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Sample records for plate assay revealed

  1. An improved plating assay for determination of phage titer | Yang ...

    African Journals Online (AJOL)

    In this study, an improved plating assay was developed for detection of the number of recombinant phage Cap-T7 present in a test solution at a certain dilution point by counting the plaque forming units. The data demonstrated that the improved plating assay is fast, useful, and convenient for the determination of the phage ...

  2. Evaluation of immunoradiometric and ELISA versions of a microtitre plate assay for Bacillus anthracis spores

    Energy Technology Data Exchange (ETDEWEB)

    Phillips, A P; Martin, K L; Cross, N L [Chemical Defence Experimental Establishment, Porton (UK); Drake, R G [Glasgow Univ. (UK). Inst. of Biochemistry

    1984-05-11

    Solid-phase indirectly-labelled antibody assays for Bacillus anthracis spores heat-fixed on polystyrene microtitre plates were compared as immunoradiometric assay (IRMA) and enzyme-linked immunosorbent assay (ELISA) versions. Signal-to-noise ratios were usually higher in the IRMA than in the ELISA performed under parallel conditions but replicates were more varied in the IRMA. The antigen detection threshold and resolution limit calculated after regression analysis were broadly comparable in the 2 types of assay.

  3. Adaptation of the Nelson-Somogyi reducing-sugar assay to a microassay using microtiter plates.

    Science.gov (United States)

    Green, F; Clausen, C A; Highley, T L

    1989-11-01

    The Nelson-Somogyi assay for reducing sugars was adapted to microtiter plates. The primary advantages of this modified assay are (i) smaller sample and reagent volumes, (ii) elimination of boiling and filtration steps, (iii) automated measurement with a dual-wavelength scanning TLC densitometer, (iv) increased range and reproducibility, and (v) automated colorimetric readings by reflectance rather than absorbance.

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

    Science.gov (United States)

    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. An Assay in Microtitre Plates for Absolute Abundance of Chicken Interferon Alpha Transcripts

    Directory of Open Access Journals (Sweden)

    Renata Novak Kujundžić

    2010-01-01

    Full Text Available Immunosuppression of commercial chickens is a serious animal health and economic problem in the poultry industry. The major causes of the immunosuppression are viruses that suppress transcription of interferon genes, especially interferon alpha. There is a need for monitoring immunosuppression in commercially bred chickens. For this purpose, the absolute abundance of interferon alpha transcripts can be measured in blood of chickens by a suitable assay. Such an assay was used to estimate abundance of chicken interferon alpha in a sample of splenic cells induced with polyinosinic polycytidylic acid. The abundance measured was 29 ± 2 attomoles/µg total RNA. This assay can be performed in microtitre plates using samples collected from chickens in poultry houses.

  6. A novel assay reveals hygrotactic behavior in Drosophila.

    Directory of Open Access Journals (Sweden)

    Feiteng Ji

    Full Text Available Humidity is one of the most important factors that determines the geographical distribution and survival of terrestrial animals. The ability to detect variation in humidity is conserved across many species. Here, we established a novel behavioral assay that revealed the thirsty Drosophila exhibits strong hygrotactic behavior, and it can locate water by detecting humidity gradient. In addition, exposure to high levels of moisture was sufficient to elicit proboscis extension reflex behavior in thirsty flies. Furthermore, we found that the third antennal segment was necessary for hygrotactic behavior in thirsty flies, while arista was required for the avoidance of moist air in hydrated flies. These results indicated that two types of hygroreceptor cells exist in Drosophila: one located in the third antennal segment that mediates hygrotactic behavior in thirst status, and the other located in arista which is responsible for the aversive behavior toward moist air in hydration status. Using a neural silencing screen, we demonstrated that synaptic output from the mushroom body α/β surface and posterior neurons was required for both hygrotactic behavior and moisture-aversive behavior.

  7. Passive assay of plutonium metal plates using a fast-neutron multiplicity counter

    Energy Technology Data Exchange (ETDEWEB)

    Di Fulvio, A., E-mail: difulvio@umich.edu [Department of Nuclear Engineering & Radiological Sciences, University of Michigan, Ann Arbor, MI 48109 (United States); Shin, T.H.; Jordan, T.; Sosa, C.; Ruch, M.L.; Clarke, S.D. [Department of Nuclear Engineering & Radiological Sciences, University of Michigan, Ann Arbor, MI 48109 (United States); Chichester, D.L. [Idaho National Laboratory, Idaho Falls, ID 83415 (United States); Pozzi, S.A. [Department of Nuclear Engineering & Radiological Sciences, University of Michigan, Ann Arbor, MI 48109 (United States)

    2017-05-21

    We developed a fast-neutron multiplicity counter based on organic scintillators (EJ-309 liquid and stilbene). The system detects correlated photon and neutron multiplets emitted by fission reactions, within a gate time of tens of nanoseconds. The system was used at Idaho National Laboratory to assay a variety of plutonium metal plates. A coincidence counting strategy was used to quantify the {sup 240}Pu effective mass of the samples. Coincident neutrons, detected within a 40-ns coincidence window, show a monotonic trend, increasing with the {sup 240}Pu-effective mass (in this work, we tested the 0.005–0.5 kg range). After calibration, the system estimated the {sup 240}Pu effective mass of an unknown sample ({sup 240}Pu{sub eff} >50 g) with an uncertainty lower than 1% in a 4-min assay time.

  8. Micro-plate radiobinding assay of autoantibody to glutamic acid decarboxylase

    International Nuclear Information System (INIS)

    Huang Gan; Jin Helai; Wang Xia; Li Hui; Zhang Song; Zhou Zhiguang

    2008-01-01

    Objective: The purpose of this study was to develop a high-throughput micro-plate radiobinding assay (RBA) of glutamic acid decarboxylase antibody (CAD-Ab) and to evaluate its clinical application. Methods: 35 S labeled GAD 65 antigen was incubated with sera for 24 h on a 96-well plate, and then transferred to the Millipore plate coated with protein A, which was washed with 4 degree C PBS buffer, and then counted by a liquid scintillation counter. The CAD-Ab results were expressed by WHO standard unit (U/ml). A total of 224 healthy controls, 162 patients with type 1 diabetes mellitus (T1DM) and 210 patients with newly diagnosed type 2 diabetes (T2DM) were recruited. A total of 119 T1DM and healthy eases with gradually changing GAD-Ab levels were selected to compare the consistency of micro-plate RBA with conventional radioligand assay (RLA). Blood samples were obtained from the peripheral vein and finger tip in 32 healthy controls, 35 T1DM and 24 T2DM patients, and tested with micro-plate RBA and then compared with the conventional RLA to investigate the reliability of finger tip sampling. Linear correlation, student's t-test, variance analysis and receiver operating characteristic (ROC) curve were performed using SPSS 11.5. Results: (1) The optimized conditions of micro-plate RBA included 2 μl serum incubated with 3 x 10 4 counts/min 35 S-CAD for 24 h under slow vibration, antigen-antibody compounds washed 10 times by 4 degree C PBS buffer, and radioactivity counted with Optiphase Supermix scintillation liquid. (2)The intra-batch CV of the micro-plate RBA was 3.8%-10.2%, and the inter-batch CV was 5.6%-11.9%. The linearity analysis showed a good correlation when the GAD-Ab in serum samples ranged from 40.3 to 664 U/ml and the detection limit of measurement was 3.6 U/ml. The results from Diabetes Autoantibody Standardization Program (DASP) 2005 showed that the sensitivity and specificity for GAD-Ab were 78% (39 positive among 50 new-onset T1DM) and 98% (2 positive

  9. Understanding colonization and proliferation potential of endophytes and pathogen in planta via plating, polymerase chain reaction and ergosterol assay.

    Science.gov (United States)

    Chow, Yiing Yng; Rahman, Sadequr; Ting, Adeline Su Yien

    2017-01-01

    This study aimed to establish the colonization behavior and proliferation potential of three endophytes and one pathogen Ganoderma boninense (Gb) introduced into oil palm ramets (host model). The endophytes selected were Diaporthe phaseolorum (WAA02), Trichoderma asperellum (T2), and Penicillium citrinum (BTF08). Ramets were first inoculated with 100 mL of fungal cells (10 6  cfu mL - 1 ) via soil drenching. For the next 7 days, ramets were sampled and subjected to three different assays to detect and identify fungal colonization, and establish their proliferation potential in planta . Plate assay revealed the presence of endophytes in root, stem and leaf tissues within 7 days after inoculation. Polymerase Chain Reaction (PCR) detected and identified the isolates from the plant tissues. The ergosterol assay (via high performance liquid chromatography, HPLC) confirmed the presence of endophytes and Gb in planta . The increase in ergosterol levels throughout 49 days was however insignificant, suggesting that proliferation may be absent or may occur very slowly in planta . This study strongly suggests that the selected endophytes could colonize the host upon inoculation, but proliferation occurs at a slower rate, which may subsequently influence the biocontrol expression of endophytes against the pathogen.

  10. Understanding colonization and proliferation potential of endophytes and pathogen in planta via plating, polymerase chain reaction, and ergosterol assay

    Directory of Open Access Journals (Sweden)

    Yiing Yng Chow

    2017-01-01

    Full Text Available This study aimed to establish the colonization behavior and proliferation potential of three endophytes and one pathogen Ganoderma boninense (Gb introduced into oil palm ramets (host model. The endophytes selected were Diaporthe phaseolorum (WAA02, Trichoderma asperellum (T2, and Penicillium citrinum (BTF08. Ramets were first inoculated with 100 mL of fungal cells (106 cfu mL−1 via soil drenching. For the next 7 days, ramets were sampled and subjected to three different assays to detect and identify fungal colonization, and establish their proliferation potential in planta. Plate assay revealed the presence of endophytes in root, stem and leaf tissues within 7 days after inoculation. Polymerase Chain Reaction (PCR detected and identified the isolates from the plant tissues. The ergosterol assay (via high-performance liquid chromatography, HPLC confirmed the presence of endophytes and Gb in planta. The increase in ergosterol levels throughout 49 days was however insignificant, suggesting that proliferation may be absent or may occur very slowly in planta. This study strongly suggests that the selected endophytes could colonize the host upon inoculation, but proliferation occurs at a slower rate, which may subsequently influence the biocontrol expression of endophytes against the pathogen.

  11. Characterization of dermal plates from armored catfish Pterygoplichthys pardalis reveals sandwich-like nanocomposite structure.

    Science.gov (United States)

    Ebenstein, Donna; Calderon, Carlos; Troncoso, Omar P; Torres, Fernando G

    2015-05-01

    Dermal plates from armored catfish are bony structures that cover their body. In this paper we characterized structural, chemical, and nanomechanical properties of the dermal plates from the Amazonian fish Pterygoplichthys pardalis. Analysis of the morphology of the plates using scanning electron microscopy (SEM) revealed that the dermal plates have a sandwich-like structure composed of an inner porous matrix surrounded by two external dense layers. This is different from the plywood-like laminated structure of elasmoid fish scales but similar to the structure of osteoderms found in the dermal armour of some reptiles and mammals. Chemical analysis performed using Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) and X-ray diffraction (XRD) results revealed similarities between the composition of P. pardalis plates and the elasmoid fish scales of Arapaima gigas. Reduced moduli of P. pardalis plates measured using nanoindentation were also consistent with reported values for A. gigas scales, but further revealed that the dermal plate is an anisotropic and heterogeneous material, similar to many other fish scales and osteoderms. It is postulated that the sandwich-like structure of the dermal plates provides a lightweight and tough protective layer. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Homogeneous plate based antibody internalization assay using pH sensor fluorescent dye.

    Science.gov (United States)

    Nath, Nidhi; Godat, Becky; Zimprich, Chad; Dwight, Stephen J; Corona, Cesear; McDougall, Mark; Urh, Marjeta

    2016-04-01

    Receptor-mediated antibody internalization is a key mechanism underlying several anti-cancer antibody therapeutics. Delivering highly toxic drugs to cancer cells, as in the case of antibody drug conjugates (ADCs), efficient removal of surface receptors from cancer cells and changing the pharmacokinetics profile of the antibody drugs are some of key ways that internalization impacts the therapeutic efficacy of the antibodies. Over the years, several techniques have been used to study antibody internalization including radiolabels, fluorescent microscopy, flow cytometry and cellular toxicity assays. While these methods allow analysis of internalization, they have limitations including a multistep process and limited throughput and are generally endpoint assays. Here, we present a new homogeneous method that enables time and concentration dependent measurements of antibody internalization. The method uses a new hydrophilic and bright pH sensor dye (pHAb dye), which is not fluorescent at neutral pH but becomes highly fluorescent at acidic pH. For receptor mediated antibody internalization studies, antibodies against receptors are conjugated with the pHAb dye and incubated with the cells expressing the receptors. Upon binding to the receptor, the dyes conjugated to the antibody are not fluorescent because of the neutral pH of the media, but upon internalization and trafficking into endosomal and lysosomal vesicles the pH drops and dyes become fluorescent. The enabling attributes of the pHAb dyes are the hydrophilic nature to minimize antibody aggregation and bright fluorescence at acidic pH which allows development of simple plate based assays using a fluorescent reader. Using two different therapeutic antibodies--Trastuzumab (anti-HER2) and Cetuximab (anti-EGFR)--we show labeling with pHAb dye using amine and thiol chemistries and impact of chemistry and dye to antibody ration on internalization. We finally present two new approaches using the pHAb dye, which will be

  13. Standardization of the CFU-GM assay: Advantages of plating a fixed number of CD34+ cells in collagen gels.

    Science.gov (United States)

    Dobo, Irène; Pineau, Danielle; Robillard, Nelly; Geneviève, Frank; Piard, Nicole; Zandecki, Marc; Hermouet, Sylvie

    2003-10-01

    We investigated whether plating a stable amount of CD34(+) cells improves the CFU-GM assay. Data of CFU-GM assays performed with leukaphereses products in two transplant centers using a commercial collagen-based medium and unified CFU-GM scoring criteria were pooled and analyzed according to the numbers of CD34(+) cells plated. A first series of 113 CFU-GM assays was performed with a fixed number of mononuclear cells (i.e., a variable number of CD34(+) cells). In these cultures the CFU-GM/CD34 ratio varied according to the number of CD34(+) cells plated: median CFUGM/CD34 ratios were 1/6.2 to 1/6.6 for grafts containing or =2% CD34(+) cells. The median CFU-GM/CD34 ratio also varied depending on pathology: 1/9.3 for multiple myeloma (MM), 1/6.8 for Hodgkin's disease (HD), 1/6.5 for non-Hodgkin lymphoma (NHL), and 1/4.5 for solid tumors (ST). A second series of 95 CFU-GM assays was performed with a fixed number of CD34(+) cells (220/ml). The range of median CFU-GM/CD34 ratios was narrowed to 1/7.0 to 1/5.2, and coefficients of variation for CFU-GM counts decreased by half to 38.1% (NHL), 36.1% (MM), 49.9% (HD), and 22.4% (ST). In addition, CFU-GM scoring was facilitated as the percentages of cultures with >50 CFU/GM/ml decreased from 6.7% to 43.8% when a variable number of CD34(+) cells was plated, to 4.5% to 16.7% when 220 CD34(+) cells/ml were plated. Hence, plating a fixed number of CD34(+) cells in collagen gels improves the CFU-GM assay by eliminating cell number-related variability and reducing pathology-related variability in colony growth.

