterionic hydrophilic surface and a hydrophobic core, mimic the biomembranes and have several advantages over other lipid structures to assess K(p) values. Their preparation is easy, fast and avoids the use of toxic organic solvents, and the output has fewer spectroscopic interferences. Here, we describe a high-throughput microplate protocol for assessing the K(p) of drugs using HePC micelles as membrane models and derivative spectrophotometry as the detection technique. Moreover, the time-consuming data treatment to assess K(p) values is easily performed by a dedicated Excel routine developed here and described in detail. The K(p) values of nonsteroidal anti-inflammatory drugs (acemetacin, clonixin, diclofenac and indomethacin) were determined to show the simplicity of the method and to validate this protocol, which provides K(p) values (n = 3) of two drugs in ∼ 2 h. PMID:21030957

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

    PubMed

    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

  11. Kinetic microplate bioassays for relative potency of antibiotics improved by partial Least Square (PLS) regression.

    PubMed

    Francisco, Fabiane Lacerda; Saviano, Alessandro Morais; Almeida, Túlia de Souza Botelho; Lourenço, Felipe Rebello

    2016-05-01

    Microbiological assays are widely used to estimate the relative potencies of antibiotics in order to guarantee the efficacy, safety, and quality of drug products. Despite of the advantages of turbidimetric bioassays when compared to other methods, it has limitations concerning the linearity and range of the dose-response curve determination. Here, we proposed to use partial least squares (PLS) regression to solve these limitations and to improve the prediction of relative potencies of antibiotics. Kinetic-reading microplate turbidimetric bioassays for apramacyin and vancomycin were performed using Escherichia coli (ATCC 8739) and Bacillus subtilis (ATCC 6633), respectively. Microbial growths were measured as absorbance up to 180 and 300min for apramycin and vancomycin turbidimetric bioassays, respectively. Conventional dose-response curves (absorbances or area under the microbial growth curve vs. log of antibiotic concentration) showed significant regression, however there were significant deviation of linearity. Thus, they could not be used for relative potency estimations. PLS regression allowed us to construct a predictive model for estimating the relative potencies of apramycin and vancomycin without over-fitting and it improved the linear range of turbidimetric bioassay. In addition, PLS regression provided predictions of relative potencies equivalent to those obtained from agar diffusion official methods. Therefore, we conclude that PLS regression may be used to estimate the relative potencies of antibiotics with significant advantages when compared to conventional dose-response curve determination. PMID:26971814

  12. 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…

  13. 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.

  14. 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…

  15. 38 CFR 21.150 - Reader service.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 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, 2014 CFR

    2014-07-01

    ... 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. Readers' Tellings: Narrators, Settings, Flashbacks and Comprehension

    ERIC Educational Resources Information Center

    Kucer, Stephen B.

    2010-01-01

    This research explores the impact of flashbacks and changes in settings and narrators on reader comprehension. Individually, 34 fourth graders (9 and 10 years of age), mostly with above average reading abilities (5.0), orally read the first chapter of a novel. Both publisher and readability formulae estimated the text to be at a fourth- grade…

  18. 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…

  19. 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,…

  20. The Gifted Reader: Metacognition and Comprehension Strategies.

    ERIC Educational Resources Information Center

    Wingenbach, Nancy Gard

    To examine the comprehension process employed by gifted readers and to identify the various metacognitive strategies they employ, 100 gifted student volunteers in grades 4 through 7 were administered the Iowa Tests of Basic Skills (ITBS), Reading Subtests 11 and 13. The students also completed a questionnaire to determine metacognitive awareness…

  1. Preventing the Proliferation of Problem Readers.

    ERIC Educational Resources Information Center

    Hart, M. Murlee

    The failure of many beginning reading students to become efficient or enthusiastic readers is due to mistaken views of the reading process that cause teachers to stress isolated reading skills and error-free oral reading, rather than focusing on children's ability to read with understanding. Teachers who want to teach differently must take an…

  2. Revisiting the Reader's Rudder: A Comprehension Strategy.

    ERIC Educational Resources Information Center

    Fischer, Cynthia

    2003-01-01

    Presents the structured comprehension method, a strategy that facilitates literal, inferential, and critical reading comprehension for passive readers who can decode but not comprehend. Uses the method to illustrate how other areas of students' instruction (e.g., vocabulary enhancement through morphemic analysis, use of a phonogram approach to…

  3. Reader Development Initiatives in International School Libraries.

    ERIC Educational Resources Information Center

    Tilke, Anthony

    2002-01-01

    Argues that school library policy should include support and encouragement of reading as a life-long activity. Describes a number of recent reader-development initiatives, some of which include activities involving World Book Day, sponsored by UNESCO, and the 'shadowing' schemes of the Carnegie and Kate Greenaway Medal awards. (AUTH/NB)

  4. 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...

  5. 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...

  6. Moroccan Arabic Intermediate Reader, Part II.

    ERIC Educational Resources Information Center

    Alami, Wali A.; Hodge, Carlton T., Ed.

    The first section of this companion volume to "Moroccan Arabic Intermediate Reader, Part I" (AL 002 041) presents the Arabic script version of the pre-drills in Lessons IA-IIB in that volume. The second and major section comprises 20 lessons consisting of pre-drills, texts, notes, and questions. All material in this volume appears in Arabic script…

  7. 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…

  8. 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 prominent…

  9. Tape Lessons to Accompany Intermediate Nepali Reader.

    ERIC Educational Resources Information Center

    Verma, Manindra K.

    These tape lessons follow the sequence of the intermediate Nepali Reader. There are 12 lessons each containing various types of exercises designed to increase listening, speaking, and reading skills. Each lesson contains the following types of exercises: (1) listening comprehension; (2) question answering; (3) repetition; and (4) multiple choice…

  10. Ethnographic Research: A Reader [Book Review].

    ERIC Educational Resources Information Center

    Evaluation & Research in Education, 2002

    2002-01-01

    This reader was designed to support the Open University Masters in Social Sciences program and as such includes an appendix of useful questions based on the texts used to stimulate seminar discussion. It offers insights into ethnographic research techniques and serves as a tool through which some limitations of conventional quantitative research…

  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. 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…

  13. 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…

  14. 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, humorous,…

  15. Creating a Disabled Reader: A Father's Perspective.

    ERIC Educational Resources Information Center

    Phillips, Jerry

    A father describes how the public school system made his daughter, Charlie, into a disabled reader. Charlie had experienced early reading success both at home and in kindergarten, and enjoyed reading. Placed in the low reading group in first grade, Charlie resisted the dull, structured exercises of remedial reading instruction which she received…

  16. 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…

  17. Precocious Readers: Past, Present, and Future

    ERIC Educational Resources Information Center

    Olson, Lynn A.; Evans, James R.; Keckler, Wade T.

    2006-01-01

    Precocious readers represent a small portion of children who enter school each year. Researchers have investigated the environmental characteristics, acquisition process, psycholinguistic and neuropsychological characteristics, and academic skills of these children. Despite the research findings in the area, researchers and clinicians are still…

  18. Using a Barcode Reader with Interactive Videodiscs.

    ERIC Educational Resources Information Center

    Smith, Mac; Gustafson, Kent

    1995-01-01

    Describes the use of a barcode reader, rather than a computer, to control an interactive videodisc for presentations and self-paced tutorials. Highlights include hardware and software used; remote and direct communication; a HyperCard-based program to make barcodes; and converting an existing program. (LRW)

  19. Readers' Responses to Brief News Articles.

    ERIC Educational Resources Information Center

    Goetz, Ernest T.; And Others

    A study extended the investigation of readers' imaginative processes (spontaneous imagery and emotional response) to a new genre of texts: newspaper articles. A sample of 25 articles was randomly selected from a well-defined population of naturally occurring texts (articles with one or more subheadings and three to five paragraphs before the first…

  20. 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…

  1. 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…

  2. Measuring Reader Failure at the Catalogue

    ERIC Educational Resources Information Center

    Seymour, Carol A.; Schofield, J. L.

    1973-01-01

    The Library Management Research Unit tested a survey design to measure and evaluate author catalogue use in four varying libraries. Readers were asked to note details of items not found in the catalogue and the source of the reference. Items were checked to discover the cause of failure.'' (9 references) (Author/DH)

  3. International Reports on Literacy Research: Emergent Readers

    ERIC Educational Resources Information Center

    Botzakis, Stergios, Comp.; Malloy, Jacquelynn A., Comp.

    2006-01-01

    This is a compilation of reports from international research correspondents on the topic of emergent reading. The report includes 14 separate reports on Nigeria, China, Hong Kong, Israel, Malaysia, Singapore, Australia, Estonia, Italy, Canada, Mexico, United States, Argentina, and Chile. Overall, reports on the topic of emergent readers have…

  4. Project Physics Reader 1, Concepts of Motion.

    ERIC Educational Resources Information Center

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

    As a supplement to Project Physics Unit 1, 21 articles are presented in this reader. Concepts of motion are discussed under headings: motion, motion in words, representation of movement, introducing vectors, Galileo's discussion of projectile motion, Newton's laws of dynamics, the dynamics of a golf club, report on Tait's lecture on force, and bad…

  5. 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 (IN THE FIRST 15 LESSONS), A…

  6. 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…

  7. 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."…

  8. 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…

  9. 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…

  10. 38 CFR 21.150 - Reader service.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 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...

