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Sample records for 96-well microplate format

  1. A highly sensitive and versatile virus titration assay in the 96-well microplate format.

    PubMed

    Borisevich, V; Nistler, R; Hudman, D; Yamshchikov, G; Seregin, A; Yamshchikov, V

    2008-02-01

    This report describes a fast, reproducible, inexpensive and convenient assay system for virus titration in the 96-well format. The micromethod substantially increases assay throughput and improves the data reproducibility. A highly simplified variant of virus quantification is based on immunohistochemical detection of virus amplification foci obtained without use of agarose or semisolid overlays. It can be incorporated into several types of routine virological assays successfully replacing the laborious and time-consuming conventional methods based on plaque formation under semisolid overlays. The method does not depend on the development of CPE and can be accommodated to assay viruses with substantial differences in growth properties. The use of enhanced immunohistochemical detection enabled a five- to six-fold reduction of the total assay time. The micromethod was specifically developed to take advantage of multichannel pipettor use to simplify handling of a large number of samples. The method performs well with an inexpensive low-power binocular, thus offering a routine assay system usable outside of specialized laboratory setting, such as for testing of clinical or field samples. When used in focus reduction-neutralization tests (FRNT), the method accommodates very small volumes of immune serum, which is often a decisive factor in experiments involving small rodent models.

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

    PubMed

    Bi, Xiaodong; Liu, Zhen

    2014-12-16

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

  3. Detection of enteroviruses in shellfish by fluorogenic polymerase chain reaction integrated with 96-well microplate scanning.

    PubMed

    Shieh, Y Carol; Baric, Ralph S

    2002-01-01

    A one-step procedure was developed to confirm viral targets by using a fluorometric 96-well microplate scanner following polymerase chain reaction (PCR). The fluorogenic PCR, integrated with fluorometric scanning, measured the end point fluorescence of viral PCR amplicon/probe hybrids and permitted the use of nonfluorogenic PCR conditions with addition of a Cy3 fluorophore-labeled linear probe for viruses. This linear probe generated higher ratios of viral signal-to-noise than a comparative beacon probe. Detection efficiency with a Cy3/quencher linear probe was comparable with Southern analysis at the level > or = 0.27 plaque-forming units (PFU) of poliovirus/PCR. For the reaction containing < 0.27 PFU, the fluorometric measurements of the first-round PCR viral amplicon were not as sensitive as Southern analysis; however, equivalent sensitivities were achieved with fluorogenic nested PCR. Concentrates of 11 oyster samples exposed to municipal sewage were tested for enteroviruses; the fluorogenic detection correlated 100% with Southern analysis. This method using fluorometric scanning of viral amplicon is simple; it requires neither continuously monitoring equipment nor redesigning PCR primers; and it accurately detects enteroviruses in oyster sample concentrates in less time than classic spectrophotometry or Southern analysis.

  4. 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.

  5. 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}.

  6. Facile preparation of glycoprotein-imprinted 96-well microplates for enzyme-linked immunosorbent assay by boronate affinity-based oriented surface imprinting.

    PubMed

    Bi, Xiaodong; Liu, Zhen

    2014-01-01

    Molecularly imprinted polymers (MIPs), as inexpensive and stable substitutes of antibodies, have shown great promise in immunoassays. Glycoproteins are of significant diagnostic value. To facilitate the application of MIPs in clinical diagnostics, a general and facile imprinting method toward glycoproteins oriented for an enzyme-linked immunosorbent assay (ELISA) in the form of a 96-well microplate is essential but has not been fully explored yet. In this study, a new method called boronate affinity-based oriented surface imprinting was proposed for facile preparation of glycoprotein-imprinted microplates. A template glycoprotein was first immobilized by a boronic acid-modified microplate through boronate affinity binding, and then, a thin layer of polyaniline was formed to cover the microplate surface via in-water self-copolymerization. After the template was removed by an acidic solution, 3D cavities that can rebind the template were fabricated on the microplate surface. Using horseradish peroxidase (HRP) as a model target, the effects of imprinting conditions as well as the properties and performance of the prepared MIPs were investigated. α-Fetoprotein (AFP)-imprinted microplate was then prepared, and thereby, a MIP-based ELISA method was established. The prepared MIPs exhibited several highly favorable features, including excellent specificity, widely applicable binding pH, superb tolerance for interference, high binding strength, fast equilibrium kinetics, and reusability. The MIP-based ELISA method was finally applied to the analysis of AFP in human serum. The result was in good agreement with that by radioimmunoassay, showing a promising prospect of the proposed method in clinical diagnostics.

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

    EPA Science Inventory

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

  8. GT1-7 cell-based cytoxicity screening assay on 96-well microplates as a platform for the safety assessment of genetically modified Gerbera hybrida extracts.

    PubMed

    Fallarero, Adyary; Ainasoja, Miia; Sandberg, Malena; Teeri, Teemu H; Vuorela, Pia M

    2009-01-01

    In this investigation, a GT1-7 cell-based cytotoxicity screening assay in 96-well microplates was set up. The assay, using propidium iodide fluorescence, was proven to be reliable, with good quality (Z' = 0.51) and low plate-to-plate and day-to-day variations. Further on, a library containing extracts from 227 genetic modification (GM) Gerbera hybrida and 42 Gerbera varieties was screened; however, no differences between them were found. Based on these findings, we propose the use of the current assay within the first-tier screening studies of large collections. Also, these results provide valuable information for GM Gerbera risk-assessment purposes and offer a model for the toxicity cell-based screening of GM crops.

  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.

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

    PubMed Central

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

    2016-01-01

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

  11. Automated expression and solubility screening of His-tagged proteins in 96-well format.

    PubMed

    Vincentelli, Renaud; Canaan, Stéphane; Offant, Julien; Cambillau, Christian; Bignon, Christophe

    2005-11-01

    A growing need for sensitive and high-throughput methods for screening the expression and solubility of recombinant proteins exists in structural genomics. Originally, the emergency solution was to use immediately available techniques such as manual lysis of expression cells followed by analysis of protein expression by gel electrophoresis. However, these handmade methods quickly proved to be unfit for the high-throughput demand of postgenomics, and it is now generally accepted that the long-term solution to this problem will be based on automation, on industrial standard-formatted experiments, and on downsizing samples and consumables. In agreement with this consensus, we have set up a fully automated method based on a dot-blot technology and using 96-well format consumables for assessing by immunodetection the amount of total and soluble recombinant histidine (His)-tagged proteins expressed in Escherichia coli. The method starts with the harvest of expression cells and ends with the display of solubility/expression results in milligrams of recombinant protein per liter of culture using a three-color code to assist analysis. The program autonomously processes 160 independent cultures at a time.

  12. 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

  13. 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

  14. 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.

  15. 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

  16. 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

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

    PubMed

    Cudjoe, Erasmus; Pawliszyn, Janusz

    2009-11-01

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

  18. 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.

  19. 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...

  20. 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...

  1. 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

  2. Dynamics of Microplates

    NASA Astrophysics Data System (ADS)

    Bodenschatz, E.; Katz, R.

    2005-12-01

    Rotating, growing microplates are observed in a wax analog model of sea-floor spreading. We show that wax microplates are kinematically similar to sea-floor tectonic microplates in terms of spreading rate, growth rate and their characteristic spiral pseudofault geometry. We propose a theory for the formation of microplates.

  3. 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

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

    PubMed

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

    2012-07-01

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

  5. 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

  6. Tectonic microplates: laying it down on wax

    NASA Astrophysics Data System (ADS)

    Katz, R. R.; Bodenschatz, E.

    2008-12-01

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

  7. 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.

  8. Automation of in-tip solid-phase microextraction in 96-well format for the determination of a model drug compound in human plasma by liquid chromatography with tandem mass spectrometric detection.

    PubMed

    Xie, W; Mullett, W M; Miller-Stein, C M; Pawliszyn, J

    2009-02-01

    Studies using in-tip solid phase microextraction (in-tip SPME) in a 96-well plate format are conducted to investigate the feasibility of SPME automation. The sample preparation process, including extraction and desorption, was fully automated and coupled with currently commercially available automated liquid handling systems. Several process parameters including extraction time and speed, and desorption time were investigated. An LC-MS/MS method has been developed and validated to determine the levels of a drug compound (MK-0533) in human plasma that demonstrates the suitability of this new approach. The developed method has a lower limit of quantitation (LLOQ) of 5 ng/mL when 0.25 mL of human plasma is processed and is validated in the concentration range of 5-2, 000 ng/mL. The successful application of the assay in clinical sample analysis indicates that in-tip SPME can be easily automated and has great potential to be used for high throughput quantitative determination of drugs in pharmaceutical industry.

  9. 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

  10. 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.

  11. 96-well microtiter plates for biofouling simulation in biomedical settings.

    PubMed

    Gomes, L C; Moreira, J M R; Teodósio, J S; Araújo, J D P; Miranda, J M; Simões, M; Melo, L F; Mergulhão, F J

    2014-01-01

    Microtiter plates with 96 wells are routinely used in biofilm research mainly because they enable high-throughput assays. These platforms are used in a variety of conditions ranging from static to dynamic operation using different shaking frequencies and orbital diameters. The main goals of this work were to assess the influence of nutrient concentration and flow conditions on biofilm formation by Escherichia coli in microtiter plates and to define the operational conditions to be used in order to simulate relevant biomedical scenarios. Assays were performed in static mode and in incubators with distinct orbital diameters using different concentrations of glucose, peptone and yeast extract. Computational fluid dynamics (CFD) was used to simulate the flow inside the wells for shaking frequencies ranging from 50 to 200 rpm and orbital diameters from 25 to 100 mm. Higher glucose concentrations enhanced adhesion of E. coli in the first 24 h, but variation in peptone and yeast extract concentration had no significant impact on biofilm formation. Numerical simulations indicate that 96-well microtiter plates can be used to simulate a variety of biomedical scenarios if the operating conditions are carefully set.

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

    PubMed

    Bharate, Sonali S; Vishwakarma, Ram A

    2015-04-01

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

  13. Microplate well coverage mixing using superhydrophobic contact.

    PubMed

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

    2012-11-01

    Two important challenges in microplate instrumentation are to achieve full well sample coverage and complete mixing. An effective approach of using superhydrophobic rods to accomplish these challenges is reported here. Experiments conducted showed that analytes above 50μl could be made to completely cover the bottom of 96-well standard and transparency microplates. Complete mixing was accomplished by moving the rod parallel to the well bottom while contacting the liquid. The approach is simple and controlled, and it minimizes the problems of spillage and cross-contamination. It works with analytes with varied volumes and of different viscosities present in each well of the microplate. PMID:22864233

  14. 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

  15. 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

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

    PubMed

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

    2016-01-01

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

  17. Quantitation of serrapeptase in formulations by UV method in the microplate format.

    PubMed

    Sandhya, K V; Devi, S Gayathri; Mathew, Sam T

    2008-10-01

    Serrapeptase is an anti-inflammatory, proteolytic enzyme isolated from the microorganism, Serratia sp. HY-6. Very few methods are available for the quantification of serrapeptase. The activity of the enzyme is determined by an ELISA assay, colorimetric method using casein as substrate or by HPLC method. These methods are lengthy, time consuming and require a number of reagents and solvents. Therefore an attempt was made to develop a simple alternative method for regular estimation of drug in formulations. Serrapeptase enzyme was estimated in formulations by using microplate readers which uses the principle of vertical photometry. Further this method was validated and the robustness of this method was checked by estimating the drug in various formulations including liposomes and marketed tablet formulations. A linear relationship between drug concentration and absorbance was observed between 1-4 microg/ml at 230 nm (R(2)=0.9911). The percentage recovery values of the drug in serrapeptase liposomes were found to lie within the standard limit (97-98%) which confirms the method is accurate and free from any positive or negative interference of the excipient. The low value of standard deviation obtained confirms the precision of the method. (+/-0.020 - +/-0.044). The drug content values in marketed tablets values obtained matched the label claim. The proposed microplate UV-method for determination of serrapeptase in formulations is novel, simple, inexpensive, fast, specific and robust. Thus this method could be a better alternative for regular estimation of drug in the various marketed formulations of serrapeptase.

  18. Parabolic growth patterns in 96-well plate cell growth experiments.

    PubMed

    Faessel, H M; Levasseur, L M; Slocum, H K; Greco, W R

    1999-05-01

    In preparing for the routine use of the ubiquitous in vitro cell growth inhibition assay for the study of anticancer agents, we characterized the statistical properties of the assay and found some surprising results. Parabolic well-to-well cell growth patterns were discovered, which could profoundly affect the results of routine growth inhibition studies of anticancer and other agents. Four human ovarian cell lines, A2780/WT, A2780/DX5, A2780/DX5B, and A121, and one human ileocecal adenocarcinoma cell line, HCT-8, were seeded into plastic 96-well plates with a 12-channel pipette, without drugs, and grown from 1-5 d. The wells were washed with a plate washer, cells stained with sulforhodamine B (SRB), and dye absorbance measured with a plate reader. Variance models were fit to the data from replicates to determine the nature of the heteroscedastic error structure. Exponential growth models were fit to data to estimate doubling times for each cell line. Polynomial models were fit to data from 10-plate stacks of 96-well plates to explore nonuniformity of cell growth in wells in different regions of the stacks. Each separate step in the assay was examined for precision, patterns, and underlying causes of variation. Differential evaporation of water from wells is likely a major, but not exclusive, contributor to the systematic well-to-well cell growth patterns. Because the fundamental underlying causes of the parabolic growth patterns were not conclusively found, a randomization step for the growth assay was developed.

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

    PubMed

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

    2015-01-01

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

  20. Fluorinert, an oxygen carrier, improves cell culture performance in deep square 96-well plates by facilitating oxygen transfer.

    PubMed

    Meyer, Aaron; Condon, Russell G G; Keil, Gregory; Jhaveri, Nikita; Liu, Zhong; Tsao, Yung-Shyeng

    2012-01-01

    In bioprocess development, the 96-well plate format has been widely used for high-throughput screening of production cell line or culture conditions. However, suspension cell cultures in conventional 96-well plates often fail to reach high cell density under normal agitation presumably due to constraints in oxygen transfer. Although more vigorous agitation can improve gas transfer in 96-well plate format, it often requires specialized instruments. In this report, we employed Fluorinert, a biologically inert perfluorocarbon, to improve oxygen transfer in 96-well plate and to enable the growth of a Chinese Hamster Ovary cell line expressing a recombinant monoclonal antibody. When different amounts of Fluorinert were added to the cell culture medium, a dose-dependent improvement in cell growth was observed in both conventional and deep square 96-well plates. When sufficient Fluorinert was present in the culture, the cell growth rate, the peak cell density, and recombinant protein production levels achieved in deep square 96-wells were comparable to cultures in ventilated shake flasks. Although Fluorinert is known to dissolve gases such as oxygen and CO(2), it does not dissolve nor extract medium components, such as glucose, lactate, or amino acids. We conclude that mixing Fluorinert with culture media is a suitable model for miniaturization of cell line development and process optimization. Proper cell growth and cellular productivity can be obtained with a standard shaker without the need for any additional aeration or vigorous agitation. PMID:21954223

  1. 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…

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

    NASA Astrophysics Data System (ADS)

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

    2016-03-01

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

  3. 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…

  4. A high-throughput, homogeneous microplate assay for agents that kill mammalian tissue culture cells.

    PubMed

    Pierce, Michael; Wang, Chunwei; Rebentisch, Matt; Endo, Mark; Stump, Mark; Kamb, Alexander

    2003-06-01

    Screens for cytostasis/cytoxicity have considerable value for the discovery of therapeutic agents and the investigation of the biology of apoptosis. For instance, genetic screens for proteins, protein fragments, peptides, RNAs, or chemicals that kill tissue culture cells may aid in identifying new cancer therapeutic targets. A microplate assay for cell death is needed to achieve throughputs sufficient to sift through thousands of agents from expression or chemical libraries. The authors describe a homogeneous assay for cell death in tissue culture cells compatible with 96- or 384-well plates. In combination with a previously described system for retroviral packaging and transduction, nearly 6000 expression library clones could be screened per week in a 96-well plate format. The screening system may also prove useful for chemical screens.

  5. Actively evolving microplate formation by oblique collision and sideways motion along strike-slip faults: An example from the northeastern Caribbean plate margin

    NASA Astrophysics Data System (ADS)

    Mann, Paul; Taylor, F. W.; Edwards, R. Lawrence; Ku, Teh-Lung

    1995-06-01

    The pattern of folding, faulting, and late Quaternary coral-reef uplift rates in western and central Hispaniola (Haiti and Dominican Republic) suggest that the elongate Gonave microplate, a 190,000-km 2 area of the northeastern Caribbean plate, is in the process of shearing off the Caribbean plate and accreting to the North American plate. Late Cenozoic transpression between the southeastern Bahama Platform and the Caribbean plate in Hispaniola has inhibited the eastward motion of the northeastern corner of the plate. Transpression is manifested in western and central Hispaniola by the formation of regional scale folds that correspond to present-day, anticlinal topographic mountain chains continuous with offshore anticlinal ridges. Areas of most rapid Quaternary uplift determined from onland coral reefs 125 ka and younger, coincide with the axial traces of these folds. Offshore data suggest recent folding and faulting of the seafloor. Onshore reef data do not conclusively require late Quaternary folding, but demonstrate that tectonic uplift rates of the axial areas of the anticlines decrease from the Northwest Peninsula of Haiti (0.37 mm/yr) to to the central part of the coast of western Haiti (0.19 mm/yr) to the south-central part of western Haiti (0 mm/yr). Formation of the 1200-km-long Enriquillo-Plantain Garden-Walton fault zone as a 'bypass' strike-slip fault has isolated the southern edge of the Gonave microplate and is allowing continued, unimpeded eastward motion of a smaller Caribbean plate past the zone of late Neogene convergence and Quaternary uplift of coral reefs in Hispaniola. Offshore seismic reflection data from the Jamaica Passage, the marine strait separating Jamaica and Haiti, show that the Enriquillo-Plantain Garden fault zone forms a narrow but deep, active fault-bounded trough beneath the passage. The active fault is continuous with active faults mapped onshore in western Haiti and eastern Jamaica; the bathymetric deep is present because the

  6. 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

  7. A suite of microplate reader-based colorimetric methods to quantify ammonium, nitrate, orthophosphate and silicate concentrations for aquatic nutrient monitoring.

    PubMed

    Ringuet, Stephanie; Sassano, Lara; Johnson, Zackary I

    2011-02-01

    A sensitive, accurate and rapid analysis of major nutrients in aquatic systems is essential for monitoring and maintaining healthy aquatic environments. In particular, monitoring ammonium (NH(4)(+)) concentrations is necessary for maintenance of many fish stocks, while accurate monitoring and regulation of ammonium, orthophosphate (PO(4)(3-)), silicate (Si(OH)(4)) and nitrate (NO(3)(-)) concentrations are required for regulating algae production. Monitoring of wastewater streams is also required for many aquaculture, municipal and industrial wastewater facilities to comply with local, state or federal water quality effluent regulations. Traditional methods for quantifying these nutrient concentrations often require laborious techniques or expensive specialized equipment making these analyses difficult. Here we present four alternative microcolorimetric assays that are based on a standard 96-well microplate format and microplate reader that simplify the quantification of each of these nutrients. Each method uses small sample volumes (200 µL), has a detection limit ≤ 1 µM in freshwater and ≤ 2 µM in saltwater, precision of at least 8% and compares favorably with standard analytical procedures. Routine use of these techniques in the laboratory and at an aquaculture facility to monitor nutrient concentrations associated with microalgae growth demonstrates that they are rapid, accurate and highly reproducible among different users. These techniques offer an alternative to standard nutrient analyses and because they are based on the standard 96-well format, they significantly decrease the cost and time of processing while maintaining high precision and sensitivity.

  8. Transparency-based microplates for fluorescence quantification.

    PubMed

    Cheong, Brandon Huey-Ping; Diep, Vu; Ng, Tuck Wah; Liew, Oi Wah

    2012-03-01

    Microplates for use in resource-limited laboratories should ideally not require processes that involve substantial large-scale production in order to be viable. We describe and demonstrate here an approach of using a silicone sheet with holes, conveniently cut out precisely using an inexpensive cutting plotter to correspond with regions where liquid is to be dispensed, and attaching it to a transparency to create very thin well arrays. With this, the contact angle hysteresis behavior of liquid could be harnessed to produce taller drop shapes so that the fiber probe used could read in the emitted light more effectively. Experimentation conducted revealed fluorescence measurements that were significantly more sensitive than standard microplates, notwithstanding that smaller volumes of liquid were needed. This was achieved using both the fiber optic and imaging evaluation modes. The two methods investigated, one with a lid placed and one without, showed the latter to produce marginally more sensitive readings as opposed to improved immunity from the environment with the former. These favorable measurement characteristics were found to be achievable with an estimated production cost of AU $0.40 and fabrication times of 3.5 min (96 wells) and 6.5 min (384 wells) per plate. PMID:22266206

  9. Macroscale versus microscale methods for physiological analysis of biofilms formed in 96-well microtiter plates.

    PubMed

    Gomes, L C; Moreira, J M R; Miranda, J M; Simões, M; Melo, L F; Mergulhão, F J

    2013-12-01

    Microtiter plates with 96 wells have become one of the preferred platforms for biofilm studies mainly because they enable high-throughput assays. In this work, macroscale and microscale methods were used to study the impact of hydrodynamic conditions on the physiology and location of Escherichia coli JM109(DE3) biofilms formed in microtiter plates. Biofilms were formed in shaking and static conditions, and two macroscale parameters were assayed: the total amount of biofilm was measured by the crystal violet assay and the metabolic activity was determined by the resazurin assay. From the macroscale point of view, there were no statistically significant differences between the biofilms formed in static and shaking conditions. However, at a microscale level, the differences between both conditions were revealed using scanning electron microscopy (SEM). It was observed that biofilm morphology and spatial distribution along the wall were different in these conditions. Simulation of the hydrodynamic conditions inside the wells at a microscale was performed by computational fluid dynamics (CFD). These simulations showed that the shear strain rate was unevenly distributed on the walls during shaking conditions and that regions of higher shear strain rate were obtained closer to the air/liquid interface. Additionally, it was shown that wall regions subjected to higher shear strain rates were associated with the formation of biofilms containing cells of smaller size. Conversely, regions with lower shear strain rate were prone to have a more uniform spatial distribution of adhered cells of larger size. The results presented on this work highlight the wealth of information that may be gathered by complementing macroscale approaches with a microscale analysis of the experiments. PMID:24140575

  10. 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...

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

    PubMed Central

    Quah, Joanne; Tong, Louis; Kim, Namyong

    2015-01-01

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

  12. 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.

  13. 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.

  14. 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

  15. Quantification of lymphocytic choriomeningitis virus with an immunological focus assay in 24- or 96-well plates.

    PubMed

    Battegay, M; Cooper, S; Althage, A; Bänziger, J; Hengartner, H; Zinkernagel, R M

    1991-06-01

    Titers of lymphocytic choriomeningitis virus (LCMV) were determined on adherent fibroblast cell lines in 24- or 96-well plates. After absorption of virus by cells and 48 h incubation under a methylcellulose overlay, cell monolayers were fixed with 4% formaldehyde in phosphate-buffered saline, permeabilized by incubation in 0.5% Triton X-100 in balanced salt solution and then stained with a monoclonal rat anti-LCMV and a peroxidase-labeled second stage antibody. The sensitivity of the assay is within a factor of 2-4 of conventional plaquing methods. The method also detects poorly or non-plaquing LCMV isolates, and therefore drastically reduces the need for titration of LCMV in mice. The method is quicker (2-3 days), as compared to conventional methods (4-6 days) and less expensive in terms of work and materials.

  16. 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.

  17. A solid-phase Bcr-Abl kinase assay in 96-well hydrogel plates.

    PubMed

    Wu, Ding; Mand, Michael R; Veach, Darren R; Parker, Laurie L; Clarkson, Bayard; Kron, Stephen J

    2008-04-01

    Regulated phosphorylation by protein tyrosine kinases (PTKs), such as c-Abl, is critical to cellular homeostasis. In turn, once deregulated as in the chronic myeloid leukemia (CML) fusion protein Bcr-Abl, PTKs can promote cancer onset and progression. The dramatic success of the Bcr-Abl inhibitor imatinib as therapy for CML has inspired interest in other PTKs as targets for cancer drug discovery. Here we report a novel PTK activity and inhibition screening method using hydrogel-immobilized peptide substrates. Using acrylate crosslinkers, we tether peptides via terminal cysteines to thiol-presenting hydrogels in 96-well plates. These surfaces display low background and high reproducibility, allowing semiquantitative detection of peptide phosphorylation by recombinant c-Abl or by Bcr-Abl activity in cell extracts using traditional anti-phosphotyrosine immunodetection and chemifluorescence. The capabilities of this assay are demonstrated by performing model screens for inhibition with several commercially available PTK inhibitors and a collection of pyridopyrimidine Src/Abl dual inhibitors. This assay provides a practical method to measure the activity of a single kinase present in a whole cell lysate with high sensitivity and specificity as a valuable means for efficient small molecule screening.

  18. Development of a quantitative 96-well method to image glycogen storage in primary rat hepatocytes.

    PubMed

    Pilling, James; Garside, Helen; Ainscow, Edward

    2010-08-01

    Within the liver, hormonal control of glycogen metabolism allows for rapid release and uptake of glucose from the circulation, providing a reserve of glucose that can be utilised by other organs. Traditionally, cellular glycogen storage has been detected using Periodic acid Schiff (PAS) staining of histopathology samples or a biochemical assay. Colorimetric measurement of glycogen content using PAS staining is hard to quantify whilst biochemical techniques give limited information about events such as cytotoxicity or allow analysis of hepatic heterogeneity. Here, we describe the development of an imaging based method to quantify glycogen storage in 96-well cultures of primary rat hepatocytes using the inherent fluorescence properties of the Schiff reagent. PAS-stained hepatocytes were imaged using an automated fluorescent microscope, with the amount of glycogen present in each cell being quantified. Using this technique, we found an increase in glycogen storage in response to insulin (EC50 = 0.31 nM) that was in agreement with that determined using biochemical quantification (EC50 = 0.32 nM). Furthermore, a dose dependent increase in glycogen storage was also seen in response to glycogen synthase kinase inhibitors and glycogen phosphorylase inhibitors. This technique allows rapid assessment of cellular glycogen storage in response to hormones and small molecule inhibitors.