  14. Image-based ELISA on an activated polypropylene microtest plate--a spectrophotometer-free low cost assay technique.

    Science.gov (United States)

    Parween, Shahila; Nahar, Pradip

    2013-10-15

    In this communication, we report ELISA technique on an activated polypropylene microtest plate (APPµTP) as an illustrative example of a low cost diagnostic assay. Activated test zone in APPµTP binds a capture biomolecule through covalent linkage thereby, eliminating non-specific binding often prevalent in absorption based techniques. Efficacy of APPµTP is demonstrated by detecting human immunoglobulin G (IgG), human immunoglobulin E (IgE) and Aspergillus fumigatus antibody in patient's sera. Detection is done by taking the image of the assay solution by a desktop scanner and analyzing the color of the image. Human IgE quantification by color saturation in the image-based assay shows excellent correlation with absorbance-based assay (Pearson correlation coefficient, r=0.992). Significance of the relationship is seen from its p value which is 4.087e-11. Performance of APPµTP is also checked with respect to microtiter plate and paper-based ELISA. APPµTP can quantify an analyte as precisely as in microtiter plate with insignificant non-specific binding, a necessary prerequisite for ELISA assay. In contrast, paper-ELISA shows high non-specific binding in control sera (false positive). Finally, we have carried out ELISA steps on APPµTP by ultrasound waves on a sonicator bath and the results show that even in 8 min, it can convincingly differentiate a test sample from a control sample. In short, spectrophotometer-free image-based miniaturized ELISA on APPµTP is precise, reliable, rapid, and sensitive and could be a good substitute for conventional immunoassay procedures widely used in clinical and research laboratories. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Development of a high-throughput assay for measuring lipase activity using natural triacylglycerols coated on microtiter plates.

    Science.gov (United States)

    Serveau-Avesque, Carole; Verger, Robert; Rodriguez, Jorge A; Abousalham, Abdelkarim

    2013-09-21

    We have designed a convenient, specific, sensitive and continuous lipase assay based on the use of natural triacylglycerols (TAGs) from the Aleurites fordii seed oil which contains α-eleostearic acid (9,11,13,cis,trans,trans-octadecatrienoic acid) and which was coated in the wells of microtiter plates. The coated TAG film cannot be desorbed by the various buffers used during the lipase assay. Upon lipase action, α-eleostearic acid is liberated and desorbed from the interface and then solubilized into the micellar phase. Consequently, the UV absorbance of the α-eleostearic acid is considerably enhanced due to the transformation from an adsorbed to a water soluble state. The lipase activity can be measured continuously by recording the variations with time of the UV absorption spectra. The rate of lipolysis was monitored by measuring the increase of OD at 272 nm, which was found to be linear with time and directly proportional to the amount of added lipase. This microtiter plate lipase assay, based on coated TAGs, presents various advantages as compared to the classical systems: (i) coated TAGs on the microtiter plates could be stored for a long-time at 4 °C, (ii) higher sensitivity in lipase detection, (iii) good reproducibility, and (iv) increase of signal to noise ratio due to high UV absorption after transfer of α-eleostearic acid from an adsorbed to a soluble state. Low concentrations, down to 1 pg mL(-1) of pure Thermomyces lanuginosus or human pancreatic lipase, could be detected under standard assay conditions. The detection sensitivity of this coated method is around 1000 times higher as compared to those obtained with the classical emulsified systems. This continuous high throughput lipase assay could be used to screen new lipases and/or lipase inhibitors present in various biological samples.

  16. Development and Validation of a Microtiter Plate-Based Assay for Determination of Bacteriophage Host Range and Virulence

    Directory of Open Access Journals (Sweden)

    Yicheng Xie

    2018-04-01

    Full Text Available Bacteriophages, which are the natural predators of bacteria, have re-emerged as an attractive alternative to combat antibiotic resistant bacteria. Phages are highly specific at the species and strain level and measurement of the phage host range plays an important role in utilizing the phage as antimicrobials. The most common method for phage host range determination has been to spot phage lysates on soft agar overlays and observe plaque formation. In this study, a liquid culture-based assay was developed in a 96-well microtiter plate format to measure the phage host range and virulence for a collection of 15 Salmonella phages against a panel of 20 Salmonella strains representing 11 serovars. This method was compared to a traditional spot method. The majority of the host range results from two methods were in agreement including in cases where a bacterial strain was insensitive to the phage. Each method produced a false-negative result in 19/300 (6% of the measured phage-host combinations when compared to the other method. The spot method tended to indicate greater phage sensitivity than the microtiter assay even though direct comparisons of the response magnitude between the two methods is difficult since they operate on different mechanisms. The microtiter plate assay was able to provide data on both the phage host range and virulence in greater resolution in a high-throughput format.

  17. Development and Validation of a Microtiter Plate-Based Assay for Determination of Bacteriophage Host Range and Virulence.

    Science.gov (United States)

    Xie, Yicheng; Wahab, Laith; Gill, Jason J

    2018-04-12

    Bacteriophages, which are the natural predators of bacteria, have re-emerged as an attractive alternative to combat antibiotic resistant bacteria. Phages are highly specific at the species and strain level and measurement of the phage host range plays an important role in utilizing the phage as antimicrobials. The most common method for phage host range determination has been to spot phage lysates on soft agar overlays and observe plaque formation. In this study, a liquid culture-based assay was developed in a 96-well microtiter plate format to measure the phage host range and virulence for a collection of 15 Salmonella phages against a panel of 20 Salmonella strains representing 11 serovars. This method was compared to a traditional spot method. The majority of the host range results from two methods were in agreement including in cases where a bacterial strain was insensitive to the phage. Each method produced a false-negative result in 19/300 (6%) of the measured phage-host combinations when compared to the other method. The spot method tended to indicate greater phage sensitivity than the microtiter assay even though direct comparisons of the response magnitude between the two methods is difficult since they operate on different mechanisms. The microtiter plate assay was able to provide data on both the phage host range and virulence in greater resolution in a high-throughput format.

  18. Microtiter plate based colorimetric assay for characterization of dehalogenation activity of GAC/Fe0 composite

    DEFF Research Database (Denmark)

    Hwang, Yuhoon; Salatas, Apostolos; Mines, Paul D.

    2015-01-01

    of nZVI and its composite with granular activated carbon (GAC). The assay focused on analysis of reaction products rather than its mother compounds, which gives more accurate quantification of reductive activity. The colorimetric assays were developed to quantify three reaction products, ammonia......Even though nanoscale zero valent iron (nZVI) has been intensively studied for the treatment of a plethora of pollutants through reductive reaction, a quantification of nZVI reactivity has not been standardized. Here, we developed series of colorimetric assays for determining reductive activity...

  19. A novel high-throughput assay for islet respiration reveals uncoupling of rodent and human islets.

    Directory of Open Access Journals (Sweden)

    Jakob D Wikstrom

    Full Text Available The pancreatic beta cell is unique in its response to nutrient by increased fuel oxidation. Recent studies have demonstrated that oxygen consumption rate (OCR may be a valuable predictor of islet quality and long term nutrient responsiveness. To date, high-throughput and user-friendly assays for islet respiration are lacking. The aim of this study was to develop such an assay and to examine bioenergetic efficiency of rodent and human islets.The XF24 respirometer platform was adapted to islets by the development of a 24-well plate specifically designed to confine islets. The islet plate generated data with low inter-well variability and enabled stable measurement of oxygen consumption for hours. The F1F0 ATP synthase blocker oligomycin was used to assess uncoupling while rotenone together with myxothiazol/antimycin was used to measure the level of non-mitochondrial respiration. The use of oligomycin in islets was validated by reversing its effect in the presence of the uncoupler FCCP. Respiratory leak averaged to 59% and 49% of basal OCR in islets from C57Bl6/J and FVB/N mice, respectively. In comparison, respiratory leak of INS-1 cells and C2C12 myotubes was measured to 38% and 23% respectively. Islets from a cohort of human donors showed a respiratory leak of 38%, significantly lower than mouse islets.The assay for islet respiration presented here provides a novel tool that can be used to study islet mitochondrial function in a relatively high-throughput manner. The data obtained in this study shows that rodent islets are less bioenergetically efficient than human islets as well as INS1 cells.

  20. A simple and convenient microtiter plate assay for the detection of bactericidal antibodies to Vibrio cholerae O1 and Vibrio cholerae O139.

    Science.gov (United States)

    Boutonnier, Alain; Dassy, Bruno; Duménil, Rémy; Guénolé, Alain; Ratsitorahina, Maherisoa; Migliani, René; Fournier, Jean-Michel

    2003-12-01

    It is believed that the correlate of protection for cholera can be determined by the serum vibriocidal assay. The currently available vibriocidal assays, based on the conventional agar plating technique, are labor intensive. We developed a simple and convenient microtiter plate assay for the detection of vibriocidal antibodies that is equally as efficient for Vibrio cholerae O1 and for V. cholerae O139. The addition of succinate and neotetrazolium made it possible to measure the growth of surviving bacterial target cells by monitoring a color change. We evaluated assay parameters (target strains, growth of target cells, complement source and concentration) that may affect the reproducibility of the method for V. cholerae O139. The results obtained with the microtiter plate assay were uniformly similar to those obtained with the conventional agar plating assay, when testing both the Inaba and Ogawa serotypes of V. cholerae O1. The microtiter plate assay was also convenient for measuring the activity of animal sera and mouse monoclonal antibodies.

  1. Implementation and development of an automated, ultra-high-capacity, acoustic, flexible dispensing platform for assay-ready plate delivery.

    Science.gov (United States)

    Griffith, Dylan; Northwood, Roger; Owen, Paul; Simkiss, Ellen; Brierley, Andrew; Cross, Kevin; Slaney, Andrew; Davis, Miranda; Bath, Colin

    2012-10-01

    Compound management faces the daily challenge of providing high-quality samples to drug discovery. The advent of new screening technologies has seen demand for liquid samples move toward nanoliter ranges, dispensed by contactless acoustic droplet ejection. Within AstraZeneca, a totally integrated assay-ready plate production platform has been created to fully exploit the advantages of this technology. This enables compound management to efficiently deliver large throughputs demanded by high-throughput screening while maintaining regular delivery of smaller numbers of compounds in varying plate formats for cellular or biochemical concentration-response curves in support of hit and lead optimization (structure-activity relationship screening). The automation solution, CODA, has the capability to deliver compounds on demand for single- and multiple-concentration ranges, in batch sizes ranging from 1 sample to 2 million samples, integrating seamlessly into local compound and test management systems. The software handles compound orders intelligently, grouping test requests together dependent on output plate type and serial dilution ranges so that source compound vessels are shared among numerous tests, ensuring conservation of sample, reduced labware and costs, and efficiency of work cell logistics. We describe the development of CODA to address the customer demand, challenges experienced, learning made, and subsequent enhancements.

  2. An analytical approach to reduce between-plate variation in multiplex assays that measure antibodies to Plasmodium falciparum antigens.

    Science.gov (United States)

    Fang, Rui; Wey, Andrew; Bobbili, Naveen K; Leke, Rose F G; Taylor, Diane Wallace; Chen, John J

    2017-07-17

    Antibodies play an important role in immunity to malaria. Recent studies show that antibodies to multiple antigens, as well as, the overall breadth of the response are associated with protection from malaria. Yet, the variability and reliability of antibody measurements against a combination of malarial antigens using multiplex assays have not been well characterized. A normalization procedure for reducing between-plate variation using replicates of pooled positive and negative controls was investigated. Sixty test samples (30 from malaria-positive and 30 malaria-negative individuals), together with five pooled positive-controls and two pooled negative-controls, were screened for antibody levels to 9 malarial antigens, including merozoite antigens (AMA1, EBA175, MSP1, MSP2, MSP3, MSP11, Pf41), sporozoite CSP, and pregnancy-associated VAR2CSA. The antibody levels were measured in triplicate on each of 3 plates, and the experiments were replicated on two different days by the same technician. The performance of the proposed normalization procedure was evaluated with the pooled controls for the test samples on both the linear and natural-log scales. Compared with data on the linear scale, the natural-log transformed data were less skewed and reduced the mean-variance relationship. The proposed normalization procedure using pooled controls on the natural-log scale significantly reduced between-plate variation. For malaria-related research that measure antibodies to multiple antigens with multiplex assays, the natural-log transformation is recommended for data analysis and use of the normalization procedure with multiple pooled controls can improve the precision of antibody measurements.