  11. Guiding Young Readers to Multicultural Literature

    ERIC Educational Resources Information Center

    Hinton-Johnson, KaaVonia; Dickinson, Gail

    2005-01-01

    Stocking the shelves of library media centers with multicultural literature is not enough, it is important that children are helped to choose the ones that would interest them as reading about various cultures is of great benefit to young readers. The importance of accurately representing to children a multicultural society is emphasized and…

  12. 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)…

  13. 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…

  14. An Urdu Newspaper Reader. Key to an Urdu Newspaper Reader. (2 vol).

    ERIC Educational Resources Information Center

    Barker, Muhammad Abd-al-Rahman; And Others

    This Reader is the second of a four-volume series in Urdu prepared by the Institute of Islamic Studies at McGill University. (See "A Course in Urdu," ED 013 435-7; "A Reader of Modern Urdu Poetry," ED 022 163; and "An Urdu Newspaper Word Count," AL 002 059.) This volume is intended for use at the second-year level of a comprehensive program of…

  15. Measuring Ca²⁺ changes in multiwell format using the Fluorometric Imaging Plate Reader (FLIPR(®)).

    PubMed

    Marshall, Ian C B; Owen, Davina E; McNulty, Shaun

    2013-01-01

    The Fluorometric Imaging Plate Reader (FLIPR) has made a significant contribution to drug discovery programs. The key advantage of FLIPR over conventional plate readers is the ability to measure fluorescence emission from multiple wells (96 wells or 384 wells) simultaneously and with high temporal resolution. Consequently, FLIPR has been used extensively to record dynamic intracellular processes such as changes in intracellular Ca(2+) ion concentration, membrane potential, and pH. Since FLIPR is used to measure a functional response in cells, it is rapidly able to distinguish full agonists, partial agonists, and antagonists at a target of interest, making the system a valuable screening tool for interrogation of compound libraries. Automated FLIPR systems for ultra high throughput have also become available that employ integrated plate stackers, washers and specialized stages to allow users to shuttle cell and compound plates from incubators or storage magazines onto the FLIPR system itself. In this chapter generic methods for assessing intracellular Ca(2+) on the FLIPR are described. PMID:23007581

  16. 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

  17. Fluorometric microplate assay to measure glutathione S-transferase activity in insects and mites using monochlorobimane.

    PubMed

    Nauen, Ralf; Stumpf, Natascha

    2002-04-15

    Elevated levels of glutathione S-transferases (GSTs) play a major role as a mechanism of resistance to insecticides and acaricides in resistant pest insects and mites, respectively. Such compounds are either detoxicated directly via phase I metabolism or detoxicated by phase II metabolism of metabolites as formed by microsomal monooxygenases. Here we used monochlorobimane (MCB) as an artificial substrate and glutathione to determine total GST activity in equivalents of single pest insects and spider mites in a sensitive 96-well plate-based assay system by measuring the enzymatic conversion of MCB to its fluorescent bimane-glutathione adduct. The differentiation by their GST activity between several strains of the two-spotted spider mite, Tetranychus urticae (Acari: Tetranychidae), with different degrees of resistance to numerous acaricides was more sensitive with MCB compared to the commonly used substrate 1-chloro-2,4-dinitrobenzene (CDNB). Compared to an acaricide-susceptible reference strain, one field population of T. urticae showed a more than 10-fold higher GST activity measured with MCB, in contrast to a less than 2-fold higher activity when CDNB was used. Furthermore, we showed that GST activity can be sensitively assessed with MCB in homogenates of pest insects such as Heliothis virescens, Spodoptera frugiperda (Lepidoptera: Noctuidae), Plutella xylostella (Lepidoptera: Yponomeutidae), and Myzus persicae (Hemiptera: Aphididae). PMID:11950219

  18. Mohorovičić Discontinuity at the Contact of the Adriatic Microplate and Pannonian Segment

    NASA Astrophysics Data System (ADS)

    Sumanovac, F.; Oreskovic, J.

    2009-12-01

    Two-dimensional seismic and gravity modellings were applied to the contact of the Dinarides and the Pannonian basin as a relation between the Adriatic microplate and Pannonian segment, as a contribution to solving structural and tectonic relations. The basic exploration was carried out on the profile Alp07, which was a part of the ALP 2002 experiment (Brückl et al., 2003). This profile stretches in Croatia from Istra to the Drava River at Hungarian-Croatian border in a WSW-ENE direction. Two-dimensional seismic modelling was carried out using tomographic inversion and ray tracing technique. The Moho depth is the greatest in the area of the Dinarides, reaching about 40 km, and is shallowest (30 to 20 km) in the Pannonian basin. In order to obtain additional constraints on the crustal structure, 2 D gravity modelling was also performed. The layer boundaries were retained from the seismic model and varying of densities in the model produced a good fit to the data. A geological model was constructed based on both geophysical models (Šumanovac et al., 2009). Three types of the crust were found along the profile: the Dinaridic and the Pannonian crusts that are separated by a relatively wide Transition zone. The Dinaridic upper crust is characterised by low seismic velocities and densities, but its lower crust has high velocities and densities. The Pannonian crust can be seen as unique layer characterised by both low seismic velocities and densities. Large lateral and vertical changes in densities and seismic velocities can be found in the Transition zone. Gravity modelling on the Alp07 profile enabled a definition of calibrated rock densities that were applied on other profiles set up in the study area, generally covering the area of Croatia and Bosnia-Herzegovina, for the purpose of determining the structural relations. This significantly improved the resolution of gravity modelling, as well. The structural units determined on the Alp07 profile (Pannonian crust

  19. 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

  20. 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. PMID:20388293

  1. 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.

  2. Transdermal film-loaded finasteride microplates to enhance drug skin permeation: Two-step optimization study.

    PubMed

    Ahmed, Tarek A; El-Say, Khalid M

    2016-06-10

    The goal was to develop an optimized transdermal finasteride (FNS) film loaded with drug microplates (MIC), utilizing two-step optimization, to decrease the dosing schedule and inconsistency in gastrointestinal absorption. First; 3-level factorial design was implemented to prepare optimized FNS-MIC of minimum particle size. Second; Box-Behnken design matrix was used to develop optimized transdermal FNS-MIC film. Interaction among MIC components was studied using physicochemical characterization tools. Film components namely; hydroxypropyl methyl cellulose (X1), dimethyl sulfoxide (X2) and propylene glycol (X3) were optimized for their effects on the film thickness (Y1) and elongation percent (Y2), and for FNS steady state flux (Y3), permeability coefficient (Y4), and diffusion coefficient (Y5) following ex-vivo permeation through the rat skin. Morphological study of the optimized MIC and transdermal film was also investigated. Results revealed that stabilizer concentration and anti-solvent percent were significantly affecting MIC formulation. Optimized FNS-MIC of particle size 0.93μm was successfully prepared in which there was no interaction observed among their components. An enhancement in the aqueous solubility of FNS-MIC by more than 23% was achieved. All the studied variables, most of their interaction and quadratic effects were significantly affecting the studied variables (Y1-Y5). Morphological observation illustrated non-spherical, short rods, flakes like small plates that were homogeneously distributed in the optimized transdermal film. Ex-vivo study showed enhanced FNS permeation from film loaded MIC when compared to that contains pure drug. So, MIC is a successful technique to enhance aqueous solubility and skin permeation of poor water soluble drug especially when loaded into transdermal films. PMID:26993962

  3. Reading between the Lines: Societal Norms in Sierra Leonean Readers.

    ERIC Educational Resources Information Center

    Brams, Patricia

    1980-01-01

    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. (Editor)

  4. Readers Use Black Newspapers for Health/Cancer Information

    PubMed Central

    Len-Ríos, María E.; Cohen, Elisia; Caburnay, Charlene

    2010-01-01

    A national survey of readers of black newspapers shows that whether readers depend on black newspapers for cancer and health information depends on their black newspaper use, black self-identity and general media dependency. PMID:21833156

  5. 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.

  6. Child readers' eye movements in reading Thai.

    PubMed

    Kasisopa, Benjawan; Reilly, Ronan G; Luksaneeyanawin, Sudaporn; Burnham, Denis

    2016-06-01

    It has recently been found that adult native readers of Thai, an alphabetic scriptio continua language, engage similar oculomotor patterns as readers of languages written with spaces between words; despite the lack of inter-word spaces, first and last characters of a word appear to guide optimal placement of Thai readers' eye movements, just to the left of word-centre. The issue addressed by the research described here is whether eye movements of Thai children also show these oculomotor patterns. Here the effect of first and last character frequency and word frequency on the eye movements of 18 Thai children when silently reading normal unspaced and spaced text was investigated. Linear mixed-effects model analyses of viewing time measures (first fixation duration, single fixation duration, and gaze duration) and of landing site location revealed that Thai children's eye movement patterns were similar to their adult counterparts. Both first character frequency and word frequency played important roles in Thai children's landing sites; children tended to land their eyes further into words, close to the word centre, if the word began with higher frequency first characters, and this effect was facilitated in higher frequency words. Spacing also facilitated more effective use of first character frequency and it also assisted in decreasing children's viewing time. The use of last-character frequency appeared to be a later development, affecting mainly single fixation duration and gaze duration. In general, Thai children use the same oculomotor control mechanisms in reading spaced and unspaced texts as Thai adults, who in turn have similar oculomotor control as readers of spaced texts. Thus, it appears that eye movements in reading converge on the optimal landing site using whatever cues are available to guide such placement. PMID:27137836

  7. 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).

