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Sample records for 96-well microplate format

  1. 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

  2. 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

  3. 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.

  4. 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

  5. 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

  6. 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}.

  7. 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

  8. 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...

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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.

  14. 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

  15. 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

  16. 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

  17. 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.

  18. 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...

  19. 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...

  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. 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

  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. 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

  4. 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

  5. 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.

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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

  11. 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

  12. 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…

  13. 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

  14. 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.

  15. 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…

  16. 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

  17. 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

  18. 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

  19. 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

  20. Direct comparison of the Ames microplate format (MPF) test in liquid medium with the standard Ames pre-incubation assay on agar plates by use of equivocal to weakly positive test compounds.

    PubMed

    Flückiger-Isler, Sini; Kamber, Markus

    2012-08-30

    The Ames microplate format (MPF™) test, which uses liquid media and in 384-well microplates with a readout based on a colour-change, has been used for over 10 years at several major pharmaceutical companies for screening the genotoxic potential of early drug candidates when compound supply is minimal. Meanwhile, Xenometrix has adapted this screen from the two-strain Ames II test for use with five tester strains, in compliance with OECD Guideline 471. A set of 15 equivocal to weakly positive chemicals selected from the National Toxicology Program (NTP) database was tested simultaneously in the Ames microplate format (MPF) and the standard Ames pre-incubation method on agar plates. Such a direct comparison of the two test methods with the same overnight culture(s), chemicals and S9-mix preparation should exclude external variability factors. Thirteen of the 15 chemicals showed concordant results in both tests despite the choice of chemicals that showed varying inter- and even intra-laboratory results in the NTP studies. These results indicate that the Ames MPF™ assay is a reliable predictive tool that can be used like the regular Ames test to evaluate compounds for mutagenicity. PMID:22579797

  1. 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

  2. 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

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

    EPA Science Inventory

    One of the approaches for reducing uncertainties in the assessment of human exposure is to better characterize concentrations of hazardous compounds that may be present in our immediate environment. A significant limitation to this approach, however, is that sampling and labora...

  4. 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

  5. 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...

  6. 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.

  7. 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.

  8. 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

  9. 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.

  10. 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

  11. 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

  12. 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

  13. 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

  14. 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

  15. 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

  16. 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

  17. 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

  18. 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.

  19. 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

  20. 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

  1. IDENTIFICATION OF SALMONELLA-POSITIVE FECAL SAMPLES USING A 96-WELL MICROCULTURE PLATE TECHNIQUE (RX METHOD)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Conventional Salmonella isolation involves multiple sample transfers to culture media performed by an experienced microbiologist. The modified semi-solid RV and XLT (RX) Plate method, a modification of the RX tube format designed by Gailey et al. (2004), consolidates pre-enrichment (buffered pepton...

  2. SIMPLE DEVICE TO DELIVER BEADS TO 96-WELL PLATES FOR RAPID RESUSPENSION OF CELLS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genomic sequencing involves sequence determination of inserts from a large numbers of plasmids of a genomic library. In high throughput DNA sequencing projects, cultures are grown in a 96-deep well plate format, centrifuged, and resuspended. Resuspension of cells was aided by use of a simple devic...

  3. 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

  4. 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.

  5. 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

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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

  14. 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

  15. 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

  16. 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

  17. 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

  18. 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

  19. 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

  20. 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

  1. 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

  2. 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

  3. 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.

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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.

  9. 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

  10. 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

  11. 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

  12. 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.

  13. 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

  14. 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

  15. 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

  16. 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

  17. 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

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

    NASA Astrophysics Data System (ADS)

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

    1991-06-01

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

  19. An antibody microarray, in multiwell plate format, for multiplex screening of foodborne pathogenic bacteria and biomolecules

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Intoxication and infection caused by foodborne pathogens are important problems in the United States, and screening tests for multiple pathogen detection have been developed because food producers are known reservoirs of multiple pathogens. We developed a 96-well microplate, multiplex antibody micr...

  20. 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.

  1. 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

  2. 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.

  3. 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

  4. 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

  5. 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

  6. 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

  7. 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

  8. Wort free amino nitrogen analysis adapted to a microplate format

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The standard method for determining wort free amino nitrogen content calls for the use of test tubes and glass marbles, as well as boiling and 20°C water baths. In this paper we describe how the standard method can be updated and streamlined by replacing water baths, test tubes and marbles with a th...

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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

  14. 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%.

  15. 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

  16. 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

  17. 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

  18. 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

  19. 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.

  20. 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

  1. 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

  2. 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.

  3. 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

  4. 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.

  5. A novel colorimetric assay of β-D-glucans in basidiomycete strains by alcian blue dye in a 96-well microtiter plate.

    PubMed

    C Semedo, Magda; Karmali, Amin; Fonseca, Luís

    2015-01-01

    Basidiomycete strains synthesize several types of β-d-glucans, which play a major role in the medicinal properties of mushrooms. Therefore, the specific quantification of these β-d-glucans in mushroom strains is of great biochemical importance. Because published assay methods for these β-d-glucans present some disadvantages, a novel colorimetric assay method for β-d-glucan with alcian blue dye was developed. The complex formation was detected by following the decrease in absorbance in the range of 620 nm and by hypsochromic shift from 620 to 606 nm (∼14 nm) in UV-Vis spectrophotometer. Analysis of variance was used for optimization of the slope of the calibration curve by using the assay mixture containing 0.017% (w/v) alcian blue in 2% (v/v) acetic acid at pH 3.0. The high-throughput colorimetric assay method on microtiter plates was used for quantification of β-d-glucans in the range of 0-0.8 μg, with a slope of 44.15 × 10(-2) and a limit of detection of 0.017 μg/well. Recovery experiments were carried out by using a sample of Hericium erinaceus, which exhibited a recovery of 95.8% for β-1,3-d-glucan. The present assay method exhibited a 10-fold higher sensitivity and a 59-fold lower limit of detection compared with the published method with congo red. β-d-glucans of several mushrooms strains were isolated from fruiting bodies and mycelia, and they were quantified by this assay method. This assay method is fast, specific, simple, and it can be used to quantify β-d-glucans from other biological sources. PMID:26317582

  6. 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

  7. Released exopolysaccharide (r-EPS) produced from probiotic bacteria reduce biofilm formation of enterohemorrhagic Escherichia coli O157:H7.

    PubMed

    Kim, Younghoon; Oh, Sejong; Kim, Sae Hun

    2009-02-01

    Here, we characterized released-exopolysaccharides (r-EPS) from Lactobacillus acidophilus A4 with the goal of identifying natural compounds that represses biofilm formation. In plastic 96-well microplates that contained 1.0 mg/ml of r-EPS, enterohemorrhagic Escherichia coli (EHEC) biofilms were dramatically decreased by 87% and 94% on polystyrene and polyvinyl chloride (PVC) surfaces, respectively. In the presence of r-EPS, neither their growth rate nor their autoinducer-2-like activity was affected on the EHEC O157:H7. Importantly, consistent reduction in biofilm formation was also observed when r-EPS was applied to the continuous-flow chamber models. In addition, we found that adding r-EPS significantly repressed biofilm formation by affecting genes related to curli production (crl, csgA, and csgB) and chemotaxis (cheY) in transcriptome analysis. Furthermore, these r-EPS could prevent biofilm formation by a wide range of Gram-negative and -positive pathogens. This property may lead to the development of novel food-grade adjuncts for microbial biofilm control. PMID:19103165

  8. 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

  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. 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

  11. 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

  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. 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.

  14. 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

  15. 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

  16. 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

  17. 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

  18. 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.

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

    PubMed

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

    2014-10-01

    Microalgae cultivation conditions in microplates will differ from large-scale photobioreactors in crucial parameters such as light profile, mixing and gas transfer. Hence volumetric productivity (P(v)) measurements made in microplates cannot be directly scaled up. Here we demonstrate that it is possible to use microplates to measure characteristic exponential growth rates and determine the specific growth rate light intensity dependency (μ-I curve), which is useful as the key input for several models that predict P(v). Nannochloropsis salina and Chlorella sorokiniana specific growth rates were measured by repeated batch culture in microplates supplied with continuous light at different intensities. Exponential growth unlimited by gas transfer or self-shading was observable for a period of several days using fluorescence, which is an order of magnitude more sensitive than optical density. The microplate datasets were comparable to similar datasets obtained in photobioreactors and were used an input for the Huesemann model to accurately predict P(v). PMID:25103033

  20. 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

  1. 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.

  2. In Vitro Streptococcus pneumoniae Biofilm Formation and In Vivo Middle Ear Mucosal Biofilm in a Rat Model of Acute Otitis Induced by S. pneumoniae

    PubMed Central

    Yadav, Mukesh Kumar; Chae, Sung-Won

    2012-01-01

    Objectives Streptococcus pneumoniae is one of the most common pathogens of otitis media (OM) that exists in biofilm, which enhances the resistance of bacteria against antibiotic killing and diagnosis, compared to the free-floating (planktonic) form. This study evaluated biofilm formation by S. pneumoniae on an abiotic surface and in the middle ear cavity in a rat model of OM. Methods In vitro biofilm formation was evaluated by inoculation of a 1:100 diluted S. pneumoniae cell suspension in a 96-well microplate. Adherent cells were quantified spectrophotometrically following staining with crystal violet by measurement of optical density at 570 nm. The ultrastructure of pneumococcal biofilm was assessed by scanning electron microscopy (SEM). For in vitro biofilm study, S. pneumoniae cell suspensions containing 1×107 colony forming units were injected through transtympanic membrane into the middle ear cavity of Sprague Dawley rats. The ultrastructure of middle ear mucus was observed by SEM 1 and 2 weeks post-inoculation. Results The in vitro study revealed robust biofilm formation by S. pneumoniae after 12-18 hours of incubation in high glucose medium, independent of exogenously supplied competence stimulating peptide and medium replacement. Adherent cells formed three-dimensional structures approximately 20-30 µm thick. The in vivo study revealed that ciliated epithelium was relatively resistant to biofilm formation and that biofilm formation occurred mainly on non-ciliated epithelium of the middle ear cavity. One week after inoculation, biofilm formation was high in 50% of the treated rats and low in 25% of the rats. After 2 weeks, biofilm formation was high and low in 25% and 37.5% of rats, respectively. Conclusion The results imply that glucose level is important for the S. pneumoniae biofilm formation and S. pneumoniae biofilm formation may play important role in the pathophysiology of OM. PMID:22977710

  3. 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

  4. 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.

  5. 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

  6. 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

  7. Direct formation of thienopyridine-derived nitrosothiols--just add nitrite!

    PubMed

    Bundhoo, Shantu S; Anderson, Richard A; Sagan, Ewelina; Hassan, Nurudeen; Pinder, Andrew G; Rogers, Stephen C; Morris, Keith; James, Philip E

    2011-11-30

    Thienopyridines (ticlopidine, clopidogrel and prasugrel) are pro-drugs that require metabolism to exhibit a critical thiol group in the active form that binds to the P2Y₁₂ receptor to inhibit platelet activation and prevent thrombus formation in vivo. We investigated whether these thienopyridines participate in S-nitrosation (SNO) reactions that might exhibit direct anti-platelet behaviour. Optimum conditions for in vitro formation of thienopyridine-SNO formation were studied by crushing ticlopidine, clopidogrel or prasugrel into aqueous solution and adding sodium nitrite, or albumin-SNO. Ozone-based chemiluminescence techniques were utilised to specifically detect NO release from the SNO produced. Effect on agonist-induced platelet aggregation was monitored using light transmittance in a 96 well microplate assay. Pharmaceutical grade preparations of ticlopidine, clopidogrel and prasugrel were found to exhibit significant free thiol and formed SNO derivatives directly from anionic nitrite in water under laboratory conditions without the need for prior metabolism. Thienopyridine-SNO formation was dependent on pH, duration of mixing and nitrite concentration, with prasugrel-SNO being more favourably formed. The SNO moiety readily participated in trans-nitrosation reactions with albumin and plasma. Prasugrel-SNO showed significantly better inhibition of platelet aggregation compared with clopidogrel-SNO, however when compared on the basis of SNO concentration these were equally effective (IC₅₀=7.91 ± 1.03 v/s 10.56 ± 1.43 μM, ns). Thienopyridine-derived SNO is formed directly from the respective base drug without the need for prior in vivo metabolism and therefore may be an important additional contributor to the pharmacological effectiveness of thienopyridines not previously considered. PMID:21946108

  8. 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

  9. 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.

  10. "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

  11. 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.

  12. 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

  13. 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

  14. 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

  15. 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

  16. 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.

  17. 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

  18. 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

  19. 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

  20. Development of a pneumatically driven active cover lid for multi-well microplates for use in perfusion three-dimensional cell culture

    PubMed Central

    Huang, Song-Bin; Chou, Dean; Chang, Yu-Han; Li, Ke-Cing; Chiu, Tzu-Keng; Ventikos, Yiannis; Wu, Min-Hsien

    2015-01-01

    Before microfluidic-based cell culture models can be practically utilized for bioassays, there is a need for a transitional cell culture technique that can improve conventional cell culture models. To address this, a hybrid cell culture system integrating an active cover lid and a multi-well microplate was proposed to achieve perfusion 3-D cell culture. In this system, a microfluidic-based pneumatically-driven liquid transport mechanism was integrated into the active cover lid to realize 6-unit culture medium perfusion. Experimental results revealed that the flow of culture medium could be pneumatically driven in a flow-rate uniform manner. We used the system to successfully perform a perfusion 3-D cell culture of mesenchymal stem cells (MSCs) for up to 16 days. Moreover, we investigated the effects of various cell culture models on the physiology of MSCs. The physiological nature of MSCs can vary with respect to the cell culture model used. Using the perfusion 3-D cell culture format might affect the proliferation and osteogenic differentiation of MSCs. Overall, we have developed a cell culture system that can achieve multi-well microplate-based perfusion 3-D cell culture in an efficient, cost-effective, and user-friendly manner. These features could facilitate the widespread application of perfusion cell culture models for cell-based assays. PMID:26669749

  1. Development of a pneumatically driven active cover lid for multi-well microplates for use in perfusion three-dimensional cell culture

    NASA Astrophysics Data System (ADS)

    Huang, Song-Bin; Chou, Dean; Chang, Yu-Han; Li, Ke-Cing; Chiu, Tzu-Keng; Ventikos, Yiannis; Wu, Min-Hsien

    2015-12-01

    Before microfluidic-based cell culture models can be practically utilized for bioassays, there is a need for a transitional cell culture technique that can improve conventional cell culture models. To address this, a hybrid cell culture system integrating an active cover lid and a multi-well microplate was proposed to achieve perfusion 3-D cell culture. In this system, a microfluidic-based pneumatically-driven liquid transport mechanism was integrated into the active cover lid to realize 6-unit culture medium perfusion. Experimental results revealed that the flow of culture medium could be pneumatically driven in a flow-rate uniform manner. We used the system to successfully perform a perfusion 3-D cell culture of mesenchymal stem cells (MSCs) for up to 16 days. Moreover, we investigated the effects of various cell culture models on the physiology of MSCs. The physiological nature of MSCs can vary with respect to the cell culture model used. Using the perfusion 3-D cell culture format might affect the proliferation and osteogenic differentiation of MSCs. Overall, we have developed a cell culture system that can achieve multi-well microplate-based perfusion 3-D cell culture in an efficient, cost-effective, and user-friendly manner. These features could facilitate the widespread application of perfusion cell culture models for cell-based assays.

  2. Development of a pneumatically driven active cover lid for multi-well microplates for use in perfusion three-dimensional cell culture.

    PubMed

    Huang, Song-Bin; Chou, Dean; Chang, Yu-Han; Li, Ke-Cing; Chiu, Tzu-Keng; Ventikos, Yiannis; Wu, Min-Hsien

    2015-01-01

    Before microfluidic-based cell culture models can be practically utilized for bioassays, there is a need for a transitional cell culture technique that can improve conventional cell culture models. To address this, a hybrid cell culture system integrating an active cover lid and a multi-well microplate was proposed to achieve perfusion 3-D cell culture. In this system, a microfluidic-based pneumatically-driven liquid transport mechanism was integrated into the active cover lid to realize 6-unit culture medium perfusion. Experimental results revealed that the flow of culture medium could be pneumatically driven in a flow-rate uniform manner. We used the system to successfully perform a perfusion 3-D cell culture of mesenchymal stem cells (MSCs) for up to 16 days. Moreover, we investigated the effects of various cell culture models on the physiology of MSCs. The physiological nature of MSCs can vary with respect to the cell culture model used. Using the perfusion 3-D cell culture format might affect the proliferation and osteogenic differentiation of MSCs. Overall, we have developed a cell culture system that can achieve multi-well microplate-based perfusion 3-D cell culture in an efficient, cost-effective, and user-friendly manner. These features could facilitate the widespread application of perfusion cell culture models for cell-based assays. PMID:26669749

  3. 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

  4. 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

  5. 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.

  6. 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

  7. 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

  8. 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

  9. 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.

  10. 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

  11. 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.

  12. 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.

  13. 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...

  14. 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

  15. 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

  16. 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

  17. 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.

  18. 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

  19. 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

  20. 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

  1. 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

  2. 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

  3. 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

  4. 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.

  5. 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.

  6. 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

  7. 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

  8. 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

  9. Paleomagnetism, Geochronology, and Geochemistry of the Type Section of the Stanislaus Group: Reference Parameters from the Stable Sierra Nevada Microplate, CA

    NASA Astrophysics Data System (ADS)

    Farner, M. J.; Pluhar, C. J.; Asami, R.; Putirka, K. D.; Busby, C.; Renne, P. R.

    2012-12-01

    The Late Miocene Stanislaus Group, of California and Nevada is composed of Table Mountain Formation, Eureka Valley Tuff, and Dardanelles Formation. This ~9.0-~10.2 million year old unit interrupted Miocene andesitic arc volcanism in the Sierra Nevada, providing a regional lithostratigraphic marker that has been used extensively to reconstruct tilt and uplift of the range, Neogene tectonics of the Walker Lane Belt, magmagenetic processes beneath the Sierra Nevada, and lithospheric evolution of the Sierra Nevada and Eastern California. A recent study (Koerner et al, 2009) produced a measured section and geologic map of the Stanislaus Group type section, but until now this locality has never seen comprehensive multidisciplinary study of the geochronology, geochemistry, and magnetostratigraphy of the site and to integrate this into the overall understanding of the Stanislaus Group. Stratigraphy, geochemistry, and paleomagnetism from the type section suggest addition of a basal trachyte lava flow member to the Eureka Valley tuff and adds an additional intermediate-polarity lava flow to Table Mountain Formation magnetostratigraphy. This study dates the youngest member of the Stanislaus Group, the Dardanelles Formation, by 40Ar/39Ar radioisotopic dating for the first time, yielding an age of 9.048 ± 0.017 Ma. Paleomagnetic results verify the previous paleomagnetic reference direction from the Sierra Nevada microplate for the Tollhouse Flat Member of the Eureka Valley Tuff (King et al., 2007). However, our work revises the By-Day Member reference direction to D = 349.6°, I = 51.9° n = 8, α95 = 3.0°, k = 346. This difference is because the prior work analyzed By-Day localities within the tectonically-active Walker Lane Belt. The revised reference direction is critical for measurements of relative vertical-axis rotation studies in the Walker Lane. Our study also demonstrates that little to no vertical-axis rotation of the Sierra Nevada microplate has occurred since

  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. 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.

  15. 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

  16. 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

  17. 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.

  18. 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

  19. 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

  20. 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.

  1. 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.

  2. 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

  3. An outer ramp to basin plain transect: Interacting pelagic and calciturbidite deposition in the Eocene-Oligocene of the Tuscan Domain, Adria Microplate (Italy)

    NASA Astrophysics Data System (ADS)

    Ielpi, Alessandro; Cornamusini, Gianluca

    2013-08-01

    The interaction of ramps, basin plains and turbidite systems on the scale of tens of km has been rarely observed in fossil examples. Deep marine Eocene-Oligocene beds are exposed in the axial zone of the Chianti Mountains, Italy, and compose a regionally continue stratigraphic succession known as the Scaglia Toscana Formation. The formation was deposited in the Tuscan Domain of the Adria Microplate. This research aims at depicting its depositional architecture and evolution in the type area. Stratigraphic and sedimentologic analyses were performed on a ca. 25 km-long transect that includes depositional systems sectioned both in the down- and along-dip directions. Shaly-carbonate deposits compose a complex of interacting ramps, basin plains and turbidite floor fan systems. Ramp deposits accumulated above the lysocline and in oxic conditions. Basin plain beds were deposited below the lysocline and were subject to episodes of oxygen depletion. Turbidity flows fed elongate fan lobes characterized by poor channelisation. The basin palaeogeography hampered the development of slope apron turbidite systems. The Eocene-Oligocene geodynamic setting of the Tuscan Domain was characterized by the evolution of a peripheral bulge and by the early structuring of a foredeep basin. Syn-sedimentary tectonism acted a primary role in the basin-scale arrangement. However other mechanisms also contributed to the local facies distribution, including the disposition of sediment-source areas and intrabasinal confinement morphologies, as well as relative oscillations of the depositional surface with respect to the lysocline and oxycline.

  4. 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.

  5. 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

  6. 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

  7. 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

  8. 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.

  9. 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

  10. 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

  11. 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.

  12. 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

  13. 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

  14. 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

  15. 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

  16. 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

  17. 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

  18. 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

  19. 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

  20. 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

  1. 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

  2. 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

  3. Novel Time-Resolved Fluorescence Europium Nanoparticle Immunoassay for Detection of Human Immunodeficiency Virus-1 Group O Viruses Using Microplate and Microchip Platforms.

    PubMed

    Haleyur Giri Setty, Mohan Kumar; Liu, Jikun; Mahtani, Prerna; Zhang, Panhe; Du, Bingchen; Ragupathy, Viswanath; Devadas, Krishnakumar; Hewlett, Indira K

    2016-06-01

    Accurate detection and quantification of HIV-1 group O viruses have been challenging for currently available HIV assays. We have developed a novel time-resolved fluorescence (TRF) europium nanoparticle immunoassay for HIV-1 group O detection using a conventional microplate enzyme-linked immunosorbent assay (ELISA) and a microchip platform. We screened several antibodies for optimal reactivity with several HIV-1 group O strains and identified antibodies that can detect all the strains of HIV-1 group O that were available for testing. The antibodies were used to develop a conventional ELISA format assay and an in-house developed europium nanoparticle-based assay for sensitivity. The method was evaluated on both microwell plate and microchip platforms. We identified two specific and sensitive antibodies among the six we screened. The antibodies, C65691 and ANT-152, were able to quantify 15 and detect all 17 group O viruses, respectively, as they were broadly cross-reactive with all HIV-1 group O strains and yielded better signals compared with other antibodies. We have developed a sensitive assay that reflects the actual viral load in group O samples by using an appropriate combination of p24 antibodies that enhance group O detection and a highly sensitive TRF-based europium nanoparticle for detection. The combination of ANT-152 and C65690M in the ratio 3:1 was able to give significantly higher signals in our europium-based assay compared with using any single antibody. PMID:26978478

  4. 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

  5. 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

  6. 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

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

    PubMed

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

    2010-04-01

    Microplitis mediator Haliday (Hymenoptera: Braconidae) is an important larval endoparasitoid of various lepidopteran pests, including Helicoverpa armigera (Hübner). In China, H. armigera is a key pest of cotton and is currently the focus of several biological control efforts that use M. mediator as principal natural enemy of this pest. To improve the success of biological control efforts, behavioral studies are needed that shed light on the interaction between M. mediator and H. armigera. In this study, we determined M. mediator response to volatile compounds from undamaged, mechanically injured, or H. armigera--damaged plants and identified attractive volatiles. In Y-tube olfactometer assays, we found that mechanically damaged plants and/or plants treated with H. armigera oral secretions did not attract wasps. However, volatiles from H. armigera-damaged plants elicited a strong attraction of both M. mediator sexes. Headspace extracts from H. armigera-damaged cotton were analyzed by coupled gas chromatography-electroantennographic detection (GC-EAD), and a total of seven different compounds were found to elicit electroantennogram (EAG) responses, including an unknown compound. Six different EAD-active volatiles were identified from caterpillar-damaged cotton plants, of which 3, 7-dimethyl-1, 3, 6-octatriene and (Z)-3-hexenyl acetate were the principal compounds. Olfactometer assays indicated that individual synthetic compounds of 3, 7-dimethyl-1, 3, 6-octatriene, (Z)-3-hexenyl acetate, and nonanal were attractive to M. mediator. Field cage studies showed that parasitism of H. armigera larvae by M. mediator was higher on cotton plants to which 3,7-dimethyl-1,3, 6-octatriene was applied. Our results show that the combination of terpenoids and green leaf volatiles may not only facilitate host, mate, or food location but may also increase H. armigera parasitism by M. mediator. PMID:20388293

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

    SciTech Connect

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

    2007-03-01

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

  9. 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

  10. 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

  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. 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.

  14. 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

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

    PubMed

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

    2009-03-01

    Developing enzyme cocktails for cellulosic biomass hydrolysis complementary to current cellulase systems is a critical step needed for economically viable biofuels production. Recent genomic analysis indicates that some plant pathogenic fungi are likely a largely untapped resource in which to prospect for novel hydrolytic enzymes for biomass conversion. In order to develop high throughput screening assays for enzyme bioprospecting, a standardized microplate assay was developed for rapid analysis of polysaccharide hydrolysis by fungal extracts, incorporating biomass substrates. Fungi were grown for 10 days on cellulose- or switchgrass-containing media to produce enzyme extracts for analysis. Reducing sugar released from filter paper, Avicel, corn stalk, switchgrass, carboxymethylcellulose, and arabinoxylan was quantified using a miniaturized colorimetric assay based on 3,5-dinitrosalicylic acid. Significant interactions were identified among fungal species, growth media composition, assay substrate, and temperature. Within a small sampling of plant pathogenic fungi, some extracts had crude activities comparable to or greater than T. reesei, particularly when assayed at lower temperatures and on biomass substrates. This microplate assay system should prove useful for high-throughput bioprospecting for new sources of novel enzymes for biofuel production. PMID:18973283

  16. 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.

  17. 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

  18. 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

  19. 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

  20. 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

  1. 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

  2. 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

  3. 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

  4. 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

  5. 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.

  6. 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

  7. 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.

  8. High-throughput functional screening of steroid substrates with wild-type and chimeric P450 enzymes.

    PubMed

    Urban, Philippe; Truan, Gilles; Pompon, Denis

    2014-01-01

    The promiscuity of a collection of enzymes consisting of 31 wild-type and synthetic variants of CYP1A enzymes was evaluated using a series of 14 steroids and 2 steroid-like chemicals, namely, nootkatone, a terpenoid, and mifepristone, a drug. For each enzyme-substrate couple, the initial steady-state velocity of metabolite formation was determined at a substrate saturating concentration. For that, a high-throughput approach was designed involving automatized incubations in 96-well microplate with sixteen 6-point kinetics per microplate and data acquisition using LC/MS system accepting 96-well microplate for injections. The resulting dataset was used for multivariate statistics aimed at sorting out the correlations existing between tested enzyme variants and ability to metabolize steroid substrates. Functional classifications of both CYP1A enzyme variants and steroid substrate structures were obtained allowing the delineation of global structural features for both substrate recognition and regioselectivity of oxidation. PMID:25243177

  9. High-Throughput Functional Screening of Steroid Substrates with Wild-Type and Chimeric P450 Enzymes

    PubMed Central

    Truan, Gilles; Pompon, Denis

    2014-01-01

    The promiscuity of a collection of enzymes consisting of 31 wild-type and synthetic variants of CYP1A enzymes was evaluated using a series of 14 steroids and 2 steroid-like chemicals, namely, nootkatone, a terpenoid, and mifepristone, a drug. For each enzyme-substrate couple, the initial steady-state velocity of metabolite formation was determined at a substrate saturating concentration. For that, a high-throughput approach was designed involving automatized incubations in 96-well microplate with sixteen 6-point kinetics per microplate and data acquisition using LC/MS system accepting 96-well microplate for injections. The resulting dataset was used for multivariate statistics aimed at sorting out the correlations existing between tested enzyme variants and ability to metabolize steroid substrates. Functional classifications of both CYP1A enzyme variants and steroid substrate structures were obtained allowing the delineation of global structural features for both substrate recognition and regioselectivity of oxidation. PMID:25243177

  10. 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

  11. 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

  12. 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

  13. 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

  14. 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

  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. 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.

  18. 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

  19. 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

  20. 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.

  1. 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.

  2. 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

  3. 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

  4. 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.

  5. 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

  6. Interaction of the Koinobiont Parasitoid Microplitis rufiventris of the Cotton Leafworm, Spodoptera littoralis, with Two Entomopathogenic Rhabditids, Heterorhabditis bacteriophora and Steinernema carpocapsae

    PubMed Central

    Atwa, Atwa A.; Hegazi, Esmat M.; Khafagi, Wedad E.; El-Aziz, Gehan M. Abd

    2013-01-01

    Entomopathogenic nematodes are generally considered beneficial organisms. However, they can affect beneficial insects such as parasitoids. The interaction between the entomopathogenic nematodes Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae) and Steinernema carpocapsae Weiser, and the parasitoid Microplitis rufiventris Kokujev (Hymenoptera: Braconidae) was investigated in the laboratory. In non-parasitized hosts, Spodoptera littoralis Boisduval (Lepidoptera: Noctuidae) larvae exposed to H. bacteriophora showed a higher percent mortality than those exposed to S. carpocapsae. Both nematodes were able to invade and propagate in non-parasitized S. littoralis larvae and those parasitized by M. rufiventris. Both nematode species reproduced in Microplitis-parasitized hosts, but there was a higher number of nematodes in non-parasitized larvae. S. carpocapsae yielded higher numbers of infective juveniles than H. bacteriophora. Generally, the number of nematodes harvested increased as their host's size increased. The interaction between the nematodes and parasitoid favored the nematodes when the nematodes were inoculated during the parasitoid egg stage or the young parasitoid larvae, thus giving the nematodes a better chance to grow and reproduce, resulting in the death of the parasitoid larvae. Conversely, when the nematodes were inoculated during the late larval instar of the parasitoid, the competition partially favored the wasp, thus giving approximately 50% of the wasps a better chance to develop, emerge, and reproduce, providing evidence that both nematodes and wasps could reproduce in the same host. Egg maturation of female wasps derived from nematode-infected hosts was not significantly different than those from control hosts. The combined application of nematodes and parasitoids may be beneficial if the detrimental effects of the nematodes on the parasitoid could be avoided by precisely timing the application strategies. It is clear that

  7. Comparative cytotoxic and genotoxic potential of 13 drinking water disinfection by-products using a microplate-based cytotoxicity assay and a developed SOS/umu assay.

    PubMed

    Zhang, Shao-Hui; Miao, Dong-Yue; Tan, Li; Liu, Ai-Lin; Lu, Wen-Qing

    2016-01-01

    The implications of disinfection by-products (DBPs) present in drinking water are of public health concern because of their potential mutagenic, carcinogenic and other toxic effects on humans. In this study, we selected 13 main DBPs found in drinking water to quantitatively analyse their cytotoxicity and genotoxicity using a microplate-based cytotoxicity assay and a developed SOS/umu assay in Salmonella typhimurium TA1535/pSK1002. With the developed SOS/umu test, eight DBPs: 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-fura3-chloro-4-(dichloromethyl)-5-hydroxy-2-[5H]-furanone (MX), dibromoacetonitrile (DBN), iodoacetic acid (IA), bromochloroacetonitrile (BCN), bromoacetic acid (BA), trichloroacetonitrile (TCN), dibromoacetic acid (DBA) and dichloroacetic acid (DCA) were significantly genotoxic to S. typhimurium. Three DBPs: chloroacetic acid (CA), trichloroacetic acid (TCA) and dichloroacetonitrile (DCN) were weakly genotoxic, whereas the remaining DBPs: chloroacetonitrile (CN) and chloral hydrate (CH) were negative. The rank order in decreasing genotoxicity was as follows: MX > DBN > IA > BCN > BA > TCN > DBA > DCA > CA, TCA, DCN > CN, CH. MX was approximately 370 000 times more genotoxic than DCA. In the microplate-based cytotoxicity assay, cytotoxic potencies of the 13 DBPs were compared and ranked in decreasing order as follows: MX > IA > DBN > BCN > BA > TCN > DCN > CA > DCA > DBA > CN > TCA > CH. MX was approximately 19 200 times more cytotoxic than CH. A statistically significant correlation was found between cytotoxicity and genotoxicity of the 13 DBPs in S. typhimurium. Results suggest that microplate-based cytotoxicity assay and the developed SOS/umu assay are feasible tools for analysing the cytotoxicity and genotoxicity of DBPs, particularly for comparing their toxic intensities quantitatively. PMID:26188195

  8. Rapid Detection of an ABT-737-Sensitive Primed for Death State in Cells Using Microplate-Based Respirometry

    PubMed Central

    Clerc, Pascaline; Carey, Gregory B.; Mehrabian, Zara; Wei, Michael; Hwang, Hyehyun; Girnun, Geoffrey D.; Chen, Hegang; Martin, Stuart S.; Polster, Brian M.

    2012-01-01

    Cells that exhibit an absolute dependence on the anti-apoptotic BCL-2 protein for survival are termed “primed for death” and are killed by the BCL-2 antagonist ABT-737. Many cancers exhibit a primed phenotype, including some that are resistant to conventional chemotherapy due to high BCL-2 expression. We show here that 1) stable BCL-2 overexpression alone can induce a primed for death state and 2) that an ABT-737-induced loss of functional cytochrome c from the electron transport chain causes a reduction in maximal respiration that is readily detectable by microplate-based respirometry. Stable BCL-2 overexpression sensitized non-tumorigenic MCF10A mammary epithelial cells to ABT-737-induced caspase-dependent apoptosis. Mitochondria within permeabilized BCL-2 overexpressing cells were selectively vulnerable to ABT-737-induced cytochrome c release compared to those from control-transfected cells, consistent with a primed state. ABT-737 treatment caused a dose-dependent impairment of maximal O2 consumption in MCF10A BCL-2 overexpressing cells but not in control-transfected cells or in immortalized mouse embryonic fibroblasts lacking both BAX and BAK. This impairment was rescued by delivering exogenous cytochrome c to mitochondria via saponin-mediated plasma membrane permeabilization. An ABT-737-induced reduction in maximal O2 consumption was also detectable in SP53, JeKo-1, and WEHI-231 B-cell lymphoma cell lines, with sensitivity correlating with BCL-2:MCL-1 ratio and with susceptibility (SP53 and JeKo-1) or resistance (WEHI-231) to ABT-737-induced apoptosis. Multiplexing respirometry assays to ELISA-based determination of cytochrome c redistribution confirmed that respiratory inhibition was associated with cytochrome c release. In summary, cell-based respiration assays were able to rapidly identify a primed for death state in cells with either artificially overexpressed or high endogenous BCL-2. Rapid detection of a primed for death state in individual cancers

  9. Susceptibility of Mycobacterium tuberculosis to first-line antimycobacterial agents in a Brazilian hospital: assessing the utility of the tetrazolium (MTT) microplate assay.