  19. CometChip: a high-throughput 96-well platform for measuring DNA damage in microarrayed human cells.

    PubMed

    Ge, Jing; Prasongtanakij, Somsak; Wood, David K; Weingeist, David M; Fessler, Jessica; Navasummrit, Panida; Ruchirawat, Mathuros; Engelward, Bevin P

    2014-10-18

    DNA damaging agents can promote aging, disease and cancer and they are ubiquitous in the environment and produced within human cells as normal cellular metabolites. Ironically, at high doses DNA damaging agents are also used to treat cancer. The ability to quantify DNA damage responses is thus critical in the public health, pharmaceutical and clinical domains. Here, we describe a novel platform that exploits microfabrication techniques to pattern cells in a fixed microarray. The 'CometChip' is based upon the well-established single cell gel electrophoresis assay (a.k.a. the comet assay), which estimates the level of DNA damage by evaluating the extent of DNA migration through a matrix in an electrical field. The type of damage measured by this assay includes abasic sites, crosslinks, and strand breaks. Instead of being randomly dispersed in agarose in the traditional assay, cells are captured into an agarose microwell array by gravity. The platform also expands from the size of a standard microscope slide to a 96-well format, enabling parallel processing. Here we describe the protocols of using the chip to evaluate DNA damage caused by known genotoxic agents and the cellular repair response followed after exposure. Through the integration of biological and engineering principles, this method potentiates robust and sensitive measurements of DNA damage in human cells and provides the necessary throughput for genotoxicity testing, drug development, epidemiological studies and clinical assays.

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

    NASA Technical Reports Server (NTRS)

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

    1986-01-01

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

  1. 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. 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

  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%.

  6. 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

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

    PubMed

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

    2009-03-01

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

  8. Seismicity of the Adriatic microplate

    USGS Publications Warehouse

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

    1993-01-01

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

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

    PubMed

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

    2006-01-01

    A novel 96-well screen filter plate (patent pending) has been invented to eliminate a time-consuming and labor-intensive step in preparation of in vivo study samples--to remove blood or plasma clots. These clots plug the pipet tips during a manual or automated sample-transfer step causing inaccurate pipetting or total pipetting failure. Traditionally, these blood and plasma clots are removed by picking them out manually one by one from each sample tube before any sample transfer can be made. This has significantly slowed the sample preparation process and has become a bottleneck for automated high-throughput sample preparation using robotic liquid handlers. Our novel screen filter plate was developed to solve this problem. The 96-well screen filter plate consists of 96 stainless steel wire-mesh screen tubes connected to the 96 openings of a top plate so that the screen filter plate can be readily inserted into a 96-well sample storage plate. Upon insertion, the blood and plasma clots are excluded from entering the screen tube while clear sample solutions flow freely into it. In this way, sample transfer can be easily completed by either manual or automated pipetting methods. In this report, three structurally diverse compounds were selected to evaluate and validate the use of the screen filter plate. The plasma samples of these compounds were transferred and processed in the presence and absence of the screen filter plate and then analyzed by LC-MS/MS methods. Our results showed a good agreement between the samples prepared with and without the screen filter plate, demonstrating the utility and efficiency of this novel device for preparation of blood and plasma samples. The device is simple, easy to use, and reusable. It can be employed for sample preparation of other biological fluids that contain floating particulates or aggregates. PMID:16383347

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

    PubMed

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

    2011-11-09

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

  11. 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

  12. Optical microplates for high-throughput screening of photosynthesis in lipid-producing algae.

    PubMed

    Chen, Meng; Mertiri, Taulant; Holland, Thomas; Basu, Amar S

    2012-10-21

    It is well known that biological systems respond to chemical signals as well as physical stimuli. The workhorses of high throughput screening, microplates and pipetting robots, are well suited for screening chemical stimuli; however, there are fewer options for screening physical stimuli, particularly those which involve temporal patterns. This paper presents an optical microplate for photonic high-throughput screening. The system provides addressable intensity and temporal control of LED light emission in each well, and operates on standard black-wall clear-bottom 96-well microplates, which prevent light spillover. Light intensity can be controlled to 7-bit resolution (128 levels), with a maximum intensity of 120 mE cm(-2). The temporal resolution, useful for studying dynamics of light-driven bioprocesses, can be as low as 10 μs. The microplate is used for high-throughput studies of light-dependent growth rates and photosynthetic efficiency in the model organism Dunaliella tertiolecta, a lipid-producing algae of interest in 2(nd) generation biofuels. By conducting 96 experiments in parallel, photoirradiance studies, which would require 2 years using conventional tools, can be completed in <2 weeks. In a 12 day culture, algal growth rates increase with total photon flux, as expected. Interestingly, the lipid production efficiency, defined as lipid production per unit photon flux per capita, increases nearly 5 fold at low light intensity (constant light) and at low duty cycle (pulsed light). High throughput protocols enabled by this system are conducive to systematic studies and discovery in the fields of photobiology and photochemistry.

  13. 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

  14. CYP isoform induction screening in 96-well plates: use of 7-benzyloxy-4-trifluoromethylcoumarin as a substrate for studies with rat hepatocytes.

    PubMed

    Price, R J; Surry, D; Renwick, A B; Meneses-Lorente, G; Lake, B G; Evans, D C

    2000-08-01

    1. In this study, 7-benzyloxy-4-trifluoromethylcoumarin (BFC) was evaluated as a substrate to assess the induction of cytochrome P450 (CYP) isoform enzyme activities in rat hepatocytes using a 96-well plate format. 2. BFC was metabolized by both untreated and sodium phenobarbitone (NaPB)-treated rat hepatocytes in a time- and concentration-dependent manner to the highly fluorescent product 7-hydroxy-4-trifluoromethylcoumarin (HFC). 3. HFC was extensively conjugated with D-glucuronic acid and/or sulphate in both untreated and NaPB-treated rat hepatocytes, thus necessitating the inclusion of an enzymatic deconjugation step in the assay procedure. 4. The time-course of induction of 7-ethoxyresorufin metabolism by the CYP1A inducer beta-naphthoflavone (BNF), 7-benzyloxyresorufin metabolism by the CYP2B inducer NaPB and BFC metabolism b both BNF and NaPB was studied in rat hepatocytes treated for 24-96 h. The optimal time for induction of metabolism of all three substrates was 72 h, with no medium changes being necessary during this period. 5. The effect of treatment with 0.5-20 microM BNF, 50-2000 microM NaPB, 2-20 microM dexamethasone (DEX), 20-100 microM methylclofenapate (MCP), and 50 and 200 microM isoniazid (ISN) for 72 h on BFC metabolism in cultured rat hepatocytes was studied. BFC metabolism was induced by treatment with BNF, NaPB and MCP, but not with either DEX or ISN. 6. The metabolism of BFC in liver microsomes from the control rat and rat treated with CYP isoform inducers was also studied. BFC metabolism was induced by treatment with NaPB, BNF and DEX. 7. The metabolism of BFC was also studied using microsomes from baculovirus-infected insect cells containing rat cDNA-expressed CYP1A, CYP2B, CYP2C and CYP3A isoforms. Whereas BFC was metabolized to some extent by all the rat cDNA-expressed CYP isoforms examined, at a substrate concentration of 2.5 microM the greatest rates of BFC metabolism were observed with the CYP1A1, CYP1A2 and CYP2B1 preparations. 8

  15. Philippine microplate tectonics and hydrocarbon exploration

    SciTech Connect

    Gallagher, J.J. Jr.

    1986-07-01

    Hydrocarbon traps in the Philippine Islands developed during a long, complex history of microplate tectonics. Carbonate and clastic stratigraphic traps formed during Mesozoic and early Cenozoic rifting and drifting. Hydrocarbons, generated in deep rift basins, migrated to the traps during drifting. Later Cenozoic compressional tectonic activity and concomitant faulting enhanced some traps and destroyed others. Seismic data offshore from Palawan Island reveal the early trap histories. Later trap histories can be interpreted from seismic, outcrop, and remote-sensing data. Understanding the microplate tectonic history of the Philippines is the key to interpreting trap histories.

  16. A simple 96 well microfluidic chip combined with visual and densitometry detection for resource-poor point of care testing

    PubMed Central

    Yang, Minghui; Sun, Steven; Kostov, Yordan

    2010-01-01

    There is a well-recognized need for low cost biodetection technologies for resource-poor settings with minimal medical infrastructure. Lab-on-a-chip (LOC) technology has the ability to perform biological assays in such settings. The aim of this work is to develop a low cost, high-throughput detection system for the analysis of 96 samples simultaneously outside the laboratory setting. To achieve this aim, several biosensing elements were combined: a syringe operated ELISA lab-on-a-chip (ELISA-LOC) which integrates fluid delivery system into a miniature 96-well plate; a simplified non-enzymatic reporter and detection approach using a gold nanoparticle-antibody conjugate as a secondary antibody and silver enhancement of the visual signal; and Carbon nanotubes (CNT) to increase primary antibody immobilization and improve assay sensitivity. Combined, these elements obviate the need for an ELISA washer, electrical power for operation and a sophisticated detector. We demonstrate the use of the device for detection of Staphylococcal enterotoxin B, a major foodborne toxin using three modes of detection, visual detection, CCD camera and document scanner. With visual detection or using a document scanner to measure the signal, the limit of detection (LOD) was 0.5ng/ml. In addition to visual detection, for precise quantitation of signal using densitometry and a CCD camera, the LOD was 0.1ng/ml for the CCD analysis and 0.5 ng/ml for the document scanner. The observed sensitivity is in the same range as laboratory-based ELISA testing. The point of care device can analyze 96 samples simultaneously, permitting high throughput diagnostics in the field and in resource poor areas without ready access to laboratory facilities or electricity. PMID:21503269

  17. 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

  18. 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

  19. Counter-rotating microplates at the Galapagos triple junction.

    PubMed

    Klein, Emily M; Smith, Deborah K; Williams, Clare M; Schouten, Hans

    2005-02-24

    An 'incipient' spreading centre east of (and orthogonal to) the East Pacific Rise at 2 degrees 40' N has been identified as forming a portion of the northern boundary of the Galapagos microplate. This spreading centre was described as a slowly diverging, westward propagating rift, tapering towards the East Pacific Rise. Here we present evidence that the 'incipient rift' has also rifted towards the east and opens anticlockwise about a pivot at its eastern end. The 'incipient rift' then bounds a second microplate, north of the clockwise-rotating Galapagos microplate. The Galapagos triple junction region, in the eastern equatorial Pacific Ocean, thus consists of two counter-rotating microplates partly separated by the Hess Deep rift. Our kinematic solution for microplate motion relative to the major plates indicates that the two counter-rotating microplates may be treated as rigid blocks driven by drag on the microplates' edges3.

  20. 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

  1. Easter microplate evolution: Pb isotope evidence

    NASA Astrophysics Data System (ADS)

    Hanan, Barry B.; Schilling, Jean-Guy

    1989-06-01

    We report on 53 Pb isotope analyses of basalts from 48 dredge stations occupied along the spreading boundaries of the Easter microplate and adjacent East Pacific Rise (EPR). Also included in the study are seven analyses of basalts from Easter and Sala y Gomez islands. A major anomaly is observed on the East Rift, around 27°S, where this ridge is shallowest and closest to Easter and Sala y Gomez islands. Basalts from the West Rift are less radiogenic. The means for the two rift populations are distinct, but their ranges overlap significantly. On the average, there is a systematic westward decrease in radiogenic Pb content with distance from Sala y Gomez. The Pb isotope anomaly is confined to the boundaries of the Microplate and the total range exceeds that of the entire EPR, both in the most and the least end of radiogenic Pb content. Radiogenic Pb content covaries with (La/Sm)N ratios with the exception of a nepheline-normative picritic basalt glass outlier. The trends are curvilinear. There is no correlation between the Pb isotope ratios and the bulk composition of the lavas. In Pb versus Pb isotope diagrams, basalts from the East and West rifts form two tight linear trends of statistically indistinguishable slope. Basalts from Easter and Sala y Gomez islands lie on the upper end of these trends. A binary mixing process between a radiogenic source similar to that present beneath Sala y Gomez and the large ion lithophile element (LILE)-depleted mid-ocean ridge basalt (MORB) source is strongly suggested. There is no trace of the Dupal anomaly beneath the microplate nor beneath Easter or Sala y Gomez Islands. If the Dupal anomaly is indeed continuous and of semi global extent, it must lie deeper in the mantle than the depths at which basaltic melts take source beneath the microplate and these two islands. There is also no correlation between the apparent dispersion of Pb isotope ratios and the rate at which the various ridge segments of the microplate spread. Tests

  2. Surface-scribed transparency-based microplates.

    PubMed

    Li, Xin Ye; Cheong, Brandon Huey-Ping; Somers, Anthony; Liew, Oi Wah; Ng, Tuck Wah

    2013-01-15

    Transparency sheets, which are normally associated with use on overhead projectors, offer lowered costs and high amenability for optical diagnostics in microplate instrumentation. An alternative microplate design in which circles are scribed on the surface of the transparency to create the boundaries to hold the drop in place is investigated here. The 3D profile of the scribed regions obtained optically showed strong likelihood of affecting three-phase contact line interactions. During dispensation, the contact angle (≈95°) was larger than the drop advancing state (≈80°) due to a period of nonadhesion, where the contact angle later reduced to the drop advancing state followed by increase in the liquid area coverage on the substrate. It was established that 50 μL was needed to fill the well fully, and the maximum volume retainable before breaching was 190 μL. While the tilt angle needed for displacement reduced significantly from 50 to 95 μL, this was markedly better than nonscribed surfaces, where tilt angles always had to be kept to within 30°. It was found that there was greater ability to fill the well with smaller volumes with dispensation at the center. This was attributed to the growing contact line not meeting the scribed edge in parallel if liquid was dispensed closer to it, wherein pinning reduction in some directions permitted liquid travel along the scribed edge to undergo contact angle hysteresis. Fluorescence measurements conducted showed no performance compromise when using scribed transparency microplates over standard microplates. PMID:23215012

  3. 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

  4. 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

  5. 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.

  6. Measurement of factor v activity in human plasma using a microplate coagulation assay.

    PubMed

    Tilley, Derek; Levit, Irina; Samis, John A

    2012-09-09

    In response to injury, blood coagulation is activated and results in generation of the clotting protease, thrombin. Thrombin cleaves fibrinogen to fibrin which forms an insoluble clot that stops hemorrhage. Factor V (FV) in its activated form, FVa, is a critical cofactor for the protease FXa and accelerator of thrombin generation during fibrin clot formation as part of prothrombinase (1, 2). Manual FV assays have been described (3, 4), but they are time consuming and subjective. Automated FV assays have been reported (5-7), but the analyzer and reagents are expensive and generally provide only the clot time, not the rate and extent of fibrin formation. The microplate platform is preferred for measuring enzyme-catalyzed events because of convenience, time, cost, small volume, continuous monitoring, and high-throughput (8, 9). Microplate assays have been reported for clot lysis (10), platelet aggregation (11), and coagulation Factors (12), but not for FV activity in human plasma. The goal of the method was to develop a microplate assay that measures FV activity during fibrin formation in human plasma. This novel microplate method outlines a simple, inexpensive, and rapid assay of FV activity in human plasma. The assay utilizes a kinetic microplate reader to monitor the absorbance change at 405 nm during fibrin formation in human plasma (Figure 1) (13). The assay accurately measures the time, initial rate, and extent of fibrin clot formation. It requires only μl quantities of plasma, is complete in 6 min, has high-throughput, is sensitive to 24-80 pM FV, and measures the amount of unintentionally activated (1-stage activity) and thrombin-activated FV (2-stage activity) to obtain a complete assessment of its total functional activity (2-stage activity - 1-stage activity). Disseminated intravascular coagulation (DIC) is an acquired coagulopathy that most often develops from pre-existing infections (14). DIC is associated with a poor prognosis and increases mortality

  7. 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.

  8. Present-day microplates motion in the central Mediterranean

    NASA Astrophysics Data System (ADS)

    D'Agostino, N.

    2007-12-01

    Motion of microplates in diffuse plate boundaries often accommodates convergence between major plates. In this work we present a kinematic model for the central mediterranean sector of the Eurasia-Nubia plate boundary, derived from GPS and earthquake slip vectors, which describe the crustal motion as resulting from the interactions of two rigid microplates: Adria and Apulia-Ionia. The styles of deformation predicted by the kinematic model along the boundaries of the proposed microplates are consistent with geological observations and present significant implications for the characterization of seismically active regions in the Eurasia-Nubia plate boundary. Microplates rotation rates are succesfully reproduced by a simple slat model in which the edges of the microplates are constrained to move with the adjacent plate or microplate, suggesting a significant coupling between crustal blocks and the primary role of edge-transmitted forces as driving factor for crustal motion. These results are compared with surface wave tomographic models and other seismological observables which suggest that the rigid microplates correspond to regions of high shear wave velocities in the upper mantle and high integrated lithospheric strength. The present-day plate boundary configuration derives from the inherited Mesozoic African margin, from the rapid Neogene evolution of the Tyrrhenian subduction zone and the final fragmentation of the Adriatic promontory.

  9. Tectonic microplates in a wax model of sea-floor spreading

    NASA Astrophysics Data System (ADS)

    Katz, Richard F.; Ragnarsson, Rolf; Bodenschatz, Eberhard

    2005-01-01

    Rotating, growing microplates are observed in a wax analogue model of sea-floor spreading. Wax microplates are kinematically similar to sea-floor tectonic microplates in terms of spreading rate and growth rate. Furthermore, their spiral pseudofault geometry is quantitatively consistent with Schouten's oceanic microplate model. These results suggest that Schouten's edge-driven microplate model captures the kinematics of tectonic microplate evolution on Earth. Based on the wax observations, a theory for the nucleation of overlapping spreading centres, the precursors of tectonic microplates, is developed.

  10. 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

  11. Single laboratory method performance evaluation for the analysis of total food folate by trienzyme extraction and microplate assay.

    PubMed

    Chen, L; Eitenmiller, R R

    2007-06-01

    Single laboratory method performance parameters, including the calibration curve, accuracy, recovery, precision, limit of detection (LOD), and limit of quantification (LOQ), were evaluated for the analysis of total food folate by the trienzyme extraction and microplate assay with Lactobacillus casei subsp. rhamnosus. Standard Reference Material (SRM) 1546 (meat homogenate), SRM 2383 (baby food composite), SRM 1846 (infant formula), Certified Reference Material (CRM) 121 (wholemeal flour), and CRM 485 (mixed vegetables), representing a broad selection of food matrices, were used to evaluate the performance of the method. A generated 4-parameter logistic equation of the calibration curve was y= (0.0705 - 1.0396)/(1 + (x/0.0165) (1.3072)) + 1.0396 (P < 0.0001). The test of parallelism demonstrated that matrix components in the food extracts did not affect the accuracy. Measured values of the SRMs and CRMs were within their certified or reference values. Recoveries for all reference materials met the requirements of the AOAC guidelines for single laboratory validation. Precision measured as repeatability, including simultaneous and consecutive replicates for each SRM and CRM, met the Horwitz criterion. LOD and LOQ values were 0.3 and 0.6 mug/100 g, respectively. The results showed that trienzyme digestion using alpha-amylase, Pronase(R), and conjugase from chicken pancreas coupled with a 96-well microplate assay provided a highly accurate, reproducible, and sensitive method for the determination of folate in a variety of foods.

  12. A microplate version of the SOS/umu-test for rapid detection of genotoxins and genotoxic potentials of environmental samples.

    PubMed

    Reifferscheid, G; Heil, J; Oda, Y; Zahn, R K

    1991-12-01

    The umu-microtest is a miniaturized automated short-term test version proposed for screening of umuC-dependent mutagenic potentials of chemicals relevant to environmental pollution, river water and industrial waste water. The test is based on the SOS/umu-test and has been modified in order to allow extensive testing of environmental samples. Genetically engineered Salmonella typhimurium (TA1535/pSK1002) are incubated on a microplate rotor in a sloping position for 2 h with the test samples, followed by addition of fresh culture medium to reach a 10-fold dilution of the incubation medium. 2 h later, the activity of the beta-galactosidase, which reflects umuC induction, is determined colorimetrically. The incubation of the bacteria in the presence of the test compounds as well as the assessment of beta-galactosidase activity takes place in 96-well microplates, thus enabling simultaneous screening of large numbers of samples. Data of the genotoxic potentials are available within 8 h. Computer-controlled automation is possible by using a laboratory workstation.

  13. Cell Treatment and Lysis in 96-Well Filter-Bottom Plates for Screening Bcr-Abl Activity and Inhibition in Whole-Cell Extracts

    PubMed Central

    MAND, MICHAEL R.; WU, DING; VEACH, DARREN R.; KRON, STEPHEN J.

    2015-01-01

    Although conventional high-throughput screens performed in vitro with purified protein kinases are powerful tools to discover new kinase inhibitors, they are far from ideal for determining efficacy in vivo. As a complementary approach, cell-based, target-driven secondary screens may help predict in vivo compound potency and specificity as well as evaluate bioavailability and toxicity. Here the authors report a simple protocol for treating K562 Bcr-Abl-expressing cells with small-molecule kinase inhibitors in 96-well filter-bottom plates followed by in-plate cell lysis. The lysates were assayed via a solid-phase kinase assay, allowing determination of apparent IC50 for known Bcr-Abl inhibitors as well as facilitating the screening of a small kinase inhibitor library. This approach may have further applications in generating lysates for analyzing kinase activity and inhibition in other nonadherent suspension cell lines. PMID:20237206

  14. Toward Biomarker Development in Large Clinical Cohorts: An Integrated High-Throughput 96-Well-Plate-Based Sample Preparation Workflow for Versatile Downstream Proteomic Analyses.

    PubMed

    Sun, Zeyu; Liu, Xiaoli; Jiang, Jing; Huang, Haijun; Wang, Jie; Wu, Daxian; Li, Lanjuan

    2016-09-01

    We describe a cheap, robust, fast, high-throughput, and flexible proteomic sample processing method based on a regular 96-well plate by acetone precipitation under low centrifuge speed (96PACS), which enables predigestion processing of 96 samples within 2 h. Tested on a complex Huh-7 total lysate, 96PACS produced comparable proteome coverage and even showed better reproducibility than FASP. Quantitative performance of 96PACS was further tested using data-independent acquisition and parallel reaction monitoring quantitation in a set of 6 benchmark samples consisting of 6 serial dilutions of BSA spiked in complex E. coli proteome background. The protocol was also successfully modified for automation and was validated in a comparative label-free proteomic study to develop serum markers for early detection of liver fibrosis and necroinflammation in patients chronically infected with hepatitis B virus. PMID:27471874

  15. 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

  16. Higher resolution microplate array diagonal gel electrophoresis: application to a multiallelic minisatellite.

    PubMed

    O'Dell, S D; Chen, X; Day, I N

    2000-01-01

    The 5' polymorphic region of the insulin (INS, MIM# 176730) gene contains a variable tandem repetition of 14-15 bp (a variable number of tandem repeats (VNTR) locus). After PCR amplification, we achieved precise sizing of class I alleles (range 641 to 843 bp) on 96-well open-face polyacrylamide microplate array diagonal gel electrophoresis (MADGE) gels, obtaining resolution of the 2% mobility difference which represents one tandem repeat. PCR products were run double-stranded, but no additional bands were generated except in the case of differences of three, two, and one repeat between alleles; none compromised allele identification, and in the latter case the heteroduplex was a useful confirmation signal. No end labelling of primers was required, as the sensitive Vistra Green intercalating dye for double strands was used for visualization of bands from diluted samples. Duracryl, a high mechanical-strength polyacrylamide derivative, proved to have good resolution properties for electrophoresis. A co-run ladder ensured precise binning without inter-lane variability. Simultaneous electrophoresis of gels in a thermostatically controlled tank allowed up to 1,000 samples to be run in 90 min. Gels were analyzed using a FluorImager 595 fluorescent scanning system, and alleles identified using a combination of Phoretix software for band migration measurement and Microsoft Excel to compute allele sizes. Unlike other systems for minisatellite allele sizing, throughput was not limited (in time or cost) by electrophoresis.

  17. Detection of sodium channel activators by a rapid fluorimetric microplate assay.

    PubMed

    Louzao, M C; Vieytes, M R; Yasumoto, T; Botana, L M

    2004-04-01

    Marine toxins such as brevetoxins and ciguatoxins are produced by dinoflagellates and can accumulate in seafood. These toxins affect humans through seafood consumption. Intoxication is mainly characterized by gastrointestinal and neurological disorders and, in most severe cases, by cardiovascular problems. To prevent the consumption of food contaminated with these toxins, shellfish have been tested by mouse bioassay. However, this method is expensive, time-consuming, and ethically questionable. The objective of this study was to use a recently developed fluorimetric microplate assay to rapidly detect brevetoxins and ciguatoxins. The method is based on the pharmacological effect of brevetoxins and ciguatoxins known to activate sodium channels and involves (i). the incubation of excitable cells in 96 well microtiter plates with the fluorescent dye bis-oxonol, whose distribution across the membrane is potential-dependent, and (ii). dose-dependent cell depolarization by the toxins. Our findings demonstrate that measuring changes in membrane potential induced by brevetoxins and ciguatoxins allowed their quantitation. Active toxins could be reliably detected at concentrations in the nanomolar range. The simplicity, sensitivity, and possibility of being automated provide the basis for development of a practical alternative to conventional testing for brevetoxins and ciguatoxins.

  18. Optical microplates for photonic high throughput screening of algal photosynthesis and biofuel production.

    PubMed

    Mertiri, Taulant; Chen, Meng; Holland, Thomas; Basu, Amar S

    2011-01-01

    Biological systems respond not only to chemical stimuli (drugs, proteins) but also to physical stimuli (light, heat, stress). Though there are many high throughput tools for screening chemical stimuli, no such tool exists for screening of physical stimuli. This paper presents a novel instrument for photonic high throughput screening of photosynthesis, a light-driven bioprocess. The optical microplate has a footprint identical to a standard 96 well plate, and it provides temporal and intensity control of light in each individual well. Intensity control provides 128 dimming levels (7-bit resolution), with maximum intensity 120 mE/cm(2). Temporal modulation, used for studying dynamics and regulation of photosynthesis, can be as low as 10 μs. We used photonic screening for high throughput studies of algal growth rates and photosynthetic efficiency, using the model organism Dunaliella tertiolecta, a lipid producing algae of interest in biofuel production. Due to the ability to conduct 96 studies in parallel, experiments that would require 2 years using conventional tools can be completed in 1 week. This instrument opens up novel high throughput protocols for photobiology and the growing field of phenomics.

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

    PubMed

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

    2002-05-01

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

  20. 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

  1. 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.

  2. 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

  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. 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

  5. Microwave-Accelerated Metal-Enhanced Fluorescence (MAMEF) with silver colloids in 96-well plates: Application to ultra fast and sensitive immunoassays, High Throughput Screening and drug discovery.

    PubMed

    Aslan, Kadir; Holley, Patrick; Geddes, Chris D

    2006-05-30

    Fluorescence detection is the basis of most assays used in drug discovery and High Throughput Screening (HTS) today. In all of these assays, assay rapidity and sensitivity is a primary concern, the sensitivity determined by both the quantum yield of the fluorophores and efficiency of the detection system, while rapidity is determined by the physical and biophysical parameters of temperature, concentration, assay bioaffinity, etc. In this paper we describe a platform technology that promises to fundamentally address these two physical constraints of sensitivity and rapidity. By combining the use of Metal-Enhanced Fluorescence (MEF), a near-field effect that can significantly enhance fluorescence signatures, with low power microwave heating, we can significantly increase the sensitivity of surface assays as well as >95% kinetically complete the assay within a few seconds. In addition, the metallic nanostructures used to facilitate MEF appear to be preferentially heated as compared to the surface assay fluid, advantageously localizing the MEF and heating around the nanostructures. To demonstrate proof of principle, a 96-well plate has been functionalized with silver nanostructures, and a model protein avidin-biotin assay studied. In our findings, a greater than 5-fold fluorescence enhancement coupled with a approximately 90-fold increase in assay kinetics was observed, but with no assay washing steps needed due to the silver-enhanced evanescent field mode of excitation. These findings promise to strongly facilitate high throughput fluorescence-based processes, such as in biology, drug discovery and general compound screening.