  3. High-throughput micro plate vanillin assay for determination of tannin in sorghum grain

    Science.gov (United States)

    Sorghum tannins are phenolic compounds that offer health promoting antioxidant properties. The conventional HCl-vanillin assay for determining tannin content is a time-consuming method for screening large sample sets as seen in association mapping panels or breeder nursery samples. The objective of ...

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

    DEFF Research Database (Denmark)

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

    2008-01-01

    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......, 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...

  5. Tuning a 96-Well Microtiter Plate Fluorescence-Based Assay to Identify AGE Inhibitors in Crude Plant Extracts

    Directory of Open Access Journals (Sweden)

    Luc Séro

    2013-11-01

    Full Text Available Advanced glycation end-products (AGEs are involved in the pathogenesis of numerous diseases. Among them, cellular accumulation of AGEs contributes to vascular complications in diabetes. Besides using drugs to lower blood sugar, a balanced diet and the intake of herbal products potentially limiting AGE formation could be considered beneficial for patients’ health. The current paper presents a simple and cheap high-throughput screening (HTS assay based on AGE fluorescence and suitable for plant extract screening. We have already implemented an HTS assay based on vesperlysines-like fluorescing AGEs quickly (24 h formed from BSA and ribose under physiological conditions. However, interference was noted when fluorescent compounds and/or complex mixtures were tested. To overcome these problems and apply this HTS assay to plant extracts, we developed a technique for systematic quantification of both vesperlysines (λexc 370 nm; λem 440 nm and pentosidine-like (λexc 335 nm; λem 385 nm AGEs. In a batch of medicinal and food plant extracts, hits were selected as soon as fluorescence decreased under a fixed threshold for at least one wavelength. Hits revealed during this study appeared to contain well-known and powerful anti-AGE substances, thus demonstrating the suitability of this assay for screening crude extracts (0.1 mg/mL. Finally, quercetin was found to be a more powerful reference compound than aminoguanidine in such assay.

  6. Protein-carbohydrate complex reveals circulating metastatic cells in a microfluidic assay

    KAUST Repository

    Simone, Giuseppina

    2013-02-11

    Advances in carbohydrate sequencing technologies reveal the tremendous complexity of the glycome and the role that glycomics might have to bring insight into the biological functions. Carbohydrate-protein interactions, in particular, are known to be crucial to most mammalian physiological processes as mediators of cell adhesion and metastasis, signal transducers, and organizers of protein interactions. An assay is developed here to mimic the multivalency of biological complexes that selectively and sensitively detect carbohydrate-protein interactions. The binding of β-galactosides and galectin-3 - a protein that is correlated to the progress of tumor and metastasis - is examined. The efficiency of the assay is related to the expression of the receptor while anchoring to the interaction\\'s strength. Comparative binding experiments reveal molecular binding preferences. This study establishes that the assay is robust to isolate metastatic cells from colon affected patients and paves the way to personalized medicine. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Protein-carbohydrate complex reveals circulating metastatic cells in a microfluidic assay

    KAUST Repository

    Simone, Giuseppina; Malara, Natalia Maria; Trunzo, Valentina; Perozziello, Gerardo; Neužil, Pavel; Francardi, Marco; Roveda, Laura; Renne, Maria; Prati, Ubaldo; Mollace, Vincenzo; Manz, Andreas; Di Fabrizio, Enzo M.

    2013-01-01

    Advances in carbohydrate sequencing technologies reveal the tremendous complexity of the glycome and the role that glycomics might have to bring insight into the biological functions. Carbohydrate-protein interactions, in particular, are known to be crucial to most mammalian physiological processes as mediators of cell adhesion and metastasis, signal transducers, and organizers of protein interactions. An assay is developed here to mimic the multivalency of biological complexes that selectively and sensitively detect carbohydrate-protein interactions. The binding of β-galactosides and galectin-3 - a protein that is correlated to the progress of tumor and metastasis - is examined. The efficiency of the assay is related to the expression of the receptor while anchoring to the interaction's strength. Comparative binding experiments reveal molecular binding preferences. This study establishes that the assay is robust to isolate metastatic cells from colon affected patients and paves the way to personalized medicine. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection

    Directory of Open Access Journals (Sweden)

    Candice Tosi Michelon

    2011-03-01

    Full Text Available Direct smear examination using Ziehl-Neelsen staining for pulmonary tuberculosis (PTB diagnosis is inexpensive and easy to use, but has the major limitation of low sensitivity. Rapid molecular methods are becoming more widely available in centralized laboratories, but they depend on timely reporting of results and strict quality assurance obtainable only from costly commercial kits available in high burden nations. This study describes a pre-commercial colorimetric method, Detect-TB, for detecting Mycobacterium tuberculosis DNA in which an oligonucleotide probe is fixed onto wells of microwell plates and hybridized with biotinylated polymerase chain reaction amplification products derived from clinical samples. The probe is capable of hybridising with the IS6110 insertion element and was used to specifically recognise the M. tuberculosis complex. When combined with an improved silica-based DNA extraction method, the sensitivity of the test was 50 colony-forming units of the M. tuberculosis reference strain H37Rv. The results that were in agreement with reference detection methods were observed in 95.2% (453/476 of samples included in the analysis. Sensitivity and specificity for 301 induced sputum samples and 175 spontaneous sputum samples were 85% and 98%, and 94% and 100%, respectively. This colorimetric method showed similar specificity to that described for commercially available kits and may provide an important contribution for PTB diagnosis.

  9. Design of a titering assay for lentiviral vectors utilizing direct extraction of DNA from transduced cells in microtiter plates

    Directory of Open Access Journals (Sweden)

    Michele E Murphy

    2016-01-01

    Full Text Available Using lentiviral vector products in clinical applications requires an accurate method for measuring transduction titer. For vectors lacking a marker gene, quantitative polymerase chain reaction is used to evaluate the number of vector DNA copies in transduced target cells, from which a transduction titer is calculated. Immune Design previously described an integration-deficient lentiviral vector pseudotyped with a modified Sindbis virus envelope for use in cancer immunotherapy (VP02, of the ZVex platform. Standard protocols for titering integration-competent lentiviral vectors employ commercial spin columns to purify vector DNA from transduced cells, but such columns are not optimized for isolation of extrachromosomal (nonintegrated DNA. Here, we describe a 96-well transduction titer assay in which DNA extraction is performed in situ in the transduction plate, yielding quantitative recovery of extrachromosomal DNA. Vector titers measured by this method were higher than when commercial spin columns were used for DNA isolation. Evaluation of the method's specificity, linear range, and precision demonstrate that it is suitable for use as a lot release assay to support clinical trials with VP02. Finally, the method is compatible with titering both integrating and nonintegrating lentiviral vectors, suggesting that it may be used to evaluate the transduction titer for any lentiviral vector.

  10. An in vitro fatty acylation assay reveals a mechanism for Wnt recognition by the acyltransferase Porcupine.

    Science.gov (United States)

    Asciolla, James J; Miele, Matthew M; Hendrickson, Ronald C; Resh, Marilyn D

    2017-08-18

    Wnt proteins are a family of secreted signaling proteins that play key roles in regulating cell proliferation in both embryonic and adult tissues. Production of active Wnt depends on attachment of palmitoleate, a monounsaturated fatty acid, to a conserved serine by the acyltransferase Porcupine (PORCN). Studies of PORCN activity relied on cell-based fatty acylation and signaling assays as no direct enzyme assay had yet been developed. Here, we present the first in vitro assay that accurately recapitulates PORCN-mediated fatty acylation of a Wnt substrate. The critical feature is the use of a double disulfide-bonded Wnt peptide that mimics the two-dimensional structure surrounding the Wnt acylation site. PORCN-mediated Wnt acylation was abolished when the Wnt peptide was treated with DTT, and did not occur with a linear (non-disulfide-bonded) peptide, or when the double disulfide-bonded Wnt peptide contained Ala substituted for the Ser acylation site. We exploited this in vitro Wnt acylation assay to provide direct evidence that the small molecule LGK974, which is in clinical trials for managing Wnt-driven tumors, is a bona fide PORCN inhibitor whose IC 50 for inhibition of Wnt fatty acylation in vitro closely matches that for inhibition of Wnt signaling. Side-by-side comparison of PORCN and Hedgehog acyltransferase (HHAT), two enzymes that attach 16-carbon fatty acids to secreted proteins, revealed that neither enzyme will accept the other's fatty acyl-CoA or peptide substrates. These findings illustrate the unique enzyme-substrate selectivity exhibited by members of the membrane-bound O -acyl transferase family. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  11. Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR)

    DEFF Research Database (Denmark)

    Skeldal, Sune; Kjaergaard, Maj M; Alwasel, Saleh

    2015-01-01

    the vps10p domain receptor sortilin and the neurotrophin receptor p75(NTR). However, proNGF-induced receptor complex formation has been difficult to directly assess other than by western blotting. We here describe a fluorescence resonance energy transfer (FRET) based fluorescence plate reader assay...

  12. A simple micro-extraction plate assay for automated LC-MS/MS analysis of human serum 25-hydroxyvitamin D levels.

    Science.gov (United States)

    Geib, Timon; Meier, Florian; Schorr, Pascal; Lammert, Frank; Stokes, Caroline S; Volmer, Dietrich A

    2015-01-01

    This short application note describes a simple and automated assay for determination of 25-hydroxyvitamin D (25(OH)D) levels in very small volumes of human serum. It utilizes commercial 96-well micro-extraction plates with commercial 25(OH)D isotope calibration and quality control kits. Separation was achieved using a pentafluorophenyl liquid chromatography column followed by multiple reaction monitoring-based quantification on an electrospray triple quadrupole mass spectrometer. Emphasis was placed on providing a simple assay that can be rapidly established in non-specialized laboratories within days, without the need for laborious and time consuming sample preparation steps, advanced calibration or data acquisition routines. The analytical figures of merit obtained from this assay compared well to established assays. To demonstrate the applicability, the assay was applied to analysis of serum samples from patients with chronic liver diseases and compared to results from a routine clinical immunoassay. Copyright © 2015 John Wiley & Sons, Ltd.

  13. Revealing the radial characteristics of Q-plate generated vortex beams

    CSIR Research Space (South Africa)

    Sephton, Bereneice

    2016-07-01

    Full Text Available Q-plates (QP) have become ubiquitous in experiments requiring the generation of vortex beams since its development in 2006. It consequently follows that it is important to characterize the vortex beams created by this geometric-phase optical element...

  14. High throughput analysis of red wine and grape phenolics-adaptation and validation of methyl cellulose precipitable tannin assay and modified Somers color assay to a rapid 96 well plate format.

    Science.gov (United States)

    Mercurio, Meagan D; Dambergs, Robert G; Herderich, Markus J; Smith, Paul A

    2007-06-13

    The methyl cellulose precipitable (MCP) tannin assay and a modified version of the Somers and Evans color assay were adapted to high-throughput (HTP) analysis. To improve efficiency of the MCP tannin assay, a miniaturized 1 mL format and a HTP format using 96 well plates were developed. The Somers color assay was modified to allow the standardization of pH and ethanol concentrations of wine samples in a simple one-step dilution with a buffer solution, thus removing inconsistencies between wine matrices prior to analysis and allowing for its adaptation to a HTP format. Validation studies showed that all new formats were efficient, and results were reproducible and analogous to the original formats.

  15. Electrophoretic mobility shift assay reveals a novel recognition sequence for Setaria italica NAC protein.

    Science.gov (United States)

    Puranik, Swati; Kumar, Karunesh; Srivastava, Prem S; Prasad, Manoj

    2011-10-01

    The NAC (NAM/ATAF1,2/CUC2) proteins are among the largest family of plant transcription factors. Its members have been associated with diverse plant processes and intricately regulate the expression of several genes. Inspite of this immense progress, knowledge of their DNA-binding properties are still limited. In our recent publication,1 we reported isolation of a membrane-associated NAC domain protein from Setaria italica (SiNAC). Transactivation analysis revealed that it was a functionally active transcription factor as it could stimulate expression of reporter genes in vivo. Truncations of the transmembrane region of the protein lead to its nuclear localization. Here we describe expression and purification of SiNAC DNA-binding domain. We further report identification of a novel DNA-binding site, [C/G][A/T][T/A][G/C]TC[C/G][A/T][C/G][G/C] for SiNAC by electrophoretic mobility shift assay. The SiNAC-GST protein could bind to the NAC recognition sequence in vitro as well as to sequences where some bases had been reshuffled. The results presented here contribute to our understanding of the DNA-binding specificity of SiNAC protein.

  16. High-throughput screening of cellulase F mutants from multiplexed plasmid sets using an automated plate assay on a functional proteomic robotic workcell

    Directory of Open Access Journals (Sweden)

    Qureshi Nasib

    2006-05-01

    Full Text Available Abstract Background The field of plasmid-based functional proteomics requires the rapid assay of proteins expressed from plasmid libraries. Automation is essential since large sets of mutant open reading frames are being cloned for evaluation. To date no integrated automated platform is available to carry out the entire process including production of plasmid libraries, expression of cloned genes, and functional testing of expressed proteins. Results We used a functional proteomic assay in a multiplexed setting on an integrated plasmid-based robotic workcell for high-throughput screening of mutants of cellulase F, an endoglucanase from the anaerobic fungus Orpinomyces PC-2. This allowed us to identify plasmids containing optimized clones expressing mutants with improved activity at lower pH. A plasmid library of mutagenized clones of the celF gene with targeted variations in the last four codons was constructed by site-directed PCR mutagenesis and transformed into Escherichia coli. A robotic picker integrated into the workcell was used to inoculate medium in a 96-well deep well plate, combining the transformants into a multiplexed set in each well, and the plate was incubated on the workcell. Plasmids were prepared from the multiplexed culture on the liquid handler component of the workcell and used for in vitro transcription/translation. The multiplexed expressed recombinant proteins were screened for improved activity and stability in an azo-carboxymethylcellulose plate assay. The multiplexed wells containing mutants with improved activity were identified and linked back to the corresponding multiplexed cultures stored in glycerol. Spread plates were prepared from the glycerol stocks and the workcell was used to pick single colonies from the spread plates, prepare plasmid, produce recombinant protein, and assay for activity. The screening assay and subsequent deconvolution of the multiplexed wells resulted in identification of improved Cel

  17. Lithospheric flexure under the Hawaiian volcanic load: Internal stresses and a broken plate revealed by earthquakes

    Science.gov (United States)

    Klein, Fred W.