  8. 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

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

    PubMed Central

    Vijayakumar, Paul Priyesh; Muriana, Peter M.

    2015-01-01

    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. PMID:26111195

  10. 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...

  11. 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...

  12. 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...

  13. 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...

  14. 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...

  15. 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.

  16. 21 CFR 886.5800 - Ophthalmic bar reader.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5800 Ophthalmic bar reader. (a) Identification. An ophthalmic bar reader is a device that consists of a magnifying lens intended for use by a... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Ophthalmic bar reader. 886.5800 Section...

  17. 21 CFR 886.5800 - Ophthalmic bar reader.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5800 Ophthalmic bar reader. (a) Identification. An ophthalmic bar reader is a device that consists of a magnifying lens intended for use by a... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Ophthalmic bar reader. 886.5800 Section...

  18. 21 CFR 886.5800 - Ophthalmic bar reader.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5800 Ophthalmic bar reader. (a) Identification. An ophthalmic bar reader is a device that consists of a magnifying lens intended for use by a... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Ophthalmic bar reader. 886.5800 Section...

  19. 21 CFR 886.5800 - Ophthalmic bar reader.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5800 Ophthalmic bar reader. (a) Identification. An ophthalmic bar reader is a device that consists of a magnifying lens intended for use by a... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Ophthalmic bar reader. 886.5800 Section...

  20. 21 CFR 886.5800 - Ophthalmic bar reader.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5800 Ophthalmic bar reader. (a) Identification. An ophthalmic bar reader is a device that consists of a magnifying lens intended for use by a... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Ophthalmic bar reader. 886.5800 Section...

  1. What's New in Children's Literature? Engaging Readers through Series Books

    ERIC Educational Resources Information Center

    Ward, Barbara A.; Young, Terrell A.

    2007-01-01

    Although experts may debate the literary value of series books and bemoan readers' reluctance to move away from familiar characters and settings, the fact remains that they often draw reluctant readers to the printed word. There is no evidence that reading series books ruins the literary health or moral fiber of readers (Tunnell & Jacobs, 2008).…

  2. Are E-Readers Viable Instructional Delivery Systems?

    ERIC Educational Resources Information Center

    Schcolnik, Miriam

    2002-01-01

    Reports on a study of e-readers, or electronic book readers, that investigated strategies adult users applied to reading in the new medium, kinds of texts users read, and text characteristics for e-reading. Discusses the process of reading, purposes of reading, and whether e-readers are viable instructional delivery systems. (Contains 63…

  3. Science in American School Readers of the Nineteenth Century.

    ERIC Educational Resources Information Center

    Rillero, Peter; Rudolph, Emanuel D.

    In 19th century America the textbook for reading, or "reader," was predominant as the learning tool for young children. Science selections in these readers introduced students to their first formal science instruction. This paper presents an analysis of the science used in 19th century popular readers. Through a synthesis of expert opinion, the…

  4. Reading and Spelling Error Analysis of Native Arabic Dyslexic Readers

    ERIC Educational Resources Information Center

    Abu-rabia, Salim; Taha, Haitham

    2004-01-01

    This study was an investigation of reading and spelling errors of dyslexic Arabic readers ("n"=20) compared with two groups of normal readers: a young readers group, matched with the dyslexics by reading level ("n"=20) and an age-matched group ("n"=20). They were tested on reading and spelling of texts, isolated words and pseudowords. Two…

  5. Graded Readers: How the Publishers Make the Grade

    ERIC Educational Resources Information Center

    Claridge, Gillian

    2012-01-01

    Publishing graded readers is big business, but there is evidence that the texts themselves are not being read in sufficient quantity to improve language proficiency. This article reports on a study of graded readers, focusing on interviews with some major publishers of graded readers, to investigate their production rationales. The findings…

  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. 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…

  8. Reader Use and Understanding of Newspaper Informational Graphics.

    ERIC Educational Resources Information Center

    Pasternack, Steve; Utt, Sandra H.

    While the use of informational graphics in newspapers is increasing, little is known regarding how well readers can understand them and how readers use them. A study investigated whether readers of newspapers read graphics before or after they read the headline/text, and whether people decide to read a graphic device for appearance-related or…

  9. "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…

  10. Retrospective Miscue Analysis with Proficient Adult ESL Readers

    ERIC Educational Resources Information Center

    Wurr, Adrian J.; Theurer, Joan L.; Kim, Koomi J.

    2008-01-01

    This article reports recent research investigating the use of retrospective miscue analysis (RMA) as an instructional strategy with proficient second-language (L2) readers. RMA aims to heighten a reader's awareness of the reading process by involving readers in detailed analysis of their oral reading behavior. Using a cross-case analysis of three…

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

    PubMed

    Wagschal, Kurt; Lee, Charles C

    2012-04-01

    We present here a whole-cell and permeabilized E. coli cell 1° active/inactive microplate screen for β-d-xylosidase, xylanase, endoglucanase, and ferulic acid esterase enzyme activities, which are critical for the enzymatic deconstruction of biomass for fuels and chemicals. Transformants from genomic or mutagenesis-derived libraries are screened using fluorophore-tagged substrate/enzyme activity pairs that are assayed directly in the protein expression host growth media using a minimum of specialized equipment and supplies. PMID:22285853

  12. 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). PMID:24987801

  13. 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

    2015-08-01

    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. PMID:26738081

  14. A microELISA assay for detection of anti-HLA activity of mouse monoclonal antibodies using an Astroscan 2100 automated plate reader.

    PubMed

    Sadler, A M; Krausa, P; Marsh, S G; Heyes, J M; Bodmer, J G

    1992-04-27

    A microenzyme-linked immunosorbent assay (microELISA) method has been developed using an Astroscan 2100 system automated plate reader which was initially designed for tissue typing by a two colour fluorescent microcytotoxicity assay. A 96-well plate ELISA used for screening mouse monoclonal antibodies raised against surface HLA antigens has been modified for use with the Astroscan plate reader and 72-well typing trays. The existing substrate 4-methylumbelliferyl-beta-D-galactoside (4MUG) has been replaced with fluorescein-di-beta-D-galactopyranoside (FDG), to provide a wavelength (530 nm) detectable by the Astroscan or other automated plate readers designed for reading microcytotoxicity assay plates. The assay volumes have also been reduced tenfold for use with Terasaki microtest plates. The assay now has the major advantage of requiring only 5 microliters of test supernatant allowing hybridomas to be screened earlier during a fusion and on a wider cell panel. The use of the large panel which includes B lymphoblastoid cell lines (B-LCL) and mouse L cell transfectants expressing HLA genes, reduces the length of time the hybridomas need to be kept in tissue culture before selection. Other advantages include the reduction in the number of target cells required, smaller volumes of reagents throughout the assay and the ability to screen cytotoxic as well as non-cytotoxic monoclonal antibodies. The sensitivity of this microELISA proved to be comparable with the original assay and so provides an efficient screening method for monoclonal antibodies. PMID:1583310

  15. Analysis of thermoelastic dissipation in circular micro-plate resonators using the generalized thermoelasticity theory of dual-phase-lagging model

    NASA Astrophysics Data System (ADS)

    Guo, F. L.; Song, J.; Wang, G. Q.; Zhou, Y. F.

    2014-05-01

    This study investigates the thermoelastic dissipation of micro-plate resonators by using the generalized thermoelasticity theory of dual-phase-lagging model. Explicit formulae of thermoelastic damping and frequency shift are derived. Influences of the plate thickness and vibration frequency on the thermoelastic damping are examined. Phenomena distinct from those of classical theory are observed in the numerical results of thermoelastic damping in micro-plate resonators. These results may bring new insights into the study of thermoelastic damping at submicrometer or nanometer scale.

  16. 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. PMID:18973283

  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. A comparison of fluorescamine and naphthalene-2,3-dicarboxaldehyde fluorogenic reagents for microplate-based detection of amino acids.

    PubMed

    Bantan-Polak, T; Kassai, M; Grant, K B

    2001-10-15

    The use of appropriate fluorometric derivatization procedures is of considerable importance for accurate determination of amino acids in biological samples and in metal-assisted peptide hydrolysis reactions. It is especially critical for the relative fluorescence intensities (RFI) of equal amounts of amino acids to be as similar as possible. While fluorescamine and naphthalene-2,3-dicarboxaldehyde (NDA) have proven to be excellent fluorogenic reagents for amino acid detection, the effects of various factors such as organic solvent, buffer, and pH have never been rigorously evaluated with respect to normalizing the relative fluorescence intensities of individual amino acids. To this end, here we describe optimized fluorescamine and NDA derivatization reactions that enhance the accuracy of microplate-based detection of amino acids. For both fluorescamine and NDA, we have shown that the RFI values of 16 of 19 amino acids are greater than 70%. Although determination of tryptophan is problematic, this difficulty is overcome by the addition of beta-cyclodextrin to the NDA reaction. In principle, the optimized fluorescamine and NDA microplate procedures reported here can be utilized as complementary techniques for the detection of 19 of 20 naturally occurring amino acids. PMID:11673879

  19. Long-term storage of compound solutions for high-throughput screening by using a novel 1536-well microplate.

    PubMed

    Pfeifer, Martin Joseph; Scheel, Guenther

    2009-06-01

    This report describes the features and the performance of a new and significantly improved 1536-well microplate design. The design allows for simple, automation-friendly, and cost-effective storage of compound solutions for high-throughput screening. The plate design is based on Society for Biomolecular Sciences standards for microplates and can be molded from polystyrene or cycloolefin copolymer, thus making the plate suitable for use with acoustic dispensing as well as other conventional liquid dispensing in the nanoliter range. For a 9:1 DMSO/water mix as solvent, the novel plate design has shown to perform over 4 months with only minor losses in solvent. Thus, this novel plate design creates the basis for further reductions in compound storage volumes and allows for an increase in the storage times for microliter volumes for up to a year or more. The high protection against solvent evaporation is also visible for aqueous solutions, thus allowing for reduced edge effects during screening campaigns. PMID:19487771

  20. 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

  1. 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).