  6. 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.

  7. Absorbance and fluorometric sensing with capillary wells microplates.

    PubMed

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

    2010-12-01

    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. PMID:21198036

  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. 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

  10. 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

  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. High-throughput total cupric ion reducing antioxidant capacity of biological samples determined using flow injection analysis and microplate-based methods.

    PubMed

    Ribeiro, Joana P N; Magalhães, Luís M; Reis, Salette; Lima, José L F C; Segundo, Marcela A

    2011-01-01

    High-throughput cupric ion reducing antioxidant capacity (CUPRAC) methods were developed for assessment of total antioxidant capacity (TAC) in urine and serum, based on reduction of Cu(II)-neocuproine complex to highly colored Cu(I)-neocuproine complex, measured spectrophotometrically at 450 nm. The reaction time was significantly reduced from 30 to 4 min by application of a calibration compound (uric acid) with kinetic behavior similar to that shown by urine samples. The method was implemented in a microformat (96 well plates) and also in an automatic fashion (flow injection analysis, FIA). A determination throughput value of 288 h(-1) (microplate method) or of 15 h(-1) (automatic FIA) was attained. Application of both methods to human serum (SRM 909b, level I) and urines (n = 9) provided TAC values in agreement with those of the end-point batch method.

  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.

    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%.

  15. 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%.

  16. Synthesis of gold nano- and microplates in hexagonal liquid crystals.

    PubMed

    Wang, Luyan; Chen, Xiao; Zhan, Jie; Chai, Yongcun; Yang, Chunjie; Xu, Limei; Zhuang, Wenchang; Jing, Bo

    2005-03-01

    Single-crystalline gold nano- and microplates with triangular or hexagonal shapes are synthesized by reduction of HAuCl(4) in lyotropic liquid crystal (LLC) mainly made of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) block copolymers and water after adding a small amount of capping agents, cetyltrimethylammonium bromide (CTAB) or tetrabutylammonium bromide (TBAB). During the growth of such plates, capping agents play the crucial role. It is found that there is an optimal value of CTAB or TBAB concentration for producing microplates. The selective adsorption of CTAB or TBAB on certain crystallographic facets may be the key point of the supposed mechanism. Although LLC does not really act as a template, it provides an ordered structure confining CTAB as well as the nascent metal nuclei, which enhances the oriented attachment of nuclei and thus the consequent growth of single-crystal plates.

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

    PubMed

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

    2012-01-01

    Pressure differentials are routinely used to actuate flow in capillaries. We advance here an alternative means of flow generation that capitalizes on the extension of a liquid bridge achieved by the drawing of a rod through the action of surface tension. This meets the exigencies of creating controllable flow using simpler and more compact means. We found the ability to generate controllable flow to be strongly affected by the liquid bridge sustaining features, and that the use of rod diameters larger than the capillary was more conducive. The extensional flow resulting from the rupture of the liquid bridge was also found to have a strong circulation component which facilitated mixing. The approach here is highly amenable for use in capillary well microplates which have significant advantages over standard microplates. The features of this approach offer usage possibilities in biochemical applications in the field, such as in the leukocyte cell adhesion and hemagglutination tests of blood samples. PMID:21986403

  18. 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.

  19. 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.

  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. 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.

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

    PubMed

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

    2005-06-01

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

  3. A sensitive and quick microplate method to determine the minimal inhibitory concentration of plant extracts for bacteria.

    PubMed

    Eloff, J N

    1998-12-01

    Agar diffusion techniques are used widely to assay plant extracts for antimicrobial activity, but there are problems associated with this technique. A micro-dilution technique was developed using 96-well microplates and tetrazolium salts to indicate bacterial growth. p-Iodonitrotetrazolium violet [0.2 mg/ml] gave better results than tetrazolium red or thiazolyl blue. The method is quick, worked well with Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia coli and with non-aqueous extracts from many different plants. The method gave reproducible results; required only 10-25 microliters of extract to determine minimal inhibitory concentrations, distinguished between microcidal and microstatic effects, and provided a permanent record of the results. Using S. aureus, and a Combretum molle extract, the technique was 32 times more sensitive than agar diffusion techniques and was not sensitive to culture age of the test organism up to 24 hours. The S. aureus culture could be stored up to 10 days in a cold room with little effect on the assay results. This method was useful in screening plants for antimicrobial activity and for the bioassay-guided isolation of antimicrobial compounds from plants. MIC values determined for sulfisoxazole, norfloxacin, gentamicin, and nitrofuratoin were similar to values indicated in the literature but values obtained with trimethroprim and ampicillin were higher with some bacteria. PMID:9933989

  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. Evaporative preconcentration of fluorescent protein samples in capillary based microplates.

    PubMed

    Shao, Fenfen; Ng, Tuck Wah; Lye, Jonathan Kok Keung; Liew, Oi Wah

    2011-09-01

    The preconcentration of analytes is important in biochemical analysis as it offers the ability to detect for trace species, and increase signal-to-noise ratios when using optical sensing on fluorophores. A strong advantage of the evaporation technique lies in its ability to operate without the need of any energy source; albeit major challenges exist on how to increase the surface area exposure to air for heightened evaporation, ensure no further increases once specified analyte concentrations have been achieved, and not needing any intervening membranes. We demonstrate here that the droplet creation and retraction approach in capillary based microplates offers such abilities whilst at the same time facilitating mixing. PMID:21559858

  6. a Model of the Kinematics of the Danakil Microplate

    NASA Astrophysics Data System (ADS)

    Gloaguen, R.; Eagles, G.

    2001-12-01

    We present a regional approach to the tectonics of the Afar depression, in which we attempt to relate reconnaissance-scale data to overall movement of a Danakil microplate. Topographic, satellite, airphoto, magnetic and gravity data from the flanks and interior of the Afar depression, NE Africa, are used to assess differing models of the dispersal of the Danakil microplate from its western to its eastern margin. The data are compared to small circles about published instantaneous poles to find evidence of strike-slip and transverse features. A good fit to such features is made using a statistically-significant modern pole to the NNE of the Afar depression. A possible reconstruction about this pole, which reunites branches of an inherited shear zone, is presented. This reconstruction is not consistent with a `crank-arm' or `saloon-doors' mode of opening for the Afar depression. Instead we suggest that only recently has the Danakil microplate come into existence, after the westwards propagation of the Gulf of Aden rift into the Afar depression. Prior to this time movement of Danakil occurred very slowly about the same pole, but as part of the Somalian plate; as shown by the continuity of ancient flowline features onto the Nubian margin of the Main Ethiopian Rift. At 7 Ma, possibly due to the onset of oceanic-type accretion in Afar where the lithosphere was hot and thin already from the presence of a hotspot, Danakil accelerated eastwards about this pole. In response the Gulf of Aden rift propagated westwards to form a new plate boundary between Danakil and Somalia. The newly-formed Danakil microplate is thus an intermediate stage in the transfer of all or part of the Danakil horst to the Arabian plate from the Somalian plate. Further south, at the Main Ethiopian Rift, continental lithosphere did not rupture but the presence of focussed accretion at Afar in a hitherto-diffuse plate boundary precipitated the southward migration of the Somalia-Nubia pole to its present

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

    NASA Astrophysics Data System (ADS)

    Korja, Annakaisa

    2013-04-01

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

  8. Rapid 96-well plates DNA extraction and sequencing procedures to identify genome-wide transposon insertion sites in a difficult to lyse bacterium: Lactobacillus casei.

    PubMed

    Scornec, Hélène; Tichit, Magali; Bouchier, Christiane; Pédron, Thierry; Cavin, Jean-François; Sansonetti, Philippe J; Licandro-Seraut, Hélène

    2014-11-01

    Random transposon mutagenesis followed by adequate screening methods is an unavoidable procedure to characterize genetics of bacterial adaptation to environmental changes. We have recently constructed a mutant library of Lactobacillus casei and we aimed to fully annotate it. However, we have observed that, for L. casei which is a difficult to lyse bacterium, methods used to identify the transposon insertion site in a few mutants (transposon rescue by restriction and recircularization or PCR-based methods) were not transposable for a larger number because they are too time-consuming and sometimes not reliable. Here, we describe a method for large-scale and reliable identification of transposon insertion sites in a L. casei mutant library of 9250 mutants. DNA extraction procedure based on silica membranes in 96-column format was optimized to obtain genomic DNA from a large number of mutants. Then reliable direct genomic sequencing was improved to fit the obtained genomic DNA extracts. Using this procedure, readable and identifiable sequences were obtained for 87% of the L. casei mutants. This method extends the applications of a library of this type, reduces the number of insertions needed to be screened, and allows selection of specific mutants from an arrayed and stored mutant library. This method is applicable to any already existing mutant library (obtained by transposon or insertional mutagenesis) and could be useful for other bacterial species, especially for highly lysis-resistant bacteria species such as lactic acid bacteria.

  9. Detection of dengue viral RNA by microplate hybridization.

    PubMed

    Ruiz, B H; Zamora, M P; Liu, S

    1995-08-01

    Dengue virus infection is a major public health problem throughout tropical countries. In endemic areas, dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS) are common complications resulting in death. However, serological confirmation of dengue-related illness is often complicated and time-consuming. Detection of dengue viruses in clinical or field samples usually depends on virus isolation in susceptible cell lines or in mosquitoes, followed by viral protein identification using polyclonal or monoclonal antibodies. The increasing incidence of dengue virus infections has prompted increased efforts to develop rapid and reliable diagnostic techniques. A simple microplate hybridization method was developed for identification of viral RNA. Microplate hybridization is simpler than enzyme-linked immunosorbent assay and has several advantages over the conventional dot-blot hybridization method: (1) radioisotopes are not necessary; (2) synthetic oligonucleotide for the probe is not needed; (3) the time required for washing of the solid phase is greatly reduced; and (4) baking is eliminated. The results show that this procedure is sensitive, rapid and easy to perform.

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

    USGS Publications Warehouse

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

    2003-01-01

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

  11. Microplate Technique for Determining Accumulation of Metals by Algae

    PubMed Central

    Hassett, James M.; Jennett, J. Charles; Smith, James E.

    1981-01-01

    A microplate technique was developed to determine the conditions under which pure cultures of algae removed heavy metals from aqueous solutions. Variables investigated included algal species and strain, culture age (11 and 44 days), metal (mercury, lead, cadmium, and zinc), pH, effects of different buffer solutions, and time of exposure. Plastic, U-bottomed microtiter plates were used in conjunction with heavy metal radionuclides to determine concentration factors for metal-alga combinations. The technique developed was rapid, statistically reliable, and economical of materials and cells. Results (expressed as concentration factors) were in reasonably good agreement with literature values. All species of algae studied removed mercury from solution. Green algae proved better at accumulating cadmium than did blue-green algae. No alga studied removed zinc, perhaps because cells were maintained in the dark during the labeling period. Chlamydomonas sp. proved superior in ability to remove lead from solution. PMID:16345764

  12. Photosensitizer Adhered to Cell Culture Microplates Induces Phototoxicity in Carcinoma Cells

    PubMed Central

    Ziegler, Verena; Kiesslich, Tobias; Krammer, Barbara; Plaetzer, Kristjan

    2013-01-01

    In vitro experiments in plastic receptacles are the basis of characterization of new photosensitizers (PSs) for the photodynamic therapy. We recently reported that lipophilic PSs adhere to cell culture microplates in a kinetic-like manner (Engelhardt et al., 2011). In the current study, we examined the interaction and phototoxic effects of the microplate-adhered PS in cancer cells. Therefore, we preloaded microplates with hypericin, Foscan, PVP-hypericin, or aluminum (III) phthalocyanine tetrasulfonate chloride (AlPCS4) for 24 hours and measured the PS distribution after addition of A431 human carcinoma cells: following another 24 hours up to 68% of hypericin were detected in the cell fraction. The hydrophilic PVP-hypericin and AlPCS4 also diffused into the cells, but the quantities of PS adherence were considerably lower. Microplate-adhered Foscan appeared not to be redistributed. In contrast to the hydrophilic PSs, the cellular phototoxicity of microplate-adhered lipophilic PS was high, independent of whether the PS (i) was pre-loaded onto microplates or (ii) added simultaneously with the cells or (iii) one day after cell seeding. Based on these results, we suggest testing lipophilic PS dyes for their adherence to microplates. Furthermore, the ability of plastic materials to (reversibly) store PSs might represent a new approach for the PS delivery or the development of antimicrobial coatings. PMID:23509741

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

    PubMed

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

    2002-01-01

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

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

    PubMed

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

    2006-02-01

    A 96-well single-pot protein precipitation, liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the determination of muraglitazar, a PPAR alpha/gamma dual agonist, in human plasma. The internal standard, a chemical analogue, was dissolved in acetonitrile containing 0.1% formic acid. The solvent system was also served as a protein precipitation reagent. Human plasma samples (0.1 mL) and the internal standard solution (0.3 mL) were added to a 96-well plate. The plate was vortexed for 1 min and centrifuged for 5 min. Then the supernatant layers were directly injected into the LC/MS/MS system. The chromatographic separation was achieved isocratically on a Phenomenox C18(2) Luna column (2 mm x 50 mm, 5 microm). The mobile phase contained 20/80 (v/v) of water and acetonitrile containing 0.1% formic acid. Detection was by positive ion electrospray tandem mass spectrometry on a Sciex API 3000. The standard curve, which ranged from 1 to 1000 ng/mL, was fitted to a 1/x weighted quadratic regression model. This single-pot approach effectively eliminated three time consuming sample preparation steps: sample transfer, dry-down, and reconstitution before the injection, while it preserved all the benefits of the traditional protein precipitation. By properly adjusting the autosampler needle offset level, only the supernatant was injected, without disturbing the precipitated proteins in the bottom. As a result, the quality of chromatography and column life were not compromised. After more than 600 injections, there was only slightly increase of column back-pressure. The validation results demonstrated that this method was rugged and provide satisfactory precision and accuracy. The method has been successfully applied to analyze human plasma samples in support of a first-in-man study. This method has also been validated in monkey and mouse plasma for the determination of muraglitazar. PMID:16388995

  15. 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

  16. 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.

  17. 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

  18. 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

  19. 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.

  20. 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.

  1. 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

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

    PubMed

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

    2016-08-19

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

  3. 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.

  4. Direct monitoring of molecular recognition processes using fluorescence enhancement at colloid-coated microplates.

    PubMed

    Lobmaier, C; Hawa, G; Götzinger, M; Wirth, M; Pittner, F; Gabor, F

    2001-01-01

    Direct monitoring of recognition processes at the molecular level is a valuable tool for studying reaction kinetics to assess affinity constants (e.g. drugs to receptors) and for designing rapid single step immunoassays. Methods currently used to gain information about binding processes predominantly depend on surface plasmon resonance. These systems use excitation with coherent light in attenuated total reflection geometry to obtain discrimination between surface-bound and free molecules in solution. Therefore labeling of the compounds is not necessary, but due to the complexity of the measuring setup the method is rather costly. In this contribution we present a simple method for performing kinetic single step biorecognition assays with fluorophore labeled compounds using the fluorescence enhancement properties of surface bound silver colloids. Silver colloids are bound to standard microplates via silanization of the plastic surface. Fluorophores close to this colloid coated surface show a significant gain in fluorescence compared to fluorophores farther away in the bulk solution. Therefore discrimination between surface bound and free fluorophores is possible and the binding of, for example, fluorophore labeled antibodies to antigens immobilized on the colloid surface results in increasing fluorescence intensity. Utilization of standard microplates makes this method fully compatible with conventional microplate processing and reading devices. Neither excitation with coherent laser light nor ATR geometry is required, the measurement is performed in a standard fluorescence microplate reader in front face geometry with a xenon flash lamp as excitation source. Methods for the preparation of colloid-coated microplates and fluorescence-enhanced biorecognition assays are presented. Additionally the dependence of the system performance on the structure and properties of the metal colloid coated surface is described. A two-component biorecognition model system shows a

  5. High-throughput method for a kinetics analysis of the high-pressure inactivation of microorganisms using microplates.

    PubMed

    Hasegawa, Toshimi; Hayashi, Manabu; Nomura, Kazuki; Hayashi, Mayumi; Kido, Miyuki; Ohmori, Tsuneo; Fukuda, Masao; Iguchi, Akinori; Ueno, Shigeaki; Shigematsu, Toru; Hirayama, Masao; Fujii, Tomoyuki

    2012-06-01

    Using microplates as pressure and cultivation vessels, a high-throughput method was developed for analyzing the high-pressure inactivation kinetics of microorganisms. The loss of viability from a high-pressure treatment, measured based on the growth delay during microplate cultivation, showed reproducibility with the conventional agar plate method and was applicable for the kinetics analysis.

  6. 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

  7. 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.

  8. 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.

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

    PubMed

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

    2012-11-01

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

  10. A rapid method for the determination of perfluoroalkyl substances including structural isomers of perfluorooctane sulfonic acid in human serum using 96-well plates and column-switching ultra-high performance liquid chromatography tandem mass spectrometry.

    PubMed

    Salihovic, Samira; Kärrman, Anna; Lindström, Gunilla; Lind, P Monica; Lind, Lars; van Bavel, Bert

    2013-08-30

    To facilitate high-throughput analysis suitable for large epidemiological studies we developed an automated column-switching ultra-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for determination of perfluorocarboxylic acids (PFCAs; C5, C6, C7, C8, C9, C10, C11, C12, and C13), perfluoroalkyl sulfonic acids (PFSAs; C4, C6, C8, and C10), perfluorooctane sulfonamide (PFOSA), and five groups of structural perfluorooctane sulfonic acid (PFOS) isomers in human serum or plasma. The analytical procedure involves rapid protein precipitation using 96-well plates followed by an automated sample clean-up using an on-line trap column removing many potentially interfering sample components while through the mobile phase gradient the target analytes are eluted onto the analytical column for further separation and subsequent mass detection. The method was linear (R(2)<0.995) at concentrations ranging from 0.01 to 60ngmL(-1) with method detection limits ranging between 0.01 and 0.17ngmL(-1) depending on the analyte. The developed method was precise, with repeatability (n=7) and reproducibility (n=103) coefficients of variation between 2% and 20% for most compounds including PFOS (2% and 8%) and its structural isomers (2-6% and 4-8%). The method was in conformity with a standard reference material. The column-switching HPLC-MS/MS method has been successfully applied for the determination of perfluoroalkyl substances including structural PFOS isomers in human plasma from an epidemiological study.

  11. 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.

  12. 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.

  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. 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.

  15. 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

  16. 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

  17. 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.

  18. 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

  19. 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

  20. Fabrication of surface plasmon resonators by nanoskiving single-crystalline gold microplates

    NASA Astrophysics Data System (ADS)

    Wiley, Benjamin J.; Lipomi, Darren J.; Bao, Jiming; Capasso, Federico; Whitesides, George M.

    2009-02-01

    This paper demonstrates the sectioning of chemically synthesized, single-crystalline microplates of gold with an ultramicrotome to produce single-crystalline nanowires. This method produces collinearly aligned nanostructures with small, regular changes in dimension with each consecutive cross-section. The diamond knife cuts cleanly through microplates 100 nm thick without bending the resulting nanowire, and cuts through the sharp edges of a crystal to generate nanoscale tips. This paper demonstrates that the smooth surface of the single-crystalline gold nanowires allows them to guide plasmons with lower loss than rough, polycrystalline nanowires, and that the sharp tips on the singlecrystalline nanowires serve as optical antenna that selectively couple light into the nanowire at the resonance frequency of the sharp tip.

  1. A fast optical method for the determination of liquid levels in microplates.

    PubMed

    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.

  2. 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

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

    PubMed

    Shintaku, Jonathan; Guttridge, Denis C

    2016-01-01

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

  4. Osteoplastic laminotomy using titanium microplates for reconstruction of the laminar roof: a technical note.

    PubMed

    Wiedemayer, H; Schoch, B; Stolke, D

    1998-01-01

    A technique to reconstruct the posterior elements of the spinal canal in osteoplastic laminotomy is described. The use of titanium microplates allows a stable fixation of the laminar arch with optimal realignment of the lamina cut ends. The titanium implants are fully compatible with magnetic resonance imaging. The author's experience with thirty patients is reported. The method proved to be simple and effective with distinct advantages over conventional techniques. PMID:9795940

  5. 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

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

    PubMed

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

    2007-05-01

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

  7. Size-dependent electromechanical coupling behaviors of circular micro-plate due to flexoelectricity

    NASA Astrophysics Data System (ADS)

    Li, Anqing; Zhou, Shenjie; Qi, Lu

    2016-10-01

    In this paper, the flexoelectric theory is re-expressed by a set of orthogonal components of strain gradient tensor. The general formulations of flexoelectric theory in orthogonal curvilinear coordinates are derived and, then, are specified for the case of cylindrical coordinates. A flexoelectric circular micro-plate model is established based on the current formulations in cylindrical coordinates to evaluate its size-dependent static and dynamic responses. The governing equations, boundary conditions and initial conditions are obtained according to the Hamilton's principle. A static bending problem of simply supported axisymmetric circular micro-plate is solved in two cases, of which one is subjected to a distributed load and the other is subjected to a voltage across the plate thickness. And the free vibration problem of a simply supported circular micro-plate is also analyzed. The bending numerical results show that both the deflection and the electric potential exhibit obvious size dependency in the two cases. Both the induced electric potential in direct flexoelectric effect and the induced deflection in inverse flexoelectric effect decrease as the decrease in flexoelectric coefficient and even disappear when the flexoelectric coefficient equals zero. Moreover, the numerical results of free vibration demonstrate the dimensionless natural frequency shows obvious size effect, while the influence of flexoelectric coefficient on dimensionless natural frequency is negligible.

  8. 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.

  9. Thermal and rheological properties of the NW sector of the Adria microplate between Alps and Apennines (Northern Italy)

    NASA Astrophysics Data System (ADS)

    Vigano', Alfio; Della Vedova, Bruno; Ranalli, Giorgio; Martin, Silvana; Scafidi, Davide

    2010-05-01

    The present structural setting of the NW sector of the Adria microplate, overridden by the advancing fronts of the Alpine and Apennine chains from nearly opposite directions, is the result of polyphase tectonic deformation beginning in the Late Cretaceous. The deformation was accomodated at different times by S-verging (Alpine front) and N-verging (Appennine front) thrust systems, in which fault patterns are strongly affected by inherited Mesozoic extensional N-S oriented faults. We study the thermal and rheological properties of the central part of the present Po Plain (approximately 44.5-45.7 ° N lat, 9.4-11.2 ° E long), which records the convergence of the Alpine and Apennine orogenic fronts. The present thermal regime of the crust is constrained by geological and geophysical results from oil exploration. A set of 38 deep boreholes (Eni Exploration & Production) provides lithology and temperature data (bottom hole, drill stem, and production test temperatures) down to 6-7 km of depth. Bottom hole data were processed to estimate undisturbed formation temperatures. The thermal conductivity was estimated from lithology and logging data. The thermal resistance method was applied to verify the appropriateness of purely conductive and steady-state heat transfer conditions. Temperature-depth plots show two clearly distinguishable average geotherms, corresponding to the Western (W) and Eastern (E) areas of the NW Adria microplate sector. The two zones show significantly different crustal structures. The internal consistency of data in each zone and the difference between the two groups of data, which is larger than measurement uncertainties, confirm the validity of spatial zonation as a first-order working hypothesis. Temperature measurements in the upper ~7 km of the crust are used to constrain 1D thermal models at the crustal scale. Although the Adria microplate in this area is expected to exhibit lateral heterogeneities due to its complex 3D structure, we derive

  10. The Setting of the Okhotsk-Sea Micro-Plate Northern Boundary and its Seismicity

    NASA Astrophysics Data System (ADS)

    Sedov, B. M.

    2004-12-01

    The Sea of Okhotsk is surrounded by seismic zones. That is why we believe that its water area corresponds spatially to hard platform. The Okhotsk-sea micro-plate in triangular form is located at the joint of the Pacific, North-Amerikan, and Eurasian plates. It is bounded from the west by sub-meridional zone of deep sesmogenic faults spreading through the Sakhalin island. From the east, micro-plate is bounded by subduction zones, extending along the Kuril Islans and the eastern coast of Kamchatka. The northern boundary of the Okhotsk-sea plate is considered differently by different authors. The analysis of data of the marine seismic exploration by the method of common deep point, deep seismic sounding, gravitation field and the anomalous magnetic field, and also the seismological observation by the equipment make possible to conclude that the northern boundary of the Okhotsk-sea micro-plate has the sublatitudinal extending and spatially coincides with wide zone of deep faults confided to the coast. On land and offshore, this zone is marked by the line of riftogenic basins filled with Cenozoic continental sediments. On the continent, their thickness is not more than 1500-2000m, although on the offshore it reaches 8-10 km. The basins are mostly spread in meridian direction. The epicenters of the crust earthquakes are spatially related with these basins. This fact testifies of the right-lateral faults. Cenozoic sediments in basins of land are broken by numerous fault-shifts with the summary amplitude of horizontal displacements up to 1.5 km and by the vertical ones up to the first hundreds of meters. The northern boundary of the Okhotsk-sea micro-plate is marked by the change of the Earth's crust type: the continental type on the north, and transitional type on the offshore. Geological and geophisical data make possible to suppose that the northern boundary of micro-plate in primagadanskaya part coincides with the Benioff paleozone that closed its active existence in

  11. Enterococcus Faecalis Biofilm. Formation and Development in Vitro Observed by Scanning Electron Microscopy.

    PubMed

    Bulacio, María de Los Á; Galván, Lucas R; Gaudioso, Cristina; Cangemi, Rosa; Erimbaue, Marta I

    2015-12-01

    Biofilm produced by Enterococcus faecalis isolated from root canals was detected by growing it on microplates and using 10% crystal violet stain, elution with alcohol and three procedures: no fixation, heat fixation and 10% formaldehyde fixation. The biofilm was evaluated using a Versamax Microplate Reader (USA). Twenty sterile root portions were incubated in TS broth with E. faecalis (108) for 48 hours, 4, 7, 14 and 30 days, after which they were processed and observed by scanning electron microscopy (SEM). Significantly more biofilm was found on the microplates for formaldehyde fixation than for heat fixation or no fixation (ANOVA p<0.0001). SEM showed E. faecalis growth at all times and biofilm development as from 14 days' incubation. Fixation with 10% formaldehyde was the most appropriate technique for detecting E. faecalis biofilm development on microplates. SEM confirmed biofilm formation after 14 days incubation.

  12. 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.

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

    PubMed

    Li, Pu; Fang, Yuming

    2015-03-25

    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.