    PubMed

    Ferrari, Michela De Luca; Telles, Maria Alice da Silva; Ferrazoli, Lucilaine; Levy, Carlos Emílio; Villares, Maria Cecília Barison; Moretti, Maria Luiza; Resende, Mariângela Ribeiro

    2010-08-01

    We conducted a cross-sectional, hospital-based study between January 2006-March 2008 to estimate the resistance of Mycobacterium tuberculosis to first-line drugs in patients with tuberculosis at a Brazilian hospital. We evaluated the performance of the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] (MTT) microplate assay compared with the Bactec-MGIT 960 system for mycobacteria testing. The prevalence of resistance in M. tuberculosis was 6.7%. Multidrug-resistance [resistance to rifampicin (RMP) and isoniazid (INH)], INH-resistance and streptomycin (SM)-resistance accounted for 1%, 3.8% and 3.8% of all resistance, respectively, and all isolates were susceptible to ethambutol (EM). The resistance was primary in four cases and acquired in three cases and previous treatment was associated with resistance (p = 0.0129). Among the 119 M. tuberculosis isolates, complete concordance of the results for INH and EM was observed between the MTT microplate and Bactec-MGIT 960TM methods. The observed agreement for RMP was 99% (sensitivity: 90%) and 95.8% for SM (sensitivity 90.9%), lower than those for other drugs. The MTT colourimetric method is an accurate, simple and low-cost alternative in settings with limited resources. PMID:20835613

  10. A New Barrier to Dispersal Trapped Old Genetic Clines That Escaped the Easter Microplate Tension Zone of the Pacific Vent Mussels

    PubMed Central

    Plouviez, Sophie; Faure, Baptiste; Le Guen, Dominique; Lallier, François H.; Bierne, Nicolas; Jollivet, Didier

    2013-01-01

    Comparative phylogeography of deep-sea hydrothermal vent species has uncovered several genetic breaks between populations inhabiting northern and southern latitudes of the East Pacific Rise. However, the geographic width and position of genetic clines are variable among species. In this report, we further characterize the position and strength of barriers to gene flow between populations of the deep-sea vent mussel Bathymodiolus thermophilus. Eight allozyme loci and DNA sequences of four nuclear genes were added to previously published sequences of the cytochrome c oxidase subunit I gene. Our data confirm the presence of two barriers to gene flow, one located at the Easter Microplate (between 21°33′S and 31°S) recently described as a hybrid zone, and the second positioned between 7°25′S and 14°S with each affecting different loci. Coalescence analysis indicates a single vicariant event at the origin of divergence between clades for all nuclear loci, although the clines are now spatially discordant. We thus hypothesize that the Easter Microplate barrier has recently been relaxed after a long period of isolation and that some genetic clines have escaped the barrier and moved northward where they have subsequently been trapped by a reinforcing barrier to gene flow between 7°25′S and 14°S. PMID:24312557

  11. Determining cereal starch amylose content using a dual wavelength iodine binding 96 well plate assay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cereal starch amylose/amylopectin (AM/AP) ratios are critical in functional properties for food and industrial applications. Conventional determination of AM/AP of cereal starches are very time consuming and labor intensive making it very difficult to screen large sample sets. Studying these large...

  12. Development of a 96-well plate iodine binding assay for amylose content determination

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cereal starch amylose/amylopectin (AM/AP) ratios are critical in functional properties for food and industrial applications. Conventional methods for the determination of AM/AP of cereal starches are very time consuming and labor intensive making it very difficult to screen large sample sets. Stud...

  13. Folate content in fresh-cut vegetable packed products by 96-well microtiter plate microbiological assay.

    PubMed

    Fajardo, Violeta; Alonso-Aperte, Elena; Varela-Moreiras, Gregorio

    2015-02-15

    Ready-to-eat foods have nowadays become a significant portion of the diet. Accordingly, nutritional composition of these food categories should be well-known, in particular its folate content. However, there is a broad lack of folate data in food composition tables and databases. A total of 21 fresh-cut vegetable and fruit packed products were analysed for total folate (TF) content using a validated method that relies on the folate-dependent growth of chloramphenicol-resistant Lactobacillus casei subspecies rhamnosus (NCIMB 10463). Mean TF content ranged from 10.0 to 140.9μg/100g for the different matrices on a fresh weight basis. Higher TF quantity, 140.9-70.1μg/100g, was found in spinach, rocket, watercress, chard and broccoli. Significant differences were observed between available data for fresh vegetables and fruits from food composition tables or databases and the analysed results for fresh-cut packed products. Supplied data support the potential of folate-rich fresh-cut ready-to-eat vegetables to increase folate intake significantly. PMID:25236228

  14. Is biofilm removal properly assessed? Comparison of different quantification methods in a 96-well plate system.

    PubMed

    Stiefel, Philipp; Rosenberg, Urs; Schneider, Jana; Mauerhofer, Stefan; Maniura-Weber, Katharina; Ren, Qun

    2016-05-01

    Various methods have been reported to quantify total biofilm or different components of biofilm; however, these methods are often confusedly used, leading to discrepancies and misleading results. In this study, different methods for quantification of biofilm, including those for total biomass, total amount of bacterial cells, viable cell number, and amount of extracellular polymeric substances, were systematically compared in microtiter plates. To evaluate which method is suitable for assessment of biofilm removal and for bacterial killing, biofilm samples were treated with various cleaners possessing removing and/or killing capacities. It was found that most of the methods tested in this study in general exhibited high reproducibility and repeatability. Crystal Violet staining was a simple but reliable method for total biomass quantification. Total bacteria cell numbers could be reliably quantified by the fluorescent DNA-binding dye Acridine Orange. Viable cells could be quantified by either an ATP-based assay or a proliferation assay. Both of these viability methods showed a broad detection range and led to precise measurement. For quantification of proteins in the biofilm, staining with fluorescein isothiocyanate was most suitable. Furthermore, it was revealed that a combination of different methods is required to determine if a cleaner kills or removes biofilm. PMID:26923144

  15. Individualized Surgical Templates and Titanium Microplates for Le Fort I Osteotomy by Computer-Aided Design and Computer-Aided Manufacturing.

    PubMed

    He, Wei; Tian, Kaiyue; Xie, Xiaoyan; Wang, Xiaoxia; Li, Yang; Wang, Xing; Li, Zili

    2015-09-01

    The authors report the use of novel individualized surgical templates and titanium miniplates for Le Fort I osteotomy and evaluate the accuracy of this technique in vitro. Nine three-dimensional stereolithographic skull models were used to design the templates and titanium microplates and to simulate the operation. Cone beam computed tomography (CBCT) scans of the skulls were acquired preoperatively and were used to generate virtual models. The surgical plans were made based on three-dimensional cephalometric analyses, and osteotomies were then performed virtually. Cylinder-shaped markers were placed to permit the correct location of titanium screws, and individualized surgical templates were designed. The bony segments were then repositioned virtually according to the surgical plans to correct the skeletal deformities. Resin surgical templates were produced by stereolithography rapid prototyping and the titanium miniplates by three-dimensional cutting. Le Fort I osteotomy was performed under the guide of the surgical templates and fixed with the titanium miniplates. Postoperatively, CBCT scans of each skull model were taken, and the differences between the actual and planned surgical outcomes were measured by superimposing the planned and postoperative virtual models generated from CBCT images. The authors demonstrated that the average linear difference between the planned and actual outcomes was <1 mm and the average orientation difference was <1°. The individualized surgical templates and titanium microplates designed in this experimental study permitted the repositioning of the maxillary segment to the correct planned positions during Le Fort I osteotomy, making this technique a promising alternative to the conventional split method. PMID:26221851

  16. Formation of different gold nanostructures by silk nanofibrils.

    PubMed

    Fang, Guangqiang; Yang, Yuhong; Yao, Jinrong; Shao, Zhengzhong; Chen, Xin

    2016-07-01

    Metal nanostructures that have unique size- and shape-dependent electronic, optical and chemical properties gain more and more attention in modern science and technology. In this article, we show the possibility that we are able to obtain different gold nanostructures simply with the help of silk nanofibrils. We demonstrate that only by varying the pH of the reaction solution, we get gold nanoparticles, nano-icosahedrons, nanocubes, and even microplates. Particularly, we develop a practical method for the preparation of gold microplates in acid condition in the presence of silk nanofibrils, which is impossible by using other forms of silk protein. We attribute the role of silk nanofibrils in the formation of gold nanostructure to their reduction ability from several specific amino acid residues, and the suitable structural anisotropic features to sustain the crystal growth after the reduction process. Although the main purpose of this article is to demonstrate that silk nanofibrils are able to mediate the formation of different gold nanostructure, we show the potential applications of these resulting gold nanostructures, such as surface-enhanced Raman scattering (SERS) and photothermal transformation effect, as same as those produced by other methods. In conclusion, we present in this communication a facile and green synthesis route to prepare various gold nanostructures with silk nanofibrils by simply varying pH in the reaction system, which has remarkable advantages in future biomedical applications. PMID:27127067

  17. Highly sensitive micro-plate enzyme immunoassay screening and NCI-GC-MS confirmation of flunitrazepam and its major metabolite 7-aminoflunitrazepam in hair.

    PubMed

    Negrusz, A; Moore, C; Deitermann, D; Lewis, D; Kaleciak, K; Kronstrand, R; Feeley, B; Niedbala, R S

    1999-10-01

    Flunitrazepam (Rohypnol) is a benzodiazepine used in the treatment of insomnia as a sedative hypnotic and as preanesthetic medication in European countries and Mexico. Although it has no medicinal purpose in the United States, the occurrence of its abuse is increasing. Sexual abuse of both men and women while under the influence of so-called "date-rape" drugs has been the focus of many investigations. Reported date-rape drugs include flunitrazepam (FN), clonazepam, diazepam, oxazepam, gamma-hydroxybutyrate, and many others. FN has been banned in the United States because of its alleged use in such situations. Unfortunately, the detection of FN or its metabolites 7-aminoflunitrazepam (7-AFN) and desmethylflunitrazepam in a single specimen such as urine or blood is difficult in criminal situations because of the likelihood of single-dose ingestion and the length of time since the alleged incident. Hair provides a solution to the second of these problems in that drugs tend to incorporate into hair and remain there for longer periods of time than either urine or blood. There are various techniques for the detection of FN in plasma, blood, and urine, but little work has been done with hair. Hair collection is a virtually noninvasive procedure that can supply information on drug use for several months preceding collection. The objective of this paper was to determine if a commercially available micro-plate enzyme immunoassay system was sufficiently sensitive for the routine screening of 7-AFN in hair by the development of extraction procedures and optimization of the immunoassay kit. Further, this study used the same solid-phase extraction to isolate FN and its major metabolite, 7-AFN, and gas chromatography-mass spectrometry with negative ion chemical ionization for confirmation. Two seven-point standard curves were established ranging from 0.5 pg/mg to 100 pg/mg for 7-AFN and 2.5 pg/mg to 200 pg/mg for FN with respective deuterated internal standards. A replicate

  18. Thermal conditions of the central Sinai Microplate inferred from new surface heat-flow values and continuous borehole temperature logging in central and southern Israel

    NASA Astrophysics Data System (ADS)

    Schütz, F.; Förster, H.-J.; Förster, A.

    2014-05-01

    This paper reports ten new surface heat-flow density (qs) values for central and southern Israel (central Sinai Microplate), whose crystalline crust and lithosphere formed as part of the Neoproterozoic Arabian-Nubian Shield. Heat flow was calculated in Mesozoic sediments using the classical approach of heat-flow determination by implementing in the analysis high-precision continuous temperature logs obtained in air- and/or water-filled boreholes. Thermal conductivity (TC) measured for a large suite of rock samples of lithotypes making up the sequence was assigned to temperature gradients in intervals for which the lithology was known. The heat-flow values obtained for different depth intervals in a borehole as well as the average values for the individual borehole locations cover a narrow range, attesting heat-conduction conditions. A steady-state thermal model along an E-W crustal cross section through the area shows that the observed systematic spatial distribution of the qs values, which range between 50 and 62 mW m-2, can primarily be explained by variations in the thickness of the upper crust and in the ratio between sedimentary and crystalline rocks therein. Given the time lapse of thermal heat transfer through the lithosphere, the qs data monitor the crustal thermal conditions prior to rift- and plume-related lithospheric thermal perturbations that have started in the larger area ca. 30 Ma ago. Observed and modeled qs display the best fit for a pre-Oligocene lithosphere-asthenosphere boundary (LAB) at ˜150 km, which would be at the upper end of LAB depths determined from stable areas of the Arabian Shield (150-120 km) not affected by the young, deep-seated thermal processes that have caused a further uprise of the LAB. Our data imply or predict that the surface heat flow of the Sinai Microplate generally tends to increase along N-S and W-E traverses, from ˜45-50 mW m-2 to ˜55-60 mW m-2. Surface heat flows on the order of 55-60 mW m-2 may be common in the

  19. Tectonic geomorphology and paleoseismology of strike-slip faults in Jamaica: Implications for distribution of strain and seismic hazard along the southern edge of the Gonave microplate

    NASA Astrophysics Data System (ADS)

    Koehler, R. D.; Mann, P.; Brown, L. A.

    2009-12-01

    The east-west, left lateral strike-slip fault system forming the southern edge of the Gonave microplate crosses the110-km-long and 70-km-wide island of Jamaica. GPS measurements in the northeastern Caribbean are supportive of the microplate interpretation and indicate that ~ half of the Caribbean-North America left-lateral plate motion (8-14 mm/yr) is carried by the Plantain Garden (PGFZ) and associated faults in Jamaica. We performed Neotectonic mapping of the Plantain Garden fault along the southern rangefront of the Blue Mountains and conducted a paleoseismic study of the fault at Morant River. Between Holland Bay and Morant River, the fault is characterized by a steep, faceted, linear mountain front, prominent linear valleys and depressions, shutter ridges, and springs. At the eastern end of the island, the PGFZ is characterized by a left-stepping fault geometry that includes a major, active hot spring. The river cut exposure at Morant River exposes a 1.5-m-wide, sub-vertical fault zone juxtaposing sheared alluvium and faulted Cretaceous basement rocks. This section is overlain by an, unfaulted 3-m-thick fluvial terrace inset into a late Pleistocene terrace that is culturally modified. Upward fault terminations indicate the occurrence of three paleoearthquakes that occurred prior to deposition of the flat lying inset terrace around 341-628 cal yr BP. At this time, our radiocarbon results suggest that we can rule out the PGFZ as the source of the 1907 Kingston earthquake 102 years ago, as well as, the 1692 event that destroyed Port Royal 317 years ago and produced a major landslide at Yallahs. Pending OSL ages will constrain the age of the penultimate and most recent ruptures. Gently to steeply dipping rocks as young as Pliocene exposed in roadcuts within the low coastal hills south of and parallel to the Plantain Garden fault may indicate active folding and blind thrust faulting. These structures are poorly characterized and may accommodate an unknown amount of

  20. Coupling HPLC-SPE-NMR with a microplate-based high-resolution antioxidant assay for efficient analysis of antioxidants in food--validation and proof-of-concept study with caper buds.

    PubMed

    Wiese, Stefanie; Wubshet, Sileshi G; Nielsen, John; Staerk, Dan

    2013-12-15

    This work describes the coupling of a microplate-based antioxidant assay with a hyphenated system consisting of high-performance liquid chromatography-solid-phase extraction-nuclear magnetic resonance spectroscopy, i.e., HPLC-SPE-NMR/high-resolution antioxidant assay, for the analysis of complex food extracts. The applicability of the microplate-based antioxidant assay for high-resolution screening of common food phenolics as well as parameters related to their trapping efficiency, elution behavior, and recovery on/from SPE cartridges are described. It was found that the microplate-based high-resolution antioxidant assay is an attractive and easy implementable alternative to direct on-line screening methods. Furthermore, it was shown that Resin SH and Resin GP SPE material are superior to RP C18HD for trapping of phenolic compounds. Proof-of-concept study was performed with caper bud extract, revealing the most important antioxidants to be quercetin, kaempferol, rutin, kaempferol-3-O-β-rutinoside and N(1),N(5),N(10)-triphenylpropenoyl spermidine amides. Targeted isolation of the latter, and comprehensive NMR experiments showed them to be N(1),N(10)-di-(E)-caffeoyl-N(5)-p-(E)-coumaroyl spermidine, N(1)-(E)-caffeoyl-N(5),N(10)-di-p-(E)-coumaroyl spermidine, N(10)-(E)-caffeoyl-N(1),N(5)-di-p-(E)-coumaroyl spermidine, and N(1),N(5),N(10)-tri-p-(E)-coumaroyl spermidine amides. PMID:23993578

  1. Analysis of dextromethorphan and dextrorphan in decomposed skeletal tissues by microwave assisted extraction, microplate solid-phase extraction and gas chromatography- mass spectrometry (MAE-MPSPE-GCMS).

    PubMed

    Fraser, Candice D; Cornthwaite, Heather M; Watterson, James H

    2015-08-01

    Analysis of decomposed skeletal tissues for dextromethorphan (DXM) and dextrorphan (DXT) using microwave assisted extraction (MAE), microplate solid-phase extraction (MPSPE) and gas chromatography-mass spectrometry (GC-MS) is described. Rats (n = 3) received 100 mg/kg DXM (i.p.) and were euthanized by CO2 asphyxiation roughly 20 min post-dose. Remains decomposed to skeleton outdoors and vertebral bones were recovered, cleaned, and pulverized. Pulverized bone underwent MAE using methanol as an extraction solvent in a closed microwave system, followed by MPSPE and GC-MS. Analyte stability under MAE conditions was assessed and found to be stable for at least 60 min irradiation time. The majority (>90%) of each analyte was recovered after 15 min. The MPSPE-GCMS method was fit to a quadratic response (R(2)  > 0.99), over the concentration range 10-10 000 ng⋅mL(-1) , with coefficients of variation <20% in triplicate analysis. The MPSPE-GCMS method displayed a limit of detection of 10 ng⋅mL(-1) for both analytes. Following MAE for 60 min (80 °C, 1200 W), MPSPE-GCMS analysis of vertebral bone of DXM-exposed rats detected both analytes in all samples (DXM: 0.9-1.5 µg⋅g(-1) ; DXT: 0.5-1.8 µg⋅g(-1) ). PMID:25487525

  2. Quaternary Expression of Northern Great Valley Faults and Folds: Accommodating North-South Contraction between the Sierran Microplate and Oregon Block

    NASA Astrophysics Data System (ADS)

    Angster, S.; Sawyer, T. L.; Wesnousky, S. G.

    2015-12-01

    The Northern California Shear Zone accommodates North American intraplate right-lateral transpressional shear driven by the relative motion of the northwest translating Sierran microplate. Within this zone, between the latitudes of 400 and 420, 1 - 4 mm/yr of north-south geodetic contraction is observed and for the most part remains geologically unaccounted for. We are investigating the Quaternary expression of transpressional shear localized at the northern end of the Sierra Nevada and Great Valley to evaluate both the recency and rates of deformation across structures that may be accommodating the contractional deformation suggested in geodetic studies. The northeast trending Inks Creek fold belt north of Red Bluff, CA consists of anticline-syncline pairs and dome structures that appear optimally oriented to accommodate northwest crustal shortening. The folds plunge to the southwest where they are incised and deflect the course of the Sacramento River. We will present preliminary geomorphic maps of fluvial terraces and drainage patterns associated with the folds constructed with standard field mapping techniques and aided by airborne LiDAR. It is anticipated that dating of deformed terrace strandlines with radiocarbon and OSL techniques holds the potential to quantify the lateral and vertical propagation rates of fold growth, as well as, constrain rates of regional contractional deformation.

  3. Insecticidal Effect of Chrysanthemum coronarium L. Flowers on the Pest Spodoptera littoralis Boisd and its Parasitoid Microplitis rufiventris Kok. with Identifying the Chemical Composition

    NASA Astrophysics Data System (ADS)

    Shonouda, Mourad L.; Osman, Salah; Salama, Osama; Ayoub, Amal

    The flower extract of Chrysanthemum coronarium L. and their fractions have shown insecticidal effect on the cotton leaf worm Spodoptera littoralis. The third instar larvae fed for two days on treated leaves were more susceptible to plant extracts and to their ethyl acetate and chloroform fractions. The active lowest concentration (5%) of the flower fractions showed no significant effect on the percent reduction of emerged adult parasitoids, Microplitis rufiventris Kok. GC/MS analysis revealed that the major constituents in ethyl acetate fraction were 3-dihydro-methylene-2- (3H) furanone (17.8%), jasmolin I (15.6%), carveol 1 (13.6%), phosphoric acid, tributyl ester (11.4%) and cinerin II (11.1%), while those of chloroform fraction were 5-hydroxy-3 methyl-1H-pyrazole (42.7%) and carveol 1(24.8%). The medicinal plant C. coronarium seems to be a promising plant for application in integrated pest management due to its safety to the surrounding environment.

  4. Imperfection works: Survival, transmission and persistence in the system of Heliothis virescens ascovirus 3h (HvAV-3h), Microplitis similis and Spodoptera exigua

    PubMed Central

    Li, Shun-Ji; Hopkins, Richard J.; Zhao, Yi-Pei; Zhang, Yun-Xuan; Hu, Jue; Chen, Xu-Yang; Xu, Zhi; Huang, Guo-Hua

    2016-01-01

    Ascoviruses are insect-specific large DNA viruses that mainly infect noctuid larvae, and are transmitted by parasitoids in the fields. Heliothis virescens ascovirus 3h (HvAV-3h) has been recently isolated from Spodoptera exigua, without parasitoid vector identified previously. Here we report that Microplitis similis, a solitary endoparasitoid wasp, could transmit HvAV-3h between S. exigua larvae in the laboratory. When the female parasitoid wasp acquired the virus and served as a vector, the period of virion viability on the ovipositor was 4.1 ± 1.4 days. Infected host larvae were still acceptable for egg laying by parasitoids, and the parasitoids thereafter transmitted virus to healthy hosts. Virus acquisition occurred only from donor hosts between 3 and 9 days post infection. The peak of virus acquisition (80.9 ± 6.3%) was found when M. similis wasps oviposited in larvae that had been inoculated with the virus 7 days previously. When virus infection of the host took place during the life cycle of the parasitoid wasp, it caused 1- to 4-day-old immature parasitoids death in the host, whilst a small proportion of 5- to 6-day-old and the majority of 7-day-old parasitoids larvae survived from the virus-infected hosts. Viral contamination did not reduce the life span or fecundity of female M. similis. PMID:26878829

  5. A microplate assay for the coupled transglycosylase-transpeptidase activity of the penicillin binding proteins; a vancomycin-neutralizing tripeptide combination prevents penicillin inhibition of peptidoglycan synthesis.

    PubMed

    Kumar, Vidya P; Basavannacharya, Chandrakala; de Sousa, Sunita M

    2014-07-18

    A microplate, scintillation proximity assay to measure the coupled transglycosylase-transpeptidase activity of the penicillin binding proteins in Escherichia coli membranes was developed. Membranes were incubated with the two peptidoglycan sugar precursors UDP-N-acetyl muramylpentapeptide (UDP-MurNAc(pp)) and UDP-[(3)H]N-acetylglucosamine in the presence of 40 μM vancomycin to allow in situ accumulation of lipid II. In a second step, vancomycin inhibition was relieved by addition of a tripeptide (Lys-D-ala-D-ala) or UDP-MurNAc(pp), resulting in conversion of lipid II to cross-linked peptidoglycan. Inhibitors of the transglycosylase or transpeptidase were added at step 2. Moenomycin, a transglycosylase inhibitor, had an IC50 of 8 nM. Vancomycin and nisin also inhibited the assay. Surprisingly, the transpeptidase inhibitors penicillin and ampicillin showed no inhibition. In a pathway assay of peptidoglycan synthesis, starting from the UDP linked sugar precursors, inhibition by penicillin was reversed by a 'neutral' combination of vancomycin plus tripeptide, suggesting an interaction thus far unreported. PMID:24944023

  6. Establishment of a microplate assay for flow cytometric assessment and it is use for the evaluation of age-related phenotypic changes in canine whole blood leukocytes.

    PubMed

    Reis, Alexandre B; Carneiro, Cláudia M; Carvalho, Maria das Graças; Teixeira-Carvalho, Andréa; Giunchetti, Rodolfo C; Mayrink, Wilson; Genaro, Odair; Corrêa-Oliveira, Rodrigo; Martins-Filho, Olindo A

    2005-02-10

    The effectiveness of flow cytometric assays for canine use is still requiring standardization. Despite several studies using purified mononuclear cells, no methodology or reference ranges are available for immunophenotyping of whole blood leukocytes (WBL). Fresh and pre-fixed WBL were used to identify cell-subsets, (Thy-1(+)/CD5(+)/CD4(+)/CD8(+)/CD21(+) and CD14(+)) and measure MHC-II, CD45RA/CD45RB expression. We described here an efficient method for fast quantification of canine-WBL, using pre-fix in a microplate assay, which allows long-term sample storage prior to phenotyping. Decreased percentage of CD5(+)-T-cells within the lymphocyte-gate and increased percentage of CD21(+)-B-cells were observed in young animals, which led to higher T/B cell ratios in middle-aged dogs. Lower numerical counts of Thy-1(+), CD4(+), CD8(+) and CD21(+) lymphocyte were observed when compared to young animals. In addition, we identified an age-related decline of MHC-II/CD45RA expression by lymphocytes. We proposed an improved method for phenotyping of canine peripheral blood mononuclear cells (PBMC) that has significant use for researchers and veterinary clinicians. The hematological changes of senescence previously identified on PBMC could be adequately reproduced on features identified by whole blood. Furthermore, this study supplies normal range references as baseline standards for clinical purposes, besides specific immunological parameters to monitor canine aging process. PMID:15621304

  7. Seismic hazard along the southern boundary of the Gônave microplate: block modelling of GPS velocities from Jamaica and nearby islands, northern Caribbean

    NASA Astrophysics Data System (ADS)

    Benford, B.; DeMets, C.; Tikoff, B.; Williams, P.; Brown, L.; Wiggins-Grandison, M.

    2012-07-01

    We use block modelling of GPS site velocities from Jamaica and nearby islands, including Hispaniola, to test alternative plate boundary geometries for deformation in Jamaica and estimate slip rates along the island's major fault zones. Relative to the Caribbean Plate, GPS sites in northern Jamaica move 6.0 ± 0.5 mm yr-1 to the WSW, constituting a lower bound on the motion of the Gônave microplate across its southern boundary in Jamaica. Obliquely convergent motion of all 30 GPS sites on and near Jamaica relative to the island's ˜E-W-trending strike-slip faults may be partitioned into 2.6 ± 0.6 mm yr-1 of ˜N-S shortening across submarine faults south of Jamaica and 5-6 mm yr-1 of E-W motion. Guided by geological and seismic information about the strikes and locations of faults in Jamaica, inverse block modelling of the regional GPS velocities rejects plate boundary configurations that presume either a narrow plate boundary in Jamaica or deformation concentrated across a restraining bend defined by the topographically high Blue Mountains of eastern Jamaica. The best-fitting models instead place most deformation on faults in central Jamaica. The 4-5 mm yr-1 slip rate we estimate for the Plantain Garden fault and Blue Mountain restraining bend of southeastern Jamaica implies significant seismic hazard for the nearby capital of Kingston.

  8. Imperfection works: Survival, transmission and persistence in the system of Heliothis virescens ascovirus 3h (HvAV-3h), Microplitis similis and Spodoptera exigua.

    PubMed

    Li, Shun-Ji; Hopkins, Richard J; Zhao, Yi-Pei; Zhang, Yun-Xuan; Hu, Jue; Chen, Xu-Yang; Xu, Zhi; Huang, Guo-Hua

    2016-01-01

    Ascoviruses are insect-specific large DNA viruses that mainly infect noctuid larvae, and are transmitted by parasitoids in the fields. Heliothis virescens ascovirus 3h (HvAV-3h) has been recently isolated from Spodoptera exigua, without parasitoid vector identified previously. Here we report that Microplitis similis, a solitary endoparasitoid wasp, could transmit HvAV-3h between S. exigua larvae in the laboratory. When the female parasitoid wasp acquired the virus and served as a vector, the period of virion viability on the ovipositor was 4.1 ± 1.4 days. Infected host larvae were still acceptable for egg laying by parasitoids, and the parasitoids thereafter transmitted virus to healthy hosts. Virus acquisition occurred only from donor hosts between 3 and 9 days post infection. The peak of virus acquisition (80.9 ± 6.3%) was found when M. similis wasps oviposited in larvae that had been inoculated with the virus 7 days previously. When virus infection of the host took place during the life cycle of the parasitoid wasp, it caused 1- to 4-day-old immature parasitoids death in the host, whilst a small proportion of 5- to 6-day-old and the majority of 7-day-old parasitoids larvae survived from the virus-infected hosts. Viral contamination did not reduce the life span or fecundity of female M. similis. PMID:26878829

  9. Listeria monocytogenes varies among strains to maintain intracellular pH homeostasis under stresses by different acids as analyzed by a high-throughput microplate-based fluorometry

    PubMed Central

    Cheng, Changyong; Yang, Yongchun; Dong, Zhimei; Wang, Xiaowen; Fang, Chun; Yang, Menghua; Sun, Jing; Xiao, Liya; Fang, Weihuan; Song, Houhui

    2015-01-01

    Listeria monocytogenes, a food-borne pathogen, has the capacity to maintain intracellular pH (pHi) homeostasis in acidic environments, but the underlying mechanisms remain elusive. Here, we report a simple microplate-based fluorescent method to determine pHi of listerial cells that were prelabeled with the fluorescent dye carboxyfluorescein diacetate N-succinimidyl ester and subjected to acid stress. We found that L. monocytogenes responds differently among strains toward organic and inorganic acids to maintain pHi homeostasis. The capacity of L. monocytogenes to maintain pHi at extracellular pH 4.5 (pHex) was compromised in the presence of acetic acid and lactic acid, but not by hydrochloric acid and citric acid. Organic acids exhibited more inhibitory effects than hydrochloric acid at certain pH conditions. Furthermore, the virulent stains L. monocytogenes EGDe, 850658 and 10403S was more resistant to acidic stress than the avirulent M7 which showed a defect in maintaining pHi homeostasis. Deletion of sigB, a stress-responsive alternative sigma factor from 10403S, markedly altered intracellular pHi homeostasis, and showed a significant growth and survival defect under acidic conditions. Thus, this work provides new insights into bacterial survival mechanism to acidic stresses. PMID:25667585

  10. Screening and monitoring microbial xenobiotics’ biodegradation by rapid, inexpensive and easy to perform microplate UV-absorbance measurements

    PubMed Central

    2014-01-01

    Background Evaluation of xenobiotics biodegradation potential, shown here for benzotriazoles (corrosion inhibitors) and sulfamethoxazole (sulfonamide antibiotic) by microbial communities and/or pure cultures normally requires time intensive and money consuming LC/GC methods that are, in case of laboratory setups, not always needed. Results The usage of high concentrations to apply a high selective pressure on the microbial communities/pure cultures in laboratory setups, a simple UV-absorbance measurement (UV-AM) was developed and validated for screening a large number of setups, requiring almost no preparation and significantly less time and money compared to LC/GC methods. This rapid and easy to use method was evaluated by comparing its measured values to LC-UV and GC-MS/MS results. Furthermore, its application for monitoring and screening unknown activated sludge communities (ASC) and mixed pure cultures has been tested and approved to detect biodegradation of benzotriazole (BTri), 4- and 5-tolyltriazole (4-TTri, 5-TTri) as well as SMX. Conclusions In laboratory setups, xenobiotics concentrations above 1.0 mg L-1 without any enrichment or preparation could be detected after optimization of the method. As UV-AM does not require much preparatory work and can be conducted in 96 or even 384 well plate formats, the number of possible parallel setups and screening efficiency was significantly increased while analytic and laboratory costs were reduced to a minimum. PMID:24558966

  11. 40Ar/39Ar geochronology of subaerial lava flows of Barren Island volcano and the deep crust beneath the Andaman Island Arc, Burma Microplate

    NASA Astrophysics Data System (ADS)

    Ray, Jyotiranjan S.; Pande, Kanchan; Bhutani, Rajneesh

    2015-06-01

    Little was known about the nature and origin of the deep crust beneath the Andaman Island Arc in spite of the fact that it formed part of the highly active Indonesian volcanic arc system, one of the important continental crust forming regions in Southeast Asia. This arc, formed as a result of subduction of the Indian Plate beneath the Burma Microplate (a sliver of the Eurasian Plate), contains only one active subaerial magmatic center, Barren Island volcano, whose evolutional timeline had remained uncertain. In this work, we present results of the first successful attempt to date crustal xenoliths and their host lava flows from the island, by incremental heating 40Ar/39Ar method, in an attempt to understand the evolutionary histories of the volcano and its basement. Based on concordant plateau and isochron ages, we establish that the oldest subaerial lava flows of the volcano are 1.58 ± 0.04 (2σ) Ma, and some of the plagioclase xenocrysts have been derived from crustal rocks of 106 ± 3 (2σ) Ma. Mineralogy (anorthite + Cr-rich diopside + minor olivine) and isotopic compositions (87Sr/86Sr < 0.7040; ɛNd > 7.0) of xenoliths not only indicate their derivation from a lower (oceanic) crustal olivine gabbro but also suggest a genetic relationship between the arc crust and the ophiolitic basement of the Andaman accretionary prism. We speculate that the basements of the forearc and volcanic arc of the Andaman subduction zone belong to a single continuous unit that was once attached to the western margin of the Eurasian Plate.

  12. Identifying active structures in the Kayak Island and Pamplona Zones: Implications for offshore tectonics of the Yakutat Microplate, Gulf of Alaska

    NASA Astrophysics Data System (ADS)

    Worthington, Lindsay L.; Gulick, Sean P. S.; Pavlis, Terry L.

    Within the northern Gulf of Alaska, the Yakutat (YAK) microplate obliquely collides with and subducts beneath the North American (NA) continent at near-Pacific plate velocities. We investigate the extent that thin-skinned deformation on offshore structures located within the western portion of the unsubducted YAK block accommodates YAK-NA convergence. We compare faulting and folding observed on high-resolution and basin-scale multichannel seismic (MCS) reflection data with earthquake locations and surface ruptures observed on high-resolution bathymetric data. Holocene sediments overlying the Kayak Island fault zone (KIZ), previously interpreted as a region of active contraction, are relatively flat-lying, suggesting that active convergence within the KIZ is waning. Seismic reflection profiles east of KIZ show up to ˜200 m of undisturbed sediments overlying older folds in the Bering Trough, indicating that this area has been tectonically inactive since at least the last ˜1.3 Ma. Farther east, MCS profiles image active deformation in surface sediments along the eastern edge of the Pamplona zone (PZ) fold-and-thrust belt, that are collocated with a concentration of earthquake events that continues southwest to Khitrov Ridge and onshore through Icy Bay. These observations suggest that during the late Quaternary offshore shallow deformation style changed from distributed across the western Yakutat block to localized at the eastern edge of the PZ with extrusion of sediments southwest through the Khitrov Ridge area to the Aleutian Trench. This shallow deformation is interpreted as deformation of an accretionary complex above a shallow decollement.

  13. Variation in the limit-of-detection of the ProSpecT Campylobacter microplate enzyme immunoassay in stools spiked with emerging Campylobacter species.

    PubMed

    Bojanić, Krunoslav; Midwinter, Anne Camilla; Marshall, Jonathan Craig; Rogers, Lynn Elizabeth; Biggs, Patrick Jon; Acke, Els

    2016-08-01

    Campylobacter enteritis in humans is primarily associated with C. jejuni/coli infection. The impact of other Campylobacter spp. is likely to be underestimated due to the bias of culture methods towards Campylobacter jejuni/coli diagnosis. Stool antigen tests are becoming increasingly popular and appear generally less species-specific. A review of independent studies of the ProSpecT® Campylobacter Microplate enzyme immunoassay (EIA) developed for C. jejuni/coli showed comparable diagnostic results to culture methods but the examination of non-jejuni/coli Campylobacter spp. was limited and the limit-of-detection (LOD), where reported, varied between studies. This study investigated LOD of EIA for Campylobacter upsaliensis, Campylobacter hyointestinalis and Campylobacter helveticus spiked in human stools. Multiple stools and Campylobacter isolates were used in three different concentrations (10(4)-10(9)CFU/ml) to reflect sample heterogeneity. All Campylobacter species evaluated were detectable by EIA. Multivariate analysis showed LOD varied between Campylobacter spp. and faecal consistency as fixed effects and individual faecal samples as random effects. EIA showed excellent performance in replicate testing for both within and between batches of reagents, in agreement between visual and spectrophotometric reading of results, and returned no discordance between the bacterial concentrations within independent dilution test runs (positive results with lower but not higher concentrations). This study shows how limitations in experimental procedures lead to an overestimation of consistency and uniformity of LOD for EIA that may not hold under routine use in diagnostic laboratories. Benefits and limitations for clinical practice and the influence on estimates of performance characteristics from detection of multiple Campylobacter spp. by EIA are discussed. PMID:27317896

  14. A microplate assay to assess chemical effects on RBL-2H3 mast cell degranulation: effects of triclosan without use of an organic solvent.