  2. PC-based car license plate reader

    NASA Astrophysics Data System (ADS)

    Hwang, Chung-Mu; Shu, Shyh-Yeong; Chen, Wen-Yu; Chen, Yie-Wern; Wen, Kuang-Pu

    1992-11-01

    A car license plate reader (CLPR) using fuzzy inference and neural network algorithm has been developed in Industrial Technology Research Institute (ITRI) and installed in highway toll stations to identify stolen cars. It takes an average of 0.7 seconds to recognize a car license plate by using a PC with 80486-50 CPU. The recognition rate of the system is about 97%. The techniques of CLPR include vehicle sensing, image grab control, optic pre- processing, lighting, and optic character recognition (OCR). The CLPR can be used in vehicle flow statistics, the checking of stolen vehicles, automatic charging systems in parking lots or garage management, and so on.

  3. Does reader visual fatigue impact interpretation accuracy?

    NASA Astrophysics Data System (ADS)

    Krupinski, Elizabeth A.; Berbaum, Kevin S.

    2010-02-01

    To measure the impact of reader of reader visual fatigue by assessing symptoms, the ability to keep the eye focused on the display and diagnostic accuracy. Twenty radiology residents and 20 radiologists were given a diagnostic performance test containing 60 skeletal radiographic studies, half with fractures, before and after a day of clinical reading. Diagnostic accuracy was measured using area under the proper binormal curve (AUC). Error in visual accommodation was measured before and after each test session and subjects completed the Swedish Occupational Fatigue Inventory (SOFI) and the oculomotor strain subscale of the Simulator Sickness Questionnaire (SSQ) before each session. Average AUC was 0.89 for before work test and 0.85 for the after work test, (F(1,36) = 4.15, p = 0.049 < 0.05). There was significantly greater error in accommodation after the clinical workday (F(1,14829) = 7.81, p = 0.005 < 0.01), and after the reading test (F(1,14829) = 839.33, p < 0.0001). SOFI measures of lack of energy, physical discomfort and sleepiness were higher after a day of clinical reading (p < 0.05). The SSQ measure of oculomotor symptoms (i.e., difficulty focusing, blurred vision) was significantly higher after a day of clinical reading (F(1,75) = 20.38, p < 0.0001). Radiologists are visually fatigued by their clinical reading workday. This reduces their ability to focus on diagnostic images and to accurately interpret them.

  4. Regulating Readers: The Social Origins of the Readers' Advisor in the United States.

    ERIC Educational Resources Information Center

    Luyt, Brendan

    2001-01-01

    The readers' advisory service was a product of social forces operating in the context of early twentieth century capitalism. The work of French regulation theorists provides a framework for analyzing these forces using the concepts of regime of accumulation and mode of regulation. Libraries were connected in a society-wide project, as opposed to…

  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. Research Connections: The San Diego Striving Readers' Project: Building Academic Success for Adolescent Readers

    ERIC Educational Resources Information Center

    McDonald, Trevor; Thornley, Christina; Staley, Rosemary; Moore, David W.

    2009-01-01

    This Research Connections column describes the background to and the research base for the San Diego Unified School Districts' federally funded Striving Readers Project. The curriculum for the project was developed out of a longitudinal study into the literacy experiences of secondary students in New Zealand and from exploratory work in San Diego…

  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. 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…

  9. 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…

  10. Are Avid Readers Lurking in Your Language Arts Classroom? Myths of the Avid Adolescent Reader

    ERIC Educational Resources Information Center

    Wilson, Nance S.; Kelley, Michelle J.

    2010-01-01

    This article describes a pilot study conducted with 10 identified avid adolescent readers who completed the Adolescent Motivation to Read Profile (AMRP) (Pitcher, Albright, DeLaney, Walker, Seunarienesingh, & Moggie, 2007) that includes both a survey to determine students' self-concept and value of reading and an interview that sheds light on what…

  11. Characteristics of Poor Readers of Korean "Hangul": Auditory, Visual and Phonological Processing

    ERIC Educational Resources Information Center

    Kim, Jeesun; Davis, Chris

    2004-01-01

    The vast majority of work on the processing characteristics of poor readers has been conducted with readers of English. In this article we report on results that outline the processing characteristics of poor readers of the Korean alphabetic-syllabic script, "Hangul." Three groups of readers (10 good readers, 10 poor readers and 9 poor readers…

  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. PMID:25576929

  13. Parietal function in good and poor readers

    PubMed Central

    Laycock, Robin; Crewther, Sheila G; Kiely, Patricia M; Crewther, David P

    2006-01-01

    Background While there are many psychophysical reports of impaired magnocellular pathway function in developmental dyslexia (DD), few have investigated parietal function, the major projection of this pathway, in good and poor readers closely matched for nonverbal intelligence. In view of new feedforward-feedback theories of visual processing, impaired magnocellular function raises the question of whether all visually-driven functions or only those associated with parietal cortex functions are equally impaired and if so, whether parietal performance is more closely related to general ability levels than reading ability. Methods Reading accuracy and performance on psychophysical tasks purported to selectively activate parietal cortex such as motion sensitivity, attentional tracking, and spatial localization was compared in 17 children with DD, 16 younger reading-age matched (RA) control children, and 46 good readers of similar chronological-age (CA) divided into CA-HighIQ and a CA-LowIQ matched to DD group nonverbal IQ. Results In the age-matched groups no significant differences were found between DD and CA controls on any of the tasks relating to parietal function, although performance of the DD group and their nonverbal IQ scores was always lower. As expected, CA and RA group comparisons indicated purported parietal functioning improves with age. No difference in performance was seen on any of the parietally driven tasks between the DD and age-nonverbal IQ matched groups, whereas performance differentiated the DD group from the age-matched, higher nonverbal IQ group on several such tasks. An unexpected statistical difference in performance between lower reading age (DD and RA children) and all higher reading age (CA) children was seen on a test of chromatic sensitivity, whereas when high and low nonverbal IQ normal readers were compared performance was not different Conclusion The results indicate that performance on purported parietal functions improves with age

  14. 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

  15. Metabolic effects of azoxystrobin and kresoxim-methyl against Fusarium kyushuense examined using the Biolog FF MicroPlate.

    PubMed

    Wang, Hancheng; Wang, Jin; Chen, Qingyuan; Wang, Maosheng; Hsiang, Tom; Shang, Shenghua; Yu, Zhihe

    2016-06-01

    Azoxystrobin and kresoxim-methyl are strobilurin fungicides, and are effective in controlling many plant diseases, including Fusarium wilt. The mode of action of this kind of chemical is inhibition of respiration. This research investigated the sensitivities of Fusarium kyushuense to azoxystrobin and kresoxim-methyl, and to the alternative oxidase inhibitor salicylhydroxamic acid (SHAM). The Biolog FF MicroPlate is designed to examine substrate utilization and metabolic profiling of micro-organisms, and was used here to study the activity of azoxystrobin, kresoxim-methyl and SHAM against F. kyushuense. Results presented that azoxystrobin and kresoxim-methyl strongly inhibited conidial germination and mycelial growth of F. kyushuense, with EC50 values of 1.60 and 1.79μgml(-1), and 6.25 and 11.43μgml(-1), respectively; while not for SHAM. In the absence of fungicide, F. kyushuense was able to metabolize 91.6% of the tested carbon substrates, including 69 effectively and 18 moderately. SHAM did not inhibit carbon substrate utilization. Under the selective pressure of azoxystrobin and kresoxim-methyl during mycelial growth (up to 100μgml(-1)) and conidial germination (up to 10μgml(-1)), F. kyushuense was unable to metabolize many substrates in the Biolog FF MicroPlate; while especially for carbon substrates in glycolysis and tricarboxylic acid cycle, with notable exceptions such as β-hydroxybutyric acid, y-hydroxybutyric acid, α-ketoglutaric acid, α-d-glucose-1-phosphate, d-saccharic acid and succinic acid in the mycelial growth stage, and β-hydroxybutyric acid, y-hydroxybutyric acid, α-ketoglutaric acid, tween-80, arbutin, dextrin, glycerol and glycogen in the conidial germination stage. This is a new finding for some effect of azoxystrobin and kresoxim-methyl on carbon substrate utilization related to glycolysis and tricarboxylic acid cycle and other carbons, and may lead to future applications of Biolog FF MicroPlate for metabolic effects of other

  16. Chinese Deaf Readers Have Early Access to Parafoveal Semantics

    PubMed Central

    Yan, Ming; Pan, Jinger; Bélanger, Nathalie N.; Shu, Hua

    2014-01-01

    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, parafoveal semantic information is obtained earlier among the deaf readers. In addition, a phonological PB effect was found for the better deaf readers (relative to less-skilled deaf readers) though we also provide an alternative explanation for this effect. Providing evidence that Chinese deaf readers have higher efficiency when processing parafoveal semantics, the study indicates flexibility across individuals in the mechanisms underlying word recognition adapting to the inputs available in the linguistic environment. PMID:24999711

  17. 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

  18. 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.