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

    PubMed Central

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

    2016-01-01

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

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

    DOE PAGES

    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

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

    NASA Astrophysics Data System (ADS)

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

    2016-04-01

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

  19. 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

  20. 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

  1. Rapid and reproducible infectivity end-point titration of virulent phage in a microplate system.

    PubMed

    Fridholm, Helena; Everitt, Einar

    2005-09-01

    The standard method for measuring the number of infectious phages in solution has traditionally been the plaque forming assay. An alternative method is described where the number of lytic, infectious phages is determined in an endpoint titration assay adapted for a microplate system. In this model system, susceptible Escherichia coli B6 at a density of 4 x 10(7) cells/ml, were mixed with an equal volume (100 microl) of PhiX174 diluted serially in a microtest plate. After 3h of incubation on a microplate shaker the endpoint was determined spectrophotometrically and calculated according to the method of Reed and Muench. A well was considered positive for infection if the OD630-value was < or = 10% compared to the OD630-value of the negative control of uninfected cells. ID50-titers were 2.5x higher than the PFU-titers (CV 15%) and the intra assay reproducibility revealed a CV of 9%. The method has several advantages as compared with the conventional PFU-titration. It is less time and material consuming with the possibility to assess several samples at the same time.

  2. 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

  3. 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.

  4. [Metabolic properties of the microbial community in the biofilters using biolog microplates].

    PubMed

    Xi, Jin-Ying; Hu, Hong-Ying; Jiang, Jian; Qian, Yi

    2005-07-01

    It is very important to know the structure and metabolic function of the microbial community in a bioreactor in order to improve its performance. In this study, two biofilters, packed with wood chips and granular activated carbons respectively, were operated for 160 days to treat toluene gas. The metabolic profiles of the microbial communities in the biofilters were monitored using Biolog microplates periodically during the experiments. The metabolic activities of the microorganisms in both biofilters were observed to decrease during long-term operation. According to the results of principle components analysis, the metabolic profiles of the microbial communities did not change much in the former period of the operation, but they changed in the inlet layers on day 103 and changed throughout the filter beds on day 160. The variation of the metabolic profiles in both biofilters showed little difference, which suggested that the packing media had little effect on them during long-term operation. Among the 95 carbon sources in Biolog microplate, carboxylic acids and amino acids were much easier to be utilized by the microorganisms in the biofilters than the other carbon

  5. 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

  6. 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

  7. 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.

  8. 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

  9. 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

  10. 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

    2016-08-05

    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.

  11. 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

  12. 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

  13. 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...

  14. 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...

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

    NASA Astrophysics Data System (ADS)

    Farokhi, Hamed; Ghayesh, Mergen H.

    2016-06-01

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

  16. 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.

  17. 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

  18. Analysis and interpretation of microplate-based oxygen consumption and pH data.

    PubMed

    Divakaruni, Ajit S; Paradyse, Alexander; Ferrick, David A; Murphy, Anne N; Jastroch, Martin

    2014-01-01

    Breakthrough technologies to measure cellular oxygen consumption and proton efflux are reigniting the study of cellular energetics by increasing the scope and pace with which discoveries are made. As we learn the variation in metabolism between cell types is large, it is helpful to continually provide additional perspectives and update our roadmap for data interpretation. In that spirit, this chapter provides the following for those conducting microplate-based oxygen consumption experiments: (i) a description of the standard parameters for measuring respiration in intact cells, (ii) a framework for data analysis and normalization, and (iii) examples of measuring respiration in permeabilized cells to follow up results observed with intact cells. Additionally, rate-based measurements of extracellular pH are increasingly used as a qualitative indicator of glycolytic flux. As a resource to help interpret these measurements, this chapter also provides a detailed accounting of proton production during glucose oxidation in the context of plate-based assays. PMID:25416364

  19. The Adriatic region: An independent microplate within the Africa-Eurasia collision zone

    NASA Astrophysics Data System (ADS)

    Battaglia, Maurizio; Murray, Mark H.; Serpelloni, Enrico; Bürgmann, Roland

    2004-05-01

    We use GPS measurements and block modeling to investigate the present-day deformation of the Adriatic region, whose kinematics within the Nubia-Eurasia plate boundary zone is not well constrained and remains controversial. Block modeling allows us to compute rigid-plate angular velocities while accounting for elastic strain accumulation along block-bounding faults. Results suggest that the Adriatic is a microplate (Adria) and that the southern boundary with the Nubia plate and the Aegean domain may be located along the Apulia Escarpment and the Kefallinia fault. Geodetic data alone cannot discriminate between a single block (AP) or a two blocks (GDAP) description of Adria, but the GDAP model predicts boundary slip rates that are in better agreement with observations from previous studies.

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

    PubMed Central

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

    1988-01-01

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

  1. Analysis and interpretation of microplate-based oxygen consumption and pH data.

    PubMed

    Divakaruni, Ajit S; Paradyse, Alexander; Ferrick, David A; Murphy, Anne N; Jastroch, Martin

    2014-01-01

    Breakthrough technologies to measure cellular oxygen consumption and proton efflux are reigniting the study of cellular energetics by increasing the scope and pace with which discoveries are made. As we learn the variation in metabolism between cell types is large, it is helpful to continually provide additional perspectives and update our roadmap for data interpretation. In that spirit, this chapter provides the following for those conducting microplate-based oxygen consumption experiments: (i) a description of the standard parameters for measuring respiration in intact cells, (ii) a framework for data analysis and normalization, and (iii) examples of measuring respiration in permeabilized cells to follow up results observed with intact cells. Additionally, rate-based measurements of extracellular pH are increasingly used as a qualitative indicator of glycolytic flux. As a resource to help interpret these measurements, this chapter also provides a detailed accounting of proton production during glucose oxidation in the context of plate-based assays.

  2. Typing dinucleotide repeat loci using microplate array diagonal gel electrophoresis: proof of principle.

    PubMed

    Rodríguez, Santiago; Chen, Xiao-He; Day, Ian N M

    2004-04-01

    Polymorphic dinucleotide repeat loci ('microsatellite markers') are found in varying abundance throughout the genomes of most organisms. They have been extensively used for genetic studies, but conventional techniques used for their genotyping require sophisticated equipment. Microplate array diagonal gel electrophoresis (MADGE) has previously been extended to economical high-throughput genotyping of trinucleotide and tetranucleotide microsatellite amplicons. However, the capability of this technique to resolve the alleles of dinucleotide repeat loci has not been explored previously. Here we show that a modified microsatellite-MADGE approach can provide sufficient resolution for dinucleotide repeat typing. This enables economical and convenient set up for analysis of single markers in many samples in parallel, suitable, for example, for population association studies.

  3. [Evaluation of DNA-DNA hybridization method for identification of mycobacteria using a colorimetric microplate kit].

    PubMed

    Yamazaki, T; Takahashi, H; Nakamura, R M

    1993-01-01

    DNA-DNA hybridization was applied for identification of mycobacteria and developed as a kit "microplate hybridization kit" (refers to MPHD) by Kobayashi Pharmaceutical Co. We received test samples of the microplates from the company and examined them for their and reliability using 180 mycobacterial strains of 21 species kept in our laboratory. The results of identification by MPHD were 100% identical to those of biochemical identification in the type or reference strains of mycobacteria, showing good reliability of MPHD method. Among clinical isolates, there were six M. tuberculosis strains which did not show typical characteristics for M. tuberculosis, i.e., niacin test negative or nitrate reduction weak positive, but all of these were identified as M. tuberculosis complex by MPHD method. Some strains from clinical isolates showed difference in identification between MPHD and biochemical methods: M. avium complex, identified biochemically were divided into M. avium and M. intracellulare by MPHD, M. fortuitum complex by biochemical identification were distinguished as M. fortuitum and M. chelonae by MPHD. Further, M. chelonae were separated into M. chelonae subsp. chelonae and M. chelonae subsp. abscessus by MPHD. M. peregrinum has been considered as a synonym of M. fortuitum, but we could distinguish M. peregrinum from M. fortuitum clearly by MPHD method. Thus, it is suggested that M. peregrinum and M. fortuitum are different species. Keys for getting reliable results using the MPHD kit are: (1) appropriate amount of bacteria for use, (2) purification of DNA, (3) enough deproteinization, and (4) appropriate timing to read colorimetry measurement of the plate. PMID:8437424

  4. 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.

  5. 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

  6. The Encapsidated Genome of Microplitis demolitor Bracovirus Integrates into the Host Pseudoplusia includens ▿ ‡

    PubMed Central

    Beck, Markus H.; Zhang, Shu; Bitra, Kavita; Burke, Gaelen R.; Strand, Michael R.

    2011-01-01

    Polydnaviruses (PDVs) are symbionts of parasitoid wasps that function as gene delivery vehicles in the insects (hosts) that the wasps parasitize. PDVs persist in wasps as integrated proviruses but are packaged as circularized and segmented double-stranded DNAs into the virions that wasps inject into hosts. In contrast, little is known about how PDV genomic DNAs persist in host cells. Microplitis demolitor carries Microplitis demolitor bracovirus (MdBV) and parasitizes the host Pseudoplusia includens. MdBV infects primarily host hemocytes and also infects a hemocyte-derived cell line from P. includens called CiE1 cells. Here we report that all 15 genomic segments of the MdBV encapsidated genome exhibited long-term persistence in CiE1 cells. Most MdBV genes expressed in hemocytes were persistently expressed in CiE1 cells, including members of the glc gene family whose products transformed CiE1 cells into a suspension culture. PCR-based integration assays combined with cloning and sequencing of host-virus junctions confirmed that genomic segments J and C persisted in CiE1 cells by integration. These genomic DNAs also rapidly integrated into parasitized P. includens. Sequence analysis of wasp-viral junction clones showed that the integration of proviral segments in M. demolitor was associated with a wasp excision/integration motif (WIM) known from other bracoviruses. However, integration into host cells occurred in association with a previously unknown domain that we named the host integration motif (HIM). The presence of HIMs in most MdBV genomic DNAs suggests that the integration of each genomic segment into host cells occurs through a shared mechanism. PMID:21880747

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

    NASA Astrophysics Data System (ADS)

    Schmid, Stefan

    2014-05-01

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

  8. Epstein-barr virus VCA IgM and EBNA IgG antibodies titered by immunofluorescence in microplates. A semi-automated method based on microtiter system.

    PubMed

    Lamy, M E; Favart, A M; Leclercq, M F; Segas, M; Mendez, M; Lhoir, C; Cornu, C; Burtonboy, G

    1982-01-01

    We describe methods of immunofluorescence in microplates for titration of EBV VCA-IgM and EBNA-IgG antibodies and compare the sensitivity of the methods with glass slides and with microplates, showing the reproducibility of the methods. Results of VCA-IgM and -IgG, EA and EBNA titers obtained in four groups of patients are given, comprising cases of infectious mononucleosis, renal graft recipients, lymphoproliferative diseases and controls.

  9. 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.

  10. 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

  11. Assessing the Rigidity of the Puerto Rico-Virgin Islands Microplate: Results from GPS Geodesy in the British Virgin Islands

    NASA Astrophysics Data System (ADS)

    Stamps, D. S.; Baldwin, A.; Turner, H.; Mattioli, G.; Jansma, P.

    2005-12-01

    Recent studies provide evidence of three microplates defined by the Puerto Rico trench to the north and the Muertos trough to the south in the boundary zone between the North America and Caribbean plates in the northeastern Caribbean. From west to east, these are the Gonave, Hispaniola, and Puerto Rico-northern Virgin Islands (PRVI) microplates. The eastern and western boundaries of the Gonave and Hispaniola microplates and the western boundary of the PRVI microplate have been defined in previous studies. However, the easternmost terminus of PRVI is undetermined, but suspected to lie either within or at the eastern terminus of the British Virgin Islands (BVI). Previously reported geodetic data indicates east-west extension across Puerto Rico and between eastern Puerto Rico and Virgin Gorda in the BVI. Our 2005 campaign focused on defining the easternmost terminus of PRVI and testing the extensional hypothesis by collecting surface motion data using Global Positioning System (GPS) geodesy. We collected data at sites on Tortola and Anegada in the BVI and concluded that the GPS-derived velocities of Tortola and Anegada with respect to the Caribbean are 5.71 + 5.5 mm/yr to the NNW (one sigma) and 3.12 + 2.7 mm/yr to the WNW (one sigma), respectively. These velocities are similar to those of eastern Puerto Rico. No motion relative to the Caribbean for Tortola and Anegada also is a possibility within error. If the GPS-derived velocities of Tortola and Anegada are 3-5 mm/yr, little to no east-west extension occurs between the BVI and eastern Puerto Rico, implying that PRVI is rigid at the mm/yr level and that the eastern terminus of PRVI coincides with the eastern end of the BVI near Virgin Gorda.

  12. Biomineralized anisotropic gold microplate-macrophage interactions reveal frustrated phagocytosis-like phenomenon: a novel paclitaxel drug delivery vehicle.

    PubMed

    Singh, Ajay Vikram; Batuwangala, Madu; Mundra, Ruchir; Mehta, Krunal; Patke, Sanket; Falletta, Ermelinda; Patil, Rajendra; Gade, W N

    2014-08-27

    This study reports a facile biomineralization route for gold microplates (GMPs) synthesis using bovine serum albumin (BSA) as a reductant and stabilizing agent. Adding BSA to HAuCl4 solution yields spontaneous versatile anisotropic and partially hollow GMPs upon aging. We hypothesize that the instantaneous protein denaturation at low pH enabled access to serine and threonine hydroxyl, and sulfhydryl groups of BSA, which act as a reductant and stabilizer, respectively. This reaction could be hastened by increasing the temperature well beyond 65 °C. Transmission electron microscopy/X-ray diffraction studies revealed highly crystalline and anisotropic structures (triangle, pentagon, and rectangle). Atomic force microscopy/scanning electron microscopy analyses demonstrated unique morphology of microplates with a partially void core and BSA mineralized edge structure. RAW 264.7 mice peritoneal macrophage-microplate interaction studies using live cell confocal imaging reveal that cells are capable of selectively internalizing smaller GMPs. Large GMPs are preferentially picked with sharp vertices but cannot be internalized and exhibit frustrated phagocytosis-like phenomenon. We explored particle phagocytosis as an actin mediated process that recruits phagosome-like acidic organelles, shown by a lysosensor probe technique. The biocompatible GMPs exhibited ∼70% paclitaxel (PCL) loading and sustained release of PCL, showing antitumor activity with the MCF-7 cell line, and could be a novel drug carrier for breast cancer therapy.

  13. Development, optimization and validation of a rapid colorimetric microplate bioassay for neomycin sulfate in pharmaceutical drug products.

    PubMed

    Francisco, Fabiane Lacerda; Saviano, Alessandro Morais; Pinto, Terezinha de Jesus Andreoli; Lourenço, Felipe Rebello

    2014-08-01

    Microbiological assays have been used to evaluate antimicrobial activity since the discovery of the first antibiotics. Despite their limitations, microbiological assays are widely employed to determine antibiotic potency of pharmaceutical dosage forms, since they provide a measure of biological activity. The aim of this work is to develop, optimize and validate a rapid colorimetric microplate bioassay for the potency of neomycin in pharmaceutical drug products. Factorial and response surface methodologies were used in the development and optimization of the choice of microorganism, culture medium composition, amount of inoculum, triphenyltetrazolium chloride (TTC) concentration and neomycin concentration. The optimized bioassay method was validated by the assessment of linearity (range 3.0 to 5.0μg/mL, r=0.998 and 0.994 for standard and sample curves, respectively), precision (relative standard deviation (RSD) of 2.8% and 4.0 for repeatability and intermediate precision, respectively), accuracy (mean recovery=100.2%) and robustness. Statistical analysis showed equivalency between agar diffusion microbiological assay and rapid colorimetric microplate bioassay. In addition, microplate bioassay had advantages concerning the sensitivity of response, time of incubation, and amount of culture medium and solutions required.

  14. 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

  15. 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

  16. DNA based classification of food associated Enterobacteriaceae previously identified by Biolog GN Microplates.

    PubMed

    Olsson, Crister; Ahrné, Siv; Pettersson, Bertil; Molin, Göran

    2004-03-01

    Enterobacteriaceae are frequently isolated from food products and it is essential to have methods for correct identification for both food hygiene and epidemiology reasons. Phenotypic methods are not always sufficient and have to be supplemented by DNA based methods. In the present study, 70 strains of Enterobacteriaceae derived from milk, fish and meat that had previously been identified by Biolog GN Microplates were genomically classified together with 15 representative type strains of species of Enterobacteriaceae. The field strains were dominated by Hafnia alvei, Serratia liquefaciens and Rahnella aquatilis. All strains were subjected to temporal temperature gel electrophoresis (TTGE) analysis using amplicons encompassing the V3, V4 and V9 variable regions of the 16S rRNA gene. Selected strains were analysed by ribotyping and partial 16S rDNA sequencing. The type strains were differentiated into 10 different TTGE groups. Two of the groups contained two type strains. Enterobacter aerogenes and Klebsiella planticola were not distinguished due to their identical sequences and Yersinia ruckeri and Citrobacter freundii showed the same migration pattern. The 70 food strains could be differentiated into 14 TTGE groups where 33 strains (47.1%) could be assigned to TTGE groups including type or reference strains. Rahnella strains were dispersed into three TTGE groups of which one group corresponded to Rahnella genomospecies 1 and one to genomospecies 3. The grouping of Rahnella strains was supported by ribotyping and phylogenetic analysis. TTGE can be a useful additional tool for identification on the species level of food related Enterobacteriaceae.

  17. Viscous and acoustic losses in length-extensional microplate resonators in liquid media

    NASA Astrophysics Data System (ADS)

    Ruiz-Díez, Víctor; Hernando-García, Jorge; Manzaneque, Tomás; Kucera, Martin; Schmid, Ulrich; Sánchez-Rojas, José Luis

    2015-02-01

    Damping mechanisms in the length-extensional mode of rectangular, mid-point supported microplate resonators immersed in liquid are studied. Piezoelectrically excited structures with different lengths and thicknesses were designed, fabricated, and characterized both optically and electrically in isopropanol. The experimental quality factors were compared to the results of Finite Element Method (FEM) simulations and the two main mechanisms of losses, i.e., acoustic and viscous losses, were identified. Analytical models for those two mechanisms are presented and the effects of the geometry on the in-liquid performance of the resonators are discussed. By applying these models, we found that for a given thickness, a maximum quality factor is reached at a critical length, resulting from the balance between acoustic and viscous losses. To further increase quality factors, a quarter wavelength fluid cavity was implemented, thereby reducing acoustic losses; an increase over 40% in the quality factor was predicted by a 2D FEM model including the cavity, and a quality factor as high as 145 was measured for a 3 mm long and 93 μm thick resonator in this configuration.

  18. 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

  19. Modeling the liquid filling in capillary well microplates for analyte preconcentration.

    PubMed

    Yu, Yang; Wang, Xuewei; Ng, Tuck Wah

    2012-06-15

    An attractive advantage of the capillary well microplate approach is the ability to conduct evaporative analyte preconcentration. We advance the use of hydrophobic materials for the wells which apart from reducing material loss through wetting also affords self entry into the well when the droplet size reduces below a critical value. Using Surface Evolver simulation without gravity, we find the critical diameters D(c) fitting very well with theoretical results. When simulating the critical diameters D(c)(G) with gravity included, the gravitational effect could only be ignored when the liquid volumes were small (difference of 5.7% with 5 μL of liquid), but not when the liquid volumes were large (differences of more than 22% with 50 μL of liquid). From this, we developed a modifying equation from a series of simulation results made to describe the gravitational effect. This modifying equation fitted the simulation results well in our simulation range (100°≤θ≤135° and 1 μL≤V≤200 μL). In simulating the condition of multiple wells underneath each droplet, we found that having more holes did not alter the critical diameters significantly. Consequently, the modifying relation should also generally express the critical diameter for multiple wells under a droplet.

  20. 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.

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

    PubMed

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

    2016-07-01

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

  2. 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

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

    PubMed

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

    2015-11-01

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

  4. Odorant binding characteristics of three recombinant odorant binding proteins in Microplitis mediator (Hymenoptera: Braconidae).

    PubMed

    Li, Keming; Wang, Shanning; Zhang, Kang; Ren, Liyan; Ali, Abid; Zhang, Yongjun; Zhou, Jingjiang; Guo, Yuyuan

    2014-06-01

    Odorant binding proteins (OBPs) are believed to be important for transporting semiochemicals through the aqueous sensillar lymph to the olfactory receptor cells within the insect antennal sensilla. In this study, three new putative OBP genes, MmedOBP8-10, were identified from a Microplitis mediator (Hymenoptera: Braconidae) antennal cDNA library. Quantitative real-time PCR (qRT-PCR) analysis revealed that all three of the OBP genes were expressed mainly in the antennae of adult wasps. The three OBPs were recombinantly expressed in Escherichia coli and purified by Ni ion affinity chromatography. Fluorescence competitive binding assays were performed using N-phenyl-naphthylamine as a fluorescent probe and 45 small organic compounds as competitors. These assays demonstrated that the three M. mediator OBPs can bind a broad range of odorant molecules with different binding affinities. They can bind the following ligands: nonane, farnesol, nerolidol, nonanal, β-ionone, acetic ether, and farnesene. In a Y-tube assay with these ligands as odor stimuli and paraffin oil as a control, all ligands, except nerolidol and acetic ether, were able to elicit behavioral responses in adult M. mediator. The wasps were significantly attracted to β-ionone, nonanal, and farnesene and repelled by nonane and farnesol. The results of this work provide insight into the chemosensory functions of the OBPs in M. mediator.

  5. High-throughput determination of biochemical oxygen demand (BOD) by a microplate-based biosensor.

    PubMed

    Pang, Hei-Leung; Kwok, Nga-Yan; Chan, Pak-Ho; Yeung, Chi-Hung; Lo, Waihung; Wong, Kwok-Yin

    2007-06-01

    The use of the conventional 5-day biochemical oxygen demand (BOD5) method in BOD determination is greatly hampered by its time-consuming sampling procedure and its technical difficulty in the handling of a large pool of wastewater samples. Thus, it is highly desirable to develop a fast and high-throughput biosensor for BOD measurements. This paper describes the construction of a microplate-based biosensor consisting of an organically modified silica (ORMOSIL) oxygen sensing film for high-throughput determination of BOD in wastewater. The ORMOSIL oxygen sensing film was prepared by reacting tetramethoxysilane with dimethyldimethoxysilane in the presence of the oxygen-sensitive dye tris(4,7-diphenyl-1,10-phenanthroline)ruthenium-(II) chloride. The silica composite formed a homogeneous, crack-free oxygen sensing film on polystyrene microtiter plates with high stability, and the embedded ruthenium dye interacted with the dissolved oxygen in wastewater according to the Stern-Volmer relation. The bacterium Stenotrophomonas maltophilia was loaded into the ORMOSIL/ PVA composite (deposited on the top of the oxygen sensing film) and used to metabolize the organic compounds in wastewater. This BOD biosensor was found to be able to determine the BOD values of wastewater samples within 20 min by monitoring the dissolved oxygen concentrations. Moreover, the BOD values determined by the BOD biosensor were in good agreement with those obtained by the conventional BOD5 method.

  6. 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

  7. 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.

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

    PubMed

    Clerc, Pascaline; Polster, Brian M

    2012-01-01

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

  9. 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

  10. 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.

  11. 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

  12. 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

  13. 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.

  14. A novel microplate-based screening strategy to assess the cellulolytic potential of Trichoderma strains.

    PubMed

    Cianchetta, Stefano; Galletti, Stefania; Burzi, Pier Luigi; Cerato, Claudio

    2010-10-15

    Bioconversion of lignocellulosic biomass to fuel requires a hydrolysis step to obtain fermentable sugars, generally accomplished by fungal enzymes. An assorted library of cellulolytic microbial strains should facilitate the development of optimal enzyme cocktails specific for locally available feedstocks. Only a limited number of strains can be simultaneously assayed in screening based on large volume cultivation methods, as in shake flasks. This study describes a miniaturization strategy aimed at allowing parallel assessment of large numbers of fungal strains. Trichoderma strains were cultivated stationary on microcrystalline cellulose using flat bottom 24-well plates containing an agarized medium. Supernatants obtained by a rapid centrifugation step of the whole culture plates were evaluated for extracellular total cellulase activity, measured as filter paper activity, using a microplate-based assay. The results obtained were consistent with those observed in shake-flask experiments and more than 300 Trichoderma strains were accordingly characterized for cellulase production. Five strains, displaying on shake-flasks at least 80% of the activity shown by the hyper-cellulolytic mutant Trichoderma Rut-C30, were correctly recognized by the screening on 24-well plates, demonstrating the feasibility of this approach. Cellulase activity distribution for the entire Trichoderma collection is also reported. One strain (T. harzianum Ba8/86) displayed the closest profile to the reference strain Rut-C30 in time course experiments. The method is scalable and addresses a major bottleneck in screening programs, allowing small-scale parallel cultivation and rapid supernatant extraction. It can also be easily integrated with high-throughput enzyme assays and could be suitable for automation.

  15. 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.

  16. Hydrolytic potential of Trichoderma sp. strains evaluated by microplate-based screening followed by switchgrass saccharification.

    PubMed

    Cianchetta, Stefano; Galletti, Stefania; Burzi, Pier Luigi; Cerato, Claudio

    2012-05-10

    Bioconversion of lignocellulosic biomass to fuel requires a hydrolysis step to obtain fermentable sugars, generally accomplished by fungal enzymes. Large-scale screening of different microbial strains would provide optimal enzyme cocktails for any target feedstock. The aim of this study was to screen a large collection of Trichoderma sp. strains for the hydrolytic potential towards switchgrass (Panicum virgatum L.). Strains were cultivated in a small-scale system and assayed in micro-plates for xylanase and cellulase activities. The population distributions of these traits are reported after growth on switchgrass in comparison with cellulose. The distribution profiles suggest that the growth on switchgrass strongly promotes xylanase production. The IK4 strain displayed the highest xylanase activity after growth on switchgrass (133U/mL). Enzymes (10FPU/g substrate) from IK4 were compared with those from 2 cellulolytic Trichoderma strains and a commercial enzyme in saccharification time-course experiments on untreated and pretreated switchgrass and on an artificial substrate. Samples were analysed by DNS assay and by an oxygraphic method for sugar equivalent or glucose concentration. On the untreated substrate, IK4 enzymes even outperformed a 5-fold load of commercial enzyme, suggesting that xylanase or accessory enzymes are a limiting factor on this type of recalcitrant substrate. On the other substrates, IK4 preparations showed intermediate behaviour if compared with the commercial enzyme at 10FPU/g substrate and at 5-fold load. IK4 also nearly halved the time to release 50% of the hydrolysable sugar equivalents (T(50%)), with respect to the other preparations at the same enzymatic load. DNS assay and oxygraphic method gave highly correlated results for the 3 saccharified substrates. The study suggests that accessory enzymes like xylanase play a key role in improving the performance of cellulase preparations on herbaceous lignocellulosic feedstocks like switchgrass.