    PubMed

    Weatherly, Lisa M; Kennedy, Rachel H; Shim, Juyoung; Gosse, Julie A

    2013-01-01

    Mast cells play important roles in allergic disease and immune defense against parasites. Once activated (e.g. by an allergen), they degranulate, a process that results in the exocytosis of allergic mediators. Modulation of mast cell degranulation by drugs and toxicants may have positive or adverse effects on human health. Mast cell function has been dissected in detail with the use of rat basophilic leukemia mast cells (RBL-2H3), a widely accepted model of human mucosal mast cells(3-5). Mast cell granule component and the allergic mediator β-hexosaminidase, which is released linearly in tandem with histamine from mast cells(6), can easily and reliably be measured through reaction with a fluorogenic substrate, yielding measurable fluorescence intensity in a microplate assay that is amenable to high-throughput studies(1). Originally published by Naal et al.(1), we have adapted this degranulation assay for the screening of drugs and toxicants and demonstrate its use here. Triclosan is a broad-spectrum antibacterial agent that is present in many consumer products and has been found to be a therapeutic aid in human allergic skin disease(7-11), although the mechanism for this effect is unknown. Here we demonstrate an assay for the effect of triclosan on mast cell degranulation. We recently showed that triclosan strongly affects mast cell function(2). In an effort to avoid use of an organic solvent, triclosan is dissolved directly into aqueous buffer with heat and stirring, and resultant concentration is confirmed using UV-Vis spectrophotometry (using ε280 = 4,200 L/M/cm)(12). This protocol has the potential to be used with a variety of chemicals to determine their effects on mast cell degranulation, and more broadly, their allergic potential. PMID:24300285

  15. A Microplate Assay to Assess Chemical Effects on RBL-2H3 Mast Cell Degranulation: Effects of Triclosan without Use of an Organic Solvent

    PubMed Central

    Shim, Juyoung; Gosse, Julie A.

    2013-01-01

    Mast cells play important roles in allergic disease and immune defense against parasites. Once activated (e.g. by an allergen), they degranulate, a process that results in the exocytosis of allergic mediators. Modulation of mast cell degranulation by drugs and toxicants may have positive or adverse effects on human health. Mast cell function has been dissected in detail with the use of rat basophilic leukemia mast cells (RBL-2H3), a widely accepted model of human mucosal mast cells3-5. Mast cell granule component and the allergic mediator β-hexosaminidase, which is released linearly in tandem with histamine from mast cells6, can easily and reliably be measured through reaction with a fluorogenic substrate, yielding measurable fluorescence intensity in a microplate assay that is amenable to high-throughput studies1. Originally published by Naal et al.1, we have adapted this degranulation assay for the screening of drugs and toxicants and demonstrate its use here. Triclosan is a broad-spectrum antibacterial agent that is present in many consumer products and has been found to be a therapeutic aid in human allergic skin disease7-11, although the mechanism for this effect is unknown. Here we demonstrate an assay for the effect of triclosan on mast cell degranulation. We recently showed that triclosan strongly affects mast cell function2. In an effort to avoid use of an organic solvent, triclosan is dissolved directly into aqueous buffer with heat and stirring, and resultant concentration is confirmed using UV-Vis spectrophotometry (using ε280 = 4,200 L/M/cm)12. This protocol has the potential to be used with a variety of chemicals to determine their effects on mast cell degranulation, and more broadly, their allergic potential. PMID:24300285

  16. Transtensional Analyses of Fault Patterns and Strain Provinces of the Eastern California Shear Zone-Walker Lane on the Eastern Margin of the Sierra Nevada Microplate, California and Nevada

    NASA Astrophysics Data System (ADS)

    Taylor, T. R.; Dewey, J. F.

    2008-12-01

    Substantial work on the theory of transtensional strain at various scales has shown that transtension produces triaxial non-plane constrictional strains. At the plate boundary scale, fault geometries predicted by transtensional theory better explain observed fault patterns in the transtensional Eastern California shear zone-Walker Lane than 2D plane strain pure or simple shear solutions. Seemingly heterogeneous fault patterns and strain styles within structural provinces along the higher strain corridor adjacent to the eastern Sierra Nevada microplate margin are reconciled by non-plane strain evaluation, and in the context of triaxial, 3D strain partitioning characteristic of transtensional deformation. Among these structural provinces, which include the Honey Lake-Pyramid Lake region, the Lake Tahoe region, the Mono Lake-Long Valley region, Owens Valley, and the Coso region, all are found to be undergoing coaxially dominated transtension, except for the Mono Lake-Long Valley region which has possibly formed by noncoaxially dominated strain, and the northern Honey Lake-Pyramid Lake region, where the Sierra Nevada microplate boundary zone curves west toward the Mendocino triple junction. The local geometry of the transtensional zone boundary and the microplate transport direction determine the dominant type of non-plane strain. Fault orientations predicted by application of transtensional theory to identify instantaneous strain axes are consistent with those observed in each structural province, and comparatively, are not adequately explained by plane strain kinematic models. The orientations and amount of shortening and elongation of the finite strain ellipsoid axes for each province indicate smaller amounts of shortening, elongation and rotation of axes, and overall less ellipticity, in the coaxially dominated strain areas, and greater shortening, elongation, and rotation, and overall greater ellipticity, in the noncoaxially dominated areas. Exceptions are the

  17. Immobilization of saccharides and peptides on 96-well microtiter plates coated with methyl vinyl ether-maleic anhydride copolymer.

    PubMed

    Satoh, A; Kojima, K; Koyama, T; Ogawa, H; Matsumoto, I

    1998-06-15

    We have previously reported a method to immobilize protein ligands on microtiter plates coated with methyl vinyl ether-maleic anhydride copolymer (MMAC) [Isosaki, K., et al. (1992) J. Chromatogr. 597, 123-128]. In this study, we improved the MMAC method to efficiently immobilize not only small ligands such as peptides and oligosaccharides, which could not be efficiently immobilized previously, but also heparin via its reducing end. Amino and hydrazino groups were introduced to MMAC-coated microtiter plate wells by coupling to acid anhydride groups of MMAC with 1,6-hexamethylenediamine and adipic acid dihydrazide, respectively. The amino groups introduced were allowed to react with peptides by use of divalent cross-linkers. Hydrazino groups were allowed to react with formyl groups of saccharides by reductive amination. Peptides and oligosaccharides were immobilized in a dose-dependent manner by these methods. In the case of the angiotensin peptide thus immobilized, the detection limit by monoclonal antibodies was as low as 0.1-1 fmol peptide per well. Application of 20-200 nmol oligosaccharides to the well was sufficient to immobilize and subsequently detect lectins. Furthermore, heparin immobilized on the hydrazinocoated wells was successfully used for the binding assay of annexin IV. PMID:9648659

  18. Geodetic Constraints on the Rigidity and Eastern Boundary of the Sierra Nevada Micro-Plate, from Mohawk Valley to Southern Walker Lane

    NASA Astrophysics Data System (ADS)

    Kreemer, C. W.; Hammond, W. C.; Blewitt, G.

    2009-12-01

    The Sierra Nevada - Great Valley (SNGV) micro-plate has long been recognized as a tectonically rigid, though mobile, entity within the Pacific - North America plate boundary zone. The motion of the SNGV relative to stable North America (and the Colorado Plateau) provides the kinematic boundary condition for, and perhaps drives, the deformation in the Basin and Range Province (BRP) and Walker Lane. In the north the motion of the SNGV is aligned with the Mohawk Valley fault zone, which could have a slip rate of over a few mm/yr. The crest of the Sierras marks the SNGV’s eastern edge, but the obliquity between orientation of this boundary and the block’s motion implies an expected increase in rangefront-normal extension from the northern to southern Walker Lane. We use new GPS data from the EarthScope Plate Boundary Observatory (PBO) and our own semi-continuous MAGNET network to revisit the following questions: 1) Do the data still support rigidity of the SNGV?; 2) How far east does the rigidity extend and how does this relate to SNGV lithology?; 3) How does the direction of SNGV motion relate to the strike of its eastern margin and observed strain partitioning (and its along strike variation) in the Walker Lane?; and 4) How is SNGV-BRP motion accommodated between the Walker Lane and the Cascadia forearc? We analyze data from all the available continuous GPS sites in the greater SNGV region, including new data from PBO, as well as data from MAGNET. All data are processed with the GIPSY-OASIS II precise point positioning software using recently reprocessed orbits from JPL's IGS Analysis Center. The processing includes satellite and station antenna calibrations and all data have the phase ambiguities fixed using the Ambizap algorithm. Positions are estimated in our custom-made North America reference frame in which continental-scale common-mode errors are removed. Velocities and uncertainties are estimated using the CATS software in which we assuming an error model

  19. Concept Formation.

    ERIC Educational Resources Information Center

    Vaidya, Narendera

    This document, published in India by the Regional College of Education, deals with 13 subjects: the tough context (thinking), definitions of concept, functions of concept, the process of concept formation, discriminant learning, mediation process, second signalling system, factors affecting concept formation, studies in concept formation, the…

  20. Optimizing Two-Color Semiconductor Nanocrystal Immunoassays in Single Well Microtiter Plate Formats

    PubMed Central

    Sapsford, Kim E.; Spindel, Samantha; Jennings, Travis; Tao, Guoliang; Triulzi, Robert C.; Algar, W. Russ; Medintz, Igor L.

    2011-01-01

    The simultaneous detection of two analytes, chicken IgY (IgG) and Staphylococcal enterotoxin B (SEB), in the single well of a 96-well plate is demonstrated using luminescent semiconductor quantum dot nanocrystal (NC) tracers. The NC-labeled antibodies were prepared via sulfhydryl-reactive chemistry using a facile protocol that took <3 h. Dose response curves for each target were evaluated in a single immunoassay format and compared to Cy5, a fluorophore commonly used in fluorescent immunoassays, and found to be equivalent. Immunoassays were then performed in a duplex format, demonstrating multiplex detection in a single well with limits of detection equivalent to the single assay format: 9.8 ng/mL chicken IgG and 7.8 ng/mL SEB. PMID:22164051

  1. [Study on virulence factors associated with biofilm formation and phylogenetic groupings in Escherichia coli strains isolated from patients with cystitis].

    PubMed

    Tiba, Monique Ribeiro; Nogueira, Gustavo Prado; Leite, Domingos da Silva

    2009-01-01

    Escherichia coli samples isolated from female patients with cystitis were characterized with regard to the presence of virulence factors associated with biofilm formation and phylogenetic groupings. Polymerase chain reaction results demonstrated that all the samples were positive for the gene fimH (type 1 fimbriae), 91 for fliC (flagellins), 50 for papC (P fimbriae), 44 for kpsMTII (capsules) and 36 for flu (antigen 43). The results from assays to quantify the biofilm formation demonstrated that 44 samples produced biofilm on polystyrene microplates and 56 samples produced weak or no biofilm. We also confirmed that Escherichia coli samples were present in phylogenetic groups B2 and D. PMID:19287937

  2. Formative Assessment

    ERIC Educational Resources Information Center

    Technology & Learning, 2005

    2005-01-01

    In today's climate of high-stakes testing and accountability, educators are challenged to continuously monitor student progress to ensure achievement. This article details how formative assessment helps educators meet this challenge and to ensure achievement. Formative assessment can influence learning and support achievement, allowing teachers…

  3. Effect of Lactobacillus species on Streptococcus mutans biofilm formation.

    PubMed

    Ahmed, Ayaz; Dachang, Wu; Lei, Zhou; Jianjun, Liu; Juanjuan, Qiu; Yi, Xin

    2014-09-01

    Streptococcus mutans is the primary pathogen responsible for initiating dental caries and decay. The presence of sucrose, stimulates S. mutans to produce insoluble glucans to form oral biofilm also known as dental plaque to initiate caries lesion. The GtfB and LuxS genes of S. mutans are responsible for formation and maturation of biofilm. Lactobacillus species as probiotic can reduces the count of S. mutans. In this study effect of different Lactobacillus species against the formation of S. mutans biofilm was observed. Growing biofilm in the presence of sucrose was detected using 96 well microtiter plate crystal violet assay and biofilm formation by S. mutans in the presence of Lactobacillus was detected. Gene expression of biofilm forming genes (GtfB and LuxS) was quantified through Real-time PCR. All strains of Lactobacillus potently reduced the formation of S. mutans biofilm whereas Lactobacillus acidophilus reduced the genetic expression by 60-80%. Therefore, probiotic Lactobacillus species can be used as an alternative instead of antibiotics to decrease the chance of dental caries by reducing the count of S. mutans and their gene expression to maintain good oral health. PMID:25176247

  4. Effect of Honey on Streptococcus mutans Growth and Biofilm Formation

    PubMed Central

    Li, Mingyun

    2012-01-01

    Because of the tradition of using honey as an antimicrobial medicament, we investigated the effect of natural honey (NH) on Streptococcus mutans growth, viability, and biofilm formation compared to that of an artificial honey (AH). AH contained the sugars at the concentrations reported for NH. NH and AH concentrations were obtained by serial dilution with tryptic soy broth (TSB). Several concentrations of NH and AH were tested for inhibition of bacterial growth, viability, and biofilm formation after inoculation with S. mutans UA159 in 96-well microtiter plates to obtain absorbance and CFU values. Overall, NH supported significantly less (P < 0.05) bacterial growth than AH at 25 and 12.5% concentrations. At 50 and 25% concentrations, both honey groups provided significantly less bacterial growth and biofilm formation than the TSB control. For bacterial viability, the results for all honey concentrations except 50% NH were not significantly different from those for the TSB control. NH was able to decrease the maximum velocity of S. mutans growth compared to AH. In summary, NH demonstrated more inhibition of bacterial growth, viability, and biofilm formation than AH. This study highlights the potential antibacterial properties of NH and could suggest that the antimicrobial mechanism of NH is not solely due to its high sugar content. PMID:22038612

  5. Determination of colchicine and O-demethylated metabolites in decomposed skeletal tissues by microwave assisted extraction, microplate solid phase extraction and ultra-high performance liquid chromatography (MAE-MPSPE-UHPLC).

    PubMed

    Watterson, J H; Imfeld, A B; Cornthwaite, H C

    2014-06-01

    Microwave assisted extraction (MAE) followed by microplate solid phase extraction (MPSPE) coupled with ultra high performance liquid chromatography (UHPLC) for the semi-quantitative determination of colchicine, 3-demethyl colchicine and 2-demethyl colchicine in postmortem rat bone is described. Rats (n=4) received 50mg/kg colchicine (i.p), and euthanized by CO2 asphyxiation. Remains decomposed to skeleton outdoors and vertebral bones were collected cleaned, and ground to a fine powder. Powdered bone underwent MAE using methanol in a closed microwave system, followed by MPSPE and analysis using UHPLC-PDA. MAE analyte stability was assessed and found to be stable for at least 60 min irradiation time. The majority (>95%) of each analyte was recovered after 15 min. The MPSPE-UHPLC method was linear between 10 and 2,000 ng/mL, with coefficients of variation <20% in triplicate analysis, with a limit of detection of 10 ng/mL for each of the three analytes. Following MAE for 30 min (80°C, 1200W), MPSPE-UHPLC analysis of vertebral bone of colchicine-exposed rats detected colchicine (1.8-4.1 μg/g), 3-demethyl colchicine (0.77-1.8 μg/g) and 2-demethyl colchicine (0.43-0.80 μg/g) in all samples assayed. PMID:24799069

  6. Latest Precambrian to Early Cambrian U-Pb zircon ages of augen gneisses from Calabria (Italy), with inference to the Alboran microplate in the evolution of the peri-Gondwana terranes

    NASA Astrophysics Data System (ADS)

    Micheletti, Francesca; Barbey, Pierre; Fornelli, Annamaria; Piccarreta, Giuseppe; Deloule, Etienne

    2007-10-01

    In situ U-Pb dating of zircons from five samples of Calabrian augen gneisses shows that their protoliths are Latest Precambrian to Early Cambrian in age (562 ± 15, 547 ± 7, 540 ± 4, 539 ± 16 and 526 ± 10 Ma), and contain Archaean (3.1 Ga), Palaeoproterozoic (1.7-2.4 Ga) and Neoproterozoic (0.6-0.9 Ga) inheritance. Geochemical signature of augen gneisses is typical of high-K calc-alkaline post-collisional magmatism. Their Sr-Nd isotopic compositions [0.7093 < (87Sr/86Sr)i < 0.7139; -3.2 < ɛNd(t) < -5.4; 1.5 < T DM < 1.7 Ga] indicate the involvement of a crustal component in significant proportions. The Calabrian augen gneisses have, therefore, to be distinguished from the orthogneisses of Sardinia and northern Algeria, and from the porphyroids of Sicily, which are Middle Ordovician. By contrast, the Calabrian augen gneisses show a close similarity to the Pan-African post-collisional granitoids of the northern edge of the West African craton (e.g. the Moroccan Anti-Atlas). This suggests a peri-Gondwana origin and corroborates previous palaeogeodynamic reconstructions attributing the Alboran microplate to the northern margin of the West African craton.

  7. Electroantennogram (EAG) responses of Microplitis croceipes and Cotesia marginiventris and their lepidopteran hosts to a wide array of odor stimuli: correlation between EAG response and degree of host specificity?

    PubMed

    Ngumbi, Esther; Chen, Li; Fadamiro, Henry

    2010-09-01

    In order to test whether the electroantennogram (EAG) response spectrum of an insect correlates to its degree of host specificity, we recorded EAG responses of two parasitoid species with different degrees of host specificity, Microplitis croceipes (specialist) and Cotesia marginiventris (generalist), to a wide array of odor stimuli including compounds representing green leaf volatiles (GLVs), herbivore-induced plant volatiles (HIPV), ecologically irrelevant (not used by the parasitoid species and their hosts for host location) plant volatiles, and host-specific odor stimuli (host sex pheromones, and extracts of host caterpillar body and frass). We also tested the EAG responses of female moths of the caterpillar hosts of the parasitoids, Heliothis virescens and Spodoptera exigua, to some of the odor stimuli. We hypothesized that the specialist parasitoid will have a narrower EAG response spectrum than the generalist, and that the two lepidopteran species, which are similar in their host plant use, will show similar EAG response spectra to plant volatiles. As predicted, the specialist parasitoid showed greater EAG responses than the generalist to host-specific odor and one HIPV (cis-3-hexenyl butyrate), whereas the generalist showed relatively greater EAG responses to the GLVs and unrelated plant volatiles. We detected no differences in the EAG responses of H. virescens and S. exigua to any of the tested odor. PMID:20371248

  8. Galaxy formation

    SciTech Connect

    Silk, J.

    1984-11-01

    Implications of the isotropy of the cosmic microwave background on large and small angular scales for galaxy formation are reviewed. In primeval adiabatic fluctuations, a universe dominated by cold, weakly interacting nonbaryonic matter, e.g., the massive photino is postulated. A possible signature of photino annihilation in our galactic halo involves production of cosmic ray antiprotons. If the density is near its closure value, it is necessary to invoke a biasing mechanism for suppressing galaxy formation throughout most of the universe in order to reconcile the dark matter density with the lower astronomical determinations of the mean cosmological density. A mechanism utilizing the onset of primordial massive star formation to strip gaseous protogalaxies is described. Only the densest, early collapsing systems form luminous galaxies. (ESA)

  9. Comet formation

    NASA Astrophysics Data System (ADS)

    Blum, J.

    2014-07-01

    There has been vast progress in our understanding of planetesimal formation over the past decades, owing to a number of laboratory experiments as well as to refined models of dust and ice agglomeration in protoplanetary disks. Coagulation rapidly forms cm-sized ''pebbles'' by direct sticking in collisions at low velocities (Güttler et al. 2010; Zsom et al. 2010). For the further growth, two model approaches are currently being discussed: (1) Local concentration of pebbles in nebular instabilities until gravitational instability occurs (Johansen et al. 2007). (2) A competition between fragmentation and mass transfer in collisions among the dusty bodies, in which a few ''lucky winners'' make it to planetesimal sizes (Windmark et al. 2012a,b; Garaud et al. 2013). Predictions of the physical properties of the resulting bodies in both models allow a distinction of the two formation scenarios of planetesimals. In particular, the tensile strength (i.e, the inner cohesion) of the planetesimals differ widely between the two models (Skorov & Blum 2012; Blum et al. 2014). While model (1) predicts tensile strengths on the order of ˜ 1 Pa, model (2) results in rather compactified dusty bodies with tensile strengths in the kPa regime. If comets are km-sized survivors of the planetesimal-formation era, they should in principle hold the secret of their formation process. Water ice is the prime volatile responsible for the activity of comets. Thermophysical models of the heat and mass transport close to the comet-nucleus surface predict water-ice sublimation temperatures that relate to maximum sublimation pressures well below the kPa regime predicted for formation scenario (2). Model (1), however, is in agreement with the observed dust and gas activity of comets. Thus, a formation scenario for cometesimals involving gravitational instability is favored (Blum et al. 2014).

  10. Planet Formation

    NASA Astrophysics Data System (ADS)

    Klahr, Hubert; Brandner, Wolfgang

    2011-02-01

    1. Historical notes on planet formation Bodenheimer; 2. The formation and evolution of planetary systems Bouwman et al.; 3. Destruction of protoplanetary disks by photoevaporation Richling, Hollenbach and Yorke; 4. Turbulence in protoplanetary accretion disks Klahr, Rozyczka, Dziourkevitch, Wunsch and Johansen; 5. The origin of solids in the early solar system Trieloff and Palme; 6. Experiments on planetesimal formation Wurm and Blum; 7. Dust coagulation in protoplanetary disks Henning, Dullemond, Wolf and Dominik; 8. The accretion of giant planet cores Thommes and Duncan; 9. Planetary transits: direct vision of extrasolar planets Lecavelier des Etangs and Vidal-Madjar; 10. The core accretion - gas capture model Hubickyj; 11. Properties of exoplanets Marcy, Fischer, Butler and Vogt; 12. Giant planet formation: theories meet observations Boss; 13. From hot Jupiters to hot Neptures … and below Lovis, Mayor and Udry; 14. Disk-planet interaction and migration Masset and Kley; 15. The Brown Dwarf - planet relation Bate; 16. From astronomy to astrobiology Brandner; 17. Overview and prospective Lin.

  11. In vitro prevention of Pseudomonas aeruginosa early biofilm formation with antibiotics used in cystic fibrosis patients.

    PubMed

    Fernández-Olmos, Ana; García-Castillo, María; Maiz, Luis; Lamas, Adelaida; Baquero, Fernando; Cantón, Rafael

    2012-08-01

    The ability of antibiotics used in bronchopulmonary infections in cystic fibrosis (CF) patients to prevent Pseudomonas aeruginosa early biofilm formation was studied using a biofilm microtitre assay with 57 non-mucoid P. aeruginosa isolates (44 first colonisers and 13 recovered during the initial intermittent colonisation stage) obtained from 35 CF patients. Minimum biofilm inhibitory concentrations (BICs) of levofloxacin, ciprofloxacin, imipenem, ceftazidime, tobramycin, colistin and azithromycin were determined by placing a peg lid with a formed biofilm onto microplates containing antibiotics. A modification of this protocol consisting of antibiotic challenge during biofilm formation was implemented in order to determine the biofilm prevention concentration (BPC), i.e. the minimum concentration able to prevent biofilm formation. The lowest BPCs were for fluoroquinolones, tobramycin and colistin and the highest for ceftazidime and imipenem. The former antibiotics had BPCs identical to or only slightly higher than their minimum inhibitory concentrations (MICs) determined by standard Clinical and Laboratory Standards Institute (CLSI) microdilution and were also active on formed biofilms as reflected by their low BIC values. In contrast, ceftazidime and imipenem were less effective for prevention of biofilm formation and on formed biofilms. In conclusion, the new BPC parameter determined in non-mucoid P. aeruginosa isolates recovered during early colonisation stages in CF patients supports early aggressive antimicrobial treatment guidelines in first P. aeruginosa-colonised CF patients. PMID:22727530

  12. Tissue Plasminogen Activator Coating on Implant Surfaces Reduces Staphylococcus aureus Biofilm Formation

    PubMed Central

    Na, Manli; Jarneborn, Anders; Jacobsson, Gunnar; Peetermans, Marijke; Verhamme, Peter

    2015-01-01

    Staphylococcus aureus biofilm infections of indwelling medical devices are a major medical challenge because of their high prevalence and antibiotic resistance. As fibrin plays an important role in S. aureus biofilm formation, we hypothesize that coating of the implant surface with fibrinolytic agents can be used as a new method of antibiofilm prophylaxis. The effect of tissue plasminogen activator (tPA) coating on S. aureus biofilm formation was tested with in vitro microplate biofilm assays and an in vivo mouse model of biofilm infection. tPA coating efficiently inhibited biofilm formation by various S. aureus strains. The effect was dependent on plasminogen activation by tPA, leading to subsequent local fibrin cleavage. A tPA coating on implant surfaces prevented both early adhesion and later biomass accumulation. Furthermore, tPA coating increased the susceptibility of biofilm infections to antibiotics. In vivo, significantly fewer bacteria were detected on the surfaces of implants coated with tPA than on control implants from mice treated with cloxacillin. Fibrinolytic coatings (e.g., with tPA) reduce S. aureus biofilm formation both in vitro and in vivo, suggesting a novel way to prevent bacterial biofilm infections of indwelling medical devices. PMID:26519394

  13. Formation testers

    SciTech Connect

    Brieger, E.

    1980-07-01

    A description is given of a method for use in obtaining multiple pressure tests of an earth formation traversed by a well bore by use of a sidewall fluid sampler well tool which has a fluid pressure sampling chamber in the well tool in open fluid communication with a pad sealing means, comprising the steps of: for one selected level in a well bore, moving a pad sealing means on the well tool into engagement with the wall of a well bore and isolating a wall segment of the earth formation; after the pad sealing means engges the wall segment of the earth formation, generating a hydraulic pressure in the well tool and applying said hydraulic pressure to said fluid pressure sampling chamber for increasing the volume of said fluid pressure sampling chamber thereby to dray a fluid sample from the earth formation engaged by the pad sealing means into the fluid pressure sampling chamber, sensing the pressure of said fluid sample as it is drawn into the fluid pressure sampling chamber while the volume of the sampling chamber is being increased, relieving the hydraulic pressure in the well tool with respect to said fluid pressur sampling chamber for decreasing the volume of said fluid pressure sampling chamber thereby to contact the sampling chamber to dischrge the fluid sample through the pad sealing means; retracting the sealing pad means and, after retrction of sealing pad means from engagement from the wall of the well bore, moving the well tool to a second location at another level in the well bore and, at the second location, repeating the steps of the method performed at the one selected level for obtaining another fluid sample and pressure sensing at said second location.

  14. Amphiplex Formation

    NASA Astrophysics Data System (ADS)

    Petersen, Shannon; Laaser, Jennifer; Lodge, Timothy

    2015-03-01

    Polymer-micelle complexes are currently under heavy investigation due to their potential applications in targeted drug delivery and gene therapy, yet the dynamics of the complex formation is still relatively unstudied. By varying the ratios of poly(styrene sulfonate) chains and cationic poly(dimethylaminoethyl methacrylate)-b-poly(styrene) micelles and the ionic strength of the system, we created a variety of complex configurations of different sizes and charges. The complexes were characterized dynamic light scattering and zeta potential measurements which provided information regarding the hydrodynamic radius, distribution of sizes, and effective charge.

  15. The mode of biofilm formation on smooth surfaces by Campylobacter jejuni.

    PubMed

    Moe, Kyaw Kyaw; Mimura, Junichiro; Ohnishi, Takahiro; Wake, Tomoya; Yamazaki, Wataru; Nakai, Masaaki; Misawa, Naoaki

    2010-04-01

    Many microorganisms produce extracellular polymers referred to collectively as "slime" or glycocalyx, and form biofilms on solid surfaces in natural ecosystems. Campylobacter jejuni, one of the most important foodborne pathogens, also has the ability to form biofilm on stainless steel, glass, or polyvinyl chloride in vitro. However, the issue of biofilm formation by Campylobacter species has not been extensively examined. The present study was performed to examine the mode of adhesion of C. jejuni to a smooth surface. When bacterial suspensions in Brucella broth were incubated in microplate wells with a glass coverslip, microcolonies 0.5~2 mm in diameter were formed on the coverslip within 2 hr from the start of incubation. These microcolonies gradually grew and formed a biofilm of net-like connections within 6 hr. Transmission electron microscopy indicated that massive amounts of extracellular material masked the cell surface, and this material bound ruthenium red, suggesting the presence of a polysaccharide moiety. Scanning electron microscopy indicated that the flagella acted as bridges, forming net-like connections between the organisms. To determine the genes associated with biofilm formation, aflagellate (flaA(-)) and flagellate but non-motile (motA(-)) mutants were constructed from strain 81-176 by natural transformation-mediated allelic exchange. The flaA(-) and motA(-) mutants did not form the biofilm exhibited by the wild-type strain. These findings suggest that flagella-mediated motility as well as flagella is required for biofilm formation in vitro. PMID:20009353

  16. Habit formation.

    PubMed

    Smith, Kyle S; Graybiel, Ann M

    2016-03-01

    Habits, both good ones and bad ones, are pervasive in animal behavior. Important frameworks have been developed to understand habits through psychological and neurobiological studies. This work has given us a rich understanding of brain networks that promote habits, and has also helped us to understand what constitutes a habitual behavior as opposed to a behavior that is more flexible and prospective. Mounting evidence from studies using neural recording methods suggests that habit formation is not a simple process. We review this evidence and take the position that habits could be sculpted from multiple dissociable changes in neural activity. These changes occur across multiple brain regions and even within single brain regions. This strategy of classifying components of a habit based on different brain signals provides a potentially useful new way to conceive of disorders that involve overly fixed behaviors as arising from different potential dysfunctions within the brain's habit network. PMID:27069378

  17. Habit formation

    PubMed Central

    Smith, Kyle S.; Graybiel, Ann M.

    2016-01-01

    Habits, both good ones and bad ones, are pervasive in animal behavior. Important frameworks have been developed to understand habits through psychological and neurobiological studies. This work has given us a rich understanding of brain networks that promote habits, and has also helped us to understand what constitutes a habitual behavior as opposed to a behavior that is more flexible and prospective. Mounting evidence from studies using neural recording methods suggests that habit formation is not a simple process. We review this evidence and take the position that habits could be sculpted from multiple dissociable changes in neural activity. These changes occur across multiple brain regions and even within single brain regions. This strategy of classifying components of a habit based on different brain signals provides a potentially useful new way to conceive of disorders that involve overly fixed behaviors as arising from different potential dysfunctions within the brain's habit network. PMID:27069378

  18. Barrier Formation

    PubMed Central

    Lyaruu, D.M.; Medina, J.F.; Sarvide, S.; Bervoets, T.J.M.; Everts, V.; DenBesten, P.; Smith, C.E.; Bronckers, A.L.J.J.

    2014-01-01

    Enamel fluorosis is an irreversible structural enamel defect following exposure to supraoptimal levels of fluoride during amelogenesis. We hypothesized that fluorosis is associated with excess release of protons during formation of hypermineralized lines in the mineralizing enamel matrix. We tested this concept by analyzing fluorotic enamel defects in wild-type mice and mice deficient in anion exchanger-2a,b (Ae2a,b), a transmembrane protein in maturation ameloblasts that exchanges extracellular Cl− for bicarbonate. Defects were more pronounced in fluorotic Ae2a,b−/− mice than in fluorotic heterozygous or wild-type mice. Phenotypes included a hypermineralized surface, extensive subsurface hypomineralization, and multiple hypermineralized lines in deeper enamel. Mineral content decreased in all fluoride-exposed and Ae2a,b−/− mice and was strongly correlated with Cl−. Exposure of enamel surfaces underlying maturation-stage ameloblasts to pH indicator dyes suggested the presence of diffusion barriers in fluorotic enamel. These results support the concept that fluoride stimulates hypermineralization at the mineralization front. This causes increased release of protons, which ameloblasts respond to by secreting more bicarbonates at the expense of Cl− levels in enamel. The fluoride-induced hypermineralized lines may form barriers that impede diffusion of proteins and mineral ions into the subsurface layers, thereby delaying biomineralization and causing retention of enamel matrix proteins. PMID:24170372

  19. Pattern Formation

    NASA Astrophysics Data System (ADS)

    Hoyle, Rebecca

    2006-03-01

    From the stripes of a zebra and the spots on a leopard's back to the ripples on a sandy beach or desert dune, regular patterns arise everywhere in nature. The appearance and evolution of these phenomena has been a focus of recent research activity across several disciplines. This book provides an introduction to the range of mathematical theory and methods used to analyse and explain these often intricate and beautiful patterns. Bringing together several different approaches, from group theoretic methods to envelope equations and theory of patterns in large-aspect ratio-systems, the book also provides insight behind the selection of one pattern over another. Suitable as an upper-undergraduate textbook for mathematics students or as a fascinating, engaging, and fully illustrated resource for readers in physics and biology, Rebecca Hoyle's book, using a non-partisan approach, unifies a range of techniques used by active researchers in this growing field. Accessible description of the mathematical theory behind fascinating pattern formation in areas such as biology, physics and materials science Collects recent research for the first time in an upper level textbook Features a number of exercises - with solutions online - and worked examples

  20. The BioFilm Ring Test: a Rapid Method for Routine Analysis of Pseudomonas aeruginosa Biofilm Formation Kinetics

    PubMed Central

    Olivares, Elodie; Badel-Berchoux, Stéphanie; Provot, Christian; Jaulhac, Benoît; Prévost, Gilles; Bernardi, Thierry

    2015-01-01

    Currently, few techniques are available for the evaluation of bacterial biofilm adhesion. These detection tools generally require time for culture and/or arduous handling steps. In this work, the BioFilm Ring Test (BRT), a new technology, was used to estimate the biofilm formation kinetics of 25 strains of Pseudomonas aeruginosa, isolated from the sputum of cystic fibrosis (CF) patients. The principle of the new assay is based on the mobility measurement of magnetic microbeads mixed with a bacterial suspension in a polystyrene microplate. If free to move under the magnetic action, particles gather to a visible central spot in the well bottom. Therefore, the absence of spot formation in the plate reflects the bead immobilization by a biofilm in formation. The BRT device allowed us to classify the bacterial strains into three general adhesion profiles. Group 1 consists of bacteria, which are able to form a solid biofilm in <2 h. Group 2 comprises the strains that progressively set up a biofilm during 24 h. Lastly, group 3 includes the strains that stay in a planktonic form. The grouping of our strains did not differ according to culture conditions, i.e., the use of different sets of beads or culture media. The BRT is shown to be an informative tool for the characterization of biofilm-forming bacteria. Various application perspectives may be investigated for this device, such as the addition of antibiotics to the bacterial suspension to select which would have the ability to inhibit the biofilm formation. PMID:26719437

  1. The BioFilm Ring Test: a Rapid Method for Routine Analysis of Pseudomonas aeruginosa Biofilm Formation Kinetics.

    PubMed

    Olivares, Elodie; Badel-Berchoux, Stéphanie; Provot, Christian; Jaulhac, Benoît; Prévost, Gilles; Bernardi, Thierry; Jehl, François

    2016-03-01

    Currently, few techniques are available for the evaluation of bacterial biofilm adhesion. These detection tools generally require time for culture and/or arduous handling steps. In this work, the BioFilm Ring Test (BRT), a new technology, was used to estimate the biofilm formation kinetics of 25 strains of Pseudomonas aeruginosa, isolated from the sputum of cystic fibrosis (CF) patients. The principle of the new assay is based on the mobility measurement of magnetic microbeads mixed with a bacterial suspension in a polystyrene microplate. If free to move under the magnetic action, particles gather to a visible central spot in the well bottom. Therefore, the absence of spot formation in the plate reflects the bead immobilization by a biofilm in formation. The BRT device allowed us to classify the bacterial strains into three general adhesion profiles. Group 1 consists of bacteria, which are able to form a solid biofilm in <2 h. Group 2 comprises the strains that progressively set up a biofilm during 24 h. Lastly, group 3 includes the strains that stay in a planktonic form. The grouping of our strains did not differ according to culture conditions, i.e., the use of different sets of beads or culture media. The BRT is shown to be an informative tool for the characterization of biofilm-forming bacteria. Various application perspectives may be investigated for this device, such as the addition of antibiotics to the bacterial suspension to select which would have the ability to inhibit the biofilm formation. PMID:26719437

  2. Microfabricated glucose biosensor for culture well operation.

    PubMed

    Pemberton, R M; Cox, T; Tuffin, R; Sage, I; Drago, G A; Biddle, N; Griffiths, J; Pittson, R; Johnson, G; Xu, J; Jackson, S K; Kenna, G; Luxton, R; Hart, J P

    2013-04-15

    A water-based carbon screen-printing ink formulation, containing the redox mediator cobalt phthalocyanine (CoPC) and the enzyme glucose oxidase (GOx), was investigated for its suitability to fabricate glucose microbiosensors in a 96-well microplate format: (1) the biosensor ink was dip-coated onto a platinum (Pt) wire electrode, leading to satisfactory amperometric performance; (2) the ink was deposited onto the surface of a series of Pt microelectrodes (10-500 μm diameter) fabricated on a silicon substrate using MEMS (microelectromechanical systems) microfabrication techniques: capillary deposition proved to be successful; a Pt microdisc electrode of ≥100 μm was required for optimum biosensor performance; (3) MEMS processing was used to fabricate suitably sized metal (Pt) tracks and pads onto a silicon 96 well format base chip, and the glucose biosensor ink was screen-printed onto these pads to create glucose microbiosensors. When formed into microwells, using a 340 μl volume of buffer, the microbiosensors produced steady-state amperometric responses which showed linearity up to 5 mM glucose (CV=6% for n=5 biosensors). When coated, using an optimised protocol, with collagen in order to aid cell adhesion, the biosensors continued to show satisfactory performance in culture medium (linear range to 2 mM, dynamic range to 7 mM, CV=5.7% for n=4 biosensors). Finally, the operation of these collagen-coated microbiosensors, in 5-well 96-well format microwells, was tested using a 5-channel multipotentiostat. A relationship between amperometric response due to glucose, and cell number in the microwells, was observed. These results indicate that microphotolithography and screen-printing techniques can be combined successfully to produce microbiosensors capable of monitoring glucose metabolism in 96 well format cell cultures. The potential application areas for these microbiosensors are discussed. PMID:23265827

  3. A simplified 96-well method for the estimation of phenolic acids and antioxidant activity from eggplant pulp extracts using UV spectral scan data

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eggplant fruit is ranked amongst the top ten vegetables in terms of oxygen radical absorbance capacity due to its high phenolic acid content. The main objective of this study was to determine if a simple UV spectral analysis method can be used as a screening tool to estimate the amount of phenolic ...

