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Sample records for 96-well plate formats

  1. 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...

  2. 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...

  3. 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.

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

    PubMed

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

    2015-11-06

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

  5. Electrothermal micromixing in 96 well plate

    NASA Astrophysics Data System (ADS)

    Kauffmann, Paul; Loire, Sophie; Mezic, Igor

    2011-11-01

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

  6. 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.

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

    PubMed

    Illges, H

    1999-01-01

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

  8. 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.

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

    PubMed

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

    2008-05-01

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

  10. High-Throughput Determination of Sodium Danshensu in Beagle Dogs by the LCMS/MS Method, Employing Liquid-Liquid Extraction Based on 96-Well Format Plates.

    PubMed

    Jiang, Jingjing; Zhao, Xin; Li, Xiuxiu; Wu, Shengyuan; Yu, Shidan; Lou, Yuefen; Fan, Guorong

    2017-04-25

    Sodium Danshensu (sodium d-(+)-β-(3,4-dihydroxyphenyl) lactate), one of the water-soluble ingredients in Salvia miltiorrhiza, exhibits potent relaxation of the coronary artery and anticoagulation effection. A high-throughput, rapid, and sensitive method combining liquid chromatography with electrospray ionization tandem mass spectrometry to determine the sodium danshensu in beagle dog plasma was developed and validated, using gallic acid as an internal standard (IS). Acidified plasma samples were extracted using 96-well liquid-liquid extraction, and were eluted on a CNW Athena C18 column (3 μm, 2.1 × 100 mm) by using a gradient mobile phase system of methanol and water (containing 0.2% formic acid). The mass spectrometric detection was achieved using negative ion electrospray ionization mode and monitoring the precursor→production combinations of m/z 197→135 for sodium danshensu and 169→125 for IS, in multiple reaction monitoring modes. Good linearity was achieved, and the linear range was 10-1000 ng/mL (R² > 0.996) with a quantification limit of 10 ng/mL for sodium danshensu in beagle dog plasma. The intra- and inter-day precision (RSD) ranged from 2.1% to 9.0%. The accuracy (RE) was between -8.6% and 5.7% at all quality control levels. The validated method was successfully applied to the pharmacokinetics study of sodium danshensu in beagle dog plasma after intravenous injection and oral administration of sodium danshensu.

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

    PubMed

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

    2004-01-27

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

  12. 96-Well plate assays for measuring collagenase activity using (3)H-acetylated collagen.

    PubMed

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

    1999-11-15

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

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

    PubMed

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

    2010-11-01

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

  14. A high-throughput sample preparation method for cellular proteomics using 96-well filter plates.

    PubMed

    Switzar, Linda; van Angeren, Jordy; Pinkse, Martijn; Kool, Jeroen; Niessen, Wilfried M A

    2013-10-01

    A high-throughput sample preparation protocol based on the use of 96-well molecular weight cutoff (MWCO) filter plates was developed for shotgun proteomics of cell lysates. All sample preparation steps, including cell lysis, buffer exchange, protein denaturation, reduction, alkylation and proteolytic digestion are performed in a 96-well plate format, making the platform extremely well suited for processing large numbers of samples and directly compatible with functional assays for cellular proteomics. In addition, the usage of a single plate for all sample preparation steps following cell lysis reduces potential samples losses and allows for automation. The MWCO filter also enables sample concentration, thereby increasing the overall sensitivity, and implementation of washing steps involving organic solvents, for example, to remove cell membranes constituents. The optimized protocol allowed for higher throughput with improved sensitivity in terms of the number of identified cellular proteins when compared to an established protocol employing gel-filtration columns. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

    PubMed

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

    2008-09-01

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

  16. 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. © 2015 Society for Laboratory Automation and Screening.

  17. Microfluidic cartridges preloaded with nanoliter plugs of reagents: an alternative to 96-well plates for screening

    PubMed Central

    Chen, Delai L; Ismagilov, Rustem F

    2006-01-01

    In traditional screening with 96-well plates, microliters of substrates are consumed for each reaction. Further miniaturization is limited by the special equipment and techniques required to dispense nanoliter volumes of fluid. Plug-based microfluidics confines reagents in nanoliter plugs (droplets surrounded by fluorinated carrier fluid), and uses simple pumps to control the flow of plugs. By using cartridges pre-loaded with nanoliter plugs of reagents, only two pumps and a merging junction are needed to set up a screen. Screening with preloaded cartridges uses only nanoliters of substrate per reaction, and requires no microfabrication. The low cost and simplicity of this method has the potential of replacing 96-well and other multi-well plates, and has been applied to enzymatic assays, protein crystallization and optimization of organic reactions. PMID:16677848

  18. 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

  19. 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…

  20. 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…

  1. Fluorescence-based liver microsomal assay for screening of pharmaceutical reactive metabolites using a glutathione conjugated 96-well plate.

    PubMed

    Ma, Xiang; Chan, Eric Chun Yong

    2010-01-01

    The purpose of our paper is to develop and validate a fluorescence-based mouse liver microsomal (MLM) assay in screening pharmaceutical reactive metabolites (RMs) using a glutathione (GSH)-conjugated 96-well plate. Poly(2-hydroxyethylmethacrylate) (pHEMA) polymeric membrane was coated on 96-well plates to provide a functional support for GSH conjugation. Oxidized GSH (GSSG) was conjugated on a cyanogen bromide (CNBr)-activated pHEMA surface. The conjugated GSH was regenerated after the reduction of GSSG using d,l-dithiothreitol (DTT). X-ray photoelectron spectroscopy, Ellman's, and fluorescence assays were applied to validate the chemistry and optimize the processes of GSH conjugation. The performance of the 96-well assay was further cross-validated using N-acetyl-p-benzo-quinone imine (NAPQI), a RM of acetaminophen (APAP), and the in vitro MLM assay of APAP. Finally, the developed method was applied to screen a batch of marketed drugs and chemicals on the formation of RMs. Our results indicated that optimum conditions were obtained for pHEMA loading, CNBr activation of pHEMA, and GSSG coupling and reduction. The detection limit of the assay for NAPQI was 500 nM with good specificity. In vitro MLM assay of APAP demonstrated a positive trapping index (TI) of 19.3%. The subsequent RM screening of a series of marketed drugs and chemical compounds resulted in a range of TI values (1.0-25.7%) that corroborated with their capacity in generating RMs. The differences of TI values are statistically significant between the compounds which are known to produce RMs and those that do not generate reactive intermediates. In conclusion, we successfully developed a fluorescence-based GSH-conjugated 96-well plate platform for the screening of RMs using MLM.

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

    PubMed

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

    2014-06-03

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

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

    PubMed Central

    2015-01-01

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

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

    PubMed

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

    2000-12-01

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

  5. Human tear analysis with miniaturized multiplex cytokine assay on "wall-less" 96-well plate.

    PubMed

    Le Guezennec, Xavier; Quah, Joanne; Tong, Louis; Kim, Namyong

    2015-01-01

    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. 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. 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 R(2) = 0.98. 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.

  6. 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

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

    PubMed

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

    2012-01-01

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

  8. 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

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

    PubMed

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

    2006-01-01

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

  10. 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.

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed

    Lai, Yinzhi; Kisaalita, William S

    2012-08-01

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

  13. 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.

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

    PubMed Central

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

    2015-01-01

    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

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

    PubMed

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

    2016-05-01

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

  17. A method for high throughput determination of viable bacteria cell counts in 96-well plates

    PubMed Central

    2012-01-01

    Background There are several methods for quantitating bacterial cells, each with advantages and disadvantages. The most common method is bacterial plating, which has the advantage of allowing live cell assessment through colony forming unit (CFU) counts but is not well suited for high throughput screening (HTS). On the other hand, spectrophotometry is adaptable to HTS applications but does not differentiate between dead and living bacteria and has low sensitivity. Results Here, we report a bacterial cell counting method termed Start Growth Time (SGT) that allows rapid and serial quantification of the absolute or relative number of live cells in a bacterial culture in a high throughput manner. We combined the methodology of quantitative polymerase chain reaction (qPCR) calculations with a previously described qualitative method of bacterial growth determination to develop an improved quantitative method. We show that SGT detects only live bacteria and is sensitive enough to differentiate between 40 and 400 cells/mL. SGT is based on the re-growth time required by a growing cell culture to reach a threshold, and the notion that this time is proportional to the number of cells in the initial inoculum. We show several applications of SGT, including assessment of antibiotic effects on cell viability and determination of an antibiotic tolerant subpopulation fraction within a cell population. SGT results do not differ significantly from results obtained by CFU counts. Conclusion SGT is a relatively quick, highly sensitive, reproducible and non-laborious method that can be used in HTS settings to longitudinally assess live cells in bacterial cell cultures. PMID:23148795

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

    PubMed

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

    2013-11-15

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

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed

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

    2015-05-14

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

  1. Rapid Determination of Ionization Constants (pK a) by UV Spectroscopy Using 96-Well Microtiter Plates.

    PubMed

    Martínez, Carlos H Ríos; Dardonville, Christophe

    2013-01-10

    We have developed a methodology that enables for the rapid measurement of ionization constants (pK a) of series of compounds by UV spectrophotometry. This protocol, which is straightforward to set up, takes advantage of the sensitivity of UV spectroscopy and the throughput enabled by the 96-well microplate (as opposed to the use of 1 cm quartz cuvette). The compounds, in stock solutions in DMSO, are dissolved in several aqueous buffer solutions directly in the microtiter plate, allowing the simultaneous determination of the UV spectra as a function of pH. Further treatment of the data provides the pK a values in a medium-throughput manner. The pK a values of 11 new antitrypanosomal dibasic compounds were determined using this methodology.

  2. Rapid depletion of dissolved oxygen in 96-well microtiter plate Staphylococcus epidermidis biofilm assays promotes biofilm development and is influenced by inoculum cell concentration.

    PubMed

    Cotter, John J; O'Gara, James P; Casey, Eoin

    2009-08-01

    Biofilm-related research using 96-well microtiter plates involves static incubation of plates indiscriminate of environmental conditions, making oxygen availability an important variable which has not been considered to date. By directly measuring dissolved oxygen concentration over time we report here that dissolved oxygen is rapidly consumed in Staphylococcus epidermidis biofilm cultures grown in 96-well plates irrespective of the oxygen concentration in the gaseous environment in which the plates are incubated. These data indicate that depletion of dissolved oxygen during growth of bacterial biofilm cultures in 96-well plates may significantly influence biofilm production. Furthermore higher inoculum cell concentrations are associated with more rapid consumption of dissolved oxygen and higher levels of S. epidermidis biofilm production. Our data reveal that oxygen depletion during bacterial growth in 96-well plates may significantly influence biofilm production and should be considered in the interpretation of experimental data using this biofilm model.

  3. A rubber transfer gasket to improve the throughput of liquid-liquid extraction in 96-well plates: application to vitamin D testing.

    PubMed

    Hoofnagle, Andrew N; Laha, Thomas J; Donaldson, Thomas F

    2010-06-01

    The unmitigated rise in demand for the assessment of vitamin D status has taxed the ability of clinical mass spectrometry laboratories to preserve turn-around times. We aimed to improve the throughput of liquid-liquid extraction of plasma/serum for the assay of 25-hydroxy vitamin D. We designed and fabricated a flexible rubber gasket that seals two 96-well plates together to quantitatively transfer the contents of one plate to another. Using the transfer gasket and a dry-ice acetone bath to freeze the aqueous infranatant, we developed a novel liquid-liquid extraction workflow in a 96-well plate format. We applied the technology to the mass spectrometric quantification of 25-hydroxy vitamin D. Cross-contamination between wells was < or = 0.13%. The interassay imprecision over 132 days of clinical implementation was less than 10%. The method compared favorably to a standard liquid-liquid extraction in glass tubes (Deming slope=1.018, S(x|y)=0.022). The accuracy of the assay was 102-105% as assessed with the recently released control materials from NIST. The development of a plate-sealing gasket permits the liquid-liquid extraction of clinical specimens in a moderate-throughput workflow and the reliable assay of vitamin D status. In the future, the gasket may also prove useful in other sample preparation techniques for HPLC or mass spectrometry. Copyright 2010 Elsevier B.V. All rights reserved.

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

    PubMed

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

    2016-11-25

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

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

    PubMed Central

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

    2016-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-11-01

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

  7. 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

    2015-06-07

    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

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

    PubMed

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

    2011-02-28

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

  9. High-Throughput Lipolysis in 96-Well Plates for Rapid Screening of Lipid-Based Drug Delivery Systems.

    PubMed

    Mosgaard, Mette D; Sassene, Philip J; Mu, Huiling; Rades, Thomas; Müllertz, Anette

    2017-04-01

    The high-throughput in vitro intestinal lipolysis model (HTP) applicable for rapid and low-scale screening of lipid-based drug delivery systems (LbDDSs) was optimized and adjusted as to be conducted in 96-well plates (HTP-96). Three different LbDDSs (I-III) loaded with danazol or cinnarizine were used as model systems. The distributions of cinnarizine and danazol in the aqueous and precipitated digestion phases generated during lipolysis in HTP-96 were compared with previously published data obtained from HTP. The final HTP-96 setup resulted in the same rank order as the original HTP model with regard to solubilization in the aqueous phase during digestion: LbDDS III > LbDDS II > LbDDS I for danazol and LbDDS III ≈ LbDDS II ≈ LbDDS I for cinnarizine. HTP-96 is a useful model for fast performance assessment of LbDDS in a small scale. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  10. 96-well plate-to-plate gravity fluorous solid-phase extraction (F-SPE) for solution-phase library purification.

    PubMed

    Zhang, Wei; Lu, Yimin

    2007-01-01

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

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

    PubMed

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

    2009-04-01

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

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

    PubMed Central

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

    2017-01-01

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

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

    PubMed Central

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

    2010-01-01

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

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

    PubMed

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

    2000-03-01

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

  15. 96-Well Polycarbonate-Based Microfluidic Titer Plate for High-Throughput Purification of DNA and RNA

    PubMed Central

    Witek, Małgorzata A.; Hupert, Mateusz L.; Park, Daniel S.-W.; Fears, Kirby; Murphy, Michael C.; Soper, Steven A.

    2008-01-01

    We report a simple and effective method for the high-throughput purification of a variety of nucleic acids (NAs) from whole cell lysates or whole blood using a photactivated polycarbonate solid-phase reversible immobilization (PPC-SPRI) microfluidic chip. High-throughput operation was achieved by placing 96 purification beds, each containing an array of 3800 20 µm diameter posts, on a single 3″ × 5″ polycarbonate (PC) wafer fabricated by hot embossing. All beds were interconnected through a common microfluidic network that permitted parallel access through the use of a vacuum and syringe pump for delivery of immobilization buffer (IB) and effluent. The PPC-SPRI purification was accomplished by condensation of NAs onto a UV-modified PC surface in the presence of the IB comprised of polyethylene glycol, NaCl, and ethanol with a composition dependent on the length of the NAs to be isolated and the identity of the sample matrix. The performance of the device was validated by quantification of the recovered material following PCR (for DNA) or RT-PCR (for RNA). The extraction bed load capacity of NAs was 206 ± 93 ng for gDNA and 165 ± 81 ng for TRNA from Escherichia coli. Plate-to-plate variability was found to be 35 ± 10%. The purification process was fast (>30 min) and easy to automate, and the low cost of wafer fabrication makes it appropriate for single-use applications. PMID:18355091

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

    PubMed

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

    2013-01-30

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

  17. High-throughput synthesis of conopeptides: a safety-catch linker approach enabling disulfide formation in 96-well format.

    PubMed

    Brust, Andreas; Tickle, Alice E

    2007-02-01

    Conotoxins exhibit a high degree of selectivity and potency for a range of pharmacologically relevant targets. The rapid access to libraries of conotoxin analogues, containing multiple intramolecular disulfide bridges for use in drug development, can be a very labor intensive, multi-step task. This work describes a high-throughput method for the synthesis of cystine-bridged conopeptides. Peptides were assembled on a peptide synthesizer employing the Fmoc solid-phase strategy using a safety-catch amide linker (SCAL). Side-chain protecting groups were removed on solid phase before SCAL activation with ammonium iodide in TFA, finally releasing the peptide into the TFA solution. Disulfide bond formation was performed in the cleavage mixture employing DMSO. This improved method allows mixtures of oxidized peptides to be obtained in parallel directly from a peptide synthesizer. A single HPLC purification of the resulting crude oxidized material produced peptides of > 95% purity.

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

    PubMed

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

    2012-08-31

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

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

    PubMed

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

    1999-08-27

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

  20. 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...

  1. 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...

  2. 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.

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

    PubMed

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

    2006-09-01

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

  4. Development and validation of a method for fipronil residue determination in ovine plasma using 96-well plate solid-phase extraction and gas chromatography-tandem mass spectrometry.

    PubMed

    Bichon, E; Richard, C A; Le Bizec, B

    2008-08-01

    Fipronil, a phenylpyrazole insecticide introduced for pest control on a broad range of crops, undergoes a reinforcement of the regulation within the European Union (2007/52/EC directive) due to its potential effects on environment and human health. In order to assess the plasmatic concentrations of fipronil residues (sulfone, sulfide, fipronil, desulfinyl and amide) in ovine, a methodology based on gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) was developed and validated according to the European standard (2002/657/EC). The proposed method allows a large number of samples to be treated concurrently (n=80) using a reduced sample amounts (0.2 mL), and consents to reach a level of quantification of 0.1 pg microL(-1). The sample preparation consisted of a single solid-phase extraction (SPE) purification on a 96-well plate filled with a styrene-divinyl-benzene phase. Linearity was demonstrated all along the investigated range of concentrations, i.e. from 0.25 to 2000 pg microL(-1), with coefficient of determination (R(2)) from 0.977 to 0.994, depending on target analytes. Calculated decision limit (CCalpha) and detection capability (CCbeta) for fipronil, sulfone and sulphide were in the range 0.05-0.16 and 0.28-0.73 pg microL(-1) respectively.

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

    PubMed

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

    2012-06-01

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

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

    PubMed

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

    2011-07-01

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

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

    PubMed

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

    2008-08-01

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

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

    PubMed

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

    2015-01-01

    Basidiomycete strains synthesize several types of β-d-glucans, which play a major role in the medicinal properties of mushrooms. Therefore, the specific quantification of these β-d-glucans in mushroom strains is of great biochemical importance. Because published assay methods for these β-d-glucans present some disadvantages, a novel colorimetric assay method for β-d-glucan with alcian blue dye was developed. The complex formation was detected by following the decrease in absorbance in the range of 620 nm and by hypsochromic shift from 620 to 606 nm (∼14 nm) in UV-Vis spectrophotometer. Analysis of variance was used for optimization of the slope of the calibration curve by using the assay mixture containing 0.017% (w/v) alcian blue in 2% (v/v) acetic acid at pH 3.0. The high-throughput colorimetric assay method on microtiter plates was used for quantification of β-d-glucans in the range of 0-0.8 μg, with a slope of 44.15 × 10(-2) and a limit of detection of 0.017 μg/well. Recovery experiments were carried out by using a sample of Hericium erinaceus, which exhibited a recovery of 95.8% for β-1,3-d-glucan. The present assay method exhibited a 10-fold higher sensitivity and a 59-fold lower limit of detection compared with the published method with congo red. β-d-glucans of several mushrooms strains were isolated from fruiting bodies and mycelia, and they were quantified by this assay method. This assay method is fast, specific, simple, and it can be used to quantify β-d-glucans from other biological sources. © 2015 American Institute of Chemical Engineers.

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

    PubMed

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

    2011-02-01

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

  10. A 96-well microtiter plate assay for high-throughput screening of Mycobacterium tuberculosis dTDP-d-glucose 4,6-dehydratase inhibitors.

    PubMed

    Shi, Xiaoxia; Sha, Shanshan; Liu, Likun; Li, Xin; Ma, Yufang

    2016-04-01

    Mycobacterium tuberculosis dTDP-d-glucose 4,6-dehydratase (RmlB) is the second enzyme for the biosynthesis of dTDP-l-rhamnose, which is a sugar donor to the synthesis of the cell wall linker, d-N-acetylglucosamine-l-rhamnose. RmlB is essential to mycobacterial growth and is not found in humans; therefore, it is a potential target for developing new anti-tuberculosis drugs. So far, there has been no suitable method for high-throughput screening of RmlB inhibitors. Here, the recombinant M. tuberculosis RmlB was purified and an absorbance-based microtiter plate assay was developed for RmlB activity. It could be used for high-throughput screening of RmlB inhibitors. The kinetic properties of M. tuberculosis RmlB, including optimal pH, optimal temperature, the effect of metal ions, and the kinetic parameters, were determined with this assay. The inhibitory effects of dTTP and dTDP on M. tuberculosis RmlB were also studied with the assay. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. 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. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. A Pitfall of the 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethonyphenol)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay due to evaporation in wells on the edge of a 96 well plate.

    PubMed

    Patel, Manish I; Tuckerman, Rodney; Dong, Qihan

    2005-06-01

    The 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethonyphenol)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) calorimetric assay is replacing the traditional 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as a fast, one-step assay of cell viability. We have observed that evaporation of the outer wells of a 96 well plate increases the absorbancy by 52% compared to the inner wells. Filling the outer 2 rows of wells with media and replacement of the media prior to addition of the MTS reagent will, however, correct this inaccuracy.

  13. 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.

  14. Two new techniques for sample preparation in bioanalysis: microextraction in packed sorbent (MEPS) and use of a bonded monolith as sorbent for sample preparation in polypropylene tips for 96-well plates.

    PubMed

    Blomberg, Lars G

    2009-02-01

    Analytical methods providing high throughput are required for the ever increasing number of samples in bioanalysis. Currently, the method of choice in bioanalysis is LC-MS-MS. This method is quite rapid and thereby the focus has been directed to sample preparation as being a bottleneck in total analysis systems. It has become necessary to develop sample preparation techniques to a new improved level. This development has been based on a systematic and scientific approach. The key factors in this development have been miniaturization, integration, and automation of the techniques. This review provides a short overview of recent developments. Special emphasis is on two techniques: microextraction in packed syringe (MEPS) and use of a monolithic acrylamide plug as sorbent in polypropylene tips primarily intended for use with 96-well plate systems.

  15. 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.

  16. A novel approach to the simultaneous extraction and non-targeted analysis of the small molecules metabolome and lipidome using 96-well solid phase extraction plates with column-switching technology.

    PubMed

    Li, Yubo; Zhang, Zhenzhu; Liu, Xinyu; Li, Aizhu; Hou, Zhiguo; Wang, Yuming; Zhang, Yanjun

    2015-08-28

    This study combines solid phase extraction (SPE) using 96-well plates with column-switching technology to construct a rapid and high-throughput method for the simultaneous extraction and non-targeted analysis of small molecules metabolome and lipidome based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry. This study first investigated the columns and analytical conditions for small molecules metabolome and lipidome, separated by an HSS T3 and BEH C18 columns, respectively. Next, the loading capacity and actuation duration of SPE were further optimized. Subsequently, SPE and column switching were used together to rapidly and comprehensively analyze the biological samples. The experimental results showed that the new analytical procedure had good precision and maintained sample stability (RSD<15%). The method was then satisfactorily applied to more widely analyze the small molecules metabolome and lipidome to test the throughput. The resulting method represents a new analytical approach for biological samples, and a highly useful tool for researches in metabolomics and lipidomics. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    PubMed

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

    2016-02-01

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

  18. Rapid and Specific Approach for Direct Measurement of Glimepiride in Human Plasma by LC–ESI-MS–MS Employing Automated 96 Well Format: Application to a Bioequivalence Study

    PubMed Central

    Kundlik, M.L.; Zaware, B.H.; Kuchekar, S.R.

    2012-01-01

    A rapid liquid chromatographic method with electrospray ionization tandem mass spectrometric (LC–MS–MS) detection is developed and validated for quantification of glimepiride in heparinized human plasma. Plasma samples, without a drying and reconstitution step, are extracted by solid-phase extraction (SPE) and eluted with 0.9 mL of acetonitrile–methanol (1:1, v/v) containing 0.05% formic acid. The analyte and glimepiride d8 (internal standard, IS) are chromatographed on a C18 column; the mobile phase is acetonitrile–2 mm ammonium formate (88:12, v/v), with the pH adjusted to 3.5 with formic acid, at a flow rate of 0.5 mL/min. The retention times of glimepiride and the IS are 0.93 min, and the runtime is 1.6 min per sample. Selected reaction monitoring of MH+ at m/z 491.20 and 499.26 result in stable fragment ions with m/z 351.80 and 359.96 for glimepiride and the IS, respectively. The response was a linear function of the concentration in the range of 2.0–650.0 ng/mL, with r ≥ 0.9994. The recovery of glimepiride and the IS ranged from 81.91 to 83.36%. The assay has excellent characteristics and has been successfully used for the analysis of glimepiride in healthy human subjects in a bioequivalence study. It was well suited to clinical studies of the drug involving large numbers of samples. PMID:22291058

  19. 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. Copyright © 2012. Published by Elsevier B.V.

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

    PubMed Central

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

    2015-01-01

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

  1. 96-well electroporation method for transfection of mammalian central neurons.

    PubMed

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

    2006-11-01

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

  2. 96-Well electroporation method for transfection of mammalian central neurons

    PubMed Central

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

    2008-01-01

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

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

    PubMed

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

    2001-02-25

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

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

    PubMed

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

    2014-05-30

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

  5. Development and validation of a rapid HPLC method for the determination of cefdinir in beagle dog plasma integrated with an automatic on-line solid-phase extraction following protein precipitation in the 96-well plate format.

    PubMed

    Li, Ji; Wang, Li; Chen, Zhao; Xie, Rui; Li, You; Hang, Taijun; Fan, Guorong

    2012-05-01

    The high-performance liquid chromatography (HPLC) coupled with on-line solid phase extraction (SPE) and ultraviolet (UV) detection was developed for determining cefdinir in beagle dog plasma. After simple pretreatment for plasma with 6% perchloric acid, a volume of 100 μL upper layer of the plasma sample was injected into the self-made on-line SPE column. The analytes were retained on the trap column (Lichrospher C(18), 4.6 mm × 37 mm, 25 μm), and the biological matrix was washed out with the solvent (20mM KH(2)PO(4) adjusted pH 3.0) at flow rate of 2 mL/min. By rotation of the switching valve, the target analytes could be eluted from trap column to analytical column in the back-flush mode by the mobile phase (methanol-acetonitrile-20mM KH(2)PO(4) adjusted pH 3.0, 11.25:6.75:82, v/v/v) at flow rate of 1.5 mL/min, and then separated on the analytical column (Ultimate XB-C(18), 4.6 mm × 50 mm, 5 μm). The complete cycle of the on-line SPE preconcentration, purification and HPLC separation of the analytes was 4 min. The UV detection was performed at 286 nm. The calibration curves showed excellent linear relationship (R(2)=0.9995) over the concentration range of 0.05-50 μg/mL. The optimized method showed good performance in terms of specificity, linearity, detection and quantification limits, precision and accuracy. This method was successfully applied to quantify cefdinir in beagle dog plasma to support the pre-clinical pharmacokinetic trial. Copyright © 2012. Published by Elsevier B.V.

  6. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  9. 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.

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

    PubMed

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

    2017-02-27

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

  11. 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.

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

    PubMed Central

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

    2017-01-01

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

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

    PubMed

    Hudlow, William R; Buoncristiani, Martin R

    2012-01-01

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

  14. LOWLID FORMATION AND PLATE TECTONICS ON EXOPLANETS

    NASA Astrophysics Data System (ADS)

    Stamenkovic, V.; Noack, L.; Breuer, D.

    2009-12-01

    The last years of astronomical observation have opened the doors to a universe filled with extrasolar planets. Detection techniques still only offer the possibility to detect mainly Super-Earths above five Earth masses. But detection techniques do steadily improve and are offering the possibility to detect even smaller planets. The observations show that planets seem to exist in many possible sizes just as the planets and moons of our own solar system do. It is only a natural question to ask if planetary mass has an influence on some key habitability factors such as on plate tectonics, allowing us to test which exoplanets might be more likely habitable than others, and allowing us to understand if plate tectonics on Earth is a stable or a critical, instable process that could easily be perturbed. Here we present results derived from 1D parameterized thermal evolution and 2D/3D computer models, showing how planetary mass influences the propensity of plate tectonics for planets with masses ranging from 0.1 to 10 Earth masses. Lately [2, 3] studied the effect of planetary mass on the ability to break plates and hence initiate plate tectonics - but both derived results contradictory to the other. We think that one of the reasons why both studies [2, 3] are not acceptable in their current form is partly due to an oversimplification. Both treated viscosity only temperature-dependent but neglected the effect pressure has on enlarging the viscosity in the deep mantle. More massive planets have therefore a stronger pressure-viscosity-coupling making convection at high pressures sluggish or even impossible. For planets larger than two Earth masses we observe that a conductive lid (termed low-lid) forms above the core-mantle boundary and thus reduces the effective convective part of the mantle when including a pressure-dependent term into the viscosity laws as shown in [1]. Moreover [2, 3] use time independent steady state models neglecting the fact that plate tectonics is a

  15. Droplet formation under the effect of a flexible nozzle plate.

    PubMed

    Sangplung, S; Liburdy, J A

    2009-09-01

    Droplet formation from a flexible nozzle plate driven by a prescribed-waveform excitation of a piezoelectric is numerically investigated using a computational fluid dynamics (CFD) model with the volume of fluid (VOF) method. The droplet generator with a flexible nozzle plate, which is free to vibrate due to the pressure acting on the plate, is modeled in a CFD computational domain. The CFD analysis includes the fluid-structure interaction between fluid and a flexible plate using large deflection theory. The problem is characterized by the nondimensional variables based on the capillary parameters of time, velocity, and pressure. The CFD model is validated with the experiment results. This study examines the characteristics of the applied waveforms and nozzle plate material properties to change the vibrational characteristics of the nozzle plate. The effect of fluid properties on the droplet formation process is also investigated focusing on surface tension and viscous forces. Increasing the impulse of the piezoelectric can be used to cause a higher droplet velocity and it is shown that the vibration of the nozzle plate has a strong effect on the droplet velocity, shape, and volume. Surface tension has a strong influence on the droplet formation characteristics in contrast to viscous forces. For the combination of a fluid with high surface tension and the most flexible nozzle plate, this system cannot cause the droplet ejected out of the nozzle.

  16. Jet formation at interaction of a vibrating plate with liquid

    NASA Astrophysics Data System (ADS)

    Aleksandrov, V. A.; Kopysov, S. P.; Tonkov, L. Y.

    2017-06-01

    In this work, we experimentally investigate the mechanism of jet formation on liquid surface near the edge of a vibrating plate that is partially immersed in the liquid. Under the conditions of resonant bending vibrations, the vibrating plate excites capillary waves in the form of Faraday ripples on the surface of the liquid layer wetting the plate surface. Two-dimensional capillary waves also appear on the curved surface of the liquid near the edges of the vibrating plate. The vibrations of the plate area generate hydrodynamic pressure on the liquid surface, which initiates surface eddy flows. At a certain position of the vibrating plate in the liquid, capillary oscillations in the form of standing waves appear along the boundary of the wetting layer of the liquid, directly under the free edge of the plate. The vibrations of the plate edge modulate the standing waves in the transverse direction, which results in the periodic variation of the surface curvature of the wetting liquid layer, from negative to positive values. The inertia forces, periodically varying with the frequency of the plate vibrations, combined with the Laplace pressure, in the excited standing capillary waves on the surface flow under the plate edge, initiates the periodic ejection of particles of the liquid forming a jet.

  17. Formation of vortex wakes at flow separation from plate

    NASA Astrophysics Data System (ADS)

    Gorelov, D. N.; Govorova, A. I.

    2017-05-01

    The plane nonlinear initial boundary value problem about the separated flow past a plate set in motion at a constant velocity from the state of rest has been considered. Results of a numerical experiment which have allowed us to trace in detail the vortex-wake formation process behind a vertical plate are reported. It is shown that, after the beginning of the plate motion, several stable vortical structures, including a Karman street, form in succession behind the plate. It is found that, on the emergence of the Karman street, there occurs a sharp and substantial growth of vortex-wake intensity and hydrodynamic drag force with a pulsating time behavior. A conclusion about the origination, in this regime, of self-sustained oscillations of the liquid in the vicinity of the plate is drawn.

  18. A universal plate format for increased throughput of assays that monitor multiple aminoacyl-tRNA synthetase activities

    PubMed Central

    Beebe, Kirk; Waas, William; Druzina, Zhanna; Guo, Min; Schimmel, Paul

    2013-01-01

    Aminoacyl tRNA synthetases are intensely studied enzymes because of their importance in the establishment of the genetic code and their connection to disease and medicine. During the advancement of this field, several assays were developed. Despite many innovations, the sensitivity, simplicity, and reliability of the radiometric assays (that were among the first to be developed) have ensured their continued use. Four activities are measured by these assays: active site titration, amino acid activation, aminoacylation, and post-transfer editing (deacylation). In an effort to maintain the advantage of these assays, while enhancing throughput, reducing waste, and improving data quality, a universal 96-well filter plate format was developed. This format facilitates the assays for all 4 of the widely studied activities. PMID:17603003

  19. A universal plate format for increased throughput of assays that monitor multiple aminoacyl transfer RNA synthetase activities.

    PubMed

    Beebe, Kirk; Waas, William; Druzina, Zhanna; Guo, Min; Schimmel, Paul

    2007-09-01

    Aminoacyl transfer RNA (tRNA) synthetases are intensely studied enzymes because of their importance in the establishment of the genetic code and their connection to disease and medicine. During the advancement of this field, several assays were developed. Despite many innovations, the sensitivity, simplicity, and reliability of the radiometric assays (which were among the first to be developed) have ensured their continued use. Four activities are measured by these assays: active site titration, amino acid activation, aminoacylation, and posttransfer editing (deacylation). In an effort to maintain the advantage of these assays while enhancing throughput, reducing waste, and improving data quality, a universal 96-well filter plate format was developed. This format facilitates the assays for all four of the widely studied activities.

  20. Signaling pathways regulating cartilage growth plate formation and activity.

    PubMed

    Samsa, William E; Zhou, Xin; Zhou, Guang

    2017-02-01

    The growth plate is a highly specialized and dynamic cartilage structure that serves many essential functions in skeleton patterning, growth and endochondral ossification in developing vertebrates. Major signaling pathways initiated by classical morphogens and by other systemic and tissue-specific factors are intimately involved in key aspects of growth plate development. As a corollary of these essential functions, disturbances in these pathways due to mutations or environmental factors lead to severe skeleton disorders. Here, we review these pathways and the most recent progress made in understanding their roles in chondrocyte differentiation in growth plate development and activity. Furthermore, we discuss newly uncovered pathways involved in growth plate formation, including mTOR, the circadian clock, and the COP9 signalosome.

  1. Multiwell cell culture plate format with integrated microfluidic perfusion system

    NASA Astrophysics Data System (ADS)

    Domansky, Karel; Inman, Walker; Serdy, Jim; Griffith, Linda G.

    2006-01-01

    A new cell culture analog has been developed. It is based on the standard multiwell cell culture plate format but it provides perfused three-dimensional cell culture capability. The new capability is achieved by integrating microfluidic valves and pumps into the plate. The system provides a means to conduct high throughput assays for target validation and predictive toxicology in the drug discovery and development process. It can be also used for evaluation of long-term exposure to drugs or environmental agents or as a model to study viral hepatitis, cancer metastasis, and other diseases and pathological conditions.

  2. Eastern Indian Ocean microcontinent formation driven by plate motion changes

    NASA Astrophysics Data System (ADS)

    Whittaker, J. M.; Williams, S. E.; Halpin, J. A.; Wild, T. J.; Stilwell, J. D.; Jourdan, F.; Daczko, N. R.

    2016-11-01

    The roles of plate tectonic or mantle dynamic forces in rupturing continental lithosphere remain controversial. Particularly enigmatic is the rifting of microcontinents from mature continental rifted margins, with plume-driven thermal weakening commonly inferred to facilitate calving. However, a role for plate tectonic reorganisations has also been suggested. Here, we show that a combination of plate tectonic reorganisation and plume-driven thermal weakening were required to calve the Batavia and Gulden Draak microcontinents in the Cretaceous Indian Ocean. We reconstruct the evolution of these two microcontinents using constraints from new paleontological samples, 40Ar/39Ar ages, and geophysical data. Calving from India occurred at 101-104 Ma, coinciding with the onset of a dramatic change in Indian plate motion. Critically, Kerguelen plume volcanism does not appear to have directly triggered calving. Rather, it is likely that plume-related thermal weakening of the Indian passive margin preconditioned it for microcontinent formation but calving was triggered by changes in plate tectonic boundary forces.

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

    PubMed

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

    2004-02-04

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

  4. Development and Testing of a Colorimetric 96 Well Plate Assay for the Determination of HD Hydrolysis Rate in Various Formulations

    DTIC Science & Technology

    2005-01-01

    Cold Foam 1% ethylene glycol 1% benzalkonium chloride 1% polyvinylpolypyrrolidone 1% Brij 58 1% Benzyldimethyl tetradecyl ammonium chloride dihydrate...LITERA TURE CITED ..................................................................................... 15 5 FIGURES 1. Rates of Chloride Release in...Ground, MD). The HD was stabilized with tributylamine and was approximately 90% pure. Quantitative assays were conducted with a chloride electrode

  5. High Speed Large Format Photon Counting Microchannel Plate Imaging Sensors

    NASA Astrophysics Data System (ADS)

    Siegmund, O.; Ertley, C.; Vallerga, J.; Craven, C.; Popecki, M.; O'Mahony, A.; Minot, M.

    The development of a new class of microchannel plate technology, using atomic layer deposition (ALD) techniques applied to a borosilicate microcapillary array is enabling the implementation of larger, more stable detectors for Astronomy and remote sensing. Sealed tubes with MCPs with SuperGenII, bialkali, GaAs and GaN photocathodes have been developed to cover a wide range of optical/UV sensing applications. Formats of 18mm and 25mm circular, and 50mm (Planacon) and 20cm square have been constructed for uses from night time remote reconnaissance and biological single-molecule fluorescence lifetime imaging microscopy, to large area focal plane imagers for Astronomy, neutron detection and ring imaging Cherenkov detection. The large focal plane areas were previously unattainable, but the new developments in construction of ALD microchannel plates allow implementation of formats of 20cm or more. Continuing developments in ALD microchannel plates offer improved overall sealed tube lifetime and gain stability, and furthermore show reduced levels of radiation induced background. High time resolution astronomical and remote sensing applications can be addressed with microchannel plate based imaging, photon time tagging detector sealed tube schemes. Photon counting imaging readouts for these devices vary from cross strip (XS), cross delay line (XDL), to stripline anodes, and pad arrays depending on the intended application. The XS and XDL readouts have been implemented in formats from 22mm, and 50mm to 20cm. Both use MCP charge signals detected on two orthogonal layers of conductive fingers to encode event X-Y positions. XDL readout uses signal propagation delay to encode positions while XS readout uses charge cloud centroiding. Spatial resolution readout of XS detectors can be better than 20 microns FWHM, with good image linearity while using low gain (<10^6), allowing high local counting rates and longer overall tube lifetime. XS tubes with electronics can encode event

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

    PubMed

    Yang, Minghui; Sun, Steven; Kostov, Yordan; Rasooly, Avraham

    2011-03-31

    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.

  7. Vortex Formation Behind an Inclined 2-Dimensional Thin Flat Plate

    NASA Astrophysics Data System (ADS)

    Mohebi, Meraj; Wood, David H.; Martinuzzi, Robert J.

    2014-11-01

    Stereo Particle Image Velocimetry was used to measure the turbulent wake of a 2D flat plate inclined relative to a uniform stream as a heuristic model for airfoils and wind turbine blades at high incidence. Phase Averaging was performed to study the vortex dynamics and relate these to the force characteristics. Below 90°, immediately behind the plate, rounder and more organized trailing edge vortices form which possess higher circulation and are associated with higher Reynolds stresses than the counter-rotating, weaker and elongated leading edge vortices. The quasi-periodically shed vortices on the sides of the wake decay in strength at different rates to reach a circulation ratio of -1 within a distance less than 5 chords downstream of the plate for all angles. This equalization of vortex strength is related to an increase in turbulence diffusion, due to mostly-incoherent 3-dimensionality which progressively increases as the inclination angle is reduced, and convective transfer of vorticity between counter-rotating vortices. The wake experiences a sudden change in vortex formation mechanism at around 40°. At this angle, the frequency analysis on the signals of a pair of micro-pressure transducers in the wake also shows a discontinuity in the trends. This work was supported by NSERC Discovery grants to R. J. Martinuzzi and D. H. Wood.

  8. 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.

  9. Formation of plate boundaries: The role of mantle volatilization

    NASA Astrophysics Data System (ADS)

    Seno, Tetsuzo; Kirby, Stephen H.

    2014-02-01

    In the early Earth, convection occurred with the accumulation of thick crust over a weak boundary layer downwelling into the mantle (Davies, G.F., 1992. On the emergence of plate tectonics. Geology 20, 963-966.). This would have transitioned to stagnant-lid convection as the mantle cooled (Solomatov, V.S., Moresi, L.-N., 1997. Three regimes of mantle convection with non-Newtonian viscosity and stagnant lid convection on the terrestrial planets. Geophys. Res. Lett. 24, 1907-1910.) or back to a magma ocean as the mantle heated (Sleep, N., 2000. Evolution of the mode of convection within terrestrial planets. J. Geophys. Res. 105(E7): 17563-17578). Because plate tectonics began operating on the Earth, subduction must have been initiated, thus avoiding these shifts. Based on an analogy with the continental crust subducted beneath Hindu Kush and Burma, we propose that the lithosphere was hydrated and/or carbonated by H2O-CO2 vapors released from magmas generated in upwelling plumes and subsequently volatilized during underthrusting, resulting in lubrication of the thrust above, and subduction of the lithosphere along with the overlying thick crust. Once subduction had been initiated, serpentinized forearc mantle may have formed in a wedge-shaped body above a dehydrating slab. In relict arcs, suture zones, or rifted margins, any agent that warms and dehydrates the wedge would weaken the region surrounding it, and form various types of plate boundaries depending on the operating tectonic stress. Thus, once subduction is initiated, formation of plate boundaries might be facilitated by a major fundamental process: weakening due to the release of pressurized water from the warming serpentinized forearc mantle.

  10. Metamorphic sole formation reveals plate interface rheology during early subduction

    NASA Astrophysics Data System (ADS)

    Mathieu, S.; Agard, P.; Dubacq, B.; Plunder, A.; Prigent, C.

    2015-12-01

    Metamorphic soles are m to ~500m thick tectonic slices welded beneath most large ophiolites. They correspond to highly to mildly deformed portions of oceanic lithosphere metamorphosed at amphibolite to granulite facies peak conditions. Metamorphic soles are interpreted as formed ≤1-2Ma after intraoceanic subduction initiation by heat transfer from the hot, incipient mantle wegde to the underthrusting lower plate. Their early accretion and exhumation together with the future ophiolite implies at least one jump of the subduction plate interface from above to below the metamorphic sole. Metamorphic soles thus represent one of the few remnants of the very early evolution of the subduction plate interface and provide major constraints on the thermal structure and the effective rheology of the crust and mantle along the nascent slab interface.We herein present a structural and petrological detailed description of the Oman and Turkey metamorphic soles. Both soles present a steep inverted metamorphic structure, with isograds subparallel to the peridotite contact, in which the proportion of mafic rocks, pressure and temperature conditions increase upward. They comprise, as most metamorphic soles worldwide, two main units: (1) a high-grade unit adjacent to the overlying peridotite composed of granulitized to amphibolized metabasalts, with rare metasedimentary interlayers (~800±100ºC at 10±2kbar) and (2) a low-grade greenschist facies unit composed of metasedimentary rocks with rare metatuffs (~500±100ºC at 5±2kbar). We provide for the first time refined P-T peak condition estimations by means of pseudosection modelling and maximum temperature constraints for the Oman low-grade sole by RAMAN thermometry. In order to quantify micro-scale deformations trough the sole, we also present EBSD data on the Oman garnet-bearing and garnet-free high-grade sole.With these new constraints, we finally propose a new conceptual mechanical model for metamorphic sole formation. This

  11. Grain damage, phase mixing and plate-boundary formation

    NASA Astrophysics Data System (ADS)

    Bercovici, David; Skemer, Philip

    2017-07-01

    The generation of plate tectonics on Earth relies on complex mechanisms for shear localization, as well as for the retention and reactivation of weak zones in the cold ductile lithosphere. Pervasive mylonitization, wherein zones of high deformation coincide with extensive mineral grain size reduction, is an important clue to this process. In that regard, the grain-damage model of lithospheric weakening provides a physical framework for both mylonitization and plate generation, and accounts for the competition between grain size reduction by deformation and damage, and healing by grain growth. Zener pinning at the evolving interface between mineral components, such as olivine and pyroxene, plays a key role in helping drive grains to small mylonitic sizes during deformation, and then retards their growth once deformation ceases. The combined effects of damage and pinning, however, rely on the efficiency of inter-grain mixing between phases (e.g., olivine and pyroxene) and grain dispersal, which likely depends on grain size itself. Here we present a new model for inter-grain mixing and damage and the onset of rapid mixing. The model considers the competition between the formation of new grains behind a receding interphase triple junction (e.g., olivine growing into a boundary between two pyroxene grains) and their severance or spalling during progressive deformation and damage. The newly formed grains of one phase are then transported along the opposing phase's grain-boundaries and the two phases become dispersed at the grain-scale in a growing mixed layer. The small intermixed grains also affect the grain evolution of the surrounding host grains by Zener pinning, and hence influence the rheology and growth of the mixed layer. As the grains in the mixed layer shrink, subsequently spalled new grains are also smaller, causing a feedback that leads to more rapid mixing and shear localization in the mixed layer. The early stages of mixing can be compared to laboratory

  12. Effects of Gravity on Bubble Formation at a Plate Orifice

    NASA Technical Reports Server (NTRS)

    Webbon, Bruce W.; Buyevich, Yu A.

    1995-01-01

    A model of the dynamic regime of gas injection through a submerged plate orifice into an ideally wetting liquid is developed in the circumstance when successively detached bubbles may be regarded as independent objects. Two major factors favor bubble detachment: the buoyancy force and a force due to the momentum inflow into the bubble with injected gas. In normal and moderately reduced gravity, the first factor dominates. At relatively low flow rates, a growing bubble is modeled as a spherical segment touching the orifice perimeter during the whole period of its evolution till detachment. If the flow rate exceeds a critical value, another stage of bubble evolution occurs in which an almost spherical gas envelope is connected with the orifice by a nearly cylindrical gaseous stem that lengthens as the bubble rises above the plate. The bubble continues to grow until the gas supply through the stem is completely cut off, after which back flow of gas into the stem from the bubble becomes possible. In microgravity, the second factor prevails, and the latter stage is always present irrespective of the flow rate. However, the gas envelope rises and the stem lengthens very slowly. This difference in the underlying physical mechanisms provides for key properties of bubble growth and detachment being drastically different in appreciable and sufficiently reduced gravity. The frequency of bubble formation slightly decreases with and the detachment volume is almost proportional to the gas flow rate in the first case, in accordance with familiar relations. In the second case, the first variable is proportional to the flow rate whereas the second one is independent of it. Effects of other parameters, such as the orifice radius, gas and liquid densities, and surface tension are discussed.

  13. Influence of spatial effects on coherent structures formation in a supersonic flow near a plate

    NASA Astrophysics Data System (ADS)

    Lipatov, I. I.; Tugazakov, R. Ya.

    2017-06-01

    Formation of coherent structures in supersonic flow near flat plate with a reflecting strong shock wave is studied. Also, a problem of harmonic wave influencing the flow near plate has been investigated. Similar mechanisms of coherent structures formation in both unsteady problems are found due to secondary instability arising in a flow.

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

    PubMed

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

    2003-03-26

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

  15. Signaling pathways and tissue interactions in neural plate border formation.

    PubMed

    Schille, Carolin; Schambony, Alexandra

    2017-01-01

    The neural crest is a transient cell population that gives rise to various cell types of multiple tissues and organs in the vertebrate embryo. Neural crest cells arise from the neural plate border, a region localized at the lateral borders of the prospective neural plate. Temporally and spatially coordinated interaction with the adjacent tissues, the non-neural ectoderm, the neural plate and the prospective dorsolateral mesoderm, is required for neural plate border specification. Signaling molecules, namely BMP, Wnt and FGF ligands and corresponding antagonists are derived from these tissues and interact to induce the expression of neural plate border specific genes. The present mini-review focuses on the current understanding of how the NPB territory is formed and accentuates the need for coordinated interaction of BMP and Wnt signaling pathways and precise tissue communication that are required for the definition of the prospective NC in the competent ectoderm.

  16. Membranes and membrane plates used in ELISPOT.

    PubMed

    Weiss, Alan J

    2005-01-01

    Membrane-bottomed, 96-well plates constitute the format in which the overwhelming majority of enzyme-linked immunospot (ELISPOT) assays are performed. The membranes in these plates are made from either nitrocellulose or polyvinylidene fluoride. These membranes are well suited for ELISPOT because they have high antibody binding capacities and because their white color provides an excellent backdrop for ELISPOT enumeration. These two membranes and, ultimately, the 96-well plates used in ELISPOT assays were commercialized for filtration applications and later optimized for deoxyribonucleic acid hybridization and protein chemistry applications. In this chapter, an overview of the development and biotechnology applications of nitrocellulose and polyvinylidene fluoride membrane is provided and characteristics and attributes of each of the membranes that are relevant to ELISPOT are summarized.

  17. Titer-plate formatted continuous flow thermal reactors: Design and performance of a nanoliter reactor

    PubMed Central

    Chen, Pin-Chuan; Park, Daniel S.; You, Byoung-Hee; Kim, Namwon; Park, Taehyun; Soper, Steven A.; Nikitopoulos, Dimitris E.; Murphy, Michael C.

    2010-01-01

    velocity, fragment length, and copy number. A 99 bp DNA fragment was successfully amplified at flow velocities from 1 mm/s to 3 mm/s, requiring from 8.16 minutes for 20 cycles (24.48 s/cycle) to 2.72 minutes for 20 cycles (8.16 s/cycle), respectively. Yield compared to the same amplification sequence performed using a bench top thermal cycler decreased nonlinearly from 73% (at 1 mm/s) to 13% (at 3 mm/s) with shorter residence time at the optimal temperatures for the reactions due to increased flow rate primarily responsible. Six different DNA fragments with lengths between 99 bp and 997 bp were successfully amplified at 1 mm/s. Repeatable, successful amplification of a 99 bp fragment was achieved with a minimum of 8000 copies of the DNA template. This is the first demonstration and characterization of continuous flow thermal reactors within the 8 mm × 8 mm footprint of a 96-well micro-titer plate and is the smallest continuous flow PCR to date. PMID:20871807

  18. Titer-plate formatted continuous flow thermal reactors: Design and performance of a nanoliter reactor.

    PubMed

    Chen, Pin-Chuan; Park, Daniel S; You, Byoung-Hee; Kim, Namwon; Park, Taehyun; Soper, Steven A; Nikitopoulos, Dimitris E; Murphy, Michael C

    2010-08-06

    velocity, fragment length, and copy number. A 99 bp DNA fragment was successfully amplified at flow velocities from 1 mm/s to 3 mm/s, requiring from 8.16 minutes for 20 cycles (24.48 s/cycle) to 2.72 minutes for 20 cycles (8.16 s/cycle), respectively. Yield compared to the same amplification sequence performed using a bench top thermal cycler decreased nonlinearly from 73% (at 1 mm/s) to 13% (at 3 mm/s) with shorter residence time at the optimal temperatures for the reactions due to increased flow rate primarily responsible. Six different DNA fragments with lengths between 99 bp and 997 bp were successfully amplified at 1 mm/s. Repeatable, successful amplification of a 99 bp fragment was achieved with a minimum of 8000 copies of the DNA template. This is the first demonstration and characterization of continuous flow thermal reactors within the 8 mm × 8 mm footprint of a 96-well micro-titer plate and is the smallest continuous flow PCR to date.

  19. The pisiform growth plate is lost in humans and supports a role for Hox in growth plate formation

    PubMed Central

    Kjosness, Kelsey M; Hines, Jasmine E; Lovejoy, C Owen; Reno, Philip L

    2014-01-01

    The human pisiform is a small, nodular, although functionally significant, bone of the wrist. In most other mammals, including apes and Australopithecus afarensis, pisiforms are elongate. An underappreciated fact is that the typical mammalian pisiform forms from two ossification centers. We hypothesize that: (i) the presence of a secondary ossification center in mammalian pisiforms indicates the existence of a growth plate; and (ii) human pisiform reduction results from growth plate loss. To address these hypotheses, we surveyed African ape pisiform ossification and confirmed the presence of a late-forming secondary ossification center in chimpanzees and gorillas. Identification of the initial ossification center occurs substantially earlier in apes relative to humans, raising questions concerning the homology of the human pisiform and the two mammalian ossification centers. Second, we conducted histological and immunohistochemical analyses of pisiform ossification in mice. We confirm the presence of two ossification centers separated by organized columnar and hypertrophic chondrocyte zones. Flattened chondrocytes were highly mitotic, indicating the presence of a growth plate. Hox genes have been proposed to play a fundamental role in growth plate patterning. The existence of a pisiform growth plate presents an interesting test case for the association between Hox expression and growth plate formation, and could explain the severe effects on the pisiform observed in Hoxa11 and Hoxd11 knockout mice. Consistent with this hypothesis, we show that Hoxd11 is expressed adjacent to the pisiform in late-stage embryonic mouse limbs supporting a role for Hox genes in growth plate specification. This raises questions concerning the mechanisms regulating Hox expression in the developing carpus. PMID:25279687

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

    PubMed

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

    2013-07-01

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

  1. 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

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

    PubMed

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

    2014-02-20

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

  3. Spray Formation during the Impact of a Flat Plate on Water Surface

    NASA Astrophysics Data System (ADS)

    Wang, An; Duncan, James H.