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

    NASA Astrophysics Data System (ADS)

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

    2016-10-01

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

  18. GPS-Constrained Microplate Kinematics and Plio-Pleistocene Tectonic Evolution of the North Anatolian Fault and North Aegean Sea

    NASA Astrophysics Data System (ADS)

    Thatcher, W.

    2004-12-01

    Emerging evidence from Global Position System (GPS) survey measurements in the Aegean and elsewhere suggests that present-day active continental deformation occurs largely due to the relative motions of a small number of rigid blocks or microplates. However, it is not universally agreed whether the continental microplate description of the GPS data is superior to other proposed models, nor is it clear whether present-day movement patterns can be usefully extrapolated into the geologic past. Here I examine the known deformation history of the North Aegean over the past ~10 Ma and compare it with predictions based on the present-day microplate model. Agreement provides independent support for the GPS-based model and demonstrates its value in bettering our understanding of Aegean tectonics. If we knew nothing about late Cenozoic North Aegean tectonics and provisionally assumed the correctness of the Aegean microplate model of Nyst & Thatcher [2004 JGR], we would predict several features of the tectonic evolution that accord with geologic evidence. First, the North Aegean Sea would be created by extension due to SSW motion of the South Aegean and concomitant CW rotation of central Greece during the past 10 Ma. The same kinematic process would cause extension to be succeeded by strike-slip motion as the `ridge-transform-ridge' triple junction migrates WSW 24 km/Ma and the North Anatolian fault propagates into the region. The Plio-Pleistocene history of the North Aegean shows these same general features. Drilling and seismic imaging document the existence of young (< 10 Ma) and thick (up to 6 km) sedimentary sequences attributed to crustal extension by a factor of 3-4. Seismic profiling and bathymetric mapping show a mesh of roughly orthogonal faults with dip-slip offsets. Structural studies of sub-aerial exposures of these faults suggest an earlier episode of extension was followed by predominantly strike-slip motions. The plate kinematic reconstruction of late Cenozoic

  19. 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

  20. 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

  1. 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.

  2. 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.

  3. 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

  4. 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

  5. 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

  6. 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

  7. U-Pb isotopic evidence for accretion of a continental microplate in the Zalm region of the Saudi Arabian Shield

    USGS Publications Warehouse

    Stacey, J.S.; Agar, R.A.

    1986-01-01

    U-Pb zircon age determinations show that this "Andean" continental margin developed before about 720 Ma, and emplacement of calc-alkaline plutonic rocks continued until about 690 Ma. During the period 690-640 Ma, the continental Afif microplate collided with the Asir terrane as part of the Nabitah orogeny. At approximately 640 Ma ago, the Najd strike-slip orogen commenced with a dextral phase that controlled the emplacement of granitic plutons as well as the development of a series of large pull-apart grabens. Some of these grabens were floored by new oceanic crust and were filled with volcano-sedimentary rocks of the Bani Ghayy group. Subsequently, the Najd fault system changed to sinistral strike slip motion at about 620 Ma ago.

  8. 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

  9. 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.

  10. Biofilm formation by environmental isolates of Salmonella and their sensitivity to natural antimicrobials

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We evaluated 15 Salmonella isolates; S. Derby (2), S. Infantis (4), and S. Typhimurium (9) from conventional swine farm environment (soil and lagoon) for biofilm formation. Biofilm forming ability was determined by 96-well microtitre plate Crystal-Violet and Minimum Biofilm Eradication Concentration...

  11. 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

  12. 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

  13. 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

  14. 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

  15. 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.

  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. Sensitive fluorescent microplate bioassay using recombinant Escherichia coli with multiple promoter-reporter units in tandem for detection of arsenic.

    PubMed

    Tani, Chiaki; Inoue, Koichi; Tani, Yuri; Harun-ur-Rashid, Md; Azuma, Norihiro; Ueda, Shunsaku; Yoshida, Kazuyuki; Maeda, Isamu

    2009-11-01

    Genetically modified bacterial biosensors can detect specific environmental compounds. Here, we attempted to establish a fluorescent microplate method to detect arsenic using recombinant Escherichia coli cells transformed with plasmids harboring three tandem copies of the ars promoter/operator-the gene for green fluorescent protein (gfp). In the biosensors, one copy of arsR, whose transcription is autoregulated by the ars promoter/operator and ArsR in the genome of E. coli, was placed in trans in another plasmid under the control of isopropyl-1-thio-beta-D-galactopyranoside-inducible promoter. First, this manipulation enabled regulation of the arsR expression at an adequate level. Second, the copy number of reporter unit also affected signal and noise. When the plasmid harboring three copies of the reporter unit was used, the signal-to-noise ratio doubled and the detection limit decreased from 20 to 7.5 microg L(-1) As(III), compared to the use of the plasmid harboring one copy of the ars promoter/operator-arsR-gfp. Thus, segregation of arsR from the ars promoter/operator-gfp using two plasmids is effective in regulating the signal-to-noise ratio and the detection limit with the different functions.

  18. Late Pleistocene and Holocene uplift history of Cyprus: implications for active tectonics along the southern margin of the Anatolian microplate

    USGS Publications Warehouse

    Harrison, R.W.; Tsiolakis, E.; Stone, B.D.; Lord, A.; McGeehin, J.P.; Mahan, S.A.; Chirico, P.

    2013-01-01

    The nature of the southern margin of the Anatolian microplate during the Neogene is complex, controversial and fundamental in understanding active plate-margin tectonics and natural hazards in the Eastern Mediterranean region. Our investigation provides new insights into the Late Pleistocene uplift history of Cyprus and the Troodos Ophiolite. We provide isotopic (14C) and radiogenic (luminescence) dates of outcropping marine sediments in eastern Cyprus that identify periods of deposition during marine isotope stages (MIS) 3, 4, 5 and 6. Past sea-levels indicated by these deposits are c. 95±25 m higher in elevation than estimates of worldwide eustatic sea-level. An uplift rate of c. 1.8 mm/year and possibly as much as c. 4.1 mm/year in the past c. 26–40 ka is indicated. Holocene marine deposits also occur at elevations higher than those expected for past SL and suggest uplift rates of c. 1.2–2.1 mm/year. MIS-3 marine deposits that crop out in southern and western Cyprus indicate uniform island-wide uplift. We propose a model of tectonic wedging at a plate-bounding restraining bend as a mechanism for Late Pleistocene to Holocene uplift of Cyprus; uplift is accommodated by deformation and seismicity along the margins of the Troodos Ophiolite and re-activation of its low-angle, basal shear zone.

  19. 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

  20. 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

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

    PubMed

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

    2016-01-01

    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.

  3. 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

  4. A Microplate-Based Nonradioactive Protein Synthesis Assay: Application to TRAIL Sensitization by Protein Synthesis Inhibitors

    PubMed Central

    Henrich, Curtis J.

    2016-01-01

    Non-radioactive assays based on incorporation of puromycin into newly synthesized proteins and subsequent detection using anti-puromycin antibodies have been previously reported and well-validated. To develop a moderate- to high-throughput assay, an adaptation is here described wherein cells are puromycin-labeled followed by simultaneously probing puromycin-labeled proteins and a reference protein in situ. Detection using a pair of near IR-labeled secondary antibodies (InCell western, ICW format) allows quantitative analysis of protein synthesis in 384-well plates. After optimization, ICW results were compared to western blot analysis using cycloheximide as a model protein synthesis inhibitor and showed comparable results. The method was then applied to several protein synthesis inhibitors and revealed good correlation between potency as protein synthesis inhibitors to their ability to sensitize TRAIL-resistant renal carcinoma cells to TRAIL-induced apoptosis. PMID:27768779

  5. 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.

  6. Validation of the Immunalysis microplate ELISA for the detection of buprenorphine and its metabolite norbuprenorphine in urine.

    PubMed

    Miller, Eleanor I; Torrance, Hazel J; Oliver, John S

    2006-03-01

    The purpose of this study was to validate the Immunalysis Buprenorphine Microplate enzyme-linked immunosorbent assay (ELISA) for the detection of buprenorphine in urine samples. Sixty-nine urine samples were obtained from volunteers on the Subutex treatment program and from routine samples submitted to the laboratory for buprenorphine testing. For ELISA analysis, samples were diluted 1:10 with K(2)HPO(4) (0.1M, pH 7.0). The limit of detection was calculated as 0.5 ng/mL buprenorphine. The intra-assay and interday precision was 3.8% (n = 10) and 8.6% (n = 50) respectively at 1 ng/mL buprenorphine. At a low concentration of norbuprenorphine (1 ng/mL), the immunoassay demonstrated a cross-reactivity of 78%. A higher cross-reactivity of 116% was observed at a higher concentration of norbuprenorphine (10 ng/mL). Dihydrocodeine, codeine, tramadol, morphine, propoxyphene, methadone, and EDDP were tested at concentrations of 10 ng/mL and 10,000 ng/mL and demonstrated no cross-reactivity with the assay. For liquid chromatography-tandem mass spectrometry (LC-MS-MS), deuterated internal standard mixture, 1M acetate buffer (pH 5.0), and b-glucuronidase were added to the standards and samples, which were then incubated for 3 h at 60 degrees C. After incubation, 3 mL K(2)HPO(4) (0.1M, pH 6.0) was added and the pH altered to pH 6.0 using 1M KOH. Buprenorphine and norbuprenorphine were subsequently extracted by solid-phase. Twenty-one samples were confirmed positive and 48 samples were confirmed negative by LC-MS-MS. Using a cut-off value of 0.5 ng/mL buprenorphine, the immunoassay demonstrated a sensitivity and specificity of 100%. PMID:16620543

  7. 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

  8. A transcriptome analysis suggests apoptosis-related signaling pathways in hemocytes of Spodoptera litura after parasitization by Microplitis bicoloratus.

    PubMed

    Li, Ming; Pang, Zunyu; Xiao, Wei; Liu, Xinyi; 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

  9. [Automatic reading of ABO and Rh groups on microplates using FMC medium and an IBG Systems reader].

    PubMed

    Mauri, J; Maymo, R M; Pérez, M A; Yusta, V; Mas, J; Puig, L

    1991-06-01

    We present in this paper our experience in the routine use of an automatic reader for microtiter plates (IBG Systems). A total of 2044 samples from blood donors have been tested for ABO (haematic and seric) and Rh (D antigen) blood group typing. The red blood cell samples have been tested against monoclonal anti-A, anti-B, anti-AB and anti-Rh 1 (D) sera (using two different anti-D reagents). As a negative control, 3% albumin solution was employed. In order to determine the seric groups, the plasma samples were tested against A1, A2, B and 0 cells. In all cases the red cells were suspended in Ficoll 400R-Methyl cellulose (FMC), and 0.01 Bromelin was added to red blood cell samples to be typed. The obstacles in the automatic readings were mainly solved by visual reading of the microplates. In the 2,044 samples analysed with the automatic reader, 19 discrepancies (0.94%) were found in ABO typing. In all cases the error was in the seric blood group. Three false positive reactions were found with 0 red cells. Two false negative with B red cells, interpreted by the automatic reader as doubtful results. Three positive readings were detected, in one case the positivity was due to alloantibodies (C + D specificities), the other two cases were autoagglutinations. False negative reactions were found, in 11 cases, 8 of them due to lipaemia and the remaining three were haemolysed plasma samples. It should be stressed that out of 19 cases of discrepancy, only 5 (0.24%) were due to the automatic reader while the remaining were due, to sample troubles.(ABSTRACT TRUNCATED AT 250 WORDS)

  10. The Dinaric Faults System: Large-scale Structure And Rates Of Slip Of The Transpressive Northeastern Boundary Of The Adria Microplate

    NASA Astrophysics Data System (ADS)

    Moulin, A.

    2015-12-01

    Located at the northeastern corner of the Adria microplate, the Alps-Dinarides junction represents a key region for understanding how the microplate interacts with stable Europe. While the active tectonics of the alpine part of this area is relatively well-constrained, little is known about the present-day deformation imposed by the Adria rotation across the Dinarides. Through morpho-tectonic analysis based on satellite and aerial images, accurate topographical maps and digital elevation models (including LiDAR) combined with field investigations, we have mapped in details the active faults of the Northern Dinarides and the eastern part of the Southern Alps. Based on this mapping and on geological data, we have identified forty cumulative tectonic displacements ranging from few meters to several kilometres allowing to decipher how deformations have affected this area over the Pliocene to the present-day situation. The Late Pleistocene period was mainly investigated using 36Cl-exposure dating of displaced geomorphic markers combined with the existing chronologies of the glacial-interglacial landscape evolution in the area (e.g Monegato et al., 2007 ; Fontana et al., 2008). The presented results yield 3.1±0.9 mm/yr of active right-lateral faulting along the Northern Dinarides. The Adria rotation models (e.g Calais et al., 2002 ; D'Agostino et al., 2008; Weber et al., 2010) and the present knowledge of the lithospheric structure of the area (Brückl et al., 2010 ; Sumanovac et al., 2009) along with our results corroborates that the Northern Dinarides probably represents the main eastern boundary of the Adria microplate.

  11. 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.

  12. Development, Optimization, and Validation of a Microplate Bioassay for Relative Potency Determination of Linezolid Using a Design Space Concept, and its Measurement Uncertainty.

    PubMed

    Saviano, Alessandro Morais; Francisco, Fabiane Lacerda; Ostronoff, Celina Silva; Lourenço, Felipe Rebello

    2015-01-01

    The aim of this study was to develop, optimize, and validate a microplate bioassay for relative potency determination of linezolid in pharmaceutical samples using quality-by-design and design space approaches. In addition, a procedure is described for estimating relative potency uncertainty based on microbiological response variability. The influence of culture media composition was studied using a factorial design and a central composite design was adopted to study the influence of inoculum proportion and triphenyltetrazolium chloride in microbial growth. The microplate bioassay was optimized regarding the responses of low, medium, and high doses of linezolid, negative and positive controls, and the slope, intercept, and correlation coefficient of dose-response curves. According to optimization results, design space ranges were established using: (a) low (1.0 μg/mL), medium (2.0 μg/mL), and high (4.0 μg/mL) doses of pharmaceutical samples and linezolid chemical reference substance; (b) Staphylococcus aureus ATCC 653 in an inoculum proportion of 10%; (c) antibiotic No. 3 culture medium pH 7.0±0.1; (d) 6 h incubation at 37.0±0.1ºC; and (e) addition of 50 μL of 0.5% (w/v) triphenyltetrazolium chloride solution. The microplate bioassay was linear (r2=0.992), specific, precise (repeatability RSD=2.3% and intermediate precision RSD=4.3%), accurate (mean recovery=101.4%), and robust. The overall measurement uncertainty was reasonable considering the increased variability inherent in microbiological response. Final uncertainty was comparable with those obtained with other microbiological assays, as well as chemical methods. PMID:26525245

  13. 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

  14. Development, Optimization, and Validation of a Microplate Bioassay for Relative Potency Determination of Linezolid Using a Design Space Concept, and its Measurement Uncertainty.

    PubMed

    Saviano, Alessandro Morais; Francisco, Fabiane Lacerda; Ostronoff, Celina Silva; Lourenço, Felipe Rebello

    2015-01-01

    The aim of this study was to develop, optimize, and validate a microplate bioassay for relative potency determination of linezolid in pharmaceutical samples using quality-by-design and design space approaches. In addition, a procedure is described for estimating relative potency uncertainty based on microbiological response variability. The influence of culture media composition was studied using a factorial design and a central composite design was adopted to study the influence of inoculum proportion and triphenyltetrazolium chloride in microbial growth. The microplate bioassay was optimized regarding the responses of low, medium, and high doses of linezolid, negative and positive controls, and the slope, intercept, and correlation coefficient of dose-response curves. According to optimization results, design space ranges were established using: (a) low (1.0 μg/mL), medium (2.0 μg/mL), and high (4.0 μg/mL) doses of pharmaceutical samples and linezolid chemical reference substance; (b) Staphylococcus aureus ATCC 653 in an inoculum proportion of 10%; (c) antibiotic No. 3 culture medium pH 7.0±0.1; (d) 6 h incubation at 37.0±0.1ºC; and (e) addition of 50 μL of 0.5% (w/v) triphenyltetrazolium chloride solution. The microplate bioassay was linear (r2=0.992), specific, precise (repeatability RSD=2.3% and intermediate precision RSD=4.3%), accurate (mean recovery=101.4%), and robust. The overall measurement uncertainty was reasonable considering the increased variability inherent in microbiological response. Final uncertainty was comparable with those obtained with other microbiological assays, as well as chemical methods.

  15. Possible microplate generation at RRR triple junctions due to the non-circular finite motion of plates relative to each other

    NASA Astrophysics Data System (ADS)

    Cronin, V. S.

    2012-12-01

    First generation ideas of the kinematic stability of triple junctions lead to the common belief that the geometry of ridge-ridge-ridge (RRR) triple junctions remains constant over time under conditions of symmetric spreading. Given constant relative motion between each plate pair -- that is, the pole of plate relative motion is fixed to both plates in each pair during finite motion, as assumed in many accounts of plate kinematics -- there would be no boundary mismatch at the triple junction and no apparent kinematic reason why a microplate might develop there. But if, in a given RRR triple junction, the finite motion of one plate as observed from the other plate is not circular (as is generally the case, given the three-plate problem of plate kinematics), the geometry of the ridges and the triple junction will vary with time (Cronin, 1992, Tectonophys 207, 287-301). To explore the possible consequences of non-circular finite motion between plates at an RRR triple junction, a simple model was coded based on the cycloid finite-motion model (e.g., Cronin, 1987, Geology 15, 1006-1009) using NNR-MORVEL56 velocities for individual plates (Argus et al., 2011, G3 12, doi: 10.1029/2011GC003751). Initial assumptions include a spherical Earth, symmetric spreading, and constant angular velocities during the modeled finite time interval. The assumed-constant angular velocity vectors constitute a reference frame for observing finite plate motion. Typical results are [1] that the triple junction migrates relative to a coordinate system fixed to the angular-velocity vectors, [2] ridge axes rotates relative to each other, and [3] a boundary mismatch develops at the synthetic triple junction that might result in microplate nucleation. In a model simulating the Galapagos triple junction between the Cocos, Nazca and Pacific plates whose initial state did not include the Galapagos microplate, the mismatch gap was as much as ~3.4 km during 3 Myr of model displacement (see figure). The

  16. Epstein-Barr virus early antigen titer by immunofluorescence in microplates. A new semi-automated method based on microtiter system.

    PubMed

    Favart, A M; Lamy, M E; Allemeersch, D; Burtonboy, G; Vanoverschelde, J

    1978-11-17

    For the titration of antibodies directed against Epstein-Barr Virus early antigen (EBV-EA) we describe a method in microplates with the Microtiter System (Cooke Engineering Co., Alexandria, Va). Using this technique, which allows rapid epidemiological investigations, we have titered anti-EBV-EA antibodies in different groups of patients and controls, and particularly in group which had recent contact with infectious mononucleosis. In this group the percentage of individuals having antibodies directed against the EBV-EA antigen was significantly higher than in the group of controls.

  17. 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

  18. A new barrier to dispersal trapped old genetic clines that escaped the Easter Microplate tension zone of the Pacific vent mussels.

    PubMed

    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.

  19. 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 Reaction (RX) Plate method, a modification of the RX tube designed by Gailey et al. (2004), consolidates pre-enrichment (buffered peptone water or GN Hajna), enrichm...

  20. 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.

  1. 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...

  2. 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...

  3. 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.

  4. Inter-seismic deformation and plate coupling along the Andaman micro-plate margin: geodetic constraints using 1996-2004 GPS data.

    NASA Astrophysics Data System (ADS)

    Earnest, A.; Puchakayala, J.; Rajendran, C. C. P.; Rajendran, K.

    2014-12-01

    Oblique convergence of the Andaman microplate with the Sunda margin results in permanent deformation within the overriding plate and had generated giant plate-boundary ruptures like the 2004 Mw 9.3 Sumatra- Andaman earthquake. Inter-seismic upper plate deformation of this part of the subduction zone remains poorly constrained due to lack of availability of spatially well distributed data. Through this study we are reanalyzing the available GPS geodetic data sets collected by various agencies over different times between 1996-2004 to constrain the pre-earthquake convergence values using a consistent reference frame to determine the crustal deformation in the Andaman Nicobar region, to infer strain rates in the overriding plate, how Andaman microplate was moving relative to Indian plate and what was the extent of plate locking. We will also discuss the details on spatial and temporal variations of convergence rates and variations in plate coupling. To quantify the change in coupling, we calculated the interseismic surface deformation using Okada's formulation, in which locked faults are modelled as dislocations in a halfspace. We defined the subduction zone geometry as three elastically deforming blocks (India, Andaman fore-arc, and Sunda) separated by two faults: the West Andaman Fault and the Sumatra-Andaman megathrust. The bennioff-zone dip and orientation is re-defined using the recently relocated epicentral solutions with higher accuracy and from the slip-distribution models constrained from the various co-seismic geodetic offsets reported from the near-field and far-field GPS sites . The rigid-body motion of each block is specified by a pole of rotation. We modelled the variation in coupling by specifying for each node and integrating the slip deficit over the fault surfaces. The West Andaman Fault is modeled as a vertical strike- lip fault, locked to a depth of ~20 km, with a dextral slip.

  5. 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

  6. 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

  7. A Small Ocean Rift Leads to a New View of the Galapagos Microplate: the Incipient Rift at 2 Degrees North, East of the East Pacific Rise

    NASA Astrophysics Data System (ADS)

    Klein, E. M.; Smith, D. K.; Williams, C. M.; Schouten, H.

    2004-12-01

    The Galapagos microplate (GMP) shares a complex plate boundary configuration with the surrounding Cocos, Nazca and Pacific plates. While the configuration of the microplate's southern boundary with the Nazca plate is relatively well understood, the nature of its northern boundary with the Cocos plate has remained more elusive. Work by Lonsdale and co-workers (1988; 1992) identified an "incipient" spreading center east of and orthogonal to the East Pacific Rise (EPR) at 2deg40minN, forming a portion of the northern boundary of the GMP. They described this spreading center as a slowly diverging, westward propagating rift, verging toward the EPR. In 2002, we mapped and sampled a broad region centered on the IR, extending ~140 km east of the EPR. Various geological, geophysical and rock sampling tools were used, including Seabeam2000 bathymetry and side-scan (amplitude); towed magnetometer; bottom photography (14 camera tows using the WHOI camera system); water column hydrothermal surveying and rock sampling. Our recent analysis of the bathymetric, side-scan and magnetic data suggests that the IR forms a lozenge-shaped feature, which tapers both westward toward the EPR and eastward, away from the EPR. Within the eastern half of the lozenge, there is a sinuous trough that trends ~100° to the southeast, along which reflection amplitudes indicate high-reflectivity consistent with relatively sparse sediment cover. The eastern end of this trough clearly cuts north-south-oriented abyssal hills and then dies out in the vicinity of 101deg29minW. The photographic results show sparsely sedimented lavas, often with delicate ornamentation and basaltic glass, with local in-filling of sediment between pillows. In a number of photographs, particularly along the eastern portion of the IR, lavas appear to emanate from local fissures with southeast-northwest orientations. Taken together, these data suggest that the IR is magmatically active along its length and has propagated eastward

  8. 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

  9. 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.

  10. 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.

  11. 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.

  12. 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

  13. Titration of varicella-zoster virus DNA in throat swabs from varicella patients by combined use of PCR and microplate hybridization.

    PubMed

    Hondo, R; Ito, S; Inouye, S

    1995-01-01

    We devised a simple procedure for titration of varicella-zoster virus (VZV) DNA in throat swabs from varicella patients. DNA which was extracted from throat swabs, together with known copy numbers of a cloned VZV DNA fragment, were 10-fold serially diluted and used as template in PCR. The PCR products, after heat denaturation, again serially diluted in 1.5 M NaCl and adsorbed to microplate wells. Then, biotin-labeled DNA probes were hybridized with the immobilized DNA. The hybridization signal was produced by streptavidin-conjugated beta-galactosidase and a fluorogenic enzyme substrate. By comparing the titration curves of a clinical specimen with those of the cloned fragment, of which detection limit was about 10 copies, we estimated the copy numbers of VZV DNA in the specimen. With this technique, we evaluated the degree of potential contagiousness of the patient along the course of infection: we found that varicella patients possessed highest quantity of VZV DNA in the throat on the first day of illness.

  14. 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

  15. 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.

  16. High-throughput method for determination of apolipoprotein E genotypes with use of restriction digestion analysis by microplate array diagonal gel electrophoresis.

    PubMed

    Bolla, M K; Haddad, L; Humphries, S E; Winder, A F; Day, I N

    1995-11-01

    Molecular epidemiological research has identified the association of a common apolipoprotein E (apo E) isoform (E4 as opposed to E3), with risk both of coronary artery disease and of Alzheimer dementia. In addition, the role of apo E genotype (usually E2/E2) in Type III hyperlipidemia is well known. However, both for diagnostic and research purposes, apo E genotyping is cumbersome. The preferred approach is electrophoretic sizing of restriction digestion fragments, enabling simultaneous analysis of the two codons (112 and 158) that represent the six common genotypes (E2/E2; E2/E3; E2/E4; E3/E3; E3/E4; E4/E4). However, the consequent demands of high-yield PCR, high-resolution, high-throughput electrophoresis, and sufficient detection sensitivity have left shortfalls in published protocols. In conjunction with a high-throughput electrophoresis system we described recently, microplate array diagonal gel electrophoresis (MADGE), we have constructed extensively optimized, simplified protocols for DNA isolation from mouthwash samples for PCR setup and high-yield PCR, for restriction digestion, and for subsequent MADGE gel image analysis. The integral system enables one worker to readily undertake apo E genotyping of as many as hundreds of DNA samples per day, without special equipment.

  17. Application of a microplate-based ORAC-pyrogallol red assay for the estimation ofantioxidant capacity: First Action 2012.03.

    PubMed

    Ortiz, Rocío; Antilén, Mónica; Speisky, Hernán; Aliaga, Margarita E; López-Alarcón, Camilo; Baugh, Steve

    2012-01-01

    A method was developed for microplate-based oxygen radicals absorbance capacity (ORAC) using pyrogallol red (PGR) as probe (ORAC-PGR). The method was evaluated for linearity, precision, and accuracy. In addition, the antioxidant capacity of commercial beverages, such as wines, fruit juices, and iced teas, was measured. Linearity of the area under the curve (AUC) versus Trolox concentration plots was [AUC = (845 +/- 110) + (23 +/- 2) [Trolox, microM]; R = 0.9961, n = 19]. Analyses showed better precision and accuracy at the highest Trolox concentration (40 microM) with RSD and recovery (REC) values of 1.7 and 101.0%, respectively. The method also showed good linearity for red wine [AUC = (787 +/- 77) + (690 +/- 60) [red wine, microL/mL]; R = 0.9926, n = 17], precision and accuracy with RSD values from 1.4 to 8.3%, and REC values that ranged from 89.7 to 103.8%. Red wines showed higher ORAC-PGR values than white wines, while the ORAC-PGR index of fruit juices and iced teas presented a wide range of results, from 0.6 to 21.6 mM of Trolox equivalents. Product-to-product variability was also observed for juices of the same fruit, showing the differences between brands on the ORAC-PGR index.