  4. Tectonic Processes Along the Southeastern Margin of Alaska - The Neogene Sedimentary Record: Yakataga Formation, St. Elias Mountains

    NASA Astrophysics Data System (ADS)

    Witmer, J. W.; Ridgway, K. D.; Brennan, P. R.; Arnaud, E.; Pavlis, T.

    2008-12-01

    Neogene collision of the Yakutat microplate with the southern Alaskan continental margin is associated with extreme rates of exhumation and erosion of the St. Elias Mountains. The exhumation and the concurrent development of temperate glaciers are recorded in the ~5000 m of sedimentary strata of the Yakataga Formation. We present new data from measured stratigraphic sections that document along-strike and temporal changes within the Yakataga Formation along this collisional margin during Miocene to Pleistocene time. In the eastern part of our study area, the Yakataga Formation consists of lenticular sandstone and conglomerate facies associated with fan-delta depositional environments that are overlain by thick-bedded glaciomarine strata. These strata grade to finer-grained sandstone and convoluted mudstone typical of marine shelf environments in the central part of our study area. Along strike in the westernmost part of our study area the Yakataga Formation is interpreted to be laterally equivalent to Neogene strata of the Redwood Formation. These strata include thick-bedded, macrofossil-rich sandstone, well-rounded conglomerate, and thin-bedded mudstone facies that are characteristic of nearshore and shelf depositional environments. These sediments were likely sourced by fluvial systems along the continental margin that served as the backstop for Neogene collision. Preliminary compositional data also suggest that the Redwood Formation was derived from a different source than the Yakataga Formation. Along-strike changes in structural configuration of the Yakataga Formation are also observed. In the easternmost part of our study area adjacent to the Dangerous River zone (DRZ), a possible remnant strike-slip fault system, unconformities between the Yakataga Formation and underlying strata require erosion of 1000s of meters of missing Eocene-Miocene strata. We interpret this part of the mountain range to have undergone the greatest amount of Neogene exhumation. In the

  5. Relationship of biofilm formation and different virulence genes in uropathogenic Escherichia coli isolates from Northwest Iran

    PubMed Central

    Fattahi, Sargol; Kafil, Hossein Samadi; Nahai, Mohammad Reza; Asgharzadeh, Mohammad; Nori, Roghaya; Aghazadeh, Mohammad

    2015-01-01

    Background and objectives: The Escherichia coli (E. coli) bacterium is one of the main causative agents of urinary tract infections (UTI) worldwide. The ability of this bacterium to form biofilms on medical devices such as catheters plays an important role in the development of UTI. The aim of the present study was to investigate the possible relationship between virulence factors and biofilm formation of E. coli isolates responsible for urinary tract infection. Materials and methods: A total of 100 E. coli isolates isolated from patients with UTI were collected and characterized by routine bacteriological methods. In vitro biofilm formation by these isolates was determined using the 96-well microtiter-plate test, and the presence of fimA, papC, and hly virulence genes was examined by PCR assay. Data analysis was performed using SPSS 16.0 software. Results: From 100 E. coli isolates isolated from UTIs, 92% were shown to be biofilm positive. The genes papC, fimA, and hly were detected in 43%, 94% and 26% of isolates, respectively. Biofilm formation in isolates that expressed papC, fimA, and hly genes was 100%, 93%, and 100%, respectively. A significant relationship was found between presence of the papC gene and biofilm formation in E. coli isolates isolated from UTI (P<0.01), but there was no statistically significant correlation between presence of fimA and hly genes with biofilm formation (P<0.072, P<0.104). Conclusion: Results showed that fimA and hly genes do not seem to be necessary or sufficient for the production of biofilm in E. coli, but the presence of papC correlates with increased biofilm formation of urinary tract isolates. Overall, the presence of fimA, papC, and hly virulence genes coincides with in vitro biofilm formation in uropathogenic E. coli isolates. PMID:26213679

  6. Nitric oxide regulates neutrophil migration through microparticle formation.

    PubMed

    Nolan, Sarah; Dixon, Rachel; Norman, Keith; Hellewell, Paul; Ridger, Victoria

    2008-01-01

    The role of nitric oxide (NO) in regulating neutrophil migration has been investigated. Human neutrophil migration to interleukin (IL)-8 (1 nmol/L) was measured after a 1-hour incubation using a 96-well chemotaxis plate assay. The NO synthase inhibitor N(G)-nitro-l-arginine methyl ester (L-NAME) significantly (P < 0.001) enhanced IL-8-induced migration by up to 45%. Anti-CD18 significantly (P < 0.001) inhibited both IL-8-induced and L-NAME enhanced migration. Antibodies to L-selectin or PSGL-1 had no effect on IL-8-induced migration but prevented the increased migration to IL-8 induced by L-NAME. L-NAME induced generation of neutrophil-derived microparticles that was significantly (P < 0.01) greater than untreated neutrophils or D-NAME. This microparticle formation was dependent on calpain activity and superoxide production. Only microparticles from L-NAME and not untreated or D-NAME-treated neutrophils induced a significant (P < 0.01) increase in IL-8-induced migration and transendothelial migration. Pretreatment of microparticles with antibodies to L-selectin (DREG-200) or PSGL-1 (PL-1) significantly (P < 0.001) inhibited this effect. The ability of L-NAME-induced microparticles to enhance migration was found to be dependent on the number of microparticles produced and not an increase in microparticle surface L-selectin or PSGL-1 expression. These data show that NO can modulate neutrophil migration by regulating microparticle formation. PMID:18079439

  7. A novel two-component response regulator links rpf with biofilm formation and virulence of Xanthomonas axonopodis pv. citri.

    PubMed

    Huang, Tzu-Pi; Lu, Kuan-Min; Chen, Yu-Hsuan

    2013-01-01

    Citrus bacterial canker caused by Xanthomonas axonopodis pv. citri is a serious disease that impacts citrus production worldwide, and X. axonopodis pv. citri is listed as a quarantine pest in certain countries. Biofilm formation is important for the successful development of a pathogenic relationship between various bacteria and their host(s). To understand the mechanisms of biofilm formation by X. axonopodis pv. citri strain XW19, the strain was subjected to transposon mutagenesis. One mutant with a mutation in a two-component response regulator gene that was deficient in biofilm formation on a polystyrene microplate was selected for further study. The protein was designated as BfdR for biofilm formation defective regulator. BfdR from strain XW19 shares 100% amino acid sequence identity with XAC1284 of X. axonopodis pv. citri strain 306 and 30-100% identity with two-component response regulators in various pathogens and environmental microorganisms. The bfdR mutant strain exhibited significantly decreased biofilm formation on the leaf surfaces of Mexican lime compared with the wild type strain. The bfdR mutant was also compromised in its ability to cause canker lesions. The wild-type phenotype was restored by providing pbfdR in trans in the bfdR mutant. Our data indicated that BfdR did not regulate the production of virulence-related extracellular enzymes including amylase, lipase, protease, and lecithinase or the expression of hrpG, rfbC, and katE; however, BfdR controlled the expression of rpfF in XVM2 medium, which mimics cytoplasmic fluids in planta. In conclusion, biofilm formation on leaf surfaces of citrus is important for canker development in X. axonopodis pv. citri XW19. The process is controlled by the two-component response regulator BfdR via regulation of rpfF, which is required for the biosynthesis of a diffusible signal factor. PMID:23626857

  8. Bisbibenzyls, a New Type of Antifungal Agent, Inhibit Morphogenesis Switch and Biofilm Formation through Upregulation of DPP3 in Candida albicans

    PubMed Central

    Zhang, Li; Chang, Wenqiang; Sun, Bin; Groh, Matthias; Speicher, Andreas; Lou, Hongxiang

    2011-01-01

    The yeast-to-hypha transition plays a crucial role in the pathogenesis of C. albicans. Farnesol, a quorum sensing molecule (QSM) secreted by the fungal itself, could prevent the formation of hyphae and subsequently lead to the defect of biofilm formation. The DPP3, encoding phosphatase, is a key gene in regulating farnesol synthesis. In this study, we screened 24 bisbibenzyls and 2 bibenzyls that were isolated from bryophytes or chemically synthesized by using CLSI method for antifungal effect. Seven bisbibenzyls were found to have antifungal effects with IC80 less than 32 µg/ml, and among them, plagiochin F, isoriccardin C and BS-34 were found to inhibit the hyphae and biofilm formation of C. albicans in a dose-dependent manner. To uncover the underlying relationship between morphogenesis switch and QSM formation, we measured the farnesol production by HPLC-MS and quantified Dpp3 expression by detecting the fluorescent intensity of green fluorescent protein tagged strain using Confocal Laser Scanning microscopy and Multifunction Microplate Reader. The DPP3 transcripts were determined by real-time PCR. The data indicated that the bisbibenzyls exerted antifungal effects through stimulating the synthesis of farnesol via upregulation of Dpp3, suggesting a potential antifungal application of bisbibenzyls. In addition, our assay provides a novel, visual and convenient method to measure active compounds against morphogenesis switch. PMID:22174935

  9. Star formation - An overview

    NASA Technical Reports Server (NTRS)

    Evans, N. J., II

    1985-01-01

    Methods for studying star formation are reviewed. Stellar clusters and associations, as well as field stars, provide a fossil record of the star formation process. Regions of current star formation provide a series of snapshots of different epochs of star formation. A simplified picture of individual star formation as it was envisioned in the late 1970s is contrasted with the results of recent observations, in particular the outflow phenomenon.

  10. Formate Formation and Formate Conversion in Biological Fuels Production

    PubMed Central

    Crable, Bryan R.; Plugge, Caroline M.; McInerney, Michael J.; Stams, Alfons J. M.

    2011-01-01

    Biomethanation is a mature technology for fuel production. Fourth generation biofuels research will focus on sequestering CO2 and providing carbon-neutral or carbon-negative strategies to cope with dwindling fossil fuel supplies and environmental impact. Formate is an important intermediate in the methanogenic breakdown of complex organic material and serves as an important precursor for biological fuels production in the form of methane, hydrogen, and potentially methanol. Formate is produced by either CoA-dependent cleavage of pyruvate or enzymatic reduction of CO2 in an NADH- or ferredoxin-dependent manner. Formate is consumed through oxidation to CO2 and H2 or can be further reduced via the Wood-Ljungdahl pathway for carbon fixation or industrially for the production of methanol. Here, we review the enzymes involved in the interconversion of formate and discuss potential applications for biofuels production. PMID:21687599

  11. Inhibitory effect of gels loaded with a low concentration of antibiotics against biofilm formation by Enterococcus faecalis and Porphyromonas gingivalis.

    PubMed

    A Algarni, Amnah; H Yassen, Ghaeth; L Gregory, Richard

    2015-09-01

    We explored longitudinally the inhibitory effect of gels loaded with 1 mg/mL modified triple antibiotic paste (MTAP) or double antibiotic paste (DAP) against biofilm formation by Enterococcus faecalis and Porphyromonas gingivalis. Methylcellulose-based antibiotic gels of MTAP (ciprofloxacin, metronidazole and clindamycin) and DAP (ciprofloxacin and metronidazole) were prepared at a concentration of 1 mg/mL. Individually cultured E. faecalis and P. gingivalis bacterial suspensions were treated with MTAP, DAP, or placebo (vehicle only) gels at different dilutions and allowed to grow in 96-well microtiter plates. Untreated bacterial suspensions served as a negative control. Crystal violet assays were used to evaluate biofilm formation after 48 h. The ability of the gels to inhibit biofilm formation was determined immediately, and at 1 month and 3 months after the gels had been prepared. Data were analyzed using a mixed-model ANOVA. The MTAP and DAP gels significantly reduced biofilm formation by both bacterial species at all time points, regardless of the tested dilution. No-significant differences in biofilm-inhibitory effects between MTAP and DAP gels were observed at the majority of the tested dilutions through various time points. Gels loaded with 1 mg/mL MTAP and DAP demonstrated a significant antibiofilm effect against E.faecalis and P. gingivalis. PMID:26369485

  12. Biofilms of vaginal Lactobacillus reuteri CRL 1324 and Lactobacillus rhamnosus CRL 1332: kinetics of formation and matrix characterization.

    PubMed

    Leccese Terraf, María Cecilia; Juárez Tomás, María Silvina; Rault, Lucie; Le Loir, Yves; Even, Sergine; Nader-Macías, María Elena Fátima

    2016-09-01

    Adhesion and biofilm formation are strain properties that reportedly contribute to the permanence of lactobacilli in the human vagina. The kinetics of biofilm formation and the chemical nature of the biofilm matrix formed by Lactobacillus reuteri CRL (Centro de Referencia para Lactobacilos Culture Collection) 1324 and Lactobacillus rhamnosus CRL 1332, vaginal beneficial strains, were evaluated in this work. Crystal violet-stained microplate assay and techniques of epifluorescence, electron and confocal microscopy were applied. The highest density and complexity of biofilms of both vaginal lactobacilli were observed at 72 h of incubation. Protease, proteinase K, α-chymotrypsin and trypsin treatments efficiently detached L. reuteri CRL 1324 biofilm that was also partially affected by α-amylase. However, L. rhamnosus CRL 1332 biofilm was slightly affected by protease, proteinase K and α-amylase. Confocal microscopy revealed greater amount of polysaccharides in L. rhamnosus CRL 1332 biofilm matrix than in L. reuteri CRL 1324 biofilm matrix. The results indicate that proteins are one of the main components of the L. reuteri CRL 1324 biofilm, while the biofilm matrix of L. rhamnosus CRL 1332 is composed of carbohydrates and proteins. The results obtained support the knowledge, understanding and characterization of two biofilm-forming vaginal Lactobacillus strains. PMID:27146055

  13. High throughput kinetic Vibrio fischeri bioluminescence inhibition assay for study of toxic effects of nanoparticles.

    PubMed

    Mortimer, M; Kasemets, K; Heinlaan, M; Kurvet, I; Kahru, A

    2008-08-01

    Despite of the growing production and use of nanoparticles (NPs) in various applications, current regulations, including EC new chemical policy REACH, fail to address the environmental, health, and safety risks posed by NPs. This paper shows that kinetic Vibrio fischeri luminescence inhibition test--Flash Assay--that up to now was mainly used for toxicity analysis of solid and colored environmental samples (e.g. sediments, soil suspensions), is a powerful tool for screening the toxic properties of NPs. To demonstrate that Flash Assay (initially designed for a tube luminometer) can also be adapted to a microplate format for high throughput toxicity screening of NPs, altogether 11 chemicals were comparatively analyzed. The studied chemicals included bulk and nanosized CuO and ZnO, polyethylenimine (PEI) and polyamidoamine dendrimer generations 2 and 5 (PAMAM G2 and G5). The results showed that EC50 values of 30-min Flash Assay in tube and microplate formats were practically similar and correlated very well (log-logR2=0.98), classifying all analyzed chemicals, except nano CuO (that was more toxic in cuvette format), analogously when compared to the risk phrases of the EC Directive 93/67/EEC for ranking toxicity of chemicals for aquatic organisms. The 30-min EC50 values of nanoscale organic cationic polymers (PEI and dendrimers) ranged from 215 to 775 mg/l. Thirty-minute EC50 values of metal oxides varied largely, ranging from approximately 4 mg/l (bulk and nano ZnO) to approximately 100 mg/l (nano CuO) and approximately 4000 mg/l (bulk CuO). Thus, considering an excellent correlation between both formats, 96-well microplate Flash Assay can be successfully used for high throughput evaluation of harmful properties of chemicals (including organic and inorganic NPs) to bacteria. PMID:18400463

  14. Effects of short-chain fatty acids on Actinomyces naeslundii biofilm formation.

    PubMed

    Yoneda, S; Kawarai, T; Narisawa, N; Tuna, E B; Sato, N; Tsugane, T; Saeki, Y; Ochiai, K; Senpuku, H

    2013-10-01

    Actinomyces naeslundii is an early colonizer and has important roles in the development of the oral biofilm. Short-chain fatty acids (SCFA) are secreted extracellularly as a product of metabolism by gram-negative anaerobes, e.g. Porphyromonas gingivalis and Fusobacterium nucleatum; and the SCFA may affect biofilm development with interaction between A. naeslundii and gram-negative bacteria. Our aim was to investigate the effects of SCFA on biofilm formation by A. naeslundii and to determine the mechanism. We used the biofilm formation assay in 96-well microtiter plates in tryptic soy broth without dextrose and with 0.25% sucrose using safranin stain of the biofilm monitoring 492 nm absorbance. To determine the mechanism by SCFA, the production of chaperones and stress-response proteins (GrpE and GroEL) in biofilm formation was examined using Western blot fluorescence activity with GrpE and GroEL antibodies. Adding butyric acid (6.25 mm) 0, 6 and 10 h after beginning culture significantly increased biofilm formation by A. naeslundii, and upregulation was observed at 16 h. Upregulation was also observed using appropriate concentrations of other SCFA. In the upregulated biofilm, production of GrpE and GroEL was higher where membrane-damaged or dead cells were also observed. The upregulated biofilm was significantly reduced by addition of anti-GroEL antibody. The data suggest biofilm formation by A. naeslundii was upregulated dependent on the production of stress proteins, and addition of SCFA increased membrane-damaged or dead cells. Production of GroEL may physically play an important role in biofilm development. PMID:23731652

  15. Structure Formation in Astrophysics

    NASA Astrophysics Data System (ADS)

    Chabrier, Gilles

    2009-01-01

    Part I. Physical Processes and Numerical Methods Common to Structure Formations in Astrophysics: 1. The physics of turbulence E. Levêque; 2. The numerical simulation of turbulence W. Schmidt; 3. Numerical methods for radiation magnetohydrodynamics in astrophysics R. Klein and J. Stone; 4. The role of jets in the formation of planets, stars, and galaxies R. Banerjee, R. Pudritz and R. Ouyed; 5. Advanced numerical methods in astrophysical fluid dynamics A. Hujeirat and F. Heitsch; Part II. Structure and Star Formation in the Primordial Universe: 6. New frontiers in cosmology and galaxy formation challenges for the future R. Ellis and J. Silk; 7. Galaxy formation physics T. Abel, G. Bryan and R. Teyssier; 8. First stars formation, evolution, feedback effects V. Bromm, A. Ferrara and A. Heger; Part III. Contemporary Star and Brown Dwarf Formation: a) Cloud Formation and Fragmentation: 9. Diffuse interstellar medium and the formation of molecular clouds P. Hennebelle, M. Mac Low and E. Vazquez-Semadeni; 10. The formation of distributed and clustered stars in molecular clouds T. Megeath, Z. -Y. Li and A. Nordlund; b) Core Fragmentation and Star Formation: 11. The formation and evolution of prestellar cores P. André, S. Basu and S. Inutsuka; 12. Models for the formation of massive stars; Part IV. Protoplanetary Disks and Planet Formation M. Krumholz and I. Bonnell: 13. Observational properties of disks and young stellar objects G. Duchêne, F. Ménard, J. Muzzerolle and S. Mohanty; 14. Structure and dynamics of protoplanetary disks C. Dullemond, R. Durisen and J. Papaloizou; 15. Planet formation and evolution theory and observation Y. Alibert, I. Baraffe, W. Benz, G. Laughlin and S. Udry; 16. Planet formation assembling the puzzle G. Wurm and T. Guillot; Part V. Summary: 17. Open issues in small- and large-scale structure formation R. Klessen and M. Mac Low; 18. Final word E. Salpeter.

  16. Structure Formation in Astrophysics

    NASA Astrophysics Data System (ADS)

    Chabrier, Gilles

    2011-02-01

    Part I. Physical Processes and Numerical Methods Common to Structure Formations in Astrophysics: 1. The physics of turbulence E. Levêque; 2. The numerical simulation of turbulence W. Schmidt; 3. Numerical methods for radiation magnetohydrodynamics in astrophysics R. Klein and J. Stone; 4. The role of jets in the formation of planets, stars, and galaxies R. Banerjee, R. Pudritz and R. Ouyed; 5. Advanced numerical methods in astrophysical fluid dynamics A. Hujeirat and F. Heitsch; Part II. Structure and Star Formation in the Primordial Universe: 6. New frontiers in cosmology and galaxy formation challenges for the future R. Ellis and J. Silk; 7. Galaxy formation physics T. Abel, G. Bryan and R. Teyssier; 8. First stars formation, evolution, feedback effects V. Bromm, A. Ferrara and A. Heger; Part III. Contemporary Star and Brown Dwarf Formation: a) Cloud Formation and Fragmentation: 9. Diffuse interstellar medium and the formation of molecular clouds P. Hennebelle, M. Mac Low and E. Vazquez-Semadeni; 10. The formation of distributed and clustered stars in molecular clouds T. Megeath, Z. -Y. Li and A. Nordlund; b) Core Fragmentation and Star Formation: 11. The formation and evolution of prestellar cores P. André, S. Basu and S. Inutsuka; 12. Models for the formation of massive stars; Part IV. Protoplanetary Disks and Planet Formation M. Krumholz and I. Bonnell: 13. Observational properties of disks and young stellar objects G. Duchêne, F. Ménard, J. Muzzerolle and S. Mohanty; 14. Structure and dynamics of protoplanetary disks C. Dullemond, R. Durisen and J. Papaloizou; 15. Planet formation and evolution theory and observation Y. Alibert, I. Baraffe, W. Benz, G. Laughlin and S. Udry; 16. Planet formation assembling the puzzle G. Wurm and T. Guillot; Part V. Summary: 17. Open issues in small- and large-scale structure formation R. Klessen and M. Mac Low; 18. Final word E. Salpeter.

  17. Star Formation in Galaxies

    NASA Technical Reports Server (NTRS)

    1987-01-01

    Topics addressed include: star formation; galactic infrared emission; molecular clouds; OB star luminosity; dust grains; IRAS observations; galactic disks; stellar formation in Magellanic clouds; irregular galaxies; spiral galaxies; starbursts; morphology of galactic centers; and far-infrared observations.

  18. The Format Dilemma.

    ERIC Educational Resources Information Center

    Oder, Norman

    2002-01-01

    Reports results of a survey of public libraries that investigated trends in audiovisual materials. Highlights include format issues; audiobooks; media budgets for various formats; video collections; DVDs; circulation; collection sizes; music CDs; and future possibilities. (LRW)

  19. Simple Ontology Format (SOFT)

    SciTech Connect

    Sorokine, Alexandre

    2011-10-01

    Simple Ontology Format (SOFT) library and file format specification provides a set of simple tools for developing and maintaining ontologies. The library, implemented as a perl module, supports parsing and verification of the files in SOFt format, operations with ontologies (adding, removing, or filtering of entities), and converting of ontologies into other formats. SOFT allows users to quickly create ontologies using only a basic text editor, verify it, and portray it in a graph layout system using customized styles.

  20. Sensitive high-throughput screening for the detection of reducing sugars.

    PubMed

    Mellitzer, Andrea; Glieder, Anton; Weis, Roland; Reisinger, Christoph; Flicker, Karlheinz

    2012-01-01

    The exploitation of renewable resources for the production of biofuels relies on efficient processes for the enzymatic hydrolysis of lignocellulosic materials. The development of enzymes and strains for these processes requires reliable and fast activity-based screening assays. Additionally, these assays are also required to operate on the microscale and on the high-throughput level. Herein, we report the development of a highly sensitive reducing-sugar assay in a 96-well microplate screening format. The assay is based on the formation of osazones from reducing sugars and para-hydroxybenzoic acid hydrazide. By using this sensitive assay, the enzyme loads and conversion times during lignocellulose hydrolysis can be reduced, thus allowing higher throughput. The assay is about five times more sensitive than the widely applied dinitrosalicylic acid based assay and can reliably detect reducing sugars down to 10 μM. The assay-specific variation over one microplate was determined for three different lignocellulolytic enzymes and ranges from 2 to 8%. Furthermore, the assay was combined with a microscale cultivation procedure for the activity-based screening of Pichia pastoris strains expressing functional Thermomyces lanuginosus xylanase A, Trichoderma reesei β-mannanase, or T. reesei cellobiohydrolase 2. PMID:21538898

  1. Medical image file formats.

    PubMed

    Larobina, Michele; Murino, Loredana

    2014-04-01

    Image file format is often a confusing aspect for someone wishing to process medical images. This article presents a demystifying overview of the major file formats currently used in medical imaging: Analyze, Neuroimaging Informatics Technology Initiative (Nifti), Minc, and Digital Imaging and Communications in Medicine (Dicom). Concepts common to all file formats, such as pixel depth, photometric interpretation, metadata, and pixel data, are first presented. Then, the characteristics and strengths of the various formats are discussed. The review concludes with some predictive considerations about the future trends in medical image file formats. PMID:24338090

  2. Quorum quenching bacteria isolated from the sludge of a wastewater treatment plant and their application for controlling biofilm formation.

    PubMed

    Kim, A-Leum; Park, Son-Young; Lee, Chi-Ho; Lee, Chung-Hak; Lee, Jung-Kee

    2014-11-28

    Bacteria recognize changes in their population density by sensing the concentration of signal molecules, N-acyl-homoserine lactones (AHLs). AHL-mediated quorum sensing (QS) plays a key role in biofilm formation, so the interference of QS, referred to as quorum quenching (QQ), has received a great deal of attention. A QQ strategy can be applied to membrane bioreactors (MBRs) for advanced wastewater treatment to control biofouling. To isolate QQ bacteria that can inhibit biofilm formation, we isolated diverse AHL-degrading bacteria from a laboratory-scale MBR and sludge from real wastewater treatment plants. A total of 225 AHLdegrading bacteria were isolated from the sludge sample by enrichment culture. To identify the enzyme responsible for AHL degradation in QQ bacteria, AHL-degrading activities were analyzed using cell-free lysate, culture supernatant, and whole cells. Afipia sp. and Acinetobacter sp. strains produced the intracellular QQ enzyme, whereas Pseudomonas sp. and Micrococcus sp. produced the extracellular QQ enzyme that was most likely to produce AHLacylase. AHL-degrading activity was observed in whole-cell assay with the Microbacterium sp. and Rhodococcus sp. strains. There has been no report for AHL-degrading capability in the case of Streptococcus sp. and Afipia sp. strains. Finally, inhibition of biofilm formation by isolated QQ bacteria or enzymes was observed on glass slides and 96-well microtiter plates using crystal violet staining. QQ strains or enzymes not only inhibited initial biofilm development but also reduced established biofilms. PMID:25112313

  3. Calcium Increases Xylella fastidiosa Surface Attachment, Biofilm Formation, and Twitching Motility

    PubMed Central

    Cruz, Luisa F.; Cobine, Paul A.

    2012-01-01

    Xylella fastidiosa is a plant-pathogenic bacterium that forms biofilms inside xylem vessels, a process thought to be influenced by the chemical composition of xylem sap. In this work, the effect of calcium on the production of X. fastidiosa biofilm and movement was analyzed under in vitro conditions. After a dose-response study with 96-well plates using eight metals, the strongest increase of biofilm formation was observed when medium was supplemented with at least 1.0 mM CaCl2. The removal of Ca by extracellular (EGTA, 1.5 mM) and intracellular [1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid acetoxymethyl ester (BAPTA/AM), 75 μM] chelators reduced biofilm formation without compromising planktonic growth. The concentration of Ca influenced the force of adhesion to the substrate, biofilm thickness, cell-to-cell aggregation, and twitching motility, as shown by assays with microfluidic chambers and other assays. The effect of Ca on attachment was lost when cells were treated with tetracycline, suggesting that Ca has a metabolic or regulatory role in cell adhesion. A double mutant (fimA pilO) lacking type I and type IV pili did not improve biofilm formation or attachment when Ca was added to the medium, while single mutants of type I (fimA) or type IV (pilB) pili formed more biofilm under conditions of higher Ca concentrations. The concentration of Ca in the medium did not significantly influence the levels of exopolysaccharide produced. Our findings indicate that the role of Ca in biofilm formation may be related to the initial surface and cell-to-cell attachment and colonization stages of biofilm establishment, which rely on critical functions by fimbrial structures. PMID:22194297

  4. Star Formation for Predictive Primordial Galaxy Formation

    NASA Astrophysics Data System (ADS)

    Milosavljević, Miloš; Safranek-Shrader, Chalence

    The elegance of inflationary cosmology and cosmological perturbation theory ends with the formation of the first stars and galaxies, the initial sources of light that launched the phenomenologically rich process of cosmic reionization. Here we review the current understanding of early star formation, emphasizing unsolved problems and technical challenges. We begin with the first generation of stars to form after the Big Bang and trace how they influenced subsequent star formation. The onset of chemical enrichment coincided with a sharp increase in the overall physical complexity of star forming systems. Ab-initio computational treatments are just now entering the domain of the predictive and are establishing contact with local observations of the relics of this ancient epoch.

  5. Tropical cyclone formation

    SciTech Connect

    Montgomery, M.T.; Farrell, B.F. )

    1993-01-15

    The physics of tropical cyclone formation is not well understood, and more is known about the mature hurricane than the formative mechanisms that produce it. It is believed part of the reason for this can be traced to insufficient upper-level atmospheric data. Recent observations suggest that tropical cyclones are initiated by asymmetric interactions associated with migratory upper-level potential vorticity disturbances and low-level disturbances. Favored theories of cyclones formation, however, focus on internal processes associated with cumulus convection and/or air-sea interaction. This work focuses on external mechanisms of cyclone formation and, using both a two- and three-dimensional moist geostrophic momentum model, investigates the role of upper-level potential vorticity disturbances on the formation process. A conceptual model of tropical cyclone formation is proposed, and implications of the theory are discussed. 71 refs., 5 figs., 1 tab.

  6. Data format translation routines

    SciTech Connect

    Burris, R.D.

    1981-02-01

    To enable the effective connection of several dissimilar computers into a network, modification of the data being passed from one computer to another may become necessary. This document describes a package of routines which permit the translation of data in PDP-8 formats to PDP-11 or DECsystem-10 formats or from PDP-11 format to DECsystem-10 format. Additional routines are described which permit the effective use of the translation routines in the environment of the Fusion Energy Division (FED) network and the Elmo Bumpy Torus (EBT) data base.

  7. Simple Ontology Format (SOFT)

    2011-10-01

    Simple Ontology Format (SOFT) library and file format specification provides a set of simple tools for developing and maintaining ontologies. The library, implemented as a perl module, supports parsing and verification of the files in SOFt format, operations with ontologies (adding, removing, or filtering of entities), and converting of ontologies into other formats. SOFT allows users to quickly create ontologies using only a basic text editor, verify it, and portray it in a graph layoutmore » system using customized styles.« less

  8. METHODS OF EXPLORING METABOLIC STRUCTURE AND TAXONOMIC DIVERSITY RELATIONSHIPS BETWEEN BACTERIOPLANKTON AND PHYTOPLANKTON IN SALT MARSH TIDAL CREEKS

    EPA Science Inventory

    Bacterial metabolic diversity and phytoplankton community diversity were examined in eight shallow tidal creeks over a two-year period (1997-1998) within North Inlet estuary, South Carolina. The BIOLOG 96-well microplate method was used to assess metabolic diversity of bacteria, ...

  9. Optimization of Spore and Antifungal Lipopeptide Production during the Solid State Fermentation of Bacillus subtilis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacillus subtilis strain TrigoCor 1448 was grown on wheat middlings in 0.5-liter solid state fermentation (SSF) bioreactors for the production of an antifungal biological control agent. Total antifungal activity was quantified using a 96-well microplate bioassay against the plant pathogen Fusarium ...

  10. EPA worst case water microcosms for testing phage biocontrol of Salmonella

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A microplate method was developed as a tool to test phages for their ability to control Salmonella in aqueous environments. The method used EPA (U.S. Environmental Protection Agency) worst case water (WCW) in 96-well plates. The WCW provided a consistent and relatively simple defined turbid aqueou...