    2015-11-01

    Spray formation during the impact of a flat plate on a water surface is studied experimentally. The plate is mounted on a two-axis carriage that can slam the plate vertically into the water surface as the carriage moves horizontally along a towing tank. The plate is 122 cm by 38 cm and oriented with adjustable pitch and roll angle. The port (lower) edge of the plate is positioned with a 3-mm gap from one of the tank walls. A laser sheet is created in a plane oriented perpendicular to the axis of the horizontal motion of the carriage. The temporal evolution of the spray within the light sheet is measured with a cinematic laser induced fluorescence technique at a frame rate of 800 Hz. Experiments are performed with a fixed plate trajectory in a vertical plane, undertaken at various speeds. Two types of spray are found when the plate has nonzero pitch and roll angles. The first type is composed of a cloud of high-speed droplets and ligaments generated as the port edge of the plate hits the water surface during the initial impact. The second type is a thin sheet of water that grows from the starboard edge of the plate as it moves below the local water level. The geometrical features of the spray are found to be dramatically affected by the impact velocity. The support of the Office of Naval Research under grant N000141310587 is gratefully acknowledged.

  4. Development of a new microtiter plate format for clinically relevant assays.

    PubMed

    Piletska, Elena V; Piletsky, Stanislav S; Whitcombe, Michael J; Chianella, Iva; Piletsky, Sergey A

    2012-02-21

    A new format for the microtiter plate-based assays was proposed. The novelty involves the use of disk-shaped inserts for immobilization of biological and chemical reagents. The internal opening of the disks allows measurements of the reactions by standard microtiter plate readers without any additional steps involving liquid handling. Ideally the plate end-users just have to add the sample and take the measurement without any need of multiple reagent additions or transfer of the liquid to a different plate. The novel assay format also allows handling of reagents which are not soluble in an aqueous environment. As a proof of concept we describe here several model reactions which are compatible with microtiter plate format, such as monitoring enzymatic reactions catalyzed by glucose oxidase (GOx) and urease, measurements of proteins by BCA assay, analysis of pH, and concentration of antioxidants. The "mix and match" approach in the disk-shape format allows multiplexing and could be particularly useful for high throughput screening. One of the potential application areas for this novel assay format could be in a multianalyte system for measurement of clinically relevant analytes in primary care.

  5. A study of the fluorescence measurement using a 96-well microplate by a remodelled parallel luminescent measuring system.

    PubMed

    Kumae, T

    1999-01-01

    To develop an index to measure alveolar macrophage activity, the fluorescent technique for detection of calcium flux was paid special attention. In this study, a parallel luminescence measuring system was remodelled for fluorescence measurement using a 96-well microplate. The fluorescence indicators widely used to measure cytosolic free calcium ion concentration require excitation at ultraviolet (UV) wavelengths. Instruments to produce UV wavelengths are expensive compared to those used to produce visible wavelengths, and UV wavelengths are potentially injurious to cells. To avoid these problems, Fluo 3 (excitation wavelength in the visible range) was used as the fluorescent dye for detecting calcium ions. The parallel luminometer was remodelled successfully for fluorescent measurement as assessed by the results obtained from the measurements of a common fluorescent dye, fluorescein. Concentrations of free calcium ions were measured using Fluo 3 at 37 degrees C to consider the measurement of calcium flux in living cells. Although a linear relationship between concentrations of free calcium ions and fluorescence were observed, a diminution of fluorescence over time was also observed. To measure calcium flux in living cells, further instrumental and experimental improvements are thus needed. Copyright 1999 John Wiley & Sons, Ltd.

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

    PubMed Central

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

    2012-01-01

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

  7. Arabidopsis R-SNARE Proteins VAMP721 and VAMP722 Are Required for Cell Plate Formation

    PubMed Central

    Liu, Peng; Hao, Huaiqing; Jin, Jing Bo; Lin, Jinxing

    2011-01-01

    Background Cell plate formation during plant cytokinesis is facilitated by SNARE complex-mediated vesicle fusion at the cell-division plane. However, our knowledge regarding R-SNARE components of membrane fusion machinery for cell plate formation remains quite limited. Methodology/Principal Findings We report the in vivo function of Arabidopsis VAMP721 and VAMP722, two closely sequence-related R-SNAREs, in cell plate formation. Double homozygous vamp721vamp722 mutant seedlings showed lethal dwarf phenotypes and were characterized by rudimentary roots, cotyledons and hypocotyls. Furthermore, cell wall stubs and incomplete cytokinesis were frequently observed in vamp721vamp722 seedlings. Confocal images revealed that green fluorescent protein-tagged VAMP721 and VAMP722 were preferentially localized to the expanding cell plates in dividing cells. Drug treatments and co-localization analyses demonstrated that punctuate organelles labeled with VAMP721 and VAMP722 represented early endosomes overlapped with VHA-a1-labeled TGN, which were distinct from Golgi stacks and prevacuolar compartments. In addition, protein traffic to the plasma membrane, but not to the vacuole, was severely disrupted in vamp721vamp722 seedlings by subcellular localization of marker proteins. Conclusion/Significance These observations suggest that VAMP721 and VAMP722 are involved in secretory trafficking to the plasma membrane via TGN/early endosomal compartment, which contributes substantially to cell plate formation during plant cytokinesis. PMID:22022536

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

    PubMed

    Nilsson, Karin; Andersson, Maria; Beck, Olof

    2014-11-01

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

  9. Vortex Formation, Shedding and Energy Harvesting from a Cyber-Physical Pitching Flat Plate

    NASA Astrophysics Data System (ADS)

    Onoue, Kyohei; Breuer, Kenneth

    2014-11-01

    We examine the dynamics and energy harvesting capabilities of an elastically mounted flat plate undergoing large amplitude limit cycle oscillations in a uniform flow. All experiments are performed using a cyber-physical system, wherein the structural inertia, stiffness and damping are numerically simulated using a position-following feedback algorithm. The cyber-physical system also allows for implementation of nonlinear spring and damping coefficients, which control the plate dynamics and subsequent energy harvesting characteristics. Analysis of the plate kinematics and the fluid flow over the plate and in the wake (measured using PIV) are used to understand the interplay between structural motion and vortex formation at the sharp leading and trailing edges of the plate. By varying the structural properties of the system we systematically analyze the formation, strength, stability and separation of the leading edge vortex, as well as the dependence on kinematic parameters and Reynolds number. Connections to previous results on vortex formation time and bluff body aerodynamics are discussed. This research is funded by the Air Force Office of Scientific Research (AFOSR).

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

    PubMed

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

    2006-06-07

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

  11. ESET histone methyltransferase is essential to hypertrophic differentiation of growth plate chondrocytes and formation of epiphyseal plates.

    PubMed

    Yang, Liu; Lawson, Kevin A; Teteak, Colin J; Zou, Junhui; Hacquebord, Jacques; Patterson, David; Ghatan, Andrew C; Mei, Qi; Zielinska-Kwiatkowska, Anna; Bain, Steven D; Fernandes, Russell J; Chansky, Howard A

    2013-08-01

    The ESET (also called SETDB1) protein contains an N-terminal tudor domain that mediates protein-protein interactions and a C-terminal SET domain that catalyzes methylation of histone H3 at lysine 9. We report here that ESET protein is transiently upregulated in prehypertrophic chondrocytes in newborn mice. To investigate the in vivo effects of ESET on chondrocyte differentiation, we generated conditional knockout mice to specifically eliminate the catalytic SET domain of ESET protein only in mesenchymal cells. Such deletion of the ESET gene caused acceleration of chondrocyte hypertrophy in both embryos and young animals, depleting chondrocytes that are otherwise available to form epiphyseal plates for endochondral bone growth. ESET-deficient mice are thus characterized by defective long bone growth and trabecular bone formation. To understand the underlying mechanism for ESET regulation of chondrocytes, we carried out co-expression experiments and found that ESET associates with histone deacetylase 4 to bind and inhibit the activity of Runx2, a hypertrophy-promoting transcription factor. Repression of Runx2-mediated gene transactivation by ESET is dependent on its H3-K9 methyltransferase activity as well as its associated histone deacetylase activity. In addition, knockout of ESET is associated with repression of Indian hedgehog gene in pre- and early hypertrophic chondrocytes. Together, these results provide clear evidence that ESET controls hypertrophic differentiation of growth plate chondrocytes and endochondral ossification during embryogenesis and postnatal development.

  12. Spatio-temporal analysis of cellulose synthesis during cell plate formation in Arabidopsis.

    PubMed

    Miart, Fabien; Desprez, Thierry; Biot, Eric; Morin, Halima; Belcram, Katia; Höfte, Herman; Gonneau, Martine; Vernhettes, Samantha

    2014-01-01

    During cytokinesis a new crosswall is rapidly laid down. This process involves the formation at the cell equator of a tubulo-vesicular membrane network (TVN). This TVN evolves into a tubular network (TN) and a planar fenestrated sheet, which extends at its periphery before fusing to the mother cell wall. The role of cell wall polymers in cell plate assembly is poorly understood. We used specific stains and GFP-labelled cellulose synthases (CESAs) to show that cellulose, as well as three distinct CESAs, accumulated in the cell plate already at the TVN stage. This early presence suggests that cellulose is extruded into the tubular membrane structures of the TVN. Co-localisation studies using GFP-CESAs suggest the delivery of cellulose synthase complexes (CSCs) to the cell plate via phragmoplast-associated vesicles. In the more mature TN part of the cell plate, we observed delivery of GFP-CESA from doughnut-shaped organelles, presumably Golgi bodies. During the conversion of the TN into a planar fenestrated sheet, the GFP-CESA density diminished, whereas GFP-CESA levels remained high in the TVN zone at the periphery of the expanding cell plate. We observed retrieval of GFP-CESA in clathrin-containing structures from the central zone of the cell plate and from the plasma membrane of the mother cell, which may contribute to the recycling of CESAs to the peripheral growth zone of the cell plate. These observations, together with mutant phenotypes of cellulose-deficient mutants and pharmacological experiments, suggest a key role for cellulose synthesis already at early stages of cell plate assembly.

  13. Flows, strains, and the formation of joints in oblique collision of metal plates

    NASA Astrophysics Data System (ADS)

    Shtertser, A. A.; Zlobin, B. S.

    2015-09-01

    The processes of high-velocity oblique collision of metal plates which lead to the formation of their joints (seizure) are considered. It is found that the cleaning of the plate surface necessary for seizure results from a jet flow (particle stream), whose source is at least one of the welded materials or an interlayer of ductile material located in the initial region of collision. It is shown that additional cleaning may occur due to the emergence of rotating microregions in intense gradient flows localized in the joint area; seizure on cleaned surfaces is due to reduction of the surface energy of the system.

  14. Novel large format sealed tube microchannel plate detectors for Cherenkov timing and imaging

    NASA Astrophysics Data System (ADS)

    Siegmund, O. H. W.; McPhate, J. B.; Vallerga, J. V.; Tremsin, A. S.; Jelinsky, S. R.; Frisch, H. J.; Lappd Collaboration

    2011-05-01

    Large area (20×20 cm 2) sealed tube detectors using novel borosilicate glass microchannel plates, with bialkali photocathodes and strip-line readouts are being developed for Cherenkov light detection. Designs based on conventional sealed tubes with alumina brazed body construction and hot indium seals have been developed. Borosilicate glass substrates with 20 and 40 μm holes have been processed using atomic layer deposition to produce functional microchannel plates. Initial results for these in a 33 mm format show gain, imaging performance, pulse shape and lifetime characteristics that are similar to standard glass microchannel plates. Large area (20×20 cm 2) borosilicate glass substrates with 20 μm pores have also been made.

  15. SOXC Transcription Factors Induce Cartilage Growth Plate Formation in Mouse Embryos by Promoting Noncanonical WNT Signaling.

    PubMed

    Kato, Kenji; Bhattaram, Pallavi; Penzo-Méndez, Alfredo; Gadi, Abhilash; Lefebvre, Véronique

    2015-09-01

    Growth plates are specialized cartilage structures that ensure the elongation of most skeletal primordia during vertebrate development. They are made by chondrocytes that proliferate in longitudinal columns and then progress in a staggered manner towards prehypertrophic, hypertrophic and terminal maturation. Complex molecular networks control the formation and activity of growth plates, but remain incompletely understood. We investigated here the importance of the SoxC genes, which encode the SOX4, SOX11 and SOX12 transcription factors, in growth plates. We show that the three genes are expressed robustly in perichondrocytes and weakly in growth plate chondrocytes. SoxC(Prx1Cre) mice, which deleted SoxC genes in limb bud skeletogenic mesenchyme, were born with tiny appendicular cartilage primordia because of failure to form growth plates. In contrast, SoxC(Col2Cre) and SoxC(ATC) mice, which deleted SoxC genes primarily in chondrocytes, were born with mild dwarfism and fair growth plates. Chondrocytes in the latter mutants matured normally, but formed irregular columns, proliferated slowly and died ectopically. Asymmetric distribution of VANGL2 was defective in both SoxC(Prx1Cre) and SoxC(ATC) chondrocytes, indicating impairment of planar cell polarity, a noncanonical WNT signaling pathway that controls growth plate chondrocyte alignment, proliferation and survival. Accordingly, SoxC genes were necessary in perichondrocytes for expression of Wnt5a, which encodes a noncanonical WNT ligand required for growth plate formation, and in chondrocytes and perichondrocytes for expression of Fzd3 and Csnk1e, which encode a WNT receptor and casein kinase-1 subunit mediating planar cell polarity, respectively. Reflecting the differential strengths of the SOXC protein transactivation domains, SOX11 was more powerful than SOX4, and SOX12 interfered with the activity of SOX4 and SOX11. Altogether, these findings provide novel insights into the molecular regulation of skeletal

  16. Protective nitride formation on stainless steel alloys for proton exchange membrane fuel cell bipolar plates

    SciTech Connect

    Yang, Bing; Brady, Michael P; Wang, Heli; Turner, John; More, Karren Leslie; Young, David J; Tortorelli, Peter F; Payzant, E Andrew; Walker, Larry R

    2007-01-01

    Gas nitridation has shown excellent promise to form dense, electrically conductive and corrosion-resistant Cr-nitride surface layers on Ni-Cr base alloys for use as proton exchange membrane fuel cell (PEMFC) bipolar plates. Due to the high cost of nickel, Fe-base bipolar plate alloys are needed to meet the cost targets for many PEMFC applications. Unfortunately, nitridation of Fe-base stainless steel alloys typically leads to internal Cr-nitride precipitation rather than the desired protective surface nitride layer formation, due to the high permeability of nitrogen in these alloys. This paper reports the finding that it is possible to form a continuous, protective Cr-nitride (CrN and Cr{sub 2}N) surface layer through nitridation of Fe-base stainless steel alloys. The key to form a protective Cr-nitride surface layer was found to be the initial formation of oxide during nitridation, which prevented the internal nitridation typically observed for these alloys, and resulted in external Cr-nitride layer formation. The addition of V to the alloy, which resulted in the initial formation of V{sub 2}O{sub 3}-Cr{sub 2}O{sub 3}, was found to enhance this effect, by making the initially formed oxide more amenable to subsequent nitridation. The Cr-nitride surface layer formed on model V-modified Fe-27Cr alloys exhibited excellent corrosion resistance and low interfacial contact resistance under simulated PEMFC bipolar plate conditions.

  17. Protective nitride formation on stainless steel alloys for proton exchange membrane fuel cell bipolar plates

    SciTech Connect

    Yang, B.; Brady, M. P.; Wang, H.; Turner, J. A.; More, K. L.; Young, D. J.; Tortorelli, P. F.; Payzant, E. A.; Walker, L. R.

    2007-09-07

    Gas nitridation has shown excellent promise to form dense, electrically conductive and corrosion-resistant Cr-nitride surface layers on Ni–Cr base alloys for use as proton exchange membrane fuel cell (PEMFC) bipolar plates. Due to the high cost of nickel, Fe-base bipolar plate alloys are needed to meet the cost targets for many PEMFC applications. Unfortunately, nitridation of Fe-base stainless steel alloys typically leads to internal Cr-nitride precipitation rather than the desired protective surface nitride layer formation, due to the high permeability of nitrogen in these alloys. This paper reports the finding that it is possible to form a continuous, protective Cr-nitride (CrN and Cr2N) surface layer through nitridation of Fe-base stainless steel alloys. The key to form a protective Cr-nitride surface layer was found to be the initial formation of oxide during nitridation, which prevented the internal nitridation typically observed for these alloys, and resulted in external Cr-nitride layer formation. The addition of V to the alloy, which resulted in the initial formation of V2O3–Cr2O3, was found to enhance this effect, by making the initially formed oxide more amenable to subsequent nitridation. The Cr-nitride surface layer formed on model V-modified Fe–27Cr alloys exhibited excellent corrosion resistance and low interfacial contact resistance under simulated PEMFC bipolar plate conditions.

  18. Mixing at the microscale: Power input in shaken microtiter plates.

    PubMed

    Dürauer, Astrid; Hobiger, Stefanie; Walther, Cornelia; Jungbauer, Alois

    2016-12-01

    Power input and local energy dissipation are crucial parameters for the engineering characterization of mixing and fluid dynamics at the microscale. Since hydrodynamic stress is solely dependent on the maximum power input, we adapted the clay/polymer method to obtain flock destruction kinetics in six-, 24-, and 96-well microtiter plates on orbital shakers. We also determined the specific power input using calorimetry and found that the power input is at the same order of magnitude for the six- and 96-well plates and the laboratory-scale stirred tank reactor, with 40 to 90 W/m(3) (Re' = 180 to 440), 40 to 140 W/m3 (Re' = 320 to 640), and 30 to 50 W/m(3) (Re = 4000 to 8500), respectively. All of these values are significantly below 450 to 2100 W/m(3) determined for the pilot-scale reactor. The hydrodynamic stress differs significantly between the different formats of MTPs, as the 96-well plates showed very low shear stress on the shaker with a shaking amplitude of 3 mm. Thus, the transfer of mixing conditions from the microtiter plate to small-scale and pilot-scale reactors must be undertaken with care. Our findings, especially the power input determined by the calorimetric method, show that the hydrodynamic conditions in laboratory- and pilot-scale reactors cannot be reached. © 2016 The Authors. Biotechnology Journal published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

    PubMed

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

    2004-01-01

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

  20. Growth plate formation and development in alligator and mouse metapodials: evolutionary and functional implications.

    PubMed

    Reno, Philip L; Horton, Walter E; Elsey, Ruth M; Lovejoy, C Owen

    2007-05-15

    Mammalian metapodials (metacarpals and metatarsals), unlike most long bones, form a single growth plate, and undergo longitudinal growth at only one end. The growth dynamics of non-mammalian tetrapod metapodials have not been systematically examined in order to determine if unidirectional growth is unique to mammals. Here we compare murine metapodial ossification in growth stages that parallel those of embryonic, juvenile and subadult American alligators (Alligator mississippiensis). Safranin O staining was used for qualitative histology, and chondrocyte differentiation and proliferation were assessed via immunohistochemistry for type X collagen and proliferative cell nuclear antigen (PCNA). We establish that growth plates form at both ends of alligator metapodials and are maintained in the subadult. PCNA results show that alligators and mice share common patterns of chondrocyte proliferation during growth plate formation. In addition, while alligators and mice differ initially in the degree of organization and pace of chondrocyte differentiation, these parameters are largely similar in established growth plates. However, the replacement of cartilage by bone is highly irregular throughout growth in the alligator, in contrast to the more uniform process in the mouse. These results indicate that while alligators and mammals share common mechanisms of chondrocyte regulation, they differ substantially in their processes of ossification. Phylogenetic analysis indicates that the direct ossification of one epiphysis and reliance on a single growth plate is a derived character (synapomorphy) in therian mammals and likely indicates an adaptation for erect quadrupedal gait.

  1. Folate receptor alpha is necessary for neural plate cell apical constriction during Xenopus neural tube formation.

    PubMed

    Balashova, Olga A; Visina, Olesya; Borodinsky, Laura N

    2017-03-02

    Folate supplementation prevents up to 70% of neural tube defects (NTDs), which result from a failure of neural tube closure during embryogenesis. The elucidation of the mechanisms underlying folate action has been challenging. This study introduces Xenopus laevis as a model to determine the cellular and molecular mechanisms involved in folate action during neural tube formation. We show that knockdown of folate receptor-α (FRα) impairs neural tube formation and leads to NTDs. FRα knockdown in neural plate cells only is necessary and sufficient to induce NTDs. FRα-deficient neural plate cells fail to constrict, resulting in widening of the neural plate midline and defective neural tube closure. Pharmacological inhibition of folate action by methotrexate during neurulation induces NTDs by inhibiting folate interaction with its uptake systems. Our findings support a model for folate receptor interacting with cell adhesion molecules, thus regulating apical cell membrane remodeling and cytoskeletal dynamics necessary for neural plate folding. Further studies in this organism may unveil novel cellular and molecular events mediated by folate and lead to new means for preventing NTDs.

  2. Changes in Pacific Absolute Plate Motion and Formation of Oceanic Flood Basalt Plateaus

    NASA Astrophysics Data System (ADS)

    Kroenke, L. W.; Wessel, P.

    2006-12-01

    The origin of the large oceanic flood basalt plateaus that are prominent features of the central western Pacific Basin remains unclear. Major changes in Pacific Absolute Plate Motion (APM) have been identified as occurring at 145, 125, 96, and 47 Ma. Formation of the Shatsky Rise (~145 Ma), the Ontong Java Plateau (122+ Ma), the Southern Hess Rise (95±5 Ma), and the Louisiade Plateau (~48 Ma) appear to coincide with these changes. A smaller, but still prominent change in Pacific APM also occurred at 110 Ma when the Northern Hess Rise formed. Although these concurrent events may simply be chance occurrences, initiation of plate tectonic reorganizations upon arrival of mantle plume heads also was proposed by Ratcliff et al. (1998), who suggested that the mantle plume head delivery of hot material to produce flood basalts also had the potential to trigger reorganizations of plate motions. It should be noted, however, that Pacific Rim subduction zone development also coincides with these APM changes, and that the actual cause and effect of each change in APM has yet to be clearly established. Here we present a modified Pacific APM model that uses several older seamount chains (Musicians, Ratak-Gilbert-Ellice, the Wake trails, and the Liliuokalani trails) to constrain the oldest Pacific plate motion using the hybrid technique of Wessel et al (2006).

  3. Insight into characteristic features of cartilage growth plate as a physiological template for bone formation.

    PubMed

    Jaroszewicz, Jakub; Kosowska, Anna; Hutmacher, Dietmar; Swieszkowski, Wojciech; Moskalewski, Stanisław

    2016-02-01

    Cartilage growth plate is a natural template from both a biochemical and structural point of view and allows osteoblasts migration, proliferation, differentiation, and ultimately, bone formation. It is evolutionary adjusted to support bone formation within strictly defined spatial framework serving as an interesting model for studying more mechanistically aspects which might be important for specific scaffold-based bone tissue engineering strategies. Surprisingly little is known about the geometric features of this physiological template. To this purpose we analyzed cartilage growth plate from rat, mouse, and human costochondral junction and tibia. High-resolution X-ray tomography showed that pore size in the zone of provisional calcification was within 20 to 30 µm range and in the metaphysis in 35 to 50 µm range. The thickness of calcified longitudinal septa in zone of provisional calcification was 3 to 5 µm and in metaphysis 7 to 12 µm. The porosity varied from 84 to 88%. We observed that numerical values characteristic for cartilage growth plate were not significantly influenced by the species of origin, by the type of bone, or by age. In addition, electron microscopy of calcified fragments of longitudinal septa showed that the calcium aggregates were globular, connected with each other, and formed a shell covering cartilage matrix located within longitudinal septa.

  4. Evaporation-reducing Culture Condition Increases the Reproducibility of Multicellular Spheroid Formation in Microtiter Plates.

    PubMed

    Das, Viswanath; Fürst, Tomáš; Gurská, Soňa; Džubák, Petr; Hajdúch, Marián

    2017-03-07

    Tumor models that closely imitate in vivo conditions are becoming increasingly popular in drug discovery and development for the screening of potential anti-cancer drugs. Multicellular tumor spheroids (MCTSes) effectively mimic the physiological conditions of solid tumors, making them excellent in vitro models for lead optimization and target validation. Out of the various techniques available for MCTS culture, the liquid-overlay method on agarose is one of the most inexpensive methods for MCTS generation. However, the reliable transfer of MCTS cultures using liquid-overlay for high-throughput screening may be compromised by a number of limitations, including the coating of microtiter plates (MPs) with agarose and the irreproducibility of uniform MCTS formation across wells. MPs are significantly prone to edge effects that result from excessive evaporation of medium from the exterior of the plate, preventing the use of the entire plate for drug tests. This manuscript provides detailed technical improvements to the liquid-overlay technique to increase the scalability and reproducibility of uniform MCTS formation. Additionally, details on a simple, semi-automatic, and universally applicable software tool for the evaluation of MCTS features after drug treatment is presented.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    PubMed

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

    2006-02-02

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

  7. Spinodal instability and pattern formation in thin liquid films confined between two plates.

    PubMed

    Verma, Ruhi; Sharma, Ashutosh; Banerjee, Indrani; Kargupta, Kajari

    2006-04-01

    The instability, morphology and pattern formation engendered by the van der Waals force in a thin liquid film of thickness h confined between two closely placed solid surfaces (at distance d > h) are investigated based on nonlinear 3D simulations. The initial and the final stages of dewetting and pattern formation are found to be crucially dependent on the volumetric (thickness) ratio of air and liquid and its deviation from the location of the maximum of the spinodal parameter versus volumetric ratio curve. On a low energy surface, relatively thinner films and wider air gaps favor initial dewetting of the lower plate by the formation of holes, whereas thicker films with thinner air gaps initially evolve by the formation of columns/bridges that join the upper plate. In the later stage of evolution, the initial holes in thinner films evolve into columns/drops, while a rapid coalescence of columns in the thicker films eventually causes formation of holes. Thus, a phase inversion, either from liquid-in-air to air-in-liquid dispersion or vice versa, occurs during the final stages of evolution. A thin film confined between two high-energy solid surfaces forms columns (bridges) only when its mean thickness, h0, is greater than a critical thickness (hc) or the air gap is smaller than a critical distance. The patterns can be aligned by using a topographically patterned confining surface. Conditions on pattern periodicity, amplitude, and the volumetric ratio of air and liquid in the gap are explored for the formation of various types of ordered patterns including annular rings of columns, concentric ripples, parallel channels and a rectangular array of complex features. The results are of significance in soft lithographies such as LISA, soft stamping and capillary force lithography.

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

    USDA-ARS?s Scientific Manuscript database

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

  9. The use of curvilinear fiber format to improve buckling resistance of plates with central circular holes

    NASA Technical Reports Server (NTRS)

    Lee, H. H.; Hyer, M. W.

    1990-01-01

    The potential gain in buckling resistance is explored by using the curvilinear format with a plate containing a centrally located circular hole and subjected to a uniaxial compressive load. The buckling loads for a uniform stress loading and simple support boundary conditions were examined. The method of analysis is outlined for the buckling improvement study. Several designs that represent an improvement over a baseline quasi-isotropic design (a design that is quite good for buckling resistance) are discussed. The tensile load of these designs is examined. A recommended design is presented that not only shows an improved buckling capacity, but also shows improved tensile performance. Follow-on activities are also discussed.

  10. Reproducibility of Uniform Spheroid Formation in 384-Well Plates: The Effect of Medium Evaporation.

    PubMed

    Das, Viswanath; Fürst, Tomáš; Gurská, Soňa; Džubák, Petr; Hajdúch, Marián

    2016-10-01

    Spheroid cultures of cancer cells reproduce the spatial dimension-induced in vivo tumor traits more effectively than the conventional two-dimensional cell cultures. With growing interest in spheroids for high-throughput screening (HTS) assays, there is an increasing demand for cost-effective miniaturization of reproducible spheroids in microtiter plates (MPs). However, well-to-well variability in spheroid size, shape, and growth is a frequently encountered problem with almost every culture method that has prevented the transfer of spheroids to the HTS platform. This variability partly arises due to increased susceptibility of MPs to edge effects and evaporation-induced changes in the growth of spheroids. In this study, we examined the effect of evaporation on the reproducibility of spheroids of tumor and nontumor cell lines in 384-well plates, and show that culture conditions that prevent evaporation-induced medium loss result in the formation of uniform spheroids across the plate. Additionally, we also present a few technical improvements to increase the scalability of the liquid-overlay spheroid culturing technique in MPs, together with a simple software routine for the quantification of spheroid size. We believe that these cost-effective improvements will aid in further improvement of spheroid cultures for HTS drug discovery.

  11. A model for the formation of defects in RPC bakelite plates at high radiation levels

    NASA Astrophysics Data System (ADS)

    Greci, T.; Felli, F.; Saviano, G.; Benussi, L.; Bianco, S.; Passamonti, L.; Piccolo, D.; Pierluigi, D.; Russo, A.

    2013-04-01

    This study analyzes in detail the defects in bakelite observed in Resistive Plate Counters (RPC) after exposure to high-radiation environment and fluxed with humidified gas mixture at 9 kV voltage. Objective of this study was to identify the nature of defects and their formation mechanism. The defects were observed firstly on the whole RPC inner surface, and their localization mapped. The defected areas have been analyzed with optical and electron microscopy (SEM), and chemically by EDS (Energy Dispersion Spectroscopy) techniques. An area particularly defect-rich also analysed by x-ray diffraction (XRD). Samples of new and fluxed bakelite have been chemically analyzed by ICP-Plasma (via sample total digestion) in order to determine trace elements variations in composition. model is proposed to explain the chemistry of the formation process.

  12. Calculating formation range of binary amorphous alloys fabricated by electroless plating

    NASA Astrophysics Data System (ADS)

    Zhang, Bangwei; Liao, Shuzhi; Shu, Xiaolin; Xie, Haowen

    2016-06-01

    A lot of amorphous alloy deposits in the binary (Ni, Co, Cu)-(P, B) alloy systems fabricated by electroless plating (EP) have been reported up to date. But no one reported their theoretical modeling of the amorphous formation and calculated their concentration range of amorphous formation (RAF). Using Miedema model and subregular model scheme, the RAFs for the six EP (Ni, Co, Cu)-(P, B) alloys and three Ni-Cu, Ni-Co and Co-Cu alloys have been calculated systematically for the first time. The calculated results are in agreement with experimental observations. Experiments and calculations for the RAFs in the latter three alloy systems reveal that not any RAF formed except crystalline states. The huge difference between the six metal-metalloid alloys and three metal-metal alloys in RAF has been discussed in detail in the paper.

  13. Mosaic wedge-and-strip arrays for large format microchannel plate detectors

    NASA Technical Reports Server (NTRS)

    Martin, Christopher; Rasmussen, Andrew

    1989-01-01

    The authors present a novel method for joining wedge-and-strip patterns on single anodes in a mosaic array. With only a modest increase in complexity over three-conductor anodes currently in use, the ultimate detector position resolution can be significantly improved, and large-format microchannel plate detectors with pore-size-limited resolution are made possible. The problem of the transition from one anode to the next has been solved with a novel linear encoding scheme, which exhibits essentially distortionless behavior at boundaries parallel to the conducting elements and only slight distortion at the orthogonal boundaries. The ultimate resolution for two anode designs, one designed for large-format imaging and the other for high-resolution spectroscopy, is also predicted.

  14. Mosaic wedge-and-strip arrays for large format microchannel plate detectors

    NASA Technical Reports Server (NTRS)

    Martin, Christopher; Rasmussen, Andrew

    1989-01-01

    The authors present a novel method for joining wedge-and-strip patterns on single anodes in a mosaic array. With only a modest increase in complexity over three-conductor anodes currently in use, the ultimate detector position resolution can be significantly improved, and large-format microchannel plate detectors with pore-size-limited resolution are made possible. The problem of the transition from one anode to the next has been solved with a novel linear encoding scheme, which exhibits essentially distortionless behavior at boundaries parallel to the conducting elements and only slight distortion at the orthogonal boundaries. The ultimate resolution for two anode designs, one designed for large-format imaging and the other for high-resolution spectroscopy, is also predicted.

  15. A transient analysis of frost formation on a parallel plate evaporator

    SciTech Connect

    Martinez-Frias, J.; Aceves, S.M.; Hernandez-Guerrero, A.

    1996-12-31

    This paper presents the development of a transient model for evaluating frost formation on a parallel plate evaporator for heat pump applications. The model treats the frost layer as a porous substance, and applies the equations of conservation of mass, momentum and energy to calculate the growth and densification of the frost layer. Empirical correlations for thermal conductivity and tortuosity as a function of density are incorporated from previous studies. Frost growth is calculated as a function of time, Reynolds number, longitudinal location, plate temperature, and ambient air temperature and humidity. The main assumptions are: ideal gas behavior for air and water vapor, uniform frost density and thermal conductivity across the thickness of the frost layer; and quasi-steady conditions during the whole process. The mathematical model is validated by comparing the predicted values of frost thickness and frost density with results obtained in recent experimental studies. A good agreement was obtained in the comparison. The frost formation model calculates pressure drop and heat transfer resistance that result from the existence of the frost layer, and it can therefore be incorporated into a heat pump model to evaluate performance losses due to frosting as a function of weather conditions and time of operation since the last evaporator defrost.

  16. Formation of Island Arc-Trench System due to Plate Subduction on the Basis of Elastic Dislocation Theory

    NASA Astrophysics Data System (ADS)

    Fukahata, Y.; Matsu'ura, M.

    2015-12-01

    The most conspicuous cumulative deformation in subduction zones is the formation of island arc-trench system. A pair of anomalies in topography and free-air gravity, high in the arc and low around the trench, is observed without exceptions all over the world. Since the 1960s, elastic dislocation theory has been widely used to interpret coseismic crustal deformation. For the modeling of longer-term crustal deformation, it is necessary to consider viscoelastic properties of the asthenosphere. By simply applying elastic-viscoelastic dislocation theory to plate subduction, Matsu'ura and Sato (1989, GJI) have shown that some crustal deformation remains after the completion of one earthquake cycle, which means that crustal deformation accumulates with time in a long term due to plate subduction. In fact, by constructing a plate interface model in and around Japan, Hashimoto, Fukui and Matsu'ura (2004, PAGEOPH) have demonstrated that the computed vertical displacements due to steady plate subduction well explain the observed free-air gravity anomaly pattern. Recently, we got a lucid explanation of crustal deformation due to plate subduction. In subduction zones, oceanic plates bend and descend into the mantle. Because the bending of oceanic plates is usually not spontaneous, there exists kinematic interaction between the oceanic and overriding plates, which causes cumulative deformation of the overriding plate. This may be understood based on the law of action and reaction: one is bending of an oceanic plate and the other is deformation of the overriding plate. As a special case, it is useful to consider plate subduction along a part of true circle. In this case, crustal deformation due to steady subduction is solely caused by the effect of gravity, because dislocation along a circle does not cause any intrinsic internal deformation. When an oceanic plate is descending along an arcuate plate interface from the right-hand side, according to dislocation theory, the oceanic

  17. Influences of surface hydrophilicity on frost formation on a vertical cold plate under natural convection conditions

    SciTech Connect

    Liu, Zhongliang; Zhang, Xinghua; Wang, Hongyan; Meng, Sheng; Cheng, Shuiyuan

    2007-07-15

    Surface hydrophilicity has a strong influence on frost nucleation according to phase transition theory. To study this effect, a close observation of frost formation and deposition processes on a vertical plate was made under free convection conditions. The formation and shape variation of frost crystals during the initial period are described and the frost thickness variation with time on both hydrophobic and plain copper cold surfaces are presented. The various influencing factors are discussed in depth. The mechanism of surface hydrophilicity influence on frost formation was analyzed theoretically. This revealed that increasing the contact angle can increase the potential barrier and restrain crystal nucleation and growth and thus frost deposition. The experimental results show that the initial water drops formed on a hydrophobic surface are smaller and remain in the liquid state for a longer time compared with ones formed on a plain copper surface. It is also observed that the frost layer deposited on a hydrophobic surface is loose and weak. Though the hydrophobic surface can retard frost formation to a certain extent and causes a looser frost layer, our experimental results show that it does not depress the growth of the frost layer. (author)

  18. Effect of formation temperature on properties of graphite/stannum composite for bipolar plate

    SciTech Connect

    Selamat, Mohd Zulkefli Yusuf, Muhammad Yusri Md Wer, Tio Kok Sahadan, Siti Norbaya Malingam, Sivakumar Dhar Mohamad, Noraiham

    2016-03-29

    Bipolar plates are key components in Proton Exchange Membrane (PEM) fuel cells. They carry current away from the cell and withstand the clamping force of the stack assembly. Therefore, PEM fuel cell bipolar plates must have high electrical conductivity and adequate mechanical strength, in addition to being light weight and low cost in terms of both applicable materials and production methods. In this research, the raw materials used to fabricate the high performance bipolar plate are Graphite (Gr), Stannum (Sn) and Polypropylene (PP). All materials used was in powder form and Gr and Sn act as fillers and the PP acts as binder. The ratio of fillers (Gr/Sn) and binder (PP) was fixed at 80:20. For the multi-conductive filler, small amount of Sn, which is 10 up to 20wt% (from the total weight of fillers 80%) have been added into Gr/Sn/PP composite. The fillers were mixed by using the ball mill machine. The second stage of mixing process between the mixer of fillers and binder is also carried out by using ball mill machine before the compaction process by the hot press machine. The effect of formation temperatures (160°C-170°C) on the properties of Gr/Sn/PP composite had been studied in detail, especially the electrical conductivity, bulk density, hardness and microstructure analysis of Gr/Sn/PP composite. The result shows that there are significant improvement in the electrical conductivity and bulk density, which are exceeding the US-DoE target with the maximum value of 265.35 S/cm and 1.682g/cm{sup 3} respectively.

  19. Effect of formation temperature on properties of graphite/stannum composite for bipolar plate

    NASA Astrophysics Data System (ADS)

    Selamat, Mohd Zulkefli; Yusuf, Muhammad Yusri Md; Wer, Tio Kok; Sahadan, Siti Norbaya; Malingam, Sivakumar Dhar; Mohamad, Noraiham

    2016-03-01

    Bipolar plates are key components in Proton Exchange Membrane (PEM) fuel cells. They carry current away from the cell and withstand the clamping force of the stack assembly. Therefore, PEM fuel cell bipolar plates must have high electrical conductivity and adequate mechanical strength, in addition to being light weight and low cost in terms of both applicable materials and production methods. In this research, the raw materials used to fabricate the high performance bipolar plate are Graphite (Gr), Stannum (Sn) and Polypropylene (PP). All materials used was in powder form and Gr and Sn act as fillers and the PP acts as binder. The ratio of fillers (Gr/Sn) and binder (PP) was fixed at 80:20. For the multi-conductive filler, small amount of Sn, which is 10 up to 20wt% (from the total weight of fillers 80%) have been added into Gr/Sn/PP composite. The fillers were mixed by using the ball mill machine. The second stage of mixing process between the mixer of fillers and binder is also carried out by using ball mill machine before the compaction process by the hot press machine. The effect of formation temperatures (160°C-170°C) on the properties of Gr/Sn/PP composite had been studied in detail, especially the electrical conductivity, bulk density, hardness and microstructure analysis of Gr/Sn/PP composite. The result shows that there are significant improvement in the electrical conductivity and bulk density, which are exceeding the US-DoE target with the maximum value of 265.35 S/cm and 1.682g/cm3 respectively.

  20. Enhancement of measurement sensitivity in the formation of shear interferograms of transparent plates with small residual wedging

    SciTech Connect

    But', A I; Lyalikov, A M

    2011-10-31

    We have proposed a method for increasing the sensitivity of measurements of the wedge angle in transparent plates. The method is based on formation of the holographic shear interferograms using a combination of 180 Degree-Sign rotation of the plate with byturn adjustment of interferograms in its images to an infinitely wide fringe. The sensitivity enhancement is due to the increased number of interference fringes in the observed images of the wedged plate, which favours the reduction of the measurement error during optical processing of the obtained interferograms. Data on the experimental validation of the proposed method are presented.

  1. 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...

  2. Validation and application of a 96-well format solid-phase extraction and liquid chromatography-tandem mass spectrometry method for the quantitation of digoxin in human plasma.

    PubMed

    Hashimoto, Yoshitaka; Shibakawa, Kimio; Nakade, Susumu; Miyata, Yasuyuki

    2008-06-15

    To evaluate the pharmacokinetics of digoxin in humans, a sensitive and specific LC/MS/MS method was developed and validated for the determination of digoxin concentrations in human plasma. The method was shown to be more sensitive, specific, accurate, and reproducible than common techniques such as RIA. For detection, a LC/MS/MS system with electro spray ionization tandem mass spectrometry in the positive ion-multiple reaction-monitoring (MRM) mode was used to monitor precursor to product ions of m/z 798.5-51.5 for digoxin and m/z 782.5-35.5 for the internal standard, digitoxin. The method was validated over a concentration range of 0.02-5 ng/mL and was found to have acceptable accuracy, precision, linearity, and selectivity. The mean extraction recovery from spiked plasma samples was above 80%. Imidafenacin, coadministered in a drug-drug interaction study, had no detectable influence on the determination of digoxin in human plasma. The novel method was applied to a drug-drug interaction study of digoxin and imidafenacin and the characterization of steady-state pharmacokinetics of digoxin in humans after oral administration at a dose of 0.25 mg on days 1 and 2 followed by 0.125 mg daily doses on days 3 through 8.

  3. 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...

  4. Overview of membranes and membrane plates used in research and diagnostic ELISPOT assays.

    PubMed

    Weiss, Alan J

    2012-01-01

    Polyvinylidene fluoride (PVDF) membrane-bottomed, 96-well plates and 8-well strips constitute the formats in which the overwhelming majority of ELISPOT assays used in research and diagnostic applications are performed. PVDF is well suited for ELISPOT because it has a high antibody-binding capacity and because its white color provides an excellent backdrop for ELISPOT enumeration. Nitrocellulose (NC) and PVDF membranes and 96-well plates containing those membranes used in ELISPOT assays were initially commercialized for filtration applications and later optimized for a range of different protein analytical applications. An overview of the development and biotechnology applications of PVDF membrane is provided. Characteristics and attributes of the membrane that are relevant to ELISPOT are summarized. Enhancements in PVDF membrane performance and optimization of devices for automation compatible and diagnostic ELISPOT applications are presented.

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

    PubMed

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

    2005-01-05

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

  6. Geodynamical Nature of the Formation of Large Plates of Platforms, Jointed in North Caspian Oil and Gas Basin

    ERIC Educational Resources Information Center

    Seitov, Nassipkali; Tulegenova, Gulmira P.

    2016-01-01

    This article addresses the problems of tectonic zoning and determination of geodynamical nature of the formation of jointed tectonic structures within the North Caspian oil and gas basin, represented by Caspian Depression of Russian platform of East European Pre-Cambrian Craton and plate ancient Precambrian Platform stabilization and Turan…

  7. Arc/Forearc Lengthening at Plate Triple Junctions and the Formation of Ophiolitic Soles

    NASA Astrophysics Data System (ADS)

    Casey, John; Dewey, John

    2013-04-01

    The principal enigma of large obducted ophiolite slabs is that they clearly must have been generated by some form of organized sea-floor spreading/plate-accretion, such as may be envisioned for the oceanic ridges, yet the volcanics commonly have arc affinity (Miyashiro) with boninites (high-temperature/low-pressure, high Mg and Si andesites), which are suggestive of a forearc origin. PT conditions under which boninites and metamorphic soles form and observations of modern forearc systems lead us to the conclusion that ophiolite formation is associated with overidding plate spreading centers that intersect the trench to form ridge-trench-trench of ridge-trench-tranform triple junctions. The spreading centers extend and lengthen the forearc parallel to the trench and by definition are in supra-subduction zone (SSZ) settings. Many ophiolites likewise have complexly-deformed associated mafic-ultramafic assemblages that suggest fracture zone/transform t along their frontal edges, which in turn has led to models involving the nucleation of subduction zones on fracture zones or transpressional transforms. Hitherto, arc-related sea-floor-spreading has been considered to be either pre-arc (fore-arc boninites) or post-arc (classic Karig-style back arc basins that trench-parallell split arcs). Syn-arc boninites and forearc oceanic spreading centers that involve a stable ridge/trench/trench triple or a ridge-trench-transform triple junction, the ridge being between the two upper plates, are consistent with large slab ophiolite formation in a readied obduction settting. The direction of subduction must be oblique with a different sense in the two subduction zones and the oblique subduction cannot be partitioned into trench orthogonal and parallel strike-slip components. As the ridge spreads, new oceanic lithosphere is created within the forearc, the arc and fore-arc lengthen significantly, and a syn-arc ophiolite forearc complex is generated by this mechanism. The ophiolite

  8. Paleocene Pacific Plate reorganization mirrored in formation of the Suvarov Trough, Manihiki Plateau

    NASA Astrophysics Data System (ADS)

    Pietsch, Ricarda; Uenzelmann-Neben, Gabriele

    2016-10-01

    The Suvarov Trough is a graben structure that deviates from the Danger Islands Troughs within the Manihiki Plateau, a Large Igneous Province (LIP) located in the Central Pacific. New high-resolution seismic reflection data provide evidence that the graben formed in two phases during the Paleocene (65-45 Ma). In a first phase extension occurred in southwestward direction, pulling apart the northern part of the Suvarov Trough and a parallel trending unnamed trough. In a second phase a change of extensional force direction occurred from southwest to west-northwest, forming the southern part of the Suvarov Trough that extends onto the High Plateau. The formation of the Suvarov Trough is accompanied by a series of normal fault systems that apparently formed simultaneously. Comparing the seismic results to existing Pacific paleo strain reconstructions, the timing of increased strain and local deformation direction fits well to our findings. We thus suggest that the multiple strike directions of the Suvarov Trough represent an extensional structure that was caused by the major, stepwise Pacific Plate reorganization during the Paleocene.

  9. Surface topography formation in a region of plate collision: Mathematical modeling

    NASA Astrophysics Data System (ADS)

    Korobeinikov, S. N.; Reverdatto, V. V.; Polyanskii, O. P.; Sverdlova, V. G.; Babichev, A. V.

    2012-07-01

    The collision of earth's crustal plates is modeled mathematically based on a numerical solution of the equations of deformable solid mechanics using a finite element method with the MSC software. The interaction of the plates with each other and with the mantle is described by the solution of the contact problem with an unknown contact boundary between the solids considered. The mantle material is assumed to be ideal elastic-plastic with the Huber-Mises yield surface, and the properties of the plate material are described using an elastic-plastic model with the Drucker-Prager parabolic yield function which takes into account fracture in the tensile stress region. The results of the mathematical modeling show that the surface profiles of the plates in the region of their collision are consistent, both qualitatively and quantitatively, to the surface topography observed in nature under similar conditions.

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

    PubMed

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

    2016-11-02

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

  11. One-step synthesis of collagen hybrid gold nanoparticles and formation on Egyptian-like gold-plated archaeological ivory.

    PubMed

    Spadavecchia, Jolanda; Apchain, Emilande; Albéric, Marie; Fontan, Elisabeth; Reiche, Ina

    2014-08-04

    A one-step method is reported to synthesize hybrid gold nanoparticles (AuNPs) by reduction of HAuCl4 in acetic solution in the presence of collagen (Col), dicarboxylic acid-terminated polyethylene glycol (PEG), and cetyltetrammonium bromide (CTAB) mixed with hydoxyapatite (HAP) as surfactants. Such formation process of AuNPs was shown to be responsible for purple stains naturally formed on Egyptianizing archaeological gilded ivories from 8th BC Syria. The understanding of this formation mechanism, which most likely involves a step with hybrid AuNPs, allows the establishing of an authenticity marker of ancient gold-plated ivories.

  12. Paper microzone plates.

    PubMed

    Carrilho, Emanuel; Phillips, Scott T; Vella, Sarah J; Martinez, Andres W; Whitesides, George M

    2009-08-01

    This paper describes 96- and 384-microzone plates fabricated in paper as alternatives to conventional multiwell plates fabricated in molded polymers. Paper-based plates are functionally related to plastic well plates, but they offer new capabilities. For example, paper-microzone plates are thin (approximately 180 microm), require small volumes of sample (5 microL per zone), and can be manufactured from inexpensive materials ($0.05 per plate). The paper-based plates are fabricated by patterning sheets of paper, using photolithography, into hydrophilic zones surrounded by hydrophobic polymeric barriers. This photolithography used an inexpensive formulation photoresist that allows rapid (approximately 15 min) prototyping of paper-based plates. These plates are compatible with conventional microplate readers for quantitative absorbance and fluorescence measurements. The limit of detection per zone loaded for fluorescence was 125 fmol for fluorescein isothiocyanate-labeled bovine serum albumin, and this level corresponds to 0.02 the quantity of analyte per well used to achieve comparable signal-to-noise in a 96-well plastic plate (using a solution of 25 nM labeled protein). The limits of detection for absorbance on paper was approximately 50 pmol per zone for both Coomassie Brilliant Blue and Amaranth dyes; these values were 0.4 that required for the plastic plate. Demonstration of quantitative colorimetric correlations using a scanner or camera to image the zones and to measure the intensity of color, makes it possible to conduct assays without a microplate reader.

  13. Excimer laser-induced formation of metallic microstructures by electroless copper plating

    NASA Astrophysics Data System (ADS)

    Yang, H.; Pan, C.-T.

    2002-03-01

    Micro-patterns created by the excimer laser and activated by reactants for electroless copper plating are described in this paper. The generated micro-patterns are transformed into copper patterns on the substrate and copper microstructures are formed. This method simplifies the manufacturing process of making circuits on boards compared with the conventional lithography process of forming copper patterns on the substrate. Micro-patterns generated by the excimer laser cause changes of surface electric properties and activation selectively. A chemical reaction through these activated areas may deposit metal, such as copper. The KrF excimer laser not only provides simple and fast machining patterns, but also uses its high-energy density to drill holes and circuits directly. Palladium ions are added as mediators in the electroless plating solution to enable a continuous electroless copper deposition. According to the experiment of excimer laser-assisted electroless copper plating, the procedures of pretreatment and post-cleaning are the key factors that resulted in excellent selective plating. The samples were pretreated by sodium dodecyl sulfate (SDS) and post-cleaned by acetone and diluted nitric acid resulting in distinct micro-patterns. The deposition area is confined to the excimer laser-ablated portion resulting in good selective plating.

  14. The use of curvilinear fiber format to improve buckling resistance of composite plates with central circular holes

    NASA Technical Reports Server (NTRS)

    Hyer, M. W.; Lee, H. H.

    1991-01-01

    The gains in buckling performance are explored that can be achieved by deviating from the conventional straightline fiber format and considering the situation whereby the fiber orientation in a layer, or a group of layers, can vary from point to point. The particular situation studied is a simply supported square plate with a centrally located hole loaded in compression. By using both a sensitivity analysis and a gradient-search technique, fiber orientation in a number of regions of the plate are selected so as to increase the buckling load relative to baseline straightline designs. The sensitivity analysis is used to determine which regions of the plate have the most influence on buckling load, and the gradient search is used to find the design that is believed to represent the absolute maximum buckling load for the conditions prescribed. Convergence studies and sensitivity of the final design are discussed. By examining the stress resultant contours, it is shown how the curvilinear fibers move the load away from the unsupported hole region of the plate to the supported edges, thus increasing the buckling capacity. The tensile capacity of the improved buckling design is investigated, and it is shown that both tensile capacity and buckling capacity can be improved with the curvilinear fiber concept.

  15. Dynamics of phragmoplastin in living cells during cell plate formation and uncoupling of cell elongation from the plane of cell division.

    PubMed Central

    Gu, X; Verma, D P

    1997-01-01

    The cell plate is formed by the fusion of Golgi apparatus-derived vesicles in the center of the phragmoplast during cytokinesis in plant cells. A dynamin-like protein, phragmoplastin, has been isolated and shown to be associated with cell plate formation in soybean by using immunocytochemistry. In this article, we demonstrate that similar to dynamin, phragmoplastin polymerizes to form oligomers. We fused soybean phragmoplastin with the green fluorescence protein (GFP) and introduced it into tobacco BY-2 cells to monitor the dynamics of early events in cell plate formation. We demonstrate that the chimeric protein is functional and targeted to the cell plate during cytokinesis in transgenic cells. GFP-phragmoplastin was found to appear first in the center of the forming cell plate, and as the cell plate grew outward, it redistributed to the growing margins of the cell plate. The redistribution of phragmoplastin may require microtubule reorganization because the microtubule-stabilizing drug taxol inhibited phragmoplastin redistribution. Our data show that throughout the entire process of cytokinesis, phragmoplastin is concentrated in the area in which membrane fusion is active, suggesting that phragmoplastin participates in an early membrane fusion event during cell plate formation. Based on the dynamics of GFP-phragmoplastin, it appears that the process of cell plate formation is completed in two phases. The first phase is confined to the cylinder of the phragmoplast proper and is followed by a second phase that deposits phragmoplast vesicles in a concentric fashion, resulting in a ring of fluorescence, with the concentration of vesicles being higher at the periphery. In addition, overexpression of GFP-phragmoplastin appears to act as a dominant negative, slowing down the completion of cell plate formation, and often results in an oblique cell plate. The latter appears to uncouple cell elongation from the plane of cell division, forming twisted and elongated cells

  16. Development prospects for Bazhenov formation bituminous claystones in the southeast of the West Siberian Plate (Tomsk Region)

    NASA Astrophysics Data System (ADS)

    Belozerov, V. B.; Baranov, V. E.; Dmitriev, A. Y.

    2015-02-01

    The article considers the use of time-tested bituminous shale development technologies in the southeast of the West Siberian Plate (Tomsk Region). A research target is the bituminous claystones of the Bazhenov formation. The Bazhenov formation was divided into stratigraphic units, and the sequence of forming these units within the territory under study was restored on the basis of the peculiarities of the Bazhenov formation section structure with gamma-ray logging data. The joint analysis of logs (gamma-ray logging, resistivity logging, SP logging, neutron gamma-ray logging, induction logging) and core data revealed the wide development of carbonatization processes in the lower stratigraphic units of the Bazhenov formation, whereas the upper, most bituminous units do not have the signs of carbonatization. It is a favorable factor for using the existing technologies of producing hydrocarbons from analogous sedimentary deposits. Considering the degree of lithological disconnection of an oilbearing object from adjacent reservoir formations allows making a conclusion that the application of the existing technologies for developing bituminous claystones can be implemented only in the insignificant territory of the area under study where the Bazhenov formation overlies Georgiev formation claystones being over 8 meters thick. To develop the oil and gas potential of the Bazhenov formation in other territories, there will be a need for new technologies that allow forming a fracture-cavernous matrix in a carbonate rock.