  18. RNA interference silences Microplitis demolitor bracovirus genes and implicates glc1.8 in disruption of adhesion in infected host cells.

    PubMed

    Beck, Markus; Strand, Michael R

    2003-09-30

    The family Polydnaviridae consists of ds-DNA viruses that are symbiotically associated with certain parasitoid wasps. PDVs are transmitted vertically but also are injected by wasps into hosts where they cause several physiological alterations including immunosuppression. The PDV genes responsible for mediating immunosuppression and other host alterations remain poorly characterized in large measure because viral mutants cannot be produced to study gene function. Here we report the use of RNA interference (RNAi) to specifically silence the glc1.8 and egf1.0 genes from Microplitis demolitor bracovirus (MdBV) in High Five cells derived from the lepidopteran Trichoplusia ni. Dose-response studies indicated that MdBV infects High Five cells and blocks the ability of these cells to adhere to culture plates. This response was very similar to what occurs in two classes of hemocytes, granular cells, and plasmatocytes, after infection by MdBV. Screening of monoclonal antibody (mAb) markers that distinguish different classes of lepidopteran hemocytes indicated that High Five cells cross-react with three mAbs that recognize granular cells from T. ni. Double-stranded RNA (dsRNA) complementary to glc1.8 specifically silenced glc1.8 expression and rescued the adhesive phenotype of High Five cells. Reciprocally, dsRNA complementary to egf1.0 silenced egf1.0 expression but had no effect on adhesion. The simplicity and potency of RNAi could be extremely useful for analysis of other PDV genes.

  19. Combination of 768-well microplate array diagonal gel electrophoresis with duplex PCR of X and Y chromosome markers for quality control of epidemiological DNA banks.

    PubMed

    Huang, Shuwen; Chen, Xiao-he; Day, Ian N M

    2006-08-01

    Large DNA banks for human epidemiological studies have become an increasingly important research tool. The power of genotype-phenotype studies is dependent both on the quality of phenotyping and of genotyping and of correct linking of phenotypes to genotypes. Samples must be tracked through numerous steps between subject or patient and post-genotypic data. Only one phenotype, sex, has a perfect and binary correlation with genotype. In mixed sex studies, it may be advantageous for purposes of quality control to keep sexes mixed during the steps from acquisition to DNA bank, in order to be able to check later for sample swaps. We have designed a duplex PCR combining an amplicon from MAOA marking the X chromosome and an amplicon from DDX3Y marking the Y chromosome. We combined this with a simple economical palmtop sized 768-well microplate compatible electrophoresis system developed in-house for examination of duplex PCR products. We applied this quality control test in the validation of two DNA banks.

  20. 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

  1. 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.

  2. A polydnaviral genome of Microplitis bicoloratus bracovirus and molecular interactions between the host and virus involved in NF-κB signaling.

    PubMed

    Yu, Dong-Shuai; Chen, Ya-Bin; Li, Ming; Yang, Ming-Jun; Yang, Yang; Hu, Jian-Sheng; Luo, Kai-Jun

    2016-11-01

    Polydnaviruses (PDVs) play a critical role in altering host gene expression to induce immunosuppression. However, it remains largely unclear how PDV genes affect host genes. Here, the complete genome sequence of Microplitis bicoloratus bracovirus (MbBV), which is known to be an apoptosis inducer, was determined. The MbBV genome consisted of 17 putative double-stranded DNA circles and 179 fragments with a total size of 336,336 bp and contained 116 open reading frames (ORFs). Based on conserved domains, nine gene families were identified, of which the IκB-like viral ankyrin (vank) family included 28 members and was one of the largest families. Among the 116 ORFs, 13 MbBV genes were expressed in hemocytes undergoing MbBV-induced apoptosis and further analyzed. Three vank genes (vank86, vank92, vank101) were expressed in hemocytes collected from Spodoptera litura larvae parasitized by M. bicoloratus, in which host NF-κB/IκBs, including relish, dorsal, and cactus, were also persistently expressed. When Spli221 cells were infected with MbBV viral particles, mRNA levels of host and viral NF-κB/IκB genes were persistent and also varied in Spli221 cells undergoing virus-induced pre-apoptosis cell from 1 to 5 hours postinfection. Both were then expressed in a time-dependent expression in virus-induced apoptotic cells. These data show that viral IκB-like transcription does not inhibit host NF-κB/IκB expression, suggesting that transcription of these genes might be regulated by different mechanisms. PMID:27522588

  3. 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

  4. Ellman's-reagent-mediated regeneration of trypanothione in situ: substrate-economical microplate and time-dependent inhibition assays for trypanothione reductase.

    PubMed

    Hamilton, Chris J; Saravanamuthu, Ahilan; Eggleston, Ian M; Fairlamb, Alan H

    2003-02-01

    Trypanothione reductase (TryR) is a key enzyme involved in the oxidative stress management of the Trypanosoma and Leishmania parasites, which helps to maintain an intracellular reducing environment by reduction of the small-molecular-mass disulphide trypanothione (T[S](2)) to its di-thiol derivative dihydrotrypanothione (T[SH](2)). TryR inhibition studies are currently impaired by the prohibitive costs of the native enzyme substrate T[S](2). Such costs are particularly notable in time-dependent and high-throughput inhibition assays. In the present study we report a protocol that greatly decreases the substrate quantities needed for such assays. This is achieved by coupling the assay with the chemical oxidant 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), which can rapidly re-oxidize the T[SH](2) product back into the disulphide substrate T[S](2), thereby maintaining constant substrate concentrations and avoiding deviations from rate linearity due to substrate depletion. This has enabled the development of a continuous microplate assay for both classical and time-dependent TryR inhibition in which linear reaction rates can be maintained for 60 min or more using minimal substrate concentrations (<1 microM, compared with a substrate K (m) value of 30 microM) that would normally be completely consumed within seconds. In this manner, substrate requirements are decreased by orders of magnitude. The characterization of a novel time-dependent inhibitor, cis -3-oxo-8,9b-bis-(N(1)-acrylamidospermidyl)-1,2,3,4,4a,9b-hexahydrobenzofuran (PK43), is also described using these procedures.

  5. 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

  6. 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.

  7. Formation of biofilm by Listeria monocytogenes ATCC 19112 at different incubation temperatures and concentrations of sodium chloride.

    PubMed

    Lee, H Y; Chai, L C; Pui, C F; Mustafa, S; Cheah, Y K; Nishibuchi, M; Radu, S

    2013-01-01

    Biofilm formation can lead to various consequences in the food processing line such as contamination and equipment breakdowns. Since formation of biofilm can occur in various conditions; this study was carried out using L. monocytogenes ATCC 19112 and its biofilm formation ability tested under various concentrations of sodium chloride and temperatures. Cultures of L. monocytogenes ATCC 19112 were placed in 96-well microtitre plate containing concentration of sodium chloride from 1-10% (w/v) and incubated at different temperature of 4 °C, 30 °C and 45 °C for up to 60 h. Absorbance reading of crystal violet staining showed the density of biofilm formed in the 96-well microtitre plates was significantly higher when incubated in 4 °C. The formation of biofilm also occurs at a faster rate at 4 °C and higher optical density (OD 570 nm) was observed at 45 °C. This shows that storage under formation of biofilm that may lead to a higher contamination along the processing line in the food industry. Formation of biofilm was found to be more dependent on temperature compared to sodium chloride stress.

  8. 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

  9. Biofilm formation and acyl homoserine lactone production in Hafnia alvei isolated from raw milk.

    PubMed

    Viana, Eliseth Souza; Campos, Maria Emilene Martino; Ponce, Adriana Reis; Mantovani, Hilário Cuquetto; Vanetti, Maria Cristina Dantas

    2009-01-01

    The objective of this study was to detect the presence of acyl homoserine lactones (AHLs), signal molecules of the quorum sensing system in biofilm formed by Hafnia alvei strains. It also evaluated the effect of synthetic quorum sensing inhibitors in biofilm formation. AHLs were assayed using well diffusion techniques, thin layer chromatography (TLC) and detection directly in biofilm with biomonitors. The extracts obtained from planktonic and sessile cell of H. alvei induced at least two of three monitor strains evaluated. The presence of AHLs with up to six carbon atoms was confirmed by TLC. Biofilm formation by H. alvei was inhibited by furanone, as demonstrated by 96-well assay of crystal violet in microtitre plates and by scanning electron microscopy. The H. alvei 071 hall mutant was deficient in biofilm formation. All these results showed that the quorum sensing system is probably involved in the regulation of biofilm formation by H. alvei. PMID:20140298

  10. Effect of tunicamycin on Candida albicans biofilm formation and maintenance

    PubMed Central

    Pierce, Christopher G.; Thomas, Derek P.; López-Ribot, José L.

    2009-01-01

    Background Candida albicans is a common opportunistic pathogen of the human body and is the frequent causative agent of candidiasis. Typically, these infections are associated with the formation of biofilms on both host tissues and implanted biomaterials. As a result of the intrinsic resistance of C. albicans biofilms to most antifungal agents, new strategies are needed to combat these infections. Methods Here we have used a 96-well microtitre plate model of C. albicans biofilm formation to study the inhibitory effect of tunicamycin, a nucleoside antibiotic that inhibits N-linked glycosylation affecting cell wall and secreted proteins, on C. albicans biofilm formation. A proteomic approach was used to study the effect of tunicamycin on levels of glycosylation of key secreted mannoproteins in the biofilm matrix. Results Our results revealed that physiological concentrations of tunicamycin displayed significant inhibitory effects on biofilm development and maintenance, while not affecting overall cell growth or morphology. However, tunicamycin exerted a minimal effect on fully mature, pre-formed C. albicans biofilms. Conclusions The effect of tunicamycin on the C. albicans biofilm mode of growth demonstrates the importance of N-linked glycosylation in the developmental stages of biofilm formation. In addition, our results indicate that N-linked glycosylation represents an attractive target for the development of alternative strategies for the prevention of biofilm formation by this important pathogenic fungus. PMID:19098294

  11. Development of a technology for automation and miniaturization of protein crystallization.

    PubMed

    Mueller, U; Nyarsik, L; Horn, M; Rauth, H; Przewieslik, T; Saenger, W; Lehrach, H; Eickhoff, H

    2001-01-23

    The usage of standard 96 well microplates for the screening of crystallization conditions of recombinant proteins offers several advantages when compared to commonly used crystallization plate formats. The adoption of robotic technology for plate and glass slide preparation within a "hanging drop" vapour diffusion crystallization experiment enables to work with an increased throughput at reduced costs. In addition to commercial pipetting devices with a 96-channel aspirator/dispenser, solenoid ink-jet technology was applied to form 250 nl droplets with a diameter of approximately 1 mm. This allows miniaturization of crystallization screening set-ups with an estimated ten-fold cost reduction when compared to commonly used 24 well plates.

  12. 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.

  13. Kinetics of advanced glycation end products formation on bovine serum albumin with various reducing sugars and dicarbonyl compounds in equimolar ratios.

    PubMed

    Luers, Lars; Rysiewski, Karolina; Dumpitak, Christian; Birkmann, Eva

    2012-04-01

    Reducing sugars and reactive dicarbonyl compounds play a major role in glycation of proteins in vivo. Glycation of proteins is the first step in of a nonenzymatic reaction, resulting in advanced glycation end products (AGEs). AGEs can inactivate proteins or modify their biological activities. Therefore, it is important to understand the mechanism of AGE formation. Here, we systematically analyzed the kinetics of AGE formation in vitro by fluorescence and absorption measurements utilizing a microplate reader system and bovine serum albumin (BSA) as a model protein. Comparing different concentrations of BSA, we applied various reducing sugars and reactive dicarbonyl compounds as AGE-inducing agents at different concentrations. In summary, this experimental setup enabled us to measure the kinetics of AGE formation in an efficient and defined way.

  14. Kinetics of advanced glycation end products formation on bovine serum albumin with various reducing sugars and dicarbonyl compounds in equimolar ratios.

    PubMed

    Luers, Lars; Rysiewski, Karolina; Dumpitak, Christian; Birkmann, Eva

    2012-04-01

    Reducing sugars and reactive dicarbonyl compounds play a major role in glycation of proteins in vivo. Glycation of proteins is the first step in of a nonenzymatic reaction, resulting in advanced glycation end products (AGEs). AGEs can inactivate proteins or modify their biological activities. Therefore, it is important to understand the mechanism of AGE formation. Here, we systematically analyzed the kinetics of AGE formation in vitro by fluorescence and absorption measurements utilizing a microplate reader system and bovine serum albumin (BSA) as a model protein. Comparing different concentrations of BSA, we applied various reducing sugars and reactive dicarbonyl compounds as AGE-inducing agents at different concentrations. In summary, this experimental setup enabled us to measure the kinetics of AGE formation in an efficient and defined way. PMID:22533432

  15. Inhibitory effect of 1,2,4-triazole-ciprofloxacin hybrids on Haemophilus parainfluenzae and Haemophilus influenzae biofilm formation in vitro under stationary conditions.

    PubMed

    Kosikowska, Urszula; Andrzejczuk, Sylwia; Plech, Tomasz; Malm, Anna

    2016-10-01

    Haemophilus parainfluenzae and Haemophilus influenzae, upper respiratory tract microbiota representatives, are able to colonize natural and artificial surfaces as biofilm. The aim of the present study was to assay the effect of ten 1,2,4-triazole-ciprofloxacin hybrids on planktonic or biofilm-forming haemophili cells in vitro under stationary conditions on the basis of MICs (minimal inhibitory concentrations) and MBICs (minimal biofilm inhibitory concentrations). In addition, anti-adhesive properties of these compounds were examined. The reference strains of H. parainfluenzae and H. influenzae were included. The broth microdilution microtiter plate (MTP) method with twofold dilution of the compounds, or ciprofloxacin (reference agent) in 96-well polystyrene microplates, was used. The optical density (OD) reading was made spectrophotometrically at a wavelength of 570 nm (OD570) both to measure bacterial growth and to detect biofilm-forming cells under the same conditions with 0.1% crystal violet. The following values of parameters were estimated for 1,2,4-triazole-ciprofloxacin hybrids - MIC = 0.03-15.63 mg/L, MBIC = 0.03-15.63 mg/L, MBIC/MIC = 0.125-8, depending on the compound, and for ciprofloxacin - MIC = 0.03-0.06 mg/L, MBIC = 0.03-0.12 mg/L, MBIC/MIC = 1-2. The observed strong anti-adhesive properties (95-100% inhibition) of the tested compounds were reversible during long-term incubation at subinhibitory concentrations. Thus, 1,2,4-triazole-ciprofloxacin hybrids may be considered as starting compounds for designing improved agents not only against planktonic but also against biofilm-forming Haemophilus spp. cells.

  16. 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).

  17. [Biofilm formation capacity of Listeria monocytogens strains isolated from soft cheese from Costa Rica].

    PubMed

    Carrillo Zeledón, Gabriela; Redondo Solano, Mauricio; Arias Echandi, María Laura

    2010-06-01

    Listeria monocytogenes is a bacteria associated with the production of severe infectious disease in human being, but also with the formation of biofilms in different surfaces related to the food production environment. Biofilm represents a serious problem in food industry, since it is a constant and important contamination source and also, bacteria present in it have an increased resistance towards physical and chemical agents of common use. The capacity of biofilm formation of L. monocytogenes strains previously isolated from soft cheese samples from Costa Rica was studied under different temperature and culture conditions. The microplate technique was performed using different culture media (BHIB, TSB 1:20 and cheese serum) and at different incubation temperatures (refrigeration, environmental and 35 degrees C). Biofilm formation capacity was classified according to the optical density obtained at 620 nm. None of the strains evaluated was classified as strong biofilm former under any of the variables studied, nevertheless, weak and moderate formers were detected. The results obtained show the influence of the nutrient content of the culture media used over biofilm formation; BHIB was the only culture media that allowed the expression of moderate biofilm forms, contrary to cheese serum that did not promote biofilm production. Biofilm formation is a multifactorial process, where adsorption level depends on several variables and its study must be promoted in order to develop methodologies that allow its reduction or elimination, so food industries may offer safe food products to consumers.

  18. 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

  19. 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.

  20. Galaxy formation.

    PubMed

    Peebles, P J

    1998-01-01

    It is argued that within the standard Big Bang cosmological model the bulk of the mass of the luminous parts of the large galaxies likely had been assembled by redshift z approximately 10. Galaxy assembly this early would be difficult to fit in the widely discussed adiabatic cold dark matter model for structure formation, but it could agree with an isocurvature version in which the cold dark matter is the remnant of a massive scalar field frozen (or squeezed) from quantum fluctuations during inflation. The squeezed field fluctuations would be Gaussian with zero mean, and the distribution of the field mass therefore would be the square of a random Gaussian process. This offers a possibly interesting new direction for the numerical exploration of models for cosmic structure formation.

  1. Galaxy formation

    PubMed Central

    Peebles, P. J. E.

    1998-01-01

    It is argued that within the standard Big Bang cosmological model the bulk of the mass of the luminous parts of the large galaxies likely had been assembled by redshift z ∼ 10. Galaxy assembly this early would be difficult to fit in the widely discussed adiabatic cold dark matter model for structure formation, but it could agree with an isocurvature version in which the cold dark matter is the remnant of a massive scalar field frozen (or squeezed) from quantum fluctuations during inflation. The squeezed field fluctuations would be Gaussian with zero mean, and the distribution of the field mass therefore would be the square of a random Gaussian process. This offers a possibly interesting new direction for the numerical exploration of models for cosmic structure formation. PMID:9419326

  2. 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

  3. High Throughput Measurement of Extracellular DNA Release and Quantitative NET Formation in Human Neutrophils In Vitro.

    PubMed

    Sil, Payel; Yoo, Dae-Goon; Floyd, Madison; Gingerich, Aaron; Rada, Balazs

    2016-06-18

    Neutrophil granulocytes are the most abundant leukocytes in the human blood. Neutrophils are the first to arrive at the site of infection. Neutrophils developed several antimicrobial mechanisms including phagocytosis, degranulation and formation of neutrophil extracellular traps (NETs). NETs consist of a DNA scaffold decorated with histones and several granule markers including myeloperoxidase (MPO) and human neutrophil elastase (HNE). NET release is an active process involving characteristic morphological changes of neutrophils leading to expulsion of their DNA into the extracellular space. NETs are essential to fight microbes, but uncontrolled release of NETs has been associated with several disorders. To learn more about the clinical relevance and the mechanism of NET formation, there is a need to have reliable tools capable of NET quantitation. Here three methods are presented that can assess NET release from human neutrophils in vitro. The first one is a high throughput assay to measure extracellular DNA release from human neutrophils using a membrane impermeable DNA-binding dye. In addition, two other methods are described capable of quantitating NET formation by measuring levels of NET-specific MPO-DNA and HNE-DNA complexes. These microplate-based methods in combination provide great tools to efficiently study the mechanism and regulation of NET formation of human neutrophils.

  4. 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

  5. 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.

  6. High Throughput Measurement of Extracellular DNA Release and Quantitative NET Formation in Human Neutrophils In Vitro.

    PubMed

    Sil, Payel; Yoo, Dae-Goon; Floyd, Madison; Gingerich, Aaron; Rada, Balazs

    2016-01-01

    Neutrophil granulocytes are the most abundant leukocytes in the human blood. Neutrophils are the first to arrive at the site of infection. Neutrophils developed several antimicrobial mechanisms including phagocytosis, degranulation and formation of neutrophil extracellular traps (NETs). NETs consist of a DNA scaffold decorated with histones and several granule markers including myeloperoxidase (MPO) and human neutrophil elastase (HNE). NET release is an active process involving characteristic morphological changes of neutrophils leading to expulsion of their DNA into the extracellular space. NETs are essential to fight microbes, but uncontrolled release of NETs has been associated with several disorders. To learn more about the clinical relevance and the mechanism of NET formation, there is a need to have reliable tools capable of NET quantitation. Here three methods are presented that can assess NET release from human neutrophils in vitro. The first one is a high throughput assay to measure extracellular DNA release from human neutrophils using a membrane impermeable DNA-binding dye. In addition, two other methods are described capable of quantitating NET formation by measuring levels of NET-specific MPO-DNA and HNE-DNA complexes. These microplate-based methods in combination provide great tools to efficiently study the mechanism and regulation of NET formation of human neutrophils. PMID:27404503

  7. 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.

  8. Identification of Listeria monocytogenes Determinants Required for Biofilm Formation

    PubMed Central

    Regeimbal, James M.; Regan, Patrick M.; Higgins, Darren E.

    2014-01-01

    Listeria monocytogenes is a Gram-positive, food-borne pathogen of humans and animals. L. monocytogenes is considered to be a potential public health risk by the U.S. Food and Drug Administration (FDA), as this bacterium can easily contaminate ready-to-eat (RTE) foods and cause an invasive, life-threatening disease (listeriosis). Bacteria can adhere and grow on multiple surfaces and persist within biofilms in food processing plants, providing resistance to sanitizers and other antimicrobial agents. While whole genome sequencing has led to the identification of biofilm synthesis gene clusters in many bacterial species, bioinformatics has not identified the biofilm synthesis genes within the L. monocytogenes genome. To identify genes necessary for L. monocytogenes biofilm formation, we performed a transposon mutagenesis library screen using a recently constructed Himar1 mariner transposon. Approximately 10,000 transposon mutants within L. monocytogenes strain 10403S were screened for biofilm formation in 96-well polyvinyl chloride (PVC) microtiter plates with 70 Himar1 insertion mutants identified that produced significantly less biofilms. DNA sequencing of the transposon insertion sites within the isolated mutants revealed transposon insertions within 38 distinct genetic loci. The identification of mutants bearing insertions within several flagellar motility genes previously known to be required for the initial stages of biofilm formation validated the ability of the mutagenesis screen to identify L. monocytogenes biofilm-defective mutants. Two newly identified genetic loci, dltABCD and phoPR, were selected for deletion analysis and both ΔdltABCD and ΔphoPR bacterial strains displayed biofilm formation defects in the PVC microtiter plate assay, confirming these loci contribute to biofilm formation by L. monocytogenes. PMID:25517120

  9. 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

  10. 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.

  11. A novel, generic, electroanalytical immunoassay format utilising silver nano-particles as a bio-label.

    PubMed

    Porter, Robert; Kabil, Alena; Forstern, Camilla; Slevin, Christopher; Kouwenberg, Katherine; Szymanski, Mateusz; Birch, Brian

    2009-01-01

    The aim of this study was to evaluate a novel, generic, sensitive electroanalytical platform method for binding reactions, such as immuno or DNA assays. It was thought that silver nano-particles of 40 nM when attached to the analyte of interest would give an electroanalytical amplification of approx. 10(6) resulting from silver ions produced by dissolution of the nano-particles by a mild chemical oxidant such as ferricyanide. Ferricyanide has been widely used in biochemical measurement and has been shown to relatively stable in a biosensor device. Here we have demonstrated the use of silver nano-particles as a bio-conjugate in a 96 well sandwich assay format for measuring human chorionic gonadotropin (hCG) to a concentration of 0.2 mIU.

  12. 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.

  13. 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

  14. 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

  15. Analyzing abundance of mRNA molecules with a near-infrared fluorescence technique.

    PubMed

    Chen, Ying; Pan, Yan; Zhang, Beibei; Wang, Jinke

    2014-01-01

    This study describes a simple method for analyzing the abundance of mRNA molecules in a total DNA sample. Due to the dependence on the near-infrared fluorescence technique, this method is named near-infrared fluorescence gene expression detection (NIRF-GED). The procedure has three steps: (1) isolating total RNA from detected samples and reverse-transcription into cDNA with a biotin-labeled oligo dT; (2) hybridizing cDNA to oligonucleotide probes coupled to a 96-well microplate; and (3) detecting biotins with NIRF-labeled streptavidin. The method was evaluated by performing proof-in-concept detections of absolute and relative expressions of housekeeping and NF-κB target genes in HeLa cells. As a result, the absolute expression of three genes, Ccl20, Cxcl2, and Gapdh, in TNF-α-uninduced HeLa cells was determined with a standard curve constructed on the same microplate, and the relative expression of five genes, Ccl20, Cxcl2, Il-6, STAT5A, and Gapdh, in TNF-α-induced and -uninduced HeLa cells was measured by using NIRF-GED. The results were verified by quantitative PCR (qPCR) and DNA microarray detections. The biggest advantage of NIRF-GED over the current techniques lies in its independence of exponential or linear amplification of nucleic acids. Moreover, NIRF-GED also has several other benefits, including high sensitivity as low as several fmols, absolute quantification in the range of 9 to 147 fmols, low cDNA consumption similar to qPCR template, and the current medium throughput in 96-well microplate format and future high throughput in DNA microarray format. NIRF-GED thus provides a new tool for analyzing gene transcripts and other nucleic acid molecules. PMID:24317515

  16. The natural antimicrobial carvacrol inhibits quorum sensing in Chromobacterium violaceum and reduces bacterial biofilm formation at sub-lethal concentrations.

    PubMed

    Burt, Sara A; Ojo-Fakunle, Victoria T A; Woertman, Jenifer; Veldhuizen, Edwin J A

    2014-01-01

    The formation of biofilm by bacteria confers resistance to biocides and presents problems in medical and veterinary clinical settings. Here we report the effect of carvacrol, one of the major antimicrobial components of oregano oil, on the formation of biofilms and its activity on existing biofilms. Assays were carried out in polystyrene microplates to observe (a) the effect of 0-0.8 mM carvacrol on the formation of biofilms by selected bacterial pathogens over 24 h and (b) the effect of 0-8 mM carvacrol on the stability of pre-formed biofilms. Carvacrol was able to inhibit the formation of biofilms of Chromobacterium violaceum ATCC 12472, Salmonella enterica subsp. Typhimurium DT104, and Staphylococcus aureus 0074, while it showed no effect on formation of Pseudomonas aeruginosa (field isolate) biofilms. This inhibitory effect of carvacrol was observed at sub-lethal concentrations (<0.5 mM) where no effect was seen on total bacterial numbers, indicating that carvacrol's bactericidal effect was not causing the observed inhibition of biofilm formation. In contrast, carvacrol had (up to 8 mM) very little or no activity against existing biofilms of the bacteria described, showing that formation of the biofilm also confers protection against this compound. Since quorum sensing is an essential part of biofilm formation, the effect of carvacrol on quorum sensing of C. violaceum was also studied. Sub-MIC concentrations of carvacrol reduced expression of cviI (a gene coding for the N-acyl-L-homoserine lactone synthase), production of violacein (pigmentation) and chitinase activity (both regulated by quorum sensing) at concentrations coinciding with carvacrol's inhibiting effect on biofilm formation. These results indicate that carvacrol's activity in inhibition of biofilm formation may be related to the disruption of quorum sensing.

  17. Effects of nutritional and environmental conditions on planktonic growth and biofilm formation of Citrobacter werkmanii BF-6.