  11. GLOBAL STAR FORMATION REVISITED

    SciTech Connect

    Silk, Joseph; Norman, Colin E-mail: norman@stsci.edu

    2009-07-20

    A general treatment of disk star formation is developed from a dissipative multiphase model, with the dominant dissipation due to cloud collisions. The Schmidt-Kennicutt (SK) law emerges naturally for star-forming disks and starbursts. We predict that there should be an inverse correlation between Tully-Fisher law and SK law residuals. The model is extended to include a multiphase treatment of supernova feedback that leads to a turbulent pressure-regulated generalization of the star formation law and is applicable to gas-rich starbursts. Enhanced pressure, as expected in merger-induced star formation, enhances star formation efficiency. An upper limit is derived for the disk star formation rate in starbursts that depends on the ratio of global ISM to cloud pressures. We extend these considerations to the case where the interstellar gas pressure in the inner galaxy is dominated by outflows from a central active galactic nucleus (AGN). During massive spheroid formation, AGN-driven winds trigger star formation, resulting in enhanced supernova feedback and outflows. The outflows are comparable to the AGN-boosted star formation rate and saturate in the super-Eddington limit. Downsizing of both SMBH and spheroids is a consequence of AGN-driven positive feedback. Bondi accretion feeds the central black hole with a specific accretion rate that is proportional to the black hole mass. AGN-enhanced star formation is mediated by turbulent pressure and relates spheroid star formation rate to black hole accretion rate. The relation between black hole mass and spheroid velocity dispersion has a coefficient (Salpeter time to gas consumption time ratio) that provides an arrow of time. Highly efficient, AGN-boosted star formation can occur at high redshift.

  12. Rapid LC-MS Drug Metabolite Profiling Using Microsomal Enzyme Bioreactors in a Parallel Processing Format

    PubMed Central

    Bajrami, Besnik; Zhao, Linlin; Schenkman, John B.; Rusling, James F.

    2009-01-01

    Silica nanoparticle bioreactors featuring thin films of enzymes and polyions were utilized in a novel high-throughput 96-well plate format for drug metabolism profiling. The utility of the approach was illustrated by investigating the metabolism of the drugs diclofenac (DCF), troglitazone (TGZ) and raloxifene, for which we observed known metabolic oxidation and bioconjugation pathways and turnover rates. A broad range of enzymes was included by utilizing human liver (HLM), rat liver (RLM) and bicistronic human-cyt P450 3A4 (bicis.-3A4) microsomes as enzyme sources. This parallel approach significantly shortens sample preparation steps compared to an earlier manual processing with nanoparticle bioreactors, allowing a range of significant enzyme reactions to be processed simultaneously. Enzyme turnover rates using the microsomal bioreactors were 2-3 fold larger compared to using conventional microsomal dispersions, most likely because of better accessibility of the enzymes. Ketoconazole (KET) and quinidine (QIN), substrates specific to cyt P450 3A enzymes, were used to demonstrate applicability to establish potentially toxic drug-drug interactions involving enzyme inhibition and acceleration. PMID:19904994

  13. Ice Formation on Wings

    NASA Technical Reports Server (NTRS)

    Ritz, L

    1939-01-01

    This report makes use of the results obtained in the Gottingen ice tunnel in which the atmospheric conditions are simulated and the process of ice formation photographed. The effect of ice formation is threefold: 1) added weight to the airplane; 2) a change in the lift and drag forces; 3) a change in the stability characteristics.

  14. Formative Assessment Probes

    ERIC Educational Resources Information Center

    Eberle, Francis; Keeley, Page

    2008-01-01

    Formative assessment probes can be effective tools to help teachers build a bridge between students' initial ideas and scientific ones. In this article, the authors describe how using two formative assessment probes can help teachers determine the extent to which students make similar connections between developing a concept of matter and a…

  15. Formative Assessment in Context

    ERIC Educational Resources Information Center

    Oxenford-O'Brian, Julie

    2013-01-01

    This dissertation responds to critical gaps in current research on formative assessment practice which could limit successful implementation of this practice within the K-12 classroom context. The study applies a socio cultural perspective of learning to interpret a cross-case analysis of formative assessment practice occurring during one…

  16. School Formative Feedback Systems

    ERIC Educational Resources Information Center

    Halverson, Richard

    2010-01-01

    Data-driven instructional improvement relies on developing coherent systems that allow school staff to generate, interpret, and act upon quality formative information on students and school programs. This article offers a formative feedback system model that captures how school leaders and teachers structure artifacts and practices to create…

  17. Differential developmental toxicity of naphthoic acid isomers in medaka (Oryzias latipes) embryos.

    PubMed

    Carney, Michael W; Erwin, Kyle; Hardman, Ron; Yuen, Bonny; Volz, David C; Hinton, David E; Kullman, Seth W

    2008-01-01

    Polycyclic aromatic hydrocarbons (PAHs) are widespread persistent pollutants that readily undergo biotic and abiotic conversion to numerous transformation products in rivers, lakes and estuarine sediments. Here we characterize the developmental toxicity of four PAH transformation products each structural isomers of hydroxynaphthoic acid: 1H2NA, 2H1NA, 2H3NA, and 6H2NA. Medaka fish (Oryzias latipes) embryos and eleutheroembryos were used to determine toxicity. A 96-well micro-plate format was used to establish a robust, statistically significant platform for assessment of early life stages. Individual naphthoic acid isomers demonstrated a rank order of toxicity with 1H2NA>2H1NA>2H3NA>6H2NA being more toxic. Abnormalities of circulatory system were most pronounced including pericardial edema and tube heart. To determine if HNA isomers were AhR ligands, spatial-temporal expression and activity of CYP1A was measured via in vivo EROD assessments. qPCR measurement of CYP1A induction proved different between isomers dosed at respective concentrations affecting 50% of exposed individuals (EC50s). In vitro, all ANH isomers transactivated mouse AhR using a medaka CYP1A promoter specific reporter assay. Circulatory abnormalities followed P450 induction and response was consistent with PAH toxicity. A 96-well micro-plates proved suitable as exposure chambers and provided statistically sound evaluations as well as efficient toxicity screens. Our results demonstrate the use of medaka embryos for toxicity analysis thereby achieving REACH objectives for the reduction of adult animal testing in toxicity evaluations. PMID:18433798

  18. Fluorescent detection of Southern blots and PCR-based genetic typing tests

    SciTech Connect

    Mansfield, E.S.; Worley, J.M.; Zimmerman, P.A.

    1994-09-01

    The Southern blot is used to study gene organization, to identify disease-causing genomic rearrangements, or for typing RFLP markers in forensic, paternity, or prenatal diagnostic testing. Fluorescence offers a much greater dynamic range and a more linear response than film used in radioactive or chemiluminescent detection of RFLPs. We therefore investigated using the Fluorimager{trademark} 575 (Molecular Dynamics, Inc.) for analyzing Southern blots. Using a single-locus probe to D2S44 (YNH24) (Promega Corp.), we detect as little as 100 ng (0.05 attomole) genomic DNA. The alkaline phosphatase-labeled probe is detected using AttoPhos (JBL Scientific), and the developed membrane is scanned with the Fluorimager. Biotinylated hybridization probes can also be developed using a streptavidin-alkaline phosphatase conjugate and AttoPhos. The instrument scan parameters can be adjusted to prevent overexposure and accompanying loss of resolution in images of blots, gels, or 96-well microplates. We have used these other sample formats in PCR-based genetic typing assays. We use FluorKit DQS (Molecular Dynamics) to accurately quantify PCR template DNA (1-500 ng) in 96-well microplates scanned using the same instrument. Mutation detection assays run include heteroduplex gels (5% polyacrylamide, 2.7 M urea), short tandem repeat (STR) markers, amplified fragment length polymorphisms (AmpFLP), competitive priming PCR, and allele-specific oligotyping. These assays are run using either 1- or 2-color labeling. We detect unlabeled PCR products, such as the AmpFLP marker D1S80 (Perkin-Elmer) by post-staining gels for 10 minutes with SYBR Green 1 (Molecular Probes) and scanning the wet gel. The Fluorimager scans a 20 x 25 cm sample within three minutes, allowing rapid optimization of fluorescent protocols and high sample throughput.

  19. Inhibitory effects of the κ-casein macropeptide isolated from milk protein on the biofilm formation and virulence of Listeria monocytogenes.

    PubMed

    Yun, Hyun Sun; Kim, Younghoon; Park, Mi Ri; Kim, Sae Hun; Oh, Sejong

    2014-01-01

    We demonstrate the inhibitory effects of κ-casein macropeptide (CMP) on the biofilm formation and virulence of Listeria monocytogenes Scott A. The inhibition of biofilm formation by CMP was initially investigated by using the protocol applied for the 96-well microtiter plate assay. Low concentrations of CMP (0.1, 0.2, 0.3, 0.4, and 0.5 mg/mL) that were tested resulted in a profound inhibitory effect on biofilm formation at a concentration of 0.4 mg/mL. CMP also significantly repressed the transcription of inlA (encoding internalin A) that was responsible for the initial adhesion and invasion event, and prolonged the survival of Caenorhabditis elegans infected by L. monocytogenes. Two-dimensional gel electrophoresis showed that newly identified proteins in the presence of CMP were involved in the stress response and metabolic processes that have important roles in developing listerial biofilms. Our results suggest that CMP from milk protein would be capable of eliminating biofilm formation and virulence by L. monocytogenes in the food industry. PMID:25036840

  20. Inhibition of Serratia marcescens Smj-11 biofilm formation by Alcaligenes faecalis STN17 crude extract

    NASA Astrophysics Data System (ADS)

    Lutfi, Zainal; Usup, Gires; Ahmad, Asmat

    2014-09-01

    Serratia marcescens biofilms are formed when they are bound to surfaces in aqueous environments. S. marcescens utilizes N-acylhomoserine lactone (AHL) as its quorum sensing signal molecule. The accumulation of AHL indicates the bacteria to produce matrices to form biofilms. Prodigiosin (2-methyl-3-pentyl-6-methoxyprodigiosin), which causes red pigmentation in the colonies, are also produced when the AHL reaches a certain threshold. The Alcaligenes faecalis STN17 crude extract is believed to inhibit quorum sensing in the S. marcescens Smj-11 and, thus, impedes its biofilm formation ability. A. faecalis STN17 was grown in marine broth, and ethyl acetate extraction was carried out. The crude compound of A. faecalis STN17 was diluted at high concentration (0.2-6.4 mg/mL) and was taken to confirm anti-biofilm activity through the crystal violet method in 96-wells plate. Then, the crude extract underwent purification using simple solvents partitioning test to discern the respective compounds that had the anti-biofilm activity under the crystal violet method. The crystal violet test showed that the crude did have anti-biofilm activity on S. marcescens Smj-11, but did not kill the cells. This finding signifies that the suppression of biofilm formation in S. marcescens by A. faecalis STN17 has a strong correlation. The partitioning test showed that A. faecalis STN17 crude extract has several compounds and only the compound(s) in chloroform showed activities. In conclusion, the crude extract of A. faecalis STN17 has the ability to inhibit S. marcescens Smj-11 biofilm formation.

  1. Inhibition of Serratia marcescens Smj-11 biofilm formation by Alcaligenes faecalis STN17 crude extract

    SciTech Connect

    Lutfi, Zainal; Ahmad, Asmat; Usup, Gires

    2014-09-03

    Serratia marcescens biofilms are formed when they are bound to surfaces in aqueous environments. S. marcescens utilizes N-acylhomoserine lactone (AHL) as its quorum sensing signal molecule. The accumulation of AHL indicates the bacteria to produce matrices to form biofilms. Prodigiosin (2-methyl-3-pentyl-6-methoxyprodigiosin), which causes red pigmentation in the colonies, are also produced when the AHL reaches a certain threshold. The Alcaligenes faecalis STN17 crude extract is believed to inhibit quorum sensing in the S. marcescens Smj-11 and, thus, impedes its biofilm formation ability. A. faecalis STN17 was grown in marine broth, and ethyl acetate extraction was carried out. The crude compound of A. faecalis STN17 was diluted at high concentration (0.2-6.4 mg/mL) and was taken to confirm anti-biofilm activity through the crystal violet method in 96-wells plate. Then, the crude extract underwent purification using simple solvents partitioning test to discern the respective compounds that had the anti-biofilm activity under the crystal violet method. The crystal violet test showed that the crude did have anti-biofilm activity on S. marcescens Smj-11, but did not kill the cells. This finding signifies that the suppression of biofilm formation in S. marcescens by A. faecalis STN17 has a strong correlation. The partitioning test showed that A. faecalis STN17 crude extract has several compounds and only the compound(s) in chloroform showed activities. In conclusion, the crude extract of A. faecalis STN17 has the ability to inhibit S. marcescens Smj-11 biofilm formation.

  2. Sparse Image Format

    2007-04-12

    The Sparse Image Format (SIF) is a file format for storing spare raster images. It works by breaking an image down into tiles. Space is savid by only storing non-uniform tiles, i.e. tiles with at least two different pixel values. If a tile is completely uniform, its common pixel value is stored instead of the complete tile raster. The software is a library in the C language used for manipulating files in SIF format. Itmore » supports large files (> 2GB) and is designed to build in Windows and Linux environments.« less

  3. Sparse Image Format

    SciTech Connect

    Eads, Damian Ryan

    2007-04-12

    The Sparse Image Format (SIF) is a file format for storing spare raster images. It works by breaking an image down into tiles. Space is savid by only storing non-uniform tiles, i.e. tiles with at least two different pixel values. If a tile is completely uniform, its common pixel value is stored instead of the complete tile raster. The software is a library in the C language used for manipulating files in SIF format. It supports large files (> 2GB) and is designed to build in Windows and Linux environments.

  4. Biofilm formation and sanitizer resistance of Escherichia coli O157:H7 strains isolated from "high event period" meat contamination.

    PubMed

    Wang, Rong; Kalchayanand, Norasak; King, David A; Luedtke, Brandon E; Bosilevac, Joseph M; Arthur, Terrance M

    2014-11-01

    In the meat industry, a "high event period" (HEP) is defined as a time period during which commercial meat plants experience a higher than usual rate of Escherichia coli O157:H7 contamination. Genetic analysis indicated that within a HEP, most of the E. coli O157:H7 strains belong to a singular dominant strain type. This was in disagreement with the current beef contamination model stating that contamination occurs when incoming pathogen load on animal hides, which consists of diverse strain types of E. coli O157:H7, exceeds the intervention capacity. Thus, we hypothesize that the HEP contamination may be due to certain in-plant colonized E. coli O157:H7 strains that are better able to survive sanitization through biofilm formation. To test our hypothesis, a collection of 45 E. coli O157:H7 strains isolated from HEP beef contamination incidents and a panel of 47 E. coli O157:H7 strains of diverse genetic backgrounds were compared for biofilm formation and sanitizer resistance. Biofilm formation was tested on 96-well polystyrene plates for 1 to 6 days. Biofilm cell survival and recovery growth after sanitization were compared between the two strain collections using common sanitizers, including quaternary ammonium chloride, chlorine, and sodium chlorite. No difference in "early stage" biofilms was observed between the two strain collections after incubation at 22 to 25°C for 1 or 2 days. However, the HEP strains demonstrated significantly higher potency of "mature" biofilm formation after incubation for 4 to 6 days. Biofilms of the HEP strains also exhibited significantly stronger resistance to sanitization. These data suggest that biofilm formation and sanitization resistance could have a role in HEP beef contamination by E. coli O157:H7, which highlights the importance of proper and complete sanitization of food contact surfaces and food processing equipment in commercial meat plants. PMID:25364934

  5. Teaching Letter Formation.

    ERIC Educational Resources Information Center

    Graham, Steve; Madan, Avi J.

    1981-01-01

    The authors describe a remedial technique for teaching letter formation to students with handwriting difficulties. The approach blends traditional procedures (modeling, physical prompts, tracing, self correction, etc.) with cognitive behavior modification principles. (CL)

  6. Circumstellar grain formation

    NASA Technical Reports Server (NTRS)

    Draine, B. T.

    1986-01-01

    Dust formation around cool giant and supergiant stars is examined in terms of grain formulation. Optical properties of small clusters, molecular physics of cluster nucleation and growth, circumstellar mass flows, and their application to alpha Ori are discussed.

  7. Phenylpropanoids of plant origin as inhibitors of biofilm formation by Candida albicans.

    PubMed

    Raut, Jayant Shankar; Shinde, Ravikumar Bapurao; Chauhan, Nitin Mahendra; Karuppayil, Sankunny Mohan

    2014-09-01

    Biofilm-related infections of Candida albicans are a frequent cause of morbidity and mortality in hospitalized patients, especially those with immunocompromised status. Options of the antifungal drugs available for successful treatment of drug-resistant biofilms are very few, and as such, new strategies need to be explored against them. The aim of this study was to evaluate the efficacy of phenylpropanoids of plant origin against planktonic cells, important virulence factors, and biofilm forms of C. albicans. Standard susceptibility testing protocol was used to evaluate the activities of 13 phenylpropanoids against planktonic growth. Their effects on adhesion and yeast-to-hyphae morphogenesis were studied in microplate-based methodologies. An in vitro biofilm model analyzed the phenylpropanoid-mediated prevention of biofilm development and mature biofilms using XTT-metabolic assay, crystal violet assay, and light microscopy. Six molecules exhibited fungistatic activity at ≤0.5 mg/ml, of which four were fungicidal at low concentrations. Seven phenylpropanoids inhibited yeast-to-hyphae transition at low concentrations (0.031-0.5 mg/ml), whereas adhesion to the solid substrate was prevented in the range of 0.5-2 mg/ml. Treatment with ≤0.5 mg/ml concentrations of at least six small molecules resulted in significant (p < 0.05) inhibition of biofilm formation by C. albicans. Mature biofilms that are highly resistant to antifungal drugs were susceptible to low concentrations of 4 of the 13 molecules. This study revealed phenylpropanoids of plant origin as promising candidates to devise preventive strategies against drug-resistant biofilms of C. albicans. PMID:24851813

  8. Display formats manual

    NASA Technical Reports Server (NTRS)

    Runnels, R. L.

    1973-01-01

    The standards and procedures for the generation of operational display formats to be used in the Mission Control Center (MCC) display control system are presented. The required effort, forms, and fundamentals for the design, specifications, and production of display formats are identified. The principles of display design and system constraints controlling the creation of optimum operational displays for mission control are explained. The basic two types of MCC display systems for presenting information are described.

  9. Plant Formate Dehydrogenase

    SciTech Connect

    John Markwell

    2005-01-10

    The research in this study identified formate dehydrogenase, an enzyme that plays a metabolic role on the periphery of one-carbon metabolism, has an unusual localization in Arabidopsis thaliana and that the enzyme has an unusual kinetic plasticity. These properties make it possible that this enzyme could be engineered to attempt to engineer plants with an improved photosynthetic efficiency. We have produced transgenic Arabidopsis and tobacco plants with increased expression of the formate dehydrogenase enzyme to initiate further studies.

  10. Autonomous Formation Flight

    NASA Technical Reports Server (NTRS)

    Schkolnik, Gerard S.; Cobleigh, Brent

    2004-01-01

    NASA's Strategic Plan for the Aerospace Technology Enterprise includes ambitious objectives focused on affordable air travel, reduced emissions, and expanded aviation-system capacity. NASA Dryden Flight Research Center, in cooperation with NASA Ames Research Center, the Boeing Company, and the University of California, Los Angeles, has embarked on an autonomous-formation-flight project that promises to make significant strides towards these goals. For millions of years, birds have taken advantage of the aerodynamic benefit of flying in formation. The traditional "V" formation flown by many species of birds (including gulls, pelicans, and geese) enables each of the trailing birds to fly in the upwash flow field that exists just outboard of the bird immediately ahead in the formation. The result for each trailing bird is a decrease in induced drag and thus a reduction in the energy needed to maintain a given speed. Hence, for migratory birds, formation flight extends the range of the system of birds over the range of birds flying solo. The Autonomous Formation Flight (AFF) Project is seeking to extend this symbiotic relationship to aircraft.

  11. Multiplex profiling of cellular invasion in 3D cell culture models.

    PubMed

    Burgstaller, Gerald; Oehrle, Bettina; Koch, Ina; Lindner, Michael; Eickelberg, Oliver

    2013-01-01

    To-date, most invasion or migration assays use a modified Boyden chamber-like design to assess migration as single-cell or scratch assays on coated or uncoated planar plastic surfaces. Here, we describe a 96-well microplate-based, high-content, three-dimensional cell culture assay capable of assessing invasion dynamics and molecular signatures thereof. On applying our invasion assay, we were able to demonstrate significant effects on the invasion capacity of fibroblast cell lines, as well as primary lung fibroblasts. Administration of epidermal growth factor resulted in a substantial increase of cellular invasion, thus making this technique suitable for high-throughput pharmacological screening of novel compounds regulating invasive and migratory pathways of primary cells. Our assay also correlates cellular invasiveness to molecular events. Thus, we argue of having developed a powerful and versatile toolbox for an extensive profiling of invasive cells in a 96-well format. This will have a major impact on research in disease areas like fibrosis, metastatic cancers, or chronic inflammatory states. PMID:23671660

  12. Multiplex Profiling of Cellular Invasion in 3D Cell Culture Models

    PubMed Central

    Burgstaller, Gerald; Oehrle, Bettina; Koch, Ina; Lindner, Michael; Eickelberg, Oliver

    2013-01-01

    To-date, most invasion or migration assays use a modified Boyden chamber-like design to assess migration as single-cell or scratch assays on coated or uncoated planar plastic surfaces. Here, we describe a 96-well microplate-based, high-content, three-dimensional cell culture assay capable of assessing invasion dynamics and molecular signatures thereof. On applying our invasion assay, we were able to demonstrate significant effects on the invasion capacity of fibroblast cell lines, as well as primary lung fibroblasts. Administration of epidermal growth factor resulted in a substantial increase of cellular invasion, thus making this technique suitable for high-throughput pharmacological screening of novel compounds regulating invasive and migratory pathways of primary cells. Our assay also correlates cellular invasiveness to molecular events. Thus, we argue of having developed a powerful and versatile toolbox for an extensive profiling of invasive cells in a 96-well format. This will have a major impact on research in disease areas like fibrosis, metastatic cancers, or chronic inflammatory states. PMID:23671660

  13. A modified and automated version of the 'Fluorimetric Detection of Alkaline DNA Unwinding' method to quantify formation and repair of DNA strand breaks

    PubMed Central

    Moreno-Villanueva, María; Pfeiffer, Ragen; Sindlinger, Thilo; Leake, Alan; Müller, Marcus; Kirkwood, Thomas BL; Bürkle, Alexander

    2009-01-01

    Background Formation and repair of DNA single-strand breaks are important parameters in the assessment of DNA damage and repair occurring in live cells. The 'Fluorimetric Detection of Alkaline DNA Unwinding (FADU)' method [Birnboim HC, Jevcak JJ. Cancer Res (1981) 41:1889–1892] is a sensitive procedure to quantify DNA strand breaks, yet it is very tedious to perform. Results In order (i) to render the FADU assay more convenient and robust, (ii) to increase throughput, and (iii) to reduce the number of cells needed, we have established a modified assay version that is largely automated and is based on the use of a liquid handling device. The assay is operated in a 96-well format, thus greatly increasing throughput. The number of cells required has been reduced to less than 10,000 per data point. The threshold for detection of X-ray-induced DNA strand breaks is 0.13 Gy. The total assay time required for a typical experiment to assess DNA strand break repair is 4–5 hours. Conclusion We have established a robust and convenient method measuring of formation and repair of DNA single-strand breaks in live cells. While the sensitivity of our method is comparable to current assays, throughput is massively increased while operator time is decreased. PMID:19389244

  14. Positronium Formation in Muscle

    PubMed Central

    Gustafson, D. R.

    1970-01-01

    Positronium formation in muscle at +4°C and -4°C was examined by the measurement of the angular correlation of positron annihilation radiation. Since the positronium formation rate in ice is considerably higher than it is in water, there should be a comparable increase in the positronium formation rate in muscle tissue if recent speculation that cellular water is ordered in a semicrystalline icelike state is correct. Comparison of the angular correlation from muscle at +4°C with that from water at +4°C shows no enhancement of the positronium formation rate. Frozen muscle at -4°C shows an enhancement of the positronium formation rate of approximately half that found in ice at -4°C, indicating that most cellular water undergoes a normal water-ice transition when frozen. It is concluded therefore that cell water in muscle is not ordered in a hexagonal icelike structure. While the results are consistent with the hypothesis that cell water is in the liquid state, the hypothesis that cell water is ordered in an undetermined close packed structure which transforms to the hexagonal ice structure at or near 0°C cannot be ruled out. PMID:5436881

  15. Formation of the earth

    NASA Technical Reports Server (NTRS)

    Wetherill, George W.

    1990-01-01

    The origin of the earth is discussed in the context of the formation of the sun and the planets, and a standard model for such a formation assuming gravitational instability in a dense interstellar molecular cloud is outlined, along with the most significant variant of the model in which the loss of the nebular gas occurred after the formation of the earth. The formation of the sun and solar nebulae is addressed, and the coagulation of grains and the formation of small planetesimals are covered, along with the gravitational accumulation of planetesimals into planetary embryos and final stages of accumulation - embryos of planets. It is pointed out that the final stage of accumulation consists of the collision of these embryos; because of their large size, particularly after their further growth, these collisions represent giant impacts. It is concluded that the earth was initially an extremely hot and melted planet, surrounded by a fragile atmosphere and subject to violent impacts by bodies of the size of Ceres and even the moon.

  16. Molecules in star formation.

    NASA Astrophysics Data System (ADS)

    Shu, F. H.

    The author reviews current ideas and models in the problem of star formation from molecular cloud cores that are relatively isolated from the influences of other forming stars. He discusses the time scales, flow dynamics, and density and temperature structures applicable to each of the four stages of the entire process: (1) formation of a magnetized cloud core by ambipolar diffusion and evolution to a pivotal state of gravomagneto catastrophe; (2) self-similar collapse of the pivotal configuration and the formation of protostars, disks, and pseudo-disks; (3) onset of a magnetocentrifugally driven, lightly ionized wind from the interaction of an accretion disk and the magnetosphere of the central star, and the driving of bipolar molecular outflows; (4) evolution of pre-main-sequence stars surrounded by dusty accretion disks. For each of these stages and processes, he considers the characteristics of the molecular diagnostics needed to investigate the crucial aspects of the observational problem.

  17. Thrombus formation in vivo

    PubMed Central

    Furie, Bruce; Furie, Barbara C.

    2005-01-01

    To examine thrombus formation in a living mouse, new technologies involving intravital videomicroscopy have been applied to the analysis of vascular windows to directly visualize arterioles and venules. After vessel wall injury in the microcirculation, thrombus development can be imaged in real time. These systems have been used to explore the role of platelets, blood coagulation proteins, endothelium, and the vessel wall during thrombus formation. The study of biochemistry and cell biology in a living animal offers new understanding of physiology and pathology in complex biologic systems. PMID:16322780

  18. Crystal Formation in Inflammation.

    PubMed

    Franklin, Bernardo S; Mangan, Matthew S; Latz, Eicke

    2016-05-20

    The formation and accumulation of crystalline material in tissues is a hallmark of many metabolic and inflammatory conditions. The discovery that the phase transition of physiologically soluble substances to their crystalline forms can be detected by the immune system and activate innate immune pathways has revolutionized our understanding of how crystals cause inflammation. It is now appreciated that crystals are part of the pathogenesis of numerous diseases, including gout, silicosis, asbestosis, and atherosclerosis. In this review we discuss current knowledge of the complex mechanisms of crystal formation in diseased tissues and their interplay with the nutrients, metabolites, and immune cells that account for crystal-induced inflammation. PMID:26772211

  19. Gaussian entanglement of formation

    SciTech Connect

    Wolf, M.M.; Giedke, G.; Krueger, O.; Werner, R. F.; Cirac, J.I.

    2004-05-01

    We introduce a Gaussian version of the entanglement of formation adapted to bipartite Gaussian states by considering decompositions into pure Gaussian states only. We show that this quantity is an entanglement monotone under Gaussian operations and provide a simplified computation for states of arbitrary many modes. For the case of one mode per site the remaining variational problem can be solved analytically. If the considered state is in addition symmetric with respect to interchanging the two modes, we prove additivity of the considered entanglement measure. Moreover, in this case and considering only a single copy, our entanglement measure coincides with the true entanglement of formation.

  20. Isolating Triggered Star Formation

    SciTech Connect

    Barton, Elizabeth J.; Arnold, Jacob A.; Zentner, Andrew R.; Bullock, James S.; Wechsler, Risa H.; /KIPAC, Menlo Park /SLAC

    2007-09-12

    Galaxy pairs provide a potentially powerful means of studying triggered star formation from galaxy interactions. We use a large cosmological N-body simulation coupled with a well-tested semi-analytic substructure model to demonstrate that the majority of galaxies in close pairs reside within cluster or group-size halos and therefore represent a biased population, poorly suited for direct comparison to 'field' galaxies. Thus, the frequent observation that some types of galaxies in pairs have redder colors than 'field' galaxies is primarily a selection effect. We use our simulations to devise a means to select galaxy pairs that are isolated in their dark matter halos with respect to other massive subhalos (N= 2 halos) and to select a control sample of isolated galaxies (N= 1 halos) for comparison. We then apply these selection criteria to a volume-limited subset of the 2dF Galaxy Redshift Survey with M{sub B,j} {le} -19 and obtain the first clean measure of the typical fraction of galaxies affected by triggered star formation and the average elevation in the star formation rate. We find that 24% (30.5 %) of these L* and sub-L* galaxies in isolated 50 (30) h{sup -1} kpc pairs exhibit star formation that is boosted by a factor of {approx}> 5 above their average past value, while only 10% of isolated galaxies in the control sample show this level of enhancement. Thus, 14% (20 %) of the galaxies in these close pairs show clear triggered star formation. Our orbit models suggest that 12% (16%) of 50 (30) h{sup -1} kpc close pairs that are isolated according to our definition have had a close ({le} 30 h{sup -1} kpc) pass within the last Gyr. Thus, the data are broadly consistent with a scenario in which most or all close passes of isolated pairs result in triggered star formation. The isolation criteria we develop provide a means to constrain star formation and feedback prescriptions in hydrodynamic simulations and a very general method of understanding the importance of

  1. BISAC Variable Format.

    ERIC Educational Resources Information Center

    Information Technology and Libraries, 1983

    1983-01-01

    Presents revision of Book Industry Systems Advisory Committee (BISAC) format designed specifically for electronic transmission of purchase orders for monograph or series titles combining fixed and variable length data fields which was approved in January 1983. Special characters, sample address descriptions, summary of fixed records, glossary, and…

  2. The Formation of Trihalomethanes.

    ERIC Educational Resources Information Center

    Trussell, R. Rhodes; Umphres, Mark D.

    1978-01-01

    Reviewed are a number of factors important in the formation of trihalomethanes (THM) including the nature of aquatic humus and the influences of preozonation, bromide, pH, and chlorine. A brief investigation is also conducted into the kinetics of the THM reaction. Several major research needs are represented. (CS)

  3. Reconsidering Formative Measurement

    ERIC Educational Resources Information Center

    Howell, Roy D.; Breivik, Einar; Wilcox, James B.

    2007-01-01

    The relationship between observable responses and the latent constructs they are purported to measure has received considerable attention recently, with particular focus on what has become known as formative measurement. This alternative to reflective measurement in the area of theory-testing research is examined in the context of the potential…

  4. Formation of Freirian Facilitators.

    ERIC Educational Resources Information Center

    Noble, Phyllis

    This paper is written for people who are already familiar with the philosophy and methodology of Paulo Freire's liberatory education and are interested in creating a formation program for adult education facilitators using his ideas. The author describes the paper as "a collection of thoughts, of things to consider," when organizing such a…

  5. Formation of planetesimals

    NASA Technical Reports Server (NTRS)

    Weidenschilling, Stuart J.

    1991-01-01

    Formation of planetesimals is discussed. The following subject areas are covered: (1) nebular structure; (2) aerodynamics of the solid bodies in the nebula; (3) problems with gravitational instability; (4) particle growth by coagulation; properties of fractal aggregates; and (5) coagulation and settling of fractal aggregates.

  6. Technobabble: File Formats.

    ERIC Educational Resources Information Center

    Wilson, Bradley

    1999-01-01

    Considers the confusion of over 20 different kinds of graphics programs. Briefly distinguishes between some of the more popular graphics formats (Photoshop, TIFF, JPEG, GIF, PICT, and EPS), and describes the benefits and disadvantages of each in the context of journalism education. (SC)

  7. FORMATION OF PHOTOCHEMICAL AEROSOLS

    EPA Science Inventory

    The objective was to develop a better understanding of smog aerosol formation with particular reference to haze in the Southern California area. This study combined laboratory work with ambient air studies. Counting of particles by light scattering was the principle physical tech...

  8. Common file formats.

    PubMed

    Leonard, Shonda A; Littlejohn, Timothy G; Baxevanis, Andreas D

    2007-01-01

    This appendix discusses a few of the file formats frequently encountered in bioinformatics. Specifically, it reviews the rules for generating FASTA files and provides guidance for interpreting NCBI descriptor lines, commonly found in FASTA files. In addition, it reviews the construction of GenBank, Phylip, MSF and Nexus files. PMID:18428774

  9. Bacterial formate hydrogenlyase complex

    PubMed Central

    McDowall, Jennifer S.; Murphy, Bonnie J.; Haumann, Michael; Palmer, Tracy; Armstrong, Fraser A.; Sargent, Frank

    2014-01-01

    Under anaerobic conditions, Escherichia coli can carry out a mixed-acid fermentation that ultimately produces molecular hydrogen. The enzyme directly responsible for hydrogen production is the membrane-bound formate hydrogenlyase (FHL) complex, which links formate oxidation to proton reduction and has evolutionary links to Complex I, the NADH:quinone oxidoreductase. Although the genetics, maturation, and some biochemistry of FHL are understood, the protein complex has never been isolated in an intact form to allow biochemical analysis. In this work, genetic tools are reported that allow the facile isolation of FHL in a single chromatographic step. The core complex is shown to comprise HycE (a [NiFe] hydrogenase component termed Hyd-3), FdhF (the molybdenum-dependent formate dehydrogenase-H), and three iron-sulfur proteins: HycB, HycF, and HycG. A proportion of this core complex remains associated with HycC and HycD, which are polytopic integral membrane proteins believed to anchor the core complex to the cytoplasmic side of the membrane. As isolated, the FHL complex retains formate hydrogenlyase activity in vitro. Protein film electrochemistry experiments on Hyd-3 demonstrate that it has a unique ability among [NiFe] hydrogenases to catalyze production of H2 even at high partial pressures of H2. Understanding and harnessing the activity of the FHL complex is critical to advancing future biohydrogen research efforts. PMID:25157147

  10. Kepler Planet Formation

    NASA Technical Reports Server (NTRS)

    Lissauer, Jack J.

    2015-01-01

    Kepler has vastly increased our knowledge of planets and planetary systems located close to stars. The new data shows surprising results for planetary abundances, planetary spacings and the distribution of planets on a mass-radius diagram. The implications of these results for theories of planet formation will be discussed.

  11. 500+ Writing Formats.

    ERIC Educational Resources Information Center

    McIntosh, Margaret E.