  17. Influence of thick surface coatings on jet formation in flyer plate impacts

    NASA Astrophysics Data System (ADS)

    Georges, William; Loiseau, Jason; Higgins, Andrew; Tyler, Troy; Zimmermann, Joerg

    2017-01-01

    The criteria for jetting upon the impact of flyer plates is that an oblique shock solution in the impact point reference frame does not exist. This criterion has been extensively verified experimentally over the last few decades. Comparatively, there has been no studies in the open literature regarding the jetting criterion for the impact of multi-layered flyers or flyer plates featuring thick coatings. In this work we have developed an experimental set-up that allowed us to determine whether or not the impact of multi-layered flyers resulted in a jet or not using laser heterodyne velocimetry.

  18. Callus Formation and Mineralization after Fracture with Different Fixation Techniques: Minimally Invasive Plate Osteosynthesis versus Open Reduction Internal Fixation

    PubMed Central

    Ding, Haoliang; Qin, Hui; An, Zhiquan

    2015-01-01

    Minimally invasive plate osteosynthesis(MIPO) has been considered as an alternative for fracture treatment. Previous study has demonstrated that MIPO technique has the advantage of less soft tissue injury compared with open reduction internal fixation (ORIF). However, the comparison of callus formation and mineralization between two plate osteosynthesis methods remains unknown. In this experiment, ulna fracture model was established in 42 beagle dogs. The fractures underwent reduction and internal fixation with MIPO or ORIF. Sequential fluorescent labeling and radiographs were applied to determine new callus formation and mineralization in two groups after operation. At 4, 8 and 12 weeks postoperatively, the animals were selected to be sacrificed and the ulna specimens were analyzed by Micro-CT. The sections were also treated with Masson staining for histological evaluation. More callus formation was observed in MIPO group in early stage of fracture healing. The fracture union rate has no significant difference between two groups. The results indicate that excessive soft tissue stripping may impact early callus formation. As MIPO technique can effectively reduce soft tissue injury with little incision, it is considered to be a promising alternative for fracture fixation. PMID:26444295

  19. Optimization of reaction parameters in hydrothermal synthesis: a strategy towards the formation of CuS hexagonal plates

    PubMed Central

    2013-01-01

    Background For decades, copper sulphide has been renowned as the superior optical and semiconductor materials. Its potential applications can be ranged from solar cells, lithium-ion batteries, sensors, and catalyst systems. The synthesis methodologies of copper sulphide with different controlled morphology have been widely explored in the literature. Nevertheless, the understanding on the formation chemistry of CuS is still limited. The ultimate approach undertaking in this article is to investigate the formation of CuS hexagonal plates via the optimization of reaction parameters in hydrothermal reaction between copper (II) nitrate and sodium thiosulphate without appending any assistant agent. Results Covellite (CuS) hexagonal plates were formed at copper ion: thiosulphate ion (Cu2+:S2O32−) mole ratio of 1:2 under hydrothermal treatment of 155°C for 12 hours. For synthesis conducted at reaction temperature lower than 155°C, copper sulphate (CuSO4), krohnite (NaCu2(SO4)(H2O)2] and cyclooctasulphur (S8) were present as main impurities with covellite (CuS). When Cu2+:S2O32− mole ratio was varied to 1: 1 and 1: 1.5, phase pure plate-like natrochalcite [NaCu2(SO4)(H2O)] and digenite (Cu9S5) were produced respectively. Meanwhile, mixed phases of covellite (CuS) and cyclooctasulphur (S8) were both identified when Cu2+:S2O32− mole ratio was varied to 1: 2.5, 1: 3 and 1: 5 as well as when reaction time was shortened to 1 hour. Conclusions CuS hexagonal plates with a mean edge length of 1 μm, thickness of 100 nm and average crystallite size of approximately (45 ± 2) nm (Scherrer estimation) were successfully synthesized via assisting agent- free hydrothermal method. Under a suitable Cu2+:S2O32− mole ratio, we evidenced that the formation of covellite (CuS) is feasible regardless of the reaction temperature applied. However, a series of impurities were attested with CuS if reaction temperature was not elevated high enough for the additional crystallite

  20. Touch-plate and statolith formation in graviceptors of ephyrae which developed while weightless in space

    NASA Technical Reports Server (NTRS)

    Spangenberg, D. B.; Coccaro, E.; Schwarte, R.; Lowe, B.

    1996-01-01

    Ultrastructural studies of the statocysts and touch-plates of graviceptors (rhopalia) of Aurelia ephyrae revealed that (1) touch-plate hair cells are present; and (2) cytoplasmic strands from the hair cell bases extend from the neurite plexus to touch similar strands from the lithocytes. This close association of hair cell neurites and statocysts may have important implications regarding the transmitting and processing of positional information with respect to the gravity vector. Graviceptors of ephyrae which developed while weightless in microgravity were compared with controls at the ultrastructural level. We found that hair cells of ephyrae which developed in microgravity had fewer lipid droplets in the large spaces near their bases as compared with 1 g controls. In the ephyrae from the first microgravity experiment, hair cells had more large apical vacuoles with filamentous content than were found in hair cells of ephyrae from the second experiment and controls. The neurite plexus and the network of cytoplasmic strands extending to the statocysts were not different in microgravity-developed ephyrae from controls. Behavioral differences in swimming and orienting in ephyrae in microgravity and controls (reported earlier) were not explained by morphological differences in the hair cells of the touch-plates or the statocysts, although functional differences apparently occurred.

  1. BEAMS, MODES: Formation of the beam profile in a plate electrode HF laser

    NASA Astrophysics Data System (ADS)

    Andramanov, A. V.; Kabaev, S. A.; Lazhintsev, B. V.; Nor-Arevyan, V. A.; Pisetskaya, A. V.; Selemir, Victor D.

    2005-04-01

    The near-field energy-density distribution of a beam from an HF electric-discharge laser with an inductively stabilised discharge, an SF6- and H2-based active medium, and interelectrode gaps of 34 and 52 mm is studied. Plates 1-mm thick and 60- or 120-mm wide oriented at an angle of 30° to the optical axis serve as 60-mm-wide electrodes. Electrodes 30-mm wide are formed by 60-mm-wide plates also oriented at an angle of 30° to the optical axis. The distributions of the laser-radiation density in a resonator with flat mirrors and the discharge-emission intensity over the resonator cross section are studied. The distributions of the laser-radiation density in the near-field zone and the discharge-emission intensity have similar profiles. The arrangement of the electrode plates at an angle of 30° provided a considerable increase in the width of the lasing region. The FWHM of the laser-radiation distribution is ~80% of the electrode width for the both interelectrode gaps.

  2. Touch-plate and statolith formation in graviceptors of ephyrae which developed while weightless in space

    NASA Technical Reports Server (NTRS)

    Spangenberg, D. B.; Coccaro, E.; Schwarte, R.; Lowe, B.

    1996-01-01

    Ultrastructural studies of the statocysts and touch-plates of graviceptors (rhopalia) of Aurelia ephyrae revealed that (1) touch-plate hair cells are present; and (2) cytoplasmic strands from the hair cell bases extend from the neurite plexus to touch similar strands from the lithocytes. This close association of hair cell neurites and statocysts may have important implications regarding the transmitting and processing of positional information with respect to the gravity vector. Graviceptors of ephyrae which developed while weightless in microgravity were compared with controls at the ultrastructural level. We found that hair cells of ephyrae which developed in microgravity had fewer lipid droplets in the large spaces near their bases as compared with 1 g controls. In the ephyrae from the first microgravity experiment, hair cells had more large apical vacuoles with filamentous content than were found in hair cells of ephyrae from the second experiment and controls. The neurite plexus and the network of cytoplasmic strands extending to the statocysts were not different in microgravity-developed ephyrae from controls. Behavioral differences in swimming and orienting in ephyrae in microgravity and controls (reported earlier) were not explained by morphological differences in the hair cells of the touch-plates or the statocysts, although functional differences apparently occurred.

  3. Effect of Fe- and Si-Enriched Secondary Precipitates and Surface Roughness on Pore Formation on Aluminum Plate Surfaces During Anodizing

    NASA Astrophysics Data System (ADS)

    Zhu, Yuanzhi; Wang, Shizhi; Yang, Qingda; Zhou, Feng

    2014-09-01

    Two twin roll casts (TRCs) and one hot rolled (HR) AA 1235 aluminum alloy plates with different microstructures are prepared. The plates were electrolyzed in a 1.2 wt% HCl solution with a voltage of 21 V and a current of 1.9 mA. The shape, size, and number of pores formed on the surfaces of these plates were analyzed and correlated with the microstructures of the plates. It is found that pores are easier to form on the alloy plates containing subgrains with a lower dislocation density inside the subgrains, rather than along the grain boundaries. Furthermore, Fe- and Si-enriched particles in the AA1235 aluminum alloys lead to the formation of pores on the surface during electrolyzing; the average precipitate sizes of 4, 3.5, and 2 μm in Alloy 1#, Alloy 2# and Alloy 3# result in the average pore sizes of 3.78, 2.76, and 1.9 μm on the surfaces of the three alloys, respectively; The G.P zone in the alloy also facilitates the surface pore formation. High-surface roughness enhances the possibility of entrapping more lubricants into the plate surface, which eventually blocks the formation of the pores on the surface of the aluminum plates in the following electrolyzing process.

  4. A novel automated electrochemical ascorbic acid assay in the 24-well microtiter plate format.

    PubMed

    Intarakamhang, Sireerat; Leson, Christian; Schuhmann, Wolfgang; Schulte, Albert

    2011-02-14

    Automatic ascorbic acid (AA) voltammetry was established in 24-well microtiter plates. The assay used a movable assembly of a pencil rod working, an Ag/AgCl reference and a Pt counter electrode with differential pulse voltammetry (DPV) for concentration-dependent current generation. A computer was in command of electrode (z) and microtiter plate (x, y) positioning and timed potentiostat operation. Synchronization of these actions supported sequential approach of all wells and subsequent execution of electrode treatment procedures or AA voltammetry at defined intervals in a measuring cycle. DPV in well solutions offered a linear current/concentration range between 0.1 and 8.0 mM, a sensitivity of about 1 μA mM(-1) AA, and a detection limit of 50 μM. When used with a calibration curve or standard addition, automated voltammetry of samples with added known amounts of AA demonstrated good recovery rates. Also, the assay achieved the accurate determination of the AA content of vitamin C tablets, a fruit juice and an herbal tea extract. Robotic AA voltammetry has the advantage of conveniently handling multiple samples in a single measuring run without the continuous attention of laboratory personnel. It is a good option when the goal is cost-effective AA screening of sample libraries and has potential for applications in health care and the food processing, cosmetic and pharmaceutical industries.

  5. Plate interface rheological switches during subduction infancy: Control on slab penetration and metamorphic sole formation

    NASA Astrophysics Data System (ADS)

    Agard, P.; Yamato, P.; Soret, M.; Prigent, C.; Guillot, S.; Plunder, A.; Dubacq, B.; Chauvet, A.; Monié, P.

    2016-10-01

    Subduction infancy corresponds to the first few million years following subduction initiation, when slabs start their descent into the mantle. It coincides with the transient (yet systematic) transfer of material from the top of the slab to the upper plate, as witnessed by metamorphic soles welded beneath obducted ophiolites. Combining structure-lithology-pressure-temperature-time data from metamorphic soles with flow laws derived from experimental rock mechanics, this study highlights two main successive rheological switches across the subduction interface (mantle wedge vs. basalts, then mantle wedge vs. sediments; at ∼800 °C and ∼600 °C, respectively), during which interplate mechanical coupling is maximized by the existence of transiently similar rheologies across the plate contact. We propose that these rheological switches hinder slab penetration and are responsible for slicing the top of the slab and welding crustal pieces (high- then low-temperature metamorphic soles) to the base of the mantle wedge during subduction infancy. This mechanism has implications for the rheological properties of the crust and mantle (and for transient episodes of accretion/exhumation of HP-LT rocks in mature subduction systems) and highlights the role of fluids in enabling subduction to overcome the early resistance to slab penetration.

  6. Periodic acid-Schiff's reagent assay for carbohydrates in a microtiter plate format.

    PubMed

    Kilcoyne, Michelle; Gerlach, Jared Q; Farrell, Mark P; Bhavanandan, Veer P; Joshi, Lokesh

    2011-09-01

    Microtiter plate colorimetric assays are widely used for analysis of carbohydrates and glycoconjugates. However, mucins are often not easily detected, as they have low neutral sugar content. We have adapted and optimised the periodic acid-Schiff's reagent (PAS) staining for microtiter plate assay by examining five factors: concentration and volume of periodic acid, oxidation time, volume of Schiff's reagent, and color development time. This assay requires just 25 μl of sample, utilises standardised Schiff's reagent, and has decreased assay time (140 min to completion). Seventeen monosaccharides (acidic, neutral, basic, phosphorylated, and deoxy) and four disaccharides were assessed. PAS-positive carbohydrates (amino, N-acetylamino, deoxy, and certain neutral monosaccharides, and sialic acids) responded linearly within a 10-100 nmol range approximately, which varied for each carbohydrate. The assay response for fetuin and porcine gastric mucin (PGM) was linear up to 150 μg (highest concentration tested), with no response from nonglycosylated protein. A lower response for asialofetuin was observed, but desialylated PGM preparations were similar or higher in response than their sialylated counterparts. The simplicity and low sample consumption of this method make it an excellent choice for screening or quantitation of chromatographic fractions containing carbohydrates and glycoconjugates, especially in the case of mucins. Copyright © 2011 Elsevier Inc. All rights reserved.

  7. Exhumation and Coupling at the Plate Interface: Large Tectonic Slices V. Melange Formation? Key Contexts and Possible Controlling Parameters

    NASA Astrophysics Data System (ADS)

    Agard, P.; Angiboust, S.; Guillot, S.; Garcia-Casco, A.

    2011-12-01

    Fragments of subducted oceanic lithosphere returned along the plate interface convey crucial information regarding the thermal and rheological conditions of convergent plate boundaries. Geological evidence indicate that, unlike subduction, exhumation is non-steady (Agard et al., Earth Sci. Rev. 2009). We herein focus on deep processes along the plate interface (40-80 km depth), for which there is no counterflow (unlike in accretionary prisms) and no other known mechanisms to return eclogites than interplate friction or buoyancy. These eclogites are of two major types: large scale (>km) slices with coherent PT estimates (W. Alps) versus isolated fragments (frequently m-hm) in a serpentinite- or sedimentary-rich matrix showing contrasting equilibration depths (with hints of punctuated exhumation and even reburial; Franciscan, Cuba, Sistan; e.g., Garcia-Casco et al., Geol. Acta 2006). This latter type tends to show warmer equilibration paths, whereas the larger tectonic slices from the former type remain systematically cold. Serpentinites are crucial for both in permitting decoupling and acting as a buoy, and fluid budget is important too in enhancing floatability and allowing large slices to survive (Angiboust and Agard, Lithos 2010). Numerical models implementing free migration of fluids in the subduction zone also show that the plate interface is strongly localized in the absence of fluids: mechanical decoupling efficiently occurs along the sediment veneer and/or at the top of the highly hydrothermalized crust. Whenever fluids are released in greater amounts (depending on initial fluid content and/or thermal structure), deformation becomes much more distributed and affects both the mantle wedge and the top of the downgoing lithosphere (crust and hydrated mantle top), thereby increasing mechanical coupling between the two plates. Based on natural data and numerical modelling we herein propose that rheological contrast chiefly controls mechanical decoupling. On a

  8. Inhibition of Candida albicans CC biofilms formation in polystyrene plate surfaces by biosurfactant produced by Trichosporon montevideense CLOA72.

    PubMed

    Monteiro, Andrea S; Miranda, Tatiana T; Lula, Ivana; Denadai, Ângelo M L; Sinisterra, Rubén D; Santoro, Marcelo M; Santos, Vera L

    2011-06-01

    This study evaluated the effects of glycolipid-type biosurfactant produced by Trichosporon montevideense CLOA72 in the formation of biofilms in polystyrene plate surfaces by Candida albicans CC isolated from the apical tooth canal. Biofilm formation was reduced up to 87.4% with use of biosurfactant at 16 mg/ml concentration. It has been suggested that the interaction with the cell or polystyrene plate surface could ultimately be responsible for these actions. Therefore, the interaction of C. albicans CC cells with the biosurfactant, as well as the corresponding thermodynamic parameters, have been determined by isothermal titration calorimetry and zeta potential measurements. This process is endothermic (((int)H°=+1284±5 cal/mg OD(600)) occurring with a high increase of entropy (T((int)S°=+10635 cal/mg OD(600)). The caloric energy rate data released during the titulation indicates saturation of the cell-biosurfactant at 1.28 mg/ml OD(600). Also, the zeta potential of the cell surface was monitored as a function of the biosurfactant concentration added to cell suspension showing partial neutralization of net surface charge, since the value of zeta potential ranged from -16 mV to -6 mV during the titration. The changes of cell surface characteristics can contribute to the inhibition of initial adherence of cells of C. albicans in surface. The CMC of the purified biosurfactant produced from T. montevideense CLOA72 is 2.2 mg/ml, as determined both by ITC dilution experiments and by surface tension measurements. This biomolecule did not presented any cytotoxic effect in HEK 293A cell line at concentrations of 0.25-1 mg/ml. This study suggests a possible application of the referred biosurfactant in inhibiting the formation of biofilms on plastic surfaces by C. albicans.

  9. Antibody array in a multiwell plate format for the sensitive and multiplexed detection of important plant pathogens.

    PubMed

    Charlermroj, Ratthaphol; Himananto, Orawan; Seepiban, Channarong; Kumpoosiri, Mallika; Warin, Nuchnard; Gajanandana, Oraprapai; Elliott, Christopher T; Karoonuthaisiri, Nitsara

    2014-07-15

    The global seed market is considered to be an important industry with a total value of $10,543 million US dollars in 2012. Because plant pathogens such as bacteria and viruses cause a significant economic loss to both producers and exporters, the seed export industry urgently requires rapid, sensitive, and inexpensive testing for the pathogens to prevent disease spreading worldwide. This study developed an antibody array in a multiwell plate format to simultaneously detect four crucial plant pathogens, namely, a bacterial fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac), Chilli veinal mottle virus (ChiVMV, potyvirus), Watermelon silver mottle virus (WSMoV, tospovirus serogroup IV), and Melon yellow spot virus (MYSV, tospovirus). The capture antibodies specific to the pathogens were immobilized on each well at preassigned positions by an automatic microarrayer. The antibodies on the arrays specifically captured the corresponding pathogens present in the sample extracts. The presence of pathogens bound on the capture antibodies was subsequently detected by a cocktail of fluorescently conjugated secondary antibodies. The limits of detection of the developed antibody array for the detection of Aac, ChiVMV, WSMoV, and MYSV were 5 × 10(5) CFU/mL, 30 ng/mL, 1000 ng/mL, and 160 ng/mL, respectively, which were very similar to those of the conventional ELISA method. The antibody array in a multiwell plate format accurately detected plant pathogens in single and multiple detections. Moreover, this format enables easy handling of the assay at a higher speed of operation.

  10. Novel 96-well quantitative bioelectrocatalytic analysis platform reveals highly efficient direct electrode regeneration of cytochrome P450 BM3 on indium tin oxide.

    PubMed

    Frank, Ronny; Klenner, Marcus; Azendorf, Ronny; Bartz, Manuel; Jahnke, Heinz-Georg; Robitzki, Andrea A

    2017-07-15

    Enzymes are the most effective catalysts for a broad range of difficult chemical reactions e.g. hydroxylation of non-activated C-H Bonds and stereoselective synthesis. Nevertheless, a lot of enzymes are not accessible for the biotechnological applications or industrial use. One reason is the prerequisite of expensive cofactors. In this context, we developed a bioelectrocatalytic analysis platform for the electrochemical and photonic quantification of the direct electron transfer from the electrode to redox enzymes and therefore, bypass the need of soluble cofactors that had to be continuously exchanged or regenerated. As reference enzyme, we chose cytochrome P450 BM3 that is restricted by NADPH dependence. We optimized the substrate spectrum for aromatic compounds by introduction of the triple mutation A74G/F87V/L188Q and established a sensitive fluorimetric product formation assay to monitor the enzymatic conversion of 7-ethoxycoumarine to 7-hydroxycoumarine. Gold and indium tin oxide electrodes were characterized with respect to surface morphology, charge-transfer resistance and P450 BM3 immobilization as well as activity. Using gold electrodes, no significant product formation by electrode mediated direct electron transfer could be detected. In contrast, P450 BM3 adsorbed on unmodified indium tin oxide electrodes revealed 36% activity by electrode mediated direct electron transfer in comparison to enzyme regeneration by NADPH. Since the reaction volumes are in the microliter range and upscaling of the measurement system is easily possible, our analysis platform is a useful tool for bioelectrocatalytic enzyme characterization and library screening based optimization for applications in the field of enzyme catalyzed chemical synthesis but also enzyme based fuel cells. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. A Cell-based PDE4 Assay in 1536-well Plate format for High Throughput Screening

    PubMed Central

    Titus, Steven A.; Li, Xiao; Southall, Noel; Lu, Jianming; Inglese, James; Brasch, Michael; Austin, Christopher P.; Zheng, Wei

    2009-01-01

    The cyclic nucleotide phosphodiesterases (PDEs) are intracellular enzymes that catalyze the hydrolysis of 3', 5'-cyclic nucleotides, such as cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), to their corresponding 5'-nucleotide monophosphates. These enzymes play an important role in controlling cellular concentrations of cyclic nucleotides and thus regulate a variety of cellular signaling events. PDEs are emerging as drug targets for several diseases including asthma, cardiovascular disease, ADHD, Parkinson’s disease, and Alzheimer’s disease. Though biochemical assays with purified recombinant PDE enzymes and cAMP or cGMP substrate are commonly used for compound screening, cell-based assays would provide a better assessment of compound activity in a more physiological context. Here we report the development and validation of a new cell-based PDE4 assay using a constitutively active GPCR as a driving force for cAMP production and a cyclic nucleotide gated (CNG) cation channel as a biosensor in 1536-well plates. PMID:18591513

  12. Early biofilm formation and the effects of antimicrobial agents on orthodontic bonding materials in a parallel plate flow chamber.

    PubMed

    Chin, Mervyn Y H; Busscher, Henk J; Evans, Robert; Noar, Joseph; Pratten, Jonathan

    2006-02-01

    Decalcification is a commonly recognized complication of orthodontic treatment with fixed appliances. A technology, based on a parallel plate flow chamber, was developed to investigate early biofilm formation of a strain of Streptococcus sanguis on the surface of four orthodontic bonding materials: glass ionomer cement (Ketac Cem), resin-modified glass ionomer cement (Fuji Ortho LC), chemically-cured composite resin (Concise) and light-cured composite resin (Transbond XT). S. sanguis was used as it is one of the primary colonizers of dental hard surfaces. Artificial saliva was supplied as a source of nutrients for the biofilms. The effects of two commercially available mouthrinses (i.e. a fluoride containing rinse and chlorhexidine) were evaluated. Initial colonization of the bacterium was assessed after 6 hours of growth by the percentage surface coverage (PSC) of the biofilm on the disc surfaces. There were statistically significant differences in bacterial accumulation between different bonding materials (P < 0.05), Concise being the least colonized and Transbond XT being the most colonized by S. sanguis biofilms. All materials pre-treated with 0.05 per cent sodium fluoride mouthrinse showed more than 50 per cent reduction in biofilm formation. The 0.2 per cent chlorhexidine gluconate mouthrinse caused significant reduction of biofilm formation on all materials except Ketac Cem. This in vitro study showed that the use of a chemically-cured composite resin (Concise) reduced early S. sanguis biofilm formation. Also, fluoride had a greater effect in reducing the PSC by S. sanguis biofilms than chlorhexidine. Rinsing with 0.05 per cent sodium fluoride prior to placement of orthodontic appliances is effective in reducing early biofilm formation.

  13. A semi-automated 96-well plate method for the simultaneous determination of oral contraceptives concentrations in human plasma using ultra performance liquid chromatography coupled with tandem mass spectrometry.

    PubMed

    Licea-Perez, Hermes; Wang, Sherry; Bowen, Chester L; Yang, Eric

    2007-06-01

    Two semi-automated, relatively high throughput methods using ultra performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) were developed for the simultaneous determination of ethinyl estradiol (EE) in combination with either 19-norethindrone (NE) or levonorgestrel (LN) in human plasma. Using 300 microL plasma, the methods were validated over the concentration ranges of 0.01-2 ng/mL and 0.1-20 ng/mL for EE and NE (or LN), respectively. The existing methods for the determination of the oral contraceptives in human plasma require large volumes of plasma (> or =500 microL), and sample extraction is labor-intensive. The LC run time is at least 6 min, enabling analysis of only about 100 samples a day. In the present work the throughput was greatly improved by employing a semi-automated sample preparation process involving liquid-liquid extraction and derivatization with dansyl chloride followed by UPLC separation on a small particle size column achieving a run time of 2.7 min. The validation and actual sample analysis results show that both methods are rugged, precise, accurate, and well suitable to support pharmacokinetic studies where approximately 300 samples can be extracted and analyzed in a day.

  14. The HigB/HigA Toxin/Antitoxin System of Pseudomonas aeruginosa Influences the Virulence Factors Pyochelin, Pyocyanin, and Biofilm Formation

    DTIC Science & Technology

    2016-08-24

    499 Introduction Pseudomonas aeruginosa is an opportunistic, Gram - negative pathogen (Mace et al. 2008), and it is the primary cause of important...formation was assayed in 96- well polystyrene plates using 0.1% crystal violet staining as described previously (Fletcher 1977) with some modifications...the genome of P. aeruginosa PA14 by using the RASTA Bacteria pro- gram (Sevin and Barloy- Hubler 2007). This program searches for type II TA systems

  15. Thermochemical Constraints For the Formation Conditions of the Hydrothermal Alteration Mineralogy of Home Plate and Columbia Hills

    NASA Astrophysics Data System (ADS)

    Filiberto, J.; Schwenzer, S. P.

    2012-12-01

    Home Plate is a plateau in the Columbia Hills of Gusev Crater. It is dominated by igneous minerals (olivine, pyroxene, and magnetite) with small amounts of alteration minerals (hematite and nanophase oxides). Surrounding Home Plate are deposits containing diverse secondary mineral assemblages: Fe3+-sulfates deposits at Paso Robles, Dead Sea, Shredded, Arad, Tyrone, and Troy; Hematite-rich outcrops between Home Plate and Tyrone; SiO2-rich deposits possibly containing pyrite and/or marcasite at Fuzzy Smith; SiO2-rich, possibly opaline silica, deposits at Northern Valley, Eastern Valley, and Tyrone; and Mg-Fe-carbonate outcrops at Comanche in the Columbia Hills [1-4]. Here, we focus on using thermochemical modeling to understand the secondary alteration mineralogy at the Home Plate outcrop and surrounding Columbia Hills region in Gusev Crater. We use CHILLER [5] to evaluate mineral assemblages that are likely to form from the Martian Home Plate, Barn-Hill class rock Fastball in contact with a dilute fluid at various pressures, temperatures, and water-rock ratios. For details see [6]. In our models, hematite dominates the alteration assemblage at high W/R at 150°C, but is generally produced at W/R above 10. Goethite only forms at low temperature and W/R above 40 with a maximum around 100 and again around 100,000. Pyrite is produced at all temperatures but only at relatively high W/R. These results imply intermediate to high W/R and low to intermediate temperatures during alteration of the Home Plate region. Additional acidic brine, while not strictly excluded, is not required to form many of the observed phases. In contrast, the phyllosilicates recently invoked from orbital observations [4] indicate neutral to alkaline conditions - either accompanying the silica precipitation or as a separate event. For future exploration, our results emphasize that the observation of assemblages is critically important to understand mineral formation conditions and that minor phases

  16. Titer plate formatted continuous flow thermal reactors for high throughput applications: fabrication and testing

    NASA Astrophysics Data System (ADS)

    Sang-Won Park, Daniel; Chen, Pin-Chuan; You, Byoung Hee; Kim, Namwon; Park, Taehyun; Lee, Tae Yoon; Datta, Proyag; Desta, Yohannes; Soper, Steven A.; Nikitopoulos, Dimitris E.; Murphy, Michael C.

    2010-05-01

    A high throughput, multi-well (96) polymerase chain reaction (PCR) platform, based on a continuous flow (CF) mode of operation, was developed. Each CFPCR device was confined to a footprint of 8 × 8 mm2, matching the footprint of a well on a standard micro-titer plate. While several CFPCR devices have been demonstrated, this is the first example of a high-throughput multi-well continuous flow thermal reactor configuration. Verification of the feasibility of the multi-well CFPCR device was carried out at each stage of development from manufacturing to demonstrating sample amplification. The multi-well CFPCR devices were fabricated by micro-replication in polymers, polycarbonate to accommodate the peak temperatures during thermal cycling in this case, using double-sided hot embossing. One side of the substrate contained the thermal reactors and the opposite side was patterned with structures to enhance thermal isolation of the closely packed constant temperature zones. A 99 bp target from a λ-DNA template was successfully amplified in a prototype multi-well CFPCR device with a total reaction time as low as ~5 min at a flow velocity of 3 mm s-1 (15.3 s cycle-1) and a relatively low amplification efficiency compared to a bench-top thermal cycler for a 20-cycle device; reducing the flow velocity to 1 mm s-1 (46.2 s cycle-1) gave a seven-fold improvement in amplification efficiency. Amplification efficiencies increased at all flow velocities for 25-cycle devices with the same configuration.

  17. Formation of Australian continental margin highlands driven by plate-mantle interaction

    NASA Astrophysics Data System (ADS)

    Müller, R. Dietmar; Flament, Nicolas; Matthews, Kara J.; Williams, Simon E.; Gurnis, Michael

    2016-05-01

    Passive margin highlands occur on most continents on Earth and play a critical role in the cycle of weathering, erosion, and atmospheric circulation. Yet, in contrast to the well-developed understanding of collisional mountain belts, such as the Alps and Himalayas, the origin of less elevated (1-2 km) passive margin highlands is still unknown. The eastern Australian highlands are a prime example of these plateaus, but compared to others they have a well-documented episodic uplift history spanning 120 million years. We use a series of mantle convection models to show that the time-dependent interaction of plate motion with mantle downwellings and upwellings accounts for the broad pattern of margin uplift phases. Initial dynamic uplift of 400-600 m from 120-80 Ma was driven by the eastward motion of eastern Australia's margin away from the sinking eastern Gondwana slab, followed by tectonic quiescence to about 60 Ma in the south (Snowy Mountains). Renewed uplift of ∼700 m in the Snowy Mountains is propelled by the gradual motion of the margin over the edge of the large Pacific mantle upwelling. In contrast the northernmost portion of the highlands records continuous uplift from 120 Ma to present-day totalling about 800 m. The northern highlands experienced a continuous history of dynamic uplift, first due to the end of subduction to the east of Australia, then due to moving over a large passive mantle upwelling. In contrast, the southern highlands started interacting with the edge of the large Pacific mantle upwelling ∼ 40- 50 million years later, resulting in a two-phase uplift history. Our results are in agreement with published uplift models derived from river profiles and the Cretaceous sediment influx into the Ceduna sub-basin offshore southeast Australia, reflecting the fundamental link between dynamic uplift, fluvial erosion and depositional pulses in basins distal to passive margin highlands.

  18. Comparison of inclined plate sedimentation and dissolved air flotation for the minimisation of subsequent nitrogenous disinfection by-product formation.

    PubMed

    Chu, Wen-Hai; Gao, Nai-Yun; Templeton, Michael R; Yin, Da-Qiang

    2011-04-01

    The formation of disinfection by-products (DBPs), including both nitrogenous disinfection by-products (N-DBPs) and carbonaceous disinfection by-products (C-DBPs), was investigated upon chlorination of water samples following two treatment processes: (i) coagulation-inclined plate sedimentation (IPS)-filtration and (ii) coagulation-dissolved air flotation (DAF)-filtration. The removal of algae, dissolved organic nitrogen (DON), dissolved organic carbon (DOC) and UV(254) by coagulation-DAF-filtration was superior to coagulation-IPS-filtration. On average, 53%, 53% and 31% of DOC, DON and UV(254) were removed by coagulation-DAF-filtration process, which were higher than 47%, 31% and 27% of that by coagulation-IPS-filtration process. Additionally, coagulation-IPS-filtration performed less well at removing the low molecular weight organics than coagulation-DAF-filtration process. The concentrations of chloroform, dichloroacetamide (DCAcAm) and dichloroacetonitrile (DCAN) formed during chlorination after coagulation-DAF-filtration reached their maximum values of 13, 1.5 and 4.7μgL(-1), respectively, and were lower than those after coagulation-IPS-filtration with the maximum detected levels of 17, 2.9 and 6.3μgL(-1). However, the trichloronitromethane (TCNM) concentration after the two processes was similar, suggesting that DON may have less of a contribution to TCNM formation than DCAcAm and DCAN.

  19. IGF-I Signaling in Osterix-Expressing Cells Regulates Secondary Ossification Center Formation, Growth Plate Maturation, and Metaphyseal Formation During Postnatal Bone Development.

    PubMed

    Wang, Yongmei; Menendez, Alicia; Fong, Chak; ElAlieh, Hashem Z; Kubota, Takuo; Long, Roger; Bikle, Daniel D

    2015-12-01

    To investigate the role of IGF-I signaling in osterix (OSX)-expressing cells in the skeleton, we generated IGF-I receptor (IGF-IR) knockout mice ((OSX)IGF-IRKO) (floxed-IGF-IR mice × OSX promoter-driven GFP-labeled cre-recombinase [(OSX)GFPcre]), and monitored postnatal bone development. At day 2 after birth (P2), (OSX)GFP-cre was highly expressed in the osteoblasts in the bone surface of the metaphysis and in the prehypertrophic chondrocytes (PHCs) and inner layer of perichondral cells (IPCs). From P7, (OSX)GFP-cre was highly expressed in PHCs, IPCs, cartilage canals (CCs), and osteoblasts (OBs) in the epiphyseal secondary ossification center (SOC), but was only slightly expressed in the OBs in the metaphysis. Compared with the control mice, the IPC proliferation was decreased in the (OSX)IGF-IRKOs. In these mice, fewer IPCs invaded into the cartilage, resulting in delayed formation of the CC and SOC. Immunohistochemistry indicated a reduction of vessel number and lower expression of VEGF and ephrin B2 in the IPCs and SOC of (OSX)IGF-IRKOs. Quantitative real-time PCR revealed that the mRNA levels of the matrix degradation markers, MMP-9, 13 and 14, were decreased in the (OSX)IGF-IRKOs compared with the controls. The (OSX)IGF-IRKO also showed irregular morphology of the growth plate and less trabecular bone in the tibia and femur from P7 to 7 weeks, accompanied by decreased chondrocyte proliferation, altered chondrocyte differentiation, and decreased osteoblast differentiation. Our data indicate that during postnatal bone development, IGF-I signaling in OSX-expressing IPCs promotes IPC proliferation and cartilage matrix degradation and increases ephrin B2 production to stimulate vascular endothelial growth factor (VEGF) expression and vascularization. These processes are required for normal CC formation in the establishment of the SOC. Moreover, IGF-I signaling in the OSX-expressing PHC is required for growth plate maturation and osteoblast differentiation in

  20. Metamorphic sole formation and early plate interface rheology: Insights from Griggs apparatus experiments

    NASA Astrophysics Data System (ADS)

    Soret, Mathieu; Agard, Philippe; Dubacq, Benoît; Hirth, Greg; Yamato, Philippe; Ildefonse, Benoît; Prigent, Cécile

    2016-04-01

    800°C with confining pressure of 2 GPa. Another simple-shear experiment has been carried out at 800°C and 1 GPa with fined-grained natural garnet. With the aim of mimicking the early slab interface (between the metamorphic sole and banded peridotites at the base of the ophiolite), 2 simple-shear deformation experiments with 2 layers have been carried out at 800°C and confining pressure of 1 GPa. The bottom layer was made of hydrated basalt powder and the top layer was made of olivine. Fined-grained garnet-free amphibolite is significantly weaker than dunite but the appearance of harder minerals in the amphibolite (garnet and clinopyroxene) has major implications on its rheological evolution. These results allow liking field observations of strain localization at the interface to the metamorphic sole formation.

  1. A kinematic model for the formation of the Siletz terrane by capture of coherent fragments of the Farallon and Resurrection plates (Invited)

    NASA Astrophysics Data System (ADS)

    Wilson, D. S.; McCrory, P. A.

    2010-12-01

    The details of the formation of the mostly Eocene basaltic Siletz terrane in the coast ranges of Oregon and Washington have remained enigmatic. Early models [e.g. Duncan, 1982 JGR], noting younger ages for basalts and overlying foramifera in the center of the terrane near the Columbia River and older ages to the north and south, called for an interpretation of a captured fossil spreading ridge between the Kula and Farallon plates. However, later work showed two problems with this simple model [Wells et al., 1984 Tectonics]. First, the difference between old and young ages was larger than predicted by the relatively fast spreading rate. Second, the presence of proximal continental sediments interbedded with some of the older basalts conflicted with the large amounts of convergence predicted by the generally used models for motion of the Kula and Farallon plates relative to North America. The recent suggestion that an additional oceanic plate, the Resurrection plate, was present between the Kula and Farallon plates and subducting beneath eastern Alaska and western Canada during the early Cenozoic [Haeussler et al., 2003 GSAB] permits important revisions to the interpretation of regional tectonic history. We propose a model in which the older (~55 Ma) parts of the Siletz terrane formed at the Resurrection-Farallon ridge, not far offshore from the Klamath Mountains. Proximity to the Yellowstone hotspot could have generated thick oceanic crust in a setting analogous to Iceland. When the Juan de Fuca (Vancouver) plate separated from the Farallon plate at ~52 Ma, we propose that a small Siletz plate also separated from the northern Juan de Fuca plate, eventually to be captured in the Oregon coast range. A fragment of the Resurrection plate may have separated later (~48 Ma), eventually captured in the Washington coast range and constituting the southward-younging part of the terrane. Cessation of motion of these fragments relative to adjacent parts of North America may

  2. Fuel swelling and interaction layer formation in the SELENIUM Si and ZrN coated U(Mo) dispersion fuel plates irradiated at high power in BR2

    NASA Astrophysics Data System (ADS)

    Leenaers, A.; Van den Berghe, S.; Koonen, E.; Kuzminov, V.; Detavernier, C.

    2015-03-01

    In the framework of the SELENIUM project two full size flat fuel plates were produced with respectively Si and ZrN coated U(Mo) particles and irradiated in the BR2 reactor at SCK•CEN. Non-destructive analysis of the plates showed that the fuel swelling profiles of both SELENIUM plates were very similar to each other and none of the plates showed signs of pillowing or excessive swelling at the end of irradiation at the highest power position (local maximum 70% 235U). The microstructural analysis showed that the Si coated fuel has less interaction phase formation at low burn-up but at the highest burn-ups, defects start to develop on the IL-matrix interface. The ZrN coated fuel, shows a virtual absence of reaction between the U(Mo) and the Al, up to high fission densities after which the interaction layer formation starts and defects develop in the matrix near the U(Mo) particles. It was found and is confirmed by the SELENIUM (Surface Engineering of Low ENrIched Uranium-Molybdenum) experiment that there are two phenomena at play that need to be controlled: the formation of an interaction layer and swelling of the fuel. As the interaction layer formation occurs at the U(Mo)-matrix interface, applying a diffusion barrier (coating) at that interface should prevent the interaction between U(Mo) and the matrix. The U(Mo) swelling, observed to proceed at an accelerating rate with respect to fission density accumulation, is governed by linear solid state swelling and fission gas bubble swelling due to recrystallization of the fuel. The examination of the SELENIUM fuel plates clearly show that for the U(Mo) dispersion fuel to be qualified, the swelling rate at high burn-up needs to be reduced.

  3. Formation of aerial standing wave field using ultrasonic sources consisting of multiple stripe-mode transverse vibrating plates

    NASA Astrophysics Data System (ADS)

    Naito, Koki; Asami, Takuya; Miura, Hikaru

    2015-07-01

    Intense aerial acoustic waves can be produced by an ultrasonic source consisting of a transverse vibrating plate and an external jutting driving point. Previously, we studied the dimensional parameters of vibrating plates to produce stripe-mode patterns and thereby determine the plate dimensions that generate high-quality patterns. In this research, we use four transverse vibrating plates as ultrasonic sources to produce intense standing wave fields in air. As a result, an aerial standing wave field was formed in the field surrounded by four vibrating plates. Furthermore, for a total input power of 30 W for the two ultrasonic sources, a very strong (sound pressure level, 167 dB) wave field is obtained.

  4. 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.

  5. Electron Tomographic Analysis of Somatic Cell Plate Formation in Meristematic Cells of Arabidopsis Preserved by High-Pressure FreezingW⃞

    PubMed Central

    Seguí-Simarro, José M.; Austin, Jotham R.; White, Erin A.; Staehelin, L. Andrew

    2004-01-01

    We have investigated the process of somatic-type cytokinesis in Arabidopsis (Arabidopsis thaliana) meristem cells with a three-dimensional resolution of ∼7 nm by electron tomography of high-pressure frozen/freeze-substituted samples. Our data demonstrate that this process can be divided into four phases: phragmoplast initials, solid phragmoplast, transitional phragmoplast, and ring-shaped phragmoplast. Phragmoplast initials arise from clusters of polar microtubules (MTs) during late anaphase. At their equatorial planes, cell plate assembly sites are formed, consisting of a filamentous ribosome-excluding cell plate assembly matrix (CPAM) and Golgi-derived vesicles. The CPAM, which is found only around growing cell plate regions, is suggested to be responsible for regulating cell plate growth. Virtually all phragmoplast MTs terminate inside the CPAM. This association directs vesicles to the CPAM and thereby to the growing cell plate. Cell plate formation within the CPAM appears to be initiated by the tethering of vesicles by exocyst-like complexes. After vesicle fusion, hourglass-shaped vesicle intermediates are stretched to dumbbells by a mechanism that appears to involve the expansion of dynamin-like springs. This stretching process reduces vesicle volume by ∼50%. At the same time, the lateral expansion of the phragmoplast initials and their CPAMs gives rise to the solid phragmoplast. Later arriving vesicles begin to fuse to the bulbous ends of the dumbbells, giving rise to the tubulo-vesicular membrane network (TVN). During the transitional phragmoplast stage, the CPAM and MTs disassemble and then reform in a peripheral ring phragmoplast configuration. This creates the centrifugally expanding peripheral cell plate growth zone, which leads to cell plate fusion with the cell wall. Simultaneously, the central TVN begins to mature into a tubular network, and ultimately into a planar fenestrated sheet (PFS), through the removal of membrane via clathrin

  6. Critical roles of the TGF-β type I receptor ALK5 in perichondrial formation and function, cartilage integrity, and osteoblast differentiation during growth plate development

    PubMed Central

    Matsunobu, Tomoya; Torigoe, Kiyoyuki; Ishikawa, Masaki; de Vega, Susana; Kulkarni, Ashok B.; Iwamoto, Yukihide; Yamada, Yoshihiko

    2009-01-01

    TGF-β has been implicated in the proliferation and differentiation of chondrocytes and osteoblasts. However, the in vivo function of TGF-β in skeletal development is unclear. In this study, we investigated the role of TGF-β signaling in growth plate development by creating mice with a conditional knockout of the TGF-β type I receptor ALK5 (ALK5CKOCKO) in skeletal progenitor cells using Dermo1-Cre mice. ALK5CKO mice had short and wide long bones, reduced bone collars, and trabecular bones. In ALK5CKO growth plates, chondrocytes proliferated and differentiated, but ectopic cartilaginous tissues protruded into the perichondrium. In normal growth plates, ALK5 protein was strongly expressed in perichondrial progenitor cells for osteoblasts, and in a thin chondrocyte layer located adjacent to the perichondrium in the peripheral cartilage. ALK5CKO growth plates had an abnormally thin perichondrial cell layer and reduced proliferation and differentiation of osteoblasts. These defects in the perichondrium likely caused the short bones and ectopic cartilaginous protrusions. Using tamoxifen-inducible Cre-ER™-mediated ALK5-deficient primary calvarial cell cultures, we found that TGF-β signaling promoted osteoprogenitor proliferation, early differentiation, and commitment to the osteoblastic lineage through the selective MAPKs and Smad2/3 pathways. These results demonstrate the important roles of TGF-β signaling in perichondrium formation and differentiation, as well as in growth plate integrity during skeletal development. PMID:19501582

  7. Induction of multinucleation by beta-glucosyl Yariv reagent in regenerated cells from Marchantia polymorpha protoplasts and involvement of arabinogalactan proteins in cell plate formation.

    PubMed

    Shibaya, Taeko; Sugawara, Yasutake

    2009-08-01

    Arabinogalactan proteins (AGPs) are abundant plant cell surface proteoglycans widely distributed in plant species. Since high concentrations of beta-glucosyl Yariv reagent (betaglcY), which binds selectively to AGPs, inhibited cell division of protoplast-regenerated cells of the liverwort Marchantia polymorpha L. (Shibaya and Sugawara in Physiol Plant 130:271-279, 2007), we investigated the mechanism underlying the inability of the cells to divide normally by staining nuclei, cell walls and beta-1,3-glucan. Microscopic observation showed that the diameter of regenerated cells cultured with betaglcY was about 2.8-fold larger than that of cells cultured without betaglcY. The cells cultured with betaglcY were remarkably multinucleated. These results indicated that betaglcY did not inhibit mitosis but induced multinucleation. In the regenerated cells cultured with low concentrations of betaglcY (5 and 1 microg ml(-1)), the cell plate was stained strongly by betaglcY, suggesting abundant AGPs in the forming cell plate. In these cell plates, beta-1,3-glucan was barely detectable or not detected. In multinucleated cells, cell plate-like fragments, which could not reach the cell wall, were frequently observed and they were also stained strongly by betaglcY. Our results indicated that AGPs might have an important role in cell plate formation, and perturbation of AGPs with betaglcY might result in remarkable multinucleation in protoplast-regenerated cells of M. polymorpha.

  8. Critical roles of the TGF-beta type I receptor ALK5 in perichondrial formation and function, cartilage integrity, and osteoblast differentiation during growth plate development.

    PubMed

    Matsunobu, Tomoya; Torigoe, Kiyoyuki; Ishikawa, Masaki; de Vega, Susana; Kulkarni, Ashok B; Iwamoto, Yukihide; Yamada, Yoshihiko

    2009-08-15

    TGF-beta has been implicated in the proliferation and differentiation of chondrocytes and osteoblasts. However, the in vivo function of TGF-beta in skeletal development is unclear. In this study, we investigated the role of TGF-beta signaling in growth plate development by creating mice with a conditional knockout of the TGF-beta type I receptor ALK5 (ALK5(CKO)) in skeletal progenitor cells using Dermo1-Cre mice. ALK5(CKO) mice had short and wide long bones, reduced bone collars, and trabecular bones. In ALK5(CKO) growth plates, chondrocytes proliferated and differentiated, but ectopic cartilaginous tissues protruded into the perichondrium. In normal growth plates, ALK5 protein was strongly expressed in perichondrial progenitor cells for osteoblasts, and in a thin chondrocyte layer located adjacent to the perichondrium in the peripheral cartilage. ALK5(CKO) growth plates had an abnormally thin perichondrial cell layer and reduced proliferation and differentiation of osteoblasts. These defects in the perichondrium likely caused the short bones and ectopic cartilaginous protrusions. Using tamoxifen-inducible Cre-ER-mediated ALK5-deficient primary calvarial cell cultures, we found that TGF-beta signaling promoted osteoprogenitor proliferation, early differentiation, and commitment to the osteoblastic lineage through the selective MAPKs and Smad2/3 pathways. These results demonstrate the important roles of TGF-beta signaling in perichondrium formation and differentiation, as well as in growth plate integrity during skeletal development.

  9. Compaction history of Upper Cretaceous shale (Al-Khod Formation) and its relationship to plate margin tectonics, Arabian Plate, Sultanate of Oman

    NASA Astrophysics Data System (ADS)

    AL-Sarmi, Musaab; Mattern, Frank; Scharf, Andreas; Pracejus, Bernhard; Al-Mamari, Amira; Al-Hinaai, Al-Shima

    2017-04-01

    Conglomerates of the late Cretaceous Al-Khod Formation have been intruded by older shale of the same formation along faults, which were opened/widened by extension, thus, resulting in shale dike formation. Following intrusion the shale was behaving plastically as its fissility follows the contact contours of the conglomeratic host rock and as stoped sandstone blocks are floating within the shale. Vertical calcite veins were ptygmatically folded with subhorizontal fold axial planes. All these aspects show that the shale contained a high water content in the beginning. The ptygmatically folded calcite veins display vertical shortening amounts of 40 % corresponding to 35 % to 45 % of water loss during compaction. Incalculable numbers of calcite veins of different orientations and thicknesses within the conglomerate along the shale contact indicate that the fluid was expelled from the shale into the conglomerate host rock under high pressure (overpressure?). Shale dyke formation took place after the late Cretaceous obduction of the Semail Ophiolite, before the deposition of early Tertiary carbonate rocks, and during the latest Cretaceous doming of the Saih Hatat area which was caused by deformation and slab breakoff, likely associated with gravitational collapse and elastic rebound. Shale intrusion was followed by deposition of 100 to 200 m thick sediments of the upper part of Al-Khod Formation, leading to compaction and water loss. The shale retained much of its water during the uppermost Cretaceous-late Paleocene stratigraphic hiatus as this interval is marked by erosion and a reduction of overburden, which was probably due to the elastic rebound. Folding of calcite veins together with a high amount of water loss was a consequence of compaction caused by the overburden of 1000 m thick shallow marine limestones which were deposited from the Eocene to Oligocene.

  10. Innovative design of composite structures: The use of curvilinear fiber format to improve buckling resistance of composite plates with central circular holes

    NASA Technical Reports Server (NTRS)

    Hyer, M. W.; Lee, H. H.

    1990-01-01

    The gains in buckling performance are explored that can be achieved by deviating from the conventional straightline fiber format and considering the situation whereby the fiber orientation in a layer, or a group of layers, can vary from point to point. The particular situation studied is a simply supported square plate with a centrally located hole loaded in compression. By using both a sensitivity analysis and a gradient-search technique, fiber orientation in a number of regions of the plate are selected so as to increase the buckling load relative to baseline straightline designs. The sensitivity analysis is used to determine which regions of the plate have the most influence on buckling load, and the gradient search is used to find the design that is believed to represent the absolute maximum buckling load for the conditions prescribed. Convergence studies and sensitivity of the final design are discussed. By examining the stress resultant contours, it is shown how the curvilinear fibers move the load away from the unsupported hole region of the plate to the supported edges, thus increasing the buckling capacity. The tensile capacity of the improved buckling design is investigated, and it is shown that both tensile capacity and buckling capacity can be improved with the curvilinear fiber concept.

  11. [CHARACTERISTICS OF FORMATION, INHIBITION AND DESTRUCTION OF YERSINIA PSEUDOTUBERCULOSIS BIOFILMS FORMING ON ABIOTIC SURFACES].

    PubMed

    Terentieval, N A; Timchenko, N F; Balabanova, L A; Rasskazov, V A

    2015-01-01

    Detection of conditions of Yersinia pseudotuberculosis biofilm formation, their quantitative testing. Y. pseudotuberculosis strains, nutrient media, standard 96-well polystyrene plates, crystal violet dye as well as bacteriologic, spectrophotometric, statistical methods were used. All the studied Y pseudotuberculosis strains formed a well expressed biofilm on abiotic surface during cultivation of bacteria in 200 µl of a plate well at a temperature of 20-22°C for 4-7 days. Bacteria CFU number in biofilm reduced by day 10 of incubation. DNAse I was found to inhibit biofilm formation, and also partially destroyed mature Y. pseudotuberculosis biofilm. The presence of DNA in extra-cellular matrix of biofilm was shown. An ability of Y. pseudotuberculosis to form biofilm on abiotic surface was established. The conditions of biofilm formation were determined. Inhibiting effect of DNAse I on Y. pseudotuberculosis was shown.

  12. 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.

  13. 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.

  14. SNW1 Is a Critical Regulator of Spatial BMP Activity, Neural Plate Border Formation, and Neural Crest Specification in Vertebrate Embryos

    PubMed Central

    Wu, Mary Y.; Ramel, Marie-Christine; Howell, Michael; Hill, Caroline S.

    2011-01-01

    Bone morphogenetic protein (BMP) gradients provide positional information to direct cell fate specification, such as patterning of the vertebrate ectoderm into neural, neural crest, and epidermal tissues, with precise borders segregating these domains. However, little is known about how BMP activity is regulated spatially and temporally during vertebrate development to contribute to embryonic patterning, and more specifically to neural crest formation. Through a large-scale in vivo functional screen in Xenopus for neural crest fate, we identified an essential regulator of BMP activity, SNW1. SNW1 is a nuclear protein known to regulate gene expression. Using antisense morpholinos to deplete SNW1 protein in both Xenopus and zebrafish embryos, we demonstrate that dorsally expressed SNW1 is required for neural crest specification, and this is independent of mesoderm formation and gastrulation morphogenetic movements. By exploiting a combination of immunostaining for phosphorylated Smad1 in Xenopus embryos and a BMP-dependent reporter transgenic zebrafish line, we show that SNW1 regulates a specific domain of BMP activity in the dorsal ectoderm at the neural plate border at post-gastrula stages. We use double in situ hybridizations and immunofluorescence to show how this domain of BMP activity is spatially positioned relative to the neural crest domain and that of SNW1 expression. Further in vivo and in vitro assays using cell culture and tissue explants allow us to conclude that SNW1 acts upstream of the BMP receptors. Finally, we show that the requirement of SNW1 for neural crest specification is through its ability to regulate BMP activity, as we demonstrate that targeted overexpression of BMP to the neural plate border is sufficient to restore neural crest formation in Xenopus SNW1 morphants. We conclude that through its ability to regulate a specific domain of BMP activity in the vertebrate embryo, SNW1 is a critical regulator of neural plate border formation and

  15. Conditional Deletion of Indian Hedgehog in Limb Mesenchyme Results in Complete Loss of Growth Plate Formation but Allows Mature Osteoblast Differentiation.