    2016-01-01

    Several lines of earthquake evidence indicate that the lithospheric plate is broken under the load of the island of Hawai`i, where the geometry of the lithosphere is circular with a central depression. The plate bends concave downward surrounding a stress-free hole, rather than bending concave upward as with past assumptions. Earthquake focal mechanisms show that the center of load stress and the weak hole is between the summits of Mauna Loa and Mauna Kea where the load is greatest. The earthquake gap at 21 km depth coincides with the predicted neutral plane of flexure where horizontal stress changes sign. Focal mechanism P axes below the neutral plane display a striking radial pattern pointing to the stress center. Earthquakes above the neutral plane in the north part of the island have opposite stress patterns; T axes tend to be radial. The M6.2 Honomu and M6.7 Kiholo main shocks (both at 39 km depth) are below the neutral plane and show radial compression, and the M6.0 Kiholo aftershock above the neutral plane has tangential compression. Earthquakes deeper than 20 km define a donut of seismicity around the stress center where flexural bending is a maximum. The hole is interpreted as the soft center where the lithospheric plate is broken. Kilauea's deep conduit is seismically active because it is in the ring of maximum bending. A simplified two-dimensional stress model for a bending slab with a load at one end yields stress orientations that agree with earthquake stress axes and radial P axes below the neutral plane. A previous inversion of deep Hawaiian focal mechanisms found a circular solution around the stress center that agrees with the model. For horizontal faults, the shear stress within the bending slab matches the slip in the deep Kilauea seismic zone and enhances outward slip of active flanks.

  18. Seismic potential of weak, near-surface faults revealed at plate tectonic slip rates.

    Science.gov (United States)

    Ikari, Matt J; Kopf, Achim J

    2017-11-01

    The near-surface areas of major faults commonly contain weak, phyllosilicate minerals, which, based on laboratory friction measurements, are assumed to creep stably. However, it is now known that shallow faults can experience tens of meters of earthquake slip and also host slow and transient slip events. Laboratory experiments are generally performed at least two orders of magnitude faster than plate tectonic speeds, which are the natural driving conditions for major faults; the absence of experimental data for natural driving rates represents a critical knowledge gap. We use laboratory friction experiments on natural fault zone samples at driving rates of centimeters per year to demonstrate that there is abundant evidence of unstable slip behavior that was not previously predicted. Specifically, weak clay-rich fault samples generate slow slip events (SSEs) and have frictional properties favorable for earthquake rupture. Our work explains growing field observations of shallow SSE and surface-breaking earthquake slip, and predicts that such phenomena should be more widely expected.

  19. Agarose gel shift assay reveals that calreticulin favors substrates with a quaternary structure in solution

    DEFF Research Database (Denmark)

    Boelt, Sanne Grundvad; Houen, Gunnar; Højrup, Peter

    2015-01-01

    Here we present an agarose gel shift assay that, in contrast to other electrophoresis approaches, is loaded in the center of the gel. This allows proteins to migrate in either direction according to their isoelectric points. Therefore, the presented assay enables a direct visualization, separation...... structure. It is also demonstrated that the agarose gel shift assay is useful in the study of other protein interactions and can be used as an alternative method to native polyacrylamide gel electrophoresis....... measure of interactions. Therefore, no interaction studies between calreticulin and substrates in solution have been investigated previously. The results presented here indicate that calreticulin has a preference for substrates with a quaternary structure and primarily β-sheets in their secondary...

  20. A Multiplexed Assay That Monitors Effects of Multiple Compound Treatment Times Reveals Candidate Immune-Enhancing Compounds.

    Science.gov (United States)

    Zhao, Ziyan; Henowitz, Liza; Zweifach, Adam

    2018-05-01

    We previously developed a flow cytometry assay that monitored lytic granule exocytosis in cytotoxic T lymphocytes stimulated by contacting beads coated with activating anti-CD3 antibodies. That assay was multiplexed in that responses of cells that did or did not receive the activating stimulus were distinguished via changes in light scatter accompanying binding of cells to beads, allowing us to discriminate compounds that activate responses on their own from compounds that enhance responses in cells that received the activating stimulus, all within a single sample. Here we add a second dimension of multiplexing by developing means to assess in a single sample the effects of treating cells with test compounds for different times. Bar-coding cells before adding them to test wells lets us determine compound treatment time while also monitoring activation status and response amplitude at the point of interrogation. This multiplexed assay is suitable for screening 96-well plates. We used it to screen compounds from the National Cancer Institute, identifying several compounds that enhance anti-LAMP1 responses. Multiple-treatment-time (MTT) screening enabled by bar-coding and read via high-throughput flow cytometry may be a generally useful method for facilitating the discovery of compounds of interest.

  1. A novel flow cytometric HTS assay reveals functional modulators of ATP binding cassette transporter ABCB6.

    Science.gov (United States)

    Polireddy, Kishore; Khan, Mohiuddin Md Taimur; Chavan, Hemantkumar; Young, Susan; Ma, Xiaochao; Waller, Anna; Garcia, Matthew; Perez, Dominique; Chavez, Stephanie; Strouse, Jacob J; Haynes, Mark K; Bologa, Cristian G; Oprea, Tudor I; Tegos, George P; Sklar, Larry A; Krishnamurthy, Partha

    2012-01-01

    ABCB6 is a member of the adenosine triphosphate (ATP)-binding cassette family of transporter proteins that is increasingly recognized as a relevant physiological and therapeutic target. Evaluation of modulators of ABCB6 activity would pave the way toward a more complete understanding of the significance of this transport process in tumor cell growth, proliferation and therapy-related drug resistance. In addition, this effort would improve our understanding of the function of ABCB6 in normal physiology with respect to heme biosynthesis, and cellular adaptation to metabolic demand and stress responses. To search for modulators of ABCB6, we developed a novel cell-based approach that, in combination with flow cytometric high-throughput screening (HTS), can be used to identify functional modulators of ABCB6. Accumulation of protoporphyrin, a fluorescent molecule, in wild-type ABCB6 expressing K562 cells, forms the basis of the HTS assay. Screening the Prestwick Chemical Library employing the HTS assay identified four compounds, benzethonium chloride, verteporfin, tomatine hydrochloride and piperlongumine, that reduced ABCB6 mediated cellular porphyrin levels. Validation of the identified compounds employing the hemin-agarose affinity chromatography and mitochondrial transport assays demonstrated that three out of the four compounds were capable of inhibiting ABCB6 mediated hemin transport into isolated mitochondria. However, only verteporfin and tomatine hydrochloride inhibited ABCB6's ability to compete with hemin as an ABCB6 substrate. This assay is therefore sensitive, robust, and suitable for automation in a high-throughput environment as demonstrated by our identification of selective functional modulators of ABCB6. Application of this assay to other libraries of synthetic compounds and natural products is expected to identify novel modulators of ABCB6 activity.

  2. A novel flow cytometric HTS assay reveals functional modulators of ATP binding cassette transporter ABCB6.

    Directory of Open Access Journals (Sweden)

    Kishore Polireddy

    Full Text Available ABCB6 is a member of the adenosine triphosphate (ATP-binding cassette family of transporter proteins that is increasingly recognized as a relevant physiological and therapeutic target. Evaluation of modulators of ABCB6 activity would pave the way toward a more complete understanding of the significance of this transport process in tumor cell growth, proliferation and therapy-related drug resistance. In addition, this effort would improve our understanding of the function of ABCB6 in normal physiology with respect to heme biosynthesis, and cellular adaptation to metabolic demand and stress responses. To search for modulators of ABCB6, we developed a novel cell-based approach that, in combination with flow cytometric high-throughput screening (HTS, can be used to identify functional modulators of ABCB6. Accumulation of protoporphyrin, a fluorescent molecule, in wild-type ABCB6 expressing K562 cells, forms the basis of the HTS assay. Screening the Prestwick Chemical Library employing the HTS assay identified four compounds, benzethonium chloride, verteporfin, tomatine hydrochloride and piperlongumine, that reduced ABCB6 mediated cellular porphyrin levels. Validation of the identified compounds employing the hemin-agarose affinity chromatography and mitochondrial transport assays demonstrated that three out of the four compounds were capable of inhibiting ABCB6 mediated hemin transport into isolated mitochondria. However, only verteporfin and tomatine hydrochloride inhibited ABCB6's ability to compete with hemin as an ABCB6 substrate. This assay is therefore sensitive, robust, and suitable for automation in a high-throughput environment as demonstrated by our identification of selective functional modulators of ABCB6. Application of this assay to other libraries of synthetic compounds and natural products is expected to identify novel modulators of ABCB6 activity.

  3. A Soluble Fluorescent Binding Assay Reveals PIP2 Antagonism of TREK-1 Channels

    Directory of Open Access Journals (Sweden)

    Cerrone Cabanos

    2017-08-01

    Full Text Available Lipid regulation of ion channels by low-abundance signaling lipids phosphatidylinositol 4,5-bisphosphate (PIP2 and phosphatidic acid (PA has emerged as a central cellular mechanism for controlling ion channels and the excitability of nerves. A lack of robust assays suitable for facile detection of a lipid bound to a channel has hampered the probing of the lipid binding sites and measuring the pharmacology of putative lipid agonists for ion channels. Here, we show a fluorescent PIP2 competition assay for detergent-purified potassium channels, including TWIK-1-related K+-channel (TREK-1. Anionic lipids PA and phosphatidylglycerol (PG bind dose dependently (9.1 and 96 μM, respectively and agonize the channel. Our assay shows PIP2 binds with high affinity (0.87 μM but surprisingly can directly antagonize TREK-1 in liposomes. We propose a model for TREK-1 lipid regulation where PIP2 can compete with PA and PG agonism based on the affinity of the lipid for a site within the channel.

  4. A spectrophotometer-based diffusivity assay reveals that diffusion hindrance of small molecules in extracellular matrix gels used in 3D cultures is dominated by viscous effects.

    Science.gov (United States)

    Galgoczy, Roland; Pastor, Isabel; Colom, Adai; Giménez, Alicia; Mas, Francesc; Alcaraz, Jordi

    2014-08-01

    The design of 3D culture studies remains challenging due to the limited understanding of extracellular matrix (ECM)-dependent hindered diffusion and the lack of simple diffusivity assays. To address these limitations, we set up a cost-effective diffusivity assay based on a Transwell plate and the spectrophotometer of a Microplate Reader, which are readily accessible to cell biology groups. The spectrophotometer-based assay was used to assess the apparent diffusivity D of FITC-dextrans with molecular weight (4-70kDa) spanning the physiological range of signaling factors in a panel of acellular ECM gels including Matrigel, fibrin and type I collagen. Despite their technical differences, D data exhibited ∼15% relative difference with respect to FRAP measurements. Our results revealed that diffusion hindrance of small particles is controlled by the enhanced viscosity of the ECM gel in conformance with the Stokes-Einstein equation rather than by geometrical factors. Moreover, we provided a strong rationale that the enhanced ECM viscosity is largely contributed to by unassembled ECM macromolecules. We also reported that gels with the lowest D exhibited diffusion hindrance closest to the large physiologic hindrance of brain tissue, which has a typical pore size much smaller than ECM gels. Conversely, sparse gels (≤1mg/ml), which are extensively used in 3D cultures, failed to reproduce the hindered diffusion of tissues, thereby supporting that dense (but not sparse) ECM gels are suitable tissue surrogates in terms of macromolecular transport. Finally, the consequences of reduced diffusivity in terms of optimizing the design of 3D culture experiments were addressed in detail. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. A rapid microtiter plate serum bactericidal assay method for determining serum complement-mediated killing of Mannheimia haemolytica.

    Science.gov (United States)

    Ayalew, Sahlu; Confer, Anthony W; Shrestha, Binu; Payton, Mark E

    2012-05-01

    In this study, we describe a rapid microtiter serum bactericidal assay (RMSBA) that can be used to measure the functionality of immune sera. It quantifies bactericidal activity of immune sera in the presence of complement against a homologous bacterium, M. haemolytica in this case. There is high correlation between data from RMSBA and standard complement-mediated bacterial killing assay (r=0.756; p<0.0001). The RMSBA activity of sera can be generated in less than 5 h instead of overnight incubation. RMSBA costs substantially less in terms of time, labor, and resources and is highly reproducible. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. High-throughput micro-plate HCL-vanillin assay for screening tannin content in sorghum grain

    Science.gov (United States)

    Sorghum contains tannin which is a phenolic compound that offers health promoting antioxidant capacity. The HCl-vanillin assay is a common and time consuming method for determining tannin content, but is not efficient for screening large sample sets as seen in association mapping panels or breeding ...