  19. Improved reader for magnetically-encoded ID cards

    NASA Technical Reports Server (NTRS)

    Wu, T. T.

    1979-01-01

    Hybrid demodulator in electronic card reader for magnetically encoded identification cards, accommodates variations in insertion speeds, yet is simpler and less expensive than equivalent all-digital circuits.

  20. 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. PMID:26549297

  1. Microplate-based screening for small molecule inhibitors of neuropilin-2/vascular endothelial growth factor-C interactions.

    PubMed

    Parker, Matthew W; Vander Kooi, Craig W

    2014-05-15

    Vascular endothelial growth factor-C (VEGF-C) is a secreted growth factor essential for lymphangiogenesis. VEGF-C functions in both physiological and pathological lymphangiogenesis, particularly in tumor metastasis, making it an attractive therapeutic target. Members of two families of cell surface receptors transduce VEGF-C signals: neuropilin-2 (Nrp2) and VEGF-receptor (VEGFR)-2/3. Nrp2 is a promising target for inhibition because it is highly expressed in lymphatic vessels. Here we describe a microplate-based assay for discovery of VEGF-C/Nrp2 inhibitors. We optimize this assay for use in screening an inhibitor library and identify three novel Nrp2/VEGF-C binding inhibitors from the National Institutes of Health (NIH) Clinical Collection small molecule library. PMID:24583243

  2. 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

  3. Large-scale preparation of shape controlled SnO and improved capacitance for supercapacitors: from nanoclusters to square microplates.

    PubMed

    Wang, Lu; Ji, Hongmei; Zhu, Feng; Chen, Zhi; Yang, Yang; Jiang, Xuefan; Pinto, João; Yang, Gang

    2013-08-21

    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. PMID:23842544

  4. A universally calibrated microplate ferric reducing antioxidant power (FRAP) assay for foods and applications to Manuka honey.

    PubMed

    Bolanos de la Torre, Amparo Angelica S; Henderson, Terence; Nigam, Poonam Singh; Owusu-Apenten, Richard K

    2015-05-01

    The ferric reducing antioxidant power (FRAP) assay was recently adapted to a microplate format. However, microplate-based FRAP (mFRAP) assays are affected by sample volume and composition. This work describes a calibration process for mFRAP assays which yields data free of volume effects. From the results, the molar absorptivity (ε) for the mFRAP assay was 141,698 M(-1) cm(-1) for gallic acid, 49,328 M(-1) cm(-1) for ascorbic acid, and 21,606 M(-1) cm(-1) for ammonium ferrous sulphate. The significance of ε (M(-1) cm(-1)) is discussed in relation to mFRAP assay sensitivity, minimum detectable concentration, and the dimensionless FRAP-value. Gallic acid showed 6.6 mol of Fe(2+) equivalents compared to 2.3 mol of Fe(+2) equivalents for ascorbic acid. Application of the mFRAP assay to Manuka honey samples (rated 5+, 10+, 15+, and 18+ Unique Manuka Factor; UMF) showed that FRAP values (0.54-0.76 mmol Fe(2+) per 100g honey) were strongly correlated with UMF ratings (R(2)=0.977) and total phenols content (R(2) = 0.982)whilst the UMF rating was correlated with the total phenols (R(2) = 0.999). In conclusion, mFRAP assay results were successfully standardised to yield data corresponding to 1-cm spectrophotometer which is useful for quality assurance purposes. The antioxidant capacity of Manuka honey was found to be directly related to the UMF rating. PMID:25529660

  5. 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.

  6. 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. PMID:25145941

  7. Alps, Carpathians and Dinarides-Hellenides: about plates, micro-plates and delaminated crustal blocks

    NASA Astrophysics Data System (ADS)

    Schmid, Stefan

    2014-05-01

    Before the onset of Europe-Africa continental collision in the Dinarides-Hellenides (around 60Ma) and in the Alps and Western Carpathians (around 35 Ma), and at a large scale, the dynamics of orogenic processes in the Mediterranean Alpine chains were governed by Europe-Africa plate convergence leading to the disappearance of large parts of intervening oceanic lithosphere, i.e. the northern branch of Neotethys along the Sava-Izmir-Ankara suture and Alpine Tethys along the Valais-Magura suture (Schmid et al. 2008). In spite of this, two major problems concerning the pre-collisional stage are still poorly understood: (1) by now we only start to understand geometry, kinematics and dynamics of the along-strike changes in the polarity of subduction between Alps-Carpathians and Dinarides-Hellenides, and (2) it is not clear yet during exactly which episodes and to what extent intervening rifted continental fragments such as, for example, Iberia-Briançonnais, Tisza, Dacia, Adria-Taurides moved independently as micro-plates, and during which episodes they remained firmly attached to Europa or Africa from which they broke away. As Europe-Africa plate convergence slowed down well below 1 cm/yr at around 30 Ma ago these pre-collisional processes driven by plate convergence on a global scale gave way to more local processes of combined roll-back and crustal delamination in the Pannonian basin of the Carpathian embayment and in the Aegean (as well as in the Western Mediterranean, not discussed in this contribution). In the case of the Carpathian embayment E-directed roll back totally unrelated to Europe-Africa N-S-directed convergence, started at around 20 Ma ago, due to the presence relict oceanic lithosphere in the future Pannonian basin that remained un-subducted during collision. Due to total delamination of the crust from the eastward rolling back European mantle lithosphere the anticlockwise rotating ALCAPA crustal block, consisting of Eastern Alps and Western Carpathian

  8. Are You Ready for E-Readers in Your Library?

    ERIC Educational Resources Information Center

    Gustafson, Chris

    2012-01-01

    This article discusses some things to consider when thinking about adding e-readers to one's collection. These include: (1) What is the purpose?; (2) Which device?; (3) How will one keep track of his or her e-readers and their contents?; (4) What are some potential pitfalls?; (5) Is there a positive impact on student learning?; and (6) Are…

  9. Understanding the Motivation of Deaf Adolescent Latino Struggling Readers

    ERIC Educational Resources Information Center

    Herzig, Melissa Pia

    2009-01-01

    Our methods for educating Deaf adolescent Latino struggling readers need to change in order to maximize their learning. As with all students, this begins with identifying student strengths and building on these strengths to help students gain new and productive skills. We need to find out what motivates these Latino Deaf readers and what will…

  10. 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…

  11. Temporal and Causal Reasoning in Deaf and Hearing Novice Readers

    ERIC Educational Resources Information Center

    Sullivan, Susan; Oakhill, Jane; Arfé, Barbara; Boureux, Magali

    2014-01-01

    Temporal and causal information in text are crucial in helping the reader form a coherent representation of a narrative. Deaf novice readers are generally poor at processing linguistic markers of causal/temporal information (i.e., connectives), but what is unclear is whether this is indicative of a more general deficit in reasoning about…

  12. In Days Gone By: A Reader. Ukrainian Language Development Series.

    ERIC Educational Resources Information Center

    Alberta Dept. of Education, Edmonton.

    This reader contains a selection of short texts and is intended for intermediate- to advanced-level students of Ukrainian. The reader is one of four in the Ukrainian Language Development series; the other three are "Conflict,""Deeds and Doers," and "Decisions." The book includes a variety of poems by Ukrainian national poet Taras Shevchenko, and…

  13. 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…

  14. Author's Intentions and Readers' Interpretations. Technical Report No. 276.

    ERIC Educational Resources Information Center

    Tierney, Robert J.; And Others

    Three studies investigated the nature of the author-reader relationship during discourse comprehension and production, as well as the influence of selected factors on the author-reader relationship. The first study produced data indicating that the relationship is susceptible to subtle variations in the identity of the author. In those situations…

  15. Cutlines/Captions Must Give Readers Meaningful Information.

    ERIC Educational Resources Information Center

    Konkle, Bruce E.

    2002-01-01

    Proposes that scholastic journalists should not shortchange the importance of words under, beside, or below photographs. Notes that in-depth cutlines and captions for scholastic publications and photographs can help hook the reader into the feature on the page; extend the visual coverage of a photograph's content; and give readers specific…

  16. Intervention All Day Long: New Hope for Struggling Readers

    ERIC Educational Resources Information Center

    Allington, Richard L.

    2007-01-01

    School districts have come to think of intervention for struggling readers as something accomplished in a session outside the classroom, one period long, taught by someone other than that student's usual teacher. This often leaves struggling readers in a learning environment where no theory or empirical evidence would predict substantial learning.…

  17. Mosaic of Thought: Teaching Comprehension in a Reader's Workshop.

    ERIC Educational Resources Information Center

    Keene, Ellin Oliver; Zimmermann, Susan

    This book explores the question of how students become thoughtful, independent readers who deeply understand what they read by examining the thought processes of proficient readers. The book then uses these processes as models for the strategies it offers--strategies intended to help children become more flexible, adaptive, independent, and…

  18. Signaling in Text and Its Interaction with Reader Strategies.

    ERIC Educational Resources Information Center

    Meyer, Bonnie J. F.