    PubMed

    Zhou, Gang; Li, Long-jie; Shi, Qing-shan; Ouyang, You-sheng; Chen, Yi-ben; Hu, Wen-feng

    2013-12-01

    Citrobacter sp. is a cause of significant opportunistic nosocomial infection and is frequently found in human and animal feces, soil, and sewage water, and even in industrial waste or putrefaction. Biofilm formation is an important virulence trait of Citrobacter sp. pathogens but the process and characteristics of this formation are unclear. Therefore, we employed in vitro assays to study the nutritional and environmental parameters that might influence biofilm formation of C. werkmanii BF-6 using 96-well microtiter plates. In addition, we detected the relative transcript levels of biofilm formation genes by RT-PCR. Our results indicated that the capacity of C. werkmanii BF-6 to form biofilms was affected by culture temperature, media, time, pH, and the osmotic agents glucose, sucrose, NaCl, and KCl. Confocal laser scanning microscopy results illustrated that the structure of biofilms and extracellular polysaccharide was influenced by 100 mM NaCl or 100 mM KCl. In addition, nine biofilm formation genes (bsmA, bssR, bssS, csgD, csgE, csgF, mrkA, mrkB, and mrkE) were found to contribute to planktonic and biofilm growth. Our data suggest that biofilm formation by C. werkmanii BF-6 is affected by nutritional and environmental factors, which could pave the way to the prevention and elimination of biofilm formation using proper strategies.

  18. 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

  19. Motility of Pseudomonas aeruginosa contributes to SOS-inducible biofilm formation.

    PubMed

    Chellappa, Shakinah T; Maredia, Reshma; Phipps, Kara; Haskins, William E; Weitao, Tao

    2013-12-01

    DNA-damaging antibiotics such as ciprofloxacin induce biofilm formation and the SOS response through autocleavage of SOS-repressor LexA in Pseudomonas aeruginosa. However, the biofilm-SOS connection remains poorly understood. It was investigated with 96-well and lipid biofilm assays. The effects of ciprofloxacin were examined on biofilm stimulation of the SOS mutant and wild-type strains. The stimulation observed in the wild-type in which SOS was induced was reduced in the mutant in which LexA was made non-cleavable (LexAN) and thus SOS non-inducible. Therefore, the stimulation appeared to involve SOS. The possible mechanisms of inducible biofilm formation were explored by subproteomic analysis of outer membrane fractions extracted from biofilms. The data predicted an inhibitory role of LexA in flagellum function. This premise was tested first by functional and morphological analyses of flagellum-based motility. The flagellum swimming motility decreased in the LexAN strain treated with ciprofloxacin. Second, the motility-biofilm assay was performed, which tested cell migration and biofilm formation. The results showed that wild-type biofilm increased significantly over the LexAN. These results suggest that LexA repression of motility, which is the initial event in biofilm development, contributes to repression of SOS-inducible biofilm formation.

  20. High-throughput flow injection analysis mass spectroscopy with networked delivery of color-rendered results. 2. Three-dimensional spectral mapping of 96-well combinatorial chemistry racks.

    PubMed

    Görlach, E; Richmond, R; Lewis, I

    1998-08-01

    For the last two years, the mass spectroscopy section of the Novartis Pharma Research Core Technology group has analyzed tens of thousands of multiple parallel synthesis samples from the Novartis Pharma Combinatorial Chemistry program, using an in-house developed automated high-throughput flow injection analysis electrospray ionization mass spectroscopy system. The electrospray spectra of these samples reflect the many structures present after the cleavage step from the solid support. The overall success of the sequential synthesis is mirrored in the purity of the expected end product, but the partial success of individual synthesis steps is evident in the impurities in the mass spectrum. However this latter reaction information, which is of considerable utility to the combinatorial chemist, is effectively hidden from view by the very large number of analyzed samples. This information is now revealed at the workbench of the combinatorial chemist by a novel three-dimensional display of each rack's complete mass spectral ion current using the in-house RackViewer Visual Basic application. Colorization of "forbidden loss" and "forbidden gas-adduct" zones, normalization to expected monoisotopic molecular weight, colorization of ionization intensity, and sorting by row or column were used in combination to highlight systematic patterns in the mass spectroscopy data.

  1. 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 ...

  2. Cretaceous evolution of the Indian Plate and consequences for the formation, deformation and obduction of adjacent oceanic crust

    NASA Astrophysics Data System (ADS)

    Gaina, C.; Van Hinsbergen, D. J.; Spakman, W.

    2012-12-01

    As part of the gradual Gondwana dispersion that started in the Jurassic, the Indian tectonic block was rifted away from the Antarctica-Australian margins, probably in the Early-Mid Cretaceous and started its long journey to the north until it collided with Eurasia in the Tertiary. In this contribution first we will revise geophysical and geological evidences for the formation of oceanic crust between India and Antarctica, India and Madagascar, and India and Somali/Arabian margins. This information and possible oceanic basin age interpretation are placed into regional kinematic models. Three important compressional events NW and W of the Indian plate are the result of the opening of the Enderby Basin from 132 to 124 Ma, the first phase of seafloor spreading in the Mascarene basin approximately from 84 to 80 Ma, and the incipient opening of the Arabian Sea and the Seychelles microplate formation around 65 to 60 Ma. Based on retrodeformation of the Afghan-Pakistan part of the India-Asia collision zone and the eastern Oman margin, the ages of regional ophiolite emplacement and crystallization of its oceanic crust, as well as the plate tectonic setting of these ophiolites inferred from its geochemistry, we evaluate possible scenarios for the formation of intra-oceanic subduction zones and their evolution until ophiolite emplacement time. Our kinematic scenarios are constructed for several regional models and are discussed in the light of global tomographic models that may image some of the subducted Cretaceous oceanic lithosphere.

  3. [Biofilm formation by different strains of Salmonella typhimurium in artificial systems].

    PubMed

    Romanova, Iu M; Alekseeva, N V; Smirnova, T A; Andreev, A L; Didenko, L V; Gintsburg, A L

    2006-01-01

    The ability of 14 different strains of Salmonella typhimurium to biofilm formation depending on genotype and culture conditions was investigated in artificial systems: in 96-well plastic microtitre plates, plastic and glass tubes, plastic Petri dishes and on microscope glasses. Quantitative biofilm growth was monitored by using an assay based on crystal violet staining, while planctonic growth in the same cultures was monitored by absorbance in iEMS Reader MF, and qualitatively--by digital photo and visually. Optimal rate between growth and biofilm indications for all strains was determined at initial cell concentration 10(6-7) KOE/ml and culture incubation at t degrees 28 degrees C. The nutrient content of the medium significantly influenced the quantity of produced biofilm. The nutrient broth LB without NaCl was more effective in promoting biofilm formation, than LB itself. The least quantity of biofilm was formed in water. The genotype of the strains also critically influenced the quantity of produced biofilm. Nonmotile mutants cells had reduced ability to form biofilm. RpoS mutant cells produced significantly less biofilm as compared with cells of isogenic parent strains. The chemical content of plastic and glass also influenced biofilm formation.

  4. Nitric Oxide Regulates Neutrophil Migration through Microparticle Formation

    PubMed Central

    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 NG-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

  5. Common File Formats.

    PubMed

    Mills, Lauren

    2014-03-21

    An overview of the many file formats commonly used in bioinformatics and genome sequence analysis is presented, including various data file formats, alignment file formats, and annotation file formats. Example workflows illustrate how some of the different file types are typically used.

  6. 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

  7. Inhibition on Candida albicans biofilm formation using divalent cation chelators (EDTA).

    PubMed

    Ramage, Gordon; Wickes, Brian L; López-Ribot, José L

    2007-12-01

    Candida albicans can readily form biofilms on both inanimate and biological surfaces. In this study we investigated a means of inhibiting biofilm formation using EDTA (Ethylenediaminetetra-acetic acid), a divalent cation chelating agent, which has been shown to affect C. albicans filamentation. Candida albicans biofilms were formed in 96-well microtitre plates. Cells were allowed to adhere for 1, 2, and 4 h at 37 degrees C, washed in PBS, and then treated with different concentrations of EDTA (0, 2.5, 25, and 250 mM). EDTA was also added to the standardized suspension prior to adding to the microtiter plate and to a preformed 24 h biofilm. All plates were then incubated at 37 degrees C for an additional 24 h to allow for biofilm formation. The extent and characteristics of biofilm formation were then microscopically assessed and with a semi-quantitative colorimetric technique based on the use of an XTT-reduction assay. Northern blot analysis of the hyphal wall protein (HWP1) expression was also monitored in planktonic and biofilm cells treated with EDTA. Microscopic analysis and colorimetric readings revealed that filamentation and biofilm formation were inhibited by EDTA in a concentration dependent manner. However, preformed biofilms were minimally affected by EDTA (maximum of 31% reduction at 250 mM). The HWP1 gene expression was reduced in EDTA-treated planktonic and biofilm samples. These results indicate that EDTA inhibits C. albicans biofilm formation are most likely through its inhibitory effect on filamentation and indicates the potential therapeutic effects of EDTA. This compound may serve a non-toxic means of preventing biofilm formation on infections with a C. albicans biofilm etiology. PMID:17909983

  8. 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

  9. 2(5H)-Furanone, epigallocatechin gallate, and a citric-based disinfectant disturb quorum-sensing activity and reduce motility and biofilm formation of Campylobacter jejuni.

    PubMed

    Castillo, Sandra; Heredia, Norma; García, Santos

    2015-01-01

    Brominated furanone and epigallocatechin gallate (EGCG) are naturally occurring polyphenolic compounds that can be derived from sources such as Delisea pulchra algae and green tea, respectively. These compounds may have potential health benefits and antimicrobial properties. Biofilm formation and bacterial motility are virulence factors that seem to be involved in the autoinducer 2 (AI-2)-mediated quorum sensing (QS) response of Campylobacter. In this study, the anti-QS activities of 2(5H)-furanone, EGCG, and a citric-based disinfectant were tested against Campylobacter jejuni. The minimal bactericidal concentration (MBC) was determined by a microdilution method, and the AI-2 activity was measured by bioluminescence. For motility tests, subinhibitory concentrations of each compound were mixed with semisolid Muller Hinton agar. Biofilm formation was quantified in broth-containing microplates after staining with safranin. The MBC of tested compounds ranged from 0.3 to 310 μg/mL. Subinhibitory concentrations of all of the antimicrobial compounds significantly decreased (19 to 62 %) the bacterial motility and reduced biofilm formation. After treatment with EGCG, furanone, and the disinfectant, AI-2 activity was decreased by 60 to 99 % compared to control. In conclusion, 2(5H)-furanone, EGCG, and the disinfectant exert bactericidal effects against C. jejuni and disturb QS activity and reduce motility and biofilm formation. These compounds may be naturally occurring alternatives to control C. jejuni.

  10. 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

  11. 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)

  12. 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.

  13. 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.

  14. Record Formatting: MARC AMC.

    ERIC Educational Resources Information Center

    Weber, Lisa B.

    1990-01-01

    Discusses how archivists use the MARC AMC (Archival and Manuscripts Control) format for descriptive information about archival materials. Modifications that MARC AMC introduced to the standard MARC bibliographic formats are explained, and examples of a record in AMC formats on different bibliographic utilities are given. (24 references) (LRW)

  15. 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.

  16. 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.

  17. 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

  18. 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.

  19. 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.

  20. 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

  1. Inhibition of Biofilm Formation, Quorum Sensing and Infection in Pseudomonas aeruginosa by Natural Products-Inspired Organosulfur Compounds

    PubMed Central

    Cady, Nathaniel C.; McKean, Kurt A.; Behnke, Jason; Kubec, Roman; Mosier, Aaron P.; Kasper, Stephen H.; Burz, David S.; Musah, Rabi A.

    2012-01-01

    Using a microplate-based screening assay, the effects on Pseudomonas aeruginosa PAO1 biofilm formation of several S-substituted cysteine sulfoxides and their corresponding disulfide derivatives were evaluated. From our library of compounds, S-phenyl-L-cysteine sulfoxide and its breakdown product, diphenyl disulfide, significantly reduced the amount of biofilm formation by P. aeruginosa at levels equivalent to the active concentration of 4-nitropyridine-N-oxide (NPO) (1 mM). Unlike NPO, which is an established inhibitor of bacterial biofilms, our active compounds did not reduce planktonic cell growth and only affected biofilm formation. When used in a Drosophila-based infection model, both S-phenyl-L-cysteine sulfoxide and diphenyl disulfide significantly reduced the P. aeruginosa recovered 18 h post infection (relative to the control), and were non-lethal to the fly hosts. The possibility that the observed biofilm inhibitory effects were related to quorum sensing inhibition (QSI) was investigated using Escherichia coli-based reporters expressing P. aeruginosa lasR or rhIR response proteins, as well as an endogenous P. aeruginosa reporter from the lasI/lasR QS system. Inhibition of quorum sensing by S-phenyl-L-cysteine sulfoxide was observed in all of the reporter systems tested, whereas diphenyl disulfide did not exhibit QSI in either of the E. coli reporters, and showed very limited inhibition in the P. aeruginosa reporter. Since both compounds inhibit biofilm formation but do not show similar QSI activity, it is concluded that they may be functioning by different pathways. The hypothesis that biofilm inhibition by the two active compounds discovered in this work occurs through QSI is discussed. PMID:22715388

  2. 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.

  3. 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.

  4. [Measuring thrombin formation].

    PubMed

    Hemker, H C

    2016-01-01

    Measurement of thrombin formation makes it possible to estimate the risk of haemorrhage or thrombosis much more accurately than by using clotting time. This new technique allows better monitoring of the effect of prophylactic and therapeutic anticoagulant therapy. Thrombin formation is, however, not yet routinely measured. PMID:27650017

  5. 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.

  6. 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…

  7. Entering the Formative Years

    ERIC Educational Resources Information Center

    Powers, Keith

    2011-01-01

    This article discusses formative assessment--classroom strategies that ensure students are understanding music concepts. Unlike summative assessments (end-of-process evaluations like final exams, SATs, or auditions), formative assessments need to be non-threatening, helpful, and most of all, effective. The process starts with a teacher…

  8. 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…

  9. Rapid LC-MS drug metabolite profiling using microsomal enzyme bioreactors in a parallel processing format.

    PubMed

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

    2009-12-15

    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

  10. Intergalactic Star Formation

    NASA Astrophysics Data System (ADS)

    Boquien, Médéric

    2007-11-01

    The work presented here is about star formation in the unusual environment of collisional debris studied for the first time as such. These peculiar regions have an interstellar medium, and in particular a metallicity, similar to that of star forming regions in galactic discs while not undergoing similar environment effects such as density waves in the spiral arms for instance. This study has been conducted with a selection of exceptional systems that have ejected large quantities of gas into the intergalactic medium while also showing some intergalactic star forming regions. Principal Investigator as well as archive spectroscopy and imaging from multi-wavelength observations ranging from far ultraviolet to mid-infrared have been used. Withal a model has been built in order to reproduce the spectral energy distributions of intergalactic star forming regions and constrain the star formation histories, their extinctions and their fraction of stars coming from the parent galaxies' discs. Comparisons have been performed on the estimation of star formation rates between infrared, Halpha and ultraviolet wavelengths. This thesis has brought the following main new results: * some regions seem to be deprived of any old stellar population, and these are ideal laboratories in which to study star formation ; * the mid-infrared star formation rate estimator is as reliable as it is in spiral galaxies ; * the scatter in the estimation of star formation rates in various bands is similar to that of spiral galaxies and is mainly due to age effects ; * the combination of the extinction uncorrected Halpha line with mid-infrared yields a good estimation of the actual star formation rate ; * an important part of star formation, which can be as high as 85%, takes place in the intergalactic medium showing that in a young universe, in which this type of system is much more common than in the nearby universe, star formation from collisional debris can be an important factor of enrichment of

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. Biofilm Formation and Susceptibility to Amphotericin B and Fluconazole in Candida albicans

    PubMed Central

    Zarei Mahmoudabadi, Ali; Zarrin, Majid; Kiasat, Neda

    2014-01-01

    Background: The ability of Candida albicans to form biofilms and adhere to host tissues and biomaterial surfaces is an important factor in its pathogenesis. One of the main characteristics of biofilms is their resistance to broad-spectrum anti-microbial drugs. Objectives: In the present study the formation of biofilm by C. albicans from different sources was evaluated. In addition, the minimum biofilm inhibitory concentration (MBIC) for two antifungals was evaluated. Materials and Methods: In total, 120 isolates of C. albicans from different sources (patients with vaginitis, patients with candiduria, bucal cavity and environmental surfaces) were collected. Biofilm formation was determined by the 96-well micro-titeration plate method. MBIC testing was also performed, using the calorimetric indicator resazurin for amphotericin B and fluconazole. Results: The results indicated that 100% of C. albicans isolates from different sources had the ability to form biofilms in vitro. Amongst these isolates, 83.3% of isolates had the maximum potential (4+) to form biofilms, while only one (0.9%) of isolates had the minimum ability (1+) to form biofilms. Our results showed that 65.0% of the tested isolates are sensitive to amphotericin B at amounts lower than 10 µg/mL, while only 26.7% are sensitive to fluconazole (had MBIC < 10 µg/mL). Conclusions: Although biofilm formation was detected in all tested isolates, there were differences in the ability to form biofilms between isolates from different sources. In addition, there were differences in the MBIC against the two examined antifungals, amphotericin B and fluconazole. PMID:25368806

  16. 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 ...

  17. 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, ...

  18. 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.

  19. 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.

  20. 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.

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

    PubMed Central

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

    2013-01-01

    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

  2. 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

  3. 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

  4. 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

  5. Planet Formation and Habitability

    NASA Astrophysics Data System (ADS)

    alibert, yann

    2016-04-01

    Extrasolar planetary systems show an extreme diversity in mass and orbital architecture, and, very likely, in habitability. Explaining this diversity is one of the key challenges for theoretical models and requires understanding the formation, composition and evolution of planetary systems from the stage of the protoplanetary disk up to the full mature planetary system. I will review in this contribution the different models of planet formation and how they can be related to planetary habitability. In a first part, I will review the main planetary system formation models, and how, from these models, the composition of planets can be predicted. In a second part, I will link the results of these early phases of planetary systems, to the potential planetary habitability. Finally, I will show how it is possible, from transit observations, to put constraints on the water content of extrasolar planets.

  6. 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.

  7. 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

  8. 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

  9. 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)

  10. 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.

  11. Carrascolendas: A Formative Evaluation.

    ERIC Educational Resources Information Center

    Laosa, Luis M.

    A formative research project sought to test viewer reactions to two pilot programs of the Carrascolendas series. A total of 360 Puerto Rican-American, Cuban-American, Mexican-American, and Anglo-American children in grades 1, 2, and 3 were observed as they watched the programs. Results indicated that there was high eye contact during the…

  12. 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…

  13. 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…

  14. Concept Formation and Abstraction.

    ERIC Educational Resources Information Center

    Lunzer, Eric A.

    1979-01-01

    This paper examines the nature of concepts and conceptual processes and the manner of their formation. It argues that a process of successive abstraction and systematization is central to the evolution of conceptual structures. Classificatory processes are discussed and three levels of abstraction outlined. (Author/SJL)

  15. 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

  16. 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.

  17. Generic file format

    NASA Astrophysics Data System (ADS)

    Felgate, Nick

    2002-11-01

    The Generic File Format (GFF) is a file format developed within the UK ASW community for the interchange and storage of underwater sonar data. Originally developed for the interchange of time-series data between analysis systems, it has been extended to provide for storage of processed acoustic data (e.g., power and DEMON spectrum, lofargram grey-scale), nonacoustic data (e.g., own-ship dynamics, sensor configuration) and event data (e.g., tracker output, sonar intercepts). The format employs the chunk concept, as used in the WAV and AIFF file formats, to provide extendability (including local variants) while providing a measure of backward compatability. However, the basic concept has been adapted to allow for the mixing in the one file of multiple channels of different sample-rates and data-types through the inclusion of a data frame concept and multiple data blocks. Chunk cross-referencing has been employed to ensure data consistency. A provision is made in the header of the file to store details of the sensor and processing for the data (e.g., the number of hydrophones, beam direction, FFT size) so that an analysis system does not need to know about the sensor or other system from which the data originated.

  18. 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)

  19. 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

  20. High-throughput screening with HyperCyt flow cytometry to detect small molecule formylpeptide receptor ligands.

    PubMed

    Young, Susan M; Bologa, Cristian; Prossnitz, Eric R; Oprea, Tudor I; Sklar, Larry A; Edwards, Bruce S

    2005-06-01

    High-throughput flow cytometry (HTFC), enabled by faster automated sample processing, represents a promising high- content approach for compound library screening. HyperCyt is a recently developed automated HTFC analysis system by which cell samples are rapidly aspirated from microplate wells and delivered to the flow cytometer. The formylpeptide receptor (FPR) family of G protein-coupled receptors contributes to the localization and activation of tissue-damaging leukocytes at sites of chronic inflammation. Here, the authors describe development and application of an HTFC screening approach to detect potential anti-inflammatory compounds that block ligand binding to FPR. Using a homogeneous no-wash assay, samples were routinely processed at 1.5 s/well (approximately 2500 cells analyzed/sample), allowing a 96-well plate to be processed in less than 2.5 min. Assay sensitivity and accuracy were validated by detection of a previously documented active compound with relatively low FPR affinity (sulfinpyrazone, inhibition constant [K(i)]=14 microM) from among a collection of 880 compounds in the Prestwick Chemical Library. The HyperCyt system was therefore demonstrated to be a robust, sensitive, and highly quantitative method with which to screen lead compound libraries in a 96-well format. PMID:15964939

  1. Resistance to antimicrobials and biofilm formation in Staphylococcus spp. isolated from bovine mastitis in the Northeast of Brazil.

    PubMed

    da Costa Krewer, Carina; Santos Amanso, Evandro; Veneroni Gouveia, Gisele; de Lima Souza, Renata; da Costa, Mateus Matiuzzi; Aparecido Mota, Rinaldo

    2015-03-01

    Mastitis is the principal disease affecting dairy herds worldwide. The aim of the present study was to characterize phenotypic and genotypic features associated with resistance to antimicrobials in Staphylococcus spp. isolated from 2064 milk samples of 525 lactating cows in the Northeast of Brazil. Of the 218 isolates analyzed, 57.8% were characterized as Staphylococcus aureus, 28% as coagulase-positive staphylococci other than S. aureus (oCPS), and 14.2% as coagulase-negative staphylococci (CNS). The test for susceptibility to antimicrobials showed amoxicillin (32.6%) to be the less effective drug in vitro, and the multi-drug resistance (MDR) rate for beta-lactams varied from 0 to 0.75. The genotypic characterization showed that 93.1% of the samples were tested positive for the blaZ gene, while none amplified mecA. The antibiotic efflux mechanism was observed in 0.9% of isolates. The biofilm formation was found in 3.7 and 96.3% of samples, respectively, on Congo red agar and on the microplate adhesion test, while the icaD gene was present in 92.2% of Staphylococcus spp. The high frequency of blaZ gene observed in this study was associated with the resistance of most Staphylococcus spp. to one or more of the beta-lactams tested, which are routinely used in Brazilian herds for mastitis treatment. The biofilm formation was also detected in the isolates analyzed being an important characteristic for pathogenicity and antimicrobial resistance of bacteria.

  2. 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

  3. 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

  4. 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).

  5. 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.

  6. Tetrahedron Formation Control

    NASA Technical Reports Server (NTRS)

    Petruzzo, Charles; Guzman, Jose

    2004-01-01

    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 (1) targeting to a fixed tetrahedron location and orientation, and (2) rotating and translating the tetrahedron. The number of impulsive maneuvers can 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 NASA Goddard Magnetospheric Multi-Scale (MMS) mission to compute preliminary formation control fuel requirements.

  7. 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.

  8. 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.

  9. 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.

  10. Simulations of antihydrogen formation

    SciTech Connect

    Robicheaux, F.

    2004-08-01

    The results of simulations of antihydrogen formation in a Penning trap are reported. The antihydrogen atoms are formed by three-body capture. We find that the arrested nature of the three-body capture in the trap greatly reduces the expected binding energy of the antihydrogen. Typically, the formed antihydrogen has larger velocity along the magnetic field than across the field and a binding energy below k{sub B}T.

  11. Formate-assisted pyrolysis

    DOEpatents

    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.

  12. 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.

  13. 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

  14. 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.

  15. 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.

  16. 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

  17. Mars brine formation experiment

    NASA Astrophysics Data System (ADS)

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

    1993-09-01

    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 (106 - 107 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.

  18. 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

  19. Extracts of three Laserpitium L. species and their principal components laserpitine and sesquiterpene lactones inhibit microbial growth and biofilm formation by oral Candida isolates.

    PubMed

    Popović, Višnja; Stojković, Dejan; Nikolić, Miloš; Heyerick, Arne; Petrović, Silvana; Soković, Marina; Niketić, Marjan

    2015-04-01

    Antimicrobial properties of extracts of underground parts of three Laserpitium L. (Apiaceae) species, namely Laserpitium latifolium L., Laserpitium zernyi Hayek and Laserpitium ochridanum Micevski, were investigated. The investigated species are widely used as functional foods, as spices and for preparations in traditional medicine for treating complaints connected with infection and inflammation. Furthermore, antimicrobial and antibiofilm effects of laserpitine, the most abundant compound in the chloroform extract of L. latifolium, and guaianolide sesquiterpene lactones, such as, isomontanolide, montanolide and tarolide, principal components of the extracts of L. zernyi and L. ochridanum were assessed. The antimicrobial activity was tested using the microdilution method against five pathogenic bacteria and five fungi, as well as in the microplate biofilm assay on two Candida clinical isolates (C. albicans and C. krusei). Among the extracts, L. latifolium showed the most prominent activity. Isolated metabolites exerted higher effects against fungal than against bacterial strains, isomontanolide being the most active. Interestingly, all constituents showed higher potential on inhibition of biofilm formation than fluconazole, a reference compound. Tested metabolites may be good novel agents with high antifungal and antibacterial potential that might find practical applications in food industry as food preservatives in order to retard the growth of food spoiling microbes, but only after detailed safety assessments. PMID:25720441

  20. Screening and classifying small-molecule inhibitors of amyloid formation using ion mobility spectrometry-mass spectrometry.

    PubMed

    Young, Lydia M; Saunders, Janet C; Mahood, Rachel A; Revill, Charlotte H; Foster, Richard J; Tu, Ling-Hsien; Raleigh, Daniel P; Radford, Sheena E; Ashcroft, Alison E

    2015-01-01

    The search for therapeutic agents that bind specifically to precursor protein conformations and inhibit amyloid assembly is an important challenge. Identifying such inhibitors is difficult because many protein precursors of aggregation are partially folded or intrinsically disordered, which rules out structure-based design. Furthermore, inhibitors can act by a variety of mechanisms, including specific or nonspecific binding, as well as colloidal inhibition. Here we report a high-throughput method based on ion mobility spectrometry-mass spectrometry (IMS-MS) that is capable of rapidly detecting small molecules that bind to amyloid precursors, identifying the interacting protein species and defining the mode of inhibition. Using this method we have classified a variety of small molecules that are potential inhibitors of human islet amyloid polypeptide (hIAPP) aggregation or amyloid-beta 1-40 aggregation as specific, nonspecific, colloidal or non-interacting. We also demonstrate the ability of IMS-MS to screen for inhibitory small molecules in a 96-well plate format and use this to discover a new inhibitor of hIAPP amyloid assembly.