  7. A Direct Quantitative Agar-Plate Based Assay for Analysis of Pseudomonas protegens PF-5 Degradation of Polyurethane Films (Postprint)

    Science.gov (United States)

    2014-10-02

    was dissolved in methanol ; Irogran (1 g) was dissolved in 25 mL THF; and AS-P108 was prepared by adding 0.3 g of catalyst to 10 g of resin and then... Selected results are in Supple- mental Information. 3. Results 3.1. General growth characteristics of Pf-5 on M9/citrate agar plates Dilutions of P...minimal. An IR map with the color scale corresponding to the in- tensity of the 1735 cm1 carbonyl absorbance from the urethane and ester components is

  8. In planta assays involving epigenetically silenced genes reveal inhibition of cytosine methylation by genistein

    Directory of Open Access Journals (Sweden)

    Arase Sachiko

    2012-03-01

    Full Text Available Abstract Background Cytosine methylation is involved in epigenetic control of gene expression in a wide range of organisms. An increasing number of examples indicate that changing the frequency of cytosine methylation in the genome is a feasible tool to engineer novel traits in plants. Although demethylating effects of compounds have been analyzed in human cultured cells in terms of suppressing cancer, their effect in plant cells has not been analyzed extensively. Here, we developed in planta assay systems to detect inhibition of cytosine methylation using plants that contain a transgene transcriptionally silenced by an epigenetic mechanism. Results Seeds of two transgenic plants were used: a petunia line that has been identified as a revertant of the co-suppression of the chalcone synthase-A (CHS-A gene and contains CHS-A transgenes whose transcription is repressed; Nicotiana benthamiana plants that contain the green fluorescent protein (GFP reporter gene whose transcription is repressed through virus-induced transcriptional gene silencing. Seeds of these plants were sown on a medium that contained a demethylating agent, either 5-azacytidine or trichostatin A, and the restoration of the transcriptionally active state of the transgene was detected in seedlings. Using these systems, we found that genistein, a major isoflavonoid compound, inhibits cytosine methylation, thus restoring transgene transcription. Genistein also restored the transcription of an epigenetically silenced endogenous gene in Arabidopsis plants. Conclusions Our assay systems allowed us to assess the inhibition of cytosine methylation, in particular of maintenance of methylation, by compounds in plant cells. These results suggest a novel role of flavonoids in plant cells and that genistein is useful for modifying the epigenetic state of plant genomes.

  9. A combined disc method with resazurin agar plate assay for early phenotypic screening of KPC, MBL and OXA-48 carbapenemases among Enterobacteriaceae.

    Science.gov (United States)

    Teethaisong, Y; Eumkeb, G; Nakouti, I; Evans, K; Hobbs, G

    2016-08-01

    To validate a combined disc method along with resazurin chromogenic agar for early screening and differentiation of Klebsiella pneumoniae carbapenemase, metallo-β-lactamase and OXA-48 carbapenemase-producing Enterobacteriaceae. The combined disc test comprising of meropenem alone and with EDTA, phenylboronic acid or both EDTA and phenylboronic acid, and temocillin alone were evaluated with the resazurin chromogenic agar plate assay against a total of 86 molecularly confirmed Enterobacteriaceae clinical isolates (11 metallo-β-lactamases, eight Kl. pneumoniae carbapenemases, 11 OXA-48, 32 AmpC and 15 extended-spectrum-β-lactamase producers and nine co-producers of extended-spectrum-β-lactamase and AmpC). The inhibition zone diameters were measured and interpreted at 7 h for the presence of carbapenemase. All carbapenemase producers were phenotypically distinguished by this assay with 100% sensitivity and specificity. This early phenotypic method is very simple, inexpensive, and reliable in the detection and differentiation of carbapenemase-producing Enterobacteriaceae. It could be exploited in any microbiological laboratory for diagnosis of these recalcitrant bacteria. This assay poses excellent performance in discrimination of Kl. pneumoniae carbapenemase, metallo-β-lactamase and OXA-48 carbapenemases within 7 h, which is much faster than conventional disc diffusion methods. The rapid detection could help clinicians screen patients, control infection and provide epidemiological surveillance. © 2016 The Society for Applied Microbiology.

  10. Mechanism of SOS PR-domain autoinhibition revealed by single-molecule assays on native protein from lysate.

    Science.gov (United States)

    Lee, Young Kwang; Low-Nam, Shalini T; Chung, Jean K; Hansen, Scott D; Lam, Hiu Yue Monatrice; Alvarez, Steven; Groves, Jay T

    2017-04-28

    The guanine nucleotide exchange factor (GEF) Son of Sevenless (SOS) plays a critical role in signal transduction by activating Ras. Here we introduce a single-molecule assay in which individual SOS molecules are captured from raw cell lysate using Ras-functionalized supported membrane microarrays. This enables characterization of the full-length SOS protein, which has not previously been studied in reconstitution due to difficulties in purification. Our measurements on the full-length protein reveal a distinct role of the C-terminal proline-rich (PR) domain to obstruct the engagement of allosteric Ras independently of the well-known N-terminal domain autoinhibition. This inhibitory role of the PR domain limits Grb2-independent recruitment of SOS to the membrane through binding of Ras·GTP in the SOS allosteric binding site. More generally, this assay strategy enables characterization of the functional behaviour of GEFs with single-molecule precision but without the need for purification.

  11. Development of ultra-short PCR assay to reveal BRAF V600 mutation status in Thai colorectal cancer tissues.

    Science.gov (United States)

    Chat-Uthai, Nunthawut; Vejvisithsakul, Pichpisith; Udommethaporn, Sutthirat; Meesiri, Puttarakun; Danthanawanit, Chetiya; Wongchai, Yannawan; Teerapakpinyo, Chinachote; Shuangshoti, Shanop; Poungvarin, Naravat

    2018-01-01

    The protein kinase BRAF is one of the key players in regulating cellular responses to extracellular signals. Somatic mutations of the BRAF gene, causing constitutive activation of BRAF, have been found in various types of human cancers such as malignant melanoma, and colorectal cancer. BRAF V600E and V600K, most commonly observed mutations in these cancers, may predict response to targeted therapies. Many techniques suffer from a lack of diagnostic sensitivity in mutation analysis in clinical samples with a low cancer cell percentage or poor-quality fragmented DNA. Here we present allele-specific real-time PCR assay for amplifying 35- to 45-base target sequences in BRAF gene. Forward primer designed for BRAF V600E detection is capable of recognizing both types of BRAF V600E mutation, i.e. V600E1 (c.1799T>A) and V600E2 (c.1799_1800delTGinsAA), as well as complex tandem mutation caused by nucleotide changes in codons 600 and 601. We utilized this assay to analyze Thai formalin-fixed paraffin-embedded tissues. Forty-eight percent of 178 Thai colorectal cancer tissues has KRAS mutation detected by highly sensitive commercial assays. Although these DNA samples contain low overall yield of amplifiable DNA, our newly-developed assay successfully revealed BRAF V600 mutations in 6 of 93 formalin-fixed paraffin-embedded colorectal cancer tissues which KRAS mutation was not detected. Ultra-short PCR assay with forward mutation-specific primers is potentially useful to detect BRAF V600 mutations in highly fragmented DNA specimens from cancer patients.

  12. Genetic diversity and relationships in mulberry (genus Morus as revealed by RAPD and ISSR marker assays

    Directory of Open Access Journals (Sweden)

    Thangavelu K

    2004-01-01

    Full Text Available Abstract Background The genus Morus, known as mulberry, is a dioecious and cross-pollinating plant that is the sole food for the domesticated silkworm, Bombyx mori. Traditional methods using morphological traits for classification are largely unsuccessful in establishing the diversity and relationships among different mulberry species because of environmental influence on traits of interest. As a more robust alternative, PCR based marker assays including RAPD and ISSR were employed to study the genetic diversity and interrelationships among twelve domesticated and three wild mulberry species. Results RAPD analysis using 19 random primers generated 128 discrete markers ranging from 500–3000 bp in size. One-hundred-nineteen of these were polymorphic (92%, with an average of 6.26 markers per primer. Among these were a few putative species-specific amplification products which could be useful for germplasm classification and introgression studies. The ISSR analysis employed six anchored primers, 4 of which generated 93 polymorphic markers with an average of 23.25 markers per primer. Cluster analysis of RAPD and ISSR data using the WINBOOT package to calculate the Dice coefficient resulted into two clusters, one comprising polyploid wild species and the other with domesticated (mostly diploid species. Conclusion These results suggest that RAPD and ISSR markers are useful for mulberry genetic diversity analysis and germplasm characterization, and that putative species-specific markers may be obtained which can be converted to SCARs after further studies.

  13. Heritable differences in schooling behavior among threespine stickleback populations revealed by a novel assay.

    Directory of Open Access Journals (Sweden)

    Abigail R Wark

    2011-03-01

    Full Text Available Identifying the proximate and ultimate mechanisms of social behavior remains a major goal of behavioral biology. In particular, the complex social interactions mediating schooling behavior have long fascinated biologists, leading to theoretical and empirical investigations that have focused on schooling as a group-level phenomenon. However, methods to examine the behavior of individual fish within a school are needed in order to investigate the mechanisms that underlie both the performance and the evolution of schooling behavior. We have developed a technique to quantify the schooling behavior of an individual in standardized but easily manipulated social circumstances. Using our model school assay, we show that threespine sticklebacks (Gasterosteus aculeatus from alternative habitats differ in behavior when tested in identical social circumstances. Not only do marine sticklebacks show increased association with the model school relative to freshwater benthic sticklebacks, they also display a greater degree of parallel swimming with the models. Taken together, these data indicate that marine sticklebacks exhibit a stronger tendency to school than benthic sticklebacks. We demonstrate that these population-level differences in schooling tendency are heritable and are shared by individuals within a population even when they have experienced mixed-population housing conditions. Finally, we begin to explore the stimuli that elicit schooling behavior in these populations. Our data suggest that the difference in schooling tendency between marine and benthic sticklebacks is accompanied by differential preferences for social vs. non-social and moving vs. stationary shelter options. Our study thus provides novel insights into the evolution of schooling behavior, as well as a new experimental approach to investigate the genetic and neural mechanisms that underlie this complex social behavior.

  14. A novel whole-bacterial enzyme linked-immunosorbant assay to quantify Chlamydia trachomatis specific antibodies reveals distinct differences between systemic and genital compartments.

    Directory of Open Access Journals (Sweden)

    Hannah L Albritton

    Full Text Available Chlamydia trachomatis (CT is the leading sexually transmitted bacterial infection. The continued global burden of CT infection strongly predicates the need for a vaccine to supplement current chlamydial control programs. The correlates of protection against CT are currently unknown, but they must be carefully defined to guide vaccine design. The localized nature of chlamydial infection in columnar epithelial cells of the genital tract necessitates investigation of immunity at the site of infection. The purpose of this study was to develop a sensitive whole bacterial enzyme-linked immunosorbent assay (ELISA to quantify and compare CT-specific IgG and IgA in sera and genital secretions from CT-infected women. To achieve this, elementary bodies (EBs from two of the most common genital serovars (D and E were attached to poly-L-lysine-coated microtiter plates with glutaraldehyde. EB attachment and integrity were verified by the presence of outer membrane antigens and the absence of bacterial cytoplasmic antigens. EB-specific IgG and IgA standards were developed by pooling sera with high titers of CT-specific antibodies from infected women. Serum, endocervical and vaginal secretions, and endocervical cytobrush specimens from CT-infected women were used to quantify CT-specific IgG and IgA which were then normalized to total IgG and IgA, respectively. Analyses of paired serum and genital samples revealed significantly higher proportions of EB-specific antibodies in genital secretions compared to sera. Cervical and vaginal secretions and cytobrush specimens had similar proportions of EB-specific antibodies, suggesting any one of these genital sampling techniques could be used to quantify CT-specific antibodies when appropriate normalization methodologies are implemented. Overall, these results illustrate the need to investigate genital tract CT antibody responses, and our assay provides a useful quantitative tool to assess natural immunity in defined

  15. In vivo NAD assay reveals the intracellular NAD contents and redox state in healthy human brain and their age dependences

    Science.gov (United States)

    Zhu, Xiao-Hong; Lu, Ming; Lee, Byeong-Yeul; Ugurbil, Kamil; Chen, Wei

    2015-01-01

    NAD is an essential metabolite that exists in NAD+ or NADH form in all living cells. Despite its critical roles in regulating mitochondrial energy production through the NAD+/NADH redox state and modulating cellular signaling processes through the activity of the NAD+-dependent enzymes, the method for quantifying intracellular NAD contents and redox state is limited to a few in vitro or ex vivo assays, which are not suitable for studying a living brain or organ. Here, we present a magnetic resonance (MR) -based in vivo NAD assay that uses the high-field MR scanner and is capable of noninvasively assessing NAD+ and NADH contents and the NAD+/NADH redox state in intact human brain. The results of this study provide the first insight, to our knowledge, into the cellular NAD concentrations and redox state in the brains of healthy volunteers. Furthermore, an age-dependent increase of intracellular NADH and age-dependent reductions in NAD+, total NAD contents, and NAD+/NADH redox potential of the healthy human brain were revealed in this study. The overall findings not only provide direct evidence of declined mitochondrial functions and altered NAD homeostasis that accompany the normal aging process but also, elucidate the merits and potentials of this new NAD assay for noninvasively studying the intracellular NAD metabolism and redox state in normal and diseased human brain or other organs in situ. PMID:25730862

  16. An Ultra-High Fluorescence Enhancement and High Throughput Assay for Revealing Expression and Internalization of Chemokine Receptor CXCR4.

    Science.gov (United States)

    He, Hua; Wang, Xiaojuan; Cheng, Tiantian; Xia, Yongqing; Lao, Jun; Ge, Baosheng; Ren, Hao; Khan, Naseer Ullah; Huang, Fang

    2016-04-18

    Revealing chemokine receptor CXCR4 expression, distribution, and internalization levels in different cancers helps to evaluate cancer progression or prognosis and to set personalized treatment strategy. We here describe a sensitive and high-throughput immunoassay for determining CXCR4 expression and distribution in cancer cells. The assay is accessible to a wide range of users in an ordinary lab only by dip-coating poly(styrene-co-N-isopropylacrylamide) spheres on the glass substrate. The self- assembled spheres form three-dimensional photonic colloidal crystals which enhance the fluorescence of CF647 and Alexa Fluor 647 by a factor of up to 1000. CXCR4 in cells is detected by using the sandwich immunoassay, where the primary antibody recognizes CXCR4 and the secondary antibody is labeled with CF647. With the newly established assay, we quantified the total expression of CXCR4, its distribution on the cell membrane and cytoplasm, and revealed their internalization level upon SDF-1α activation in various cancer cells, even for those with extremely low expression level. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Synthesizing genome-wide association studies and expression microarray reveals novel genes that act in the human growth plate to modulate height.