    1997-01-01

    Suggests a multitude of ideas for students to communicate their ideas in writing using the language of mathematics. Includes a sampling of 500+ writing formats, 67 abbreviated writing assignments, and three complete assignments along with a sample student response to each. Sample assignments include advice column, biographical sketch, commercial,…

  12. Formation in the Classroom

    ERIC Educational Resources Information Center

    Glennon, Fred; Jacobsen, Douglas; Jacobsen, Rhonda Hustedt; Thatamanil, John J.; Porterfield, Amanda; Moore, Mary Elizabeth

    2011-01-01

    What is the relationship between the academic knowledge of the guild and the formation of students in the classroom? This Forum gathers four essays originally presented at a Special Topics Session at the 2009 conference of the American Academy of Religion (Atlanta, Georgia), with a brief introductory essay by Fred Glennon explaining the genesis of…

  13. Insights on galaxy formation

    NASA Astrophysics Data System (ADS)

    Bullock, James Steven

    1999-12-01

    Recent advances in theoretical modeling coupled with a wealth of new observational data, provide a unique opportunity for gaining insight into process of galaxy formation. I present results which test and develop current theories. The analysis utilizes state of the art theoretical modeling and makes predictions aimed at comparisons with some of the latest and upcoming observational data sets. In part I, I discuss an analysis of the structure and properties of dark matter halos (believed to govern the dynamical evolution of galaxies). The results make use of very high-resolution N-body simulations, and are derived from a new hierarchical halo finder, designed especially for these projects and to complement advancements in simulation technology. I present information on the dark matter halo substructure, density profiles, angular momentum structure, and collision rates. In part II, I discuss some aspects of galaxy formation theory in light of new observational data. The discussion includes an investigation of the nature of high-redshift galaxies, the local velocity function of galaxies, and the use of gamma ray telescopes to probe the extra-galactic background light-the latter analysis is done in the context of semi-analytic modeling of galaxy formation. The most important conclusions of this thesis are as follows. (1)Dark matter halos at high redshift are much less concentrated than previously believed. implying that quiescently star-forming galaxies at high redshift are larger and dimmer than expected. (2)The observed bright. abundant. and highly clustered high- redshift (Lyman-break) galaxies are likely starbursts driven by collisions between relatively small galaxies at z ~ 3. And (3)there is a real possibility of using the growing advances in γ-ray astronomy to probe many poorly constrained processes of galaxy formation, including the stellar initial mass function and the star formation history of the universe.

  14. Pattern formation today

    PubMed Central

    Chuong, Cheng-Ming; Richardson, Michael K.

    2010-01-01

    Patterns are orders embedded in randomness. They may appear as spatial arrangements or temporal series, and the elements may appear identical or with variations. Patterns exist in the physical world as well as in living systems. In the biological world, patterns can range from simple to complex, forming the basic building blocks of life. The process which generates this ordering in the biological world was termed pattern formation. Since Wolpert promoted this concept four decades ago, scientists from molecular biology, developmental biology, stem cell biology, tissue engineering, theoretical modeling and other disciplines have made remarkable progress towards understanding its mechanisms. It is time to review and re-integrate our understanding. Here, we explore the origin of pattern formation, how the genetic code is translated into biological form, and how complex phenotypes are selected over evolutionary time. We present four topics: Principles, Evolution, Development, and Stem Cells and Regeneration. We have interviewed several leaders in the field to gain insight into how their research and the field of pattern formation have shaped each other. We have learned that both molecular process and physico-chemical principles are important for biological pattern formation. New understanding will emerge through integration of the analytical approach of molecular-genetic manipulation and the systemic approach of model simulation. We regret that we could not include every major investigator in the field, but hope that this Special Issue of the Int. J. Dev. Biol. represents a sample of our knowledge of pattern formation today, which will help to stimulate more research on this fundamental process. PMID:19557673

  15. Medusae Fossae Formation

    NASA Technical Reports Server (NTRS)

    2002-01-01

    (Released 10 April 2002) The Science This THEMIS visible image was acquired near 7o S, 172o W (188o E) and shows a remarkable martian geologic deposit known as the Medusae Fossae Formation. This Formation, seen here as the raised plateau in the upper two-thirds of the image, is a soft, easily eroded deposit that extends for nearly 1,000 km along the equator of Mars. In this region the deposit has been heavily eroded by the wind to produce a series of linear ridges called yardangs. These parallel ridges point in direction of the prevailing winds that carved them, and demonstrate the power of martian winds to sculpt the dry landscape of Mars. The Medusae Fossae Formation has been completely stripped from the surface in the lower third of the image, revealing a harder layer below that is more resistant to wind erosion. The easily eroded nature of the Medusae Fossae Formation suggests that it is composed of weakly cemented particles, and was most likely formed by the deposition of wind-blown dust or volcanic ash. Several ancient craters that were once completely buried by this deposit are being exposed, or exhumed, as the overlying Medusae Formation is removed. Very few impact craters are visible on this Formation, indicating that the surface seen today is relatively young, and that the processes of erosion are likely to be actively occurring. The Story Medusa of Greek mythology fame, the name-giver to this region, had snaky locks of hair that could turn a person to stone. Wild and unruly, this monster of the underworld could certainly wreak havoc on the world of the human imagination. As scary as she was, Medusa would have no advantage over the fierce, masterful winds blowing across Mars, which once carved the streaky, terrain at the top of this image. Wild and whipping, these winds have slowly eroded away the 'topsoil,' revealing ancient craters and other surface features they once covered. The loosely cemented particles of this 'topsoil' are likely made up of dust

  16. Format( )MEDIC( )Input

    NASA Astrophysics Data System (ADS)

    Foster, K.

    1994-09-01

    This document is a description of a computer program called Format( )MEDIC( )Input. The purpose of this program is to allow the user to quickly reformat wind velocity data in the Model Evaluation Database (MEDb) into a reasonable 'first cut' set of MEDIC input files (MEDIC.nml, StnLoc.Met, and Observ.Met). The user is cautioned that these resulting input files must be reviewed for correctness and completeness. This program will not format MEDb data into a Problem Station Library or Problem Metdata File. A description of how the program reformats the data is provided, along with a description of the required and optional user input and a description of the resulting output files. A description of the MEDb is not provided here but can be found in the RAS Division Model Evaluation Database Description document.

  17. Pattern formation during vasculogenesis.

    PubMed

    Czirok, Andras; Little, Charles D

    2012-06-01

    Vasculogenesis, the assembly of the first vascular network, is an intriguing developmental process that yields the first functional organ system of the embryo. In addition to being a fundamental part of embryonic development, vasculogenic processes also have medical importance. To explain the organizational principles behind vascular patterning, we must understand how morphogenesis of tissue level structures can be controlled through cell behavior patterns that, in turn, are determined by biochemical signal transduction processes. Mathematical analyses and computer simulations can help conceptualize how to bridge organizational levels and thus help in evaluating hypotheses regarding the formation of vascular networks. Here, we discuss the ideas that have been proposed to explain the formation of the first vascular pattern: cell motility guided by extracellular matrix alignment (contact guidance), chemotaxis guided by paracrine and autocrine morphogens, and sprouting guided by cell-cell contacts. PMID:22692888

  18. Cosmological structure formation

    NASA Technical Reports Server (NTRS)

    Schramm, David N.

    1991-01-01

    A summary of the current forefront problem of physical cosmology, the formation of structures (galaxies, clusters, great walls, etc.) in the universe is presented. Solutions require two key ingredients: (1) matter; and (2) seeds. Regarding the matter, it now seems clear that both baryonic and non-baryonic matter are required. Whether the non-baryonic matter is hot or cold depends on the choice of seeds. Regarding the seeds, both density fluctuations and topological defects are discussed. The combination of isotropy of the microwave background and the recent observations indicating more power on large scales have severly constrained, if not eliminated, Gaussian fluctuations with equal power on all scales, regardless of the eventual resolution of both the matter and seed questions. It is important to note that all current structure formation ideas require new physics beyond SU(3) x SU(2) x U(1).

  19. Emptiness Formation Probability

    NASA Astrophysics Data System (ADS)

    Crawford, Nicholas; Ng, Stephen; Starr, Shannon

    2016-08-01

    We present rigorous upper and lower bounds on the emptiness formation probability for the ground state of a spin-1/2 Heisenberg XXZ quantum spin system. For a d-dimensional system we find a rate of decay of the order {exp(-c L^{d+1})} where L is the sidelength of the box in which we ask for the emptiness formation event to occur. In the {d=1} case this confirms previous predictions made in the integrable systems community, though our bounds do not achieve the precision predicted by Bethe ansatz calculations. On the other hand, our bounds in the case {d ≥ 2} are new. The main tools we use are reflection positivity and a rigorous path integral expansion, which is a variation on those previously introduced by Toth, Aizenman-Nachtergaele and Ueltschi.

  20. Formation of bacterial nanocells

    NASA Astrophysics Data System (ADS)

    Vainshtein, Mikhail; Kudryashova, Ekaterina; Suzina, Natalia; Ariskina, Elena; Voronkov, Vadim

    1998-07-01

    Existence of nanobacteria received increasing attention both in environmental microbiology/geomicro-biology and in medical microbiology. In order to study a production of nanoforms by typical bacterial cells. Effects of different physical factors were investigated. Treatment of bacterial cultures with microwave radiation, or culturing in field of electric current resulted in formation a few types of nanocells. The number and type of nanoforms were determined with type and dose of the treatment. The produced nanoforms were: i) globules, ii) clusters of the globules--probably produced by liaison, iii) nanocells coated with membrane. The viability of the globules is an object opened for doubts. The nanocells discovered multiplication and growth on solidified nutrient media. The authors suggest that formation of nanocells is a common response of bacteria to stress-actions produced by different agents.

  1. Prominence Formation Processes

    NASA Astrophysics Data System (ADS)

    Welsch, B. T.; DeVore, C. R.; Antiochos, S. K.

    2005-01-01

    Martens and Zwaan (ApJ v. 558 872) have proposed a prominence/ filament formation model in which differential rotation drives reconnection between two initially unconnected active regions to form helical field lines that support mass and are held down by overlying field. Using an MHD solver with adaptive refinement we simulated this process by imposing a shear flow meant to mimic differential rotation on two bipolar flux distributions meant to mimic distinct active regions. In some runs the flux systems are initially potential while in others they have been twisted by footpoint rotation to inject helicity prior to imposing the shear flow. The resulting structures are studied to understand the role of helicity in the formation of prominence-like structures.

  2. Format-Preserving Encryption

    NASA Astrophysics Data System (ADS)

    Bellare, Mihir; Ristenpart, Thomas; Rogaway, Phillip; Stegers, Till

    Format-preserving encryption (FPE) encrypts a plaintext of some specified format into a ciphertext of identical format—for example, encrypting a valid credit-card number into a valid credit-card number. The problem has been known for some time, but it has lacked a fully general and rigorous treatment. We provide one, starting off by formally defining FPE and security goals for it. We investigate the natural approach for achieving FPE on complex domains, the “rank-then-encipher” approach, and explore what it can and cannot do. We describe two flavors of unbalanced Feistel networks that can be used for achieving FPE, and we prove new security results for each. We revisit the cycle-walking approach for enciphering on a non-sparse subset of an encipherable domain, showing that the timing information that may be divulged by cycle walking is not a damaging thing to leak.

  3. Formate-assisted pyrolysis

    SciTech Connect

    DeSisto, William Joseph; Wheeler, Marshall Clayton; van Heiningen, Adriaan R. P.

    2015-03-17

    The present invention provides, among other thing, methods for creating significantly deoxygenated bio-oils form biomass including the steps of providing a feedstock, associating the feedstock with an alkali formate to form a treated feedstock, dewatering the treated feedstock, heating the dewatered treated feedstock to form a vapor product, and condensing the vapor product to form a pyrolysis oil, wherein the pyrolysis oil contains less than 30% oxygen by weight.

  4. Hail Formation in Florida

    NASA Astrophysics Data System (ADS)

    Stanley, Matthew

    Hail poses a substantial threat to life and property in the state of Florida. These losses could be minimized through better understanding of the relationships between atmospheric variables that impact hail formation in Florida. Improving hail forecasting in Florida requires analyzing a number of meteorological parameters and synoptic data related to hail formation. NOAA archive data was retrieved to create a database that was used to categorize text files of hail days. The text files were entered into the National Oceanic and Atmospheric Administration Earth System Research Laboratory website to create National Centers for Environmental Prediction/National Center for Atmospheric Research Reanalysis maps of atmospheric variables for Florida hail days as well as days leading to the hail event. These data were then analyzed to determine the relationship between variables that affect hail formation, in general, across different regions and seasons in Florida using Statistical Product and Service Solutions. The reasoning for the differing factors affecting hail formation between regions, seasons and hail sizes were discussed, as well as forecasting suggestions relating to region and month in Florida. The study found that the majority of all hail that occurs in Florida is during the wet season. A low Lifted Index, high Precipitable Water and lower than average Sea Level Pressure, in most cases, is present during hail days in Florida. Furthermore, results show that Vector Wind magnitude increases as hail size increases. Additionally, several atmospheric variables useful to studying hail events, such as Lifted Index, Precipitable Water, Sea Level Pressure, Vector Wind and Temperature have significant correlations with each other depending on the region and season being observed. Strong correlations between low Lifted Index, high Precipitable Water values and the occurrence of hail events are discussed, as well as the relationship between temperature anomalies at various

  5. Galaxy Formation and Evolution

    NASA Astrophysics Data System (ADS)

    Nagamine, Kentaro; Reddy, Naveen; Daddi, Emanuele; Sargent, Mark T.

    2016-07-01

    In this chapter, we discuss the current status of observational and computational studies on galaxy formation and evolution. In particular, a joint analysis of star-formation rates (SFRs), stellar masses, and metallicities of galaxies throughout cosmic time can shed light on the processes by which galaxies build up their stellar mass and enrich the environment with heavy elements. Comparison of such observations and the results of numerical simulations can give us insights on the physical importance of various feedback effects by supernovae and active galactic nuclei. In Sect. 1, we first discuss the primary methods used to deduce the SFRs, stellar masses, and (primarily) gas-phase metallicities in high-redshift galaxies. Then, we show how these quantities are related to each other and evolve with time. In Sect. 2, we further examine the distribution of SFRs in galaxies following the `Main Sequence' paradigm. We show how the so-called `starbursts' display higher specific SFRs and SF efficiencies by an order of magnitude. We use this to devise a simple description of the evolution of the star-forming galaxy population since z ˜3 that can successfully reproduce some of the observed statistics in the infrared (IR) wavelength. We also discuss the properties of molecular gas. In Sect. 3, we highlight some of the recent studies of high-redshift galaxy formation using cosmological hydrodynamic simulations. We discuss the physical properties of simulated galaxies such as luminosity function and escape fraction of ionizing photons, which are important statistics for reionization of the Universe. In particular the escape fraction of ionizing photons has large uncertainties, and studying gamma-ray bursts (which is the main topic of this conference) can also set observational constraints on this uncertain physical parameter as well as cosmic star formation rate density.

  6. Mesospheric cloud formations

    NASA Technical Reports Server (NTRS)

    Forbes, J. M.

    1980-01-01

    Formation of mesospheric clouds as a result of deposition of large amounts of H2O by the heavy lift launch vehicle (HLLV) of the solar power satellite system is discussed. The conditions which must be met in order to form and maintain clouds near the mesopause are described. The frequency and magnitude of H2O injections from the HLLV rocket exhaust are considered.

  7. Formation of Bulges

    NASA Astrophysics Data System (ADS)

    Silk, Joseph; Bouwens, Rychard

    1999-07-01

    Bulges, often identified with the spheroidal component of a galaxy,have a complex pedigree. Massive bulges are generally red and old,but lower mass bulges have broader dispersions in color that may becorrelated with disk colors. This suggests different formationscenarios. I will review possible formation sequences for bulges,describe the various signatures that distinguish these scenarios, anddiscuss implications for the high redshift universe.

  8. Terrestrial planet formation

    PubMed Central

    Righter, K.; O’Brien, D. P.

    2011-01-01

    Advances in our understanding of terrestrial planet formation have come from a multidisciplinary approach. Studies of the ages and compositions of primitive meteorites with compositions similar to the Sun have helped to constrain the nature of the building blocks of planets. This information helps to guide numerical models for the three stages of planet formation from dust to planetesimals (∼106 y), followed by planetesimals to embryos (lunar to Mars-sized objects; few × 106 y), and finally embryos to planets (107–108 y). Defining the role of turbulence in the early nebula is a key to understanding the growth of solids larger than meter size. The initiation of runaway growth of embryos from planetesimals ultimately leads to the growth of large terrestrial planets via large impacts. Dynamical models can produce inner Solar System configurations that closely resemble our Solar System, especially when the orbital effects of large planets (Jupiter and Saturn) and damping mechanisms, such as gas drag, are included. Experimental studies of terrestrial planet interiors provide additional constraints on the conditions of differentiation and, therefore, origin. A more complete understanding of terrestrial planet formation might be possible via a combination of chemical and physical modeling, as well as obtaining samples and new geophysical data from other planets (Venus, Mars, or Mercury) and asteroids. PMID:21709256

  9. Tetrahedron Formation Control

    NASA Technical Reports Server (NTRS)

    Guzman, Jose J.

    2003-01-01

    Spacecraft flying in tetrahedron formations are excellent instrument platforms for electromagnetic and plasma studies. A minimum of four spacecraft - to establish a volume - is required to study some of the key regions of a planetary magnetic field. The usefulness of the measurements recorded is strongly affected by the tetrahedron orbital evolution. This paper considers the preliminary development of a general optimization procedure for tetrahedron formation control. The maneuvers are assumed to be impulsive and a multi-stage optimization method is employed. The stages include targeting to a fixed tetrahedron orientation, rotating and translating the tetrahedron and/or varying the initial and final times. The number of impulsive maneuvers citn also be varied. As the impulse locations and times change, new arcs are computed using a differential corrections scheme that varies the impulse magnitudes and directions. The result is a continuous trajectory with velocity discontinuities. The velocity discontinuities are then used to formulate the cost function. Direct optimization techniques are employed. The procedure is applied to the Magnetospheric Multiscale Mission (MMS) to compute preliminary formation control fuel requirements.

  10. Mars brine formation experiment

    NASA Technical Reports Server (NTRS)

    Moore, Jeffrey M.; Bullock, Mark A.; Stoker, Carol R.

    1992-01-01

    Evaporites, particularly carbonates, nitrates, and sulfates, may be major sinks of volatiles scavenged from the martian atmosphere. Mars is thought to have once had a denser, warmer atmosphere that permitted the presence of liquid surface water. The conversion of atmospheric CO2 into carbonate is hypothesized to have degraded the martian climate to its present state of a generally subfreezing, desiccated desert. The rate for such a conversion under martian conditions is poorly known, so the time scale of climate degradation by this process cannot be easily evaluated. If some models are correct, carbonate formation may have been fast at geological time scales. The experiments of Booth and Kieffer also imply fast (10(exp 6) - 10(exp 7) yr) removal of the missing CO2 inventory, estimated to be 1 - 5 bar, by means of carbonate formation. The timing of formation of many of the fluvial features observed on Mars is, in large part, dependent on when and how fast the atmosphere changed. A knowledge of the rate at which carbonates and nitrates formed is also essential for assessing the probability that life, or its chemical precursors, could have developed on Mars. No previous experiments have quantitatively evaluated the rate of solution for a suite of mobile anions and cations from unaltered minerals and atmospheric gases into liquid water under Mars-like conditions. Such experiments are the focus of this task.

  11. Terrestrial planet formation.

    PubMed

    Righter, K; O'Brien, D P

    2011-11-29

    Advances in our understanding of terrestrial planet formation have come from a multidisciplinary approach. Studies of the ages and compositions of primitive meteorites with compositions similar to the Sun have helped to constrain the nature of the building blocks of planets. This information helps to guide numerical models for the three stages of planet formation from dust to planetesimals (~10(6) y), followed by planetesimals to embryos (lunar to Mars-sized objects; few 10(6) y), and finally embryos to planets (10(7)-10(8) y). Defining the role of turbulence in the early nebula is a key to understanding the growth of solids larger than meter size. The initiation of runaway growth of embryos from planetesimals ultimately leads to the growth of large terrestrial planets via large impacts. Dynamical models can produce inner Solar System configurations that closely resemble our Solar System, especially when the orbital effects of large planets (Jupiter and Saturn) and damping mechanisms, such as gas drag, are included. Experimental studies of terrestrial planet interiors provide additional constraints on the conditions of differentiation and, therefore, origin. A more complete understanding of terrestrial planet formation might be possible via a combination of chemical and physical modeling, as well as obtaining samples and new geophysical data from other planets (Venus, Mars, or Mercury) and asteroids. PMID:21709256

  12. Model of kimberlite formation

    NASA Astrophysics Data System (ADS)

    Kostrovitsky, Sergey; Fiveyskaya, Lyudmila

    2013-04-01

    The critical goals in recognizing the nature of kimberlites are to find out: (1) the primary composition of melt of these rocks and (2) the principal processes of evolution of primary composition of kimberlites while ascending from mantle depth towards earth surface. Suppose, that the primary composition of kimberlite melt-fluid was in fact the composition of asthenosphere melt geochemically being close to alkaline-basalt (Hi-µ) saturated with high CO2. The genetic relation of kimberlites with basaltoids is indicated by a spatial and temporal affinity of their formation (Carlson et al, 2006; Lehmann et al, 2010; Tappe et al, 2012), similarity of the pattern of incompatible elements distribution, presence of megacryst minerals in alkaline basaltoids, Pyr-Alm garnet included, and finally, model calculation of parent melt composition for low-Cr megacryst minerals; it showed this composition to be typical for the alkaline basaltoid (Jones, 1980). At the asthenosphere level there was differentiation of basaltoid melt-fluid which was responsible for formation of its different parts with varying melt to fluid ratio and possibly varying content of alkalis (K2O). The outbreak of asthenosphere substance through lithosphere mantle proceeded by different scenarios: (a) With a noticeable dominance of fluid component kimberlites were formed by the capture and contamination of high-Mg, high-Cr rocks of lithosphere mantle that caused formation of high-Mg kimberlites. That corresponds to model of Russell (2012). (b) With a considerable proportion of melt phase depending on saturation in fluid there formed magnesium-ferriferous and ferriferous-titaniferous petrochemical types of kimberlites. There is no doubt that in formation of these kimberlite types the contamination of lithosphere material was the case, at the much lower level than in formation of high-Mg kimberlites. This model logically explains steady differences of petrochemistry of kimberlites making up clusters of

  13. Formation of giant planets

    NASA Astrophysics Data System (ADS)

    Magni, G.; Coradini, A.

    2003-04-01

    In this presentation we address the problem of the formation of giant planets and their regular satellites. We study in particular the problem of formation of the Jupiter System comparing the results of the model with the present characteristics of the system, in order to identify what are those better represented by our approach. In fact here, using a 3-D hydro-dynamical code, we study the modalities of gas accretion onto a solid core, believed to be the seed from which Jupiter started. To do that we have modelled three main regions: the central planet, a turbulent accretion disk surrounding it and an extended region from which the gas is collected. In the extended region we treat the gas as a frictionless fluid. Our main goal is to identify what are the characteristics of the planet during its growth and the physical parameters affecting its growth at the expenses of the nebular gas present in the feeding zone. Moreover we want to understand what are the thermodynamical parameters characterizing the gas captured by the planet and swirling around it. Finally, we check if a disk can be formed in prograde rotation around the planet and if this disk can survive the final phases of the planet formation. Due to the interaction between the accreting planet and the disk it has been necessary to develop a complete model of the Jupiter’s structure. In fact the radiation emitted by the growing planet heats up the surrounding gas. In turn the planet’s thermodynamic structure depend on the mass accretion rate onto it. When the accretion is rapid, shock waves in the gas are formed close to the planet. This region cannot be safely treated by a numerical code; for this reason we have developed a semi-analytically model of a a turbulent accretion disk to be considered as transition between the planet and the surrounding disk.

  14. Liposome formation in microgravity

    NASA Astrophysics Data System (ADS)

    Claassen, D. E.; Spooner, B. S.

    Liposomes are artificial vesicles with a phospholipid bilayer membrane. The formation of liposomes is a self-assembly process that is driven by the amphipathic nature of phospholipid molecules and can be observed during the removal of detergent from phospholipids dissolved in detergent micelles. As detergent concentration in the mixed micelles decreases, the non-polar tail regions of phospholipids produce a hydrophobic effect that drives the micelles to fuse and form planar bilayers in which phospholipids orient with tail regions to the center of the bilayer and polar head regions to the external surface. Remaining detergent molecules shield exposed edges of the bilayer sheet from the aqueous environment. Further removal of detergent leads to intramembrane folding and membrane vesiculation, forming liposomes. We have observed that the formation of liposomes is altered in microgravity. Liposomes that were formed at 1-g did not exceed 150 nm in diameter, whereas liposomes that were formed during spaceflight exhibited diameters up to 2000 nm. Using detergent-stabilized planar bilayers, we determined that the stage of liposome formation most influenced by gravity is membrane vesiculation. In addition, we found that small, equipment-induced fluid disturbances increased vesiculation and negated the size-enhancing effects of microgravity. However, these small disturbances had no effect on liposome size at 1-g, likely due to the presence of gravity-induced buoyancy-driven fluid flows (e.g., convection currents). Our results indicate that fluid disturbances, induced by gravity, influence the vesiculation of membranes and limit the diameter of forming liposomes.

  15. Multiple star formation

    NASA Astrophysics Data System (ADS)

    Kraus, Adam L.

    2010-11-01

    In this thesis, I present a study of the formation and evolution of stars, particularly multiple stellar systems. Binary stars provide a key constraint on star formation because any successful model should reproduce the mass-dependent frequency, distribution of separations, and distribution of mass ratios. I have pursued a number of surveys for different ranges of parameter space, all yielding one overarching conclusion: binary formation is fundamentally tied to mass. Solar-mass stars have a high primordial binary frequency (50%--75%) and a wide range of separations (extending to >10,000 AU), but as the system mass decreases, the frequency and separation distribution also decrease. For brown dwarfs, binaries are rare (~10%--15%) and have separations of <5 AU. Inside of this outer separation cutoff, the separation distribution appears to be log-flat for solar-mass stars, and perhaps for lower-mass systems. Solar-mass binary systems appear to have a flat mass ratio distribution, but for primary masses <0.3 Msun, the distribution becomes increasingly biased toward similar-mass companions. My results also constrain the binary formation timescale and the postformation evolutionary processes that sculpt binary populations. The dynamical interaction timescale in sparse associations like Taurus and Upper Sco is far longer than their ages, which suggests that those populations are dynamically pristine. However, binary systems in denser clusters undergo significant dynamical processing that strips outer binary companions; the difference in wide binary properties between my sample and the field is explained by the composite origin of the field population. I also have placed the individual components of young binary systems on the HR diagram in order to infer their coevality. In Taurus, binary systems are significantly more coeval (Δτ~0.5 Myr) than the association as a whole (Δτ~3--5 Myr). Finally, my survey of young very-low-mass stars and brown dwarfs found no planetary

  16. Modeling river delta formation.

    PubMed

    Seybold, Hansjörg; Andrade, José S; Herrmann, Hans J

    2007-10-23

    A model to simulate the time evolution of river delta formation process is presented. It is based on the continuity equation for water and sediment flow and a phenomenological sedimentation/erosion law. Different delta types are reproduced by using different parameters and erosion rules. The structures of the calculated patterns are analyzed in space and time and compared with real data patterns. Furthermore, our model is capable of simulating the rich dynamics related to the switching of the mouth of the river delta. The simulation results are then compared with geological records for the Mississippi River. PMID:17940031

  17. Modeling river delta formation

    PubMed Central

    Seybold, Hansjörg; Andrade, José S.; Herrmann, Hans J.

    2007-01-01

    A model to simulate the time evolution of river delta formation process is presented. It is based on the continuity equation for water and sediment flow and a phenomenological sedimentation/erosion law. Different delta types are reproduced by using different parameters and erosion rules. The structures of the calculated patterns are analyzed in space and time and compared with real data patterns. Furthermore, our model is capable of simulating the rich dynamics related to the switching of the mouth of the river delta. The simulation results are then compared with geological records for the Mississippi River. PMID:17940031

  18. Adiabatic Halo Formation

    SciTech Connect

    Bazzani, A.; Turchetti, G.; Benedetti, C.; Rambaldi, S.; Servizi, G.

    2005-06-08

    In a high intensity circular accelerator the synchrotron dynamics introduces a slow modulation in the betatronic tune due to the space-charge tune depression. When the transverse motion is non-linear due to the presence of multipolar effects, resonance islands move in the phase space and change their amplitude. This effect introduces the trapping and detrapping phenomenon and a slow diffusion in the phase space. We apply the neo-adiabatic theory to describe this diffusion mechanism that can contribute to halo formation.

  19. Chemical analysis, inhibition of biofilm formation and biofilm eradication potential of Euphorbia hirta L. against clinical isolates and standard strains

    PubMed Central

    2013-01-01

    Background The frequent occurrences of antibiotic-resistant biofilm forming pathogens have become global issue since various measures that had been taken to curb the situation led to failure. Euphorbia hirta, is a well-known ethnomedicinal plant of Malaysia with diverse biological activities. This plant has been used widely in traditional medicine for the treatment of gastrointestinal, bronchial and respiratory ailments caused by infectious agents. Methods In the present study, chemical compositions of methanol extract of E. hirta L. aerial part was analyzed by gas chromatography and gas chromatography coupled to mass spectrometry. A relevant in vitro model was developed to assess the potency of the E. hirta extract to inhibit the bacterial biofilm formation as well as to eradicate the established biofilms. Besides biofilm, E. hirta extract was also evaluated for the inhibition efficacy on planktonic cells using tetrazolium microplate assay. For these purposes, a panel of clinically resistant pathogens and American type culture collection (ATCC) strains were used. Results The methanolic extract of aerial part of E. hirta was predominantly composed of terpenoid (60.5%) which is often regarded as an active entity accountable for the membrane destruction and biofilm cell detachment. The highest antibacterial effect of crude E. hirta extract was observed in the clinical isolates of Pseudomonas aeruginosa with minimum inhibitory concentration (MIC) value of 0.062 mg/ml. The extract also displayed potent biofilm inhibition and eradication activity against P. aeruginosa with minimum biofilm inhibition concentration (MBIC) and minimum biofilm eradication concentration (MBEC) values of 0.25 mg/ml and 0.5 mg/ml, respectively. Conclusions The crude methanol extract of E. hirta has proven to have interesting and potential anti-biofilm properties. The findings from this study will also help to establish a very promising anti-infective phytotherapeutical to be exploited in

  20. Star Formation in Irregular Galaxies.

    ERIC Educational Resources Information Center

    Hunter, Deidre; Wolff, Sidney

    1985-01-01

    Examines mechanisms of how stars are formed in irregular galaxies. Formation in giant irregular galaxies, formation in dwarf irregular galaxies, and comparisons with larger star-forming regions found in spiral galaxies are considered separately. (JN)

  1. Formats for Presenting Procedural Instruction

    ERIC Educational Resources Information Center

    Blaiwes, Arthur S.

    1974-01-01

    Thirty male college students performed a mock communication controller's task under three different instructional formats, short sentences, logical tree, and coding. It was concluded that format variations mainly influence the more difficult tasks. (Author/DE)

  2. Tooth formation - delayed or absent

    MedlinePlus

    Delayed or absent tooth formation; Teeth - delayed or absent formation ... The age at which the tooth comes in varies. Most infants get their first tooth between 6 and 9 months, but it may be earlier or later. ...

  3. Bubble formation in microgravity

    NASA Technical Reports Server (NTRS)

    Antar, Basil N.

    1994-01-01

    Two KC-135 flight campaigns have been conducted to date which are specifically dedicated to study bubble formation in microgravity. The first flight was conducted during March 14-18, 1994, and the other during June 20-24, 1994. The results from the June 1994 flight have not been analyzed yet, while the results from the March flight have been partially analyzed. In the first flight three different experiments were performed, one with the specific aim at determining whether or not cavitation can take place during any of the fluid handling procedures adopted in the shuttle bioprocessing experiments. The other experiments were concerned with duplicating some of the procedures that resulted in bubble formation, namely the NCS filling procedure and the needle scratch of a solid surface. The results from this set of experiments suggest that cavitation did not take place during any of the fluid handling procedures. The results clearly indicate that almost all were generated as a result of the breakup of the gas/liquid interface. This was convincingly demonstrated in the scratch tests as well as in the liquid fill tests.

  4. The Planet Formation Imager

    NASA Astrophysics Data System (ADS)

    Kraus, S.; Buscher, D. F.; Monnier, J. D.; PFI Science, the; Technical Working Group

    2014-04-01

    Among the most fascinating and hotly-debated areas in contemporary astrophysics are the means by which planetary systems are assembled from the large rotating disks of gas and dust which attend a stellar birth. Although important work is being done both in theory and observation, a full understanding of the physics of planet formation can only be achieved by opening observational windows able to directly witness the process in action. The key requirement is then to probe planet-forming systems at the natural spatial scales over which material is being assembled. By definition, this is the so-called Hill Sphere which delineates the region of influence of a gravitating body within its surrounding environment. The Planet Formation Imager project has crystallized around this challenging goal: to deliver resolved images of Hill-Sphere-sized structures within candidate planet-hosting disks in the nearest star-forming regions. In this contribution we outline the primary science case of PFI and discuss how PFI could significantly advance our understanding of the architecture and potential habitability of planetary systems. We present radiation-hydrodynamics simulations from which we derive preliminary specifications that guide the design of the facility. Finally, we give an overview about the interferometric and non-interferometric technologies that we are investigating in order to meet the specifications.

  5. Fullerene formation and annealing

    SciTech Connect

    Mintmire, J.W.

    1996-04-05

    Why does the highly symmetric carbon cluster C{sub 60} form in such profusion under the right conditions? This question was first asked in 1985, when Kroto suggested that the predominance of the C{sub 60} carbon clusters observed in the molecular beam experiments could be explained by the truncated icosahedral (or soccer ball) form. The name given to this cluster, buckminsterfullerene, led to the use of the term fullerenes for the family of hollow-cage carbon clusters made up of even numbers of triply coordinated carbons arranged with 12 pentagonal rings and an almost arbitrary number of hexagonal rings. More than a decade later, we still lack a completely satisfying understanding of the fundamental chemistry that takes place during fullerene formation. Most current models for fullerene formation require a facile mechanism for ring rearrangement in the fullerene structure, but the simplest proposed mechanisms are believed to have unrealistically high activation barriers. In recent research calculations have suggested that atomic carbon in the reaction mixture could act as a catalyst and allow substantially lower activation barriers for fullerene annealing. This article discusses the background for this research and other adjunct research. 14 refs.

  6. Biofilm formation by haloarchaea.

    PubMed

    Fröls, Sabrina; Dyall-Smith, Mike; Pfeifer, Felicitas

    2012-12-01

    A fluorescence-based live-cell adhesion assay was used to examine biofilm formation by 20 different haloarchaea, including species of Halobacterium, Haloferax and Halorubrum, as well as novel natural isolates from an Antarctic salt lake. Thirteen of the 20 tested strains significantly adhered (P-value  < 0.05) to a plastic surface. Examination of adherent cell layers on glass surfaces by differential interference contrast, fluorescence and confocal microscopy showed two types of biofilm structures. Carpet-like, multi-layered biofilms containing micro- and macrocolonies (up to 50 μm in height) were formed by strains of Halobacterium salinarum and the Antarctic isolate t-ADL strain DL24. The second type of biofilm, characterized by large aggregates of cells adhering to surfaces, was formed by Haloferax volcanii DSM 3757T and Halorubrum lacusprofundi DL28. Staining of the biofilms formed by the strongly adhesive haloarchaeal strains revealed the presence of extracellular polymers, such as eDNA and glycoconjugates, substances previously shown to stabilize bacterial biofilms. For Hbt. salinarum DSM 3754T and Hfx. volcanii DSM 3757T , cells adhered within 1 day of culture and remained viable for at least 2 months in mature biofilms. Adherent cells of Hbt. salinarum DSM 3754T showed several types of cellular appendages that could be involved in the initial attachment. Our results show that biofilm formation occurs in a surprisingly wide variety of haloarchaeal species. PMID:23057712

  7. STUDIES ON SHELL FORMATION

    PubMed Central

    Watabe, Norimitsu; Wilbur, Karl M.