    PubMed

    Amano, Katsuhiko; Densmore, Michael J; Lanske, Beate

    2015-12-01

    Indian hedgehog (Ihh) is widely recognized as an essential factor for proper skeletal development. Previous in vivo studies using mutant Ihh mouse models were limited by perinatal lethality or carried out after a growth plate formed. Thus the important role of Ihh in mesenchymal cell differentiation has not been investigated. In this study, we established Prx1-Cre;Ihh(fl/fl) mice to ablate Ihh specifically in limb mesenchyme to allow us to observe the phenotype continuously from prenatal development to 3 weeks of age. Mutant mice displayed severe limb abnormalities characterized by complete lack of secondary ossification center and growth plate, indicating an essential role for Ihh in the development of these structures. Interestingly, we discovered that osteoblast differentiation and bone formation could occur in conditions of deficient Ihh. This is a novel finding that has not been observed because of the early lethality of previous Ihh mutants. Mature osteoblasts expressing osteocalcin could be detected in the center of mutant bones at postnatal day 10 (P10). Osteoclasts and blood vessel formation were also present, suggesting active bone remodeling. Histomorphometric analyses show a significant increase in osteoclast number with no major changes in bone formation rate at 3 weeks of age. Mutant long bones in the limbs were deformed, with cortices comprised of irregular woven bone. Also, there was a marked decrease in gene expression of osteoblastic and osteocytic markers. Moreover, mutant long bones displayed bone dysplasia in which we observed increased osteoclast activity and partially reduced osteoblastic and osteocytic differentiation that lead ultimately to loss of bone structures at 3 weeks of age. In summary, our data show for the first time, the presence of mature osteoblasts in long bones of the limbs despite the complete loss of growth plate formation due to Ihh deficiency. These data indicate an important function for Ihh in regulating limb

  16. High-content imaging with micropatterned multiwell plates reveals influence of cell geometry and cytoskeleton on chromatin dynamics.

    PubMed

    Harkness, Ty; McNulty, Jason D; Prestil, Ryan; Seymour, Stephanie K; Klann, Tyler; Murrell, Michael; Ashton, Randolph S; Saha, Krishanu

    2015-10-01

    Understanding the mechanisms underpinning cellular responses to microenvironmental cues requires tight control not only of the complex milieu of soluble signaling factors, extracellular matrix (ECM) connections and cell-cell contacts within cell culture, but also of the biophysics of human cells. Advances in biomaterial fabrication technologies have recently facilitated detailed examination of cellular biophysics and revealed that constraints on cell geometry arising from the cellular microenvironment influence a wide variety of human cell behaviors. Here, we create an in vitro platform capable of precise and independent control of biochemical and biophysical microenvironmental cues by adapting microcontact printing technology into the format of standard six- to 96-well plates to create MicroContact Printed Well Plates (μCP Well Plates). Automated high-content imaging of human cells seeded on μCP Well Plates revealed tight, highly consistent control of single-cell geometry, cytoskeletal organization, and nuclear elongation. Detailed subcellular imaging of the actin cytoskeleton and chromatin within live human fibroblasts on μCP Well Plates was then used to describe a new relationship between cellular geometry and chromatin dynamics. In summary, the μCP Well Plate platform is an enabling high-content screening technology for human cell biology and cellular engineering efforts that seek to identify key biochemical and biophysical cues in the cellular microenvironment.

  17. Chondrocyte-specific knockout of Cbfβ reveals the indispensable function of Cbfβ in chondrocyte maturation, growth plate development and trabecular bone formation in mice.

    PubMed

    Wu, Mengrui; Li, Yi-Ping; Zhu, Guochun; Lu, Yun; Wang, Yiping; Jules, Joel; McConnell, Matthew; Serra, Rosa; Shao, Jian-Zhong; Chen, Wei

    2014-01-01

    Despite years of research into bone formation, the mechanisms by which transcription factors specify growth plate development and trabecular bone formation remain unclear and the role of hypertrophic chondrocytes in trabeculae morphogenesis is controversial. To study the role of Core binding factor beta (Cbfβ) in postnatal cartilage development and endochondral bone formation, we generated chondrocyte-specific Cbfβ-deficient mice (Cbfβf/fCol2α1-Cre mice) using floxed alleles of Cbfβ (Cbfβf/f) and Cre driven by the Collagen 2α1 promoter (Col2α1-Cre). Cbfβf/fCol2α1-Cre mice evaded developmental and newborn lethality to survive to adulthood and displayed severe skeletal malformation. Cbfβf/fCol2α1-Cre mice had dwarfism, hypoplastic skeletons, defective bone mineralization, shortened limbs, shortened sternum bodies, and un-calcified occipital bones and hyoid bones. In the long bone cartilage, the resting zone was elongated, and chondrocyte proliferation and hypertrophy were impaired in Cbfβf/fCol2α1-Cre mice, which led to deformation of the growth plates. Primary spongiosa formation was delayed, diaphysis was shortened and trabecular bone formation was almost absent in the mutant mice. In addition, lamellar bone formation in the secondary spongiosa was also impaired. However, osteoclast formation in the trabecular bone was not affected. Cbfβ deficiency led to down-regulation of chondrocyte-regulating genes [i.e, patched (Ptc1), Cyclin D1 and Indian hedgehog (Ihh)] in the cartilage. Interestingly, the expression of Runx2 and Runx3 was not changed in the cartilage of the mutants. Collectively, the results revealed that Cbfβ is crucial for postnatal skeletal development and endochondral bone formation through its function in growth plate development and chondrocyte proliferation and differentiation. This study also revealed that chondrocyte maturation, mediated by Cbfβ, was critical to trabecular bone morphogenesis. Significantly, these findings provide

  18. Microtiter plate-based antibody microarrays for bacteria and toxins

    USDA-ARS?s Scientific Manuscript database

    Research has focused on the development of rapid biosensor-based, high-throughput, and multiplexed detection of pathogenic bacteria in foods. Specifically, antibody microarrays in 96-well microtiter plates have been generated for the purpose of selective detection of Shiga toxin-producing E. coli (...

  19. Obliquity along plate boundaries

    NASA Astrophysics Data System (ADS)

    Philippon, Mélody; Corti, Giacomo

    2016-12-01

    Most of the plate boundaries are activated obliquely with respect to the direction of far field stresses, as roughly only 8% of the plate boundaries total length shows a very low obliquity (ranging from 0 to 10°, sub-orthogonal to the plate displacement). The obliquity along plate boundaries is controlled by (i) lateral rheological variations within the lithosphere and (ii) consistency with the global plate circuit. Indeed, plate tectonics and magmatism drive rheological changes within the lithosphere and consequently influence strain localization. Geodynamical evolution controls large-scale mantle convection and plate formation, consumption, and re-organization, thus triggering plate kinematics variations, and the adjustment and re-orientation of far field stresses. These geological processes may thus result in plate boundaries that are not perpendicular but oblique to the direction of far field stresses. This paper reviews the global patterns of obliquity along plate boundaries. Using GPlate, we provide a statistical analysis of present-day obliquity along plate boundaries. Within this framework, by comparing natural examples and geological models, we discuss deformation patterns and kinematics recorded along oblique plate boundaries.

  20. Analysis of the effect of transverse power distribution in an involute fuel plate with and without oxide film formation.

    SciTech Connect

    Smith, R. S.

    1998-10-27

    Existing thermal hydraulics computer codes can account for variations in power and temperature in the axial and thickness directions but variations across the width of the plate cannot be accounted for. In the case of fuel plates in an annular core this can lead to significant errors which are accentuated by the presence of an oxide layer that builds up on the aluminum cladding with burnup. This paper uses a three dimensional SINDA model to account for the transverse variations in power. The effect of oxide thickness on these differences is studied in detail. Power distribution and fuel conductivity are also considered. The lower temperatures predicted with the SINDA model result in a greater margin to clad and fuel damage.

  1. 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.

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

    PubMed

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

    2003-07-25

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

  3. Optimization of Salmonella Typhi biofilm assay on polypropylene microtiter plates using response surface methodology.

    PubMed

    Ganjali Dashti, M; Abdeshahian, P; Sudesh, K; Phua, K K

    2016-01-01

    The objective of this study was to develop an optimized assay for Salmonella Typhi biofilm that mimics the environment of the gallbladder as an experimental model for chronic typhoid fever. Multi-factorial assays are difficult to optimize using traditional one-factor-at-a-time optimization methods. Response surface methodology (RSM) was used to optimize six key variables involved in S. Typhi biofilm formation on cholesterol-coated polypropylene 96-well microtiter plates. The results showed that bile (1.22%), glucose (2%), cholesterol (0.05%) and potassium chloride (0.25%) were critical factors affecting the amount of biofilm produced, but agitation (275 rpm) and sodium chloride (0.5%) had antagonistic effects on each other. Under these optimum conditions the maximum OD reading for biofilm formation was 3.4 (λ600 nm), and the coefficients of variation for intra-plate and inter-plate assays were 3% (n = 20) and 5% (n = 8), respectively. These results showed that RSM is an effective approach for biofilm assay optimization.

  4. Formation of {beta}-nickel hydroxide plate-like structures under mild conditions and their optical properties

    SciTech Connect

    Moura, A.P. de; Lima, R.C.; Paris, E.C.; Li, M.S.; Varela, J.A.; Longo, E.

    2011-10-15

    Nanostructural {beta}-nickel hydroxide ({beta}-Ni(OH){sub 2}) plates were prepared using the microwave-hydrothermal (MH) method at a low temperature and short reaction times. An ammonia solution was employed as the coordinating agent, which reacts with [Ni(H{sub 2}O){sub 6}]{sup 2+} to control the growth of {beta}-Ni(OH){sub 2} nuclei. A trigonal {beta}-Ni(OH){sub 2} single phase was observed by X-ray diffraction (XRD) analyses, and the crystal cell was constructed with structural parameters and atomic coordinates obtained from Rietveld refinement. Field emission scanning electron microscopy (FE-SEM) images revealed that the samples consisted of hexagonal-shaped nanoplates with a different particle size distribution. Broad absorption bands assigned as transitions of Ni{sup 2+} in oxygen octahedral sites were revealed by UV-vis spectra. Photoluminescence (PL) properties observed with a maximum peak centered in the blue-green region were attributed to different defects, which were produced during the nucleation process. We present a growth process scheme of the {beta}-Ni(OH){sub 2} nanoplates. - Graphical abstract: Nanostructural {beta}-Ni(OH){sub 2} crystalline powders were prepared by rapid microwave-hydrothermal method for 1, 8 and 32 min. The hexagonal-shaped nanoplates obtained presented PL emission in the blue-green region and each decomposed component represents a different type of electronic transition, which can be linked to the structural arrangement or surface defects. Highlights: > Ammonia solution to control the growth of {beta}-Ni(OH){sub 2} nuclei. > Regular plates-shape related to crystallization-dissolution-recrystallization. > The surface states and lattice defects generated in growth mechanism of crystals. > Different defects produced in the growth process responsible by photoluminescence. > Each component of photoluminescence curve linked to structural arrangement or surface defects.

  5. Plate motion

    SciTech Connect

    Gordon, R.G. )

    1991-01-01

    The motion of tectonic plates on the earth is characterized in a critical review of U.S. research from the period 1987-1990. Topics addressed include the NUVEL-1 global model of current plate motions, diffuse plate boundaries and the oceanic lithosphere, the relation between plate motions and distributed deformations, accelerations and the steadiness of plate motions, the distribution of current Pacific-North America motion across western North America and its margin, plate reconstructions and their uncertainties, hotspots, and plate dynamics. A comprehensive bibliography is provided. 126 refs.

  6. On the actual variety of plate dynamical mechanisms and how mantle evolution affected them through time, from core formation to the Indian collision

    NASA Astrophysics Data System (ADS)

    Osmaston, Miles

    2010-05-01

    If horizontal plate motions were driven by thermal convection of the mantle, they would display the action of slow-to-change body forces. Yet rapid changes of spreading rate and direction, and ridge jumps, are well-documented for the past 130Ma. Also convection cannot readily cause rotations of a plate (e.g. Africa) about a pole within the plate or near it. And plate motions, especially that of India, scarcely fit a convective pattern. To address these problems we look first at mantle motivation at both ends of earth history, beginning with core formation. I then introduce 3 important properties of mantle materials, whose neglect by mantle modellers has surely impaired the value of their work, but whose recognition illuminates the present plate dynamical situation and provides the Earth with a heat engine that is not thermal convection. Finally I sketch the intervening changes in behaviour over time, the sharpest of which brought about the rise of atmospheric oxygen at ~2.25Ga. Core formation. As the very high specific angular momentum of mean planetary material (>105-fold relative to solar) can only be achieved if the planets were wholly accreted in presence of the nebula [1], the iron percolation model is ruled out, because it takes too long. This validates the A.E.Ringwood model (1960-1978) involving nebular H reaction with erupting FeO. The iron then loads the downgoing limb of what is then not a truly thermal convection system. Huge volumes of reaction water were produced, giving the early Earth a wet mantle, a (diminishing) feature that we'll see has constrained mantle behaviour ever since. Plate dynamics since 150Ma. Multiple plate dynamical evidence [2], which will be rapidly re-presented here, shows that currently (a) the Earth has a 2-layer mantle system with a boundary at ~660km and (b) that most cratons have tectospheric keels that reach right to that boundary, or nearly so. The argument is the simple and persuasive one (even to seismologists) of mantle

  7. 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

  8. An Optimized High Throughput Clean-Up Method Using Mixed-Mode SPE Plate for the Analysis of Free Arachidonic Acid in Plasma by LC-MS/MS.

    PubMed

    Wang, Wan; Qin, Suzi; Li, Linsen; Chen, Xiaohua; Wang, Qunjie; Wei, Junfu

    2015-01-01

    A high throughput sample preparation method was developed utilizing mixed-mode solid phase extraction (SPE) in 96-well plate format for the determination of free arachidonic acid in plasma by LC-MS/MS. Plasma was mixed with 3% aqueous ammonia and loaded into each well of 96-well plate. After washing with water and methanol sequentially, 3% of formic acid in acetonitrile was used to elute arachidonic acid. The collected fraction was injected onto a reversed phase column at 30°C with mobile phase of acetonitrile/water (70 : 30, v/v) and detected by LC-MS/MS coupled with electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The calibration curve ranged from 10 to 2500 ng/mL with sufficient linearity (r (2) = 0.9999). The recoveries were in the range of 99.38% to 103.21% with RSD less than 6%. The limit of detection is 3 ng/mL.

  9. D-amino acids do not inhibit Pseudomonas aeruginosa biofilm formation.

    PubMed

    Kao, Wee Tin K; Frye, Mitchell; Gagnon, Patricia; Vogel, Joseph P; Chole, Richard

    2017-02-01

    Pseudomonas aeruginosa, a known biofilm-forming organism, is an opportunistic pathogen that plays an important role in chronic otitis media, tracheitis, cholesteatoma, chronic wounds, and implant infections. Eradication of biofilm infections has been a challenge because the biofilm phenotype provides bacteria with a protective environment from the immune system and antibiotics; thus, there has been great interest in adjunctive molecules that may inhibit biofilm formation or cause biofilm dispersal. There are reports that D-amino acids may inhibit biofilms. In this study, we test the ability of various D-amino acids to inhibit P. aeruginosa biofilm formation in vitro. We evaluated the effect of D-alanine (10 mM), D-leucine (10 mM), D-methionine (10 mM), D-tryptophan (10 mM), and D-tyrosine (10 uM and 1 mM) on biofilm formation in two commonly studied laboratory strains of P. aeruginosa: PAO1 and PA14. Biofilms were grown in 24-well and 96-well tissue culture plates, documented photographically and stained with 0.1% crystal violet and solubilized in 33% glacial acetic acid for quantification. In strains PAO1 and PA14, the addition of D-amino acids did not result in an inhibitory effect on biofilm growth in 24-well plates. Repeating the study in 96-well plates confirmed our findings that D-amino acids do not inhibit biofilm formation of P. aeruginosa. We conclude that D-amino acids only slow the production of biofilms rather than completely prevent biofilm formation; therefore, D-amino acids represent a poor option for potential clinically therapeutic interventions. N/A.

  10. Partial Melt Systems in Plate-Driven Corner Flow: Evaluating the Formation of Porosity Bands as a Mechanism for Magma Focusing at Mid-Ocean Ridges

    NASA Astrophysics Data System (ADS)

    Gebhardt, D.; Butler, S. L.

    2015-12-01

    The imposition of an external shear on a system of partial melt will result in compaction of the solid matrix and concentration of the interstitial liquid melt leading to the formation of regions of contrasting high and low porosity. In experiments, direct and torsional shear geometries have demonstrated that these regions of varying porosity form in bands orientated at low angles relative to the shear plane. A variety of numerical models have been employed to recreate these experimental results. Simple shear, pure shear and torsional shear geometries have been used in both linear and nonlinear numerical settings to model the formation of the porosity bands. In this contribution the numerical models utilize a shear geometry derived from the velocity field of the plate-driven corner flow of a mid-ocean ridge. Motivation for using the velocity field of a mid-ocean ridge comes from evidence that suggests the existence of lateral melt channeling from either side of the ridge axis. Imposing the shear from a mid-ocean ridge corner flow allows for the evaluation of the resulting porosity bands in terms of suitability for channeling melt laterally toward the ridge axis. This is done using both slow and fast spreading ridge geometries. The degree of similarity between previous numerical and experimental results has been found to be greatly influenced by the imposed viscosity law of the solid matrix phase. In order to keep this in mind, the numerical models in this contribution use three different matrix viscosity laws: strain-rate independent, strain-rate dependent and anisotropic. Of these rheologies, strain-rate independence results in the poorest orientation for channeling melt directly to the ridge axis. The strain-rate dependent and anisotropic viscosities present more favorable direct-channeling orientations for the fastest growing porosity bands, but in both cases the background flow will rotate bands to less ideal orientations over time. However, these less

  11. Reduced Staphylococcus aureus biofilm formation in the presence of chitosan-coated iron oxide nanoparticles

    PubMed Central

    Shi, Si-feng; Jia, Jing-fu; Guo, Xiao-kui; Zhao, Ya-ping; Chen, De-sheng; Guo, Yong-yuan; Zhang, Xian-long

    2016-01-01

    Staphylococcus aureus can adhere to most foreign materials and form biofilm on the surface of medical devices. Biofilm infections are difficult to resolve. The goal of this in vitro study was to explore the use of chitosan-coated nanoparticles to prevent biofilm formation. For this purpose, S. aureus was seeded in 96-well plates to incubate with chitosan-coated iron oxide nanoparticles in order to study the efficiency of biofilm formation inhibition. The biofilm bacteria count was determined using the spread plate method; biomass formation was measured using the crystal violet staining method. Confocal laser scanning microscopy and scanning electron microscopy were used to study the biofilm formation. The results showed decreased viable bacteria numbers and biomass formation when incubated with chitosan-coated iron oxide nanoparticles at all test concentrations. Confocal laser scanning microscopy showed increased dead bacteria and thinner biofilm when incubated with nanoparticles at a concentration of 500 µg/mL. Scanning electron microscopy revealed that chitosan-coated iron oxide nanoparticles inhibited biofilm formation in polystyrene plates. Future studies should be performed to study these nanoparticles for anti-infective use. PMID:27994455

  12. Reduced Staphylococcus aureus biofilm formation in the presence of chitosan-coated iron oxide nanoparticles.

    PubMed

    Shi, Si-Feng; Jia, Jing-Fu; Guo, Xiao-Kui; Zhao, Ya-Ping; Chen, De-Sheng; Guo, Yong-Yuan; Zhang, Xian-Long

    Staphylococcus aureus can adhere to most foreign materials and form biofilm on the surface of medical devices. Biofilm infections are difficult to resolve. The goal of this in vitro study was to explore the use of chitosan-coated nanoparticles to prevent biofilm formation. For this purpose, S. aureus was seeded in 96-well plates to incubate with chitosan-coated iron oxide nanoparticles in order to study the efficiency of biofilm formation inhibition. The biofilm bacteria count was determined using the spread plate method; biomass formation was measured using the crystal violet staining method. Confocal laser scanning microscopy and scanning electron microscopy were used to study the biofilm formation. The results showed decreased viable bacteria numbers and biomass formation when incubated with chitosan-coated iron oxide nanoparticles at all test concentrations. Confocal laser scanning microscopy showed increased dead bacteria and thinner biofilm when incubated with nanoparticles at a concentration of 500 µg/mL. Scanning electron microscopy revealed that chitosan-coated iron oxide nanoparticles inhibited biofilm formation in polystyrene plates. Future studies should be performed to study these nanoparticles for anti-infective use.

  13. The formation of a hybrid structure from tungsten selenide and oxide plates for a hydrogen-evolution electrocatalyst

    NASA Astrophysics Data System (ADS)

    Fominski, V. Yu.; Grigoriev, S. N.; Romanov, R. I.; Volosova, M. A.; Grunin, A. I.; Teterina, G. D.

    2016-06-01

    It has been found that the pulsed laser deposition of a thin tungsten selenide film, followed by thermal treatment at 550°C in an Ar + O2 mixture of gases, results in the formation of a hybrid structure that is made up of ultrathin WSe2 and WO3- y platelets. The structural and size characteristics of the nanoplatelets deposited on microcrystalline graphite provide the effective hydrogen evolution reaction in a 0.5 M H2SO4 solution, with the cathode current made about seven times higher at a potential of-100 mV and the slope of the Tafel characteristic reduced from 340 to 90 mV/dec.

  14. Comparison study of streptavidin-coated microtitration plates.

    PubMed

    Välimaa, Lasse; Laurikainen, Katja

    2006-01-20

    Streptavidin (SAv)-coated 96-well microtitration plates from commercial suppliers (six plates) and our laboratory (two plates) were tested with respect to their binding capacity for small and large molecules, leaching, well-to-well variation and immunoassay performance. The binding capacities for europium-labelled biotin (Eu-biotin) varied from 4.4 to above 150 pmol/well depending on the plate, and for biotinylated monoclonal antibody from 1.2 to 6.4 pmol/well (190-1030 ng). Incubation for 1 h in regular immunoassay buffer resulted in leaching of 0.6-76 ng of SAv from unwashed wells and 0.5-60 ng from pre-washed wells. The desorbed quantity represented up to 3.4% of the maximal Eu-biotin binding capacity. Coating-related variation of 96 wells (CV% values) ranged from 1.2% to 8.0% when tested with respect to maximum binding capacity. Immunofluorometric assay for TSH as well as enzyme immunoassays for CA125 and PSA revealed immunoassay-related performance of the plates, regarding signal levels, variation and non-specific binding characteristics. Non-specific binding and variation tended to increase in the highest capacity plates, whereas some low capacity plates fulfilled the assay requirements optimally. Good performance with respect to capacity, coating homogeneity or leaching was not by definition reflected in the performance of two-site heterogeneous immunoassays.

  15. 49 CFR 451.23 - Plate specifications.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 6 2013-10-01 2013-10-01 false Plate specifications. 451.23 Section 451.23... SECURITY SAFETY APPROVAL OF CARGO CONTAINERS TESTING AND APPROVAL OF CONTAINERS Safety Approval Plate § 451.23 Plate specifications. (a) The safety approval plate must be of the size and in the format...

  16. 49 CFR 451.23 - Plate specifications.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 6 2014-10-01 2014-10-01 false Plate specifications. 451.23 Section 451.23... SECURITY SAFETY APPROVAL OF CARGO CONTAINERS TESTING AND APPROVAL OF CONTAINERS Safety Approval Plate § 451.23 Plate specifications. (a) The safety approval plate must be of the size and in the format...

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

    PubMed

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

    2009-08-15

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

  18. Formation and metasomatism of continental lithospheric mantle in intra-plate and subduction-related tectonic settings

    NASA Astrophysics Data System (ADS)

    Ionov, Dmitri

    2010-05-01

    , major and trace element and isotope compositions of fertile lherzolites and thus cannot provide viable alternatives to the concept of melt extraction from pristine mantle as the major mechanism of CLM formation. Published data on xenoliths from andesitic volcanoes and on supra-subduction oceanic peridotites [4] show that the most common rocks in mantle wedge lithosphere are highly refractory harzburgites characterized by a combination of variable but generally high modal opx (18-30%) with very low modal cpx (1.5-3%). At a given olivine (or MgO) content, they have higher opx and silica, and lower cpx, Al and Ca contents than normal refractory peridotite xenoliths in continental basalts; the Mg-Si and Al-Si trends in those rocks resemble those in cratonic peridotites. These features may indicate either fluid fluxing during melting in the mantle wedge or selective post-melting metasomatic enrichments in silica to transform some olivine to opx. High oxygen fugacities and radiogenic Os-isotope compositions in those rocks may be related to enrichments by slab-derived fluids, but these features are not always coupled with trace element enrichments or patterns commonly attributed to "subduction zone metasomatism" deduced from studies of arc volcanic rocks and experiments. The valuable insights provided by experimental work and xenolith case studies are difficult to apply to many natural peridotite series because late-stage processes commonly overlap the evidence for initial melting. References: [1] Herzberg C., J. Petrol. 45: 2507 (2004). [2] Ionov D. & Sobolev A., GCA 72 (S1): A410 (2008). [3] Ionov D., Contrib. Miner. Petrol. (2007) [4] Ionov D., J. Petrol. doi: 10.1093/petrology/egp090 (2010)

  19. Microtiter plate transformation using the LiAc/SS carrier DNA/PEG method.

    PubMed

    Gietz, R Daniel; Schiestl, Robert H

    2007-01-01

    Here, we describe a protocol that has been adapted for the transformation of yeast cells in 96-well microtiter plates. This protocol can be tailored for multiple applications and is suitable for high-throughput applications. It can be completed in 2-3 h, once the yeast cells have been grown depending on the heat shock used.

  20. Broken formations, melanges and olistostromes in Puerto Plata area (Northern Dominican Republic) as a record of subduction and collisional processes between the Caribbean and North-American plates

    NASA Astrophysics Data System (ADS)

    Hernaiz Huerta, Pedro Pablo; Valera Fernando, Pérez; de Los Santos Manuel, Abad; Jacques, Monthel; de Neira Alberto, Díaz

    2010-05-01

    The Northern Cordillera of the Dominican Republic records the (oblique) subduction and collisional processes occurred between the Caribbean and North-American plates during Upper Cretaceous to Lower Paleogene times. The boundary between these two plates can be traced within this range disrupted by an Upper Paleogene to present intense left-lateral strike-slip tectonism, onset after collision. In the western part of the range this boundary might be defined by the Camu fault. In the coastal area of Puerto Plata, located on the northern block of the Camu fault, basement rocks belonging to the subducting plate (the Puerto Plata Basement Complex) and several related units probably formed in an accretionary prism, preserve in a large (300 km2) outcrop of chaotic formations presumably formed (and exhumed) during collision. They include from broken formations and tectonic melanges to olistostromes and other coeval sedimentary deposits. The Puerto Plata Basement Complex (PPBC) consists of highly faulted and dismembered blocks formed by discontinuous but sometimes coherent outcrops of serpentinized or massive peridotite, pods of ultramafic cumulates, massive or banded gabbros and Los Caños Fm, a thick sequence of gross bedded volcaniclastic material with interbedded basaltic (sometimes pillowed) or andesitic flows. All these rocks bear low grade metamorphism and lack a general deformation fabric apart from occasional transformation to mylonites due to localized shearing. The PPBC has been interpreted as a fragment of oceanic crust, belonging to the subducting (North-American) slab that has been exhumed as a tectonic melange or a broken complex. The Imbert Fm, of Palaeocene-Eocene age, is formed by a well bedded succession of white very fine grained porcelaneous tuffs, with eventual intercalations of cherts, limestones and marls that, towards the lower part, is interbedded with volcanic-derived graywackes and limolites, and more occasionally, thick beds of conglomerates and

  1. Long-term Formation of Aggressive Bony Lesions in Dogs with Mid-Diaphyseal Fractures Stabilized with Metallic Plates: Incidence in a Tertiary Referral Hospital Population

    PubMed Central

    Gilley, Robert S.; Hiebert, Elizabeth; Clapp, Kemba; Bartl-Wilson, Lara; Nappier, Michael; Werre, Stephen; Barnes, Katherine

    2017-01-01

    The incidence of complications secondary to fracture stabilization, particularly osteolytic lesions and bony tumor formation, has long been difficult to evaluate. The objective of this study was to describe the long-term incidence of aggressive bony changes developing in dogs with long bone diaphyseal fractures stabilized by metallic bone plates compared to a breed-, sex-, and age-matched control group. The medical records of a tertiary referral center were retrospectively reviewed for dogs that matched each respective criterion. Signalment, history, cause of death (if applicable), and aggressive bony changes at previous fracture sites were recorded. Ninety dogs met the criteria for inclusion in the fracture group and were matched with appropriate control dogs. Four of the dogs in the fracture group developed aggressive bony changes at the site of previous fracture repairs most consistent with osseous neoplasia. One lesion was confirmed with cytology as neoplastic. The population of dogs was mixed with regard to breed and body weight, but all dogs with aggressive bony lesions were male. Incidence of aggressive bony lesion formation in the fracture group was 4 (4.4%) and was 0 (0%) in the control group; three (75%) of the affected dogs in the fracture group included cerclage as a component of their primary fracture stabilizations. Incidence of aggressive bony lesions in the fracture group compared to the control group was determined to be statistically significant (p = 0.0455), as was the incidence of cerclage among dogs affected by aggressive bony lesions compared to the rest of the fracture group (p = 0.0499). Development of aggressive bony lesions is an uncommon complication of fracture fixation. Additional research is needed to further identify and elucidate the long-term effects of metallic implants in dogs. PMID:28197406

  2. Long-term Formation of Aggressive Bony Lesions in Dogs with Mid-Diaphyseal Fractures Stabilized with Metallic Plates: Incidence in a Tertiary Referral Hospital Population.

    PubMed

    Gilley, Robert S; Hiebert, Elizabeth; Clapp, Kemba; Bartl-Wilson, Lara; Nappier, Michael; Werre, Stephen; Barnes, Katherine

    2017-01-01

    The incidence of complications secondary to fracture stabilization, particularly osteolytic lesions and bony tumor formation, has long been difficult to evaluate. The objective of this study was to describe the long-term incidence of aggressive bony changes developing in dogs with long bone diaphyseal fractures stabilized by metallic bone plates compared to a breed-, sex-, and age-matched control group. The medical records of a tertiary referral center were retrospectively reviewed for dogs that matched each respective criterion. Signalment, history, cause of death (if applicable), and aggressive bony changes at previous fracture sites were recorded. Ninety dogs met the criteria for inclusion in the fracture group and were matched with appropriate control dogs. Four of the dogs in the fracture group developed aggressive bony changes at the site of previous fracture repairs most consistent with osseous neoplasia. One lesion was confirmed with cytology as neoplastic. The population of dogs was mixed with regard to breed and body weight, but all dogs with aggressive bony lesions were male. Incidence of aggressive bony lesion formation in the fracture group was 4 (4.4%) and was 0 (0%) in the control group; three (75%) of the affected dogs in the fracture group included cerclage as a component of their primary fracture stabilizations. Incidence of aggressive bony lesions in the fracture group compared to the control group was determined to be statistically significant (p = 0.0455), as was the incidence of cerclage among dogs affected by aggressive bony lesions compared to the rest of the fracture group (p = 0.0499). Development of aggressive bony lesions is an uncommon complication of fracture fixation. Additional research is needed to further identify and elucidate the long-term effects of metallic implants in dogs.

  3. Prevention of Staphylococcus aureus biofilm formation by antibiotics in 96-Microtiter Well Plates and Drip Flow Reactors: critical factors influencing outcomes

    PubMed Central

    Manner, Suvi; Goeres, Darla M.; Skogman, Malena; Vuorela, Pia; Fallarero, Adyary

    2017-01-01

    Biofilm formation leads to the failure of antimicrobial therapy. Thus, biofilm prevention is a desirable goal of antimicrobial research. In this study, the efficacy of antibiotics (doxycycline, oxacillin and rifampicin) in preventing Staphylococcus aureus biofilms was investigated using Microtiter Well Plates (MWP) and Drip Flow Reactors (DFR), two models characterized by the absence and the presence of a continuous flow of nutrients, respectively. Planktonic culture of S. aureus was exposed to antibiotics for one hour followed by 24 hours incubation with fresh nutrients in MWP or continuous flow of nutrients in DFR. The DFR grown biofilms were significantly more tolerant to the antibiotics than those grown in MWP without the continuous flow. The differences in log reductions (LR) between the two models could not be attributed to differences in the cell density, the planktonic inoculum concentration or the surface-area-to-volume ratios. However, eliminating the flow in the DFR significantly restored the antibiotic susceptibility. These findings demonstrate the importance of considering differences between experimental conditions in different model systems, particularly the flow of nutrients, when performing anti-biofilm efficacy evaluations. Biofilm antibiotic efficacy studies should be assessed using various models and more importantly, in a model mimicking conditions of its clinical application. PMID:28252025

  4. Establishment of a miniaturized enzyme-linked immunosorbent assay for human transferrin quantification using an intelligent multifunctional analytical plate.

    PubMed

    Spies, Peter; Chen, Guo Jun; Gygax, Daniel

    2008-11-01

    A miniaturized enzyme-linked immunosorbent assay (ELISA) with a reaction volume of 5 microl for human transferrin quantification has successfully been developed using an intelligent multifunctional analytical plate (IMAPlate 5RC96), the first miniature analytical platform capable of manually performing parallel liquid transfer, reaction, and analysis. This is the first article to validate the platform for the ELISA application. The data obtained from the standards in this miniaturized ELISA can well be fitted by a one-site binding reaction mode, the coefficient of variation (CV) of the whole plate for an artificial sample (spiking a known concentration of human transferrin into the assay diluent) is 7.0%, and the mean recovery is between 94 and 114% (n=96), comparable to the values from conventional ELISA in a 96-well format plate. The IMAPlate 5RC96-based miniaturized ELISA not only can reduce sample and reagent consumption to 5% of the conventional ELISA but also can shorten the reaction time. Combined with the advantages brought by miniaturization, the easy-to-handle, parallel, and simultaneous liquid transfer features of the IMAPlate 5RC96 provide a completely new lab tool for manually performing high-throughput ELISA. Our results demonstrate that the IMAPlate 5RC96 is a convenient, robust, high-throughput lab device feasible for miniaturized ELISA in an ordinary laboratory.

  5. A kinematic model for the formation of the Siletz-Crescent forearc terrane by capture of coherent fragments of the Farallon and Resurrection plates

    NASA Astrophysics Data System (ADS)

    McCrory, Patricia A.; Wilson, Douglas S.

    2013-06-01

    volcanic basement of the Oregon and Washington Coast ranges has been proposed to represent a pair of tracks of the Yellowstone hotspot formed at a mid-ocean ridge during the early Cenozoic. This interpretation has been questioned on many grounds, especially that the range of ages does not match the offshore spreading rates and that the presence of continental coarse clastic sediments is difficult to reconcile with fast convergence rates between the oceanic plates and North America. Updates to basement geochronology and plate motion history reveal that these objections are much less serious than when they were first raised. Forward plate kinematic modeling reveals that predicted basement ages can be consistent with the observed range of about 55-49 Ma, and that the entire basement terrane can form within about 300 km of continental sources for clastic sediments. This kinematic model indicates that there is no firm reason to reject the near-ridge hotspot hypothesis on the basis of plate motions. A novel element of the model is the Resurrection plate, previously proposed to exist between the Farallon and Kula plates. By including the defunct Resurrection plate in our reconstruction, we are able to model the Farallon hotspot track as docking against the Oregon subduction margin starting about 53 Ma, followed by docking of the Resurrection track to the north starting about 48 Ma. Accretion of the Farallon plate fragment and partial subduction of the Resurrection fragment complicates the three-dimensional structure of the modern Cascadia forearc. We interpret the so-called "E" layer beneath Vancouver Island to be part of the Resurrection fragment. Our new kinematic model of mobile terranes within the Paleogene North American plate boundary allows reinterpretation of the three-dimensional structure of the Cascadia forearc and its relationship to ongoing seismotectonic processes.

  6. A kinematic model for the formation of the Siletz-Crescent forearc terrane by capture of coherent fragments of the Farallon and Resurrection plates

    USGS Publications Warehouse

    McCrory, Patricia A.; Wilson, Douglas S.

    2013-01-01

    The volcanic basement of the Oregon and Washington Coast ranges has been proposed to represent a pair of tracks of the Yellowstone hotspot formed at a mid-ocean ridge during the early Cenozoic. This interpretation has been questioned on many grounds, especially that the range of ages does not match the offshore spreading rates and that the presence of continental coarse clastic sediments is difficult to reconcile with fast convergence rates between the oceanic plates and North America. Updates to basement geochronology and plate motion history reveal that these objections are much less serious than when they were first raised. Forward plate kinematic modeling reveals that predicted basement ages can be consistent with the observed range of about 55–49 Ma, and that the entire basement terrane can form within about 300 km of continental sources for clastic sediments. This kinematic model indicates that there is no firm reason to reject the near-ridge hotspot hypothesis on the basis of plate motions. A novel element of the model is the Resurrection plate, previously proposed to exist between the Farallon and Kula plates. By including the defunct Resurrection plate in our reconstruction, we are able to model the Farallon hotspot track as docking against the Oregon subduction margin starting about 53 Ma, followed by docking of the Resurrection track to the north starting about 48 Ma. Accretion of the Farallon plate fragment and partial subduction of the Resurrection fragment complicates the three-dimensional structure of the modern Cascadia forearc. We interpret the so-called “E” layer beneath Vancouver Island to be part of the Resurrection fragment. Our new kinematic model of mobile terranes within the Paleogene North American plate boundary allows reinterpretation of the three-dimensional structure of the Cascadia forearc and its relationship to ongoing seismotectonic processes.

  7. Peen plating

    NASA Technical Reports Server (NTRS)

    Babecki, A. J. (Inventor); Haehner, C. L.

    1973-01-01

    A process for metal plating which comprises spraying a mixture of metallic powder and small peening particles at high velocity against a surface is described. The velocity must be sufficient to impact and bond metallic powder onto the surface. In the case of metal surfaces, the process has as one of its advantages providing mechanical working (hardening) of the surface simultaneously with the metal plating.

  8. Evidence for Sexual Dimorphism in the Plated Dinosaur Stegosaurus mjosi (Ornithischia, Stegosauria) from the Morrison Formation (Upper Jurassic) of Western USA.

    PubMed

    Saitta, Evan Thomas

    2015-01-01

    Conclusive evidence for sexual dimorphism in non-avian dinosaurs has been elusive. Here it is shown that dimorphism in the shape of the dermal plates of Stegosaurus mjosi (Upper Jurassic, western USA) does not result from non-sex-related individual, interspecific, or ontogenetic variation and is most likely a sexually dimorphic feature. One morph possessed wide, oval plates 45% larger in surface area than the tall, narrow plates of the other morph. Intermediate morphologies are lacking as principal component analysis supports marked size- and shape-based dimorphism. In contrast, many non-sex-related individual variations are expected to show intermediate morphologies. Taphonomy of a new quarry in Montana (JRDI 5ES Quarry) shows that at least five individuals were buried in a single horizon and were not brought together by water or scavenger transportation. This new site demonstrates co-existence, and possibly suggests sociality, between two morphs that only show dimorphism in their plates. Without evidence for niche partitioning, it is unlikely that the two morphs represent different species. Histology of the new specimens in combination with studies on previous specimens indicates that both morphs occur in fully-grown individuals. Therefore, the dimorphism is not a result of ontogenetic change. Furthermore, the two morphs of plates do not simply come from different positions on the back of a single individual. Plates from all positions on the body can be classified as one of the two morphs, and previously discovered, isolated specimens possess only one morph of plates. Based on the seemingly display-oriented morphology of plates, female mate choice was likely the driving evolutionary mechanism rather than male-male competition. Dinosaur ornamentation possibly served similar functions to the ornamentation of modern species. Comparisons to ornamentation involved in sexual selection of extant species, such as the horns of bovids, may be appropriate in predicting the

  9. Evidence for Sexual Dimorphism in the Plated Dinosaur Stegosaurus mjosi (Ornithischia, Stegosauria) from the Morrison Formation (Upper Jurassic) of Western USA

    PubMed Central

    Saitta, Evan Thomas

    2015-01-01

    Conclusive evidence for sexual dimorphism in non-avian dinosaurs has been elusive. Here it is shown that dimorphism in the shape of the dermal plates of Stegosaurus mjosi (Upper Jurassic, western USA) does not result from non-sex-related individual, interspecific, or ontogenetic variation and is most likely a sexually dimorphic feature. One morph possessed wide, oval plates 45% larger in surface area than the tall, narrow plates of the other morph. Intermediate morphologies are lacking as principal component analysis supports marked size- and shape-based dimorphism. In contrast, many non-sex-related individual variations are expected to show intermediate morphologies. Taphonomy of a new quarry in Montana (JRDI 5ES Quarry) shows that at least five individuals were buried in a single horizon and were not brought together by water or scavenger transportation. This new site demonstrates co-existence, and possibly suggests sociality, between two morphs that only show dimorphism in their plates. Without evidence for niche partitioning, it is unlikely that the two morphs represent different species. Histology of the new specimens in combination with studies on previous specimens indicates that both morphs occur in fully-grown individuals. Therefore, the dimorphism is not a result of ontogenetic change. Furthermore, the two morphs of plates do not simply come from different positions on the back of a single individual. Plates from all positions on the body can be classified as one of the two morphs, and previously discovered, isolated specimens possess only one morph of plates. Based on the seemingly display-oriented morphology of plates, female mate choice was likely the driving evolutionary mechanism rather than male-male competition. Dinosaur ornamentation possibly served similar functions to the ornamentation of modern species. Comparisons to ornamentation involved in sexual selection of extant species, such as the horns of bovids, may be appropriate in predicting the

  10. Imaging and quantitative analysis of tritium-labelled cells in lymphocyte proliferation assays using microchannel plate detectors originally developed for X-ray astronomy.

    PubMed

    Lees, J E; Hales, J M

    2001-01-01

    Microchannel plate detectors have been used in many astronomical X-ray telescopes. Recently we have begun to use similar detectors to image electron emission from radiolabelled biological assays. Here we show how a microchannel plate (MCP) detector can be used to image tritium uptake in T lymphocyte proliferation assays. Quantitative analysis using the MCP detector has the same sensitivity and speed as conventional liquid scintillation counter (LSC) analysis whilst obviating the need for scintillation fluid. In addition the system permits the imaging of whole plate harvests from a range of plate sizes. Here we present data obtained with 96-well plates and Terasaki plates.

  11. Tectonics of the Easter plate

    NASA Technical Reports Server (NTRS)

    Engeln, J. F.; Stein, S.

    1984-01-01

    A new model for the Easter plate is presented in which rift propagation has resulted in the formation of a rigid plate between the propagating and dying ridges. The distribution of earthquakes, eleven new focal mechanisms, and existing bathymetric and magnetic data are used to describe the tectonics of this area. Both the Easter-Nazca and Easter-Pacific Euler poles are sufficiently close to the Easter plate to cause rapid changes in rates and directions of motion along the boundaries. The east and west boundaries are propagating and dying ridges; the southwest boundary is a slow-spreading ridge and the northern boundary is a complex zone of convergent and transform motion. The Easter plate may reflect the tectonics of rift propagation on a large scale, where rigid plate tectonics requires boundary reorientation. Simple schematic models to illustrate the general features and processes which occur at plates resulting from large-scale rift propagation are used.

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

    PubMed Central

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

    2015-01-01

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

  13. A cost-effective plate-based sample preparation for antibody N-glycan analysis.

    PubMed

    Burnina, Irina; Hoyt, Erik; Lynaugh, Heather; Li, Huijuan; Gong, Bing

    2013-09-13

    During early cell line and process development of therapeutic antibodies, a cost-effective high-throughput approach to characterize the N-linked glycans is highly desired given that a large number of samples need to be analyzed. Using commercially available, low cost 96-well plates, we developed a practical procedure to prepare fluorescently labeled N-linked glycans for both qualitative and quantitative analysis by mass spectrometry (MS) and ultrahigh performance liquid chromatography (UPLC). Antibody samples were continuously denatured, reduced, and deglycosylated in a single 96-well hydrophobic membrane filter plate. Subsequently, released glycans were fluorescently labeled in a collection plate, and cleaned-up using a hydrophilic membrane filter plate. Carried out entirely in ready-to-use 96-well plates with simple buffer systems, this procedure requires less than 90min to finish. We applied the optimized procedure to examine the N-linked glycosylation of trastuzumab and were able to quantify ten major N-linked glycans. The results from different amounts of starting materials (10-200μg) were highly similar and showed the robustness of this procedure. Compared to other methods, this new procedure is simple to implement, economically more affordable, and could be very valuable for early screenings of antibody development.

  14. Nitrided Metallic Bipolar Plates

    SciTech Connect

    Brady, Michael P; Tortorelli, Peter F; Pihl, Josh A; Toops, Todd J; More, Karren Leslie; Meyer III, Harry M; Vitek, John Michael; Wang, Heli; Turner, John; Wilson, Mahlon; Garzon, Fernando; Rockward, Tommy; Connors, Dan; Rakowski, Jim; Gervasio, Don

    2008-01-01

    The objectives are: (1) Develop and optimize stainless steel alloys amenable to formation of a protective Cr-nitride surface by gas nitridation, at a sufficiently low cost to meet DOE targets and with sufficient ductility to permit manufacture by stamping. (2) Demonstrate capability of nitridation to yield high-quality stainless steel bipolar plates from thin stamped alloy foils (no significant stamped foil warping or embrittlement). (3) Demonstrate single-cell fuel cell performance of stamped and nitrided alloy foils equivalent to that of machined graphite plates of the same flow-field design ({approx}750-1,000 h, cyclic conditions, to include quantification of metal ion contamination of the membrane electrode assembly [MEA] and contact resistance increase attributable to the bipolar plates). (4) Demonstrate potential for adoption in automotive fuel cell stacks. Thin stamped metallic bipolar plates offer the potential for (1) significantly lower cost than currently-used machined graphite bipolar plates, (2) reduced weight/volume, and (3) better performance and amenability to high volume manufacture than developmental polymer/carbon fiber and graphite composite bipolar plates. However, most metals exhibit inadequate corrosion resistance in proton exchange membrane fuel cell (PEMFC) environments. This behavior leads to high electrical resistance due to the formation of surface oxides and/or contamination of the MEA by metallic ions, both of which can significantly degrade fuel cell performance. Metal nitrides offer electrical conductivities up to an order of magnitude greater than that of graphite and are highly corrosion resistant. Unfortunately, most conventional coating methods (for metal nitrides) are too expensive for PEMFC stack commercialization or tend to leave pinhole defects, which result in accelerated local corrosion and unacceptable performance.

  15. Importance of the Small-Scale Processes Melting, Plate Boundary Formation and Mineralogy on the Large-Scale, Long-Term Thermo-Chemical Evolution of Earth's Mantle-Plate System

    NASA Astrophysics Data System (ADS)

    Tackley, P.

    2015-12-01

    Seismic observations of the deep Earth reveal the presence of two large low shear velocity provinces (LLSVPs) that are typically inferred to be dense chemically-distinct material, as well as discontinuities that are typically linked to the post-perovskite (pPv) phase transition. Several possible origins of chemically-dense material have been proposed, including recycling of mid-ocean ridge basalt (MORB), primordial differentiation events, crystallisation of a basal magma ocean, or some combination of these creating a basal melange (BAM; Tackley 2012 Earth Sci. Rev.). Each of these possibilities would result in a different composition hence different mineralogy. In order to constrain this we have been running calculations of thermo-chemical mantle evolution over 4.5 billion years that include melting-induced differentiation, plate tectonics induced by strongly temperature-dependent viscosity and plastic yielding, core cooling and compressibility with reasonable assumptions about the pressure-dependence of other material properties. Some of our simulations start from a magma ocean state so initial layering is developed self-consistently. Already-published results (Nakagawa et al., 2009 GCubed, 2010 PEPI, 2012 GCubed) already indicate the importance of exact MORB composition on the amount of MORB segregating above the CMB, which in turn influences mantle thermal structure and the evolution of the core and geodynamo. In more recent results we have been additionally including primordial material. We find that melting-induced differentiation has several first-order effects on the dynamics, including (i) making plate tectonics easier (through stresses associated with lateral variations in crustal thickness) and (ii) reducing heat flux through the CMB (due to the build-up of dense material above the CMB); also (iii) tectonic mode (continuous plate tectonics, episodic lid or stagnant lid) also makes a first-order difference to mantle structure and dynamics. This emphasises

  16. Robotic platform for parallelized cultivation and monitoring of microbial growth parameters in microwell plates.

    PubMed

    Knepper, Andreas; Heiser, Michael; Glauche, Florian; Neubauer, Peter

    2014-12-01

    The enormous variation possibilities of bioprocesses challenge process development to fix a commercial process with respect to costs and time. Although some cultivation systems and some devices for unit operations combine the latest technology on miniaturization, parallelization, and sensing, the degree of automation in upstream and downstream bioprocess development is still limited to single steps. We aim to face this challenge by an interdisciplinary approach to significantly shorten development times and costs. As a first step, we scaled down analytical assays to the microliter scale and created automated procedures for starting the cultivation and monitoring the optical density (OD), pH, concentrations of glucose and acetate in the culture medium, and product formation in fed-batch cultures in the 96-well format. Then, the separate measurements of pH, OD, and concentrations of acetate and glucose were combined to one method. This method enables automated process monitoring at dedicated intervals (e.g., also during the night). By this approach, we managed to increase the information content of cultivations in 96-microwell plates, thus turning them into a suitable tool for high-throughput bioprocess development. Here, we present the flowcharts as well as cultivation data of our automation approach.

  17. Development of Multiwell-Plate Methods Using Pure Cultures of Methanogens To Identify New Inhibitors for Suppressing Ruminant Methane Emissions.

    PubMed

    Weimar, M R; Cheung, J; Dey, D; McSweeney, C; Morrison, M; Kobayashi, Y; Whitman, W B; Carbone, V; Schofield, L R; Ronimus, R S; Cook, G M

    2017-08-01

    Hydrogenotrophic methanogens typically require strictly anaerobic culturing conditions in glass tubes with overpressures of H2 and CO2 that are both time-consuming and costly. To increase the throughput for screening chemical compound libraries, 96-well microtiter plate methods for the growth of a marine (environmental) methanogen Methanococcus maripaludis strain S2 and the rumen methanogen Methanobrevibacter species AbM4 were developed. A number of key parameters (inoculum size, reducing agents for medium preparation, assay duration, inhibitor solvents, and culture volume) were optimized to achieve robust and reproducible growth in a high-throughput microtiter plate format. The method was validated using published methanogen inhibitors and statistically assessed for sensitivity and reproducibility. The Sigma-Aldrich LOPAC library containing 1,280 pharmacologically active compounds and an in-house natural product library (120 compounds) were screened against M. maripaludis as a proof of utility. This screen identified a number of bioactive compounds, and MIC values were confirmed for some of them against M. maripaludis and M. AbM4. The developed method provides a significant increase in throughput for screening compound libraries and can now be used to screen larger compound libraries to discover novel methanogen-specific inhibitors for the mitigation of ruminant methane emissions.IMPORTANCE Methane emissions from ruminants are a significant contributor to global greenhouse gas emissions, and new technologies are required to control emissions in the agriculture technology (agritech) sector. The discovery of small-molecule inhibitors of methanogens using high-throughput phenotypic (growth) screening against compound libraries (synthetic and natural products) is an attractive avenue. However, phenotypic inhibitor screening is currently hindered by our inability to grow methanogens in a high-throughput format. We have developed, optimized, and validated a high

  18. Improved microbioassay for plasma erythropoietin based on CFU-E colony formation.

    PubMed

    Sakata, S; Enoki, Y

    1992-05-01

    We examined the conditions necessary for performing a reliable erythropoietin (EPO) assay based on CFU-E colony formation in fetal mouse liver cell (FMLC) microcultures using 96-well microtiter plates. Both linearity of colony numbers with the number of cells plated and comparison among the colony ratios at various densities of seeding cells indicated that the colonies originated from a single progenitor cell when 7500 or fewer cells were plated into individual microtiter wells. About a twofold CFU-E enrichment in 12- to 13-day FMLC was achieved by Ficoll-Paque centrifugation. Plasma treated with acid-boiling stimulated the colony formation most and contained no colony inhibitor. Dose-response curve for the plasma was parallel to the EPO standard curve. The "erythroid colony-stimulating activity" in the plasma was additive to that in the standard EPO, and was completely neutralized by a monoclonal antibody against recombinant human EPO. Using the assay procedure thus established, plasma EPO titer was determined in normal subjects, in patients with nonuremic anemia and polycythemia vera, and in dialysis patients with chronic renal failure. The use of different preparations of standard EPO resulted in a significant difference in the titers because their dose-response curves differed from one another. An inverse relationship was found between EPO titers and hemoglobin concentrations in the nonuremic anemic patients, but not in the dialysis patients with about one half the normal EPO level.