  1. 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

  2. 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.

  3. What initiated planetesimal formation?

    NASA Technical Reports Server (NTRS)

    Cuzzi, J. N.; Dobrovolskis, A. R.; Hogan, R. C.

    1994-01-01

    The physical structure of primitive (chondritic) meteorites, even after some geological processing and modification, is thought by most to contain clues as to the first stage of accretion of solid matter into objects that might be called planetesimals. However, theoretical understanding of the processes responsible for this important stage is shaky. We note what we believe are fundamental obstacles for the Goldreich-Ward version of rapid and direct planetesimal formation via gravitational instability in a settled particle layer, and describe an alternative scenario which might lead from grainy nebula gas to primitive planetesimals in a way that has intriguing connections to the meteorite evidence.

  4. 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

  5. Ultrarelativistic black hole formation.

    PubMed

    East, William E; Pretorius, Frans

    2013-03-01

    We study the head-on collision of fluid particles well within the kinetic energy dominated regime (γ = 8 to 12) by numerically solving the Einstein-hydrodynamic equations. We find that the threshold for black hole formation is lower (by a factor of a few) than simple hoop conjecture estimates, and, moreover, near this threshold two distinct apparent horizons first form postcollision and then merge. We argue that this can be understood in terms of a gravitational focusing effect. The gravitational radiation reaches luminosities of 0.014 c(5)/G, carrying 16 ± 2% of the total energy.

  6. 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.

  7. Flexible formation configuration for terrain following flight: Formation keeping constraints

    NASA Astrophysics Data System (ADS)

    Latyshev, Simon

    This work suggests a control method for the terrain-following formation motion of a group of communicating autonomous agents. The presented approach centers on defining a suitable set of constraints for formation keeping task that shall be fulfilled while agents are negotiating an unknown terrain toward the predefined goal location. It allows agents to maintain a general geometric formation shape, while allowing each individual formation member freedom of maneuver, required for terrain collision free motion. Formation structure is defined with the use of virtual leader. Formation keeping constraints are defined with plane surfaces, specified relative to position and navigation vector of the virtual leader. Formation navigation and guidance constraints are defined using navigation vectors of formation members and the virtual leader. Alternative designs for the constraints derived with parabolic, cone, and cylindrical surfaces are considered. Formation control is derived using the Udwadia-Kalaba equation, following corresponding approach to the development of control methods for constraint based dynamical systems, including leader-follower systems defined using geometric constraints. Approach to terrain following motion requiring agents to stay within bounds of cylindrical corridor volumes built around their respective navigation vectors is assumed. Individual formation primitives and multi-level, hierarchical, formation structures are considered. Simulations, based on three degrees of freedom nonlinear model of an agent, performed using Mathematica and specifically developed combined Maya-Mathematica modeling and simulation system, demonstrate that a flexible terrain following formation motion is achieved with the presented sets of constraints.

  8. 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)

  9. Standard exercise report format (SERF)

    SciTech Connect

    1995-12-31

    This talk summarizes the reasons for the development of draft SERF the Standard Exercise Report Format used for reporting the results of emergency preparedness exercises, and gives a summary of the format and rational behind it.

  10. 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)

  11. Tooth formation - delayed or absent

    MedlinePlus

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

  12. 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.

  13. 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.

  14. 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.

  15. 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…

  16. Statins and bone formation.

    PubMed

    Garrett, I R; Gutierrez, G; Mundy, G R

    2001-05-01

    The main therapy needed most in the bone field is an anabolic agent for the treatment of osteoporosis. Current drugs on the market, which included bisphosphonates, calcitonin, estrogen and related compounds, vitamin D analogues trabecular microarchitecture. Therefore, it would be desirable to have a satisfactory and universally and iprifalvone, are essentially bone resorption inhibitors that mainly act to stabilize bone mass. Patients with established osteoporosis have lost more than 50% of their bone mass at critical sites in the skeleton, and more over have marked disruption of acceptable drug that would stimulate new bone formation and correct this disturbance of trabecular microarchitecture characteristic of established osteoporosis. Recently inhibitors of the enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, which controls the first step in the biosynthesis of cholesterol, have been shown to stimulate bone formation in rodents both in vitro and in vivo. The effect is associated with an increased expression of the bone morphogenetic protein-2 (BMP-2) gene in bone cells. These statins drugs are widely used agents for lowering cholesterol and reducing heart attacks, however they are also known to elicit numerous pleiotropic effects including inhibition of proliferation and migration of smooth muscle cells, inhibition of tumor growth and anti-inflammatory activity. Some of these effects have been attributed to not only to the reduction of cholesterol synthesis by inhibition of the HMG-CoA reductase enzyme but also by the concurrent reduction in downstream metabolites of the mevalonate pathway such as mevalonate, farnesyl pyrophosphate and geranylgeranyl pyrophosphate. The findings that statins are capable of increasing bone formation and bone mass in rodents suggests a potential new action for the statins, which may be beneficial in patients with established osteoporosis where marked bone loss has occurred. Recent clinical data suggests that they

  17. 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.

  18. 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

  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. Transitions in biofilm formation

    NASA Astrophysics Data System (ADS)

    Gordon, Vernita; Thatcher, Travis; Cooley, Benjamin

    2011-03-01

    Biofilms are multicellular, dynamic communities formed by interacting unicellular organisms bound to a surface. Forming a biofilm is a developmental process, characterized by sequential changes in gene expression and behavior as bacteria and yeast progress from discrete, free-swimming cells though stages that arrive at a mature biofilm. We are developing automated metrics to identify key transitions in early biofilm formation as cells attach to a surface, populate that surface, and adhere to each other to form early microcolonies. Our metrics use high-throughput tracking and analysis of microscopy movies to localize these transitions in space and time. Each of these transitions is associated with a loss of entropy in the bacterial system and, therefore, with biological activity that drives this loss of entropy. Better understanding of these transitions will allow automated determination of the strength and turn-on of attractive cell-surface and cell-cell interactions as biofilm development progresses.

  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. 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.

  3. Physics of amniote formation

    NASA Astrophysics Data System (ADS)

    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.

  4. Biofilm formation by enterococci.

    PubMed

    Mohamed, Jamal A; Huang, David B

    2007-12-01

    Enterococci are an important global cause of nosocomial infections, being increasingly associated with urinary tract infections, endocarditis, intra-abdominal and pelvic infections, catheter-related infections, surgical wound infections, and central nervous system infections. The two most common enterococci species are Enterococcus faecalis and Enterococcus faecium. Both are capable of producing biofilms, which consist of a population of cells attached irreversibly on various biotic and abiotic surfaces, encased in a hydrated matrix of exopolymeric substances. Many environmental and genetic factors are associated or have been proposed to be associated with the production of biofilm. This review discusses recent advances in knowledge about the biology and genetics of biofilm formation and the role of biofilms in enterococci pathogenesis.

  5. 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.

  6. 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

  7. 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

  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. Estimation of virus-neutralizing antibody in microplates.

    PubMed

    Pead, P J; Holley, A

    1973-01-01

    A simple and rapid method for the titration of virus-neutralizing antibody is described. The micro-apparatus and reagents are readily available from commercial sources. Results are included from selected cases which illustrate various conditions of immune status and response.

  10. 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.

  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. 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.

  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. 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.

  15. Formation of "bound

    NASA Astrophysics Data System (ADS)

    Nowak, K.; Kästner, M.; Miltner, A.

    2009-04-01

    During degradation of organic pollutants in soil, metabolites, microbial biomass, CO2and "bound" residues ("non-extractable" residues in soil organic matter) are formed. Enhanced transformation of these contaminants into "bound" residues has been proposed as an alternative remediation method for polluted soils. However, this kind of residues may pose a potential risk for the environment due to their chemical structure and possible remobilization under different conditions. Therefore particular attention is given actually to "bound" residues. Part of these non-extractable residues may be "biogenic," because microorganisms use the carbon from the pollutant to form their biomass components (fatty acids, amino acids, amino sugars), which subsequently may be incorporated into soil organic matter. Furthermore, the CO2 originating from mineralization of xenobiotics, can be re-assimilated by microorganisms and also incorporated into "biogenic residue". The hazard posed by "bound" residues may be overestimated because they are "biogenic" (contain microbial fatty acids and amino acids). The knowledge about the pathways of "biogenic residue" formation is necessary for a proper assessment of the fate of tested pollutants and their turnover in the soil environment. Moreover, these data are needed to establish the realistic degradation rates of the contaminants in soil. The main objectives of this study are: to quantify the extent of "biogenic residue" (fatty acids, amino acids, amino sugars) formation during the degradation of a model pollutant (2,4-dichlorophenoxyacetic acid = 2,4-D) and during CO2 assimilation by microorganisms and to evaluate which components are mainly incorporated into "bound" residues. To investigate the extent of "biogenic residue" formation in soil during the degradation of 2,4-D, experiments with either 14C-U-ring and 13C6-2,4-D or carboxyl-14C 2,4-D were performed. The incubation experiments were performed according to OECD test guideline 307, in the

  16. Prominence formation and oscillations

    NASA Astrophysics Data System (ADS)

    Chen, P. F.

    Prominences, or filaments, are a striking phenomenon in the solar atmosphere. Besides their own rich features and dynamics, they are related to many other activities, such as solar flares and coronal mass ejections (CMEs). In the past several years we have been investigating the prominence formation, oscillations, and eruptions through both data analysis and radiative hydrodynamic and magnetohydrodynamic (MHD) simulations. This paper reviews our progress on these topics, which includes: (1) With updated radiative cooling function, the coronal condensation becomes a little faster than previous work; (2) Once a seed condensation is formed, it can grow via siphon flow spontaneously even if the evaporation stops; (3) A scaling law was obtained to relate the length of the prominence thread to various parameters, indicating that higher prominences tend to have shorter threads, which is consistent with the fact that threads are long in active region prominences and short in quiescent prominences; (4) It was proposed that long-time prominence oscillations out of phase might serve as a precursor for prominence eruptions and CMEs; (5) An ensemble of oscillating prominence threads may explain the counter-streaming motion.

  17. 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

  18. Bead lightning formation

    SciTech Connect

    Ludwig, G.O.; Saba, M.M.F.

    2005-09-15

    Formation of beaded structures in triggered lightning discharges is considered in the framework of both magnetohydrodynamic (MHD) and hydrodynamic instabilities. It is shown that the space periodicity of the structures can be explained in terms of the kink and sausage type instabilities in a cylindrical discharge with anomalous viscosity. In particular, the fast growth rate of the hydrodynamic Rayleigh-Taylor instability, which is driven by the backflow of air into the channel of the decaying return stroke, dominates the initial evolution of perturbations during the decay of the return current. This instability is responsible for a significant enhancement of the anomalous viscosity above the classical level. Eventually, the damping introduced at the current channel edge by the high level of anomalous viscous stresses defines the final length scale of bead lightning. Later, during the continuing current stage of the lightning flash, the MHD pinch instability persists, although with a much smaller growth rate that can be enhanced in a M-component event. The combined effect of these instabilities may explain various aspects of bead lightning.

  19. 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".

  20. 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.

  1. 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.

  2. The KEA image file format

    NASA Astrophysics Data System (ADS)

    Bunting, Peter; Gillingham, Sam

    2013-08-01

    There are a large number of image formats already in use within the remote sensing community but currently there is no format that provides the features of: compression, support for large file sizes, ground control points, raster attribute tables and inbuilt image pyramids. Therefore, a new image format, named KEA, after the New Zealand bird, has been proposed. The KEA format provides a full implementation of the GDAL data model and is implemented within a HDF5 file. A software library with a GDAL driver have been freely provided to the community allowing use through any GDAL based software. The new format has comparable performance with existing formats while producing smaller file sizes and is already within active use for a number of projects within Landcare Research, New Zealand, and the wider community.

  3. 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

  4. Antimicrobial assay optimization and validation for HTS in 384-well format using a bioluminescent E. coli K-12 strain.

    PubMed

    Nybond, Susanna; Karp, Matti; Tammela, Päivi

    2013-07-16

    This report describes the optimization and validation of an antimicrobial assay based on the genetically modified bacterial strain Escherichia coli K-12 (pTetlux1). The use of this particular strain enables an inducible cell-based bioluminescent assay for high-throughput screening (HTS) of antimicrobial agents, which shows a pronounced detection of compounds targeting transcriptional and translational events in protein synthesis. The optimizations in 96-well format led to several improvements in assay conditions, such as reduction of the pre-incubation time before luminescence induction by half. The threshold for DMSO tolerability was concluded to be up to 1%. Assay protocol was further miniaturized into 384-well format and the liquid handling was automated using a robotic workstation. The use of compound pre-plating into 384-well plates as a part of the process was evaluated, and the total assay volume was further downscaled from 50 μl to 30 μl. With this approach, the amount of test compound needed per well was reduced to nanoliter volumes. Using the miniaturized protocol a pilot screen of 2000 known drugs and bioactives was performed. The assay performance was evaluated by calculating known assay quality parameters, the Z' factor having a mean value of 0.8 during the compound library screening indicated an excellent performance. Of the assay positives, 54 compounds showed high inhibitions (60-100%), of which the majority (89%) were known antibacterial agents. Of the actives showing >60% inhibition, 16 compounds were identified as known transcriptional and translational inhibitors. The screening results demonstrated that the miniaturized assay is well suited for identification of antimicrobial compounds in HT screening, and that the assay is specifically sensitive towards bacterial transcription and translation inhibitors.

  5. 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

  6. BACTERIOPHAGE FORMATION WITHOUT BACTERIAL GROWTH

    PubMed Central

    Price, Winston H.

    1947-01-01

    1. Iodoacetate, fluoride, and azide have been found to prevent the formation of phage and to inhibit the synthesis of ATP by Staphylococcus muscae. It is suggested that energy-rich phosphate is needed for the synthesis of phage. 2. Gramicidin prevented the formation of phage. 3. No differences were found between normal bacteria and phage-infected bacteria in the inorganic phosphate, adenosinetriphosphate, ribonucleic acid, and desoxyribonucleic acid content of the cells. 4. The mechanism of phage formation is discussed. PMID:18896936

  7. Binary stars - Formation by fragmentation

    NASA Technical Reports Server (NTRS)

    Boss, Alan P.

    1988-01-01

    Theories of binary star formation by capture, separate nuclei, fission and fragmentation are compared, assessing the success of theoretical attempts to explain the observed properties of main-sequence binary stars. The theory of formation by fragmentation is examined, discussing the prospects for checking the theory against observations of binary premain-sequence stars. It is concluded that formation by fragmentation is successful at explaining many of the key properties of main-sequence binary stars.

  8. 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.

  9. ANTIBODY FORMATION IN VITRO

    PubMed Central

    Fishman, M.

    1961-01-01

    Neutralizing activity against T2 bacteriophage appeared in cultures of lymph node cells from normal rats in response to their in vitro stimulation with a cell-free filtrate derived from homogenized rat macrophages which had been incubated with T2 bacteriophage. This activity was specifically directed against T2 bacteriophage. It resided in a fraction of the culture fluid which had the salting-out properties of serum globulin. Phage neutralization was inhibited by antibody specific for rat serum gamma globulin. Antibody production against T2 bacteriophage in cultures of lymph node cells from normal animals failed to occur if (a) T2 bacteriophage alone was added, (b) if the incubation period of macrophages and T2 phage was unduly shortened, (c) if the cell-free filtrate was heated at 80–100°C for 15 minutes, (d) if more than an optimal amount of T2 bacteriophage was added to the macrophages. Additional factors which prevented the formation of antibody were the heat inactivation of the lymph node cells or the addition to the culture medium of either streptomycin or ribonuclease. Finally, it was found that macrophages and lymph node cells had to be obtained from animals of one and the same species. All essential findings on the production of antibody to T2 bacteriophage in vitro could be confirmed by substitution of the chick embryo for the tissue culture medium. The results are discussed in terms of a possible mechanism of antibody production in which an RNAse-sensitive substance resulting from the interaction of macrophages and antigen is capable of stimulating antibody synthesis in lymphocytic cells. PMID:13893304

  10. 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.

  11. 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

  12. 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…

  13. 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?

  14. 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

  15. 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 ...

  16. 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...

  17. 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.

  18. Bibliography Formatting Software: An Update.

    ERIC Educational Resources Information Center

    Stigleman, Sue

    1993-01-01

    Discusses software used for formatting bibliographies and describes 52 programs. Information provided includes type of operating system required, publication formats, citation styles, ability to arrange by subject, use of downloaded references, network versions, costs, and plans for future changes. A directory of the programs and producers is…

  19. 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.

  20. 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.

  1. 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.

  2. Tariffs Formation on oil transportation

    NASA Astrophysics Data System (ADS)

    Glyzina, T. S.; Kolbysheva, Yu. V.; Grivtsova, I. S.; Dmitrieva, N. V.

    2016-09-01

    Oil transportation via trunk pipelines is an important part of the oil industry's activity. The main instrument of tariff regulation is the method of tariffs formation. Three methods of tariffs formation such as the method of economically justified costs (the Cost plus method), the method of economically justified return on investment capital (the RAB method), and the method of tariffs indexation were considered.

  3. 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…

  4. Sibling similarity in family formation.

    PubMed

    Raab, Marcel; Fasang, Anette Eva; Karhula, Aleksi; Erola, Jani

    2014-12-01

    Sibling studies have been widely used to analyze the impact of family background on socioeconomic and, to a lesser extent, demographic outcomes. We contribute to this literature with a novel research design that combines sibling comparisons and sequence analysis to analyze longitudinal family-formation trajectories of siblings and unrelated persons. This allows us to scrutinize in a more rigorous way whether sibling similarity exists in family-formation trajectories and whether siblings' shared background characteristics, such as parental education and early childhood family structure, can account for similarity in family formation. We use Finnish register data from 1987 through 2007 to construct longitudinal family-formation trajectories in young adulthood for siblings and unrelated dyads (N = 14,257 dyads). Findings show that family formation is moderately but significantly more similar for siblings than for unrelated dyads, also after controlling for crucial parental background characteristics. Shared parental background characteristics add surprisingly little to account for sibling similarity in family formation. Instead, gender and the respondents' own education are more decisive forces in the stratification of family formation. Yet, family internal dynamics seem to reinforce this stratification such that siblings have a higher probability to experience similar family-formation patterns. In particular, patterns that correspond with economic disadvantage are concentrated within families. This is in line with a growing body of research highlighting the importance of family structure in the reproduction of social inequality.

  5. Cyanide Formation by Chromobacterium violaceum

    PubMed Central

    Michaels, Ruth; Corpe, W. A.

    1965-01-01

    Michaels, Ruth (Columbia University, New York, N.Y.), and W. A. Corpe. Cyanide formation by Chromobacterium violaceum. J. Bacteriol. 89:106–112. 1965.—The formation of cyanide by a Chromobacterium violaceum strain was studied with growing cultures and with nonproliferating cells grown in complex and chemically defined media. Most of the cyanide was produced during the log-phase growth of the organism, and accumulated in the culture supernatant fluid. A synergistic effect of glycine and methionine on cyanide formation in a chemically defined medium was observed, and the amount of cyanide formed was found to be dependent on the concentrations of the two substances. Cyanide formation by nonproliferating cells was stimulated by preincubation with glycine and methionine. Cyanide formation by adapted cells in the presence of glycine and methionine was stimulated by succinate, malate, or fumarate, and depressed by azide and 2,4-dinitrophenol. Methionine could be replaced by betaine, dimethylglycine, and choline. PMID:14255648

  6. 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.

  7. 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

  8. 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.

  9. 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.

  10. Formate dehydrogenase from Pseudomonas oxalaticus.

    PubMed

    Müller, U; Willnow, P; Ruschig, U; Höpner, T

    1978-02-01

    Formate dehydrogenase (EC 1.2.1.2) from Pseudomonas oxalaticus has been isolated and characterized. The enzyme (molecular weight 315000) is a complex flavoprotein containing 2 FMN, 18--25 non-heme iron atoms and 15--20 acid-labile sulphides. In the last step of the purification, a sucrose gradient centrifugation, a second catalytically active species has been found apparently originating from a dissociation of the enzyme into two equal subunits. The enzyme is specific toward its natural substrate formate. It transfers electrons to NAD+, oxygen, ferricyanide, and a lot of nonphysiological acceptors (dyes). In addition electrons are transferred from NADH to these acceptors. The (reversible) removal of FMN requires a reduction step. Reincorporation has been followed by the reappearance of the reactivity against formate and by fluorescence titration. The deflavo enzyme also binds FAD and riboflavin. The resulting enzyme species show characteristic catalytic abilities. Activity against formate is peculiar to the FMN species. PMID:631130

  11. Results of Aluminosilicate Formation Testing

    SciTech Connect

    Wilmarth, W.R.

    2001-09-11

    The purpose of this work was to examine several experimental parameters of the formation of aluminosilicates under several tank chemistries, examine the conversion of crystalline phases, and determine inherent solubilities of certain crystal phases.

  12. Ultraviolet Mars Reveals Cloud Formation

    NASA Video Gallery

    Images from MAVEN's Imaging UltraViolet Spectrograph were used to make this movie of rapid cloud formation on Mars on July 9-10, 2016. The ultraviolet colors of the planet have been rendered in fal...

  13. Star Formation Across Galactic Environments

    NASA Astrophysics Data System (ADS)

    Young, Jason

    2013-01-01

    I present here parallel investigations of star formation in AGN-free and quasar host galaxies. These environments are both insightful; quasars are among the most violent objects known, reshaping their host galaxies, while my sample of AGN-free star-forming galaxies ranges from systems larger than the Milky Way to dwarf star-forming galaxies. The AGN-free galaxies are drawn from the KPNO International Spectroscopic Survey, an Hα-selected, volume-limited survey was designed to avoid continuum luminosity bias. This work studies the KISS galaxies in mid- and far-IR using Spitzer IRAC and MIPS photometry. These IR bands are interesting because the UV light from young stars is reprocessed into thermal emission in the far-IR (24μm MIPS) by dust and into vibrational transition features in the mid-IR (8.0μm IRAC) by polycyclic aromatic hydrocarbons (PAHs). This work examines the efficiencies of PAH and dust emission as tracers of star-formation. I find that the efficiency of PAH as a star-formation tracer varies with galactic stellar mass, while thermal dust has no systematic dependance on galactic mass. My study of quasar host galaxies utilizes images of eight PG quasars from the WFPC2 and NICMOS instruments aboard HST. I use narrow-band images centered on the Hβ, [OII]λ3727, [OIII]λ5007, and Paα emission lines to construct extinction and star formation maps. Additionally, I use line-ratio maps to distinguish AGN-powered line emission from star formation powered line emission. I find star formation, albeit at rates are lower than expected, suggesting that quasar host galaxies are dynamically more advanced than suspected. Seven of the galaxies have higher mass-specific star-formation rates. Additionally, I see evidence of shocked gas, supporting the hypotheses from earlier works that AGN activity quenches star formation in host galaxies by disrupting gas reservoirs.

  14. 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.

  15. 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

  16. 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.

  17. Star formation in dwarf galaxies

    NASA Astrophysics Data System (ADS)

    Dong, Shawfeng

    In this thesis, we examine the star formation history and stellar feedback effects of dwarf galaxies under the influence of extragalactic ultraviolet radiation, as well as the evolution of residual gas within tidally-limited dwarf galaxies and globular clusters. Previous work has indicated that the background UV flux can easily ionize the gas within typical dwarf galaxies, delaying or even preventing cooling and star formation within them. Many dwarf galaxies within the Local Group are, however, observed to contain multiple generations of stars, the oldest of which formed in the early epochs of cosmic evolution, when the background UV flux was intense. In order to address this paradox, we consider the dynamical evolution of gas in dwarf galaxies using a one-dimensional, spherically symmetric, Lagrangian numerical scheme which also computes the effects of radiative transfer and photoionization. We include in the scheme a physically-motivated star formation recipe and consider the effects of feedback. This scheme allows us to follow the history of the gas and of star formation within dwarf galaxies, as influenced by both external and internal UV radiation. Our results indicate that star formation in the severe environment of dwarf galaxies is a difficult and inefficient process. In potentials with total mass less than a few 106 M⊙ , and velocity dispersion less than a few km s-1 , residual gas is efficiently photoionized by cosmic background UV radiation. For intermediate mass systems, such as the dSphs around the Galaxy, star formation can proceed within early cosmic epochs despite the intense background UV flux. Triggering processes such as merger events, collisions, and tidal disturbance can lead to density enhancements, reducing the recombination timescale, allowing gas to cool and star formation to proceed. However, the star formation and gas retention efficiency may vary widely in galaxies with similar dark matter potentials, because they depend on many

  18. 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.

  19. Isolated star formation: from cloud formation to core collapse.

    PubMed

    Ward-Thompson, Derek

    2002-01-01

    The formation of stars is one of the most fundamental problems in astrophysics, as it underlies many other questions, on scales from the formation of galaxies to the formation of the solar system. The physical processes involve the turbulent behavior of a partially ionized medium containing a non-uniform magnetic field. Current debate centers around the time taken for turbulence to decay and the relative importance of the roles played by magnetic fields and turbulence. Technological advances such as millimeter-wave cameras have made possible observations of the temperature and density profiles, and statistical calculations of the lifetimes, of objects collapsing under their own self-gravity and those on the verge of collapse. Increased computing power allows more complex models to be made that include magnetic and turbulent effects. No current model can reproduce all of the observations. PMID:11778038

  20. An Adaptable Seismic Data Format

    NASA Astrophysics Data System (ADS)

    Krischer, Lion; Smith, James; Lei, Wenjie; Lefebvre, Matthieu; Ruan, Youyi; de Andrade, Elliott Sales; Podhorszki, Norbert; Bozdağ, Ebru; Tromp, Jeroen

    2016-11-01

    We present ASDF, the Adaptable Seismic Data Format, a modern and practical data format for all branches of seismology and beyond. The growing volume of freely available data coupled with ever expanding computational power opens avenues to tackle larger and more complex problems. Current bottlenecks include inefficient resource usage and insufficient data organization. Properly scaling a problem requires the resolution of both these challenges, and existing data formats are no longer up to the task. ASDF stores any number of synthetic, processed or unaltered waveforms in a single file. A key improvement compared to existing formats is the inclusion of comprehensive meta information, such as event or station information, in the same file. Additionally, it is also usable for any non-waveform data, for example, cross-correlations, adjoint sources or receiver functions. Last but not least, full provenance information can be stored alongside each item of data, thereby enhancing reproducibility and accountability. Any data set in our proposed format is self-describing and can be readily exchanged with others, facilitating collaboration. The utilization of the HDF5 container format grants efficient and parallel I/O operations, integrated compression algorithms and check sums to guard against data corruption. To not reinvent the wheel and to build upon past developments, we use existing standards like QuakeML, StationXML, W3C PROV and HDF5 wherever feasible. Usability and tool support are crucial for any new format to gain acceptance. We developed mature C/Fortran and Python based APIs coupling ASDF to the widely used SPECFEM3D_GLOBE and ObsPy toolkits.