    1961-01-01

    Details of crystal growth in the calcitostracum of Crassostrea virginica have been studied with the purpose of analyzing the formation of the overlapping rows of oriented tabular crystals characteristic of this part of the shell. Crystal elongation, orientation, and dendritic growth suggest the presence of strong concentration gradients in a thin layer of solution in which crystallization occurs. Formation of the overlapping rows can be explained by three processes observed in the shell: a two-dimensional tree-like dendritic growth in which one set of crystal branchings creeps over an adjacent set of branchings; three-dimensional dendritic growth; and growth by dislocation of crystal surfaces. Multilayers of crystals may thus be formed at one time. This is favored by infrequent secretion of a covering organic matrix which would inhibit crystal growth. The transitional zone covering the outer part of the calcitostracum and the inner part of the prismatic region is generally characterized by aggregates of small crystals with definite orientation. Growth in this zone appears to take place in a relatively homogeneous state of solution without strong concentration gradients. Thin membranes and bands of organic matrix were commonly observed in the transitional zone bordering the prismatic region. The membrane showed a very fine oriented network pattern. PMID:13783329

  8. Urbanization and Slum Formation

    PubMed Central

    Phua, Kai Hong

    2007-01-01

    The formation of slums need not be inevitable with rapid urbanization. Such an argument appears to be contradicted by evidence of large slum populations in a large number of developing countries and particularly in rapidly urbanizing regions like Asia. The evidence discussed suggests that city authorities faced with rapid urban development lack the capacity to cope with the diverse demands for infrastructural provision to meet economic and social needs. Not only are strategic planning and intervention major issues in agenda to manage rapid urbanization, but city governments are not effectively linking the economic development trajectory to implications for urban growth and, hence, housing needs. In the following discussion, a case study is presented in support of the argument that city governments have to first recognize and then act to establish the link that is crucial between economic development, urban growth, and housing. This is the agendum that has been largely neglected by city and national governments that have been narrowly focused on economic growth with the consequent proliferation of slum formation as a housing solution. PMID:17387618

  9. Modeling of microstructure formation

    SciTech Connect

    Rappaz, M.; Gandin, C.A.; Jacot, A.; Charbon, C.

    1995-12-31

    As macroscopic models of solidification are now well advanced, the simulation of microstructure formation is becoming increasingly important. Tools based on Greens` functions (i.e., front-tracking) or diffuse interface methods (e.g., phase field) have been developed recently for the calculation of individual dendritic grains or of a few eutectic lamellae. Although very powerful and useful, such methods cannot be extended at present to the scale of a whole process mainly because of the very large computation time involved. At the intermediate mesoscopic scale of the grains, Monte Carlo (MC) or Cellular Automata (CA) methods can integrate nucleation and grain growth mechanisms in order to simulate the formation of grains during solidification. These latter methods have been coupled with Finite Element (FE) heat flow calculations in order to predict the grain structure at the scale of a whole process (computer metallography). The microstructural features which can be predicted using this coupled CA-FE model are: the morphology of the grains (columnar, equiaxed), the columnar-to-equiaxed transition, the selection of grains in the columnar zone, the crystallographic texture of the grains, the extension of grains in open regions of liquid, etc. Calculated parameters of the three-dimensional grain structure can also be related to the same entities obtained in metallographic cross sections (computer stereology).

  10. Method for measuring pollutant formation

    NASA Technical Reports Server (NTRS)

    Annen, Kurt (Inventor); Stickler, David B. (Inventor)

    2001-01-01

    Diagnostic methods for determining an instantaneous rate of pollutant formation in a combustion system are based on measurement of chemiluminescence intensity generated simultaneously with the formation of the pollutant. The chemiluminescent signal is generated by an analog reaction which occurs in parallel with a key step in the formation of a specific pollutant of interest. The connection between the analog reaction and the pollution reaction is such that the chemiluminescent signal indicates the local, instantaneous formation rate of the pollutant of interest.

  11. Iron limitation enhances acyl homoserine lactone (AHL) production and biofilm formation in clinical isolates of Acinetobacter baumannii

    PubMed Central

    Modarresi, Farzan; Azizi, Omid; Shakibaie, Mohammad Reza; Motamedifar, Mohammad; Mosadegh, Ellahe; Mansouri, Shahla

    2015-01-01

    the biofilm formation introduced 23 isolates with efficient attachment to microplate wells and strong biofilm. We found that both the AHL production and biofilm formation were regulated by iron concentration in a dose dependent manner. These findings provide evidence that iron limitation plays an important regulatory role in AHL and siderophore production resulting in strong or weak biofilm, thereby helping the organism to persist in less available micronutrient environment. PMID:25622119

  12. Formative Assessment: Simply, No Additives

    ERIC Educational Resources Information Center

    Roskos, Kathleen; Neuman, Susan B.

    2012-01-01

    Among the types of assessment the closest to daily reading instruction is formative assessment. In contrast to summative assessment, which occurs after instruction, formative assessment involves forming judgments frequently in the flow of instruction. Key features of formative assessment include identifying gaps between where students are and…

  13. LISA satellite formation control

    NASA Astrophysics Data System (ADS)

    Bik, J. J. C. M.; Visser, P. N. A. M.; Jennrich, O.

    The joint ESA-NASA Laser Interferometer Space Antenna (LISA) mission consists of a triangular formation of three satellites aiming at detecting gravitational waves. In linear approximation the LISA satellites describe a circle around a reference point, maintaining a fixed position with respect to each other. The reference point, the center of the triangle, orbits the Sun in a circular orbit, trailing the Earth at twenty degrees. In reality the distance between the satellites will vary by about one to two percent and the angle between the arms of the antenna will vary by about 0.5° over the course of one year for the nominal LISA satellite configuration. For measurement accuracy it is desirable that the pointing offset of the telescopes be kept small. This makes it necessary to actuate the telescopes or to control the formation. It was assumed that the LISA satellites are equipped with six μN engines that would allow to keep the two cubical proof masses within each satellite in almost perfect free fall. It was found that control forces up to about 700 μN are required for maintaining the absolute triangular LISA formation, leading to unacceptable excursions of the proof masses from free fall. However, these forces compensate predominantly very low frequency variations of the arm lengths and angles of the triangle, which are then to be compensated by the telescope actuators. The variations are outside the aimed LISA measurement bandwidth (10 -4-0.1 Hz). In addition, the effect of thruster noise, orbit determination errors and orbit injection errors was examined. The effect of these error sources on the arm lengths and orientation angles between the LISA satellites was assessed both in open loop and in closed loop, where the closed loop was based on a proportional-derivative (PD) controller. It was found that orbit determination errors of the order of a few km in position and a few mm/s in velocity lead to negligible closed loop control forces. In addition, orbit

  14. Theories of galaxy formation

    SciTech Connect

    Jones, B.J.T.

    1980-01-01

    The current status of some theories of galaxy formation that are consistent with the hot big bang origin of the universe is reviewed. In the cosmic turbulence theory, an attempt is made to explain not only the characteristic masses and angular momenta of galaxies, but to describe in detail the spectrum of galaxy clustering problems with regard to the observed abundances of the light elements, a Kolmogorov spectrum of turbulence and the fireball are discussed. Attention is given to a primordial chaotic magnetic field, the comparison between baryon-symmetric cosmologies, the origin of galactic spin and theories starting from isothermal perturbations. Also considered are the dilemma of the initial conditions with respect to the era after 10 to the -4th s, and the pancake theory, in which the planar structures that arise provide a natural explanation for filamentary structures.

  15. Physics of amniote formation.

    PubMed

    Fleury, Vincent; Murukutla, Ameya Vaishnavi; Chevalier, Nicolas R; Gallois, Benjamin; Capellazzi-Resta, Marina; Picquet, Pierre; Peaucelle, Alexis

    2016-08-01

    We present a detailed study of the formation of the amniotic sac in the avian embryo, and a comparison with the crocodile amniotic sac. We show that the amniotic sac forms at a circular line of stiffness contrast, separating rings of cell domains. Cells align at this boundary, and this in turn orients and concentrates the tension forces. The tissue fold which forms the amniotic sac is locked exactly along this line due to the colocalization of the stiffness contrast and of the tensile force. In addition, the tensile force plays a regenerative role when the amniotic sac is cut. The fold forming the ventral side of the embryo displays the same characteristics. This work shows that amniote embryogenesis consists of a cascade of buckling events taking place at the boundaries between regions of differing mechanical properties. Hence, amniote embryogenesis relies on a simple and robust biomechanical scheme used repeatedly, and selected ancestrally. PMID:27627351

  16. Pattern Formation in Materials

    NASA Astrophysics Data System (ADS)

    Karma, Alain

    2011-04-01

    Pattern formation is ubiquitous in nature, from sand ripples formed by wind to the development of a complex biological organism with different organs and a central nervous system. In the realm of materials, patterns are formed invariably when matter is transformed between different solid, liquid or gaseous states far from thermodynamic equilibrium. Material failure is itself mediated by the propagation of cracks that form intricate patterns. Understanding how patterns form and evolve is key to design materials with desired properties and to optimize their performance and safety. This talk will discuss recent progress made to understand three distinct class of patterns including the highly branched snow-flake-like dendritic patterns formed during the solidification process, polycrystalline patterns shaped by grain boundaries, and crack patterns.

  17. Dislocation Formation in Alloys

    NASA Astrophysics Data System (ADS)

    Minami, Akihiko; Onuki, Akira

    2006-05-01

    An interaction between dislocations and phase transitions is studied by a phase field model both in two and three dimensional systems. Our theory is a simple extension of the traditional linear elastic theory, and the elastic energy is a periodic function of local strains which is reflecting the periodicity of crystals. We find that the dislocations are spontaneously formed by quenching. Dislocations are formed from the interface of binary alloys, and slips are preferentially gliding into the soft metals. In three dimensional systems, formation of dislocations under applied strain is studied in two phase state. We find that the dislocation loops are created from the surface of hard metals. We also studied the phase separation above the coexisting temperature which is called as the Cottrell atmosphere. Clouds of metals cannot catch up with the motion of dislocations at highly strained state.

  18. Group Formation in Economics

    NASA Astrophysics Data System (ADS)

    Demange, Gabrielle; Wooders, Myrna

    2005-01-01

    Broad and diverse ranges of activities are conducted within and by organized groups of individuals, including political, economic and social activities. These activities have recently become a subject of intense interest in economics and game theory. Some of the topics investigated in this collection are models of networks of power and privilege, trade networks, co-authorship networks, buyer-seller networks with differentiated products, and networks of medical innovation and the adaptation of new information. Other topics are social norms on punctuality, clubs and the provision of club goods and public goods, research and development and collusive alliances among corporations, and international alliances and trading agreements. While relatively recent, the literature on game theoretic studies of group formation in economics is already vast. This volume provides an introduction to this important literature on game-theoretic treatments of situations with networks, clubs, and coalitions, including some applications.

  19. Mechanism of GEMS formation

    SciTech Connect

    Bradley, J P; Dai, Z R

    2004-03-10

    GEMS (glass with embedded metal and sulfides) in interplanetary dust particles (IDPs) were examined using 200 keV analytical transmission electron microscopy. The morphologies and crystallography of embedded relict grains reveal that GEMS are pseudomorphs formed by irradiation processing of crystals free-floating in space. Some GEMS retain a compositional and morphological ''memory'' of the crystal from which they formed. Pseudomorphism rules out condensation, annealing, flash heating, or shock melting as alternative mechanisms of GEMS formation. A significant and often dominant fraction of the atoms in GEMS were sputtered deposited from other grains. Therefore, a normal (solar) isotopic composition is not a reliable indicator of whether GEMS formed in the solar system or in presolar interstellar or circumstellar environments.

  20. Myxobacteria Fruiting Body Formation

    NASA Astrophysics Data System (ADS)

    Jiang, Yi

    2006-03-01

    Myxobacteria are social bacteria that swarm and glide on surfaces, and feed cooperatively. When starved, tens of thousands of cells change their movement pattern from outward spreading to inward concentration; they form aggregates that become fruiting bodies, inside which cells differentiate into nonmotile, environmentally resistant spores. Traditionally, cell aggregation has been considered to imply chemotaxis, a long-range cell interaction mediated by diffusing chemicals. However, myxobacteria aggregation is the consequence of direct cell-contact interactions. I will review our recent efforts in modeling the fruiting body formation of Myxobacteria, using lattice gas cellular automata models that are based on local cell-cell contact signaling. These models have reproduced the individual phases in Myxobacteria development such as the rippling, streaming, early aggregation and the final sporulation; the models can be unified to simulate the whole developmental process of Myxobacteria.

  1. DUST FORMATION IN MACRONOVAE

    SciTech Connect

    Takami, Hajime; Ioka, Kunihito; Nozawa, Takaya E-mail: kunihito.ioka@kek.jp

    2014-07-01

    We examine dust formation in macronovae (as known as kilonovae), which are the bright ejecta of neutron star binary mergers and one of the leading sites of r-process nucleosynthesis. In light of information about the first macronova candidate associated with GRB 130603B, we find that dust grains of r-process elements have difficulty forming because of the low number density of the r-process atoms, while carbon or elements lighter than iron can condense into dust if they are abundant. Dust grains absorb emission from ejecta with an opacity even greater than that of the r-process elements, and re-emit photons at infrared wavelengths. Such dust emission can potentially account for macronovae without r-process nucleosynthesis as an alternative model. This dust scenario predicts a spectrum with fewer features than the r-process model and day-scale optical-to-ultraviolet emission.

  2. Glass formation in microgravity

    NASA Technical Reports Server (NTRS)

    Ray, C. S.; Day, D. E.

    1987-01-01

    An account is given of containerless glass-forming experiments conducted aboard the Space Shuttle in 1985, using a single-axis acoustic levitator furnace apparatus. An attempt was made to obtain quantitative evidence for the suppression of heterogeneous nucleation/crystallization in containerless melts under microgravity conditions, as well as to study melt homogenization in the absence of gravity-driven convection and assess the feasibility of laser fusion target glass microsphere preparation with a microgravity apparatus of the present type. A ternary calcia-gallia-silica glass thus obtained indicated a 2-3-fold increase in glass-formation tendency for this material composition in microgravity, by comparison with 1g.

  3. Recipes for planet formation

    NASA Astrophysics Data System (ADS)

    Meyer, Michael R.

    2009-11-01

    Anyone who has ever used baking soda instead of baking powder when trying to make a cake knows a simple truth: ingredients matter. The same is true for planet formation. Planets are made from the materials that coalesce in a rotating disk around young stars - essentially the "leftovers" from when the stars themselves formed through the gravitational collapse of rotating clouds of gas and dust. The planet-making disk should therefore initially have the same gas-to-dust ratio as the interstellar medium: about 100 to 1, by mass. Similarly, it seems logical that the elemental composition of the disk should match that of the star, reflecting the initial conditions at that particular spot in the galaxy.

  4. Microplate model for the present-day deformation of Tibet

    USGS Publications Warehouse

    Thatcher, W.

    2007-01-01

    Site velocities from 349 Global Positioning System (GPS) stations are used to construct an 11-element quasi-rigid block model of the Tibetan Plateau and its surroundings. Rigid rotations of five major blocks are well determined, and average translation velocities of six smaller blocks can be constrained. Where data are well distributed the velocity field can be explained well by rigid block motion and fault slip across block boundaries. Residual misfits average 1.6 mm/yr compared to typical one standard deviation velocity uncertainties of 1.3 mm/yr. Any residual internal straining of the blocks is small and heterogeneous. However, residual substructure might well represent currently unresolved motions of smaller blocks. Although any smaller blocks must move at nearly the same rate as the larger blocks within which they lie, undetected relative motions between them could be significant, particularly where there are gaps in GPS coverage. Predicted relative motions between major blocks agree with the observed sense of slip and along-strike partitioning of motion across major faults. However, predicted slip rates across Tibet's major strike-slip faults are low, only 5-12 mm/yr, a factor of 2-3 smaller than most rates estimated from fault offset features dated by radiometric methods as ???2000 to ???100,000 year old. Previous work has suggested that both GPS data and low fault slip rates are incompatible with rigid block motions of Tibet. The results reported here overcome these objections.

  5. Quantitative Microplate Assay for Real-Time Nuclease Kinetics

    PubMed Central

    Langel, Ülo

    2016-01-01

    Utilizing the phenomenon of nucleases exposing oligonucleotide phosphate backbones to phosphatases we present a novel quantitative method for kinetics of nuclease catalysis. Inorganic phosphate released from nuclease products by phosphatases could be quantified in real-time by a fluorescent sensor of inorganic phosphate. Two different nucleases were employed, showing the versatility of this assay for multiple turnover label-free nuclease studies. PMID:27101307

  6. Streamer formation in sprites

    NASA Astrophysics Data System (ADS)

    McHarg, M. G.; Kammae, T.; Nielsen, H. C.

    2005-12-01

    Models of sprite formation for positive cloud-to-ground lightning strokes predict both downward (positive), and upward (negative) propagating streamers. Previous high speed camera observations of sprites are generally consistent with these predictions, but have been unable to resolve the temporal formation of the streamers due to frame rates limited to a few thousand frames per second. We report observations made during the evening of 9 July 2005 at 10,000 frames per second, with the image intensifier gated to 50 microseconds per frame. These observations often show the streamer head to be a bead-like structure propagating downward at approximately 7x106 m/s for 1,500 microseconds. The bead is followed by a dark region, and the main emissions from the sprite column are delayed ~800 microseconds after the passage of the streamer head. There are also "beads" which clearly propagate upward. Some events appear to be very similar to laboratory images of time resolved streamer zones. We interpret these observations in terms of positive/negative streamers. We see evidence for branching of the streamer tips in several cases, as well as evidence of upward propagating streamers transitioning into a more diffuse emission. Previous work (Pasko and Stenbaek-Nielsen, GRL 29(10), 2002) indicates this transition region has a lower border at an altitude when the dielectric relaxation time scale equals the time scale for an individual electron to develop into a streamer, and an upper border when the dielectric relaxation time scale roughly equals the dissociative attachment time scale. The present observations appear to be broadly consistent with this interpretation.

  7. Hazards to Planet Formation

    NASA Astrophysics Data System (ADS)

    Bally, J.

    2001-05-01

    The Orion Nebula provides a remarkable window on the first few million years in the lives of typical young stars and planetary systems. HST has demonstrated that most young stars in the Nebula are surrounded by circumstellar disks (the so-called `proplyds'). While these observations show that planet forming environments may be common, they also demonstrate that Orion's disks are being destroyed by intense UV radiation fields. `Gravel' sufficiently large to resist photo-erosion (meter scale solids or ices) may lock-up sufficient material to eventually build rocky planets. Indeed, there is evidence for large solids in some proplyds. But, the hydrogen and helium needed for the formation of giant planets will be removed. To form in Orion-like environments, giant planets must be assembled promptly prior to UV exposure. Even rocky planets may not form if the photoionized disk corona causes surviving large particles in the disk to spiral into the central star. Thus, nearby massive stars pose severe hazards to planet formation. Star counts indicate that most stars form in Orion-like environments. Only about 10% of young stars are born in shielded environments such as the Taurus or L1641 clouds where disks may escape photo-erosion. In dark clouds, the majority of stars (> 80%) form in non-hierarchal multiple star systems where close encounters with sibling stars can destroy disks and eject young planets. Thus, most stars may never develop planetary systems. These considerations indicate that extra-Solar planets may be rare, contrary to the popular view. These conclusions are consistent with the recent discoveries of extra-Solar planets around a few percent of single stars.

  8. Medusae Fossae Formation

    NASA Technical Reports Server (NTRS)

    2002-01-01

    (Released 16 April 2002) The Science This THEMIS visible image was acquired near 11o N, 159o W (201o E) and shows examples of the remarkable variations that can be seen in the erosion of the Medusae Fossae Formation. This Formation is a soft, easily eroded deposit that extends for nearly 1,000 km along the equator of Mars. In this region, like many others throughout the Medusae Fossae Formation, the surface has been eroded by the wind into a series of linear ridges called yardangs. These ridges generally point in direction of the prevailing winds that carved them, and demonstrate the power of martian winds to erode the landscape of Mars. The easily eroded nature of the Medusae Fossae Formation suggests that it is composed of weakly cemented particles, and was most likely formed by the deposition of wind-blown dust or volcanic ash. Within this single image it is possible to see differing amounts of erosion and stripping of layers in the Medusae Fossae Formation. Near the bottom (southern) edge of the image a rock layer with a relatively smooth upper surface covers much of the image. Moving upwards (north) in the image this layer becomes more and more eroded. At first there are isolated regions where the smooth unit has been eroded to produce sets of parallel ridges and knobs. Further north these linear knobs increase in number, and only small, isolated patches of the smooth upper surface remain. Finally, at the top of the image, even the ridges have been removed, exposing the remarkably smooth top of hard, resistant layer below. This sequence of layers with differing hardness and resistance to erosion is common on Earth and on Mars, and suggests significant variations in the physical properties, composition, particle size, and/or cementation of these martian layers. As is common throughout the Medusae Fossae Formation, very few impact craters are visible, indicating that the surface exposed is relatively young, and that the process of erosion may be active today

  9. Medusae Fossae Formation

    NASA Technical Reports Server (NTRS)

    2002-01-01

    (Released 16 April 2002) The Science This THEMIS visible image was acquired near 11o N, 159o W (201o E) and shows examples of the remarkable variations that can be seen in the erosion of the Medusae Fossae Formation. This Formation is a soft, easily eroded deposit that extends for nearly 1,000 km along the equator of Mars. In this region, like many others throughout the Medusae Fossae Formation, the surface has been eroded by the wind into a series of linear ridges called yardangs. These ridges generally point in direction of the prevailing winds that carved them, and demonstrate the power of martian winds to erode the landscape of Mars. The easily eroded nature of the Medusae Fossae Formation suggests that it is composed of weakly cemented particles, and was most likely formed by the deposition of wind-blown dust or volcanic ash. Within this single image it is possible to see differing amounts of erosion and stripping of layers in the Medusae Fossae Formation. Near the bottom (southern) edge of the image a rock layer with a relatively smooth upper surface covers much of the image. Moving upwards (north) in the image this layer becomes more and more eroded. At first there are isolated regions where the smooth unit has been eroded to produce sets of parallel ridges and knobs. Further north these linear knobs increase in number, and only small, isolated patches of the smooth upper surface remain. Finally, at the top of the image, even the ridges have been removed, exposing the remarkably smooth top of hard, resistant layer below. This sequence of layers with differing hardness and resistance to erosion is common on Earth and on Mars, and suggests significant variations in the physical properties, composition, particle size, and/or cementation of these martian layers. As is common throughout the Medusae Fossae Formation, very few impact craters are visible, indicating that the surface exposed is relatively young, and that the process of erosion may be active today

  10. Evaluation of a High-Throughput Peptide Reactivity Format Assay for Assessment of the Skin Sensitization Potential of Chemicals

    PubMed Central

    Wong, Chin Lin; Lam, Ai-Leen; Smith, Maree T.; Ghassabian, Sussan

    2016-01-01

    The direct peptide reactivity assay (DPRA) is a validated method for in vitro assessment of the skin sensitization potential of chemicals. In the present work, we describe a peptide reactivity assay using 96-well plate format and systematically identified the optimal assay conditions for accurate and reproducible classification of chemicals with known sensitizing capacity. The aim of the research is to ensure that the analytical component of the peptide reactivity assay is robust, accurate, and reproducible in accordance with criteria that are used for the validation of bioanalytical methods. Analytical performance was evaluated using quality control samples (QCs; heptapeptides at low, medium, and high concentrations) and incubation of control chemicals (chemicals with known sensitization capacity, weak, moderate, strong, extreme, and non-sensitizers) with each of three synthetic heptapeptides, viz Cor1-C420 (Ac-NKKCDLF), cysteine- (Ac-RFAACAA), and lysine- (Ac-RFAAKAA) containing heptapeptides. The optimal incubation temperature for all three heptapeptides was 25°C. Apparent heptapeptide depletion was affected by vial material composition. Incubation of test chemicals with Cor1-C420, showed that peptide depletion was unchanged in polypropylene vials over 3-days storage in an autosampler but this was not the case for borosilicate glass vials. For cysteine-containing heptapeptide, the concentration was not stable by day 3 post-incubation in borosilicate glass vials. Although the lysine-containing heptapeptide concentration was unchanged in both polypropylene and borosilicate glass vials, the apparent extent of lysine-containing heptapeptide depletion by ethyl acrylate, differed between polypropylene (24.7%) and glass (47.3%) vials. Additionally, the peptide-chemical complexes for Cor1-C420-cinnamaldehyde and cysteine-containing heptapeptide-2, 4-dinitrochlorobenzene were partially reversible during 3-days of autosampler storage. These observations further highlight

  11. Gas formation. Formation temperatures of thermogenic and biogenic methane.

    PubMed

    Stolper, D A; Lawson, M; Davis, C L; Ferreira, A A; Santos Neto, E V; Ellis, G S; Lewan, M D; Martini, A M; Tang, Y; Schoell, M; Sessions, A L; Eiler, J M

    2014-06-27

    Methane is an important greenhouse gas and energy resource generated dominantly by methanogens at low temperatures and through the breakdown of organic molecules at high temperatures. However, methane-formation temperatures in nature are often poorly constrained. We measured formation temperatures of thermogenic and biogenic methane using a "clumped isotope" technique. Thermogenic gases yield formation temperatures between 157° and 221°C, within the nominal gas window, and biogenic gases yield formation temperatures consistent with their comparatively lower-temperature formational environments (<50°C). In systems where gases have migrated and other proxies for gas-generation temperature yield ambiguous results, methane clumped-isotope temperatures distinguish among and allow for independent tests of possible gas-formation models. PMID:24970083

  12. Bubble formation in microgravity

    NASA Technical Reports Server (NTRS)

    Antar, Basil N.

    1996-01-01

    An extensive experimental program was initiated for the purpose of understanding the mechanisms leading to bubble generation during fluid handling procedures in a microgravity environment. Several key fluid handling procedures typical for PCG experiments were identified for analysis in that program. Experiments were designed to specifically understand how such procedures can lead to bubble formation. The experiments were then conducted aboard the NASA KC-135 aircraft which is capable of simulating a low gravity environment by executing a parabolic flight attitude. However, such a flight attitude can only provide a low gravity environment of approximately 10-2go for a maximum period of 30 seconds. Thus all of the tests conducted for these experiments were designed to last no longer than 20 seconds. Several experiments were designed to simulate some of the more relevant fluid handling procedures during protein crystal growth experiments. These include submerged liquid jet cavitation, filling of a cubical vessel, submerged surface scratch, attached drop growth, liquid jet impingement, and geysering experiments. To date, four separate KC-135 flight campaigns were undertaken specifically for performing these experiments. However, different experiments were performed on different flights.

  13. Nuclear ``pasta'' formation

    NASA Astrophysics Data System (ADS)

    Schneider, A. S.; Horowitz, C. J.; Hughto, J.; Berry, D. K.

    2013-12-01

    The formation of complex nonuniform phases of nuclear matter, known as nuclear pasta, is studied with molecular dynamics (MD) simulations containing 51200 nucleons. A phenomenological nuclear interaction is used that reproduces the saturation binding energy and density of nuclear matter. Systems are prepared at an initial density of 0.10fm-3 and then the density is decreased by expanding the simulation volume at different rates to densities of 0.01fm-3 or less. An originally uniform system of nuclear matter is observed to form spherical bubbles (“swiss cheese”), hollow tubes, flat plates (“lasagna”), thin rods (“spaghetti”) and, finally, nearly spherical nuclei with decreasing density. We explicitly observe nucleation mechanisms, with decreasing density, for these different pasta phase transitions. Topological quantities known as Minkowski functionals are obtained to characterize the pasta shapes. Different pasta shapes are observed depending on the expansion rate. This indicates nonequilibrium effects. We use this to determine the best ways to obtain lower energy states of the pasta system from MD simulations and to place constraints on the equilibration time of the system.

  14. Acromioclavicular joint cyst formation.

    PubMed

    Hiller, Andrew D; Miller, Joshua D; Zeller, John L

    2010-03-01

    Acromioclavicular joint (ACJ) cysts are an uncommon and unusual sequela associated with shoulder pathophysiology. The majority of literature on ACJ cysts consists of individual case reports with no definitive literature review currently available. In addition to a comprehensive literature review, four clinical cases are presented in this report. First described by Craig (1984), a total of 41 cases have been previously reported in the literature. Of these cases, five occurred with the rotator cuff musculature intact. The remaining 36 cases of ACJ cysts occurred in patients with a complete tear/avulsion of the rotator cuff. Previous attempts at compiling a complete record of all reported cases have combined several distinct conditions into a single category. This article presents two distinct etiologies for the pathogenesis of ACJ cyst formation. In the presence of an intact rotator cuff, a Type 1 cyst can form superficially and be limited to the ACJ. Following a massive or traumatic tear of the rotator cuff, mechanical instability of the humeral head can cause a deterioration of the inferior acromioclavicular capsule (cuff tear arthropathy) and an overproduction of synovial fluid. Overtime, a "geyser" of fluid can form between the glenohumeral and the ACJ, forming a Type 2 cyst. This differentiation and categorization is essential for appropriate classification and treatment. PMID:20069645

  15. Studies on Shell Formation

    PubMed Central

    Watabe, Norimitsu; Sharp, D. Gordon; Wilbur, Karl M.

    1958-01-01

    Electron microscope observations have been made by means of the replica method on growth processes of calcite crystals of the nacreous layer of the shell of the oyster, Crassostrea virginica. Layer formation is initiated by the secretion of a conchiolin matrix and the deposition of rounded crystal seeds on or in this material. In some areas crystal seeds are elongate and within a given area show a similar orientation, probably due to slower deposition. The seeds appear to increase in size by dendritic growth, and smaller seeds become incorporated into larger ones which come into contact to form a single layer. With further growth, crystals overlap, forming a step-like arrangement. The direction of growth is frequently different in neighboring regions. Crystal seeds deposited on crystal surfaces are usually elongate and oriented. Well developed crystals have a tabular idiomorphic form and are parallel in their growth. Rounded and irregular crystals were also observed. The crystals show reticular structure with units of the order of 100 A and striations corresponding with the rhombohedral axes of the crystals. The role of the mantle is discussed in relation to the growth patterns of crystals and shell structure. PMID:13549499

  16. Beach-cusp formation

    USGS Publications Warehouse

    Sallenger, A.H., Jr.

    1979-01-01

    Field experiments on beach-cusp formation were undertaken to document how the cuspate form develops and to test the edge-wave hypothesis on the uniform spacing of cusps. These involved observations of cusps forming from an initially plane foreshore. The cuspate form was observed to be a product of swash modification of an intertidal beach ridge as follows. A ridge, cut by a series of channels quasi-equally spaced along its length, was deposited onto the lower foreshore. The ridge migrated shoreward with flood tide, while the longshore positions of the channels remained fixed. On ebb tide, changes in swash circulation over the ridge allowed the upwash to flow shoreward through the channels and the channel mouths were eroded progressively wider until adjacent mouths met, effecting a cuspate shape. Measured spacings of cusps, ranging in size from less than 1 m to more than 12 m, agree well with computed spacings due to either zero-mode subharmonic or zero-mode synchronous edge waves. Edge-wave-induced longshore variations in run up will cause water ponded behind a ridge to converge at points of low swash and flow seaward as relatively narrow currents eroding channels spaced at one edge-wave wavelength for synchronous edge waves or one half wavelength for subharmonic edge waves. The channels are subsequently modified into cusp troughs as described above.

  17. Large Format Radiographic Imaging

    SciTech Connect

    J. S. Rohrer; Lacey Stewart; M. D. Wilke; N. S. King; S. A Baker; Wilfred Lewis

    1999-08-01

    Radiographic imaging continues to be a key diagnostic in many areas at Los Alamos National Laboratory (LANL). Radiographic recording systems have taken on many form, from high repetition-rate, gated systems to film recording and storage phosphors. Some systems are designed for synchronization to an accelerator while others may be single shot or may record a frame sequence in a dynamic radiography experiment. While film recording remains a reliable standby in the radiographic community, there is growing interest in investigating electronic recording for many applications. The advantages of real time access to remote data acquisition are highly attractive. Cooled CCD camera systems are capable of providing greater sensitivity with improved signal-to-noise ratio. This paper begins with a review of performance characteristics of the Bechtel Nevada large format imaging system, a gated system capable of viewing scintillators up to 300 mm in diameter. We then examine configuration alternatives in lens coupled and fiber optically coupled electro-optical recording systems. Areas of investigation include tradeoffs between fiber optic and lens coupling, methods of image magnification, and spectral matching from scintillator to CCD camera. Key performance features discussed include field of view, resolution, sensitivity, dynamic range, and system noise characteristics.

  18. The formation of dew

    NASA Astrophysics Data System (ADS)

    Beysens, D.

    Dew is the condensation into liquid droplets of water vapor on a substrate. The presence of a substrate is the origin of the peculiarities and richness of the phenomenon. We review the aspects related to heterogeneous nucleation and subsequent growth of water droplets. A key point is the drop interaction through drop fusion or coalescence, which leads to scaling in the growth and gives universality to the process. The effects of substrate heterogeneity and gravity effects are also considered. Coalescence events lead to temporal and spatio-temporal fluctuations in the substrate coverage, drop configuration, etc., which give rise to a very peculiar dynamics. When the substrate is a liquid or a liquid crystal, the drop pattern can exhibit special spatial orders, such as crystalline, hexatic phases and fractal contours. And condensation on a solid substrate near its melting point can make the drop jump. The applications of monitoring dew formation are manyfold. Examples can be found in nanoelectronics and optics (vapor deposition and thin films), medicine (sterilization process), agriculture (green houses). We here discuss in greater details the production of clean water by "atmospheric wells".

  19. Gaining Insight into Star Formation: Resolved Star Formation Laws

    NASA Astrophysics Data System (ADS)

    Liebst, Kelley; Scowen, Paul A.

    2014-06-01

    Until recently astronomers have used star formation laws to measure the star formation rate and star formation efficiency of galaxies only on global scales because of the poor resolution of available data. What I am now capable of producing is a spatially resolved star formation law that can provide direct insight into the physical processes that govern star formation and assess the short-term nature of bursts of star formation and the longer-term nature of larger-scale events that can dictate the global distribution of stars and the ultimate fate of a galaxy as a whole. I am using exquisite narrowband optical data from a variety of sources, including the Hubble Space Telescope, and Kitt Peak National Observatory, etc., in conjunction with infrared data from the Spitzer Infrared Nearby Galaxy Survey and the Spitzer Local Volume Legacy survey, neutral gas data from The HI Nearby Galaxy Survey, and molecular gas data from the Berkeley-Illinois-Maryland Association Survey of Nearby Galaxies, to provide star formation rates and star formation efficiencies on previously inaccessible small spatial scales across a suite of galaxies that represent a range of star formation environments and scales. My sample includes 18 spiral galaxies ranging from 2.1 to 15.1 Mpc in distance and offers a large range of morphological types (i.e. a large range of star formation environments). I am using these data to test different models of star formation modes under a variety of physical conditions and relate the variations I observe to the known local physical conditions and the associated star formation histories for each locale within each galaxy.This is the heart of the matter - that the nature and evolution of the local physical environment intimately influences how stars can form, how quickly and how massive those stars are allowed to form, and as a result how they shape the local conditions for subsequent star formation. It is this tracking of the stellar ecology that is vital for

  20. Formation of Bidisperse Particle Clouds

    NASA Astrophysics Data System (ADS)

    Er, Jenn Wei; Zhao, Bing; Law, Adrian W. K.; Adams, E. Eric

    2014-11-01

    When a group of dense particles is released instantaneously into water, their motion has been conceptualized as a circulating particle thermal (Ruggerber 2000). However, Wen and Nacamuli (1996) observed the formation of particle clumps characterized by a narrow, fast moving core shedding particles into wakes. They observed the clump formation even for particles in the non-cohesive range as long as the source Rayleigh number was large (Ra > 1E3) or equivalently the source cloud number (Nc) was small (Nc < 3.2E2). This physical phenomenon has been investigated by Zhao et al. (2014) through physical experiments. They proposed the theoretical support for Nc dependence and categorized the formation processes into cloud formation, transitional regime and clump formation. Previous works focused mainly on the behavior of monodisperse particles. The present study further extends the experimental investigation to the formation process of bidisperse particles. Experiments are conducted in a glass tank with a water depth of 90 cm. Finite amounts of sediments with various weight proportions between coarser and finer particles are released from a cylindrical tube. The Nc being tested ranges from 6E-3 to 9.9E-2, which covers all the three formation regimes. The experimental results showed that the introduction of coarse particles promotes cloud formation and reduce the losses of finer particles into the wake. More quantitative descriptions of the effects of source conditions on the formation processes will be presented during the conference.