  19. Comparison of methods for the detection of biofilm formation by Staphylococcus aureus isolated from bovine subclinical mastitis

    PubMed Central

    de Castro Melo, Poliana; Ferreira, Luciano Menezes; Filho, Antônio Nader; Zafalon, Luiz Francisco; Vicente, Hinig Isa Godoy; de Souza, Viviane

    2013-01-01

    Biofilm formation is considered to be a selective advantage for Staphylococcus aureus mastitis isolates by facilitating bacterial persistence in the udder. It requires attachment to mammary epithelium, proliferation and accumulation of cells in multilayers. The objective of this study was to determine the sensitivity and specificity of three techniques for the detection of S. aureus biofilm-positive strains. Two phenotypic tests, including growth on microtitre plates and Congo red agar, were compared with a PCR technique using 94 S. aureus strains obtained from cows with subclinical mastitis from two farms in the state of São Paulo. These strains were characterised by in vitro slime production on Congo red agar, biofilm formation on microtitre plates and the presence of the icaA and icaD genes. The results revealed that 85% of the isolates tested produced slime on the Congo red agar, 98.9% of the isolates produced biofilms in vitro by adhering to sterile 96-well “U” bottom polystyrene tissue culture plates, and 95.7% of the isolates carried the icaA and icaD genes. The results of the phenotypic tests for biofilm formation were compared with those of the molecular analysis, and the sensitivity and specificity of the Congo red agar test were 88.9% and 100%, respectively, while those of the microtitre plate test were 100% and 25%, respectively. When the phenotypic methods for the detection of biofilm producers, namely growth on microtitre plates and Congo red agar, were compared, the sensitivity and specificity were 86% and 100%, respectively. Therefore, growth on Congo red agar and the microtitre plate test are methods that could be used to determine whether an isolate has the potential for biofilm production. PMID:24159293

  20. Plate electronics

    NASA Astrophysics Data System (ADS)

    Carlowicz, Michael

    Using a Cray T3D supercomputer and a simple assumption about the physical character of Earth's mantle, a pair of researchers from the University of California at Berkeley have built a computer model that may help explain why the planet's tectonic plates look the way they do.In creating a three-dimensional numerical simulation of convection in the Earth's interior, UC researchers Hans-Peter Bunge and Mark Richards simplified their model to account for just one major physical effect: that the viscosity of the mantle increases with depth. Reviewing some recent—but not yet widely accepted—seismic data, Bunge and Richards assumed for the sake of the model that the viscosity of the mantle increases by a factor of 30 from the lithosphere to the core-mantle boundary. Relying on that assumption, the pair ran the model for nearly three weeks on a supercomputer at Los Alamos National Laboratory and found that the simulation produced an effect similar to what we see on the surface of Earth. The model produced a surface paralleling the actual width of plates and the geometry of the plate boundaries.

  1. Formation of III–V ternary solid solutions on GaAs and GaSb plates via solid-phase substitution reactions

    SciTech Connect

    Vasil’ev, V. I.; Gagis, G. S. Kuchinskii, V. I.; Danil’chenko, V. G.

    2015-07-15

    Processes are considered in which ultrathin layers of III–V ternary solid solutions are formed via the delivery of Group-V element vapors to GaAs and GaSb semiconductor plates, with solid-phase substitution reactions occurring in the surface layers of these plates. This method can form defect-free GaAs{sup 1–x}P{sup x}, GaAs{sup x}Sb{sup 1–x}, and GaP{sup x}Sb{sup 1–x} layers with thicknesses of 10–20 nm and a content x of the embedded components of up to 0.04.

  2. The Hydrothermal System at Home Plate in Gusev Crater, Mars: Formation of High Silica Material by Acid-Sulfate Alteration of Basalt

    NASA Technical Reports Server (NTRS)

    Morris, R. V.; Ming, D. W.; Gellert, R.; Yen, A.; Clark, B. C.; Gnaff, T. G.; Arvidson, R. E.; Squyres, S. W.

    2008-01-01

    The Alpha Particle X-ray Spectrometer (APXS) instrument on the Mars Exploration Rover (MER) Spirit measured three targets on or adjacent to Home Plate in Gusev Crater that have unusually high SiO2 concentrations (68% to 91%), unusually low FeO concentrations (1% to 7%, with total Fe as FeO), and unusually high TiO2/FeO ratios (0.2 to 1.2 by weight) [1]. Two targets (Kenosha Comets and Lefty Ganote) are located on high albedo soil (Gertrude Weise) that was exposed by the rover wheels, and one target is a float rock called Fuzzy Smith. Kenosha Comets has the highest SiO2 concentration, lowest FeO concentration, and highest TiO2/FeO ratio. Mineralogical evidence from the MER Miniature Thermal Emission Spectrometer (Mini-TES) suggests that the SiO2 is present as amorphous (noncrystalline) SiO2 at Gertrude Weise and nearby targets [2,3]. Mini-TES data were not acquired for Fuzzy Smith. Home Plate is considered to have an explosive volcanic origin, resulting when basaltic magma came into contact with ground water or ice [4]. Within 50 m to 1 km of Home Plate are sulfate rich soil deposits (Paso Robles class soils with 22-35% SO3) which are considered to be probable fumarolic and/or hydrothermal deposits associated with the volcanism [5]. We develop the model here, suggested by [5], that the high-silica materials are another manifestation of acid-sulfate processes associated with fumarolic and hydrothermal activity at Home Plate. This is done by analogy with basaltic materials altered by acid sulfate processes on the Island of Hawaii.

  3. Rozhen Observatory wide-field plate archives

    NASA Astrophysics Data System (ADS)

    Tsvetkova, K.; Tsvetkov, M.

    2013-01-01

    This paper provides an overview of the wide-field (>10) plate archives at disposal in the Institute of Astronomy and National Astronomical Observatory Rozhen of the Bulgarian Academy of Sciences. The plates with total number 10093 are obtained in the period November 1978 - February 1998 with the 2 m RCC telescope and 50/70/172 cm Schmidt telescope. The available plate catalogues are upgraded with new information, as well as with link to the digitized logbooks (in JPEG file format). The digitization with high quality EPSON flatbed scanners of the available 2 m RCC telescope plates (with low resolution in JPEG, and with high resolution in standardized FITS file format) is accomplished. The digitization of the available 50/70/172 cm Schmidt telescope plates is running. The low resolution images of the scanned Rozhen plates are included in the Wide-Field Plate Database (http://wfpdb.org) and accessible online.

  4. The provenance and tectonic setting of the Lower Devonian sandstone of the Danlin Formation in southeast Yangtze Plate, with implications for the Wuyi-Yunkai orogeny in South China Block

    NASA Astrophysics Data System (ADS)

    Zhang, Jiawei; Ye, Taiping; Li, Shuangcheng; Yuan, Guohua; Dai, Chuangu; Zhang, Hui; Ma, Yibo

    2016-12-01

    The South China Block was subject to widespread tectonic and magmatic events during the middle Ordovician to earliest Devonian which are collectively called the Wuyi-Yunkai orogeny. Two different hypotheses were formulated about the origin of the orogeny: collisional orogenesis and intracontinental orogenesis. Ages of 215 detrital zircons were obtained from quartz sandstones in the Lower Devonian Danlin Formation exposed in Dushan County, south Guizhou Province. The results show that the detrital zircons came from multiple source areas but with little indication of the Wuyi-Yunkai orogeny. The detrital zircons of early Paleozoic age account only for 1.9% of all samples. These zircons range from 476 to 402 Ma and originated from the early Paleozoic granites situated east of Dushan County. The mean 206Pb/238U age of the two youngest detrital zircons was 404 Ma, which constrains the maximum depositional age of the Danlin Formation. The detrital zircons are mostly within Neoproterozoic age (59.5%) and in the range 997 to 557 Ma. The zircon age pattern, morphology, and trace element characteristics combined with paleogeographic reconstruction interpret that these detrital zircons were supplied from igneous rocks in the Neoproterozoic Sibao and Danzhou Groups within the west Jiangnan orogen to the east of the study area. Mesoproterozoic zircons made up 20.9% of the grains and range from 1569 to 1055 Ma; the ages of most of these zircons coincide with those of zircons indicating the Grenvillian orogeny (1300-1000 Ma) in the Cathaysia Plate. The detrital zircon age data reveal that the study area was located in an intracontinental tectonic setting, both before and after the Wuyi-Yunkai orogeny. Therefore, the Cathaysia Plate was not separated from the Yangtze Plate by an ocean and the study area received sediments primarily from the Cathaysia Plate located to the southeast since the middle Ordovician. By the early Devonian, the sediments were supplied by the west Jiangnan

  5. The radiolarian evidence for the accretion of the Fu-saki Formation with the inferred oceanic plate stratigraphy: A case of weakly-metamorphosed accretionary complex in Ishigaki Jima, southern Ryukyu Arc, Japan

    NASA Astrophysics Data System (ADS)

    Nakae, Satoshi

    2013-09-01

    The island of Ishigaki Jima, located in the western part of the southern Ryukyu Arc, Japan, is underlain by a basement comprising the Tumuru and Fu-saki formations. The former is a pelitic glaucophane schist with a metamorphic age of 220-190 Ma, and the latter is a weakly metamorphosed accretionary complex, composed mainly of chert, mudstone and sandstone with minor amounts of limestone and mafic rocks. The Fu-saki Formation was weakly metamorphosed at ∼140 Ma. Latest Carboniferous-Early Jurassic microfossils have been obtained from the limestones, cherts and siliceous mudstones of this formation, but no fossils have been collected from the phyllitic mudstones. The radiolarian fauna of the phyllitic mudstones described herein indicates a late Pliensbachian-early Toarcian (Early Jurassic) age. This result, when combined with existing data, enables the reconstruction of an oceanic plate stratigraphy, showing a succession of (in ascending order) Upper Carboniferous-Triassic cherts, Sinemurian-lower Pliensbachian siliceous mudstones and upper Pliensbachian-lower Toarcian phyllitic mudstones and sandstones. The radiolarians from the phyllitic mudstones are important in constraining the timing of the accretion of the Fu-saki Formation to the base of the Tumuru Formation.

  6. 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

  7. Intermittent Plate Tectonics

    NASA Astrophysics Data System (ADS)

    Silver, P. G.; Behn, M. D.

    2006-12-01

    prevent the ultimate closure of the Pacific basin and thus the cessation of subduction. More noteworthy is where subduction is not initiating. First, there is no evidence for subduction initiation anywhere within the Atlantic basin (excluding the Caribbean and Scotia), despite the mature 100-200 my age of passive-margin oceanic lithosphere. The formation of the Alpine-Himalayan chain represents the cessation of roughly 10,000 km of subduction at about 35-50 ma, Yet, no new subduction zones have initiated south of India or Africa, the two major continents that participated in the collision. These examples illustrate that subduction does not immediately initiate following a continent-continent collision, and may lag by 10s if not 100s of millions of years. The stoppage of plate tectonics, or even a dramatic reduction in subduction flux, would have significant thermal consequences for the mantle. It would effectively mark a temporary switch to "stagnant-lid" tectonics, analogous to that found on Venus, resulting in a significant increase in global mantle potential temperature (30- 100°C per 100 my) and a possibly widespread increase in magmatic activity. Such a hiatus may have occurred in the Mid-Proterozoic (1.1-1.6Ga), an era characterized by the virtual absence of orogenic activity, the longest-lived passive margin (600 My), and the production of enigmatic "anorogenic" granites found over thousands of kilometers in a belt presently stretching from southwestern to northeastern North America.

  8. MICROTUBULE-ASSOCIATED PROTEIN65 is essential for maintenance of phragmoplast bipolarity and formation of the cell plate in Physcomitrella patens.

    PubMed

    Kosetsu, Ken; de Keijzer, Jeroen; Janson, Marcel E; Goshima, Gohta

    2013-11-01

    The phragmoplast, a plant-specific apparatus that mediates cytokinesis, mainly consists of microtubules (MTs) arranged in a bipolar fashion, such that their plus ends interdigitate at the equator. Membrane vesicles are thought to move along the MTs toward the equator and fuse to form the cell plate. Although several genes required for phragmoplast MT organization have been identified, the mechanisms that maintain the bipolarity of phragmoplasts remain poorly understood. Here, we show that engaging phragmoplast MTs in a bipolar fashion in protonemal cells of the moss Physcomitrella patens requires the conserved MT cross-linking protein MICROTUBULE-ASSOCIATED PROTEIN65 (MAP65). Simultaneous knockdown of the three MAP65s expressed in those cells severely compromised MT interdigitation at the phragmoplast equator after anaphase onset, resulting in the collapse of the phragmoplast in telophase. Cytokinetic vesicles initially localized to the anaphase midzone as normal but failed to further accumulate in the next several minutes, although the bipolarity of the MT array was preserved. Our data indicate that the presence of bipolar MT arrays is insufficient for vesicle accumulation at the equator and further suggest that MAP65-mediated MT interdigitation is a prerequisite for maintenance of bipolarity of the phragmoplast and accumulation and/or fusion of cell plate-destined vesicles at the equatorial plane.

  9. 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.

  10. Multiplexed chemiluminescent assays in ArrayPlates for high-throughput measurement of gene expression

    NASA Astrophysics Data System (ADS)

    Martel, Ralph R.; Rounseville, Matthew P.; Botros, Ihab W.; Seligmann, Bruce E.

    2002-06-01

    Multiplexed Molecular Profiling (MMP) assays for drug discovery are performed in ArrayPlates. ArrayPlates are 96- well microtiter plates that contain a 16-element array at the bottom of each well. Each element within an array measures one analyte in a sample. A CCD imager records the quantitative chemiluminescent readout of all 1,536 elements in a 96-well plate simultaneously. Since array elements are reagent modifiable by the end-user, ArrayPlates can be adapted to a broad range of nucleic acid- and protein-based assays. Such multiplexed assays are rapidly established, flexible, robust, automation-friendly and cost-effective. Nucleic acid assays in ArrayPlates can detect DNA and RNA, including SNPs and ESTs. A multiplexed mRNA assay to measure the expression of 16 genes is described. The assay combines a homogeneous nuclease protection assay with subsequent probe immobilization to the array by means of a sandwich hybridization followed with chemiluminescent detection. This assay was used to examine cells grown and treated in microplates and avoided cloning, transfection, RNA insolation, reverse transcription, amplification and fluorochrome labeling. Standard deviations for the measurement of 16 genes ranged from 3 percent to 13 percent in samples of 30,000 cells. Such ArrayPlates transcription assays are useful in drug discovery and development for target validation, screening, lead optimization, metabolism and toxicity profiling. Chemiluminescent detection provides ArrayPlates assays with high signal-to-noise readout and simplifies imager requirements. Imaging a 2D surface that contains arrays simplifies lens requirements relative to imaging columns of liquid in microtiter plate wells. The Omix imager for ArrayPlates is described.

  11. Petroleum occurrences and plate tectonics

    SciTech Connect

    Olenin, V.B.; Sokolov, B.A.

    1983-01-01

    This paper analyzes the mechanisms of petroleum formation and petroleum accumulation proposed in recent years by some Russian and foreign investigators from the viewpoint of the new global or plate tectonics. On the basis of discussion and the facts, the authors conclude that the mechanisms proposed are in contradiction to reality and their use in practical application is at least premature.

  12. Corrugated cover plate for flat plate collector

    DOEpatents

    Hollands, K. G. Terry; Sibbitt, Bruce

    1978-01-01

    A flat plate radiant energy collector is providing having a transparent cover. The cover has a V-corrugated shape which reduces the amount of energy reflected by the cover away from the flat plate absorber of the collector.

  13. Plate Tectonic Cycle. K-6 Science Curriculum.

    ERIC Educational Resources Information Center

    Blueford, J. R.; And Others

    Plate Tectonics Cycle is one of the units of a K-6 unified science curriculum program. The unit consists of four organizing sub-themes: (1) volcanoes (covering formation, distribution, and major volcanic groups); (2) earthquakes (with investigations on wave movements, seismograms and sub-suface earth currents); (3) plate tectonics (providing maps…

  14. Plate Tectonic Cycle. K-6 Science Curriculum.

    ERIC Educational Resources Information Center

    Blueford, J. R.; And Others

    Plate Tectonics Cycle is one of the units of a K-6 unified science curriculum program. The unit consists of four organizing sub-themes: (1) volcanoes (covering formation, distribution, and major volcanic groups); (2) earthquakes (with investigations on wave movements, seismograms and sub-suface earth currents); (3) plate tectonics (providing maps…

  15. Cross strip anode readouts for large format, photon counting microchannel plate detectors: developing flight qualified prototypes of the detector and electronics

    NASA Astrophysics Data System (ADS)

    Vallerga, John; Raffanti, Rick; Cooney, Michael; Cumming, Harley; Varner, Gary; Seljak, Andrej

    2014-07-01

    Photon counting microchannel plate (MCP) imagers have been the detector of choice for most UV astronomical missions over the last two decades (e.g. EUVE, FUSE, COS on Hubble etc.). Over this duration, improvements in the MCP laboratory readout technology have resulted in better spatial resolution (x10), temporal resolution (x1000) and output event rate (x100), all the while operating at lower gain (x 10) resulting in lower high voltage requirements and longer MCP lifetimes. One such technology is the parallel cross strip (PXS) readout. Laboratory versions of PXS electronics have demonstrated < 20 μm FWHM spatial resolution, count rates on the order of 2 MHz, and temporal resolution of ~ 1ns. In 2012 our group at U.C. Berkeley, along with our partners at the U. Hawaii, received a Strategic Astrophysics Technology grant to raise the TRL of the PXS detector and electronics from 4 to 6 by replacing most of the high powered electronics with application specific integrated circuits (ASICs) which will lower the power, mass and volume requirements of the PXS detector. We were also tasked to design and fabricate a "standard" 50mm square active area MCP detector incorporating these electronics that can be environmentally qualified for flight (temperature, vacuum, vibration). The first ASICs designed for this program have been fabricated and are undergoing testing. We present the latest progress on these ASIC designs and performance and show imaging results from the new 50 x 50 mm XS detector.

  16. Optimization of tetrazolium salt assay for Pseudomonas aeruginosa biofilm using microtiter plate method.

    PubMed

    Sabaeifard, Parastoo; Abdi-Ali, Ahya; Soudi, Mohammad Reza; Dinarvand, Rasoul

    2014-10-01

    Pseudomonas aeruginosa is one of the most important pathogenic bacteria related to biofilm infections. Due to the biofilm multi-drug resistance, methods of biofilm formation enumeration are of interest for assessment of efficient drug regimen development for biofilm inhibition or eradication. There are many different assay methods to determine the biofilm formation, using vital or non-vital dyes. The primary aim of the current study was to develop an assay using a member of tetrazolium salts family, 2,3,5-triphenyl-tetrazolium chloride (TTC), for detection of P. aeruginosa biofilm formation in 96-well microtiter plates and also a method of Minimum Biofilm Inhibitory Concentration (MBIC) determination of antibiotics against P. aeruginosa PAO1. Furthermore, the assay was optimized for TTC concentration, wavelength and period of incubation for 4 different antibiotics. The optimized condition was then compared with two other prevalent methods: the crystal violet (CV) assay and the 2,3-bis (2-methoxy-4-nitro-5-sulfophenly)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) assay. In general, the optimized TTC assay (0.5% TTC, 6h of incubation and absorbance measurement at 405nm for biofilm assay and 1% TTC, 5h of incubation and absorbance measurement at 490nm for MBIC determination) distinguished between biofilms formed by different concentrations of bacteria and also was able to detect lower amounts of biofilm formed in contrast to the other two assay methods suggesting that TTC assay is more sensitive and also less expensive than other vital staining methods. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Formation and evolution of a metasomatized lithospheric root at the motionless Antarctic plate: the case of East Island, Crozet Archipelago (Indian Ocean)

    NASA Astrophysics Data System (ADS)

    Meyzen, Christine; Marzoli, Andrea; Bellieni, Giuliano; Levresse, Gilles

    2016-04-01

    Sitting atop the nearly stagnant Antarctic plate (ca. 6.46 mm/yr), the Crozet archipelago midway between Madagascar and Antarctica constitutes a region of unusually shallow (1543-1756 m below sea level) and thickened oceanic crust (10-16.5 km), high geoid height, and deep low-velocity zone, which may reflect the surface expression of a mantle plume. Here, we present new major and trace element data for Quaternary sub-aerial alkali basalts from East Island, the easterly and oldest island (ca. 9 Ma) of the Crozet archipelago. Crystallization at uppermost mantle depth and phenocryst accumulation have strongly affected their parental magma compositions. Their trace element patterns show a large negative K anomaly relative to Ta-La, moderate depletions in Rb and Ba with respect to Th-U, and heavy rare earth element (HREE) depletions relative to light REE. These characteristics allow limits to be placed upon the composition and mineralogy of their mantle source. The average trace element spectrum of East Island basalts can be matched by melting of about 2 % of a garnet-phlogopite-bearing peridotite source. The stability field of phlogopite restricts melting depth to lithospheric levels. The modelled source composition requires a multistage evolution, where the mantle has been depleted by melt extraction before having been metasomatized by alkali-rich plume melts. The depleted mantle component may be sourced by residual mantle plume remnants stagnated at the melting locus due to a weak lateral flow velocity inside the melting regime, whose accumulation progressively edifies a depleted lithospheric root above the plume core. Low-degree alkali-rich melts are likely derived from the plume source. Such a mantle source evolution may be general to both terrestrial and extraterrestrial environments where the lateral component velocity of the mantle flow field is extremely slow.

  18. Nitric oxide regulates neutrophil migration through microparticle formation.

    PubMed

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

    2008-01-01

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

  19. Delamination, upper plate extension, and plate margin complexity

    NASA Astrophysics Data System (ADS)

    Ueda, Kosuke; Gerya, Taras; Willett, Sean

    2017-04-01

    We investigate the syn- and post-subduction margin evolution with respect to extension, lithospheric removal, and magmatic and topographic consequences by employing 3D geodynamic models. In all experiments, regions of extended partial melting are overlain by up to 3 km high plateaus. There is complex geometric entanglement between upper mantle, partially molten rocks, and lithosphere, which is thermally eroded, over hundreds of kilometers across the plate contact. A complex lithosphere-asthenosphere-boundary features elongated anomalies at scales of few tens to hundred kilometers. First-order, synthetic seismic anomaly patterns, based on thermodynamic velocities which are tabulated for model p,T conditions, are accordingly complex. Passive margin geometry variations in the lower plate effect consistent and inherited differences in dynamic evolution. Promontories along the margin tend to trigger three stages of evolution: 1) a magmatic arc; 2) a lower plate, eduction-like exhumation of buried continental crust in domal patterns of few tens of km wavelength; and subsequently, 3) the formation of extended zones on the upper plate which lack a lithospheric mantle, undergo partial extension, and feature lower crustal melting. Slab break-off is consistently favoured in locations where the lower plate margin is relatively recessed. Concerning the classical removal mechanisms, transitions and co-evolution between delamination, convective thinning, and upper-plate extension are gradual and these modes are not mutually exclusive. Almost complete mixed-mode removal and extension can be compared to the Aegean. Slab window formation by margin geometry variation produces characteristic uplift patterns that are comparable to the Apennines, where higher uplift rates could be a consequence of incipient necking of the slab below central Calabria.

  20. Formation of randomly distributed nano-tubes, -rods and -plates of n-type and p-type bismuth telluride via molecular legation

    NASA Astrophysics Data System (ADS)

    Ram, Jasa; Ghosal, Partha

    2015-08-01

    Randomly distributed nanotubes, nanorods and nanoplates of Bi0.5Sb1.5Te3 and Bi2Te2.7Se0.3 ternary compounds have been synthesized via a high yield solvo-thermal process. Prior to solvo-thermal heating at 230 °C for crystallization, we ensured molecular legation in room temperature reaction by complete reduction of precursor materials, dissolved in ethylene glycol and confirmed it by replicating Raman spectra of amorphous and crystalline materials. These nanomaterials have also been characterized using XRD, FE-SEM, EDS and TEM. Possible formation mechanism is also discussed. This single process will enable development of thermoelectric modules and random distribution of diverse morphology will be beneficial in retaining nano-crystallite sizes.

  1. Sputtering and ion plating

    NASA Technical Reports Server (NTRS)

    1972-01-01

    The proceedings of a conference on sputtering and ion plating are presented. Subjects discussed are: (1) concepts and applications of ion plating, (2) sputtering for deposition of solid film lubricants, (3) commercial ion plating equipment, (4) industrial potential for ion plating and sputtering, and (5) fundamentals of RF and DC sputtering.

  2. Methods for inducing embryoid body formation: in vitro differentiation system of embryonic stem cells.

    PubMed

    Kurosawa, Hiroshi

    2007-05-01

    When cultured in suspension without antidifferentiation factors, embryonic stem (ES) cells spontaneously differentiate and form three-dimensional multicellular aggregates called embryoid bodies (EBs). EBs recapitulate many aspects of cell differentiation during early embryogenesis, and play an important role in the differentiation of ES cells into a variety of cell types in vitro. There are several methods for inducing the formation of EBs from ES cells. The three basic methods are liquid suspension culture in bacterial-grade dishes, culture in methylcellulose semisolid media, and culture in hanging drops. Recently, the methods using a round-bottomed 96-well plate and a conical tube are adopted for forming EBs from predetermined numbers of ES cells. For the production of large numbers of EBs, stirred-suspension culture using spinner flasks and bioreactors is performed. Each of these methods has its own peculiarity; thus, the features of formed EBs depending on the method used. Therefore, we should choose an appropriate method for EB formation according to the objective to be attained. In this review, we summarize the studies on in vitro differentiation of ES cells via EB formation and highlight the EB formation methods recently developed including the techniques, devices, and procedures involved.

  3. The cytological changes of tobacco zygote and proembryo cells induced by beta-glucosyl Yariv reagent suggest the involvement of arabinogalactan proteins in cell division and cell plate formation

    PubMed Central

    2012-01-01

    Background In dicotyledonous plant, the first asymmetric zygotic division and subsequent several cell divisions are crucial for proembryo pattern formation and later embryo development. Arabinogalactan proteins (AGPs) are a family of extensively glycosylated cell surface proteins that are thought to have important roles in various aspects of plant growth and development, including embryogenesis. Previous results from our laboratory show that AGPs are concerned with tobacco egg cell fertilization and zygotic division. However, how AGPs interact with other factors involved in zygotic division and proembryo development remains unknown. Results In this study, we used the tobacco in vitro zygote culture system and series of meticulous cell biology techniques to investigate the roles of AGPs in zygote and proembryo cell division. For the first time, we examined tobacco proembryo division patterns detailed to every cell division. The bright-field images and statistical results both revealed that with the addition of an exogenous AGPs inhibitor, beta-glucosyl Yariv (beta-GlcY) reagent, the frequency of aberrant division increased remarkably in cultured tobacco zygotes and proembryos, and the cell plate specific locations of AGPs were greatly reduced after beta-GlcY treatment. In addition, the accumulations of new cell wall materials were also significantly affected by treating with beta-GlcY. Detection of cellulose components by Calcofluor white stain showed that strong fluorescence was located in the newly formed wall of daughter cells after the zygotic division of in vivo samples and the control samples from in vitro culture without beta-GlcY treatment; while there was only weak fluorescence in the newly formed cell walls with beta-GlcY treatment. Immunocytochemistry examination with JIM5 and JIM7 respectively against the low- and high-esterified pectins displayed that these two pectins located in opposite positions of zygotes and proembryos in vivo and the polarity was

  4. The cytological changes of tobacco zygote and proembryo cells induced by beta-glucosyl Yariv reagent suggest the involvement of arabinogalactan proteins in cell division and cell plate formation.

    PubMed

    Yu, Miao; Zhao, Jie

    2012-08-01

    In dicotyledonous plant, the first asymmetric zygotic division and subsequent several cell divisions are crucial for proembryo pattern formation and later embryo development. Arabinogalactan proteins (AGPs) are a family of extensively glycosylated cell surface proteins that are thought to have important roles in various aspects of plant growth and development, including embryogenesis. Previous results from our laboratory show that AGPs are concerned with tobacco egg cell fertilization and zygotic division. However, how AGPs interact with other factors involved in zygotic division and proembryo development remains unknown. In this study, we used the tobacco in vitro zygote culture system and series of meticulous cell biology techniques to investigate the roles of AGPs in zygote and proembryo cell division. For the first time, we examined tobacco proembryo division patterns detailed to every cell division. The bright-field images and statistical results both revealed that with the addition of an exogenous AGPs inhibitor, beta-glucosyl Yariv (beta-GlcY) reagent, the frequency of aberrant division increased remarkably in cultured tobacco zygotes and proembryos, and the cell plate specific locations of AGPs were greatly reduced after beta-GlcY treatment. In addition, the accumulations of new cell wall materials were also significantly affected by treating with beta-GlcY. Detection of cellulose components by Calcofluor white stain showed that strong fluorescence was located in the newly formed wall of daughter cells after the zygotic division of in vivo samples and the control samples from in vitro culture without beta-GlcY treatment; while there was only weak fluorescence in the newly formed cell walls with beta-GlcY treatment. Immunocytochemistry examination with JIM5 and JIM7 respectively against the low- and high-esterified pectins displayed that these two pectins located in opposite positions of zygotes and proembryos in vivo and the polarity was not affected by

  5. The Postcranial Skeleton of an Exceptionally Complete Individual of the Plated Dinosaur Stegosaurus stenops (Dinosauria: Thyreophora) from the Upper Jurassic Morrison Formation of Wyoming, U.S.A.

    PubMed

    Maidment, Susannah Catherine Rose; Brassey, Charlotte; Barrett, Paul Michael

    2015-01-01

    Although Stegosaurus is one of the most iconic dinosaurs, well-preserved fossils are rare and as a consequence there is still much that remains unknown about the taxon. A new, exceptionally complete individual affords the opportunity to describe the anatomy of Stegosaurus in detail for the first time in over a century, and enables additional comparisons with other stegosaurian dinosaurs. The new specimen is from the Red Canyon Ranch Quarry, near Shell Wyoming, and appears to have been so well preserved because it was buried rapidly in a pond or body of standing water immediately after death. The quarry is probably located in the middle part of the Morrison Formation, which is believed to be Tithonian in age in this area. The specimen is referable to Stegosaurus stenops based on the possession of an edentulous anterior portion of the dentary and elevated postzygapophyses on the cervical vertebrae. New information provided by the specimen concerns the morphology of the vertebrae, the iliosacral block and dermal armor. Several aspects of its morphology indicate the individual was not fully skeletally mature at the time of death, corroborating a previous histological study.

  6. The Postcranial Skeleton of an Exceptionally Complete Individual of the Plated Dinosaur Stegosaurus stenops (Dinosauria: Thyreophora) from the Upper Jurassic Morrison Formation of Wyoming, U.S.A.

    PubMed Central

    Maidment, Susannah Catherine Rose; Brassey, Charlotte; Barrett, Paul Michael

    2015-01-01

    Although Stegosaurus is one of the most iconic dinosaurs, well-preserved fossils are rare and as a consequence there is still much that remains unknown about the taxon. A new, exceptionally complete individual affords the opportunity to describe the anatomy of Stegosaurus in detail for the first time in over a century, and enables additional comparisons with other stegosaurian dinosaurs. The new specimen is from the Red Canyon Ranch Quarry, near Shell Wyoming, and appears to have been so well preserved because it was buried rapidly in a pond or body of standing water immediately after death. The quarry is probably located in the middle part of the Morrison Formation, which is believed to be Tithonian in age in this area. The specimen is referable to Stegosaurus stenops based on the possession of an edentulous anterior portion of the dentary and elevated postzygapophyses on the cervical vertebrae. New information provided by the specimen concerns the morphology of the vertebrae, the iliosacral block and dermal armor. Several aspects of its morphology indicate the individual was not fully skeletally mature at the time of death, corroborating a previous histological study. PMID:26466098

  7. Scale-up from microtiter plate to laboratory fermenter: evaluation by online monitoring techniques of growth and protein expression in Escherichia coli and Hansenula polymorpha fermentations

    PubMed Central

    2009-01-01

    Background In the past decade, an enormous number of new bioprocesses have evolved in the biotechnology industry. These bioprocesses have to be developed fast and at a maximum productivity. Up to now, only few microbioreactors were developed to fulfill these demands and to facilitate sample processing. One predominant reaction platform is the shaken microtiter plate (MTP), which provides high-throughput at minimal expenses in time, money and work effort. By taking advantage of this simple and efficient microbioreactor array, a new online monitoring technique for biomass and fluorescence, called BioLector, has been recently developed. The combination of high-throughput and high information content makes the BioLector a very powerful tool in bioprocess development. Nevertheless, the scalabilty of results from the micro-scale to laboratory or even larger scales is very important for short development times. Therefore, engineering parameters regarding the reactor design and its operation conditions play an important role even on a micro-scale. In order to evaluate the scale-up from a microtiter plate scale (200 μL) to a stirred tank fermenter scale (1.4 L), two standard microbial expression systems, Escherichia coli and Hansenula polymorpha, were fermented in parallel at both scales and compared with regard to the biomass and protein formation. Results Volumetric mass transfer coefficients (kLa) ranging from 100 to 350 1/h were obtained in 96-well microtiter plates. Even with a suboptimal mass transfer condition in the microtiter plate compared to the stirred tank fermenter (kLa = 370-600 1/h), identical growth and protein expression kinetics were attained in bacteria and yeast fermentations. The bioprocess kinetics were evaluated by optical online measurements of biomass and protein concentrations exhibiting the same fermentation times and maximum signal deviations below 10% between the scales. In the experiments, the widely applied green fluorescent protein (GFP

  8. Dynamic Stabilization of Simple Fractures With Active Plates Delivers Stronger Healing Than Conventional Compression Plating

    PubMed Central

    Tsai, Stanley; Bliven, Emily K.; von Rechenberg, Brigitte; Kindt, Philipp; Augat, Peter; Henschel, Julia; Fitzpatrick, Daniel C.; Madey, Steven M.

    2017-01-01

    Objectives: Active plates dynamize a fracture by elastic suspension of screw holes within the plate. We hypothesized that dynamic stabilization with active plates delivers stronger healing relative to standard compression plating. Methods: Twelve sheep were randomized to receive either a standard compression plate (CP) or an active plate (ACTIVE) for stabilization of an anatomically reduced tibial osteotomy. In the CP group, absolute stabilization was pursued by interfragmentary compression with 6 cortical screws. In the ACTIVE group, dynamic stabilization after bony apposition was achieved with 6 elastically suspended locking screws. Fracture healing was analyzed weekly on radiographs. After sacrifice 9 weeks postsurgery, the torsional strength of healed tibiae and contralateral tibiae was measured. Finally, computed tomography was used to assess fracture patterns and healing modes. Results: Healing in both groups included periosteal callus formation. ACTIVE specimens had almost 6 times more callus area by week 9 (P < 0.001) than CP specimens. ACTIVE specimens recovered on average 64% of their native strength by week 9, and were over twice as strong as CP specimens, which recovered 24% of their native strength (P = 0.008). Microcomputed tomography demonstrated that compression plating induced a combination of primary bone healing and gap healing. Active plating consistently stimulated biological bone healing by periosteal callus formation. Conclusions: Compared with compression plating, dynamic stabilization of simple fractures with active plates delivers significantly stronger healing. PMID:27861456

  9. Survey of ion plating sources

    NASA Technical Reports Server (NTRS)

    Spalvins, T.

    1979-01-01

    Ion plating is a plasma deposition technique where ions of the gas and the evaporant have a decisive role in the formation of a coating in terms of adherence, coherence, and morphological growth. The range of materials that can be ion plated is predominantly determined by the selection of the evaporation source. Based on the type of evaporation source, gaseous media and mode of transport, the following will be discussed: resistance, electron beam sputtering, reactive and ion beam evaporation. Ionization efficiencies and ion energies in the glow discharge determine the percentage of atoms which are ionized under typical ion plating conditions. The plating flux consists of a small number of energetic ions and a large number of energetic neutrals. The energy distribution ranges from thermal energies up to a maximum energy of the discharge. The various reaction mechanisms which contribute to the exceptionally strong adherence - formation of a graded substrate/coating interface are not fully understood, however the controlling factors are evaluated. The influence of process variables on the nucleation and growth characteristics are illustrated in terms of morphological changes which affect the mechanical and tribological properties of the coating.

  10. Failure analysis of stainless steel femur fixation plate.

    PubMed

    Hussain, P B; Mohammad, M

    2004-05-01

    Failure analysis was performed to investigate the failure of the femur fixation plate which was previously fixed on the femur of a girl. Radiography, metallography, fractography and mechanical testing were conducted in this study. The results show that the failure was due to the formation of notches on the femur plate. These notches act as stress raisers from where the cracks start to propagate. Finally fracture occurred on the femur plate and subsequently, the plate failed.

  11. Williamson Polishing & Plating Site

    EPA Pesticide Factsheets

    Williamson Polishing & Plating Co. Inc. was a plating shop located in the Martindale-Brightwood neighborhood of Indianapolis. The facility conducted job shop polishing and electroplating services. The vacant site contains a 14,651-square-foot building.

  12. Growth Plate Fractures

    MedlinePlus

    ... the most widely used by doctors is the Salter-Harris system, described below. Type I Fractures These ... incidence of growth plate fractures peaks in adolescence. Salter-Harris classification of growth plate fractures. AAOS does ...

  13. Earth's Decelerating Tectonic Plates

    SciTech Connect

    Forte, A M; Moucha, R; Rowley, D B; Quere, S; Mitrovica, J X; Simmons, N A; Grand, S P

    2008-08-22

    Space geodetic and oceanic magnetic anomaly constraints on tectonic plate motions are employed to determine a new global map of present-day rates of change of plate velocities. This map shows that Earth's largest plate, the Pacific, is presently decelerating along with several other plates in the Pacific and Indo-Atlantic hemispheres. These plate decelerations contribute to an overall, globally averaged slowdown in tectonic plate speeds. The map of plate decelerations provides new and unique constraints on the dynamics of time-dependent convection in Earth's mantle. We employ a recently developed convection model constrained by seismic, geodynamic and mineral physics data to show that time-dependent changes in mantle buoyancy forces can explain the deceleration of the major plates in the Pacific and Indo-Atlantic hemispheres.

  14. Accelerated plate tectonics.

    PubMed

    Anderson, D L

    1975-03-21

    The concept of a stressed elastic lithospheric plate riding on a viscous asthenosphere is used to calculate the recurrence interval of great earthquakes at convergent plate boundaries, the separation of decoupling and lithospheric earthquakes, and the migration pattern of large earthquakes along an arc. It is proposed that plate motions accelerate after great decoupling earthquakes and that most of the observed plate motions occur during short periods of time, separated by periods of relative quiescence.

  15. Plating Tank Control Software

    SciTech Connect

    Krafcik, John

    1998-03-01

    The Plating Tank Control Software is a graphical user interface that controls and records plating process conditions for plating in high aspect ratio channels that require use of low current and long times. The software is written for a Pentium II PC with an 8 channel data acquisition card, and the necessary shunt resistors for measuring currents in the millampere range.

  16. Rotatable shear plate interferometer

    DOEpatents

    Duffus, Richard C.

    1988-01-01

    A rotatable shear plate interferometer comprises a transparent shear plate mounted obliquely in a tubular supporting member at 45.degree. with respect to its horizontal center axis. This tubular supporting member is supported rotatably around its center axis and a collimated laser beam is made incident on the shear plate along this center axis such that defocus in different directions can be easily measured.

  17. Plate-induced Miocene extension in southern California

    SciTech Connect

    Stuart, W.D. Univ. of California, Santa Barbara, CA )

    1992-01-01

    Miocene crustal extension in southern California can be explained by the interaction of tectonic plates in relative motion. The Pacific, Juan de Fuca, and Farallon (Guadalupe) plates are represented by flat elastic plates surrounded by an infinite elastic plate, the eastern part of which represents the North America plate. Forcing is by assigned subduction pull, and tractions at all plate boundaries satisfy a viscous constitutive law. Plate bottoms are stress-free. In the first part of the solution plate velocities and boundary tractions are found from static equilibrium. Then principal horizontal stresses and strains in plate interiors caused by tractions and subduction pull are found by a boundary element procedure. Using plate boundary geometry from Stock and Hodges for early- and mid-Miocene times, it is found that the portion of the North America plate margin between the Mendocino and Rivera triple junctions has maximum extensional strain directed westward. This result is generally consistent with directions associated with metamorphic core complex formation in southern California. The model is also consistent with extensional strain and rotation sense of crustal blocks in the vicinity of Los Angeles, as inferred by Luyendyk and others from paleomagnetic data. In the model the greatest extensional strain of the North America plate occurs near the Pacific-North America transform, in the area above the absent Farallon slab. Extension direction varies from northwest to southwest according to plate geometry, subduction pull (Juan de Fuca and Guadalupe), and plate boundary tractions.

  18. Plate Tectonics, Geographical Information System, paleogeography

    SciTech Connect

    Moore, Thomas L.; Scotese, Christopher

    2002-05-24

    The PaleoX.framwork is a dynamically linked/loaded framework for Cocoa applications. The primary goal of this library is to standardize several elements used for working with paleogeographic data. This includes objects designed to organize information for tectonic plates, including maps, rotation objects, plate names, and designations. In addition, PaleoX provides object-oriented solutions for handling standard paleogeographic file formats from the PALEOMAP Project.

  19. Malachite green photosensitive plates.

    PubMed

    Solano, C

    1989-08-15

    An experimental study of the behavior of malachite green sensitized plates was carried out. The transmittance variation of the irradiated plates was taken as a parameter. It has been observed that photoreduction in the malachite green plates is present only when ammonium dichromate is added to the plates. The introduction of external electron donors does not improve the photochemical reaction. It has been determined that malachite green molecules form a weak complex with the dichromate molecules and this complex can only be destroyed photochemically. This effect can explain the limited response of the malachite green dichromated plates.

  20. An improved plating process

    NASA Technical Reports Server (NTRS)

    Askew, John C.

    1994-01-01

    An alternative to the immersion process for the electrodeposition of chromium from aqueous solutions on the inside diameter (ID) of long tubes is described. The Vessel Plating Process eliminates the need for deep processing tanks, large volumes of solutions, and associated safety and environmental concerns. Vessel Plating allows the process to be monitored and controlled by computer thus increasing reliability, flexibility and quality. Elimination of the trivalent chromium accumulation normally associated with ID plating is intrinsic to the Vessel Plating Process. The construction and operation of a prototype Vessel Plating Facility with emphasis on materials of construction, engineered and operational safety and a unique system for rinse water recovery are described.

  1. Angular shear plate

    DOEpatents

    Ruda, Mitchell C [Tucson, AZ; Greynolds, Alan W [Tucson, AZ; Stuhlinger, Tilman W [Tucson, AZ

    2009-07-14

    One or more disc-shaped angular shear plates each include a region thereon having a thickness that varies with a nonlinear function. For the case of two such shear plates, they are positioned in a facing relationship and rotated relative to each other. Light passing through the variable thickness regions in the angular plates is refracted. By properly timing the relative rotation of the plates and by the use of an appropriate polynomial function for the thickness of the shear plate, light passing therethrough can be focused at variable positions.

  2. MerMade: An Oligodeoxyribonucleotide Synthesizer for High Throughput Oligonucleotide Production in Dual 96-Well Plates

    PubMed Central

    Rayner, Simon; Brignac, Stafford; Bumeister, Ron; Belosludtsev, Yuri; Ward, Travis; Grant, O’dell; O’Brien, Kevin; Evans, Glen A.; Garner, Harold R.

    1998-01-01

    We have designed and constructed a machine that synthesizes two standard 96-well plates of oligonucleotides in a single run using standard phosphoramidite chemistry. The machine is capable of making a combination of standard, degenerate, or modified oligos in a single plate. The run time is typically 17 hr for two plates of 20-mers and a reaction scale of 40 nm. The reaction vessel is a standard polypropylene 96-well plate with a hole drilled in the bottom of each well. The two plates are placed in separate vacuum chucks and mounted on an xy table. Each well in turn is positioned under the appropriate reagent injection line and the reagent is injected by switching a dedicated valve. All aspects of machine operation are controlled by a Macintosh computer, which also guides the user through the startup and shutdown procedures, provides a continuous update on the status of the run, and facilitates a number of service procedures that need to be carried out periodically. Over 25,000 oligos have been synthesized for use in dye terminator sequencing reactions, polymerase chain reactions (PCRs), hybridization, and RT–PCR. Oligos up to 100 bases in length have been made with a coupling efficiency in excess of 99%. These machines, working in conjunction with our oligo prediction code are particularly well suited to application in automated high throughput genomic sequencing. PMID:9685322

  3. Multicolor printing plate joining

    NASA Technical Reports Server (NTRS)

    Waters, W. J. (Inventor)

    1984-01-01

    An upper plate having ink flow channels and a lower plate having a multicolored pattern are joined. The joining is accomplished without clogging any ink flow paths. A pattern having different colored parts and apertures is formed in a lower plate. Ink flow channels each having respective ink input ports are formed in an upper plate. The ink flow channels are coated with solder mask and the bottom of the upper plate is then coated with solder. The upper and lower plates are pressed together at from 2 to 5 psi and heated to a temperature of from 295 F to 750 F or enough to melt the solder. After the plates have cooled and the pressure is released, the solder mask is removed from the interior passageways by means of a liquid solvent.

  4. Fluorescence assay for mitochondrial permeability transition in cardiomyocytes cultured in a microtiter plate.

    PubMed

    Christensen, Marie Louise Muff; Braunstein, Thomas Hartig; Treiman, Marek

    2008-07-01

    Mitochondrial permeability transition pore (MPTP) is a voltage-dependent, large-conductance channel of the inner mitochondrial membrane with an important role in a range of pathophysiological conditions. To facilitate studies of pharmacological pore modulation, we describe an assay in a model using neonatal cardiomyocytes in a 96-well microtiter plate format. In the presence of mitochondrial membrane potential Delta Psi m, accumulation of rhodamine-123 in mitochondria (40,000 cells/well, 2.6 microM rhodamine-123) caused fluorescence signal quenching. Following substitution of dye-free buffer, dequenching occurred on the distribution of rhodamine-123 into the extracellular volume. The addition of a small buffer volume containing digitonin (final concentration 10 microg/ml) and Ca(2+) (final concentrations up to 100 microM free Ca(2+)) caused dequenching (Delta F) due to Delta Psi m dissipation by MPTP, as evidenced by inhibition in the presence of cyclosporin A (0.2-2 microM) and facilitation by pH 6.2. Delta F due to Delta Psi m-dissipating agent carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) or alamethicin (10 microM) was insensitive to either pH or cyclosporin A. Inhibition of Ca(2+)-induced (but not of FCCP- or alamethicin-induced) Delta F by glycogen synthase kinase 3beta (GSK3 beta) antagonist SB216763 and adenosine, acting at the level of intracellular signaling and plasma membrane receptors, respectively, is shown to illustrate potential applications of this assay. Limitation of the assay to cells with energized mitochondria is stressed.

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

    PubMed Central

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

    2015-01-01

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

  6. Characterization of antibody binding to cell surface antigens using a plasma membrane-bound plate assay.

    PubMed

    Vater, C A; Reid, K; Bartle, L M; Goldmacher, V S

    1995-01-01

    A procedure has been developed for measuring antibody binding to cell surface antigens using an immobilized plasma membrane fraction. In this method, isolated plasma membranes are dried onto wells of a 96-well microtiter plate and incubated with antibodies that recognize a cell surface protein. Bound antibody is detected indirectly using an enzyme-linked or fluorescently tagged second antibody. Alternatively, the primary antibody itself can be labeled and its binding can be detected directly. The assay is simple and fast and provides several advantages over whole cell binding assays currently in widespread use.

  7. GOLD PLATING PROCESS

    DOEpatents

    Seegmiller, R.

    1957-08-01

    An improved bath is reported for plating gold on other metals. The composition of the plating bath is as follows: Gold cyanide from about 15 to about 50 grams, potassium cyanide from about 70 to about 125 grams, and sulfonated castor oil from about 0.1 to about 10 cc. The gold plate produced from this bath is smooth, semi-hard, and nonporous.

  8. Plating methods, a survey

    NASA Technical Reports Server (NTRS)

    Berkowitz, J. B.; Emerson, N. H.

    1972-01-01

    Results are presented of a comprehensive search of the literature available, much of which has been generated by the research centers of NASA and its contractors, on plating and coating methods and techniques. Methods covered included: (1) electroplating from aqueous solutions; (2) electroplating from nonaqueous solutions; (3) electroplating from fused-salt baths; (4) electroforming; (5) electroless plating, immersion plating, and mirroring; (6) electroplating from gaseous plasmas; and (7) anodized films and conversion coatings.

  9. Tectonic Evolution of the Jurassic Pacific Plate

    NASA Astrophysics Data System (ADS)

    Nakanishi, M.; Ishihara, T.

    2015-12-01

    We present the tectonic evolution of the Jurassic Pacific plate based on magnetic anomly lineations and abyssal hills. The Pacific plate is the largest oceanic plate on Earth. It was born as a microplate aroud the Izanagi-Farallon-Phoenix triple junction about 192 Ma, Early Jurassic [Nakanishi et al., 1992]. The size of the Pacific plate at 190 Ma was nearly half that of the present Easter or Juan Fernandez microplates in the East Pacific Rise [Martinez et at, 1991; Larson et al., 1992]. The plate boundary surrounding the Pacific plate from Early Jurassic to Early Cretaceous involved the four triple junctions among Pacific, Izanagi, Farallon, and Phoenix plates. The major tectonic events as the formation of oceanic plateaus and microplates during the period occurred in the vicinity of the triple junctions [e.g., Nakanishi and Winterer, 1998; Nakanishi et al., 1999], implying that the study of the triple junctions is indispensable for understanding the tectonic evolution of the Pacific plate. Previous studies indicate instability of the configuration of the triple junctions from Late Jurassic to Early Cretaceous (155-125 Ma). On the other hand, the age of the birth of the Pacific plate was determined assuming that all triple junctions had kept their configurations for about 30 m.y. [Nakanishi et al., 1992] because of insufficient information of the tectonic history of the Pacific plate before Late Jurassic.Increase in the bathymetric and geomagnetic data over the past two decades enables us to reveal the tectonic evolution of the Pacific-Izanagi-Farallon triple junction before Late Jurassic. Our detailed identication of magnetic anomaly lineations exposes magnetic bights before anomaly M25. We found the curved abyssal hills originated near the triple junction, which trend is parallel to magnetic anomaly lineations. These results imply that the configuration of the Pacific-Izanagi-Farallon triple junction had been RRR before Late Jurassic.

  10. PLATES WITH OXIDE INSERTS

    DOEpatents

    West, J.M.; Schumar, J.F.

    1958-06-10

    Planar-type fuel assemblies for nuclear reactors are described, particularly those comprising fuel in the oxide form such as thoria and urania. The fuel assembly consists of a plurality of parallel spaced fuel plate mennbers having their longitudinal side edges attached to two parallel supporting side plates, thereby providing coolant flow channels between the opposite faces of adjacent fuel plates. The fuel plates are comprised of a plurality of longitudinally extending tubular sections connected by web portions, the tubular sections being filled with a plurality of pellets of the fuel material and the pellets being thermally bonded to the inside of the tubular section by lead.

  11. CALUTRON FACE PLATE

    DOEpatents

    Brobeck, W.M.

    1959-08-25

    The construction of a removable cover plate for a calutron tank is described. The plate is fabricated of a rectangular frame member to which is welded a bowed or dished plate of thin steel, reinforced with transverse stiffening ribs. When the tank is placed between the poles of a magnet, the plate may be pivoted away from the tank and magnet and is adapted to support the ion separation mechanism secured to its inner side as well as the vacuum load within the tank.

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

    PubMed

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

    2014-11-28

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

  13. Performance evaluation of the R6R018 fuel plate using PLATE code

    SciTech Connect

    Pavel G. Medvedev; Hakan Ozaltun

    2010-03-01

    The paper presents results of performance evaluation of the R6R018 fuel plate using PLATE code. R6R018 is a U-7Mo dispersion type mini-plate with Al-3.5Si matrix irradiated in the RERTR-9B experiment. The design of this plate is prototypical of the planned LEONIDAS irradiation test. Therefore, a detailed performance analysis of this plate is important to confirm acceptable behavior in pile, and to provide baseline and justification for further analysis and testing. Specific results presented in the paper include fuel temperature history, growth of the interaction layer between the U-Mo and the matrix, swelling, growth of the corrosion layer, and degradation of thermal conductivity. The methodology of the analysis will be discussed including the newly developed capability to account for the formation of the interaction layer during fuel fabrication.

  14. The INASAN Zvenigorod Observatory Plate Collection

    NASA Astrophysics Data System (ADS)

    Dluzhnevskaya, O. B.; Vereshchagin, S. V.; Chupina, N. V.; Osipenko, V. P.

    2012-01-01

    Description of the astronomical wide-field plate- and film-collection of the Zvenigorod Observatory of the Institute of Astronomy of the Russian Academy of Sciences is presented. The archives of the collection are the result of photographic observations conducted for more than thirty years from 1972 to 2005. They included: 1) photographic plates obtained with the 40 cm Carl Zeiss astrograph (the half of the plates are included in the Sofia WFPDB). The plates are now in process of digitization in FITS-formats, 700 MB each scan. Two EPSON 1640XL scanners are used. The observational programs include the FON photographic north sky survey, small bodies in the Solar system - asteroids and comets; 2) film archive from the AFU-75 camera; 3) archive of artificial satellites monitoring observations with wide-field camera VAU.

  15. Plating Inc. Site

    EPA Pesticide Factsheets

    EPA will begin a hazardous waste cleanup at Plating Inc., an abandoned zinc and chromate plating facility at 888 N. Prior Ave. in St. Paul. The abandoned facility houses approximately 82 open chemical vats containing hazardous wastes such as cyanide, caust

  16. Earthquakes and plate tectonics

    USGS Publications Warehouse

    Spall, H.

    1977-01-01

    An explanation is to be found in plate tectonics, a concept which has revolutionized thinking in the Earth sciences in the last 10 years. The theory of plate tectonics combines many of the ideas about continental drift (originally proposed in 1912 by Alfred Wegener in Germany) and sea-floor spreading (suggested originally by Harry Hess of Princeton University). 

  17. Blue Willow Story Plates

    ERIC Educational Resources Information Center

    Fontes, Kris

    2009-01-01

    In the December 1997 issue of "SchoolArts" is a lesson titled "Blue Willow Story Plates" by Susan Striker. In this article, the author shares how she used this lesson with her middle-school students many times over the years. Here, she describes a Blue Willow plate painting project that her students made.