    Research has identified three reading strategies that are affected in various ways by signaling--the emphasis of superordinate relationships in text. Using a structure strategy, readers follow the text's superordinate relations to focus on the text's message and how it relates to supportive details. Readers who use the detail/list strategy focus…

  19. Using Collaborative Writing Creatively To Teach Reader-Based Prose.

    ERIC Educational Resources Information Center

    Jones, Nathan B.

    This paper describes the basic features of a collaborative writing exercise used to help 40 English-as-a-Foreign-Language (EFL) students write reader-based prose. It presents an actual study that examined the use of collaborative writing to help students draft reader-based prose in intermediate and advanced EFL composition classes at the…

  20. 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…

  1. Semantic Priming Effects in Normal versus Poor Readers

    ERIC Educational Resources Information Center

    Assink, Egbert M. H.; Van Bergen, Floor; Van Teeseling, Heleen; Knuijt, Paul P. N. A.

    2004-01-01

    The authors studied sensitivity to semantic priming, as distinct from semantic judgment, in poor readers. Association strength (high vs. low semantic association) was manipulated factorially with semantic association type (categoric vs. thematic association). Participants were 11-year-old poor readers (n = 15) who were matched with a group of…

  2. Reader's Theatre: The "Theatrical" Approach to Teaching Literature.

    ERIC Educational Resources Information Center

    Ratliff, Gerald Lee

    This document presents a rationale for using reader's theatre in the classroom. It briefly discusses some of the conventions that distinguish reader's theatre from traditional theatrical productions, including direction of voice and action to the audience, and the use of scripts. The major portion of the paper offers seven exercises on the…

  3. 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…

  4. 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…

  5. 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…

  6. 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.)

  7. 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…

  8. 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.…

  9. Guiding Readers to New Understandings through Electronic Text.

    ERIC Educational Resources Information Center

    Patterson, Nancy, Ed.; Pipkin, Gloria, Ed.

    2001-01-01

    Argues that computer technology can help to engage struggling readers in meaningful transactions with text. Lists and describes seven web sites that will captivate reluctant readers. Notes three web sites that send students on "WebQuests" to transact with text in order to build knowledge. Discusses other ways to engage students in text via…

  10. 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…

  11. First Encounters: Books to Encourage the Beginning Reader.

    ERIC Educational Resources Information Center

    Landsberg, Michele

    1992-01-01

    Describes over 20 picture books that motivate beginning readers to become more proficient at reading and to develop a love of reading. Discusses the role of illustrations in engaging the reader and contributing to story development. Recommends parental assistance to help children enjoy the books' humor and subtitles. (JB)

  12. Readers as Writers Composing from Sources. Technical Report No. 18.

    ERIC Educational Resources Information Center

    Spivey, Nancy Nelson; King, James R.

    A study of discourse synthesis (readers/writers composing new texts by selecting, organizing, and connecting content from source texts) examined the performance of accomplished and less accomplished readers in the 6th, 8th, and 10th grades on a report-writing task. Over a 3-day period 60 English/language arts students wrote informational reports…

  13. 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.…

  14. 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,…

  15. William S. Gray and the Dick and Jane Readers.

    ERIC Educational Resources Information Center

    Mavrogenes, Nancy A.

    William S. Gray, Professor of Education and Dean of the College of Education at the University of Chicago, began work in 1929 on a major revision of the "Elson Readers," a popular basal series published by Scott, Foresman and Company. Organized around the daily life of two ordinary children, the "Dick and Jane" readers ultimately became the most…

  16. Reading Strategies Employed by Greek Braille Readers: Miscue Analysis

    ERIC Educational Resources Information Center

    Vakali, Anthi; Evans, Roy

    2007-01-01

    Reading development is said to progress in a series of developmental skills and stages that need to be acquired by emerging young readers. The literature on Greek braille reading and its development found that print readers opt for the same text decoding strategy even at the initial stages of development. This article explores the development of…

  17. 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…

  18. Implementing Readers Theatre as an Approach to Classroom Fluency Instruction

    ERIC Educational Resources Information Center

    Young, Chase; Rasinski, Timothy

    2009-01-01

    This article describes how Readers Theatre was implemented as an engaging means for reading fluency instruction in a second-grade classroom. A weekly Readers Theatre format was utilized over the course of a school year. The format was added to the existing literacy program. Data collected during the school year revealed significant gains in…

  19. 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…

  20. 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…

  1. 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)

  2. Be That Teacher! Breaking the Cycle for Struggling Readers

    ERIC Educational Resources Information Center

    Risko, Victoria J.; Walker-Dalhouse, Doris

    2012-01-01

    Tens of thousands of students begin each new school year with the hope that they will finally find "the" teacher who will help them succeed as readers, writers, and learners. This book shows how teachers can provide the type of differentiated instruction that struggling readers need by drawing on students' individual and cultural backgrounds, as…

  3. Envisioning Story: The Eye Movements of Beginning Readers

    ERIC Educational Resources Information Center

    Duckett, Peter

    2003-01-01

    Miscue analysis and eye movement analysis are used to explore the reading process of first-grade beginning readers as they use pictures and print in a picture book designed for instructional purposes. Eye movement miscue analysis (EMMA) is also used as a tool to gain insights into the reading strategies of the beginning readers in this study.…

  4. "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…

  5. 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…

  6. A Longitudinal Literacy Profile of Greek Precocious Readers

    ERIC Educational Resources Information Center

    Tafa, Eufimia; Manolitsis, George

    2008-01-01

    The aim of this five-year longitudinal study was to examine whether 13 Greek precocious readers' performance on a variety of reading, spelling, and phonological-awareness tasks from kindergarten through the fourth grade was different from that of 12 Greek nonprecocious readers and, if there were differences between the two groups' performances,…

  7. Let's Not Leave Advanced and Gifted Readers "behind"

    ERIC Educational Resources Information Center

    Weber, Christine

    2010-01-01

    Weber issues a challenge to teachers grappling with the demands (and failures) of NCLB, and asks them to make certain that advanced and gifted readers are not left behind. To that end, she shares characteristics of gifted readers, ways to support them, and tips for teaching that will result in doing "a much better job of providing appropriate…

  8. 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); "Distance…

  9. 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 take a computerized…

  10. Towards More Secure Biometric Readers for Effective Digital Forensic Investigation

    NASA Astrophysics Data System (ADS)

    Trabelsi, Zouheir; Al-Hemairy, Mohamed; Baggili, Ibrahim; Amin, Saad

    This paper investigates the effect of common network attacks on the performance, and security of several biometric readers. Experiments are conducted using Denial of Service attacks (DoSs) and the ARP cache poisoning attack. The experiments show that the tested biometric readers are vulnerable to DoS attacks, and their recognition performance is significantly affected after launching the attacks. However, the experiments show that the tested biometric readers are secure from the ARP cache poisoning attack. This work demonstrates that biometric readers are easy targets for malicious network users, lack basic security mechanisms, and are vulnerable to common attacks. The confidentiality, and integrity of the log files in the biometric readers, could be compromised with such attacks. It then becomes important to study these attacks in order to find flags that could aid in a network forensic investigation of a biometric device.

  11. Materials management with a bar code reader.

    PubMed

    Kaplan, R S

    1990-01-01

    A materials management system capable of inventory control, accounting and the automatic recording of supplies for a clinical department has been developed for the George Washington University Hospital Department of Anesthesia. This system combines a microprocessor-based computer for data storage and a hand-held bar code reader to record the bar code scan of each item in the inventory. A relational software program with easy-to-use menus and help keys was written. Bar code information stored for each item includes item number, quantity, date and time of issue. Accumulated bar code scans are loaded into the computer by use of a serial port and then used to update current inventory in the computer. Comparison between current inventory and reorder levels by the computer will initiate automatic printing of appropriate purchase orders. Reorder levels are adjusted regularly, by comparing previous year or month usage to current needs; items already on order, items on back order and delivery lag time are also taken into account. PMID:10104851

  12. 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.

  13. 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…

  14. A Microplate Format Assay for Real-Time Screening for New Aldolases that Accept Aryl-Substituted Acceptor Substrates.

    PubMed

    Ma, Huan; Enugala, Thilak Reddy; Widersten, Mikael

    2015-12-01

    Aldolases are potentially important biocatalysts for asymmetric synthesis of polyhydroxylated compounds. Fructose 6-phosphate aldolase (FSA) is of particular interest by virtue of its unusually relaxed dependency on phosphorylated substrates. FSA has been reported to be a promising catalyst of aldol addition involving aryl-substituted acceptors such as phenylacetaldehyde that can react with donor ketones such as hydroxyacetone. Improvement of the low intrinsic activity with bulky acceptor substrates of this type is of great interest but has been hampered by the lack of powerful screening protocols applicable in directed evolution strategies. Here we present a new screen allowing for direct spectrophotometric recording of retro-aldol cleavage. The assay utilizes an aldehyde reductase produced in vitro by directed evolution; it reduces the aldehyde product formed after cleavage of the aldol by FSA. The assay is suitable both for steady-state enzyme kinetics and for real-time activity screening in a 96-well format. PMID:26449620

  15. 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.