  1. Plasmapause formation at Saturn

    NASA Astrophysics Data System (ADS)

    Thomsen, M. F.; Mitchell, D. G.; Jia, X.; Jackman, C. M.; Hospodarsky, G.; Coates, A. J.

    2015-04-01

    Cassini observations during a rapid, high-latitude, dawnside pass from Saturn's lobe to inner magnetosphere on 25 June 2009 provide strong evidence for the formation of a "plasmapause" at Saturn by Vasyliunas-type nightside reconnection of previously mass-loaded flux tubes. A population of hot, tenuous plasma that lies between the lobe and the dense inner magnetospheric plasma is consistent with a region formed by very recent injection from a reconnection region in the tail, including low density, high temperature, supercorotational flow, a significant O+ content, and the near-simultaneous observation of enhanced Saturn kilometric radiation emissions. The sharp boundary between that region and the cool dense inner magnetospheric plasma thus separates flux tubes that were involved in the reconnection from those that successfully traversed the nightside without mass loss. This event demonstrates that tail reconnection can strip off inner magnetospheric plasma in to at least dipole L = 8.6. Clear evidence of flux tube interchange driven by the sharp boundary is found, both inward moving flux tubes of hotter plasma and, for the first time, the outward moving cool population. The outward moving cool regions have azimuthal sizes less than 1 RS, were probably created within the past 1.2 h, and have outflow speeds greater than about 5 km/s. At the outer edge of the reconnected region, there is also a possible signature of Dungey-type lobe reconnection following the initial Vasyliunas-type reconnection. Observations from this event are entirely consistent with previously described global MHD simulations of tail reconnection, plasmoid departure, and Saturnward injection of reconnected flux.

  2. The Tuscaloosa Formation revisited

    SciTech Connect

    Hansley, P.L.

    1996-09-01

    A petrologic study of the Upper Cretaceous lower Tuscaloosa Formation in the Gulf Coast from depths of 2,700 to 6,000 in indicates that anomalously high porosity (>20 percent) in deep gas and condensate-bearing sandstones (5,000 to 6,000 m) is approximately evenly divided between primary and secondary porosity. Primary porosity was preserved by early, iron-rich grain-rimming chlorite and quartz overgrowths. Most secondary porosity resulted from dissolution of carbonate cements. Moldic pores outlined by chlorite were created by dissolution of unstable feldspars and rock fragments. Interparticle clay microporosity is significant in sandstones containing authigenic kaolinite and (or) chlorite. Pores were filled in the deepest sandstones by quartz overgrowths and a late magnesium-rich chlorite that is commonly obscured by fibrous illite. Voids were created in the early Tertiary(?) by acidic meteoric waters and during deep burial by brines carrying organic and inorganic acids that were released during hydrocarbon maturation in neighboring shales. Oil fills dissolution voids in ankerite cement and albitized plagioclase and coats most authigenic minerals. Two-phase primary fluid inclusions in quartz overgrowths which also contain oil-bearing inclusions have homogenization temperatures between 125{degrees}C and 134{degrees}C. These temperatures combined with a burial history reconstruction indicate that hydrocarbons migrated into Tuscaloosa sandstones during the Miocene. Overpressuring began in the middle Tertiary along with gas generation in the Tuscaloosa. These events coincided with the end of deep meteoric flow through the Gulf section and the beginning of a compactional hydrologic regime. Precipitation of quartz overgrowths and hydrocarbons at this time locally created effective pressure seals.

  3. Planet formation and searches

    NASA Astrophysics Data System (ADS)

    Montgomery, Ryan Michael

    2009-08-01

    This thesis explores the possibilities for discovery of terrestrial-mass planets in the habitable zones of their host stars. Towards this aim, we present the results of three projects and discuss another two preliminary studies of further explorations. In so doing, we explore a fairly comprehensive range of possibilities regarding the formation and detection of terrestrial- mass planets in the habitable zone. We first study the potential for terrestrial planets to form in situ in and around the habitable zones of M-dwarf stars. We proceed to explore the feasibility of searches for these planets using the transit method via Monte- Carlo simulations. We find that M-dwarfs pose an interesting challenge for study: being inherently dim, widely spread on the sky, and photometrically variable. We present results of simulated ground-based transit search campaigns as well as simulated searches from a modest satellite mission. Our second project is a straightforward extension of the previous study: a collaborative effort to search for transit signals around the nearest M-dwarf: Proxima Centauri. We describe our observations as well as the Monte-Carlo analysis used to place constraints on the possible planetary radii and periods. Our third project is a search for transiting extra-solar Jovian planets using the Rossiter-McLaughlin effect. We search through the private Keck radial- velocity datasets for undiscovered Rossiter-McLaughlin signals. We present our results in the form of both strong null-result datasets as well as potential transiting systems. We then briefly analyze these larger Jovian planets for potential to harbor potentially habitable terrestrial satellites. Our final preliminary analysis looks into the potential for the Large Synoptic Survey Telescope to detect transiting Neptune-mass planets orbiting M-dwarfs which could then lead to terrestrial-mass planet detections. The sum of these efforts is a comprehensive investigation into the likelihood and

  4. Medusae Fossae Formation

    NASA Technical Reports Server (NTRS)

    1998-01-01

    An exotic terrain of wind-eroded ridges and residual smooth surfaces are seen in one of the highest resolution images ever taken of Mars from orbit. The Medusae Fossae formation is believed to be formed of the fragmental ejecta of huge explosive volcanic eruptions. When subjected to intense wind-blasting over hundreds of millions of years, this material erodes easily once the uppermost tougher crust is breached. In the Mars Orbiter Camera (MOC) image shown on the right, the crust, or cap rock, can be seen in the upper right part of the picture. The finely-spaced ridges are similar to features on Earth called yardangs, which are formed by intense winds plucking individual grains from, and by wind-driven sand blasting particles off, sedimentary deposits.

    The MOC image was taken on October 30, 1997 at 11:05 AM PST, shortly after the Mars Global Surveyor spacecraft's 31st closest approach to Mars. The image covers an area 3.6 X 21.5 km (2.2 X 13.4 miles) at 3.6 m (12 feet) per picture element--craters only 11 m (36 feet, about the size of a swimming pool) across can be seen. The context image (left; the best Viking view of the area; VO 1 387S34) has a resolution of 240 m/pixel, or 67 times lower resolution than the MOC frame.

    Malin Space Science Systems (MSSS) and the California Institute of Technology built the MOC using spare hardware from the Mars Observer mission. MSSS operates the camera from its facilities in San Diego, CA. The Jet Propulsion Laboratory's Mars Surveyor Operations Project operates the Mars Global Surveyor spacecraft with its industrial partner, Lockheed Martin Astronautics, from facilities in Pasadena, CA and Denver, CO.

  5. Formative Assessment Probes: Is It a Rock? Continuous Formative Assessment

    ERIC Educational Resources Information Center

    Keeley, Page

    2013-01-01

    A lesson plan is provided for a formative assessment probe entitled "Is It a Rock?" This probe is designed for teaching elementary school students about rocks through the use of a formative assessment classroom technique (FACT) known as the group Frayer Model. FACT activates students' thinking about a concept and can be used to…

  6. A high-throughput assay format for determination of nitrate reductase and nitrite reductase enzyme activities

    SciTech Connect

    McNally, N.; Liu, Xiang Yang; Choudary, P.V.

    1997-01-01

    The authors describe a microplate-based high-throughput procedure for rapid assay of the enzyme activities of nitrate reductase and nitrite reductase, using extremely small volumes of reagents. The new procedure offers the advantages of rapidity, small sample size-nanoliter volumes, low cost, and a dramatic increase in the throughput sample number that can be analyzed simultaneously. Additional advantages can be accessed by using microplate reader application software packages that permit assigning a group type to the wells, recording of the data on exportable data files and exercising the option of using the kinetic or endpoint reading modes. The assay can also be used independently for detecting nitrite residues/contamination in environmental/food samples. 10 refs., 2 figs.

  7. The Formation of Galactic Bulges

    NASA Astrophysics Data System (ADS)

    Carollo, C. Marcella; Ferguson, Henry C.; Wyse, Rosemary F. G.

    2000-03-01

    Part I. Introduction: What are galactic bulges?; Part II. The Epoch of Bulge Formation: Origin of bulges; Deep sub-mm surveys: High-z ULIRGs and the formation of spheroids; Ages and metallicities for stars in the galactic bulge; Integrated stellar populations of bulges: First results; HST-NICMOS observations of galactic bulges: Ages and dust; Inside-out bulge formation and the origin of the Hubble sequence; Part III. The Timescales of Bulge Formation: Constraints on the bulge formation timescale from stellar populations; Bulge building with mergers and winds; Role of winds, starbursts, and activity in bulge formation; Dynamical timescales of bulge formation; Part IV. Physical Processes in Bulge Formation: the role of bars for secular bulge formation; Bars and boxy/peanut-shaped bulges: an observational point of view; Boxy- and peanut-shaped bulges; A new class of bulges; The role of secondary bars in bulge formation; Radial transport of molecular gas to the nuclei of spiral galaxies; Dynamical evolution of bulge shapes; Two-component stellar systems: Phase-space constraints; Central NGC 2146 - a firehose-type bending instability?; Bulge formation: the role of the multi-phase ISM; Global evolution of a self-gravitating multi-phase ISM in the central kpc region of galaxies; Part V. Bulge Phenomenology: Bulge-disk decomposition of spiral galaxies in the near-infrared; The triaxial bulge of NGC 1371; The bulge-disk orthogonal decoupling in galaxies: NGC 4698 and NGC 4672; The kinematics and the origin of the ionized gas in NGC 4036; Optically thin thermal plasma in the galactic bulge; X-ray properties of bulges; The host galaxies of radio-loud AGN; The centers of radio-loud early-type galaxies with HST; Central UV spikes in two galactic spheroids; Conference summary: where do we stand?

  8. The formation of Pangea

    NASA Astrophysics Data System (ADS)

    Stampfli, G. M.; Hochard, C.; Vérard, C.; Wilhem, C.; vonRaumer, J.

    2013-05-01

    The making of Pangea is the result of large-scale amalgamation of continents and micro-continents, which started at the end of the Neoproterozoic with the formation of Gondwana. As pieces were added to Gondwana on its South-American, Antarctica and Australia side, ribbon-like micro-continents were detached from its African and South-Chinese side: Cadomia in the late Neoproterozoic, Avalonia and Hunia in the Ordovician, Galatia in the Devonian and Cimmeria in the Permian. Cadomia was re-accreted to Gondwana, but the other ribbon-continents were accreted to Baltica, North-China, Laurussia or Laurasia. Finding the origin of these numerous terranes is a major geological challenge. Recently, a global plate tectonic model was developed together with a large geological/geodynamic database, at the Lausanne University, covering the last 600 Ma of the Earth's history. Special attention was given to the placing of Gondwana derived terranes in their original position, using all possible constraints. We propose here a solution for the Variscan terranes, another paper deals with the Altaids. The Galatian super-terrane was detached from Gondwana in the Devonian, during the opening of Paleotethys, and was quickly separated into four sub-terranes that started to by-pass each other. The leading terranes collided at the end of the Devonian with the Hanseatic terrane detached from Laurussia. In the Carboniferous, Gondwana started to impinge onto the amalgamated terranes, creating the Variscan chain and the Pangean super-continent. East of Spain Paleotethys remained opened until the Triassic, subducting northward under Laurasia. Roll-back of the Paleotethyan slab triggered the collapse of most of the European Variscan orogen, which was replaced by series of Permian rifts, some of them becoming oceanized back-arc basins during the Triassic. Major force changes at the Pangean plate limits at the end of the Triassic provoked its break-up, through the opening of the proto

  9. Formation Algorithms and Simulation Testbed

    NASA Technical Reports Server (NTRS)

    Wette, Matthew; Sohl, Garett; Scharf, Daniel; Benowitz, Edward

    2004-01-01

    Formation flying for spacecraft is a rapidly developing field that will enable a new era of space science. For one of its missions, the Terrestrial Planet Finder (TPF) project has selected a formation flying interferometer design to detect earth-like planets orbiting distant stars. In order to advance technology needed for the TPF formation flying interferometer, the TPF project has been developing a distributed real-time testbed to demonstrate end-to-end operation of formation flying with TPF-like functionality and precision. This is the Formation Algorithms and Simulation Testbed (FAST) . This FAST was conceived to bring out issues in timing, data fusion, inter-spacecraft communication, inter-spacecraft sensing and system-wide formation robustness. In this paper we describe the FAST and show results from a two-spacecraft formation scenario. The two-spacecraft simulation is the first time that precision end-to-end formation flying operation has been demonstrated in a distributed real-time simulation environment.

  10. Formative Assessment: A Critical Review

    ERIC Educational Resources Information Center

    Bennett, Randy Elliot

    2011-01-01

    This paper covers six interrelated issues in formative assessment (aka, "assessment for learning"). The issues concern the definition of formative assessment, the claims commonly made for its effectiveness, the limited attention given to domain considerations in its conceptualisation, the under-representation of measurement principles in that…

  11. New Frontiers in Formative Assessment

    ERIC Educational Resources Information Center

    Noyce, Pendred E., Ed.; Hickey, Daniel T., Ed.

    2011-01-01

    "Formative assessment is a powerful learning tool that is too seldom, too haphazardly, and too ineffectively used in the United States," Pendred E. Noyce writes in the introduction to this volume. "The purpose of this book is to delve into why this is so and how it can be changed." Formative assessment involves constantly monitoring student…

  12. The Principal as Formative Coach

    ERIC Educational Resources Information Center

    Nidus, Gabrielle; Sadder, Maya

    2011-01-01

    Formative coaching, an approach that uses student work as the foundation for mentoring and professional development, can help principals become more effective instructional leaders. In formative coaching, teaches and coaches analyze student work to determine next steps for instruction. This article shows how a principal can use the steps of the…

  13. FORMATION OF SECONDARY ORGANIC AEROSOL

    EPA Science Inventory

    (1) Gas-phase chemistry. With the clear and profound effect of the VOC/NOx ratio on SOA formation, we will augment gas-phase VOC oxidation mechanisms in atmospheric models to account for the effect of NOx level on the mechanism of SOA formation; (2) Revis...

  14. Portable File Format (PFF) specifications.

    SciTech Connect

    Dolan, Daniel H.,

    2015-02-01

    Created at Sandia National Laboratories, the Portable File Format (PFF) allows binary data transfer across computer platforms. Although this capability is supported by many other formats, PFF files are still in use at Sandia, particularly in pulsed power research. This report provides detailed PFF specifications for accessing data without relying on legacy code.

  15. Star formation in the multiverse

    SciTech Connect

    Bousso, Raphael; Leichenauer, Stefan

    2009-03-15

    We develop a simple semianalytic model of the star formation rate as a function of time. We estimate the star formation rate for a wide range of values of the cosmological constant, spatial curvature, and primordial density contrast. Our model can predict such parameters in the multiverse, if the underlying theory landscape and the cosmological measure are known.

  16. The Apennine Bench Formation revisited

    NASA Technical Reports Server (NTRS)

    Spudis, P. D.; Hawke, B. R.

    1985-01-01

    The Apennine Bench Formation consists of pre-mare light plains materials that crop out south of the crater Archimedes, inside the Imbrium basin. This material was ascribed to either impact or volcanic origins. The characteristics of Apollo 15 KREEP basalts and the Apennine Bench Formation are reviewed to determine whether their characteristics are compatible with a volcanic origin.

  17. SAS FORMATS: USES AND ABUSES

    EPA Science Inventory

    SAS formats are a very powerful tool. They allow you to display the data in a more readable manner without modifying it. Formats can also be used to group data into categories for use in various procedures like PROC FREQ, PROC TTEST, and PROC MEANS (as a class variable). As w...

  18. SAS FORMATS: USES AND ABUSES

    EPA Science Inventory

    SAS formats are a very powerful tool. They allow you to display the data in a more readable manner without modifying it. Formats can also be used to group data into categories for use in various procedures like PROC FREQ, PROC TTEST, and PROC MEANS (as a class variable). As ...

  19. Physics of primordial star formation

    NASA Astrophysics Data System (ADS)

    Yoshida, Naoki

    2012-09-01

    The study of primordial star formation has a history of nearly sixty years. It is generally thought that primordial stars are one of the key elements in a broad range of topics in astronomy and cosmology, from Galactic chemical evolution to the formation of super-massive blackholes. We review recent progress in the theory of primordial star formation. The standard theory of cosmic structure formation posits that the present-day rich structure of the Universe developed through gravitational amplification of tiny matter density fluctuations left over from the Big Bang. It has become possible to study primordial star formation rigorously within the framework of the standard cosmological model. We first lay out the key physical processes in a primordial gas. Then, we introduce recent developments in computer simulations. Finally, we discuss prospects for future observations of the first generation of stars.

  20. Core formation by giant impacts

    NASA Technical Reports Server (NTRS)

    Tonks, W. B.; Melosh, H. J.

    1991-01-01

    Ideas about the accretion and early evolution of the Earth and the other terrestrial planets have recently undergone a number of revolutionary changes. It has become clear that giant impacts were far from rare events. In the later stages of accretion any given planetary embryo is liable to be struck several times by other bodies of up to half its own diameter. Such an impact may have the ability to trigger core formation. Traditional accretion models have had great difficulty explaining the formation of the core. If one admits the importance of infrequent large events that may melt an entire hemisphere, the core formation difficulty vanishes. Millimeter-size iron blebs in the melted region will rain out due to their density difference with the silicate melt. Core formation may not require the melting of the entire hemisphere of the planet. The conditions are explored under which impact induced core formation may occur.

  1. Development of resazurin-based assay in 384-well format for high throughput whole cell screening of Trypanosoma brucei rhodesiense strain STIB 900 for the identification of potential anti-trypanosomal agents.

    PubMed

    Lim, Kah Tee; Zahari, Zuriati; Amanah, Azimah; Zainuddin, Zafarina; Adenan, Mohd Ilham

    2016-03-01

    To accelerate the discovery of novel leads for the treatment of Human African Trypanosomiasis (HAT), it is necessary to have a simple, robust and cost-effective assay to identify positive hits by high throughput whole cell screening. Most of the fluorescence assay was made in black plate however in this study the HTS assay developed in 384-well format using clear plate and black plate, for comparison. The HTS assay developed is simple, sensitive, reliable and reproducible in both types of plates. Assay robustness and reproducibility were determined under the optimized conditions in 384-well plate was well tolerated in the HTS assay, including percentage of coefficient of variation (% CV) of 4.68% and 4.74% in clear and black 384-well plate, signal-to-background ratio (S/B) of 12.75 in clear 384-well plate and 12.07 in black 384-well plate, Z' factor of 0.79 and 0.82 in clear 384-well plate and black 384-well plate, respectively and final concentration of 0.30% dimethylsulfoxide (DMSO) in both types of plate. Drug sensitivity was found to be comparable to the reported anti-trypanosomal assay in 96-well format. The reproducibility and sensitivity of this assay make it compliant to automated liquid handler use in HTS applications. PMID:26772786

  2. Dynamics of Earth orbiting formations

    NASA Technical Reports Server (NTRS)

    Ploen, Scott R.; Scharf, Daniel P.; Hadaegh, Fred Y.; Acikmese, Ahmed B.

    2004-01-01

    In this paper the equations of motion of a formation consisting of n spacecraft in Earth orbit are derived via Lagrange's equations. The equations of motion of the formation are developed with respect to both (1) a bound Keplerian reference orbit, and (2) a specific spacecraft in the formation. The major orbital perturbations acting on a formation in low Earth orbit are also included in the analysis. In contrast to the traditional approach based on the balance of linear momentum, the use of Lagrange's equations leads to a high-level matrix derivation of the formation equations of motion. The matrix form of the nonlinear motion equations is then linearized about a bound Keplerian reference orbit. Next, it is demonstrated that under the assumption of a circular reference orbit, the linearized equations of motion reduce to the well-known Hill-Clohessy-Wiltshire equations. The resulting linear and nonlinear dynamic equations lead to maximal physical insight into the structure of formation dynamics, and are ideally suited for use in the design and validation of formation guidance and control laws.

  3. Dynamics of rock varnish formation

    SciTech Connect

    Raymond, R. Jr.; Reneau, S.L.; Guthrie, G.D. Jr.; Bish, D.L.; Harrington, C.D.

    1991-01-01

    Our studies of rock varnish from the southwestern United States suggest that the Mn-phase in rock varnish has neither the chemistry nor the crystal structure of birnessite. Rather, the Mn-rich phase is non-crystalline and contains Ba, Ca, Fe, Al, and P. Unknowns concerning the formation of this non-crystalline Mn phase must be resolved before researchers are able to define chemical parameters of rock varnish formation based upon conditions of formation of the Mn phase. 6 refs., 9 figs.

  4. Antihydrogen Formation using Cold Plasmas

    SciTech Connect

    Madsen, N.; Bowe, P.D.; Hangst, J.S.; Amoretti, M.; Carraro, C.; Macri, M.; Testera, G.; Variola, A.; Amsler, C.; Johnson, I.; Pruys, H.; Regenfus, C.; Bonomi, G.; Bouchta, A.; Doser, M.; Kellerbauer, A.; Landua, R.; Cesar, C.L.; Charlton, M.; Joergensen, L.V.

    2004-10-20

    Antihydrogen, the antimatter counterpart of the hydrogen atom, can be formed by mixing cold samples of antiprotons and positrons. In 2002 the ATHENA collaboration succeeded in the first production of cold antihydrogen. By observing and imaging the annihilation products of the neutral, non-confined, antihydrogen atoms annihilating on the walls of the trap we can observe the production in quasi-real-time and study the dynamics of the formation mechanism. The formation mechanism strongly influences the final state of the formed antihydrogen atoms, important for future spectroscopic comparison with hydrogen. This paper briefly summarizes the current understanding of the antihydrogen formation in ATHENA.

  5. Dissipative processes in galaxy formation.

    PubMed Central

    Silk, J

    1993-01-01

    A galaxy commences its life in a diffuse gas cloud that evolves into a predominantly stellar aggregation. Considerable dissipation of gravitational binding energy occurs during this transition. I review here the dissipative processes that determine the critical scales of luminous galaxies and the generation of their morphology. The universal scaling relations for spirals and ellipticals are shown to be sensitive to the history of star formation. Semiphenomenological expressions are given for star-formation rates in protogalaxies and in starbursts. Implications are described for elliptical galaxy formation and for the evolution of disk galaxies. PMID:11607396

  6. Dynamics of interfacial pattern formation

    NASA Technical Reports Server (NTRS)

    Ben-Jacob, E.; Goldenfeld, N.; Langer, J. S.; Schon, G.

    1983-01-01

    A phenomenological model of dendritic solidification incorporating interfacial kinetics, crystalline anisotropy, and a local approximation for the dynamics of the thermal diffusion field is proposed. The preliminary results are in qualitative agreement with natural dendrite-like pattern formation.

  7. Conduction heating of hydrocarbonaceous formations

    SciTech Connect

    Bridges, J. E.

    1985-10-08

    A waveguide structure is emplanted in the earth to bound a particular volume of an earth formation with a waveguide structure formed of respective rows of discrete elongated electrodes wherein the spacing between rows is greater than the distance between electrodes in a respective row and in the case of vertical electrodes substantially less than the thickness of the hydrocarbonaceous earth formation. Electrical power at no more than a relatively low frequency is applied between respective rows of the electrodes to deliver power to the formation while producing relatively uniform heating thereof and limiting the relative loss of heat to adjacent barren regions to less than a tolerable amount. At the same time the temperature of the electrodes is controlled near the vaporization point of water thereat to maintain an electrically conductive path between the electrodes and the formation.

  8. Maximum likelihood topographic map formation.

    PubMed

    Van Hulle, Marc M

    2005-03-01

    We introduce a new unsupervised learning algorithm for kernel-based topographic map formation of heteroscedastic gaussian mixtures that allows for a unified account of distortion error (vector quantization), log-likelihood, and Kullback-Leibler divergence. PMID:15802004

  9. Immigration, Integration and Ghetto Formation

    NASA Astrophysics Data System (ADS)

    Meyer-Ortmanns, Hildegard

    We study ghetto formation in a population with natives and immigrants in the framework of the two-dimensional Ising-model with Kawasaki-exchange dynamics. It is the phase structure of the Ising model, the integration speed and the immigration rate which determine whether ghetto formation between natives and immigrants can be avoided or not. Our simulations are performed in- and out-of-equilibrium.

  10. Cosmic strings and galaxy formation

    NASA Technical Reports Server (NTRS)

    Bertschinger, Edmund

    1989-01-01

    The cosmogonical model proposed by Zel'dovich and Vilenkin (1981), in which superconducting cosmic strings act as seeds for the origin of structure in the universe, is discussed, summarizing the results of recent theoretical investigations. Consideration is given to the formation of cosmic strings, the microscopic structure of strings, gravitational effects, cosmic string evolution, and the formation of galaxies and large-scale structure. Simulation results are presented in graphs, and several outstanding issues are listed and briefly characterized.

  11. Modes of clustered star formation

    NASA Astrophysics Data System (ADS)

    Pfalzner, S.; Kaczmarek, T.; Olczak, C.

    2012-09-01

    Context. The recent realization that most stars form in clusters, immediately raises the question of whether star and planet formation are influenced by the cluster environment. The stellar density in the most prevalent clusters is the key factor here. Whether dominant modes of clustered star formation exist is a fundamental question. Using near-neighbour searches in young clusters, Bressert and collaborators claim this not to be the case. They conclude that - at least in the solar neighbourhood - star formation is continuous from isolated to densely clustered environments and that the environment plays a minor role in star and planet formation. Aims: We investigate under which conditions near-neighbour searches in young clusters can distinguish between different modes of clustered star formation. Methods: Model star clusters with different memberships and density distributions are set up and near-neighbour searches are performed. We investigate the influence of the combination of different cluster modes, observational biases, and types of diagnostic on the results. Results: We find that the specific cluster density profile, the relative sample sizes, the limitations of the observation, and the choice of diagnostic method decide, whether modelled modes of clustered star formation are detected by near-neighbour searches. For density distributions that are centrally concentrated but span a wide density range (for example, King profiles), separate cluster modes are only detectable under ideal conditions (sample selection, completeness) if the mean density of the individual clusters differs by at least a factor of ~65. Introducing a central cut-off can lead to an underestimate of the mean density by more than a factor of ten especially in high density regions. The environmental effect on star and planet formation is similarly underestimated for half of the population in dense systems. Conclusions: Local surface-density distributions are a very useful tool for single

  12. Treating tar sands formations with karsted zones

    SciTech Connect

    Vinegar, Harold J.; Karanikas, John Michael

    2010-03-09

    Methods for treating a tar sands formation are described herein. The tar sands formation may have one or more karsted zones. Methods may include providing heat from one or more heaters to one or more karsted zones of the tar sands formation to mobilize fluids in the formation. At least some of the mobilized fluids may be produced from the formation.

  13. The Dynamics of Latifundia Formation

    PubMed Central

    Chaves, Luis Fernando

    2013-01-01

    Land tenure inequity is a major social problem in developing nations worldwide. In societies, where land is a commodity, inequities in land tenure are associated with gaps in income distribution, poverty and biodiversity loss. A common pattern of land tenure inequities through the history of civilization has been the formation of latifundia [Zhuāngyuán in chinese], i.e., a pattern where land ownership is concentrated by a small fraction of the whole population. Here, we use simple Markov chain models to study the dynamics of latifundia formation in a heterogeneous landscape where land can transition between forest, agriculture and recovering land. We systematically study the likelihood of latifundia formation under the assumption of pre-capitalist trade, where trade is based on the average utility of land parcels belonging to each individual landowner during a discrete time step. By restricting land trade to that under recovery, we found the likelihood of latifundia formation to increase with the size of the system, i.e., the amount of land and individuals in the society. We found that an increase of the transition rate for land use changes, i.e., how quickly land use changes, promotes more equitable patterns of land ownership. Disease introduction in the system, which reduced land profitability for infected individual landowners, promoted the formation of latifundia, with an increased likelihood for latifundia formation when there were heterogeneities in the susceptibility to infection. Finally, our model suggests that land ownership reforms need to guarantee an equitative distribution of land among individuals in a society to avoid the formation of latifundia. PMID:24376597

  14. The dynamics of latifundia formation.

    PubMed

    Chaves, Luis Fernando

    2013-01-01

    Land tenure inequity is a major social problem in developing nations worldwide. In societies, where land is a commodity, inequities in land tenure are associated with gaps in income distribution, poverty and biodiversity loss. A common pattern of land tenure inequities through the history of civilization has been the formation of latifundia [Zhuāngyuán in chinese], i.e., a pattern where land ownership is concentrated by a small fraction of the whole population. Here, we use simple Markov chain models to study the dynamics of latifundia formation in a heterogeneous landscape where land can transition between forest, agriculture and recovering land. We systematically study the likelihood of latifundia formation under the assumption of pre-capitalist trade, where trade is based on the average utility of land parcels belonging to each individual landowner during a discrete time step. By restricting land trade to that under recovery, we found the likelihood of latifundia formation to increase with the size of the system, i.e., the amount of land and individuals in the society. We found that an increase of the transition rate for land use changes, i.e., how quickly land use changes, promotes more equitable patterns of land ownership. Disease introduction in the system, which reduced land profitability for infected individual landowners, promoted the formation of latifundia, with an increased likelihood for latifundia formation when there were heterogeneities in the susceptibility to infection. Finally, our model suggests that land ownership reforms need to guarantee an equitative distribution of land among individuals in a society to avoid the formation of latifundia. PMID:24376597

  15. Modeling of Mitochondrial Donut Formation.

    PubMed

    Long, Qi; Zhao, Danyun; Fan, Weimin; Yang, Liang; Zhou, Yanshuang; Qi, Juntao; Wang, Xin; Liu, Xingguo

    2015-09-01

    Mitochondria are highly dynamic cell organelles. Continual cycles of fusion and fission play an important role in mitochondrial metabolism and cellular signaling. Previously, a novel mitochondrial morphology, the donut, was reported in cells after hypoxia-reoxygenation or osmotic pressure changes. However, the mechanism of donut formation remained elusive. Here, we obtained the distribution of donut diameters (D = 2R) and found that 95% are >0.8 μm. We also performed highly precise measurements of the mitochondrial tubule diameters using superresolution and electron microscopy. Then, we set up a model by calculating the mitochondrial bending energy and osmotic potential during donut formation. It shows that the bending energy is increased as the radius of curvature, R, gets smaller in the process of donut formation, especially for radii <0.4 μm, creating a barrier to donut formation. The calculations also show that osmotic potential energy release can balance the rising bending energy through volume expansion. Finally, we revealed the donut formation process in a Gibbs free-energy-dependent model combining calculations and measurements. PMID:26331247

  16. The Physics of Planetesimal Formation

    NASA Astrophysics Data System (ADS)

    Simon, Jacob; Armitage, Philip; Youdin, Andrew; Li, Rixin

    2015-12-01

    Planetesimals are the precursors to planets, and understanding their formation is an essential step towards developing a complete theory of planet formation. For small solid particles (e.g., dust grains) to coagulate into planetesimals, however, requires that these particles grow beyond centimeter sizes; with traditional coagulation physics, this is very difficult. The streaming instability, which is a clumping process akin to the pile-up of cars in a traffic jam, generates sufficiently high solid densities that the mutual gravity between the clumped particles eventually causes their collapse towards planetesimal mass and size scales. Exploring this transition from dust grains to planetesimals is still in its infancy but is extremely important if we want to understand the basics of planet formation. Here, I present a series of high resolution, first principles numerical simulations of potoplanetary disk gas and dust to study the clumping of particles via the streaming instability and the subsequent collapse towards planetesimals. These simulations have been employed to characterize the planetesimal population as a function of radius in protoplanetary disks. The results of these simulations will be crucial for planet formation models to correctly explain the formation and configuration of solar systems.

  17. Star formation in Galactic flows

    NASA Astrophysics Data System (ADS)

    Smilgys, Romas; Bonnell, Ian A.

    2016-06-01

    We investigate the triggering of star formation in clouds that form in Galactic scale flows as the interstellar medium passes through spiral shocks. We use the Lagrangian nature of smoothed particle hydrodynamics simulations to trace how the star-forming gas is gathered into self-gravitating cores that collapse to form stars. Large-scale flows that arise due to Galactic dynamics create shocks of the order of 30 km s-1 that compress the gas and form dense clouds (n > several × 102 cm-3) in which self-gravity becomes relevant. These large-scale flows are necessary for creating the dense physical conditions for gravitational collapse and star formation. Local gravitational collapse requires densities in excess of n > 103 cm-3 which occur on size scales of ≈1 pc for low-mass star-forming regions (M < 100 M⊙), and up to sizes approaching 10 pc for higher mass regions (M > 103 M⊙). Star formation in the 250 pc region lasts throughout the 5 Myr time-scale of the simulation with a star formation rate of ≈10-1 M⊙ yr-1 kpc-2. In the absence of feedback, the efficiency of the star formation per free-fall time varies from our assumed 100 per cent at our sink accretion radius to values of <10-3 at low densities.

  18. SW New Mexico Oil Well Formation Tops

    DOE Data Explorer

    Shari Kelley

    2015-10-21

    Rock formation top picks from oil wells from southwestern New Mexico from scout cards and other sources. There are differing formation tops interpretations for some wells, so for those wells duplicate formation top data are presented in this file.

  19. Formative Evaluation in the Performance Context.

    ERIC Educational Resources Information Center

    Dick, Walter; King, Debby

    1994-01-01

    Reviews the traditional formative evaluation model used by instructional designers; summarizes Kirkpatrick's model of evaluation; proposes the integration of part of Kirkpatrick's model with traditional formative evaluation; and discusses performance-context formative evaluation. (three references) (LRW)

  20. Fluorimetric screening assay for protein carbonyl evaluation in biological samples.

    PubMed

    Stocker, P; Ricquebourg, E; Vidal, N; Villard, C; Lafitte, D; Sellami, L; Pietri, S

    2015-08-01

    Many assays are available for the detection of protein carbonyls (PCs). Currently, the measurement of PC groups after their derivatization with 2,4-dinitrophenol hydrazine (DNPH) is widely used for measuring protein oxidation in biological samples. However, this method includes several washing steps. In this context, we have developed a rapid, sensitive, and accurate fluorimetric method adapted to 96-well microplates for the convenient assessment of protein carbonyl level in biological samples. The method reported here is based on the reaction of carbonyl content in proteins with 7-hydrazino-4-nitrobenzo-2,1,3-oxadiazole (NBDH) to form highly fluorescent derivatives via hydrazone formation. PCs were determined using the DNPH and NBDH assays in fully reduced bovine serum albumin (BSA) and plasma and liver homogenates obtained from healthy control rats up the addition of various amounts of HOCl-oxidized BSA (OxBSA). Using the NBDH assay, PC concentrations as low as 0.2 nmol/mg were detected with precision as low as 5%. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectroscopy was used to successfully identify the formation of the NBDH adducts after derivatization with standard oxidized peptides. Finally, the two methods were further used for PC determination in plasma and liver samples from diabetic and normal rats, showing that the NBDH assay can be reliably used in biological experiments. PMID:25933703