  18. Blue Willow Story Plates

    ERIC Educational Resources Information Center

    Fontes, Kris

    2009-01-01

    In the December 1997 issue of "SchoolArts" is a lesson titled "Blue Willow Story Plates" by Susan Striker. In this article, the author shares how she used this lesson with her middle-school students many times over the years. Here, she describes a Blue Willow plate painting project that her students made.

  19. Turbine vane plate assembly

    DOEpatents

    Schiavo Jr., Anthony L.

    2006-01-10

    A turbine vane assembly includes a turbine vane having first and second shrouds with an elongated airfoil extending between. Each end of the airfoil transitions into a shroud at a respective junction. Each of the shrouds has a plurality of cooling passages, and the airfoil has a plurality of cooling passages extending between the first and second shrouds. A substantially flat inner plate and an outer plate are coupled to each of the first and second shrouds so as to form inner and outer plenums. Each inner plenum is defined between at least the junction and the substantially flat inner plate; each outer plenum is defined between at least the substantially flat inner plate and the outer plate. Each inner plenum is in fluid communication with a respective outer plenum through at least one of the cooling passages in the respective shroud.

  20. Syncytial-Type Cell Plates

    PubMed Central

    Otegui, Marisa; Staehelin, L. Andrew

    2000-01-01

    Cell wall formation in the syncytial endosperm of Arabidopsis was studied by using high-pressure-frozen/freeze-substituted developing seeds and immunocytochemical techniques. The endosperm cellularization process begins at the late globular embryo stage with the synchronous organization of small clusters of oppositely oriented microtubules (∼10 microtubules in each set) into phragmoplast-like structures termed mini-phragmoplasts between both sister and nonsister nuclei. These mini-phragmoplasts produce a novel kind of cell plate, the syncytial-type cell plate, from Golgi-derived vesicles ∼63 nm in diameter, which fuse by way of hourglass-shaped intermediates into wide (∼45 nm in diameter) tubules. These wide tubules quickly become coated and surrounded by a ribosome-excluding matrix; as they grow, they branch and fuse with each other to form wide tubular networks. The mini-phragmoplasts formed between a given pair of nuclei produce aligned tubular networks that grow centrifugally until they merge into a coherent wide tubular network with the mini-phragmoplasts positioned along the network margins. The individual wide tubular networks expand laterally until they meet and eventually fuse with each other at the sites of the future cell corners. Transformation of the wide tubular networks into noncoated, thin (∼27 nm in diameter) tubular networks begins at multiple sites and coincides with the appearance of clathrin-coated budding structures. After fusion with the syncytial cell wall, the thin tubular networks are converted into fenestrated sheets and cell walls. Immunolabeling experiments show that the cell plates and cell walls of the endosperm differ from those of the embryo and maternal tissue in two features: their xyloglucans lack terminal fucose residues on the side chain, and callose persists in the cell walls after the cell plates fuse with the parental plasma membrane. The lack of terminal fucose residues on xyloglucans suggests that these cell wall

  1. Effects of meat juice on biofilm formation of Campylobacter and Salmonella.

    PubMed

    Li, Jiaqi; Feng, Jinsong; Ma, Lina; de la Fuente Núñez, César; Gölz, Greta; Lu, Xiaonan

    2017-07-17

    Campylobacter and Salmonella are leading causes of foodborne illnesses worldwide, vastly harboured by raw meat as their common food reservoir. Both microbes are prevalent in meat processing environments in the form of biofilms that contribute to cross-contamination and foodborne infection. This study applied raw meat juice (chicken juice and pork juice) as a minimally processed food model to study its effects on bacterial biofilm formation. Meat juice was collected during the freeze-thaw process of raw meat and sterilized by filtration. In 96-well polystyrene plates and glass chambers, supplementation of over 25% meat juice (v/v) in laboratory media led to an increase in biofilm formation of Campylobacter and Salmonella. During the initial attachment stage of biofilm development, more bacterial cells were present on surfaces treated with meat juice residues compared to control surfaces. Meat juice particulates on abiotic surfaces facilitated biofilm formation of Campylobacter and Salmonella under both static and flow conditions, with the latter being assessed using a microfluidic platform. Further, the deficiency in biofilm formation of selected Campylobacter and Salmonella mutant strains was restored in the presence of meat juice particulates. These results suggested that meat juice residues on the abiotic surfaces might act as a surface conditioner to support initial attachment and biofilm formation of Campylobacter and Salmonella. This study sheds light on a possible survival mechanism of Campylobacter and Salmonella in meat processing environments, and indicates that thorough cleaning of meat residues during meat production and handling is critical to reduce the bacterial load of Campylobacter and Salmonella. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Earthquakes and plate tectonics.

    USGS Publications Warehouse

    Spall, H.

    1982-01-01

    Earthquakes occur at the following three kinds of plate boundary: ocean ridges where the plates are pulled apart, margins where the plates scrape past one another, and margins where one plate is thrust under the other. Thus, we can predict the general regions on the earth's surface where we can expect large earthquakes in the future. We know that each year about 140 earthquakes of magnitude 6 or greater will occur within this area which is 10% of the earth's surface. But on a worldwide basis we cannot say with much accuracy when these events will occur. The reason is that the processes in plate tectonics have been going on for millions of years. Averaged over this interval, plate motions amount to several mm per year. But at any instant in geologic time, for example the year 1982, we do not know, exactly where we are in the worldwide cycle of strain build-up and strain release. Only by monitoring the stress and strain in small areas, for instance, the San Andreas fault, in great detail can we hope to predict when renewed activity in that part of the plate tectonics arena is likely to take place. -from Author

  3. Silver plating ensures reliable diffusion bonding of dissimilar metals

    NASA Technical Reports Server (NTRS)

    1967-01-01

    Dissimilar metals are reliably joined by diffusion bonding when the surfaces are electroplated with silver. The process involves cleaning and etching, anodization, silver striking, and silver plating with a conventional plating bath. It minimizes the formation of detrimental intermetallic phases and provides greater tolerance of processing parameters.

  4. Pixelated neutron image plates

    NASA Astrophysics Data System (ADS)

    Schlapp, M.; Conrad, H.; von Seggern, H.

    2004-09-01

    Neutron image plates (NIPs) have found widespread application as neutron detectors for single-crystal and powder diffraction, small-angle scattering and tomography. After neutron exposure, the image plate can be read out by scanning with a laser. Commercially available NIPs consist of a powder mixture of BaFBr : Eu2+ and Gd2O3 dispersed in a polymer matrix and supported by a flexible polymer sheet. Since BaFBr : Eu2+ is an excellent x-ray storage phosphor, these NIPs are particularly sensitive to ggr-radiation, which is always present as a background radiation in neutron experiments. In this work we present results on NIPs consisting of KCl : Eu2+ and LiF that were fabricated into ceramic image plates in which the alkali halides act as a self-supporting matrix without the necessity for using a polymeric binder. An advantage of this type of NIP is the significantly reduced ggr-sensitivity. However, the much lower neutron absorption cross section of LiF compared with Gd2O3 demands a thicker image plate for obtaining comparable neutron absorption. The greater thickness of the NIP inevitably leads to a loss in spatial resolution of the image plate. However, this reduction in resolution can be restricted by a novel image plate concept in which a ceramic structure with square cells (referred to as a 'honeycomb') is embedded in the NIP, resulting in a pixelated image plate. In such a NIP the read-out light is confined to the particular illuminated pixel, decoupling the spatial resolution from the optical properties of the image plate material and morphology. In this work, a comparison of experimentally determined and simulated spatial resolutions of pixelated and unstructured image plates for a fixed read-out laser intensity is presented, as well as simulations of the properties of these NIPs at higher laser powers.

  5. Plate tectonics, habitability and life

    NASA Astrophysics Data System (ADS)

    Spohn, Tilman; Breuer, Doris

    2016-04-01

    The role of plate tectonics in defining habitability of terrestrial planets is being increasingly discussed (e.g., Elkins-Tanton, 2015). Plate tectonics is a significantly evolved concept with a large variety of aspects. In the present context, cycling of material between near surface and mantle reservoirs is most important. But increased heat transport through mixing of cold lithosphere with the deep interior and formation of continental crust may also matter. An alternative mechanism of material cycling between these reservoirs is hot-spot volcanism combined with crust delamination. Hot-spot volcanism will transport volatiles to the atmosphere while delamination will mix crust, possibly altered by sedimentation and chemical reactions, with the mantle. The mechanism works as long as the stagnant lithosphere plate has not grown thicker than the crust and as long as volcanic material is added onto the crust. Thermal evolution studies suggest that the mechanism could work for the first 1-2 Ga of planetary evolution. The efficiency of the mechanism is limited by the ratio of extrusive to intrusive volcanism, which is thought to be less than 0.25. Plate tectonics would certainly have an advantage by working even for more evolved planets. A simple, most-used concept of habitability requires the thermodynamic stability of liquid water on the surface of a planet. Cycling of CO2between the atmosphere, oceans and interior through subduction and surface volcanism is an important element of the carbonate-silicate cycle, a thermostat feedback cycle that will keep the atmosphere from entering into a runaway greenhouse. Calculations for a model Earth lacking plate tectonics but degassing CO2, N, and H2O to form a surface ocean and a secondary atmosphere (Tosi et al, 2016) suggest that liquid water can be maintained on the surface for 4.5Ga. The model planet would then qualify as habitable. It is conceivable that the CO2 buffering capability of its ocean together with silicate

  6. Intra Plate Stresses Using Finite Element Modelling

    NASA Astrophysics Data System (ADS)

    Jayalakshmi, S.; Raghukanth, S. T. G.

    2016-10-01

    One of the most challenging problems in the estimation of seismic hazard is the ability to quantify seismic activity. Empirical models based on the available earthquake catalogue are often used to obtain activity of source regions. The major limitation with this approach is the lack of sufficient data near a specified source. The non-availability of data poses difficulties in obtaining distribution of earthquakes with large return periods. Such events recur over geological time scales during which tectonic processes, including mantle convection, formation of faults and new plate boundaries, are likely to take place. The availability of geometries of plate boundaries, plate driving forces, lithospheric stress field and GPS measurements has provided numerous insights on the mechanics of tectonic plates. In this article, a 2D finite element model of Indo-Australian plate is developed with the focus of representing seismic activity in India. The effect of large scale geological features including sedimentary basins, fold belts and cratons on the stress field in India is explored in this study. In order to address long term behaviour, the orientation of stress field and tectonic faults of the present Indo- Australian plate are compared with a reconstructed stress field from the early Miocene (20 Ma).

  7. What's On Your Plate?

    MedlinePlus

    ... Table of Contents What's On Your Plate? Smart Food Choices for Healthy Aging www.nia.nih.gov/health/ ... calories in" and "calories out," and making good food choices as you age. Shopping Tips See how planning ...

  8. Feynman's wobbling plate

    NASA Astrophysics Data System (ADS)

    Tuleja, Slavomir; Gazovic, Boris; Tomori, Alexander; Hanc, Jozef

    2007-03-01

    In the book Surely You Are Joking, Mr. Feynman! Richard Feynman tells a story of a Cornell cafeteria plate being tossed into the air. As the plate spun, it wobbled. Feynman noticed a relation between the two motions. He solved the motion of the plate by using the Lagrangian approach. This solution didn't satisfy him. He wanted to understand the motion of the plate by analyzing the motion of its individual particles and the forces acting on them. He was successful, but he didn't tell us how he did it. We provide an elementary explanation for the two-to-one ratio of wobble to spin frequencies, based on an analysis of the motion of the particles and the forces acting on them. We also demonstrate the power of numerical simulation and computer animation to provide insight into a physical phenomenon and guidance on how to do the analysis.

  9. Reduction of astrometric plates

    NASA Technical Reports Server (NTRS)

    Stock, J.

    1984-01-01

    A rapid and accurate method for the reduction of comet or asteroid plates is described. Projection equations, scale length correction, rotation of coordinates, linearization, the search for additional reference stars, and the final solution are examined.

  10. Violin plate modes.

    PubMed

    Gough, Colin

    2015-01-01

    As the first step toward developing a generic model for the acoustically radiating vibrational modes of the violin and related instruments, the modes of both freely supported and edge-constrained top and back plates have been investigated as functions of shape, arching height, elastic anisotropy, the f-holes and associated island area, thickness graduations, and the additional boundary constraints of the ribs, soundpost, and bass-bar present in the assembled instrument. Comsol shell structure finite element software has been used as a quasi-experimental tool, with physical and geometric properties varied smoothly, often over several orders of magnitude, allowing the development of the plate modes to be followed continuously from those of an initially square plate to those of doubly-arched, guitar-shaped, orthotropic plates and their dependence on all the above factors.

  11. Reduction of astrometric plates

    NASA Technical Reports Server (NTRS)

    Stock, J.

    1984-01-01

    A rapid and accurate method for the reduction of comet or asteroid plates is described. Projection equations, scale length correction, rotation of coordinates, linearization, the search for additional reference stars, and the final solution are examined.

  12. Tectonic Plate Movement.

    ERIC Educational Resources Information Center

    Landalf, Helen

    1998-01-01

    Presents an activity that employs movement to enable students to understand concepts related to plate tectonics. Argues that movement brings topics to life in a concrete way and helps children retain knowledge. (DDR)

  13. Plate tectonics: Metamorphic myth

    NASA Astrophysics Data System (ADS)

    Korenaga, Jun

    2016-01-01

    Clear evidence for subduction-induced metamorphism, and thus the operation of plate tectonics on the ancient Earth has been lacking. Theoretical calculations indicate that we may have been looking for something that cannot exist.

  14. Tectonic Plate Movement.

    ERIC Educational Resources Information Center

    Landalf, Helen

    1998-01-01

    Presents an activity that employs movement to enable students to understand concepts related to plate tectonics. Argues that movement brings topics to life in a concrete way and helps children retain knowledge. (DDR)

  15. Designing Assemblies Of Plates

    NASA Technical Reports Server (NTRS)

    Williams, F. W.; Kennedy, D.; Butler, R.; Aston, G.; Anderson, M. S.

    1992-01-01

    VICONOPT calculates vibrations and instabilities of assemblies of prismatic plates. Designed for efficient, accurate analysis of buckling and vibration, and for optimum design of panels of composite materials. Written in FORTRAN 77.

  16. Flat plate solar oven

    SciTech Connect

    Parikh, M.

    1981-01-01

    The construction of an Indian Rs. 186 (US $20.33) flat-plate solar oven is described. Detailed drawings are provided and relevant information on cooking times and temperature for different foods is given.

  17. Positive battery plate

    NASA Technical Reports Server (NTRS)

    Rowlette, John R. (Inventor)

    1985-01-01

    The power characteristics of a lead acid battery are improved by incorporating a dispersion of 1 to 10% by weight of a thermodynamically stable conductivity additive, such as conductive tin oxide coated glass fibers (34) of filamentary glass wool (42) in the positive active layer (32) carried on the grid (30) of the positive plate (16). Positive plate potential must be kept high enough to prevent reduction of the tin oxide to tin by utilizing an oversized, precharged positive paste.

  18. Fractal multifiber microchannel plates

    NASA Technical Reports Server (NTRS)

    Cook, Lee M.; Feller, W. B.; Kenter, Almus T.; Chappell, Jon H.

    1992-01-01

    The construction and performance of microchannel plates (MCPs) made using fractal tiling mehtods are reviewed. MCPs with 40 mm active areas having near-perfect channel ordering were produced. These plates demonstrated electrical performance characteristics equivalent to conventionally constructed MCPs. These apparently are the first MCPs which have a sufficiently high degree of order to permit single channel addressability. Potential applications for these devices and the prospects for further development are discussed.

  19. Polymicrobial biofilm formation by Candida albicans, Actinomyces naeslundii, and Streptococcus mutans is Candida albicans strain and medium dependent.

    PubMed

    Arzmi, Mohd Hafiz; Alnuaimi, Ali D; Dashper, Stuart; Cirillo, Nicola; Reynolds, Eric C; McCullough, Michael

    2016-11-01

    Oral biofilms comprise of extracellular polysaccharides and polymicrobial microorganisms. The objective of this study was to determine the effect of polymicrobial interactions of Candida albicans, Actinomyces naeslundii, and Streptococcus mutans on biofilm formation with the hypotheses that biofilm biomass and metabolic activity are both C. albicans strain and growth medium dependent. To study monospecific biofilms, C. albicans, A. naeslundii, and S. mutans were inoculated into artificial saliva medium (ASM) and RPMI-1640 in separate vials, whereas to study polymicrobial biofilm formation, the inoculum containing microorganisms was prepared in the same vial prior inoculation into a 96-well plate followed by 72 hours incubation. Finally, biofilm biomass and metabolic activity were measured using crystal violet and XTT assays, respectively. Our results showed variability of monospecies and polymicrobial biofilm biomass between C. albicans strains and growth medium. Based on cut-offs, out of 32, seven RPMI-grown biofilms had high biofilm biomass (HBB), whereas, in ASM-grown biofilms, 14 out of 32 were HBB. Of the 32 biofilms grown in RPMI-1640, 21 were high metabolic activity (HMA), whereas in ASM, there was no biofilm had HMA. Significant differences were observed between ASM and RPMI-grown biofilms with respect to metabolic activity (P <01). In conclusion, biofilm biomass and metabolic activity were both C. albicans strain and growth medium dependent. © The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Formation of the giant Chalukou porphyry Mo deposit in northern Great Xing'an Range, NE China: Partial melting of the juvenile lower crust in intra-plate extensional environment

    NASA Astrophysics Data System (ADS)

    Li, Zhen-Zhen; Qin, Ke-Zhang; Li, Guang-Ming; Ishihara, Shunso; Jin, Lu-Ying; Song, Guo-Xue; Meng, Zhao-Jun

    2014-08-01

    The Chalukou porphyry Mo deposit (2.46 Mt @ 0.087% Mo), located in the northern Great Xing'an Range, NE China, is the largest Mo deposit discovered in China so far. The host rocks consist of aplite porphyry, granite porphyry and quartz porphyry, and are intruded into Lower Ordovician intermediate-felsic volcanic-sedimentary rocks and pre-ore monzogranite and are cut by post-ore feldspar porphyry, diorite porphyry and quartz monzonite porphyry. Here, we present the zircon U-Pb ages, whole-rock geochemistry, Sr-Nd isotopic and zircon Hf isotopic data for the pre-ore, syn-ore and post-ore intrusive rocks. The Chalukou ore-forming porphyries intruded during 147-148 Ma and have high-silica, alkali-rich, metaluminous to slightly peraluminous compositions and are oxidized. They are enriched in large ion lithophile elements (e.g. K, Rb, U and Th), light REE and depleted in high-field strength elements (e.g. Nb, P and Ti). Depletions in Eu, Ba, Sr, Nb, Ta, P and Ti suggest that they have experienced strong fractional crystallization of plagioclase, biotite, hornblende and accessory minerals. The pre-ore monzogranite (~ 172 Ma) also belongs to the high-K calc-alkaline series. Highly fractionated REE patterns ((La/Yb) N = 19.6-21.7), high values of Sr/Y (54-69) and La/Yb (29-32), are adakite-like geochemical features. The post-ore rocks (~ 141-128 Ma) have similar geochemical characteristics with ore-forming porphyries except that quartz monzonite porphyry shows no Ba-Sr negative anomaly. All intrusive rocks have relative low initial 87Sr/86Sr (0.705413-0.707889) and εNd (t) values (- 1.28 to + 0.92), positive εHf (t) values (+ 2.4 to + 10.1) and young two-stage Nd and Hf model ages (TDM2 (Nd) = 863-977 Ma, TDM2 (Hf) = 552-976 Ma). These geochemical and isotopic data are interpreted to demonstrate that the ore-forming porphyries formed by partial melting of the juvenile lower crust caused by underplating of mafic magmas in an intra-plate extensional setting. The pre

  1. Inhibiting effects of Streptococcus salivarius on competence-stimulating peptide-dependent biofilm formation by Streptococcus mutans.

    PubMed

    Tamura, S; Yonezawa, H; Motegi, M; Nakao, R; Yoneda, S; Watanabe, H; Yamazaki, T; Senpuku, H

    2009-04-01

    The effects of Streptococcus salivarius on the competence-stimulating peptide (CSP)-dependent biofilm formation by Streptococcus mutans were investigated. Biofilms were grown on 96-well microtiter plates coated with salivary components in tryptic soy broth without dextrose supplemented with 0.25% sucrose. Biofilm formations were stained using safranin and quantification of stained biofilms was performed by measuring absorbance at 492 nm. S. mutans formed substantial biofilms, whereas biofilms of S. salivarius were formed poorly in the medium conditions used. Furthermore, in combination cultures, S. salivarius strongly inhibited biofilm formation when cultured with S. mutans. This inhibition occurred in the early phase of biofilm formation and was dependent on inactivation of the CSP of S. mutans, which is associated with competence, biofilm formation, and antimicrobial activity of the bacterium, and is induced by expression of the comC gene. Comparisons between the S. mutans clinical strains FSC-3 and FSC-3DeltaglrA in separate dual-species cultures with S. salivarius indicated that the presence of the bacitracin transport ATP-binding protein gene glrA caused susceptibility to inhibition of S. mutans biofilm formation by S. salivarius, and was also associated with the regulation of CSP production by com gene-dependent quorum sensing systems. It is considered that regulation of CSP by glrA in S. mutans and CSP inactivation by S. salivarius are important functions for cell-to-cell communication between biofilm bacteria and oral streptococci such as S. salivarius. Our results provide useful information for understanding the ecosystem of oral streptococcal biofilms, as well as the competition between and coexistence of multiple species in the oral cavity.

  2. Distinct modes of floor plate induction in the chick embryo.

    PubMed

    Patten, Iain; Kulesa, Paul; Shen, Michael M; Fraser, Scott; Placzek, Marysia

    2003-10-01

    To begin to reconcile models of floor plate formation in the vertebrate neural tube, we have performed experiments aimed at understanding the development of the early floor plate in the chick embryo. Using real-time analyses of cell behaviour, we provide evidence that the principal contributor to the early neural midline, the future anterior floor plate, exists as a separate population of floor plate precursor cells in the epiblast of the gastrula stage embryo, and does not share a lineage with axial mesoderm. Analysis of the tissue interactions associated with differentiation of these cells to a floor plate fate reveals a role for the nascent prechordal mesoderm, indicating that more than one inductive event is associated with floor plate formation along the length of the neuraxis. We show that Nr1, a chick nodal homologue, is expressed in the nascent prechordal mesoderm and we provide evidence that Nodal signalling can cooperate with Shh to induce the epiblast precursors to a floor-plate fate. These results indicate that a shared lineage with axial mesoderm cells is not a pre-requisite for floor plate differentiation and suggest parallels between the development of the floor plate in amniote and anamniote embryos.

  3. Calcium increases Xylella fastidiosa surface attachment, biofilm formation, and twitching motility.

    PubMed

    Cruz, Luisa F; Cobine, Paul A; De La Fuente, Leonardo

    2012-03-01

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

  4. Caribbean plate interactions

    SciTech Connect

    Ball, M. )

    1993-02-01

    Vector analysis of plate motions, derived from studies of Atlantic magnetic lineations and fracture zone trends, indicates the following relative movements between the Caribbean, North American, and South American Plates. (1) During Early Jurassic to Early Cretaceous, the North American Plate moved 1900 km westward and 900 km northward relative to the South American Plate. A broad zone including the Caribbean region, i.e., the zone between the North and South America Plates, was a site of left-lateral shear and north-south extension. (2) During Early Cretaceous to Late Cretaceous, the North American Mate moved an additional 1200 km westward relative to South America across this zone. (3) During Late Cretaceous to the end of the Eocene, the North American Plate moved 200 km westward and 400 km northward relative to the South American Plate. (4) From the end of the Eocene to near the end of the Miocene, North America converged on South America some 200 km and moved 100 km eastward relative to it. Through the Mesozoic and earliest Tertiary history of the Caribbean, the region was a shear zone within which left-lateral displacement exceeded 3000 km and north-south extension exceeded 1300 km. In regard to time, 80% of the history of the Caribbean region is one of north-south extension and left-lateral shear. In terms of space, 97% of the shear is left-lateral and the ratio of divergence versus convergence is 7 to 1. Thus, characterizing the Caribbean region, and the Atlantic to its east, as a zone of north-south extension and left-lateral shear, is a fair generalization.

  5. Effects of the microstructure of twin roll cast and hot rolled plates on the surface quality of presensitized plates

    NASA Astrophysics Data System (ADS)

    Zhu, Yuan-Zhi; Zhang, Ya-Feng; Zhao, Chao-Qi; Zhou, Feng

    2014-09-01

    The effect of the microstructure of plates fabricated both in the traditional process, involving casting, hot rolling and cold rolling (HR), and in the novel twin roll casting + cold rolling (TRC) process on the surface quality of presensitized (PS) plates was analyzed by optical microscopy (OM), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDXS). The formation of pores on the surface of the electrolyzed HR plate could be attributed to the presence of approximately 1-μm-sized large Al-Fe precipitates in the HR plate compared to the smaller precipitates in the TRC plate. Moreover, residual graphite lubricants used during the TRC process were entrapped on the surface of the TRC plate during the subsequent rolling process. The entrapped pollutants tended to further deteriorate the formation of pores on the surface of the TRC plate, and no residual carbon was detected on the surface of the HR plate. Furthermore, the surface quality of the TRC plate can be improved by surface cleaning before the cold rolling process, which could dramatically lower the residual graphite on the surface.

  6. When Did Plate Tectonics Begin

    NASA Astrophysics Data System (ADS)

    Brown, M.

    2015-12-01

    Present-day plate tectonics on Earth is characterized by asymmetric (one-sided) subduction, but how do we recognize the imprint of subduction in the geologic record? How do we weigh global (commonly younger) vs local (commonly older) datasets or distinguish initiation from episodic from continuous subduction? How reliable are data gaps? Characteristics of the Paleozoic record of subduction include calc-alkaline magmatism, blueschist/UHP metamorphism and collisional orogenesis, and ophiolites as representatives of former ocean lithosphere. Are these characteristic rocks preserved in Proterozoic, Archean and Hadean crust? Does a hotter mantle, higher heat production and weaker lithosphere modify or eliminate these features? What preceded subduction and how do we recognize that regime? Are rock associations or geochemical fingerprints reliable? Does reworking and overprinting modify geochemical fingerprints? Proposals for the start of plate tectonics have been based on: persistence of isotope anomalies/fractionated chemical domains in the mantle; changes in chemistry of magmatic rocks, rates of crustal growth vs reworking, and sites of growth; the metamorphic record, particularly the first appearance of contrasting thermal gradients or eclogite (including evidence from mineral inclusions in diamonds) or UHP metamorphic rocks; stabilization of cratonic lithosphere and formation of supercratons, and the beginning of the Proterozoic supercontinent cycle; the end of the flat Earth, emergence of continents, development of significant topography, changes in the style of orogeny and the rise in atmospheric oxygen; and, the appearance of passive margins and changes in the style of sedimentation. Estimates of the timing have varied from the Hadean to Neoproterozoic. I will summarize evidence for a growing consensus that the late Mesoarchean to early Paleoproterozoic was a 700 Myr long period of transition to continuous (?) subduction and global (?) mobile-lid plate tectonics.

  7. Cadmium plating replacements

    NASA Technical Reports Server (NTRS)

    Nelson, Mary J.; Groshart, Earl C.

    1995-01-01

    The Boeing Company has been searching for replacements to cadmium plate. Two alloy plating systems seem close to meeting the needs of a cadmium replacement. The two alloys, zinc-nickel and tin-zinc are from alloy plating baths; both baths are neutral pH. The alloys meet the requirements for salt fog corrosion resistance, and both alloys excel as a paint base. Currently, tests are being performed on standard fasteners to compare zinc-nickel and tin-zinc on threaded hardware where cadmium is heavily used. The Hydrogen embrittlement propensity of the zinc-nickel bath has been tested, and just beginning for the tin-zinc bath. Another area of interest is the electrical properties on aluminum for tin-zinc and will be discussed. The zinc-nickel alloy plating bath is in production in Boeing Commercial Airplane Group for non-critical low strength steels. The outlook is promising that these two coatings will help The Boeing Company significantly reduce its dependence on cadmium plating.

  8. Cadmium plating replacements

    SciTech Connect

    Nelson, M.J.; Groshart, E.C.

    1995-03-01

    The Boeing Company has been searching for replacements to cadmium plate. Two alloy plating systems seem close to meeting the needs of a cadmium replacement. The two alloys, zinc-nickel and tin-zinc are from alloy plating baths; both baths are neutral pH. The alloys meet the requirements for salt fog corrosion resistance, and both alloys excel as a paint base. Currently, tests are being performed on standard fasteners to compare zinc-nickel and tin-zinc on threaded hardware where cadmium is heavily used. The Hydrogen embrittlement propensity of the zinc-nickel bath has been tested, and just beginning for the tin-zinc bath. Another area of interest is the electrical properties on aluminum for tin-zinc and will be discussed. The zinc-nickel alloy plating bath is in production in Boeing Commercial Airplane Group for non-critical low strength steels. The outlook is promising that these two coatings will help The Boeing Company significantly reduce its dependence on cadmium plating.

  9. Polar Plate Theory for Orthogonal Anisotropy

    NASA Technical Reports Server (NTRS)

    Bailey, Michelle D.; Bower, Mark V.

    2000-01-01

    evaluate out-of-plane displacements for given load conditions in design of composite Structures. Without the formation of these unique plate equations, design of such structures is not a precise engineering accomplishment due to the lack of precise design tools. Such structures are over designed to compensate and costly tests need to be performed after production to validate safety. Though impossible to present the mathematical solution within the limits of this paper, the application of the newly derived plate equation in its orthotropic form for a balanced symmetric laminate compared favorably with test results and surpassed the results of standard numerical methods. Within the limitations of the problem foundation the predictive model provides a mechanism for establishing out-of-plane deflection levels for a circular composite plate. Future work in this area should include incorporating the circular composite plate equations in Finite Element Models. The new anisotropic polar plate equations call be utilized as a design tool so that the design more accurately meets the requirements, thus taking full advantage of the weight savings of composites. These accomplishments have not previously been presented to the engineering community nor ever proposed to the IAF. Should the proposed paper be accepted for presentation, attendance is assured.

  10. Bipolar battery plate

    NASA Technical Reports Server (NTRS)

    Rowlette, John J. (Inventor)

    1987-01-01

    A liquid-impermeable plate (10) having through-plate conductivity with essentially zero resistance comprises an insulator sheet (12) having a series of spaced perforations (14) each of which contains a metal element (16) sealingly received into the perforation (14). A low-cost plate can readily be manufactured by punching a thermoplastic sheet (40) such as polypropylene with a punching tool (52), filling the apertures with led spheres (63) having a diameter smaller than the holes (50) but larger than the thickness of the sheet, sweeping excess spheres (62) off the sheet with a doctor blade (60) and then pressing a heated platen (74) onto the sheet to swage the spheres into a cylindrical shape and melt the surrounding resin to form a liquid-impermeable collar (4) sealing the metal into the sheet.

  11. Plate-mantle coupling from post-Pangea plate kinematics

    NASA Astrophysics Data System (ADS)

    Zahirovic, Sabin; Dietmar Müller, R.; Seton, Maria; Flament, Nicolas

    2015-04-01

    Convection in the Earth's mantle that involves plates at the surfaces gives rise to plate velocities that vary through time and depend on the balance of plate boundary forces, with the present-day providing a snapshot of this ongoing process. However, present-day plate velocities do not capture plate behaviour over geologically representative timeframes and thus cannot be used to evaluate factors limiting plate velocities. Previous studies investigated the effects of continental keels on plate speeds by either using the present-day snapshot or a limited number of reconstructed plate configurations, often leading to conflicting results. For example, an early assumption was that continental keels (especially cratons) were unlikely to impede fast plate motions because India's velocity approached ~20 cm/yr in the Eocene prior to the collision with Eurasia. We employ a modern plate reconstruction approach with evolving global topological plate boundaries for the post-Pangea timeframe (since 200 Ma) to evaluate factors controlling plate velocities. Plate boundary configurations and plate velocities are extracted from the open-source and cross-platform plate reconstruction package GPlates (www.gplates.org) at 1 Myr intervals. For each plate, at each timestep, the area of continental and cratonic lithosphere is calculated to evaluate the effect on plate velocities. Our results support that oceanic plates tend to be 2-3 times faster than plates with large portion of continental plate area, consistent with predictions of numerical models of mantle convection. The fastest plates (~8.5 cm/yr RMS) are dominated by oceanic plate area and high subducting portion of plate perimeter, while the slowest plates (~2.6-2.8 cm/yr RMS) are dominated by continental plate area and bounded by transforms and mid-oceanic ridge segments. Importantly, increasing cratonic fractions (both Proterozoic and Archean lithosphere) significantly impede plate velocities, suggesting that deep continental

  12. Reduced Plating Ignitron

    NASA Technical Reports Server (NTRS)

    Polzin, Kurt A (Inventor); Pearson, J Boise (Inventor)

    2014-01-01

    An ignitron apparatus has an airtight tubular housing having a first sealed end and a second sealed end. An anode is connected at the first sealed end, projecting into the housing, and a recess at the second sealed and forms a well which contains a quantity of liquid gallium or gallium alloy making up the cathode. An ignitor projects through the liquid metal and into the housing. The inner surface of the housing includes at least one plating-reduction structure to prevent electrical shorting of the apparatus caused by plating of the liquid metal.

  13. License plate detection algorithm

    NASA Astrophysics Data System (ADS)

    Broitman, Michael; Klopovsky, Yuri; Silinskis, Normunds

    2013-12-01

    A novel algorithm for vehicle license plates localization is proposed. The algorithm is based on pixel intensity transition gradient analysis. Near to 2500 natural-scene gray-level vehicle images of different backgrounds and ambient illumination was tested. The best set of algorithm's parameters produces detection rate up to 0.94. Taking into account abnormal camera location during our tests and therefore geometrical distortion and troubles from trees this result could be considered as passable. Correlation between source data, such as license Plate dimensions and texture, cameras location and others, and parameters of algorithm were also defined.

  14. Plating under reduced pressure

    SciTech Connect

    Dini, J.W.; Beat, T.G.; Cowden, W.C. ); Ryan, L.E.; Hewitt, W.B. )

    1992-06-01

    Plating under reduced pressure was evaluated for both electroless nickel and electrodeposited copper systems. The objective was to reduce pitting of these coatings thereby further enhancing their usage for diamond turning applications. Cursory experiments with electroless nickel showed reduced porosity when deposition was done at around 500 torr. Detailed experiments with electrodeposited copper at around 100 torr provided similar results. Scanning tunneling microscopy was effectively used to show the improvement in the copper deposits plated under reduced pressure. Benefits included reduced surface roughness and finer and denser grain structure.

  15. NICKEL PLATING PROCESS

    DOEpatents

    Hoover, T.B.; Zava, T.E.

    1959-05-12

    A simplified process is presented for plating nickel by the vapor decomposition of nickel carbonyl. In a preferred form of the invention a solid surface is nickel plated by subjecting the surface to contact with a mixture containing by volume approximately 20% nickel carbonyl vapor, 2% hydrogen sulfide and .l% water vapor or 1% oxygen and the remainder carbon dioxide at room temperature until the desired thickness of nickel is obtained. The advantage of this composition over others is that the normally explosive nickel carbonyl is greatly stabilized.

  16. PCM Composite Cold Plate.

    DTIC Science & Technology

    1996-04-01

    packaging design for phase change materials (PCMs) used in thermal management. To control expansion stress, conventional PCM heat sinks have strong encapsulation that weighs more than the PCM they contain. The objective of this work is to develop PCM heat sinks configured as thin sandwich plates, in which the mass of the encapsulation is a small fraction of the PCM mass. The design is based on a lightweight carbon fiber core that has suitable mechanical, thermal, and capillary properties. PCM composite plates were fabricated with dimensions 15x15x0.6 cm3 and with a

  17. Comparison of the effect on bone healing process of different implants used in minimally invasive plate osteosynthesis: limited contact dynamic compression plate versus locking compression plate

    PubMed Central

    Xue, Zichao; Xu, Haitao; Ding, Haoliang; Qin, Hui; An, Zhiquan

    2016-01-01

    Minimally invasive plate osteosynthesis (MIPO) has been widely accepted because of its satisfactory clinical outcomes. However, the implant construct that works best for MIPO remains controversial. Different plate designs result in different influence mechanisms to blood flow. In this study, we created ulnar fractures in 42 beagle dogs and fixed the fractures using MIPO. The dogs were randomly divided into two groups and were fixed with a limited contact dynamic compression plate (LC-DCP) or a locking compression plate (LCP). Our study showed that with MIPO, there was no significant difference between the LCP and the LC-DCP in terms of fracture fixation, bone formation, or mineralization. Combined with the previous literature, we inferred that the healing process is affected by the quality of fracture reduction more than plate selection. PMID:27885262

  18. 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.

  19. Elastocapillary coalescence of plates and pillars

    PubMed Central

    Wei, Z.; Schneider, T. M.; Kim, J.; Kim, H.-Y.; Aizenberg, J.; Mahadevan, L.

    2015-01-01

    When a fluid-immersed array of supported plates or pillars is dried, evaporation leads to the formation of menisci on the tips of the plates or pillars that bring them together to form complex patterns. Building on prior experimental observations, we use a combination of theory and computation to understand the nature of this instability and its evolution in both the two- and three-dimensional setting of the problem. For the case of plates, we explicitly derive the interaction torques based on the relevant physical parameters associated with pillar deformation, contact-line pinning/depinning and fluid volume changes. A Bloch-wave analysis for our periodic mechanical system captures the window of volumes where the two-plate eigenvalue characterizes the onset of the coalescence instability. We then study the evolution of these binary clusters and their eventual elastic arrest using numerical simulations that account for evaporative dynamics coupled to capillary coalescence. This explains both the formation of hierarchical clusters and the sensitive dependence of the final structures on initial perturbations, as seen in our experiments. We then generalize our analysis to treat the problem of pillar collapse in three dimensions, where the fluid domain is completely connected and the interface is a minimal surface with the uniform mean curvature. Our theory and simulations capture the salient features of experimental observations in a range of different situations and may thus be useful in controlling the ensuing patterns. PMID:25792949

  20. Interaction of Vortex Ring with Cutting Plate

    NASA Astrophysics Data System (ADS)

    Musta, Mustafa

    2015-11-01

    The interaction of a vortex ring impinging on a thin cutting plate was made experimentally using Volumetric 3-component Velocitmetry (v3v) technique. The vortex rings were generated with piston-cylinder vortex ring generator using piston stroke-to-diameter ratios and Re at 2-3 and 1500 - 3000, respectively. The cutting of vortex rings below center line leads to the formation of secondary vortices on each side of the plate which is look like two vortex rings, and a third vortex ring propagates further downstream in the direction of the initial vortex ring, which is previously showed by flow visualization study of Weigand (1993) and called ``trifurcation''. Trifurcation is very sensitive to the initial Reynolds number and the position of the plate with respect to the vortex ring generator pipe. The present work seeks more detailed investigation on the trifurcation using V3V technique. Conditions for the formation of trifurcation is analyzed and compared with Weigand (1993). The formed secondary vortex rings and the propagation of initial vortex ring in the downstream of the plate are analyzed by calculating their circulation, energy and trajectories.

  1. Nuclear reactor alignment plate configuration

    SciTech Connect

    Altman, David A; Forsyth, David R; Smith, Richard E; Singleton, Norman R

    2014-01-28

    An alignment plate that is attached to a core barrel of a pressurized water reactor and fits within slots within a top plate of a lower core shroud and upper core plate to maintain lateral alignment of the reactor internals. The alignment plate is connected to the core barrel through two vertically-spaced dowel pins that extend from the outside surface of the core barrel through a reinforcement pad and into corresponding holes in the alignment plate. Additionally, threaded fasteners are inserted around the perimeter of the reinforcement pad and into the alignment plate to further secure the alignment plate to the core barrel. A fillet weld also is deposited around the perimeter of the reinforcement pad. To accomodate thermal growth between the alignment plate and the core barrel, a gap is left above, below and at both sides of one of the dowel pins in the alignment plate holes through with the dowel pins pass.

  2. 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. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Effect of cinnamon oil on icaA expression and biofilm formation by Staphylococcus epidermidis.

    PubMed

    Nuryastuti, Titik; van der Mei, Henny C; Busscher, Henk J; Iravati, Susi; Aman, Abu T; Krom, Bastiaan P

    2009-11-01

    Staphylococcus epidermidis is notorious for its biofilm formation on medical devices, and novel approaches to prevent and kill S. epidermidis biofilms are desired. In this study, the effect of cinnamon oil on planktonic and biofilm cultures of clinical S. epidermidis isolates was evaluated. Initially, susceptibility to cinnamon oil in planktonic cultures was compared to the commonly used antimicrobial agents chlorhexidine, triclosan, and gentamicin. The MIC of cinnamon oil, defined as the lowest concentration able to inhibit visible microbial growth, and the minimal bactericidal concentration, the lowest concentration required to kill 99.9% of the bacteria, were determined using the broth microdilution method and plating on agar. A checkerboard assay was used to evaluate the possible synergy between cinnamon oil and the other antimicrobial agents. The effect of cinnamon oil on biofilm growth was studied in 96-well plates and with confocal laser-scanning microscopy (CLSM). Biofilm susceptibility was determined using a metabolic 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Real-time PCR analysis was performed to determine the effect of sub-MIC concentrations of cinnamon oil on expression of the biofilm-related gene, icaA. Cinnamon oil showed antimicrobial activity against both planktonic and biofilm cultures of clinical S. epidermidis strains. There was only a small difference between planktonic and biofilm MICs, ranging from 0.5 to 1% and 1 to 2%, respectively. CLSM images indicated that cinnamon oil is able to detach and kill existing biofilms. Thus, cinnamon oil is an effective antimicrobial agent to combat S. epidermidis biofilms.

  4. The Plate Tectonics Project

    ERIC Educational Resources Information Center

    Hein, Annamae J.

    2011-01-01

    The Plate Tectonics Project is a multiday, inquiry-based unit that facilitates students as self-motivated learners. Reliable Web sites are offered to assist with lessons, and a summative rubric is used to facilitate the holistic nature of the project. After each topic (parts of the Earth, continental drift, etc.) is covered, the students will…

  5. The Plate Tectonics Project

    ERIC Educational Resources Information Center

    Hein, Annamae J.

    2011-01-01

    The Plate Tectonics Project is a multiday, inquiry-based unit that facilitates students as self-motivated learners. Reliable Web sites are offered to assist with lessons, and a summative rubric is used to facilitate the holistic nature of the project. After each topic (parts of the Earth, continental drift, etc.) is covered, the students will…

  6. INL HIP Plate Fabrication

    SciTech Connect

    B. H. Park; C. R. Clark; J. F. Jue

    2010-02-01

    This document outlines the process used to bond monolithic fuel plates by Hot Isostatic Pressing (HIP). This method was developed at Idaho National Laboratory (INL) for the Reduced Enrichment for Research and Test Reactors (RERTR) program. These foils have been used in a number of irradiation experiments in support of the United States Global Threat Reduction Initiative (GTRI) program.

  7. Unitary plate electrode

    NASA Technical Reports Server (NTRS)

    Rowlette, John J. (Inventor); Clough, Thomas J. (Inventor); Josefowicz, Jack Y. (Inventor); Sibert, John W. (Inventor)

    1985-01-01

    The unitary electrode (10) comprises a porous sheet (12) of fiberglass the strands (14) of which contain a coating (16) of conductive tin oxide. The lower portion of the sheet contains a layer (18) of resin and the upper layer (20) contains lead dioxide forming a positive active electrode on an electrolyte-impervious layer. The strands (14) form a continuous conduction path through both layers (16, 18). Tin oxide is prevented from reduction by coating the surface of the plate facing the negative electrode with a conductive, impervious layer resistant to reduction such as a thin film (130) of lead or graphite filled resin adhered to the plate with a layer (31) of conductive adhesive. The plate (10) can be formed by casting a molten resin from kettle (60) onto a sheet of glass wool (56) overlying a sheet of lead foil and then applying positive active paste from hopper (64) into the upper layer (68). The plate can also be formed by passing an assembly of a sheet ( 80) of resin, a sheet (86) of sintered glass and a sheet (90) of lead between the nip (92) of heated rollers (93, 95) and then filling lead oxide into the pores (116) of the upper layer (118).

  8. 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.

  9. 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.

  10. On the Enigmatic Birth of the Pacific Plate within the Panthalassa Ocean

    NASA Astrophysics Data System (ADS)

    Boschman, L.; Van Hinsbergen, D. J. J.

    2016-12-01

    The oceanic Pacific Plate started forming in Early Jurassic time within the vast Panthalassa Ocean that surrounded the supercontinent Pangea and contains the oldest lithosphere that can directly constrain the geodynamic history of the circum-Pangean Earth. Here, we show that the geometry of the oldest marine magnetic anomalies of the Pacific Plate attests of a unique plate kinematic event that sparked the plate's birth in virtually a point location, surrounded by the Izanagi, Farallon and Phoenix Plates. We reconstruct the unstable triple junction that caused the plate reorganization leading to the birth of the Pacific Plate and present a model of the plate tectonic configuration that preconditioned this event. We show that a stable, but migrating triple junction involving the gradual cessation of intra-oceanic Panthalassa subduction culminated in the formation of an unstable transform-transform-transform triple junction. The consequent plate boundary reorganization resulted in the formation of a stable triangular three-ridge system from which the nascent Pacific Plate expanded. We link the birth of the Pacific Plate to the regional termination of intra-Panthalassa subduction. Remnants thereof have been identified in the deep lower mantle of which the locations may provide paleolongitudinal control on the absolute location of the early Pacific Plate. Our results constitute an essential step in unraveling the plate tectonic evolution of `Thalassa Incognita' comprising the comprehensive Panthalassa Ocean surrounding Pangea.

  11. What Are Growth Plate Injuries?

    MedlinePlus

    ... for growth plate injuries are:  Falling down  Competitive sports (like football)  Recreational activities. Other reasons for growth plate injuries are:  Child abuse  Injury from extreme cold (for example, frostbite)  Radiation (used to treat ...

  12. Renewable liquid reflecting zone plate

    DOEpatents

    Toor, Arthur; Ryutov, Dmitri D.

    2003-12-09

    A renewable liquid reflecting zone plate. Electrodes are operatively connected to a dielectric liquid in a circular or other arrangement to produce a reflecting zone plate. A system for renewing the liquid uses a penetrable substrate.

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

    DTIC Science & Technology

    2015-05-13

    study and in vitro exposure experiments, Environ. Health Perspect. 114 (2006) 70–76. [12] A. Tsugita, M. Inouye, Purification of bacteriophage T4...Rennell, S.E. Bouvier, L.W. Hardy, A.R. Poteete, Systematicmutation of bacteriophage T4 lysozyme, J Mol. Biol. 222 (1991) 67–88. [18] L.W. Hardy, A.R...Poteete, Reexamination of the role of Asp20 in catalysis by bacteriophage T4 lysozyme, Biochemistry (Mosc.) 30 (1991) 9457–9463. [19] J.A. Knight

  14. Microchannel plate streak camera

    DOEpatents

    Wang, Ching L.

    1989-01-01

    An improved streak camera in which a microchannel plate electron multiplier is used in place of or in combination with the photocathode used in prior streak cameras. The improved streak camera is far more sensitive to photons (UV to gamma-rays) than the conventional x-ray streak camera which uses a photocathode. The improved streak camera offers gamma-ray detection with high temporal resolution. It also offers low-energy x-ray detection without attenuation inside the cathode. Using the microchannel plate in the improved camera has resulted in a time resolution of about 150 ps, and has provided a sensitivity sufficient for 1000 KeV x-rays.

  15. North American plate dynamics

    NASA Technical Reports Server (NTRS)

    Richardson, Randall M.; Reding, Lynn M.

    1991-01-01

    Deformation within the North American plate in response to various tectonic processes is modeled using an elastic finite element analysis. The tectonic processes considered in the modeling include ridge forces associated with the normal thermal evolution of oceanic lithosphere, shear and normal stresses transmitted across transforms, normal stresses transmitted across convergent boundaries, stresses due to horizontal density contrasts within the continent, and shear tractions applied along the base of the plate. Model stresses are calculated with respect to a lithostatic reference stress state. Shear stresses transmitted across transform boundaries along the San Andreas and Caribbean are small, of the order of 5-10 MPa. Also, compressive stresses of the order of 5-10 MPa transmitted across the major transforms improve the fit to the data. Compressive stresses across convergent margins along the Aleutians and the Middle America trench are important.

  16. Bipolar battery plate

    NASA Technical Reports Server (NTRS)

    Rowlette, John J. (Inventor)

    1985-01-01

    A liquid-impermeable plate (10) having throughplate conductivity with essentially zero resistance comprises an insulator sheet (12) having a series of spaced perforations (14) each of which contains a metal element (16) sealingly received into the perforation (14). A low-cost plate can readily be manufactured by punching a thermoplastic sheet (40) such as polypropylene with a punching tool (52), filling the apertures with lead spheres (63) having a diameter smaller than the holes (50) but larger than the thickness of the sheet, sweeping excess spheres (62) off the sheet with a doctor blade (60) and then pressing a heated platen (74) onto the sheet to swage the spheres into a cylindrical shape and melt the surrounding resin to form a liquid-impermeable collar (4) sealing the metal into the sheet.

  17. Microchannel plate streak camera

    DOEpatents

    Wang, C.L.

    1984-09-28

    An improved streak camera in which a microchannel plate electron multiplier is used in place of or in combination with the photocathode used in prior streak cameras. The improved streak camera is far more sensitive to photons (uv to gamma-rays) than the conventional x-ray streak camera which uses a photocathode. The improved streak camera offers gamma-ray detection with high temporal resolution. It also offers low-energy x-ray detection without attenuation inside the cathode. Using the microchannel plate in the improved camera has resulted in a time resolution of about 150 ps, and has provided a sensitivity sufficient for 1000 keV x-rays.

  18. Microchannel plate streak camera

    DOEpatents

    Wang, C.L.

    1989-03-21

    An improved streak camera in which a microchannel plate electron multiplier is used in place of or in combination with the photocathode used in prior streak cameras is disclosed. The improved streak camera is far more sensitive to photons (UV to gamma-rays) than the conventional x-ray streak camera which uses a photocathode. The improved streak camera offers gamma-ray detection with high temporal resolution. It also offers low-energy x-ray detection without attenuation inside the cathode. Using the microchannel plate in the improved camera has resulted in a time resolution of about 150 ps, and has provided a sensitivity sufficient for 1,000 KeV x-rays. 3 figs.

  19. Uccle Carte du Ciel Plate Catalogue Present in the WFPDB

    NASA Astrophysics Data System (ADS)

    Tsvetkova, Katya; Tsvetkov, Milcho; Lampens, Patricia; Duval, David

    2007-08-01

    We present the catalogue of the Carte du Ciel plates collected at the Royal Observatory of Belgium (ROB) and incorporated in the Wide-Field Plate Database. The catalogue comprises the descriptive information for 682 plates obtained with the Gautier 0.33-m equatorial telescope in the framework of the Carte du Ciel project in the period 1908-1939. The plates were taken using triple exposures with duration of 15 to 30 minutes. An analysis of the ROB CdC catalogue's content is presented. The catalogue, as well as the plate previews taken with a flatbed scanner AGFA (model DUOSCAN HiD) with a resolution of 250 dpi in TIFF format (of size 2.5 MB), are available on-line at http://www.skyarchive.org/.

  20. On the strain energy of laminated composite plates

    NASA Technical Reports Server (NTRS)

    Atilgan, Ali R.; Hodges, Dewey H.

    1991-01-01

    The present effort to obtain the asymptotically correct form of the strain energy in inhomogeneous laminated composite plates proceeds from the geometrically nonlinear elastic theory-based three-dimensional strain energy by decomposing the nonlinear three-dimensional problem into a linear, through-the-thickness analysis and a nonlinear, two-dimensional analysis analyzing plate formation. Attention is given to the case in which each lamina exhibits material symmetry about its middle surface, deriving closed-form analytical expressions for the plate elastic constants and the displacement and strain distributions through the plate's thickness. Despite the simplicity of the plate strain energy's form, there are no restrictions on the magnitudes of displacement and rotation measures.

  1. Mechanisms in Thermal Mechanical Forming of Plates.

    DTIC Science & Technology

    1987-05-01

    specific locations within the plate is discussed, and recommendations for further research are made. Keywords: Metal plates; Ship plates; Material forming; Thermomechanics; Edge effect ; Laser line heating.

  2. The Plate Overlap Technique.

    DTIC Science & Technology

    1978-07-31

    INTRODUCTION 1 II. NOTATION 2 III. THE GNOMONIC PROJECTION 4 IV . THE PLATE OVERLAP TECHNIQUE 6 A. MOTIVATION 6 B. FORNULATION 9 C. ON STATISTICAL RIGOR 14 D...and new hardware. Since this aim was clearly recognized long ago, wherever possible in earlier documents or software development flexibility was...reader should see 1, 2, and 3. The procedures one should use to update stellar positions are discussed in 4 with applica- tions to the SAOC in 5. Non

  3. Plated wire memory subsystem

    NASA Technical Reports Server (NTRS)

    Reynolds, L.; Tweed, H.

    1972-01-01

    The work performed entailed the design, development, construction and testing of a 4000 word by 18 bit random access, NDRO plated wire memory for use in conjunction with a spacecraft imput/output unit and central processing unit. The primary design parameters, in order of importance, were high reliability, low power, volume and weight. A single memory unit, referred to as a qualification model, was delivered.

  4. Elastic plate spallation

    NASA Technical Reports Server (NTRS)

    Oline, L.; Medaglia, J.

    1972-01-01

    The dynamic finite element method was used to investigate elastic stress waves in a plate. Strain displacement and stress strain relations are discussed along with the stiffness and mass matrix. The results of studying point load, and distributed load over small, intermediate, and large radii are reported. The derivation of finite element matrices, and the derivation of lumped and consistent matrices for one dimensional problems with Laplace transfer solutions are included. The computer program JMMSPALL is also included.