  16. 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

  17. History Textbooks for Poor Readers--At What Cost?

    ERIC Educational Resources Information Center

    Smith, Allen

    1980-01-01

    Analyzes seven junior high school American history textbooks for slow readers. Results indicate weaknesses in historical explanations, causal-effect relationships, background, and synthesizing historical information, specifically when depicting the Civil War and the Great Depression. (Author/KC)

  18. Editorial Comment: Practice and Inquiry: "EJ" Readers and "EJ" Writers.

    ERIC Educational Resources Information Center

    English Journal, 1988

    1988-01-01

    Points out that the "English Journal" considers all readers prospective contributors. Lists and discusses the criteria that "English Journal" uses to judge manuscripts that purport to describe effective classroom practices. (ARH)

  19. A combination turbidity and supernatant microplate assay to rank-order the supersaturation limits of early drug candidates.

    PubMed

    Morrison, John S; Nophsker, Michelle J; Haskell, Roy J

    2014-10-01

    A unique opportunity exists at the drug discovery stage to overcome inherently poor solubility by selecting drug candidates with superior supersaturation propensity. Existing supersaturation assays compare either precipitation-resistant or precipitation-inhibiting excipients, or higher-energy polymorphic forms, but not multiple compounds or multiple concentrations. Furthermore, these assays lack sufficient throughput and compound conservation necessary for implementation in the discovery environment. A microplate-based combination turbidity and supernatant concentration assay was therefore developed to determine the extent to which different compounds remain in solution as a function of applied concentration in biorelevant media over a specific period of time. Dimethyl sulfoxide stock solutions at multiple concentrations of four poorly soluble, weak base compounds (Dipyridamole, Ketoconazole, Albendazole, and Cinnarizine) were diluted with pH 6.5 buffer as well as FaSSIF. All samples were monitored for precipitation by turbidity at 600 nm over 1 h and the final supernatant concentrations were measured. The maximum supersaturation ratio was calculated from the supersaturation limit and the equilibrium solubility in each media. Compounds were rank-ordered by supersaturation ratio: Ketoconazole > Dipyridamole > Cinnarizine ∼ Albendazole. These in vitro results correlated well with oral AUC ratios from published in vivo pH effect studies, thereby confirming the validity of this approach. PMID:25070886

  20. Rapid, low-technology MIC determination with clinical Mycobacterium tuberculosis isolates by using the microplate Alamar Blue assay.

    PubMed

    Franzblau, S G; Witzig, R S; McLaughlin, J C; Torres, P; Madico, G; Hernandez, A; Degnan, M T; Cook, M B; Quenzer, V K; Ferguson, R M; Gilman, R H

    1998-02-01

    A colorimetric, microplate-based Alamar Blue assay (MABA) method was used to determine the MICs of isoniazid (INH), rifampin, streptomycin (SM), and ethambutol (EMB) for 34 Peruvian Mycobacterium tuberculosis isolates (including both pansensitive and multidrug-resistant strains) and the H37Rv strain by using bacterial suspensions prepared directly from solid media. Results for all isolates were available within 8 days. Discordant results were observed on initial tests for 3 of 16 INH-susceptible isolates, 5 of 31 EMB-susceptible isolates, and 2 of 4 SM-resistant isolates (by the BACTEC 460 system). The overall agreements between the MICs obtained by MABA and the results obtained with the BACTEC 460 system were 87.9% for initial results and 93.6% after retesting 12 of 17 samples with discrepant results. Interpretation of MABA endpoints improved with technical experience. The MABA is a simple, rapid, low-cost, appropriate technology which does not require expensive instrumentation and which makes use of a nontoxic, temperature-stable reagent. PMID:9466742

  1. Rapid, Low-Technology MIC Determination with Clinical Mycobacterium tuberculosis Isolates by Using the Microplate Alamar Blue Assay

    PubMed Central

    Franzblau, Scott G.; Witzig, Richard S.; McLaughlin, James C.; Torres, Patricia; Madico, Guillermo; Hernandez, Antonio; Degnan, Michelle T.; Cook, Mary B.; Quenzer, Virginia K.; Ferguson, Robert M.; Gilman, Robert H.

    1998-01-01

    A colorimetric, microplate-based Alamar Blue assay (MABA) method was used to determine the MICs of isoniazid (INH), rifampin, streptomycin (SM), and ethambutol (EMB) for 34 Peruvian Mycobacterium tuberculosis isolates (including both pansensitive and multidrug-resistant strains) and the H37Rv strain by using bacterial suspensions prepared directly from solid media. Results for all isolates were available within 8 days. Discordant results were observed on initial tests for 3 of 16 INH-susceptible isolates, 5 of 31 EMB-susceptible isolates, and 2 of 4 SM-resistant isolates (by the BACTEC 460 system). The overall agreements between the MICs obtained by MABA and the results obtained with the BACTEC 460 system were 87.9% for initial results and 93.6% after retesting 12 of 17 samples with discrepant results. Interpretation of MABA endpoints improved with technical experience. The MABA is a simple, rapid, low-cost, appropriate technology which does not require expensive instrumentation and which makes use of a nontoxic, temperature-stable reagent. PMID:9466742

  2. Reduced turn-around time for Mycobacterium tuberculosis drug susceptibility testing with a proportional agar microplate assay.

    PubMed

    Nguyen, V A T; Nguyen, H Q; Vu, T T; Nguyen, N A T; Duong, C M; Tran, T H T; Nguyen, H V; Dang, D A; Bañuls, A-L

    2015-12-01

    Multidrug-resistant tuberculosis is a major issue worldwide; however, accessibility to drug susceptibility testing (DST) is still limited in developing countries, owing to high costs and complexity. We developed a proportion method on 12-well microplates for DST. The assay reduced the time to results to <12 days and <10 days when bacterial growth was checked with the naked eye or a microscope, respectively. Comparison with the Canetti-Grosset method showed that the results of the two assays almost overlapped (kappa index 0.98 (95% CI 0.91-1.00) for isoniazid, rifampicin, streptomycin; and kappa index 0.92 (95% CI 0.85-0.99) for ethambutol). The sequencing of genes involved in drug resistance showed similar level of phenotype-genotype agreement between techniques. Finally, measurement of the MICs of rifampicin and ethambutol suggests that the currently used critical ethambutol concentration should be revised, and that the current molecular drug susceptibility tests for rifampicin need to be re-evaluated, as in vitro rifampicin-sensitive isolates could harbour drug resistance-associated mutation(s). PMID:26348263

  3. 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

    2016-05-01

    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.

  4. Silver nanocube on gold microplate as a well-defined and highly active substrate for SERS detection.

    PubMed

    Xia, Xiaohu; Rycenga, Matthew; Qin, Dong; Xia, Younan

    2013-10-14

    Strong enhancement and good reproducibility in Raman signals are two major requirements for a surface-enhanced Raman scattering (SERS) substrate to be used for sensitive detection of an analyte. Here we report a new type of SERS substrate that was fabricated by depositing a Ag nanocube (AgNC) on the surface of a Au microplate (AuMP). Owing to the strong and reproducible hot spots formed at corner sites of the AgNC in proximity with the AuMP surface, the new substrate showed high sensitivity and reproducibility. Using 1,4-benzenedithiol as a probe, the SERS enhancement factor of a typical "AgNC on AuMP" substrate could reach a level as high as 4.7×10(7). In addition to the high sensitivity and reproducibility, the "AgNC on AuMP" substrate also displayed very good stability. Potential use of the "AgNC on AuMP" substrate was demonstrated by detecting crystal violet with high sensitivity. PMID:24187611

  5. Molecular characterization and expression profiles of olfactory receptor genes in the parasitic wasp, Microplitis mediator (Hymenoptera: Braconidae).

    PubMed

    Ma, Long; Gu, Shao-Hua; Liu, Ze-Wen; Wang, Shan-Ning; Guo, Yu-Yuan; Zhou, Jing-Jiang; Zhang, Yong-Jun

    2014-01-01

    Olfactory receptors (OR) are believed to fulfil an indispensable role in insects host-seeking, mating and ovipositing. We obtained 4785 high-quality expressed sequencing tags (EST) from the antennal cDNA library of the parasitic wasp Microplitis mediator, a natural enemy of crop pests. After assembling, 786 contigs and 2130 singletons were generated. Using BLAST searches, a number of olfactory-related genes were identified, including ESTs encoding for 25 ORs. 14 full-length OR genes were cloned and their expression profiles in the wasp olfactory organs were quantified by real-time qRT-PCR. The results indicated a diverse distribution between the tissues and genders, yet the majority of OR genes are highly expressed in antennae. Three OR genes (Or2, Or12 and Or13) are highly expressed in female antennae, eight OR genes (ORco, Or3, Or4, Or5, Or7, Or8, Or9 and Or11) are highly expressed in male antennae. This is the first report on tissue-specific expression of wasp OR genes. Our study provides a foundational knowledge to explore and understand the molecular basis of odorant reception in this parasitic wasp and for the study of trophic interactions of natural enemy. PMID:24291166

  6. 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.

  7. 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.

  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. 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.

  10. Did the Kyrenia Range of northern Cyprus rotate with the Troodos-Hatay microplate during the tectonic evolution of the eastern Mediterranean?