    Science.gov (United States)

    Lui, Julian C; Nilsson, Ola; Chan, Yingleong; Palmer, Cameron D; Andrade, Anenisia C; Hirschhorn, Joel N; Baron, Jeffrey

    2012-12-01

    Previous meta-analysis of genome-wide association (GWA) studies has identified 180 loci that influence adult height. However, each GWA locus typically comprises a set of contiguous genes, only one of which presumably modulates height. We reasoned that many of the causative genes within these loci influence height because they are expressed in and function in the growth plate, a cartilaginous structure that causes bone elongation and thus determines stature. Therefore, we used expression microarray studies of mouse and rat growth plate, human disease databases and a mouse knockout phenotype database to identify genes within the GWAS loci that are likely required for normal growth plate function. Each of these approaches identified significantly more genes within the GWA height loci than at random genomic locations (P analysis strongly implicates 78 genes in growth plate function, including multiple genes that participate in PTHrP-IHH, BMP and CNP signaling, and many genes that have not previously been implicated in the growth plate. Thus, this analysis reveals a large number of novel genes that regulate human growth plate chondrogenesis and thereby contribute to the normal variations in human adult height. The analytic approach developed for this study may be applied to GWA studies for other common polygenic traits and diseases, thus providing a new general strategy to identify causative genes within GWA loci and to translate genetic associations into mechanistic biological insights.

  18. Functional assays and metagenomic analyses reveals differences between the microbial communities inhabiting the soil horizons of a Norway spruce plantation.

    Directory of Open Access Journals (Sweden)

    Stéphane Uroz

    Full Text Available In temperate ecosystems, acidic forest soils are among the most nutrient-poor terrestrial environments. In this context, the long-term differentiation of the forest soils into horizons may impact the assembly and the functions of the soil microbial communities. To gain a more comprehensive understanding of the ecology and functional potentials of these microbial communities, a suite of analyses including comparative metagenomics was applied on independent soil samples from a spruce plantation (Breuil-Chenue, France. The objectives were to assess whether the decreasing nutrient bioavailability and pH variations that naturally occurs between the organic and mineral horizons affects the soil microbial functional biodiversity. The 14 Gbp of pyrosequencing and Illumina sequences generated in this study revealed complex microbial communities dominated by bacteria. Detailed analyses showed that the organic soil horizon was significantly enriched in sequences related to Bacteria, Chordata, Arthropoda and Ascomycota. On the contrary the mineral horizon was significantly enriched in sequences related to Archaea. Our analyses also highlighted that the microbial communities inhabiting the two soil horizons differed significantly in their functional potentials according to functional assays and MG-RAST analyses, suggesting a functional specialisation of these microbial communities. Consistent with this specialisation, our shotgun metagenomic approach revealed a significant increase in the relative abundance of sequences related glycoside hydrolases in the organic horizon compared to the mineral horizon that was significantly enriched in glycoside transferases. This functional stratification according to the soil horizon was also confirmed by a significant correlation between the functional assays performed in this study and the functional metagenomic analyses. Together, our results suggest that the soil stratification and particularly the soil resource

  19. A comparison of titers of anti-Brucella antibodies of naturally infected and healthy vaccinated cattle by standard tube agglutination test, microtiter plate agglutination test, indirect hemagglutination assay, and indirect enzyme-linked immunosorbent assay

    Directory of Open Access Journals (Sweden)

    Anju Mohan

    2016-07-01

    Full Text Available Aim: We determined the antibody response in cattle naturally infected with brucellosis and normal healthy adult cattle vaccinated during calf hood with strain 19. Materials and Methods: The antibody titers were measured by standard tube agglutination test (STAT, microtiter plate agglutination test (MAT, indirect hemagglutination assay (IHA, and indirect enzyme-linked immunosorbent assay (iELISA as per standard protocols. Results: The mean STAT titers were 1.963±0.345 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was extremely significant (p<0.0001. The mean MAT titers were 2.244±0.727 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was very significant (p<0.005. The mean IHA titers in infected cattle were 2.284±0.574, and those in healthy vaccinated cattle were 1.200±0.155. The difference was extremely significant (p=0.0002. However, the difference in mean iELISA titers of infected cattle (1.3678±0.014 and healthy vaccinated cattle (1.367±0.014 was non-significant. The infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals. However, it cannot be ascertained whether these antibodies are due to vaccine or response to infection. Since the infected animals had been vaccinated earlier, the current infection may suggest that vaccination was unable to induce protective levels of antibody. The heightened antibody response after infection may also indicate a secondary immune response to the antigens common to the vaccine strain and wild Brucella organisms. Conclusion: The brucellosis infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals.

  20. Revealing the ability of a novel polysaccharide bioflocculant in bioremediation of heavy metals sensed in a Vibrio bioluminescence reporter assay.

    Science.gov (United States)

    Sajayan, Arya; Seghal Kiran, G; Priyadharshini, S; Poulose, Navya; Selvin, Joseph

    2017-09-01

    A bioflocculant-producing bacterial strain, designated MSI021, was isolated from the marine sponge Dendrilla nigra and demonstrated 94% flocculation activity in a kaolin clay suspension. MSI021 was identified as Bacillus cereus based on phylogenetic affiliation and biochemical characteristics. The purified extra-cellular bioflocculant was chemically elucidated as a polysaccharide molecule. The polysaccharide bioflocculant was stable under both acidic and alkaline conditions (pH 2.0-10.0) and temperatures up to 100 °C. The purified bioflocculant efficiently nucleated the formation of silver nanoparticles which showed broad spectrum antibacterial activity. The ability of the bioflocculant to remediate heavy metal toxicity was evaluated by measuring the inhibition of bioluminescence expression in Vibrio harveyi. Enrichment of heavy metals such as zinc, mercury and copper at concentrations of 1, 2 and 3 mM in culture media showed significant reduction of bioluminescence in Vibrio, whereas media enriched with heavy metals and bioflocculant showed dose dependent improvement in the expression of bioluminescence. The assay results demonstrated that the polysaccharide bioflocculant effectively mitigates heavy metal toxicity, thereby improving the expression of bioluminescence in Vibrio. This bioluminescence reporter assay can be developed into a high-throughput format to monitor and evaluate of heavy metal toxicity. The findings of this study revealed that a novel polysaccharide bioflocculant produced by a marine B. cereus demonstrated strong flocculating performance and was effective in nucleating the formation antibacterial silver nanoparticles and removing heavy metals. These results suggest that the MSI021 polysaccharide bioflocculant can be used to develop greener waste water treatment systems. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Paleoarchean bedrock lithologies across the Makhonjwa Mountains of South Africa and Swaziland linked to geochemical, magnetic and tectonic data reveal early plate tectonic genes flanking subduction margins

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    Maarten de Wit

    2018-05-01

    Full Text Available The Makhonjwa Mountains, traditionally referred to as the Barberton Greenstone Belt, retain an iconic Paleoarchean archive against which numerical models of early earth geodynamics can be tested. We present new geologic and structural maps, geochemical plots, geo- and thermo-chronology, and geophysical data from seven silicic, mafic to ultramafic complexes separated by major shear systems across the southern Makhonjwa Mountains. All reveal signs of modern oceanic back-arc crust and subduction-related processes. We compare the rates of processes determined from this data and balance these against plate tectonic and plume related models. Robust rates of both horizontal and vertical tectonic processes derived from the Makhonjwa Mountain complexes are similar, well within an order of magnitude, to those encountered across modern oceanic and orogenic terrains flanking Western Pacific-like subduction zones. We conclude that plate tectonics and linked plate-boundary processes were well established by 3.2–3.6 Ga. Our work provides new constraints for modellers with rates of a ‘basket’ of processes against which to test Paleoarchean geodynamic models over a time period close to the length of the Phanerozoic. Keywords: Paleoarchean, Barberton Greenstone Belt, Onverwacht Suite, Geologic bedrock and structural maps, Geochemistry and geophysics, Plate tectonics

  2. Effects of β-glucan polysaccharide revealed by the dominant lethal assay and micronucleus assays, and reproductive performance of male mice exposed to cyclophosphamide

    Directory of Open Access Journals (Sweden)

    Rodrigo Juliano Oliveira

    2014-01-01

    Full Text Available β-glucan is a well-known polysaccharide for its chemopreventive effect. This study aimed to evaluate the chemopreventive ability of β-glucan in somatic and germ cells through the dominant lethal and micronucleus assays, and its influence on the reproductive performance of male mice exposed to cyclophosphamide. The results indicate that β-glucan is capable of preventing changes in DNA in both germ cells and somatic ones. Changes in germ cells were evaluated by the dominant lethal assay and showed damage reduction percentages of 46.46% and 43.79% for the doses of 100 and 150 mg/kg. For the somatic changes, evaluated by micronucleus assay in peripheral blood cells in the first week of treatment, damage reduction percentages from 80.63-116.32% were found. In the fifth and sixth weeks, the percentage ranged from 10.20-52.54% and -0.95-62.35%, respectively. Besides the chemopreventive efficiency it appears that the β-glucan, when combined with cyclophosphamide, is able to improve the reproductive performance of males verified by the significant reduction in rates of post-implantation losses and reabsorption in the mating of nulliparous females with males treated with cyclophosphamide.

  3. Inhibition of thrombin by functionalized C60 nanoparticles revealed via in vitro assays and in silico studies.

    Science.gov (United States)

    Liu, Yanyan; Fu, Jianjie; Pan, Wenxiao; Xue, Qiao; Liu, Xian; Zhang, Aiqian

    2018-01-01

    The studies on the human toxicity of nanoparticles (NPs) are far behind the rapid development of engineered functionalized NPs. Fullerene has been widely used as drug carrier skeleton due to its reported low risk. However, different from other kinds of NPs, fullerene-based NPs (C 60 NPs) have been found to have an anticoagulation effect, although the potential target is still unknown. In the study, both experimental and computational methods were adopted to gain mechanistic insight into the modulation of thrombin activity by nine kinds of C 60 NPs with diverse surface chemistry properties. In vitro enzyme activity assays showed that all tested surface-modified C 60 NPs exhibited thrombin inhibition ability. Kinetic studies coupled with competitive testing using 3 known inhibitors indicated that six of the C 60 NPs, of greater hydrophobicity and hydrogen bond (HB) donor acidity or acceptor basicity, acted as competitive inhibitors of thrombin by directly interacting with the active site of thrombin. A simple quantitative nanostructure-activity relationship model relating the surface substituent properties to the inhibition potential was then established for the six competitive inhibitors. Molecular docking analysis revealed that the intermolecular HB interactions were important for the specific binding of C 60 NPs to the active site canyon, while the additional stability provided by the surface groups through van der Waals interaction also play a key role in the thrombin binding affinity of the NPs. Our results suggest that thrombin is a possible target of the surface-functionalized C 60 NPs relevant to their anticoagulation effect. Copyright © 2017. Published by Elsevier B.V.

  4. Structural variation of the oceanic Moho in the Pacific plate revealed by active-source seismic data

    Science.gov (United States)

    Ohira, Akane; Kodaira, Shuichi; Nakamura, Yasuyuki; Fujie, Gou; Arai, Ryuta; Miura, Seiichi

    2017-10-01

    The characteristics of the oceanic Moho are known to depend on various factors, such as seafloor spreading rate, crustal age, and accretionary processes at a ridge. However, the effect of local magmatic activities on the seismic signature of the Moho is poorly understood. Here an active-source reflection and refraction survey is used to investigate crustal structure and Moho characteristics along a >1000-km-long profile southeast of the Shatsky Rise in a Pacific Ocean basin formed from the Late Jurassic to Early Cretaceous and spanning the onset of Shatsky Rise volcanism. Although the seismic velocity structure estimated from the refraction data showed typical characteristics of the oceanic crust of the old Pacific plate, the appearance of the Moho reflections was spatially variable. We observed clear Moho reflections such as those to be expected where the spreading rate is fast to intermediate only at the southwestern end of the profile, whereas Moho reflections were diffuse, weak, or absent along other parts of the profile. The poor Moho reflections can be explained by the presence of a thick crust-mantle transition layer, which is temporally coincident with the formation of the Shatsky Rise. We inferred that the crust-mantle transition layer was formed by changes in on-axis accretion process or modification of the primary Moho by off-axis magmatism, induced by magmatic activity of the Shatsky Rise.

  5. Interdependence of initial cell density, drug concentration and exposure time revealed by real-time impedance spectroscopic cytotoxicity assay

    DEFF Research Database (Denmark)

    Caviglia, Claudia; Zor, Kinga; Canepa, Silvia

    2015-01-01

    We investigated the combined effect of the initial cell density (12 500, 35 000, 75 000, and 100 000 cells cm−2) and concentration of the anti-cancer drug doxorubicin on HeLa cells by performing timedependent cytotoxicity assays using real-time electrochemical impedance spectroscopy. A correlation...... between the rate of cell death and the initial cell seeding density was found at 2.5 μM doxorubicin concentration, whereas this was not observed at 5 or 100 μM. By sensing the changes in the cell–substrate interaction using impedance spectroscopy under static conditions, the onset of cytotoxicity...... was observed 5 h earlier than when using a standard colorimetric end-point assay (MTS) which measures changes in the mitochondrial metabolism. Furthermore, with the MTS assay no cytotoxicity was observed after 15 h of incubation with 2.5 μM doxorubicin, whereas the impedance showed at this time point cell...