  5. Smov FOS Plate Scale

    NASA Astrophysics Data System (ADS)

    Kinney, Anne

    1994-01-01

    The goal is to measure the precise plate scale and orientation. This will be acheived by performing a raster step and dwell sequence in the 4.3 arcsec aperture. The edges of the aperture should be avoided to prevent vignetting effects. An aperture map is required at each step of the dwell sequence. This test has to be conducted for both the RED and BLUE detectors. We will also determine the offset between the two detectors.

  6. Electronic Equipment Cold Plates

    DTIC Science & Technology

    1976-04-01

    equations for such a flow regiae. For laainar flow and Moderate teaperature differwwe« between the well «nd coolant, a aodifled Sieder -Tate...con- figuration. The heat-transfer coefficients, therefore, were determined by using both the Sieder -Tate and McAdams equations and the coaputed...values used In the analytical predictions. As with th* previous cold Plates, the Sieder -Tate equation gave too low of values for the heat- transfer

  7. Martian plate tectonics

    NASA Astrophysics Data System (ADS)

    Sleep, N. H.

    1994-03-01

    The northern lowlands of Mars have been produced by plate tectonics. Preexisting old thick highland crust was subducted, while seafloor spreading produced thin lowland crust during late Noachian and Early Hesperian time. In the preferred reconstruction, a breakup margin extended north of Cimmeria Terra between Daedalia Planum and Isidis Planitia where the highland-lowland transition is relatively simple. South dipping subduction occured beneath Arabia Terra and east dipping subduction beneath Tharsis Montes and Tempe Terra. Lineations associated with Gordii Dorsum are attributed to ridge-parallel structures, while Phelegra Montes and Scandia Colles are interpreted as transfer-parallel structures or ridge-fault-fault triple junction tracks. Other than for these few features, there is little topographic roughness in the lowlands. Seafloor spreading, if it occurred, must have been relatively rapid. Quantitative estimates of spreading rate are obtained by considering the physics of seafloor spreading in the lower (approx. 0.4 g) gravity of Mars, the absence of vertical scarps from age differences across fracture zones, and the smooth axial topography. Crustal thickness at a given potential temperature in the mantle source region scales inversely with gravity. Thus, the velocity of the rough-smooth transition for axial topography also scales inversely with gravity. Plate reorganizations where young crust becomes difficult to subduct are another constraint on spreading age. Plate tectonics, if it occurred, dominated the thermal and stress history of the planet. A geochemical implication is that the lower gravity of Mars allows deeper hydrothermal circulation through cracks and hence more hydration of oceanic crust so that more water is easily subducted than on the Earth. Age and structural relationships from photogeology as well as median wavelength gravity anomalies across the now dead breakup and subduction margins are the data most likely to test and modify hypotheses

  8. Plate motion and deformation

    SciTech Connect

    Minster, B.; Prescott, W.; Royden, L.

    1991-02-01

    Our goal is to understand the motions of the plates, the deformation along their boundaries and within their interiors, and the processes that control these tectonic phenomena. In the broadest terms, we must strive to understand the relationships of regional and local deformation to flow in the upper mantle and the rheological, thermal and density structure of the lithosphere. The essential data sets which we require to reach our goal consist of maps of current strain rates at the earth's surface and the distribution of integrated deformation through time as recorded in the geologic record. Our success will depend on the effective synthesis of crustal kinematics with a variety of other geological and geophysical data, within a quantitative theoretical framework describing processes in the earth's interior. Only in this way can we relate the snapshot of current motions and earth structure provided by geodetic and geophysical data with long-term processes operating on the time scales relevant to most geological processes. The wide-spread use of space-based techniques, coupled with traditional geological and geophysical data, promises a revolution in our understanding of the kinematics and dynamics of plate motions over a broad range of spatial and temporal scales and in a variety of geologic settings. The space-based techniques that best address problems in plate motion and deformation are precise space-geodetic positioning -- on land and on the seafloor -- and satellite acquisition of detailed altimetric and remote sensing data in oceanic and continental areas. The overall science objectives for the NASA Solid Earth Science plan for the 1990's, are to Understand the motion and deformation of the lithosphere within and across plate boundaries'', and to understand the dynamics of the mantle, the structure and evolution of the lithosphere, and the landforms that result from local and regional deformation. 57 refs., 7 figs., 2 tabs.

  9. Armor Plate Surface Roughness Measurements

    DTIC Science & Technology

    2005-04-01

    Armor Plate Surface Roughness Measurements by Brian Stanton, William Coburn, and Thomas J. Pizzillo ARL-TR-3498 April 2005... Armor Plate Surface Roughness Measurements Brian Stanton, William Coburn and Thomas J. Pizzillo Sensors and Electron Devices Directorate...October 2004 5a. CONTRACT NUMBER 5b. GRANT NUMBER 4. TITLE AND SUBTITLE Armor Plate Surface Roughness Measurements 5c. PROGRAM ELEMENT NUMBER

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

    PubMed

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

    2016-03-01

    Microtiter plates (MTP) and automatized techniques are increasingly applied in the field of biotechnology. However, the susceptibility of MTPs to edge effects such as thermal gradients can lead to high variation of measured enzyme activities. In an effort to enhance experimental reliability, to quantify, and to minimize instrument-caused deviations in enzyme kinetics between two MTP-readers, we comprehensively quantified temperature distribution in 96-well MTPs. We demonstrated the robust application of the absorbance dye cresol red as easily applicable temperature indicator in cuvettes and MTPs and determined its accuracy to ±0.16°C. We then quantified temperature distributions in 96-well MTPs revealing temperature deviations over single MTP of up to 2.2°C and different patterns in two commercial devices (BioTek Synergy 4 and Synergy Mx). The obtained liquid temperature was shown to be substantially controlled by evaporation. The temperature-induced enzyme activity variation within MTPs amounted to about 20 %. Activity deviations between MTPs and to those in cuvettes were determined to 40 % due to deviations from the set temperature in MTPs. In conclusion, we propose a better control of experimental conditions in MTPs or alternative experimental systems for reliable determination of kinetic parameters for bioprocess development. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Buckling of trapezoidal plates

    SciTech Connect

    Radloff, H.D.; Hyer, M.W.; Nemeth, M.P.

    1995-12-31

    This study focuses on the buckling response of flat composite plates with an isosceles planform, as shown below. The study consisted of both analytical and experimental phases. In the study the parallel edges x = 0 and L were clamped or simply supported. Clamped parallel edges were used in the experimental phase of the study. The nonparallel edges were simply supported. The parallel edges were assumed to remain straight and buckling was assumed to occur because these edges moved toward each other. A Rayleigh-Ritz approach was used, with the finite-element code ABAQUS being used to check specific cases. The buckling displacements were assumed to be in the form of harmonic functions. The prebuckling force resultant N{sub x} was assumed to be given by N{sub x}(x,y) = P/W(x), where P is the applied load and W(x) is the varying width of the plate. N{sub y}(x,y) and N{sub x,y}(x,y) were taken to be zero. Experiments were conducted on a number of laminates and a number of plate geometries, and the results were compared to predictions of the Rayleigh-Ritz scheme. In general correlation was good, though the analysis underpredicted the number of buckling half-waves.

  12. Origin of Small Tectonic Plates

    NASA Astrophysics Data System (ADS)

    Mallard, C.; Coltice, N.; Seton, M.; Müller, D.; Tackley, P.

    2015-12-01

    The plate tectonic theory allowed to split the Earth surface into 6 (Le Pichon 1968) to 52 tectonic plates (Bird 2003). These plates are separated into two groups: the first of 7 large plates and the second of numerous smaller plates (Morra et al 2013). Previous studies using the reconstruction of the past 200 My, suggest that the size of large plates is driven by mantle flow. But the tools employed are descriptive (Morra et al 2013, Sornette and Pisarenko 2003), hence ignoring forces and physical principles within the lithosphere and the mantle. The processes at the origin of small plates remain unknown. We developed a new approach to explain the plate sizes. We demonstrate that the physics of convection drives it. We applied plate tectonics theory on 3D spherical convection models generating plate-like motions, which give access to a complete survey of data: velocities, viscosity and heat flow. Our data show that (1) the large plates depend on the dominating scale of the convective flow due to the initiation or the shutdown of subductions; (2) the smaller plates are generated thanks to large variability of regional stresses along subduction zone by slab pull and suction influenced by the geometry of trenches. Our results are consistent with the quick reorganizations of back-arc basins occuring synchronously with the modification of subduction zones geometry around the Pacific plate (Sdrolias et al 2004). Hence, we conclude that (1) the decreasing number of small plates in the plate reconstructions back in time is an artifact induced by their short lifetime, that is why they are artificially ignored; (2) the geometry of past trenches is simplified leading to an underestimation of the length of subduction zones.

  13. Characteristics and applications of advanced technology microchannel plates

    NASA Astrophysics Data System (ADS)

    Horton, J. R.; Tasker, G. W.; Fijol, J. J.

    1990-09-01

    A method for fabrication of novel thin-film continuous dynode electron multipliers is described. The feasibility of crucial manufacturing steps, including anisotropic dry etching of substrates into photolithographically-defined arrays of high-aspect-ratio channels, and the formation of thin-film continuous dynodes by CVD is shown. Potential performance and design advantages of this advanced technology-microchannel plate over the conventional reduced lead silicate glass microchannel plate and implications for new applications are discussed.

  14. Iberian plate kinematics: A jumping plate boundary between Eurasia and Africa

    USGS Publications Warehouse

    Srivastava, S.P.; Schouten, Hans; Roest, W.R.; Klitgord, Kim D.; Kovacs, L.C.; Verhoef, J.; Macnab, R.

    1990-01-01

    THE rotation of Iberia and its relation to the formation of the Pyrenees has been difficult to decipher because of the lack of detailed sea-floor spreading data, although several models have been proposed1-7. Here we use detailed aeromagnetic measurements from the sea floor offshore of the Grand Banks of Newfoundland to show that Iberia moved as part of the African plate from late Cretaceous to mid-Eocene time, with a plate boundary extending westward from the Bay of Biscay. When motion along this boundary ceased, a boundary linking extension in the King's Trough to compression along the Pyrenees came into existence. Finally, since the late Oligocene, Iberia has been part of the Eurasian plate, with the boundary between Eurasia and Africa situated along the Azores-Gibraltar fracture zone.

  15. Symmetries in laminated composite plates

    NASA Technical Reports Server (NTRS)

    Noor, A. K.

    1976-01-01

    The different types of symmetry exhibited by laminated anisotropic fibrous composite plates are identified and contrasted with the symmetries of isotropic and homogeneous orthotropic plates. The effects of variations in the fiber orientation and the stacking sequence of the layers on the symmetries exhibited by composite plates are discussed. Both the linear and geometrically nonlinear responses of the plates are considered. A simple procedure is presented for exploiting the symmetries in the finite element analysis. Examples are given of square, skew and polygonal plates where use of symmetry concepts can significantly reduce the scope and cost of analysis.

  16. Plate tectonics conserves angular momentum

    NASA Astrophysics Data System (ADS)

    Bowin, C.

    2009-03-01

    A new combined understanding of plate tectonics, Earth internal structure, and the role of impulse in deformation of the Earth's crust is presented. Plate accelerations and decelerations have been revealed by iterative filtering of the quaternion history for the Euler poles that define absolute plate motion history for the past 68 million years, and provide an unprecedented precision for plate angular rotation variations with time at 2-million year intervals. Stage poles represent the angular rotation of a plate's motion between adjacent Euler poles, and from which the maximum velocity vector for a plate can be determined. The consistent maximum velocity variations, in turn, yield consistent estimates of plate accelerations and decelerations. The fact that the Pacific plate was shown to accelerate and decelerate, implied that conservation of plate tectonic angular momentum must be globally conserved, and that is confirmed by the results shown here (total angular momentum ~1.4 E+27 kgm2s-1). Accordingly, if a plate decelerates, other plates must increase their angular momentums to compensate. In addition, the azimuth of the maximum velocity vectors yields clues as to why the "bend" in the Emperor-Hawaiian seamount trend occurred near 46 Myr. This report summarizes processing results for 12 of the 14 major tectonic plates of the Earth (except for the Juan de Fuca and Philippine plates). Plate accelerations support the contention that plate tectonics is a product of torques that most likely are sustained by the sinking of positive density anomalies due to phase changes in subducted gabbroic lithosphere at depth in the upper lower mantle (above 1200 km depth). The tectonic plates are pulled along by the sinking of these positive mass anomalies, rather than moving at near constant velocity on the crests of convection cells driven by rising heat. These results imply that spreading centers are primarily passive reactive features, and fracture zones (and wedge-shaped sites

  17. Plate fixation of prostheses after segmental resection for bone tumours.

    PubMed

    Coathup, M J; Cobb, J P; Walker, P S; Blunn, G W

    2000-11-01

    This study investigated the concept of using plates to attach endoprostheses to bone after segmental resection for bone tumours in an animal model. Titanium alloy plates integrated with the prosthesis and coated with hydroxyapatite were attached to bone by screws. This type of uncemented fixation relied on the induction of periosteal bone formation into and around the plates to secure the implant to bone. Two, three, and six-slotted plate designs were investigated. On retrieval, each plate was securely fixed by new bone. Bone apposition on the external surface of the plates occurred through a combination of periosteal bone production, invasion of bone through slots in the plate, and bone growth over the ends of the plates. Most plates became incorporated into a remodelled cortex. Higher bone turnover rates (microm day(-1)) were seen in bone in the slots of the plate compared with normal cortical bone turnover (p < 0.05). Significantly higher rates of turnover were measured beneath slotted parts of the plates compared with regions below the unslotted parts (p < 0.05). The cross-sectional area of bone surrounding the six-plate implant design was significantly higher than that of the three-plate (p < 0.05) and two-plate (p < 0.05) designs. In addition, significantly more bone formed adjacent to the six-plated implant design compared with that in the contralateral limb (p = 0.002). However, no significant difference was found when the total cortical area around the three-plated design was compared with that of the contralateral limb (p = 0.63). In contrast, significantly less bone was measured adjacent to the two-plate design than in the untreated limb (p = 0.001). Image analysis also demonstrated increased cortical porosity adjacent to the six-plate design compared with the three-plate (p = 0.004) and two-plate (p < 0.05) designs. Finite element analysis demonstrated that the six and three-plate designs increased the second moment of area compared with that in the

  18. Reducing Plate Tectonic Misconceptions with Lecture Tutorials

    NASA Astrophysics Data System (ADS)

    Kortz, K. M.; Smay, J. M.; Mattera, A. V.; Clark, S. K.

    2009-12-01

    In order to address student difficulties with and common misconceptions about plate tectonics, we created five Lecture Tutorials suitable for introductory geoscience courses. Lecture Tutorials are 10-15 minute worksheets that students complete in class in small groups to make learning more student-centered. Students build their knowledge with questions that progressively become more difficult, requiring them think about their misconceptions. Our research indicates that the Lecture Tutorials successfully decrease student misconceptions. For example, few introductory students identify the mantle wedge as the location of melting at subduction zones. Instead, students frequently think melting occurs at the trench, in magma chambers within volcanoes, or where images commonly show the subducting slab disappearing. One of the Lecture Tutorials helps the students determine why melting occurs and therefore identify the correct locations of melting at convergent boundaries, divergent boundaries, and hotspots. This Lecture Tutorial includes a hypothetical “debate” with statements expressing the misconceptions and one expressing the correct scientific idea of where melting occurs. Students are asked to explain why they agree with one of the statements, so they must directly think about any misconceptions they may have. Additional difficulties addressed by the Lecture Tutorials include identification of the direction of plate movement at ocean ridges and the locations and formation of basic plate tectonic features, such as trenches, volcanoes, ocean ridges, and plate boundaries. After instruction, students completed questionnaires that probed their understanding of plate tectonics, and students who completed the Lecture Tutorials performed significantly better on relevant questions. For example, when asked to circle the locations on a diagram where melting occurred, students who completed the Lecture Tutorials correctly circled the mantle wedge more often than other

  19. Fuel cell end plate structure

    DOEpatents

    Guthrie, Robin J.; Katz, Murray; Schroll, Craig R.

    1991-04-23

    The end plates (16) of a fuel cell stack (12) are formed of a thin membrane. Pressure plates (20) exert compressive load through insulation layers (22, 26) to the membrane. Electrical contact between the end plates (16) and electrodes (50, 58) is maintained without deleterious making and breaking of electrical contacts during thermal transients. The thin end plate (16) under compressive load will not distort with a temperature difference across its thickness. Pressure plate (20) experiences a low thermal transient because it is insulated from the cell. The impact on the end plate of any slight deflection created in the pressure plate by temperature difference is minimized by the resilient pressure pad, in the form of insulation, therebetween.

  20. Hypervelocity impact on shielded plates

    NASA Technical Reports Server (NTRS)

    Smith, James P.

    1993-01-01

    A ballistic limit equation for hypervelocity impact on thin plates is derived analytically. This equation applies to cases of impulsive impact on a plate that is protected by a multi-shock shield, and it is valid in the range of velocity above 6 km/s. Experimental tests were conducted at the NASA Johnson Space Center on square aluminum plates. Comparing the center deflections of these plates with the theoretical deflections of a rigid-plastic plate subjected to a blast load, one determines the dynamic yield strength of the plate material. The analysis is based on a theory for the expansion of the fragmented projectile and on a simple failure criterion. Curves are presented for the critical projectile radius versus the projectile velocity, and for the critical plate thickness versus the velocity. These curves are in good agreement with curves that have been generated empirically.

  1. Strain Library Imaging Protocol for high-throughput, automated single-cell microscopy of large bacterial collections arrayed on multiwell plates.

    PubMed

    Shi, Handuo; Colavin, Alexandre; Lee, Timothy K; Huang, Kerwyn Casey

    2017-02-01

    Single-cell microscopy is a powerful tool for studying gene functions using strain libraries, but it suffers from throughput limitations. Here we describe the Strain Library Imaging Protocol (SLIP), which is a high-throughput, automated microscopy workflow for large strain collections that requires minimal user involvement. SLIP involves transferring arrayed bacterial cultures from multiwell plates onto large agar pads using inexpensive replicator pins and automatically imaging the resulting single cells. The acquired images are subsequently reviewed and analyzed by custom MATLAB scripts that segment single-cell contours and extract quantitative metrics. SLIP yields rich data sets on cell morphology and gene expression that illustrate the function of certain genes and the connections among strains in a library. For a library arrayed on 96-well plates, image acquisition can be completed within 4 min per plate.

  2. Gas chromatography fractionation platform featuring parallel flame-ionization detection and continuous high-resolution analyte collection in 384-well plates.

    PubMed

    Jonker, Willem; Clarijs, Bas; de Witte, Susannah L; van Velzen, Martin; de Koning, Sjaak; Schaap, Jaap; Somsen, Govert W; Kool, Jeroen

    2016-09-02

    Gas chromatography (GC) is a superior separation technique for many compounds. However, fractionation of a GC eluate for analyte isolation and/or post-column off-line analysis is not straightforward, and existing platforms are limited in the number of fractions that can be collected. Moreover, aerosol formation may cause serious analyte losses. Previously, our group has developed a platform that resolved these limitations of GC fractionation by post-column infusion of a trap solvent prior to continuous small-volume fraction collection in a 96-wells plate (Pieke et al., 2013 [17]). Still, this GC fractionation set-up lacked a chemical detector for the on-line recording of chromatograms, and the introduction of trap solvent resulted in extensive peak broadening for late-eluting compounds. This paper reports advancements to the fractionation platform allowing flame ionization detection (FID) parallel to high-resolution collection of a full GC chromatograms in up to 384 nanofractions of 7s each. To this end, a post-column split was incorporated which directs part of the eluate towards FID. Furthermore, a solvent heating device was developed for stable delivery of preheated/vaporized trap solvent, which significantly reduced band broadening by post-column infusion. In order to achieve optimal analyte trapping, several solvents were tested at different flow rates. The repeatability of the optimized GC fraction collection process was assessed demonstrating the possibility of up-concentration of isolated analytes by repetitive analyses of the same sample. The feasibility of the improved GC fractionation platform for bioactivity screening of toxic compounds was studied by the analysis of a mixture of test pesticides, which after fractionation were subjected to a post-column acetylcholinesterase (AChE) assay. Fractions showing AChE inhibition could be unambiguously correlated with peaks from the parallel-recorded FID chromatogram. Copyright © 2016 Elsevier B.V. All rights

  3. Plated wire memory subsystem

    NASA Technical Reports Server (NTRS)

    Carpenter, K. H.

    1974-01-01

    The design, construction, and test history of a 4096 word by 18 bit random access NDRO Plated Wire Memory for use in conjunction with a spacecraft input/output and central processing unit is reported. A technical and functional description is given along with diagrams illustrating layout and systems operation. Test data is shown on the procedures and results of system level and memory stack testing, and hybrid circuit screening. A comparison of the most significant physical and performance characteristics of the memory unit versus the specified requirements is also included.

  4. Shuttle plate braiding machine

    NASA Technical Reports Server (NTRS)

    Huey, Jr., Cecil O. (Inventor)

    1994-01-01

    A method and apparatus for moving yarn in a selected pattern to form a braided article. The apparatus includes a segmented grid of stationary support elements and a plurality of shuttles configured to carry yarn. The shuttles are supported for movement on the grid assembly and each shuttle includes a retractable plunger for engaging a reciprocating shuttle plate that moves below the grid assembly. Such engagement at selected times causes the shuttles to move about the grid assembly in a selected pattern to form a braided article of a particular geometry.

  5. Quantitative analysis of changes in actin microfilament contribution to cell plate development in plant cytokinesis

    PubMed Central

    Higaki, Takumi; Kutsuna, Natsumaro; Sano, Toshio; Hasezawa, Seiichiro

    2008-01-01

    Background Plant cells divide by the formation of new cross walls, known as cell plates, from the center to periphery of each dividing cell. Formation of the cell plate occurs in the phragmoplast, a complex structure composed of membranes, microtubules (MTs) and actin microfilaments (MFs). Disruption of phragmoplast MTs was previously found to completely inhibit cell plate formation and expansion, indicative of their crucial role in the transport of cell plate membranes and materials. In contrast, disruption of MFs only delays cell plate expansion but does not completely inhibit cell plate formation. Despite such findings, the significance and molecular mechanisms of MTs and MFs remain largely unknown. Results Time-sequential changes in MF-distribution were monitored by live imaging of tobacco BY-2 cells stably expressing the GFP-actin binding domain 2 (GFP-ABD2) fusion protein, which vitally co-stained with the endocytic tracer, FM4-64, that labels the cell plate. During cytokinesis, MFs accumulated near the newly-separated daughter nuclei towards the emerging cell plate, and subsequently approached the expanding cell plate edges. Treatment with an actin polymerization inhibitor caused a decrease in the cell plate expansion rate, which was quantified using time-lapse imaging and regression analysis. Our results demonstrated time-sequential changes in the contribution of MFs to cell plate expansion; MF-disruption caused about a 10% decrease in the cell plate expansion rate at the early phase of cytokinesis, but about 25% at the late phase. MF-disruption also caused malformation of the emerging cell plate at the early phase, indicative of MF involvement in early cell plate formation and expansion. The dynamic movement of endosomes around the cell plate was also inhibited by treatment with an actin polymerization inhibitor and a myosin ATPase inhibitor, respectively. Furthermore, time-lapse imaging of the endoplasmic reticulum (ER) revealed that MFs were involved in

  6. The influence of subminimal inhibitory concentrations of benzalkonium chloride on biofilm formation by Listeria monocytogenes.

    PubMed

    Ortiz, Sagrario; López, Victoria; Martínez-Suárez, Joaquín V

    2014-10-17

    Disinfectants, such as benzalkonium chloride (BAC), are commonly used to control Listeria monocytogenes and other pathogens in food processing plants. Prior studies have demonstrated that the resistance to BAC of L. monocytogenes was associated with the prolonged survival of three strains of molecular serotype 1/2a in an Iberian pork processing plant. Because survival in such environments is related to biofilm formation, we hypothesised that the influence of BAC on the biofilm formation potential of L. monocytogenes might differ between BAC-resistant strains (BAC-R, MIC≥10mg/L) and BAC-sensitive strains (BAC-S, MIC≤2.5mg/L). To evaluate this possibility, three BAC-R strains and eight BAC-S strains, which represented all of the molecular serotype 1/2a strains detected in the sampled plant, were compared. Biofilm production was measured using the crystal violet staining method in 96-well microtitre plates. The BAC-R strains produced significantly (p<0.05) less biofilm than the BAC-S in the absence of BAC, independent of the rate of planktonic growth. In contrast, when the biofilm values were measured in the presence of BAC, one BAC-R strain (S10-1) was able to form biofilm at 5mg/L of BAC, which prevented biofilm formation among the rest of the strains. A genetic determinant of BAC resistance recently described in L. monocytogenes (Tn6188) was detected in S10-1. When a BAC-S strain and its spontaneous mutant BAC-R derivative were compared, resistance to BAC led to biofilm formation at 5mg/L of BAC and to a significant (p<0.05) stimulation of biofilm formation at 1.25mg/L of BAC, which significantly (p<0.05) reduced the biofilm level in the parent BAC-S strain. Our results suggest that the effect of subminimal inhibitory concentrations of BAC on biofilm production by L. monocytogenes might differ between strains with different MICs and even between resistant strains with similar MICs but different genetic determinants of BAC resistance. For BAC-R strains similar

  7. Earth's Decelerating Tectonic Plates (Invited)

    NASA Astrophysics Data System (ADS)

    Forte, A. M.; Moucha, R.; Rowley, D. B.; Quere, S.; Mitrovica, J. X.; Simmons, N. A.; Grand, S. P.

    2009-12-01

    We employ a recently developed convection model constrained by seismic, geodynamic and mineral physics data (Simmons et al., GJI 2009) to explore the impact of time-dependent changes in mantle buoyancy forces on tectonic plate accelerations. This plate-coupled mantle convection model incorporates a viscosity structure that reconciles both glacial isostatic adjustment and global convection-related data sets (Mitrovica & Forte, EPSL 2004) and it successfully reproduces present-day plate velocities, global surface gravity and topography data. This convection model predicts the recent deceleration of several major plates in the Pacific and Indo-Atlantic hemispheres. Independent verification of these predictions is a fundamental test of the plausibility of the buoyancy forces and rheological structure in the convection model. To this end, we consider marine magnetic anomaly and space geodetic constraints on tectonic plate motions to determine a new global map of present-day rates of change of plate velocities. This map shows that several major plates, such as the Pacific, Africa and Nazca plates are presently decelerating and that they contribute to a globally-averaged slowdown in tectonic plate speeds. These joint geologic-geodetic inferences of plate decelerations are consistent with those predicted by our tomography-based convection model.

  8. Survey of ion plating sources. [conferences

    NASA Technical Reports Server (NTRS)

    Spalvins, T.

    1979-01-01

    Based on the type of evaporation source, gaseous media and mode of transport, the following is discussed: resistance, electron beam, sputtering, reactive and ion beam evaporation. Ionization efficiencies and ion energies in the glow discharge determine the percentage of atoms which are ionized under typical ion plating conditions. The plating flux consists of a small number of energetic ions and a large number of energetic neutrals. The energy distribution ranges from thermal energies up to a maximum energy of the discharge. The various reaction mechanisms which contribute to the exceptionally strong adherence - formation of a graded sustrate/coating interface are not fully understood, however the controlling factors are evaluated. The influence of process variables on the nucleation and growth characteristics are illustrated in terms of morphological changes which affect the mechanical and tribological properties of the coating.

  9. Ionospheric plasma flow over large high-voltage space platforms. I - Ion-plasma-time scale interactions of a plate at zero angle of attack. II - The formation and structure of plasma wake

    NASA Technical Reports Server (NTRS)

    Wang, J.; Hastings, D. E.

    1992-01-01

    The paper presents the theory and particle simulation results for the ionospheric plasma flow over a large high-voltage space platform at a zero angle of attack and at a large angle of attack. Emphasis is placed on the structures in the large, high-voltage regime and the transient plasma response on the ion-plasma time scale. Special consideration is given to the transient formation of the space-charge wake and its steady-state structure.

  10. Multiscale modeling of growth plate cartilage mechanobiology.

    PubMed

    Gao, Jie; Williams, John L; Roan, Esra

    2017-04-01

    Growth plate chondrocytes are responsible for bone growth through proliferation and differentiation. However, the way they experience physiological loads and regulate bone formation, especially during the later developmental phase in the mature growth plate, is still under active investigation. In this study, a previously developed multiscale finite element model of the growth plate is utilized to study the stress and strain distributions within the cartilage at the cellular level when rapidly compressed to 20 %. Detailed structures of the chondron are included in the model to examine the hypothesis that the same combination of mechanoregulatory signals shown to maintain cartilage or stimulate osteogenesis or fibrogenesis in the cartilage anlage or fracture callus also performs the same function at the cell level within the chondrons of growth plate cartilage. Our cell-level results are qualitatively and quantitatively in agreement with tissue-level theories when both hydrostatic cellular stress and strain are considered simultaneously in a mechanoregulatory phase diagram similar to that proposed at the tissue level by Claes and Heigele for fracture healing. Chondrocytes near the reserve/proliferative zone border are subjected to combinations of high compressive hydrostatic stresses ([Formula: see text] MPa), and cell height and width strains of [Formula: see text] to [Formula: see text] respectively, that maintain cartilage and keep chondrocytes from differentiating and provide conditions favorable for cell division, whereas chondrocytes closer to the hypertrophic/calcified zone undergo combinations of lower compressive hydrostatic stress ([Formula: see text] MPa) and cell height and width strains as low as [Formula: see text] to +4 %, respectively, that promote cell differentiation toward osteogenesis; cells near the outer periphery of the growth plate structure experience a combination of low compressive hydrostatic stress (0 to [Formula: see text] MPa) and

  11. Plating on difficult-to-plate metals: what's new

    SciTech Connect

    Wiesner, H.J.

    1980-07-30

    Some of the changes since 1970 in procedures for plating on such materials as titanium, molybdenum, silicon, aluminum, and gallium arsenide are summarized. While basic procedures for plating some of these materials were developed as many as 30 to 40 years ago, changes in the end uses of the plated products have necessitated new plating processes. In some cases, vacuum techniques - such as ion bombardment, ion implantation, and vacuum metallization - have been introduced to improve the adhesion of electrodeposits. In other cases, these techniques have been used to deposit materials upon which electrodeposits are required.

  12. Plate failure following plate osteosynthesis in periprosthetic femoral fractures.

    PubMed

    Boesmueller, Sandra; Baumbach, Sebastian F; Hofbauer, Marcus; Wozasek, Gerald E

    2015-10-01

    Increasing numbers of total knee and hip arthroplasties result in a growing number of periprosthetic femoral fractures (PPFF). PPFF with a stable stem component are treated commonly with plate osteosynthesis. Therefore plate failure is seen as a major complication. The aim of this retrospective study was to investigate the patients' outcome after plate failure. The database of a Level 1 trauma center was searched for all patients treated for a PPFF with plate osteosynthesis. Patients with plate failure were investigated specifically. Standard demographic data, details on initial arthroplasty, trauma, and treatment were recorded for all patients. All fractures were classified and their outcome reviewed. Seven (8.8%) out of 80 patients treated with plate osteosynthesis following PPFF met our inclusion criterion being plate failure. All these patients were female, with an average age at primary surgery of 74 ± 13 years and a mean follow-up of 885 days (range, 264-2549). Four patients suffered a PPFF after total hip arthroplasty (THA) (2 Vancouver Type B1 and 2 Type C) and three after total knee arthroplasty (TKA) (Lewis-Rorabeck Type II). Following plate failure, four patients healed uneventfully and three patients experienced complications such as pseudarthrosis, screw loosening, and further plate failure. In patients with poor bone quality, bone graft, bone cement, and bone biologics have to be considered in revision surgery. Furthermore, long-stem revision and tumor prosthesis are an additional solution.

  13. Localised Plate Motion on Venus

    NASA Astrophysics Data System (ADS)

    Ghail, R. C.

    1996-03-01

    The volcanic and tectonic features observed in Dali Vinculum, Parga Vinculum and Imdr Regio are concentrated at long, narrow, curvilinear zones, with relatively minor volcanism and tectonism between these zones. These zones, whilst more diffuse than terrestrial plate boundaries, nevertheless define the margins of tectonic plates. In contrast to Earth, however, it appears that venusian plates are neither created nor destroyed by lateral motion. Rather, plates are thinned and intruded at vincula plate boundaries, vertically accreted by small-scale intra-plate (planitia) volcanism and perhaps destroyed by delamination of thickened crust in tesserae and montane regions such as Thetis Regio and Ishtar Terra. The diversity in age both between and within these three areas together with the evidence for infrequent, small scale resurfacing in the planitiae are difficult to reconcile with a non-uniformitarian geological process.

  14. Growth Plate Injuries

    MedlinePlus

    ... where many inherited disorders that affect the musculoskeletal system appear. Scientists are beginning to understand the genes and gene mutations involved in skeletal formation, growth, and development. This new information is ...

  15. Dissolution of HFIR control plates

    SciTech Connect

    Posey, J.C.

    1984-03-01

    A process was developed for the dissolution of High Flux Isotope Reactor (HFIR) control plates. These plates consist of aluminum metal, intensely radioactive europium oxide, and a small amount of tantalum metal. The radioactive solution will be diluted, mixed with grout, and disposed of by shale fracture. The plates are dissolved in nitric acid using a mercury catalyst. Conditions were determined that would produce a reaction rate compatible with existing equipment. 3 references, 1 figure, 3 tables.

  16. Channel plate for DNA sequencing

    DOEpatents

    Douthart, Richard J.; Crowell, Shannon L.

    1998-01-01

    This invention is a channel plate that facilitates data compaction in DNA sequencing. The channel plate has a length, a width and a thickness, and further has a plurality of channels that are parallel. Each channel has a depth partially through the thickness of the channel plate. Additionally an interface edge permits electrical communication across an interface through a buffer to a deposition membrane surface.

  17. Channel plate for DNA sequencing

    DOEpatents

    Douthart, R.J.; Crowell, S.L.

    1998-01-13

    This invention is a channel plate that facilitates data compaction in DNA sequencing. The channel plate has a length, a width and a thickness, and further has a plurality of channels that are parallel. Each channel has a depth partially through the thickness of the channel plate. Additionally an interface edge permits electrical communication across an interface through a buffer to a deposition membrane surface. 15 figs.

  18. Ion plating for the future

    NASA Technical Reports Server (NTRS)

    Spalvins, T.

    1981-01-01

    The ion plating techniques are classified relative to the instrumental set up, evaporation media, and mode of transport. A distinction is drawn between the low vacuum (plasma) and high vacuum (ion beam) techniques. Ion plating technology is discussed at the fundamental and industrial level. At the fundamental level, the capabilities and limitations of the plasma (evaporant flux) and film characteristics are evaluated. And on the industrial level, the performance and potential uses of ion plated films are discussed.

  19. Glass-bead peen plating

    NASA Technical Reports Server (NTRS)

    Graves, J. R.

    1974-01-01

    Peen plating of aluminum, copper, and nickel powders was investigated. Only aluminum was plated successfully within the range of peen plating conditions studied. Optimum plating conditions for aluminum were found to be: (1) bead/powder mixture containing 25 to 35% powder by weight, (2) peening intensity of 0.007A as measured by Almen strip, and (3) glass impact bead diameter of at least 297 microns (0.0117 inches) for depositing-100 mesh aluminum powder. No extensive cleaning or substrate preparation is required beyond removing loose dirt or heavy oil.

  20. True Shear Parallel Plate Viscometer

    NASA Technical Reports Server (NTRS)

    Ethridge, Edwin; Kaukler, William

    2010-01-01

    This viscometer (which can also be used as a rheometer) is designed for use with liquids over a large temperature range. The device consists of horizontally disposed, similarly sized, parallel plates with a precisely known gap. The lower plate is driven laterally with a motor to apply shear to the liquid in the gap. The upper plate is freely suspended from a double-arm pendulum with a sufficiently long radius to reduce height variations during the swing to negligible levels. A sensitive load cell measures the shear force applied by the liquid to the upper plate. Viscosity is measured by taking the ratio of shear stress to shear rate.

  1. Laser-driven flyer plate

    DOEpatents

    Paisley, Dennis L.

    1991-01-01

    Apparatus for producing high velocity flyer plates involving placing a layer of dielectric material between a first metal foil and a second metal foil. With laser irradiation through an optical substrate, the first metal foil forms a plasma in the area of the irradiation, between the substrate and the solid portion of the first metal foil. When the pressure between the substrate and the foil reaches the stress limit of the dielectric, the dielectric will break away and launch the flyer plate out of the second metal foil. The mass of the flyer plate is controlled, as no portion of the flyer plate is transformed into a plasma.

  2. Ballistic Limit of CFRP Plates

    NASA Astrophysics Data System (ADS)

    Higashide, Masumi; Nagao, Yosuke; Kibe, Seishiro; Francesconi, Alessandro; Pavarin, Daniele

    JAXA has carried out the hypervelocity impact tests of carbon fiber reinforced plastic (CFRP) plates together with University of Padova. Quasi-isotropic CFRP plates of 2.3, 3.5, and 4.7 mm in thickness were tested. Aluminum sphere of 0.8 to 2.9 mm in diameter was used as projectiles. With a two-stage light gas gun, the projectile was launched with a velocity range of 2 to 5 km/sec in the normal direction to the CFRP plate. Since the perforated hole and the crater on the CFRP plate after the impact are filled with flakes of the carbon fiber, it is difficult to determine the perforation of the projectile. Therefore, whether the projectile perforated the CFRP plate or not was decided by the craters on a copper plate installed behind the CFRP plate. After the impact, peeling along the fiber direction was observed on the surface of the CFRP plate. Moreover, internal delamination was generated near the surface. Finally, a ballistic limit equation of CFRP plates of 2 to 5 mm in thickness was calculated on the basis of the Cour-Palais equation. The ballistic limit equation was in good agreement with the test results.

  3. Carbon-assisted flyer plates

    SciTech Connect

    Stahl, David B.; Paisley, Dennis L.

    1994-01-01

    A laser driven flyer plate utilizing an optical fiber connected to a laser. The end of the optical fiber has a layer of carbon and a metal layer deposited onto it. The carbon layer provides the laser induced plasma which is superior to the plasma produced from most metals. The carbon layer plasma is capable of providing a flatter flyer plate, converting more of the laser energy to driving plasma, promoting a higher flyer plate acceleration, and providing a more uniform pulse behind the plate. In another embodiment, the laser is in optical communication with a substrate onto which a layer of carbon and a layer of metal have been deposited.

  4. Plate motion controls on back-arc spreading. [Cenozoic movement in Western Pacific

    NASA Technical Reports Server (NTRS)

    Fein, J. B.; Jurdy, D. M.

    1986-01-01

    The motions of the subducting and the overriding plates influence the spatial and temporal distribution of back-arc spreading. Cenozoic plate motions in hot spot-fixed and no-net-rotation reference frames were studied with attention to correlations between changes in motion and episodes of back-arc spreading in the western Pacific. The results suggest that major back-arc opening occurs when both the overriding plate retreats from the trench in an absolute sense and the subducting plate undergoes a significant speed-up. Neither phenomenon alone is sufficient to initiate spreading. Three major plate velocity increases can be identified in the Cenozoic: (1) the Pacific plate 5-9 Ma; (2) the Indian plate at 27 Ma; and (3) the Pacific plate at 43 Ma, due to its shift from northerly to more westerly motion. At the present time, the Indian and Philippine are the only overriding plates that are retreating from their Pacific trenches and back-arc spreading occurs only on these two retreating plates. Although the Indian plate has been retreating for at least 25 Ma, back-arc spreading began only following the Pacific plate speed-up 5-9 Ma. Earlier, during the Indian plate speed-up, no overriding plates were retreating strongly and no back-arc spreading epsiodes are preserved from this time. For the earliest Pacific plate shift at 43 Ma, the Eurasian plate was not advancing, thus creating the only favorable plate kinematic conditions in the Cenozoic for back-arc basin formation in this region. It is unclear whether extension in the Japan Sea is a result of these conditions.

  5. Adaptive Focused Acoustics (AFA) Improves the Performance of Microtiter Plate ELISAs.

    PubMed

    Green, David J; Rudd, Edwin A; Laugharn, James A

    2014-08-01

    We investigated the use of Adaptive Focused Acoustics (AFA) technology to improve the performance of microtiter plate enzyme-linked immunosorbent assays (ELISAs). Experiments were performed with commercially available AFA instrumentation and off-the-shelf 96-well microtiter plate sandwich ELISAs. AFA was applied over a range of acoustic energies, temperatures, and durations to the antigen/antibody binding step of an ELISA for measuring HIV-1 p24 in tissue culture samples. AFA-mediated antigen/antibody binding was enhanced up to 2-fold over passive binding at comparable temperatures and was superior or comparable at low temperature (8-10 °C) to passive binding at 37 °C. Lower nonspecific binding (NSB), lower inter- and intra-assay coefficients of variation (CVs), higher Z' factors, and lower limits of detection (LODs) were measured in AFA-mediated assays compared with conventional passive binding. In a more limited study, AFA enhancement of antigen/antibody binding and lower NSB was measured in an ELISA for measuring IGFBP-3 in human plasma. We conclude from this study that application of AFA to antigen/antibody binding steps in microtiter plate ELISAs can enhance key assay performance parameters, particularly Z' factors and LODs. These features render AFA-mediated binding assays potentially more useful in applications such as high-throughput screening and in vitro diagnostics than assays processed with conventional passive antigen/antibody binding steps.

  6. Scintillator plate calorimetry

    SciTech Connect

    Price, L.E.

    1990-01-01

    Calorimetry using scintillator plates or tiles alternated with sheets of (usually heavy) passive absorber has been proven over multiple generations of collider detectors. Recent detectors including UA1, CDF, and ZEUS have shown good results from such calorimeters. The advantages offered by scintillator calorimetry for the SSC environment, in particular, are speed (<10 nsec), excellent energy resolution, low noise, and ease of achieving compensation and hence linearity. On the negative side of the ledger can be placed the historical sensitivity of plastic scintillators to radiation damage, the possibility of nonuniform response because of light attenuation, and the presence of cracks for light collection via wavelength shifting plastic (traditionally in sheet form). This approach to calorimetry is being investigated for SSC use by a collaboration of Ames Laboratory/Iowa State University, Argonne National Laboratory, Bicron Corporation, Florida State University, Louisiana State University, University of Mississippi, Oak Ridge National Laboratory, Virginia Polytechnic Institute and State University, Westinghouse Electric Corporation, and University of Wisconsin.

  7. Intermittent plate tectonics?

    PubMed

    Silver, Paul G; Behn, Mark D

    2008-01-04

    Although it is commonly assumed that subduction has operated continuously on Earth without interruption, subduction zones are routinely terminated by ocean closure and supercontinent assembly. Under certain circumstances, this could lead to a dramatic loss of subduction, globally. Closure of a Pacific-type basin, for example, would eliminate most subduction, unless this loss were compensated for by comparable subduction initiation elsewhere. Given the evidence for Pacific-type closure in Earth's past, the absence of a direct mechanism for termination/initiation compensation, and recent data supporting a minimum in subduction flux in the Mesoproterozoic, we hypothesize that dramatic reductions or temporary cessations of subduction have occurred in Earth's history. Such deviations in the continuity of plate tectonics have important consequences for Earth's thermal and continental evolution.

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

    PubMed

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

    2010-02-01

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

  9. 10. DETAIL OF BUILDER'S PLATE AT NORTH PORTAL. PLATE READS: ...

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

    10. DETAIL OF BUILDER'S PLATE AT NORTH PORTAL. PLATE READS: 1889, BUILT BY THE BERLIN IRON BRIDGE CO. EAST BERLIN CONN. DOUGLAS & JARVIS PAT. APT. 16, 1878, AP'L 17, 1885. A.P. FORESMAN, WM. S. STARR, T.J. STREBEIGH, COMMISSIONERS. - Pine Creek Bridge, River Road spanning Pine Creek, Jersey Shore, Lycoming County, PA

  10. Downgoing plate controls on overriding plate deformation in subduction zones

    NASA Astrophysics Data System (ADS)

    Garel, Fanny; Davies, Rhodri; Goes, Saskia; Davies, Huw; Kramer, Stephan; Wilson, Cian

    2014-05-01

    Although subduction zones are convergent margins, deformation in the upper plate can be extensional or compressional and tends to change through time, sometimes in repeated episodes of strong deformation, e.g, phases of back-arc extension. It is not well understood what factors control this upper plate deformation. We use the code Fluidity, which uses an adaptive mesh and a free-surface formulation, to model a two-plate subduction system in 2-D. The model includes a composite temperature- and stress-dependent rheology, and plates are decoupled by a weak layer, which allows for free trench motion. We investigate the evolution of the state of stress and topography of the overriding plate during the different phases of the subduction process: onset of subduction, free-fall sinking in the upper mantle and interaction of the slab with the transition zone, here represented by a viscosity contrast between upper and lower mantle. We focus on (i) how overriding plate deformation varies with subducting plate age; (ii) how spontaneous and episodic back-arc spreading develops for some subduction settings; (iii) the correlation between overriding plate deformation and slab interaction with the transition zone; (iv) whether these trends resemble observations on Earth.

  11. Plate tectonics conserves angular momentum

    NASA Astrophysics Data System (ADS)

    Bowin, C.

    2010-03-01

    A new combined understanding of plate tectonics, Earth internal structure, and the role of impulse in deformation of the Earth's crust is presented. Plate accelerations and decelerations have been revealed by iterative filtering of the quaternion history for the Euler poles that define absolute plate motion history for the past 68 million years, and provide an unprecedented precision for plate angular rotation variations with time at 2-million year intervals. Stage poles represent the angular rotation of a plate's motion between adjacent Euler poles, and from which the maximum velocity vector for a plate can be determined. The consistent maximum velocity variations, in turn, yield consistent estimates of plate accelerations and decelerations. The fact that the Pacific plate was shown to accelerate and decelerate, implied that conservation of plate tectonic angular momentum must be globally conserved, and that is confirmed by the results shown here (total angular momentum ~1.4+27 kg m2 s-1). Accordingly, if a plate decelerates, other plates must increase their angular momentums to compensate. In addition, the azimuth of the maximum velocity vectors yields clues as to why the "bend" in the Emperor-Hawaiian seamount trend occurred near 46 Myr. This report summarizes processing results for 12 of the 14 major tectonic plates of the Earth (except for the Juan de Fuca and Philippine plates). Plate accelerations support the contention that plate tectonics is a product of torques that most likely are sustained by the sinking of positive density anomalies revealed by geoid anomalies of the degree 4-10 packet of the Earth's spherical harmonic coefficients. These linear positive geoid anomalies underlie plate subduction zones and are presumed due to phase changes in subducted gabbroic lithosphere at depth in the upper lower mantle (above 1200 km depth). The tectonic plates are pulled along by the sinking of these positive mass anomalies, rather than moving at near constant

  12. Aurelia aurita (Cnidaria) Oocytes' Contact Plate Structure and Development

    PubMed Central

    Adonin, Leonid S.; Shaposhnikova, Tatyana G.; Podgornaya, Olga

    2012-01-01

    One of the A. aurita medusa main mesoglea polypeptides, mesoglein, has been described previously. Mesoglein belongs to ZP-domain protein family and therefore we focused on A.aurita oogenesis. Antibodies against mesoglein (AB RA47) stain the plate in the place where germinal epithelium contacts oocyte on the paraffin sections. According to its position, we named the structure found the “contact plate”. Our main instrument was AB against mesoglein. ZP-domain occupies about half of the whole amino acid sequence of the mesoglein. Immunoblot after SDS-PAGE and AU-PAGE reveals two charged and high Mr bands among the female gonad germinal epithelium polypeptides. One of the gonads' polypeptides Mr corresponds to that of mesogleal cells, the other ones' Mr is higher. The morphological description of contact plate formation is the subject of the current work. Two types of AB RA47 positive granules were observed during progressive oogenesis stages. Granules form the contact plate in mature oocyte. Contact plate of A.aurita oocyte marks its animal pole and resembles Zona Pellucida by the following features: (1) it attracts spermatozoids; (2) the material of the contact plate is synthesized by oocyte and stored in granules; (3) these granules and the contact plate itself contain ZP domain protein(s); (4) contact plate is an extracellular structure made up of fiber bundles similar to those of conventional Zona Pellucida. PMID:23185235

  13. Regenerate metal-plating baths to cut waste and save

    SciTech Connect

    1995-03-01

    During electrode-based metal plating of equipment components, the formation of an electrical field causes metal deposits to be thicker at edges and seams, and thinner on flat surfaces. And, electrode-based methods consume large amounts of energy. Electroless-nickel (EN) -- or autocatalytic -- plating systems were pioneered in the 1940s to solve these problems. EN plating produces a more uniform coating, irrespective of the complexity of the part, and it consumes less electricity, since to electric current is required during plating. Plating in an EN system results from a chemical reaction between nickel in the bath and the substrate of the equipment component. The downside of electroless plating, however, is the limited life of the nickel bath, and the large volume of metal waste produced by bath disposal. Ionsep Corp. (Wilmington, Del.) has developed an electrodialytic system that continuously reforms the EN plating baths, to give them longer life. Its patented system has been successfully laboratory tested in a 1-ft{sup 2} cell, and the firm recently won a $250,000 grant from the US Dept. of Energy (Washington, D.C.) and Environmental Protection Agency (Washington, D.C.), to design and engineer a commercial-scale version of the system.

  14. Use of the quorum sensing inhibitor furanone C-30 to interfere with biofilm formation by Streptococcus mutans and its luxS mutant strain.

    PubMed

    He, Zhiyan; Wang, Qian; Hu, Yuejian; Liang, Jingping; Jiang, Yuntao; Ma, Rui; Tang, Zisheng; Huang, Zhengwei

    2012-07-01

    Streptococcus mutans is recognised as a major aetiological agent of dental caries. One of its important virulence factors is its ability to form biofilms on tooth surfaces. The aim of this study was to evaluate the effects of the quorum sensing inhibitor furanone C-30 on biofilm formation by S. mutans and its luxS mutant strain. The effects of furanone C-30 on biofilms of both strains formed on 96-well microtitre plates at 37 °C were determined by a colorimetric technique (MTT assay). Different concentrations of furanone C-30 (0.0, 2.0 and 4.0 μg/mL) and different time points of biofilm formation (4, 14 and 24 h) were investigated. The structures and thickness of the biofilms were observed by confocal laser scanning microscopy (CLSM). Quorum sensing-related gene expression (ftf, smu630, brpA, gbpB, gtfB, vicR, comDE and relA) was investigated by real-time polymerase chain reaction (RT-PCR). The results showed that synthetic furanone C-30 can inhibit biofilm formation by S. mutans and its luxS mutant strain, although it does not affect the bacterial growth rate itself. The quantities of biofilm formed by both strains significantly decreased (P<0.05) and the biofilms became thinner and looser as revealed by CLSM with increasing concentrations of furanone C-30. Expression of the genes tested was downregulated in the biofilms by the addition of furanone C-30. These results revealed that synthetic furanone C-30 can effectively inhibit biofilm formation by S. mutans and its luxS mutant strain.

  15. Plate tectonics, damage and inheritance.

    PubMed

    Bercovici, David; Ricard, Yanick

    2014-04-24

    The initiation of plate tectonics on Earth is a critical event in our planet's history. The time lag between the first proto-subduction (about 4 billion years ago) and global tectonics (approximately 3 billion years ago) suggests that plates and plate boundaries became widespread over a period of 1 billion years. The reason for this time lag is unknown but fundamental to understanding the origin of plate tectonics. Here we suggest that when sufficient lithospheric damage (which promotes shear localization and long-lived weak zones) combines with transient mantle flow and migrating proto-subduction, it leads to the accumulation of weak plate boundaries and eventually to fully formed tectonic plates driven by subduction alone. We simulate this process using a grain evolution and damage mechanism with a composite rheology (which is compatible with field and laboratory observations of polycrystalline rocks), coupled to an idealized model of pressure-driven lithospheric flow in which a low-pressure zone is equivalent to the suction of convective downwellings. In the simplest case, for Earth-like conditions, a few successive rotations of the driving pressure field yield relic damaged weak zones that are inherited by the lithospheric flow to form a nearly perfect plate, with passive spreading and strike-slip margins that persist and localize further, even though flow is driven only by subduction. But for hotter surface conditions, such as those on Venus, accumulation and inheritance of damage is negligible; hence only subduction zones survive and plate tectonics does not spread, which corresponds to observations. After plates have developed, continued changes in driving forces, combined with inherited damage and weak zones, promote increased tectonic complexity, such as oblique subduction, strike-slip boundaries that are subparallel to plate motion, and spalling of minor plates.

  16. Electronic readout systems for microchannel plates

    NASA Technical Reports Server (NTRS)

    Timothy, J. G.

    1985-01-01

    The modes of operation of position-sensitive electronic readout systems which use high-gain microchannel plate (MCP) electron multipliers are described, and their performance characteristics, along with those of the MCP, are compared. Among the structures presented are the wedge-and-strip, Codacon, and multilayer coincidence-anode MAMA (Multimode Microchannel Array) arrays. Spatial resolution of 25 x 25 sq microns (coincidence anode arrays) is achieved with an array format of 256 x 1024 pixels. On the basis of the performance data it is concluded that the readout systems using only conducting electrodes offer the best performance characteristics.

  17. ADOPT: A tool for automatic detection of tectonic plates at the surface of convection models

    NASA Astrophysics Data System (ADS)

    Mallard, C.; Jacquet, B.; Coltice, N.