    NASA Astrophysics Data System (ADS)

    Morris, Antony; Robertson, Alastair H. F.; Anderson, Mark W.; Hodgson, Emma

    2016-01-01

    Previous palaeomagnetic studies have allowed the recognition of a distinctive area of Neotethyan oceanic rocks, including the Troodos ophiolite in Cyprus and the Hatay ophiolite to the east in southern Turkey, that underwent 90° of anticlockwise rotation between Late Cretaceous (Campanian) and Early Eocene time. The southern and western boundaries of this rotated Troodos-Hatay microplate have been inferred to lie within, or adjacent to, zones of deformed oceanic and continental margin rocks that are now exposed in southern and western Cyprus; however, the northern boundary of the microplate remains undefined. Relevant to this problem, palaeomagnetic data are presented here from basaltic lavas exposed along the Kyrenia Range, mostly from Late Cretaceous (Maastrichtian) sites and one Eocene site. A positive inclination-only fold test demonstrates that remanences are pre-deformational in age, and positive conglomerate tests show that magnetic remanences were acquired before Late Eocene-Early Oligocene time, together suggesting that primary magnetizations are preserved. Data from the eastern Kyrenia Range and the Karpas Peninsula (the easternmost extension of the Kyrenia Range) document significant relative tectonic rotation between these localities, with no rotation in the eastern range versus 30° of anticlockwise rotation of the Karpas Peninsula. Unfortunately, palaeomagnetic sites from the western Kyrenia Range did not yield tectonically interpretable magnetization directions, probably due to complex poly-phase thrusting and folding, and the central range also yielded no interpretable data. However, the available palaeomagnetic data are sufficient to demonstrate that the Kyrenia terrane underwent a separate rotation history to the Troodos-Hatay microplate and also implies that the northern boundary of the Troodos-Hatay microplate was located between the Troodos ophiolite and the Kyrenia Range. The former microplate margin has since been overridden and concealed by

  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. PMID:27014113

  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. High resolution as a key feature to perform accurate ELISPOT measurements using Zeiss KS ELISPOT readers.

    PubMed

    Malkusch, Wolf

    2005-01-01

    The enzyme-linked immunospot (ELISPOT) assay was originally developed for the detection of individual antibody secreting B-cells. Since then, the method has been improved, and ELISPOT is used for the determination of the production of tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, or various interleukins (IL)-4, IL-5. ELISPOT measurements are performed in 96-well plates with nitrocellulose membranes either visually or by means of image analysis. Image analysis offers various procedures to overcome variable background intensity problems and separate true from false spots. ELISPOT readers offer a complete solution for precise and automatic evaluation of ELISPOT assays. Number, size, and intensity of each single spot can be determined, printed, or saved for further statistical evaluation. Cytokine spots are always round, but because of floating edges with the background, they have a nonsmooth borderline. Resolution is a key feature for a precise detection of ELISPOT. In standard applications shape and edge steepness are essential parameters in addition to size and color for an accurate spot recognition. These parameters need a minimum spot diameter of 6 pixels. Collecting one single image per well with a standard color camera with 750 x 560 pixels will result in a resolution much too low to get all of the spots in a specimen. IFN-gamma spots may have only 25 microm diameters, and TNF-alpha spots just 15 microm. A 750 x 560 pixel image of a 6-mm well has a pixel size of 12 microm, resulting in only 1 or 2 pixel for a spot. Using a precise microscope optic in combination with a high resolution (1300 x 1030 pixel) integrating digital color camera, and at least 2 x 2 images per well will result in a pixel size of 2.5 microm and, as a minimum, 6 pixel diameter per spot. New approaches try to detect two cytokines per cell at the same time (i.e., IFN-gamma and IL-5). Standard staining procedures produce brownish spots (horseradish peroxidase) and blue spots

  14. Nucleopolyhedrovirus infection and/or parasitism by Microplitis pallidipes Szepligeti affect hemocyte apoptosis of Spodoptera exigua (Hübner) larvae.

    PubMed

    Wan, Nian-Feng; Ji, Xiang-Yun; Zhang, Hao; Yang, Jun-Hua; Jiang, Jie-Xian

    2015-11-01

    We determined the effects of parasitism by the endoparasitoid Microplitis pallidipes Szepligeti and/or nucleopolyhedrovirus (NPV) infection on hemocyte apoptosis of Spodoptera exigua (Hübner) larvae. Compared to healthy (control) larvae, larvae that were parasitized, virus-infected, or both all showed a significant increase in hemocyte apoptosis during 48-h observation period. The peaks of hemocyte apoptosis in parasitized, virus-infected and parasitized+infected larvae were at 12, 24 and 48 h after treatment, and were 86.7±1.9, 87.4±3.6 and 76.5±1.6%, respectively. Meanwhile, compared to parasitized larvae, hemocyte apoptosis in jointly parasitized and infected larvae increased by 12.9%, 18.7% and 2.8% at 8, 36 and 48 h respectively, and decreased by 39.0% and 9.1% at 12 and 24h. Compared to virus-infected larvae, hemocyte apoptosis in jointly parasitized and infected larvae increased by 13.4%, 2.4% and 15.3% at 8, 36 and 48 h, respectively, and decreased by 4.0% and 29.9% at 12 and 24h. Our study found that joint and separate parasitism and SeNPV infection induced hemocyte apoptosis of S. exigua larvae. It also revealed that NPV infection promoted host hemocyte apoptosis induced by parasitism at early egg and larval stages of M. pallidipes in host larvae, but inhibited the same effect at late egg stage of M. pallidipes in host larvae, and that parasitism promoted host hemocyte apoptosis induced by NPV infection at early egg and larval stages of M. pallidipes in host larvae, but inhibited the same effect at late egg stage of M. pallidipes in host larvae. PMID:26470677

  15. Micro-plate tectonics and kinematics in Northeast Asia inferred from a dense set of GPS observations

    NASA Astrophysics Data System (ADS)

    Jin, Shuanggen; Park, Pil-Ho; Zhu, Wenyao

    2007-05-01

    The plate tectonics of Northeast Asia are very complex with diffuse and sparse seismicity in the broad plate deformation zones embedded by a number of micro-plates, particularly the controversial Amurian plate. Now the increasingly dense GPS networks in this area provide an important tool to investigate plate tectonic kinematics and to identify the approximate plate tectonic geometries. In this paper, we have processed GPS data (1998-2005) collected by an extensive GPS network (China and South Korea) with more than 85 continuous sites and about 1000 campaign GPS stations. The kinematics of Northeast Asia is studied by modeling GPS-derived velocities with rigid block rotations and elastic deformation. We find that the deformation in Northeast Asia can be well described by a number of rotating blocks, which are independent of the Eurasian plate motion with statistical significance above the 99% confidence level. The tectonic boundary between the North China and Amuria plates is the Yin Shan-Yan Shan Mountain belts with about 2.4 mm/yr extension. Along the boundary between North China and South China, the Qinling-Dabie fault is moving left laterally at about 3.1 mm/yr. The Amuria and South Korea blocks are extending at about 1.8 mm/yr. The Baikal Rift between the Amurian and Eurasian plates is spreading at about 3.0 mm/yr. The 9-17 mm/yr relative motion between the Amuria and Okhotsk blocks is accommodated at the East Sea-Japan trench zone. Localized deformation near the Qinling-Dabie fault and Yin Shan-Yan Shan Mountain belts may be elastic strain accumulation due to interseismic locking of faults.

  16. A Transcriptome Analysis Suggests Apoptosis-Related Signaling Pathways in Hemocytes of Spodoptera litura After Parasitization by Microplitis bicoloratus

    PubMed Central

    Zhang, Yan; Yu, Dongshuai; Yang, Minjun; Yang, Yang; Hu, Jiansheng; Luo, Kaijun

    2014-01-01

    Microplitis bicoloratus parasitism induction of apoptotic DNA fragmentation of host Spodoptera litura hemocytes has been reported. However, how M. bicoloratus parasitism regulates the host signaling pathways to induce DNA fragmentation during apoptosis remains unclear. To address this question, we performed a new RNAseq-based comparative analysis of the hemocytes transcriptomes of non-parasitized and parasitized S. litura. We were able to assemble a total of more than 11.63 Gbp sequence, to yield 20,571 unigenes. At least six main protein families encoded by M. bicoloratus bracovirus are expressed in the parasitized host hemocytes: Ankyrin-repeat, Ben domain, C-type lectin, Egf-like and Mucin-like, protein tyrosine phosphatase. The analysis indicated that during DNA fragmentation and cell death, 299 genes were up-regulated and 2,441 genes were down-regulated. Data on five signaling pathways related with cell death, the gap junctions, Ca2+, PI3K/Akt, NF-κB, ATM/p53 revealed that CypD, which is involved in forming a Permeability Transition Pore Complex (PTPC) to alter mitochondrial membrane permeabilization (MMP), was dramatically up-regulated. The qRT-PCR also provided that the key genes for cell survival were down-regulated under M. bicoloratus parasitism, including those encoding Inx1, Inx2 and Inx3 of the gap junction signaling pathway, p110 subunit of the PI3K/Akt signaling pathway, and the p50 and p65 subunit of the NF-κB signaling pathway. These findings suggest that M. bicoloratus parasitism may regulate host mitochondria to trigger internucleosomal DNA fragmentation. This study will facilitate the identification of immunosuppression-related genes and also improves our understanding of molecular mechanisms underlying polydnavirus-parasitoid-host interaction. PMID:25350281

  17. 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.

  18. 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

  19. 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

  20. Engaging Literacy: A Case Study in the Use of e-Reader Devices

    ERIC Educational Resources Information Center

    Lettenmaier, Kris

    2013-01-01

    The purpose of this case study was to determine to what extent e-readers affected struggling readers as a part of reading instruction at the middle school level to improve students' literacy skills of comprehension, fluency, and vocabulary, and to show how e-readers affected engagement and self-efficacy of struggling readers to read and use…