  6. Development of a dual luciferase activity and fluorescamine protein assay adapted to a 384 micro-well plate format: Reducing variability in human luciferase transactivation cell lines aimed at endocrine active substances

    Science.gov (United States)

    Brennan, Jennifer; Tillitt, Donald E.

    2018-01-01

    There is a need to adapt cell bioassays to 384-well and 1536-well formats instead of the traditional 96-well format as high-throughput screening (HTS) demands increase. However, the sensitivity and performance of the bioassay must be re-verified in these higher micro-well plates, and verification of cell health must also be HT (high-throughput). We have adapted two commonly used human breast luciferase transactivation cell bioassays, the recently re-named estrogen agonist/antagonist screening VM7Luc4E2 cell bioassay (previously designated BG1Luc4E2) and the androgen/glucocorticoid screening MDA-kb2 cell bioassay, to 384-well formats for HTS of endocrine-active substances (EASs). This cost-saving adaptation includes a fast, accurate, and easy measurement of protein amount in each well via the fluorescamine assay with which to normalize luciferase activity of cell lysates without requiring any transfer of the cell lysates. Here we demonstrate that by accounting for protein amount in the cell lysates, antagonistic agents can easily be distinguished from cytotoxic agents in the MDA-kb2 and VM7Luc4E2 cell bioassays. Additionally, we demonstrate via the fluorescamine assay improved interpretation of luciferase activity in wells along the edge of the plate (the so-called “edge effect”), thereby increasing usable wells to the entire plate, not just interior wells.

  7. Selective activation of SHP2 activity by cisplatin revealed by a novel chemical probe-based assay

    International Nuclear Information System (INIS)

    Kuo, Chun-Chen; Chu, Chi-Yuan; Lin, Jing-Jer; Lo, Lee-Chiang

    2010-01-01

    Src homology-2 (SH2) domain-containing phosphatase 2 (SHP2) is known to participate in several different signaling pathways to mediate cell growth, survival, migration, and differentiation. However, due to the lack of proper analytical tools, it is unclear whether the phosphatase activity of SHP2 is activated in most studies. We have previously developed an activity-based probe LCL2 that formed covalent linkage with catalytically active protein tyrosine phosphatases (PTPs). Here, by combining LCL2 with a SHP2 specific antibody, we established an assay system that enables the direct monitoring of SHP2 activity upon cisplatin treatment of cancer cells. The protocol is advantageous over conventional colorimetric or in-gel PTP assays as it is specific and does not require the use of radioisotope reagents. Using this assay, we found SHP2 activity was selectively activated by cisplatin. Moreover, the activation of SHP2 appeared to be specific for cisplatin as other DNA damage agents failed to activate the activity. Although the role of SHP2 activation by cisplatin treatments is still unclear to us, our results provide the first direct evidence for the activation of SHP2 during cisplatin treatments. More importantly, the concept of using activity-based probe in conjunction with target-specific antibodies could be extended to other enzyme classes.

  8. Validation of FRET Assay for the Screening of Growth Inhibitors of Escherichia coli Reveals Elongasome Assembly Dynamics

    Science.gov (United States)

    van der Ploeg, René; Goudelis, Spyridon Theodoros; den Blaauwen, Tanneke

    2015-01-01

    The increase in antibiotic resistant bacteria demands the development of new antibiotics against preferably new targets. The common approach is to test compounds for their ability to kill bacteria or to design molecules that inhibit essential protein activities in vitro. In the first case, the mode of action of the drug is unknown and in the second case, it is not known whether the compound will pass the impermeable barrier of the bacterial envelope. We developed an assay that detects the target of a compound, as well as its ability to pass the membrane(s) simultaneously. The Escherichia coli cytoskeletal protein MreB recruits protein complexes (elongasomes) that are essential for cell envelope growth. An in cell Förster Resonance Energy Transfer (FRET) assay was developed to detect the interaction between MreB molecules and between MreB and the elongasome proteins RodZ, RodA and PBP2. Inhibition of the polymerization of MreB by S-(3,4-dichlorobenzyl) isothiourea (A22) or of the activity of PBP2 by mecilinam resulted in loss or reduction of all measured interactions. This suggests that the interactions between the elongasome proteins are governed by a combination of weak affinities and substrate availability. This validated in cell FRET assay can be used to screen for cell envelope growth inhibitors. PMID:26263980

  9. Validation of FRET Assay for the Screening of Growth Inhibitors of Escherichia coli Reveals Elongasome Assembly Dynamics

    Directory of Open Access Journals (Sweden)

    René van der Ploeg

    2015-07-01

    Full Text Available The increase in antibiotic resistant bacteria demands the development of new antibiotics against preferably new targets. The common approach is to test compounds for their ability to kill bacteria or to design molecules that inhibit essential protein activities in vitro. In the first case, the mode of action of the drug is unknown and in the second case, it is not known whether the compound will pass the impermeable barrier of the bacterial envelope. We developed an assay that detects the target of a compound, as well as its ability to pass the membrane(s simultaneously. The Escherichia coli cytoskeletal protein MreB recruits protein complexes (elongasomes that are essential for cell envelope growth. An in cell Förster Resonance Energy Transfer (FRET assay was developed to detect the interaction between MreB molecules and between MreB and the elongasome proteins RodZ, RodA and PBP2. Inhibition of the polymerization of MreB by S-(3,4-dichlorobenzyl isothiourea (A22 or of the activity of PBP2 by mecilinam resulted in loss or reduction of all measured interactions. This suggests that the interactions between the elongasome proteins are governed by a combination of weak affinities and substrate availability. This validated in cell FRET assay can be used to screen for cell envelope growth inhibitors.

  10. Enzymatic assays for detecting lactose and sucrose in urine to reveal intravenous drug abuse with emphasis on buprenorphine.

    Science.gov (United States)

    Keltanen, T; Mariottini, C; Walta, A M; Rahikainen, A L; Ojanperä, I

    2017-06-01

    Buprenorphine and methadone are commonly used medications for opioid maintenance treatment (OMT), using sublingual and oral administration, respectively. Although beneficial for OMT, these drugs can also be abused by intravenous administration. In intravenous abuse cases, the adjuvants lactose and sucrose are excreted in urine without hydrolysis to monosaccharides, since there are no disaccharidases in the blood. We validated enzymatic methods for the analysis of lactose and sucrose in urine. The analytical performance of both assays was considered appropriate for detecting intravenous drug abuse. The principle was proven by analyzing 93 postmortem (PM) urine samples for lactose, following comprehensive toxicological drug screening. In addition, 32 clinical urine samples from potential drug abusers were analyzed to assess the effect of PM changes on the assay. The mean level of lactose was low in clinical samples and relatively low in PM samples in which no drugs were found. Markedly elevated levels were seen in many of the buprenorphine positive samples, suggesting intravenous administration. Enzymatic methods could provide a simple and cost effective way to assess the intravenous administration of OMT drugs or drugs of abuse. Very high levels of glucose in urine may interfere with the assays. Furthermore, other causes for elevated urine disaccharides, such as hypolactasia and increased intestinal permeability, need to be considered in the interpretation of the results. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  11. Human HepaRG Cells can be Cultured in Hanging-drop Plates for Cytochrome P450 Induction and Function Assays.

    Science.gov (United States)

    Murayama, Norie; Usui, Takashi; Slawny, Nicky; Chesné, Christophe; Yamazaki, Hiroshi

    2015-01-01

    Recent guidance/guidelines for industry recommend that cytochrome P450 induction can be assessed using human hepatocyte enzyme activity and/or mRNA levels to evaluate potential drug- drug interactions. To evaluate time-dependent cytochrome P450 induction precisely, induction of CYP1A2, CYP2B6, and CYP3A4 mRNA was confirmed (>2-fold) by the treatment with omeprazole, phenobarbital, and rifampicin, respectively, for 24 or 48 h on day 3 from the start of culture. After 24 h, the fold induction of CYP1A2 with 3.6 and 1.8x10(4) HepaRG cells per well was lower than that for 7.2x10(4) cells. CYP1A2 induction levels at 24 h were higher than those after 48 h. In contrast, higher CYP2B6 inductions were confirmed after 48 h exposure than after 24 h, independent of the number of cells per well. To help reduce the use of human cryopreserved hepatocytes, typical P450-dependent enzyme activities were investigated in human HepaRG cells cultured in commercial hanging-drop plates. Newly designed 96-well hanging-drop plates were capable of maintaining human CYP3A-dependent midazolam hydroxylation activities for up to 4 days using only 10% of the recommended initial 7.2x10(4) cells per well. Favorable HepaRG function using hanging-drop plates was confirmed by detecting 1'- hydroxymidazolam O-glucuronide on day 3, suggesting an improvement over traditional control plates in which this metabolite can be detected for 24-well plates. These results suggest that the catalytic function and/or induction of CYP1A2, CYP2B6, and CYP3A4 can be readily assessed with reduced numbers of starting HepaRG cells cultured in three-dimensional cultures in drops prepared with hanging-drop plates.

  12. Paper microzone plates.

    Science.gov (United States)

    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. Chromatin immunoprecipitation assays revealed CREB and serine 133 phospho-CREB binding to the CART gene proximal promoter.

    Science.gov (United States)

    Rogge, George A; Shen, Li-Ling; Kuhar, Michael J

    2010-07-16

    Both over expression of cyclic AMP response element binding protein (CREB) in the nucleus accumbens (NAc), and intra-accumbal injection of cocaine- and amphetamine-regulated transcript (CART) peptides, have been shown to decrease cocaine reward. Also, over expression of CREB in the rat NAc increased CART mRNA and peptide levels, but it is not known if this was due to a direct action of P-CREB on the CART gene promoter. The goal of this study was to test if CREB and P-CREB bound directly to the CRE site in the CART promoter, using chromatin immunoprecipitation (ChIP) assays. ChIP assay with anti-CREB antibodies showed an enrichment of the CART promoter fragment containing the CRE region over IgG precipitated material, a non-specific control. Forskolin, which was known to increase CART mRNA levels in GH3 cells, was utilized to show that the drug increased levels of P-CREB protein and P-CREB binding to the CART promoter CRE-containing region. A region of the c-Fos promoter containing a CRE cis-regulatory element was previously shown to bind P-CREB, and it was used here as a positive control. These data suggest that the effects of CREB over expression on blunting cocaine reward could be, at least in part, attributed to the increased expression of the CART gene by direct interaction of P-CREB with the CART promoter CRE site, rather than by some indirect action. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  14. A quantitative multiplex nuclease protection assay reveals immunotoxicity gene expression profiles in the rabbit model for vaginal drug safety evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Fichorova, Raina N., E-mail: rfichorova@rics.bwh.harvard.edu [Laboratory of Genital Tract Biology, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women' s Hospital and Harvard Medical School, Boston, MA (United States); Mendonca, Kevin; Yamamoto, Hidemi S.; Murray, Ryan [Laboratory of Genital Tract Biology, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women' s Hospital and Harvard Medical School, Boston, MA (United States); Chandra, Neelima; Doncel, Gustavo F. [CONRAD, Department of Obstetrics and Gynecology, Eastern Virginia Medical School, Norfolk, VA (United States)

    2015-06-15

    Any vaginal product that alters the mucosal environment and impairs the immune barrier increases the risk of sexually transmitted infections, especially HIV infection, which thrives on mucosal damage and inflammation. The FDA-recommended rabbit vaginal irritation (RVI) model serves as a first line selection tool for vaginal products; however, for decades it has been limited to histopathology scoring, insufficient to select safe anti-HIV microbicides. In this study we incorporate to the RVI model a novel quantitative nuclease protection assay (qNPA) to quantify mRNA levels of 25 genes representing leukocyte differentiation markers, toll-like receptors (TLR), cytokines, chemokines, epithelial repair, microbicidal and vascular markers, by designing two multiplex arrays. Tissue sections were obtained from 36 rabbits (6 per treatment arm) after 14 daily applications of a placebo gel, saline, 4% nonoxynol-9 (N-9), and three combinations of the anti-HIV microbicides tenofovir (TFV) and UC781 in escalating concentrations (highest: 10% TFV + 2.5%UC781). Results showed that increased expression levels of toll-like receptor (TLR)-4, interleukin (IL)-1β, CXCL8, epithelial membrane protein (EMP)-1 (P < 0.05), and decreased levels of TLR2 (P < 0.05), TLR3 and bactericidal permeability increasing protein (BPI) (P < 0.001) were associated with cervicovaginal mucosal alteration (histopathology). Seven markers showed a significant linear trend predicting epithelial damage (up with CD4, IL-1β, CXCL8, CCL2, CCL21, EMP1 and down with BPI). Despite the low tissue damage RVI scores, the high-dose microbicide combination gel caused activation of HIV host cells (SLC and CD4) while N-9 caused proinflammatory gene upregulation (IL-8 and TLR4) suggesting a potential for increasing risk of HIV via different mechanisms depending on the chemical nature of the test product. - Highlights: • A transcriptome nuclease protection assay assessed microbicides for vaginal safety. • Biomarkers were

  15. A quantitative multiplex nuclease protection assay reveals immunotoxicity gene expression profiles in the rabbit model for vaginal drug safety evaluation

    International Nuclear Information System (INIS)

    Fichorova, Raina N.; Mendonca, Kevin; Yamamoto, Hidemi S.; Murray, Ryan; Chandra, Neelima; Doncel, Gustavo F.