    2017-08-01

    Mantle convection models with plate-like behavior produce surface structures comparable to Earth's plate boundaries. However, analyzing those structures is a difficult task, since convection models produce, as on Earth, diffuse deformation and elusive plate boundaries. Therefore we present here and share a quantitative tool to identify plate boundaries and produce plate polygon layouts from results of numerical models of convection: Automatic Detection Of Plate Tectonics (ADOPT). This digital tool operates within the free open-source visualization software Paraview. It is based on image segmentation techniques to detect objects. The fundamental algorithm used in ADOPT is the watershed transform. We transform the output of convection models into a topographic map, the crest lines being the regions of deformation (plate boundaries) and the catchment basins being the plate interiors. We propose two generic protocols (the field and the distance methods) that we test against an independent visual detection of plate polygons. We show that ADOPT is effective to identify the smaller plates and to close plate polygons in areas where boundaries are diffuse or elusive. ADOPT allows the export of plate polygons in the standard OGR-GMT format for visualization, modification, and analysis under generic softwares like GMT or GPlates.

  18. Metals plated on fluorocarbon polymers

    NASA Technical Reports Server (NTRS)

    Ford, H.; Krasinsky, J. B.; Vango, S. P.

    1964-01-01

    Electroplating lead on fluorocarbon polymer parts is accomplished by etching the parts to be plated with sodium, followed by successive depositions of silver and lead from ultrasonically agitated plating solutions. Metals other than lead may be electroplated on the silvered parts.

  19. Metal vapor arc ion plating

    DOEpatents

    Bertram, L.A.; Fisher, R.W.; Mattox, D.M.; Zanner, F.J.

    1986-09-09

    A method and apparatus for ion plating are described. The apparatus uses more negative than a first electrode voltage in a vacuum arc remelt system to attract low energy ions from the anode electrode to the article to be plated. 2 figs.

  20. Micro-channel plate detector

    DOEpatents

    Elam, Jeffrey W.; Lee, Seon W.; Wang, Hsien -Hau; Pellin, Michael J.; Byrum, Karen; Frisch, Henry J.

    2015-09-22

    A method and system for providing a micro-channel plate detector. An anodized aluminum oxide membrane is provided and includes a plurality of nanopores which have an Al coating and a thin layer of an emissive oxide material responsive to incident radiation, thereby providing a plurality of radiation sensitive channels for the micro-channel plate detector.

  1. Gold plating on spectacle frames.

    PubMed

    Kenny, I; Mitchell, J W; Walsh, G

    1997-05-01

    An investigation was carried out into the thickness and standard of application of the plating and lacquer coatings applied to three metal spectacle frames. All conform to BS 6625 (1991) for plating thickness, but there was considerable variation in regularity and porosity.

  2. Delaminations in composite plates under impact loads

    NASA Technical Reports Server (NTRS)

    Finn, Scott R.; Springer, George S.

    1991-01-01

    A method is presented for calculating the locations, shapes, and sizes of delaminations which occur in a fiber reinforced composite plate subjected to non-penetrating (low velocity) impact of a solid object. The plate may be simply supported, clamped, or free along its edges. A failure model of the delamination formation was developed. This model was then coupled with a finite element analysis. The model and the finite element analysis were then implemented by a computer code (IMPACT-ST) which can be used to estimate the damage initiation load and the locations, shapes, and sizes of the delaminations. Tests were performed measuring the geometries of the delaminations in graphite-epoxy, graphite-toughened epoxy, and graphite-PEEK plates impacted by a projectile with a spherical tip having masses ranging from 0.355 lbm to 0.963 lbm and velocities from 50 in/sec to 225 in/sec. The data were compared to the results of the model, and good agreements were found between the measured and the calculated delamination lengths and widths.

  3. Plate Interface Rheology, Mechanical Coupling and Accretion during Subduction Infancy

    NASA Astrophysics Data System (ADS)

    Agard, P.; Yamato, P.; Mathieu, S.; Prigent, C.; Guillot, S.; Plunder, A.; Dubacq, B.; Monie, P.; Chauvet, A.

    2015-12-01

    Understanding subduction rheology in both space and time has been a challenge since the advent of plate tectonics. We herein focus on "subduction infancy", that is the first ~1-5 My immediately following subduction nucleation, when a newly born slab penetrates into the upper plate mantle and heats up. The only remnants of this critical yet elusive geodynamic step are thin metamorphic soles, commonly found beneath pristine, 100-1000 km long portions of oceanic lithosphere emplaced on continents (i.e., ophiolites). Through the (i) worldwide compilation of pressure-temperature conditions of metamorphic sole formation augmented by pseudosection thermodynamic modeling, (ii) calculations of the viscosity of materials along the plate interface and (iii) generic numerical thermal models, we provide a conceptual model of dynamic plate interface processes during subduction infancy (and initiation s.l.). We show in particular how major rheological switches across the subduction interface control slab penetration, and the formation of metamorphic soles. Due to the downward progression of hydration and weakening of the mantle wedge with cooling, the lower plate (basalt, sediment) and the upper plate (mantle wedge) rheologies equalize and switch over a restricted temperature-time-depth interval (e.g., at ~800°C and ~1 GPa, during 0.1-2 My, for high-temperature metamorphic sole formation). These switches result in episodes of maximum interplate mechanical coupling, thereby slicing the top of the slab and welding pieces (basalt, sediment) to the base of the mantle wedge. Similar mechanical processes likely apply for the later, deeper accretion and exhumation of high-temperature oceanic eclogites in serpentinite mélanges, or for the accretion of larger tectonic slices. This model provides constraints on the effective rheologies of the crust and mantle and general understanding, at both rock and plate scale, for accretion processes and early slab dynamics.

  4. Aseptic Laboratory Techniques: Plating Methods

    PubMed Central

    Sanders, Erin R.

    2012-01-01

    Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: ● Perform plating procedures without contaminating media. ● Isolate single bacterial colonies by the

  5. Aseptic laboratory techniques: plating methods.

    PubMed

    Sanders, Erin R

    2012-05-11

    Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: Perform plating procedures without contaminating media. Isolate single bacterial colonies by the streak-plating

  6. Strong Plate, Weak Slab Dichotomy

    NASA Astrophysics Data System (ADS)

    Petersen, R. I.; Stegman, D. R.; Tackley, P.

    2015-12-01

    Models of mantle convection on Earth produce styles of convection that are not observed on Earth.Moreover non-Earth-like modes, such as two-sided downwellings, are the de facto mode of convection in such models.To recreate Earth style subduction, i.e. one-sided asymmetric recycling of the lithosphere, proper treatment of the plates and plate interface are required. Previous work has identified several model features that promote subduction. A free surface or pseudo-free surface and a layer of material with a relatively low strength material (weak crust) allow downgoing plates to bend and slide past overriding without creating undue stress at the plate interface. (Crameri, et al. 2012, GRL)A low viscosity mantle wedge, possibly a result of slab dehydration, decouples the plates in the system. (Gerya et al. 2007, Geo)Plates must be composed of material which, in the case of the overriding plate, are is strong enough to resist bending stresses imposed by the subducting plate and yet, as in the case of the subducting plate, be weak enough to bend and subduct when pulled by the already subducted slab. (Petersen et al. 2015, PEPI) Though strong surface plates are required for subduction such plates may present a problem when they encounter the lower mantle.As the subducting slab approaches the higher viscosity, lower mantle stresses are imposed on the tip.Strong slabs transmit this stress to the surface.There the stress field at the plate interface is modified and potentially modifies the style of convection. In addition to modifying the stress at the plate interface, the strength of the slab affects the morphology of the slab at the base of the upper mantle. (Stegman, et al 2010, Tectonophysics)Slabs that maintain a sufficient portion of their strength after being bent require high stresses to unbend or otherwise change their shape.On the other hand slabs that are weakened though the bending process are more amenable to changes in morphology. We present the results of

  7. Miniaturization of a homogeneous fluorescence immunoassay based on energy transfer using nanotiter plates as high-density sample carriers.

    PubMed

    Schobel, U; Coille, I; Brecht, A; Steinwand, G M; Gauglitz, G

    2001-11-01

    The miniaturization of a homogeneous competitive immunoassay to a final assay volume of 70 nL is described. As the sample carrier, disposable plastic nanotiter plates (NTP) with dimensions of 2 x 2 cm2 containing 25 x 25 wells, corresponding to approximately 15,000 wells on a traditional 96-well microtiter plate footprint, were used. Sample handling was accomplished by a piezoelectrically actuated micropipet. To reduce evaporation while pipetting the assays, the NTP was handled in a closed humid chamber and cooled to the point of condensation. To avoid washing steps, a homogeneous assay was developed that was based on energy-transfer (ET). As a model system, an antibody-based assay for the detection of the environmentally relevant compound, simazine, in drinking water was chosen. Antibodies were labeled with the long-wavelength-excitable sulfoindocyanine dye Cy5 (donor), and a tracer was synthesized by labeling BSA with a triazine derivative and the acceptor dye Cy5.5. At low analyte concentrations, the tracer was preferably bound to the antibody binding sites. As a result of the close proximity of Cy5.5 and Cy5, an efficient quenching of the Cy5 fluorescence occurred. Higher analyte concentrations led to a progressive binding of the analyte to the antibody binding sites. The increased Cy5 fluorescence was determined by using a scanning laser-induced fluorescence detector. The limit of detection (LOD), using an antibody concentration of 20 nM, was 0.32 microg/L, or 1.11 x 10(-16) mol of simazine. In comparison, the LOD of the 96-well microtiter-plate-based ET immunoassay (micro-ETIA) was 0.15 microg/L, or 1.87 x 10(-13) mol. The LOD of the optimized micro-ETIA at 1 nM IgG, was 0.01 microg/L.

  8. Present-day plate motions

    NASA Technical Reports Server (NTRS)

    Minster, J. B.; Jordan, T. H.

    1977-01-01

    A data set comprising 110 spreading rates, 78 transform fault azimuths and 142 earthquake slip vectors was inverted to yield a new instantaneous plate motion model, designated RM2. The mean averaging interval for the relative motion data was reduced to less than 3 My. A detailed comparison of RM2 with angular velocity vectors which best fit the data along individual plate boundaries indicates that RM2 performs close to optimally in most regions, with several notable exceptions. On the other hand, a previous estimate (RM1) failed to satisfy an extensive set of new data collected in the South Atlantic Ocean. It is shown that RM1 incorrectly predicts the plate kinematics in the South Atlantic because the presently available data are inconsistent with the plate geometry assumed in deriving RM1. It is demonstrated that this inconsistency can be remedied by postulating the existence of internal deformation with the Indian plate, although alternate explanations are possible.

  9. The moving plate capacitor paradox

    NASA Astrophysics Data System (ADS)

    Davis, B. R.; Abbott, D.; Parrondo, J. M. R.

    2000-03-01

    For the first time we describe an apparent paradox concerning a moving plate capacitor driven by thermal noise from a resistor. A demon restores the plates of the capacitor to their original position, only when the voltage across the capacitor is small—hence only small forces are present for the demon to work against. The demon has to work harder than this to avoid the situation of perpetual motion, but the question is how? We explore the concept of a moving plate capacitor, driven by noise, a step further by examining the case where the restoring force on the capacitor plates is provided by a simple spring, rather than some unknown demon. We display simulation results with interesting behavior, particularly where the capacitor plates collide with each other.

  10. Sub-Plate Overlap Code Documentation

    NASA Technical Reports Server (NTRS)

    Taff, L. G.; Bucciarelli, B.; Zarate, N.

    1997-01-01

    An expansion of the plate overlap method of astrometric data reduction to a single plate has been proposed and successfully tested. Each plate is (artificially) divided into sub-plates which can then be overlapped. This reduces the area of a 'plate' over which a plate model needs to accurately represent the relationship between measured coordinates and standard coordinates. Application is made to non-astrographic plates such as Schmidt plates and to wide-field astrographic plates. Indeed, the method is completely general and can be applied to any type of recording media.

  11. Statistical tests of additional plate boundaries from plate motion inversions

    NASA Technical Reports Server (NTRS)

    Stein, S.; Gordon, R. G.

    1984-01-01

    The application of the F-ratio test, a standard statistical technique, to the results of relative plate motion inversions has been investigated. The method tests whether the improvement in fit of the model to the data resulting from the addition of another plate to the model is greater than that expected purely by chance. This approach appears to be useful in determining whether additional plate boundaries are justified. Previous results have been confirmed favoring separate North American and South American plates with a boundary located beween 30 N and the equator. Using Chase's global relative motion data, it is shown that in addition to separate West African and Somalian plates, separate West Indian and Australian plates, with a best-fitting boundary between 70 E and 90 E, can be resolved. These results are generally consistent with the observation that the Indian plate's internal deformation extends somewhat westward of the Ninetyeast Ridge. The relative motion pole is similar to Minster and Jordan's and predicts the NW-SE compression observed in earthquake mechanisms near the Ninetyeast Ridge.

  12. Underwater electrical discharge in plate to plate configuration

    NASA Astrophysics Data System (ADS)

    Stelmashuk, Vitaliy

    2016-09-01

    Two main configurations of high voltage electrodes submersed in water have been used for an electrical discharge generation: pin to pin and pin to plate. An electrical breakdown between plate electrodes is generally difficult to reproduce, because there is a uniform and weak electric field. One major advantage of using plate electrodes is their greater ``wear hardness'' to high-energy discharges. The plate electrodes can withstand extremely high energy deposition at which the pin electrode is quickly destroyed. The electrical discharge between plate electrodes can be initiated by creating an inhomogeneity in the electrical field. Two methods of discharge initiation between plate electrodes are proposed for this aim: 1) focusing of a shock wave in the interelectrode region; 2) a bubble injection into the electrode gap. The shock wave creates favourable conditions for the electrical breakdown between the two plate electrodes: it causes that numerous microbubbles of dissolved air start to grow and serve as locations for streamer initiation. In the second method the gas bubble is injected from the one of the electrodes, which has a gas inlet hole on the lateral face for this purpose. A ``volcano'' like morphology of positive streamers are observed in the experiments with weak electric field. The authors are grateful to MEYS grant INGO LG 15013.

  13. Statistical tests of additional plate boundaries from plate motion inversions

    NASA Technical Reports Server (NTRS)

    Stein, S.; Gordon, R. G.

    1984-01-01

    The application of the F-ratio test, a standard statistical technique, to the results of relative plate motion inversions has been investigated. The method tests whether the improvement in fit of the model to the data resulting from the addition of another plate to the model is greater than that expected purely by chance. This approach appears to be useful in determining whether additional plate boundaries are justified. Previous results have been confirmed favoring separate North American and South American plates with a boundary located beween 30 N and the equator. Using Chase's global relative motion data, it is shown that in addition to separate West African and Somalian plates, separate West Indian and Australian plates, with a best-fitting boundary between 70 E and 90 E, can be resolved. These results are generally consistent with the observation that the Indian plate's internal deformation extends somewhat westward of the Ninetyeast Ridge. The relative motion pole is similar to Minster and Jordan's and predicts the NW-SE compression observed in earthquake mechanisms near the Ninetyeast Ridge.

  14. Thin plate neotectonic models of the Australian plate

    NASA Astrophysics Data System (ADS)

    Burbidge, D. R.

    2004-10-01

    Thin plate finite element models of the neotectonic deformation of the Australian plate have been calculated in order to estimate the stress and strain rate within the plate, specifically concentrating on the Australian continent. The model includes plate-bounding faults, an anelastic brittle-ductile layered rheology and the option of laterally varying elevation and heat flow. The results of the models are compared to (1) the velocity of geodetic benchmarks on the Australian plate, (2) the spreading rate of the mid-oceanic ridges along the Australian plate's margins, (3) the direction of the maximum horizontal principal stress, (4) the stress regime within the plate, and (5) the crustal thickness estimated from the depth to the base of Mohorovicic discontinuity's transition zone. A variety of models are tested with a wide range of input parameters. The model with the smallest misfit with observations predicts that the strain rate for most of the Australian continent is approximately 10-17 s-1. This model has a slightly lower strain rate in the central Australia and is higher off the northern coast of Australia than for the rest of the continent. Strain rates of this magnitude would be difficult to observe from geodetic or geologic data for most parts of Australia but would be enough to generate much of the seismicity that has been observed over the last century.

  15. Highly curved microchannel plates

    NASA Technical Reports Server (NTRS)

    Siegmund, O. H. W.; Cully, S.; Warren, J.; Gaines, G. A.; Priedhorsky, W.; Bloch, J.

    1990-01-01

    Several spherically curved microchannel plate (MCP) stack configurations were studied as part of an ongoing astrophysical detector development program, and as part of the development of the ALEXIS satellite payload. MCP pairs with surface radii of curvature as small as 7 cm, and diameters up to 46 mm have been evaluated. The experiments show that the gain (greater than 1.5 x 10 exp 7) and background characteristics (about 0.5 events/sq cm per sec) of highly curved MCP stacks are in general equivalent to the performance achieved with flat MCP stacks of similar configuration. However, gain variations across the curved MCP's due to variations in the channel length to diameter ratio are observed. The overall pulse height distribution of a highly curved surface MCP stack (greater than 50 percent FWHM) is thus broader than its flat counterpart (less than 30 percent). Preconditioning of curved MCP stacks gives comparable results to flat MCP stacks, but it also decreases the overall gain variations. Flat fields of curved MCP stacks have the same general characteristics as flat MCP stacks.

  16. Highly curved microchannel plates

    NASA Technical Reports Server (NTRS)

    Siegmund, O. H. W.; Cully, S.; Warren, J.; Gaines, G. A.; Priedhorsky, W.; Bloch, J.

    1990-01-01

    Several spherically curved microchannel plate (MCP) stack configurations were studied as part of an ongoing astrophysical detector development program, and as part of the development of the ALEXIS satellite payload. MCP pairs with surface radii of curvature as small as 7 cm, and diameters up to 46 mm have been evaluated. The experiments show that the gain (greater than 1.5 x 10 exp 7) and background characteristics (about 0.5 events/sq cm per sec) of highly curved MCP stacks are in general equivalent to the performance achieved with flat MCP stacks of similar configuration. However, gain variations across the curved MCP's due to variations in the channel length to diameter ratio are observed. The overall pulse height distribution of a highly curved surface MCP stack (greater than 50 percent FWHM) is thus broader than its flat counterpart (less than 30 percent). Preconditioning of curved MCP stacks gives comparable results to flat MCP stacks, but it also decreases the overall gain variations. Flat fields of curved MCP stacks have the same general characteristics as flat MCP stacks.

  17. Balanced Orifice Plate

    NASA Technical Reports Server (NTRS)

    Kelley, Anthony R. (Inventor); Buskirk, Paul D. (Inventor)

    2006-01-01

    An orifice plate for use in a conduit through which fluid flows is defined by a central circular region having a radius R, and a ring-shaped region surrounding the central circular region. The ring-shaped region has holes formed therethrough with those holes centered at each radius R thereof satisfying a relationship A(sub R)=al(X(sub R)V(sub R)(sup b)) where A(sub R) is a sum of areas of those holes having centers at radius R, X(sub R) is a flow coefficient at radius R, V(sub R) is a velocity of the fluid that is to flow through the conduit at radius R, b is a constant selected to make at least one process variable (associated with the fluid that is to flow through the conduit) approximately equal at each radius R, and a is a constant that is equal to (X(sub R)A(sub R)V(sub R)(sup b)) at each radius R.

  18. Evaluation of a non-cyanide gold plating process for switch tubes

    SciTech Connect

    Norwood, D.P.; Martinez, F.E.

    1996-01-01

    Switch tubes are used in nuclear weapon firing sets and are required to be reliable and impervious to gas permeation for many years. To accomplish this, a gold plated coating of approximately 25 microns is required over all metal surfaces on the tube exterior. The gold has historically been plated using gold cyanide plating chemistry. In this work we proposed to replace the cyanide plating bath with an environmentally friendlier sulfite gold plating bath. Low and high pH sulfite plating chemistries were investigated as possible replacements for the cyanide gold plating chemistry. The low pH plating chemistry demonstrated a gold plated coating which met the high purity, grain size, and hardness requirements for switch tubes. The high pH chemistry was rejected primarily because the hardness of the gold plated coatings was too high and exceeded switch tube coating requirements. A problem with nodule formation on the gold plated surface using the low pH chemistry had to be resolved during this evaluation. The nodule formation was postulated to be produced by generation of SO{sub 2} in the low pH bath causing gold to be precipitated out when the sulfite concentration falls below a minimum level. The problem was resolved by maintaining a higher sulfite concentration and providing an active filtration system during plating. In this initial study, there were no major obstacles found when using a sulfite gold bath for switch tube plating, however, further work is needed on bath control and bath life before adopting it as the primary plating chemistry.

  19. Neural crest induction at the neural plate border in vertebrates.

    PubMed

    Milet, Cécile; Monsoro-Burq, Anne H

    2012-06-01

    The neural crest is a transient and multipotent cell population arising at the edge of the neural plate in vertebrates. Recent findings highlight that neural crest patterning is initiated during gastrulation, i.e. earlier than classically described, in a progenitor domain named the neural border. This chapter reviews the dynamic and complex molecular interactions underlying neural border formation and neural crest emergence.

  20. Impact of the overriding plate rheology on convergence zone dynamics.

    NASA Astrophysics Data System (ADS)

    Hertgen, Solenn; Yamato, Philippe; Guillaume, Benjamin; van Hunen, Jeroen

    2017-04-01

    Most of deformation at the Earth's surface is localized at plate boundaries. This deformation can be accommodated in very different ways depending on the tectonic setting. In the case of convergence zones, the deformation is typically simplified and classified as follows: - intra-oceanic convergence, when convergence involves two oceanic lithospheres, which generally leads to the subduction/obduction initiation and to the formation of an island arc; - convergence between an oceanic and a continental lithosphere, which is mainly accommodated by subduction and can lead to the formation of a mountain range at the plate boundary; - convergence involving two continental lithospheres, which is accommodated by collision and leads to the formation of a mountain range produced by the stacking of crustal slices. Different materials are thus involved (i.e., oceanic crust, continental crust, sediments). Depending on the context (oceanic or continental subduction), they can form contrasted structures in terms of units size, morphology and metamorphism (e.g., Alps vs. Andes/Altiplano-Puna). Moreover, some convergent zones with apparently similar tectonic settings (e.g., continent/continent convergence) show very different patterns of deformation with either very localized deformation (e.g., the Alps) or, at the opposite, deformation distributed over thousands of kilometers (e.g, Himalayas/Tibet). Finally, other convergent zones from different tectonic settings seem to show similar structures (e.g., Tibet plateau and Altiplano-Puna plateaus). Although the mechanism of plate convergence appears to be the same in each case, the structures obtained at the surface seem to be unique. Rheology of both the subducting plate and of the plate interface is known to influence the convergence zones dynamics. However, very few studies have addressed the role of the overriding plate rheology in details, while it may also exert a large control on the deformation style at plate boundaries. In

  1. Biomechanics of bone-fracture fixation by stiffness-graded plates in comparison with stainless-steel plates

    PubMed Central

    Ganesh, VK; Ramakrishna, K; Ghista, Dhanjoo N

    2005-01-01

    Background In the internal fixation of fractured bone by means of bone-plates fastened to the bone on its tensile surface, an on-going concern has been the excessive stress-shielding of the bone by the excessively-stiff stainless-steel plate. The compressive stress-shielding at the fracture-interface immediately after fracture-fixation delays callus formation and bone healing. Likewise, the tensile stress-shielding of the layer of the bone underneath the plate can cause osteoporosis and decrease in tensile strength of this layer. Method In order to address this problem, we propose to use stiffness-graded plates. Accordingly, we have computed (by finite-element analysis) the stress distribution in the fractured bone fixed by composite plates, whose stiffness is graded both longitudinally and transversely. Results It can be seen that the stiffness-graded composite-plates cause less stress-shielding (as an example: at 50% of the healing stage, stress at the fracture interface is compressive in nature i.e. 0.002 GPa for stainless steel plate whereas stiffness graded plates provides tensile stress of 0.002 GPa. This means that stiffness graded plate is allowing the 50% healed bone to participate in loadings). Stiffness-graded plates are more flexible, and hence permit more bending of the fractured bone. This results in higher compressive stresses induced at the fractured faces accelerate bone-healing. On the other hand, away from the fracture interface the reduced stiffness and elastic modulus of the plate causes the neutral axis of the composite structure to be lowered into the bone resulting in the higher tensile stress in the bone-layer underneath the plate, wherein is conducive to the bone preserving its tensile strength. Conclusion Stiffness graded plates (with in-built variable stiffness) are deemed to offer less stress-shielding to the bone, providing higher compressive stress at the fractured interface (to induce accelerated healing) as well as higher tensile

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

    PubMed

    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

  3. 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

  4. Tectonic Plates of China

    DTIC Science & Technology

    1977-04-01

    continental crust formed before the Sinian time. They were later more or less covered by strata of different periods , metamorphozed and folded (in case of...tectonic cycles and ten orogenic periods in the formation of the fault blocks of China. Their names and time intervals, together with the blocks formed...during each period , are listed in Table 1. The positions of the blocks are shown in Fig. 1. (3) The fault blocks are separated by fault zones. Three types

  5. Buckling analysis of laminated plates using partial hybrid plate element

    SciTech Connect

    Tseng, Y.P.; Chou, L.C.

    1995-08-01

    In this paper, the stability of laminated composite plates was studied by a partial hybrid plate element based on higher order plate theory. The transverse shear deformation and the warping of cross section are both considered by the higher order plate theory. Moreover, the element stiffness is formulated by only assuming transverse shear stress field, while the flexural stresses from assumed displacement field. Consequently, the interlaminar transverse shear discontinuity is overcome. On the other hand, the buckling load is assumed to be uniform through thickness in-plane stresses in this study. The Green-Lagrangian strains, instead of Von Karmon strains, are then adopted to take the nonlinear in-plane deformations into account. Numerical results are presented for several benchmark problems including various span-to-thickness ratios, stacking sequences, boundary conditions, and loading cases.

  6. Plate interactions of Laurussia and Gondwana during the formation of Pangea- Constraints by U-Pb LA-SF-ICP-MS detrital zircon ages of Devonian and Early Carboniferous siliciclastics of the Rheinisches Schiefergebirge (Rhenohercynian zone, Central European

    NASA Astrophysics Data System (ADS)

    Eckelmann, Katja; Nesbor, Heinz-Dieter; Königshof, Peter; Linnemann, Ulf; Hofmann, Mandy; Lange, Jan-Michael; Sagawe, Anja

    2013-04-01

    The Rhenohercynian zone of the Central European Variscides contains top-NW thrusted rock complexes in the area of the Hörre nappe. Further to the north, par-authochthonous volcano-sedimentary complexes of the Lahn and Dill-Eder synclines occur. In our paper, a representative data set of U-Pb LA-SF ICP-MS ages of 1228 detrital zircon grains of Devonian and Lower Carboniferous sandstones from both areas is presented. The data constrain the Middle to Upper Devonian and the Lower Carboniferous siliciclastic and volcano-sedimentary complexes of the Hörre nappe to be allochtonous. The cluster of U-Pb ages of detrital zircon grains point to a provenance of the debris from the Saxo-Thuringian zone and rock complexes of the Mid-German Crystalline zone, which were formerly related to the latter one. A long-distance top-NW directed transport of the nappe complex in the Hörre nappe is required by our new data set. The zircon populations derived from the Saxo-Thuringian zone are representative for a Gondwanan hinterland and characterised by age clusters of ~530-700 Ma, ~1,8-2.2 Ga, ~2.5-2.7 Ga, and ~3.0-3.4 Ga. In a palaeogeographic view, the Saxo-Thuringian zone formed during Devonian to Lower Carboniferous time at the southern margin of the Rheic Ocean. A Lower Devonian sandstone sample from the par-authochtonous Lahn-Dill-Eder syncline shows a provenance, which is representative for Laurussia. Most of that debris seems to be derived from Baltica and Avalonia, two important parts of Laurussia. U-Pb zircon ages cluster at ~ 400-450 Ma, 540-650 Ma, 1.0-1.2 Ga, ~1.4-1.5 Ga, ~1.7-2.2 Ga, and 2.3-2.9 Ga. In terms of a palaeogeographic relation, these samples represent a sedimentary unit which was deposited at the northern margin of the Rheic Ocean at the edge of Laurussia facing to the south. Our data set constrains a top-SE directed subduction of the oceanic crust of the Rheic Ocean during its final closure and the formation of supercontinent Pangea.

  7. High loading uranium fuel plate

    DOEpatents

    Wiencek, Thomas C.; Domagala, Robert F.; Thresh, Henry R.

    1990-01-01

    Two embodiments of a high uranium fuel plate are disclosed which contain a meat comprising structured uranium compound confined between a pair of diffusion bonded ductile metal cladding plates uniformly covering the meat, the meat having a uniform high fuel loading comprising a content of uranium compound greater than about 45 Vol. % at a porosity not greater than about 10 Vol. %. In a first embodiment, the meat is a plurality of parallel wires of uranium compound. In a second embodiment, the meat is a dispersion compact containing uranium compound. The fuel plates are fabricated by a hot isostatic pressing process.

  8. Plating on stainless steel alloys

    SciTech Connect

    Dini, J.W.; Johnson, H.R.

    1981-09-11

    Quantitative adhesion data are presented for a variety of electroplated stainless steel type alloys. Results show that excellent adhesion can be obtained by using a Wood's nickel strike or a sulfamate nickel strike prior to final plating. Specimens plated after Wood's nickel striking failed in the deposit rather than at the interface between the substrate and the coating. Flyer plate quantitative tests showed that use of anodic treatment in sulfuric acid prior to Wood's nickel striking even further improved adhesion. In contrast activation of stainless steels by immersion or cathodic treatment in hydrochloric acid resulted in very reduced bond strengths with failure always occurring at the interface between the coating and substrate.

  9. D-Amino Acids Enhance the Activity of Antimicrobials against Biofilms of Clinical Wound Isolates of Staphylococcus aureus and Pseudomonas aeruginosa

    DTIC Science & Technology

    2014-05-19

    96-well plates and biofilm dispersal assays. Biofilm formation was performed under static conditions for 24 h in polystyrene 96-well plates (Corning...each well for an additional 24 h . Following overnight exposure, cells were washed as above and biofilm biomass was determined by measuring the optical...pegs for the attachment of the bacteria, and incubated at 37°C for 48 h with agitation. Following incubation, plate lids containing the pegs with the

  10. Origin of plate tectonics: Grain-damage, inheritance and hysteresis

    NASA Astrophysics Data System (ADS)

    Bercovici, D.; Ricard, Y. R.

    2015-12-01

    The emergence of plate tectonics is enigmatic because of the lack of observations in the early Archean as well as the challenge of understanding how plates form. The damage theory of lithospheric weakening by grain-reduction provides a physical framework for plate generation. This model builds on grain-scale physics to describe planetary-scale processes, and is consistent with lab and field observations of polycrystalline rocks and lithospheric mylonites. Grain-damage accounts for the evolution of damage and healing by grain growth, hence predicts plate boundary formation and longevity, and how they depend on surface conditions. The establishment of global plate tectonics likely started between >4Ga and 2.7Ga, and may have taken over a billion years to develop. Under Earth-like conditions, grain-damage combined with intermittent Archean protosubduction produces persistent weak zones that accumulate into well developed plates by 3Ga. However, Venus' hotter surface promotes healing, suppresses damage and inhibits weak zone accumulation, which suggests why plate tectonics failed to spread on our sister planet. New work posits that interface damage is possibly suppressed at moderate grain-size; this induces a hysteresis loop wherein three equilibrium deformation branches coexist. These branches include a stable large-grain, weakly-deforming state in dislocation creep, a stable small-grain rapidly-deforming state in diffusion creep analogous to mylonites, and an unstable intermediate-grain state. At the right conditions, a lithosphere can acquire two stable deformation states characteristic of plate tectonics; i.e., both slowly deforming plate interiors and rapidly deforming plate boundaries can co-exist. Earth currently sits inside the hysteresis loop and can have coexisting deformation states, while Venus sits at the end of the loop where only the weakly deforming branch dominates. The hot post-Hadean Earth might have had peak deformation only on the weakly

  11. Preparation and magnetic properties of Ni-P-La coating by electroless plating on silicon substrate

    NASA Astrophysics Data System (ADS)

    Gao, Yun; Wang, Jihui; Yuan, Jing; Li, Haiqin

    2016-02-01

    Ni-P-La coatings were prepared on Si substrate by electroless plating method under different La content, pH value, plating temperature and plating time. The surface morphology, chemical composition, structure and magnetic properties of coatings were observed and determined by scanning electron microscope (SEM), energy dispersive X-ray spectrometry (EDS), X-ray diffractometer (XRD) and vibrating sample magnetometer (VSM). The results showed that Ni-P-La coating is smooth and uniform with a cellular morphology grown in columnar manner. With the increase of La content, pH value and plating time, the thickness and saturation magnetization of coating are increased continuously, but the stability of plating bath is decreased greatly with La content and pH value. Under higher plating temperature, the thickness and saturation magnetization of coatings are obviously enhanced. But too high plating temperature is harmful to the plating bath and coating. The optimum plating conditions for Ni-P-La coating is La2O3 addition of 10 mg L-1, pH value of 5.0, plating temperature of 75 °C and plating time of 45 min. The role of La element is to benefit the deposition of Ni element, promote the formation of Ni phase during the annealing process, and thus improve the magnetic properties of Ni-P-La coating.

  12. Evaluation of prominence of straight plates and precontoured clavicle plates using automated plate-to-bone alignment.

    PubMed

    Van Tongel, A; Huysmans, T; Amit, B; Sijbers, J; Vanglabbeek, F; De Wilde, L

    2014-09-01

    Hardware prominence after plate fixation for clavicle fracture is a common complication. The aim of the study was to perform a 3D analysis of the prominence of different types of superior clavicle plates. An automated fitting of 3 straight and 10 precontoured plates was performed on 52 3D-CT-scan reconstructed cadaver clavicles. The mean and maximum bone-plate distance and maximum prominence was significant higher with the straight plates compared to the precontoured plates. The mean and maximum boneplate distance was significant higher with the precontoured DePuy-Synthes plates compared to the precontoured Acumed plates but when evaluating the maximum prominence there was no significant difference between the most commonly used 8-holes plates. To conclude, precontoured plates of the clavicula diminish significantly hardware prominence. There exists a difference in hardware prominence between different brands of precontoured plates but this difference is limited and in most cases not significant.

  13. Spot brazing of aluminum to copper with a cover plate

    NASA Astrophysics Data System (ADS)

    Hayashi, Junya; Miyazawa, Yasuyuki

    2014-08-01

    It is difficult to join dissimilar metals when an intermetallic compound is formed at the joining interface. Spot brazing can be accomplished in a short time by resistance heating. Therefore, it is said that the formation of a intermetallic compound can be prevented. In this study, aluminum and copper were joined by spot brazing with a cover plate. The cover plate was used to supply heat to base metals and prevent heat dissipation from the base metals. The ability to braze Al and Cu was investigated by observation and analysis. Pure aluminum (A1050) plate and oxygen-free copper (C1020) plate were used as base metals. Cu-Ni-Sn-P brazing filler was used as the brazing filler metal. SPCC was employed as cover plate. Brazing was done with a micro spot welder under an argon gas atmosphere. Brazing ability was estimated by tensile shear strength and cross sectional microstructure observation. Al and Cu can be joined by spot brazing with Cu-Ni-Sn-P brazing filler and cover plate.

  14. The multigap resistive plate chamber

    SciTech Connect

    Zeballos, E. Cerron; Crotty, I.; Hatzifotiadou, D.; Valverde, J. Lamas; Neupane, S.; Williams, M. C. S.; Zichichi, A.

    2015-02-03

    The paper describes the multigap resistive plate chamber (RPC). This is a variant of the wide gap RPC. However it has much improved time resolution, while keeping all the other advantages of the wide gap RPC design.

  15. Carbon-assisted flyer plates

    DOEpatents

    Stahl, D.B.; Paisley, D.L.

    1994-04-12

    A laser driven flyer plate is described utilizing an optical fiber connected to a laser. The end of the optical fiber has a layer of carbon and a metal layer deposited onto it. The carbon layer provides the laser induced plasma which is superior to the plasma produced from most metals. The carbon layer plasma is capable of providing a flatter flyer plate, converting more of the laser energy to driving plasma, promoting a higher flyer plate acceleration, and providing a more uniform pulse behind the plate. In another embodiment, the laser is in optical communication with a substrate onto which a layer of carbon and a layer of metal have been deposited. 2 figures.

  16. Tectonics: Changing of the plates

    NASA Astrophysics Data System (ADS)

    Brandon, Alan

    2016-10-01

    The composition of Earth's crust depends on the style of plate tectonics and of the melting regimes in the mantle. Analyses of the oldest identified rocks suggest that these styles and the resulting crust have changed over Earth's history.

  17. Reflection-Zone-Plate Antenna

    NASA Technical Reports Server (NTRS)

    Franke, John M.; Leighty, Bradley D.

    1989-01-01

    Microwave antenna, based on reflection holography, designed and tested. Modified to produce arbitrary beam patterns by controlling relief pattern. Antenna planar or contoured to supporting structure. Low off-axis radar cross section at frequencies removed from operational frequency. Interference pattern produced by spherical wave intersecting plane wave consists of concentric circles similar to Newton's rings. Pattern identical to Fresnel zone plate, which has lens properties. Plane wave incident on hologram, or zone plate, focused to point.

  18. Impact on multilayered composite plates

    NASA Technical Reports Server (NTRS)

    Kim, B. S.; Moon, F. C.

    1977-01-01

    Stress wave propagation in a multilayer composite plate due to impact was examined by means of the anisotropic elasticity theory. The plate was modelled as a number of identical anisotropic layers and the approximate plate theory of Mindlin was then applied to each layer to obtain a set of difference-differential equations of motion. Dispersion relations for harmonic waves and correction factors were found. The governing equations were reduced to difference equations via integral transforms. With given impact boundary conditions these equations were solved for an arbitrary number of layers in the plate and the transient propagation of waves was calculated by means of a Fast Fourier Transform algorithm. The multilayered plate problem was extended to examine the effect of damping layers present between two elastic layers. A reduction of the interlaminar normal stress was significant when the thickness of damping layer was increased but the effect was mostly due to the softness of the damping layer. Finally, the problem of a composite plate with a crack on the interlaminar boundary was formulated.

  19. Tracing the Farallon plate through seismic imaging with USArray

    NASA Astrophysics Data System (ADS)

    Porritt, Robert William

    The Farallon plate system has been subducting off the western United States since at least the middle Mesozoic. This plate has undergone virtually every subduction process during this time including a long episode of flat-slab subduction, generation of microplates, and formation of oceanic plateaus. The shallow remains of this plate are two small microplates, the Gorda and Juan de Fuca, in the Pacific Northwest. The anomalous nature of these two small plates and the missing deeper evidence of subduction has motivated this study. The USArray seismic experiment has provided unprecedented spatial sampling of the seismic wavefield in the continuous United States. Utilizing this dataset, new imaging methods have been implemented and older imaging methods have been revitalized. This study first uses ambient seismic noise in the Pacific Northwest to extract short period Rayleigh waves which are sensitive to lithospheric scale structure. Phase velocities from this model are then combined with teleseismic delay times of body waves and surface waves to image the structure of the continuous United States from the surface through the mantle transition zone. The resolving power of this model allows tracing of the Farallon plate from the trench to the lower mantle. The seismic velocity structure of the continuous United States is broadly composed of a slow western half and fast eastern half separated by the Rocky Mountain Front. The low velocity of the western U.S. contains several high velocity anomalies. While previous work has focused on individual anomalies and suggested they represent lithospheric instabilities, a larger regional view indicates that these are the western remnants of the Farallon plate. Below the thick cratonic lithosphere of the eastern U.S., the Farallon plate contains significant topography due to a subducted heterogeneity of the oceanic plate and a viscosity contrast through the mantle transition zone. The velocity models presented herein provide a

  20. Ultrasound source using a rectangular vibrating plate combined with rigid walls

    NASA Astrophysics Data System (ADS)

    Sato, Ryo; Asami, Takuya; Miura, Hikaru

    2017-07-01

    Ultrasound sources that use a stripe-mode rectangular vibrating plate radiate strong ultrasound waves in the air. In this study, we investigated the design strategy for combining the vibrating plate with rigid walls and evaluated the intense ultrasound waves radiated by the sound source. First, we examined the design method for a rectangular transverse vibrating plate with both ends fixed and the vibration amplitude distribution of the vibrating plate. Second, we measured the sound pressure distribution in the formation of the standing wave field. Finally, we clarified the relationship between the input power and sound pressure of the standing wave field antinodes.

  1. Model of waterlike fluid under confinement for hydrophobic and hydrophilic particle-plate interaction potentials.

    PubMed

    Krott, Leandro B; Barbosa, Marcia C

    2014-01-01

    Molecular dynamic simulations were employed to study a waterlike model confined between hydrophobic and hydrophilic plates. The phase behavior of this system is obtained for different distances between the plates and particle-plate potentials. For both hydrophobic and hydrophilic walls, there are the formation of layers. Crystallization occurs at lower temperature at the contact layer than at the middle layer. In addition, the melting temperature decreases as the plates become more hydrophobic. Similarly, the temperatures of maximum density and extremum diffusivity decrease with hydrophobicity.

  2. Pyrenean orogeny and plate kinematics

    NASA Astrophysics Data System (ADS)

    Sibuet, J.-C.; Srivastava, S.; Spakman, W.

    2003-04-01

    The development of the Pyrenees, a range of mountain which lies between Iberia and Eurasia, has remained a subject of much debate between geologists and geophysicists for a long time. The debate concerns about the large amount of compressive motion which geophysicists estimate took place across the Pyrenees based on the plate kinematics of the North Atlantic during the opening of the Bay of Biscay with which geologists differ based on their interpretations of the observations made across the Pyrenees. The geologists maintain that only a limited amount of Aptian-Albian compression can be supported by surface observations while a large amount of strike- slip motion is implied in their models. To find satisfactory answer to this disagreement we have re-examined not only the large amount of magnetic data which exist for the Bay of Biscay but also some of the deep seismic and teleseismic data across the Pyrenees. Our re-examination of magnetic data in the Bay of Biscay has enabled us to identify seafloor spreading anomalies 33o to M3. By combining these anomaly identifications with the existing identification of similar anomalies in the North Atlantic we have derived a position of a mean pole of rotation for the entire opening of the Bay of Biscay whose position is constrained not only by the magnetic anomalies identified in the Bay of Biscay and the North Atlantic but also by the geometry of a triple junction which had remained at the mouth of the Bay of Biscay during this period. Such a pole of rotation implies that about 400 km of shortening took place between the Iberian and Eurasian plates in the Pyrenean domain during the opening of the Bay of Biscay from chrons M0 to 33o time (118 to 80 Ma). Shortening perhaps took place in the form of subduction of the oceanic crust below Eurasia as reflected in the deep seismic ECORS profile shot across the Pyrenees. Careful examination of these data has shown presence of two deep features which dips to the north. Further

  3. How mantle slabs drive plate tectonics.

    PubMed

    Conrad, Clinton P; Lithgow-Bertelloni, Carolina

    2002-10-04

    The gravitational pull of subducted slabs is thought to drive the motions of Earth's tectonic plates, but the coupling between slabs and plates is not well established. If a slab is mechanically attached to a subducting plate, it can exert a direct pull on the plate. Alternatively, a detached slab may drive a plate by exciting flow in the mantle that exerts a shear traction on the base of the plate. From the geologic history of subduction, we estimated the relative importance of "pull" versus "suction" for the present-day plates. Observed plate motions are best predicted if slabs in the upper mantle are attached to plates and generate slab pull forces that account for about half of the total driving force on plates. Slabs in the lower mantle are supported by viscous mantle forces and drive plates through slab suction.

  4. Pacific plate motion change caused the Hawaiian-Emperor Bend.

    PubMed

    Torsvik, Trond H; Doubrovine, Pavel V; Steinberger, Bernhard; Gaina, Carmen; Spakman, Wim; Domeier, Mathew

    2017-06-05

    A conspicuous 60° bend of the Hawaiian-Emperor Chain in the north-western Pacific Ocean has variously been interpreted as the result of an abrupt Pacific plate motion change in the Eocene (∼47 Ma), a rapid southward drift of the Hawaiian hotspot before the formation of the bend, or a combination of these two causes. Palaeomagnetic data from the Emperor Seamounts prove ambiguous for constraining the Hawaiian hotspot drift, but mantle flow modelling suggests that the hotspot drifted 4-9° south between 80 and 47 Ma. Here we demonstrate that southward hotspot drift cannot be a sole or dominant mechanism for formation of the Hawaiian-Emperor Bend (HEB). While southward hotspot drift has resulted in more northerly positions of the Emperor Seamounts as they are observed today, formation of the HEB cannot be explained without invoking a prominent change in the direction of Pacific plate motion around 47 Ma.

  5. Pacific plate motion change caused the Hawaiian-Emperor Bend

    NASA Astrophysics Data System (ADS)

    Torsvik, Trond H.; Doubrovine, Pavel V.; Steinberger, Bernhard; Gaina, Carmen; Spakman, Wim; Domeier, Mathew

    2017-06-01

    A conspicuous 60° bend of the Hawaiian-Emperor Chain in the north-western Pacific Ocean has variously been interpreted as the result of an abrupt Pacific plate motion change in the Eocene (~47 Ma), a rapid southward drift of the Hawaiian hotspot before the formation of the bend, or a combination of these two causes. Palaeomagnetic data from the Emperor Seamounts prove ambiguous for constraining the Hawaiian hotspot drift, but mantle flow modelling suggests that the hotspot drifted 4-9° south between 80 and 47 Ma. Here we demonstrate that southward hotspot drift cannot be a sole or dominant mechanism for formation of the Hawaiian-Emperor Bend (HEB). While southward hotspot drift has resulted in more northerly positions of the Emperor Seamounts as they are observed today, formation of the HEB cannot be explained without invoking a prominent change in the direction of Pacific plate motion around 47 Ma.

  6. Characteristics and applications of advanced technology microchannel plates

    NASA Astrophysics Data System (ADS)

    Horton, Jerry R.; Tasker, G. William; Fijol, John J.

    1990-10-01

    A method for fabrication of novel thin-filrn continuous dynode electron multipliers is described. We have shown the feasibility of crucial manufacturing steps, including anisotropic dry etching of substrates into photolithographically-defined arrays of high-aspect-ratio channels, and the formation of thin-film continuous dynodes by chemical vapor deposition. We discuss potential performance and design advantages of this advanced technology microchannel plate (AT-MCP) over the conven tional reduced lead silicate glass inicrochannel plate (RLSG-'MCP) and implications for new applications.

  7. Phenomena after meteoroid penetration of a bumper plate

    NASA Technical Reports Server (NTRS)

    Todd, F. C.

    1971-01-01

    The analysis of hypervelocity impact of particles on a detector in space, with flow and shock penetration through fluid, plastic, and elastic zones was studied. The original paper and computer program on this topic is presented. Improvements in the program for the study of the formation of a cone of debris are discussed. The truncated apex of the cone is at the hole formed by the penetration of an initially spherical rock through a thin plate. A solution for the penetration of the thin plate was sought.

  8. Diluted honey inhibits biofilm formation: potential application in urinary catheter management?

    PubMed

    Emineke, Somadina; Cooper, Alan J; Fouch, Sarah; Birch, Brian R; Lwaleed, Bashir A

    2017-02-01

    Biofilms are ubiquitous and when mature have a complex structure of microcolonies in an extracellular polysaccharide and extracellular DNA matrix. Indwelling medical devices harbour biofilms which have been shown to cause infections and act as reservoirs for pathogens. Urinary catheters are often in place for considerable periods of time and are susceptible to both encrustation and biofilm formation. Strategies for minimising biofilm occurrence underpin an active research area in biomedicine. Manuka honey has, inter alia, well-established antibacterial properties. This study aims to assess the influence of honey on early biofilm formation in an established in vitro model. An established model of early biofilm formation using static bacterial cultures in vinyl 96-well plates was used to grow Escherichia coli, strain ATC 25922 and Proteus mirabilis, strain 7002. Planktonic cells were removed and the residual biofilm was stained with crystal violet, which were subsequently eluted and quantified spectrophotometrically. Manuka honey (Unique Manuka Factor 15+) was added either with the bacteria or up to 72 hours after. Biofilms in this model was developed over 3 days, after which growth stalled. Mixed (1:1) cultures of E. coli and P. mirabilis grew slower than monocultures. In mixed cultures, honey gave a dose-dependent reduction in biofilm formation (between 3.3 and 16.7%w/v). At 72 hours, all concentrations inhibited maximally (p<0.001). Application of honey to cultures after 24 and 48 hours also reduced the adherent bacterial biomass (p<0.05-p<0.01). Manuka honey at dilutions as low as 3.3% w/v in some protocols and at 10% or above in all protocols tested significantly inhibits bacterial attachment to a vinyl substrate and reduces further early biofilm development. No augmentation of growth over untreated controls was observed in any experiment. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence

  9. Geometry of the Cocos Plate Under North American Plate

    NASA Astrophysics Data System (ADS)

    Perez-Campos, X.

    2015-12-01

    The Cocos plate subducts under the North American plate with a complex geometry, and previous seismicity studies revealed some of this complexity. However, details of the geometry and the depth that the plate penetrates werelargely unknown. Since 2004, temporary experiments and the expansion of the permanent network of the Servicio Sismológico Nacional (SSN, Mexican National Seismological Service) have improved resolution of the plate geometry and have helped to map its descent into the upper mantle. Going from northwest to southeast, the Cocos plate appears to be fragmenting into north and south segments. The north segment subducts with an angle of ~30º and the south with an angle of ~10-15º. The transition is smooth near the trench and progresses to a tear at depth; this coincides with the projection of the Orozco Fracture Zone to depth. Also, this transition marks the limit of the presence to the south of an ultra slow velocity layer (USL) on top of the slab.South of this transition, the Cocos plate subducts horizontally , underplating the North American plate for a distance of ~140 to ~300 km from the trench. Along this horizontal region, silent slow events (SSE) and tectonic tremor (TT) have been observed. At a distance of 300 km from the trench (beneath central Mexico), the plate dives into the mantle with an angle of 76º to a depth of 500 km. This geometry changes abruptly to the south, marking the eastern limit of the USL. This change seems to be also characterized by a tear on the slab. Finally to the south, the Cocos plate subducts with a constant angle of 26º. This presentation summarizes the work of many contributors including A. Arciniega-Ceballos, M. Brudzinski, E. Cabral-Cano, T. Chen, R. Clayton,F. Cordoba-Montiel,P. Davis,S. Dougherty,F. Green, M. Gurnis, D. V. Helmberger, A. Husker,A. Iglesias, Y. Kim, V. Manea, D. Melgar, M. Rodríguez-Domínguez,S. K. Singh, T.-R. A. Song, C. M. Valdés-González, D. Valencia-Cabrera

  10. Plate mode velocities in graphite/epoxy plates

    NASA Technical Reports Server (NTRS)

    Prosser, W. H.; Gorman, M. R.

    1994-01-01

    Measurements of the velocities of the extensional and flexural plate modes were made along three directions of propagation in four graphite/epoxy composite plates. The acoustic signals were generated by simulated acoustic emission events (pencil lead breaks or Hsu-Neilson sources) and detected by by broadband ultrasonic transducers. The first arrival of the extensional plate mode, which is nondispersive at low frequencies, was measured at a number of different distances from the source along the propagation direction of interest. The velocity was determined by plotting the distance versus arrival time and computing its slope. Because of the large dispersion of the flexural mode, a Fourier phase velocity technique was used to characterize this mode. The velocity was measured up to a frequency of 160 kHz. Theoretical predictions of the velocities of these modes were also made and compared with experimental observations. Classical plate theory yields good agreement with the measured extensional velocities. For predictions of the dispersion of the flexural mode, Mindlin plates theory, which includes the effects of shear deformation and rotatory inertia was shown to give better agreement with the experimental measurements.

  11. Treatment of rabbit growth plate injuries with oriented ECM scaffold and autologous BMSCs

    PubMed Central

    Li, Wenchao; Xu, Ruijiang; Huang, Jiangxiang; Bao, Xing; Zhao, Bin

    2017-01-01

    Tissue-engineered technology has provided a promising method for the repair of growth plate injuries using biocompatible and biodegradable scaffolds and appropriate cells. The aim of this study was to fabricate oriented ECM scaffolds to imitate the material and structure of a natural growth plate and to investigate whether BMSCs in a scaffold could prevent the formation of bone bridges in an injured growth plate. We developed a natural, acellular and oriented scaffold derived from a growth plate. The oriented scaffold was fabricated using new freeze-drying technology and by cross-linking the microfilaments in the growth plate. From histological examination, the scaffold contained most of the ECM components including GAG and collagen II without cell DNA fragments, and SEM revealed that oriented scaffold had a uniform aperture in the transverse plane and columnar structure in length plane. Cytotoxicity testing with MTT showed no cytotoxic effect of the scaffold extracts on BMSCs. Autogenous BMSCs in oriented scaffolds promoted the regeneration of neogenetic growth plate when repairing an injured growth plate and prevent the formation of bone bridges to reduce the angular deformity and length discrepancy in the proximal tibia in rabbits. The well-characterized ECM-derived oriented growth plate scaffold shows potential for the repair of injured growth plates in young rabbits. PMID:28266598

  12. 3D high throughput screening and profiling of embryoid bodies in thermoformed microwell plates.

    PubMed

    Vrij, E J; Espinoza, S; Heilig, M; Kolew, A; Schneider, M; van Blitterswijk, C A; Truckenmüller, R K; Rivron, N C

    2016-02-21

    3D organoids using stem cells to study development and disease are now widespread. These models are powerful to mimic in vivo situations but are currently associated with high variability and low throughput. For biomedical research, platforms are thus necessary to increase reproducibility and allow high-throughput screens (HTS). Here, we introduce a microwell platform, integrated in standard culture plates, for functional HTS. Using micro-thermoforming, we form round-bottom microwell arrays from optically clear cyclic olefin polymer films, and assemble them with bottom-less 96-well plates. We show that embryonic stem cells aggregate faster and more reproducibly (centricity, circularity) as compared to a state-of-the-art microwell array. We then run a screen of a chemical library to direct differentiation into primitive endoderm (PrE) and, using on-chip high content imaging (HCI), we identify molecules, including regulators of the cAMP pathway, regulating tissue size, morphology and PrE gene activity. We propose that this platform will benefit to the systematic study of organogenesis in vitro.

  13. Earth's plate motion evolution and its link to global mantle dynamics