    2015-01-01

    Any vaginal product that alters the mucosal environment and impairs the immune barrier increases the risk of sexually transmitted infections, especially HIV infection, which thrives on mucosal damage and inflammation. The FDA-recommended rabbit vaginal irritation (RVI) model serves as a first line selection tool for vaginal products; however, for decades it has been limited to histopathology scoring, insufficient to select safe anti-HIV microbicides. In this study we incorporate to the RVI model a novel quantitative nuclease protection assay (qNPA) to quantify mRNA levels of 25 genes representing leukocyte differentiation markers, toll-like receptors (TLR), cytokines, chemokines, epithelial repair, microbicidal and vascular markers, by designing two multiplex arrays. Tissue sections were obtained from 36 rabbits (6 per treatment arm) after 14 daily applications of a placebo gel, saline, 4% nonoxynol-9 (N-9), and three combinations of the anti-HIV microbicides tenofovir (TFV) and UC781 in escalating concentrations (highest: 10% TFV + 2.5%UC781). Results showed that increased expression levels of toll-like receptor (TLR)-4, interleukin (IL)-1β, CXCL8, epithelial membrane protein (EMP)-1 (P < 0.05), and decreased levels of TLR2 (P < 0.05), TLR3 and bactericidal permeability increasing protein (BPI) (P < 0.001) were associated with cervicovaginal mucosal alteration (histopathology). Seven markers showed a significant linear trend predicting epithelial damage (up with CD4, IL-1β, CXCL8, CCL2, CCL21, EMP1 and down with BPI). Despite the low tissue damage RVI scores, the high-dose microbicide combination gel caused activation of HIV host cells (SLC and CD4) while N-9 caused proinflammatory gene upregulation (IL-8 and TLR4) suggesting a potential for increasing risk of HIV via different mechanisms depending on the chemical nature of the test product. - Highlights: • A transcriptome nuclease protection assay assessed microbicides for vaginal safety. • Biomarkers were

  16. Revealing of endogenous Marinobufagin by an ultra-specific and sensitive UHPLC-MS/MS assay in pregnant women.

    Science.gov (United States)

    Lenaerts, Charline; Bond, Liz; Tuytten, Robin; Blankert, Bertrand

    2018-09-01

    Marinobufagenin (MBG) is a bufadienolide cardiac inotrope implicated in volume expansion-mediated hypertensive states including essential hypertension and preeclampsia (PE). Endogenous MBG is an inhibitor of the α1-isoform of Na + ,K + -ATPase with vasoconstrictive and cardiotonic properties, causing hypertension and natriuresis. Elevated endogenous MBG-like material levels have been described by immunoassays in salt-sensitive pregnant and preeclamptic rats as well as in preeclamptic human patients. The rise of endogenous MBG-like material appears prior the development of the main symptoms of PE, leading us to consider MBG as one of the potential biomarkers for PE. The weak specificity and the high variability of the published immunoassays gives no certification about endogenous MBG existence. This led us to set-up a highly specific and sensitive analytical method to detect MBG in plasma at low levels relying on liquid chromatography combined to mass spectrometry (UHPLC-MS/MS) with recording of 7 highly specific MRM transitions for MBG. Pure MBG standard used in the method development was obtained by purification from the Bufo marinus toad venom. d 3 -25-hydroxyvitamin D3 was used as internal standard. An increasing organic gradient with mobile phase A and B composed of 97:3 (v/v) H 2 O: MeOH and 50:45:5 (v/v/v) MeOH:IPA:H 2 O at pH 4.5 respectively was used on a Pursuit 3 PFP column (100 mm × 3 mm; 3 µm) to allow elution and separation of the plasmatic compounds. Chromatographic analyses of plasma samples were preceded by a precipitation of proteins pretreatment. The developed UHPLC-MS/MS assay has been applied to early-pregnant women plasma samples allowing us to investigate MBG plasma levels. Thanks to the high specificity of the assay we were able to authenticate and certify the presence of endogenous MBG in early-pregnant women plasma with the use of the 7 selected specific mass transitions. These pioneering preliminary results are giving a

  17. Heterogeneity of tumor chemosensitivity in ovarian epithelial cancer revealed using the adenosine triphosphate-tumor chemosensitivity assay.

    Science.gov (United States)

    Zhang, Jin; Li, Hongxia

    2015-05-01

    Ovarian cancer has a poor prognosis, primarily due to the heterogeneity in chemosensitivity among patients. In the present study, this heterogeneity was evaluated in ovarian epithelial cancer (OEC) using an in vitro adenosine triphosphate tumor chemosensitivity assay (ATP-TCA). Specimens were collected from 80 patients who underwent cytoreductive surgery. Viable ovarian cancer cells obtained from malignant tissues were tested for sensitivity to paclitaxel (PTX), carboplatin (CBP), topotecan (TPT), gemcitabine (GEM), docetaxel (TXT), etoposide, bleomycin and 4-hydroperoxycyclophosphamide using ATP-TCA. The sensitivity, specificity, positive predictive value and negative predictive value for the clinical chemotherapy sensitivity of OEC were 88.6, 77.8, 83 and 84.8%, respectively. PTX demonstrated the highest sensitivity of all agents tested (82.5% in all specimens, 85.7% in recurrent specimens), followed by CBP (58.8 and 60.7%, respectively). The sensitivities to PTX and docetaxel (PIII) or low-differentiated specimens, respectively. The present study indicated that ATP-TCA is an effective method for guiding the choice of chemotherapy drugs. Notable heterogeneity of chemosensitivity was observed in the OEC specimens.

  18. A chimera embryo assay reveals a decrease in embryonic cellular proliferation induced by sperm from X-irradiated male mice

    International Nuclear Information System (INIS)

    Obasaju, M.F.; Wiley, L.M.; Oudiz, D.J.; Raabe, O.; Overstreet, J.W.

    1989-01-01

    Male mice were divided into three experimental groups and a control group. Mice in the experimental groups received one of three doses of acute X irradiation (1.73, 0.29, and 0.05 Gy) and together with the control unirradiated mice were then mated weekly to unirradiated female mice for a 9-week experimental period. Embryos were recovered from the weekly matings at the four-cell stage and examined by the chimera assay for proliferative disadvantage. Aggregation chimeras were constructed of embryos from female mice mated to irradiated males (experimental embryos) and embryos from females mated to unexposed males (control embryos) and contained either one experimental embryo and one control embryo (heterologous chimera) or two control embryos (control chimera). The control embryo in heterologous chimeras and either embryo in control chimeras were prelabeled with the vital dye fluorescein isothiocyanate (FITC), and the chimeras were cultured for 40 h and viewed under phase-contrast and epifluorescence microscopy to obtain total embryo cell number and the cellular contribution from the FITC-labeled embryo. Experimental and control embryos that were cultured singly were also examined for embryo cell number at the end of the 40-h culture period. In control chimeras, the mean ratio of the unlabeled cells:total chimera cell number (henceforth referred to as ''mean ratio'') was 0.50 with little or no weekly variation over the 9-week experimental period. During Weeks 4-7, the mean ratios of heterologous chimeras differed significantly from the mean ratio of control chimeras with the greatest differences occurring during Week 7 (0.41 for chimeras of 0.05 Gy dose group, 0.40 for chimeras of the 0.29 Gy dose group, and 0.17 for chimeras of the 1.73 Gy dose group)

  19. A dual-color luciferase assay system reveals circadian resetting of cultured fibroblasts by co-cultured adrenal glands.

    Directory of Open Access Journals (Sweden)

    Takako Noguchi

    Full Text Available In mammals, circadian rhythms of various organs and tissues are synchronized by pacemaker neurons in the suprachiasmatic nucleus (SCN of the hypothalamus. Glucocorticoids released from the adrenal glands can synchronize circadian rhythms in other tissues. Many hormones show circadian rhythms in their plasma concentrations; however, whether organs outside the SCN can serve as master synchronizers to entrain circadian rhythms in target tissues is not well understood. To further delineate the function of the adrenal glands and the interactions of circadian rhythms in putative master synchronizing organs and their target tissues, here we report a simple co-culture system using a dual-color luciferase assay to monitor circadian rhythms separately in various explanted tissues and fibroblasts. In this system, circadian rhythms of organs and target cells were simultaneously tracked by the green-emitting beetle luciferase from Pyrearinus termitilluminans (ELuc and the red-emitting beetle luciferase from Phrixothrix hirtus (SLR, respectively. We obtained tissues from the adrenal glands, thyroid glands, and lungs of transgenic mice that expressed ELuc under control of the promoter from a canonical clock gene, mBmal1. The tissues were co-cultured with Rat-1 fibroblasts as representative target cells expressing SLR under control of the mBmal1 promoter. Amplitudes of the circadian rhythms of Rat-1 fibroblasts were potentiated when the fibroblasts were co-cultured with adrenal gland tissue, but not when co-cultured with thyroid gland or lung tissue. The phases of Rat-1 fibroblasts were reset by application of adrenal gland tissue, whereas the phases of adrenal gland tissue were not influenced by Rat-1 fibroblasts. Furthermore, the effect of the adrenal gland tissue on the fibroblasts was blocked by application of a glucocorticoid receptor (GR antagonist. These results demonstrate that glucocorticoids are strong circadian synchronizers for fibroblasts and that

  20. Novel cell-based assay reveals associations of circulating serum AhR-ligands with metabolic syndrome and mitochondrial dysfunction.

    Science.gov (United States)

    Park, Wook-Ha; Jun, Dae Won; Kim, Jin Taek; Jeong, Jae Hoon; Park, Hyokeun; Chang, Yoon-Seok; Park, Kyong Soo; Lee, Hong Kyu; Pak, Youngmi Kim

    2013-01-01

    Serum concentrations of environmental pollutants have been positively correlated with diabetes and metabolic syndrome in epidemiologic studies. In turn, abnormal mitochondrial function has been associated with the diseases. The relationships between these variables, however, have not been studied. We developed novel cell-based aryl hydrocarbon receptor (AhR) agonist bioassay system without solvent extraction process and analyzed whether low-dose circulating AhR ligands in human serum are associated with parameters of metabolic syndrome and mitochondrial function. Serum AhR ligand activities were measured as serum 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalent (sTCDDeq) in pM using 10 μL human sera from 97 Korean participants (47 with glucose intolerance and 50 matched controls, average age of 46.6 ± 9.9 years, 53 male and 45 female). sTCDDeq were higher in participants with glucose intolerance than normal controls and were positively associated (P fasting glucose, but not with HDL-cholesterol. Body mass index was in a positive linear relationship with serum AhR ligands in healthy participants. When myoblast cells were incubated with human sera, ATP generating power of mitochondria became impaired in an AhR ligand concentration-dependent manner. Our results support that circulating AhR ligands may directly reduce mitochondrial function in tissues, leading to weight gain, glucose intolerance, and metabolic syndrome. Our rapid cell-based assay using minute volume of human serum may provide one of the best monitoring systems for circulating AhR ligands, good clinical biomarkers for the progress of disease and therapeutic efficacy. Copyright © 2013 International Union of Biochemistry and Molecular Biology, Inc.

  1. Presynaptic selectivity of a ligand for serotonin 1A receptors revealed by in vivo PET assays of rat brain.

    Directory of Open Access Journals (Sweden)

    Takeaki Saijo

    Full Text Available A novel investigational antidepressant with high affinity for the serotonin transporter and the serotonin 1A (5-HT(1A receptor, called Wf-516 (structural formula: (2S-1-[4-(3,4-dichlorophenylpiperidin-1-yl]-3-[2-(5-methyl-1,3,4-oxadiazol-2-ylbenzo[b]furan-4-yloxy]propan-2-ol monohydrochloride, has been found to exert a rapid therapeutic effect, although the mechanistic basis for this potential advantage remains undetermined. We comparatively investigated the pharmacokinetics and pharmacodynamics of Wf-516 and pindolol by positron emission tomographic (PET and autoradiographic assays of rat brains in order to elucidate their molecular interactions with presynaptic and postsynaptic 5-HT(1A receptors. In contrast to the full receptor occupancy by pindolol in PET measurements, the binding of Wf-516 to 5-HT(1A receptors displayed limited capacity, with relatively high receptor occupancy being achieved in regions predominantly containing presynaptic receptors. This selectivity was further proven by PET scans of neurotoxicant-treated rats deficient in presynaptic 5-HT(1A receptors. In addition, [(35S]guanosine 5'-O-[γ-thio]triphosphate autoradiography indicated a partial agonistic ability of Wf-516 for 5-HT(1A receptors. This finding has lent support to reports that diverse partial agonists for 5-HT(1A receptors exert high sensitivity for presynaptic components. Thus, the present PET data suggest a relatively high capacity of presynaptic binding sites for partial agonists. Since our in vitro and ex vivo autoradiographies failed to illustrate these distinct features of Wf-516, in vivo PET imaging is considered to be, thus far, the sole method capable of pharmacokinetically demonstrating the unique actions of Wf-516 and similar new-generation antidepressants.

  2. Presynaptic selectivity of a ligand for serotonin 1A receptors revealed by in vivo PET assays of rat brain.

    Science.gov (United States)

    Saijo, Takeaki; Maeda, Jun; Okauchi, Takashi; Maeda, Jun-ichi; Morio, Yasunori; Kuwahara, Yasuhiro; Suzuki, Masayuki; Goto, Nobuharu; Fukumura, Toshimitsu; Suhara, Tetsuya; Higuchi, Makoto

    2012-01-01

    A novel investigational antidepressant with high affinity for the serotonin transporter and the serotonin 1A (5-HT(1A)) receptor, called Wf-516 (structural formula: (2S)-1-[4-(3,4-dichlorophenyl)piperidin-1-yl]-3-[2-(5-methyl-1,3,4-oxadiazol-2-yl)benzo[b]furan-4-yloxy]propan-2-ol monohydrochloride), has been found to exert a rapid therapeutic effect, although the mechanistic basis for this potential advantage remains undetermined. We comparatively investigated the pharmacokinetics and pharmacodynamics of Wf-516 and pindolol by positron emission tomographic (PET) and autoradiographic assays of rat brains in order to elucidate their molecular interactions with presynaptic and postsynaptic 5-HT(1A) receptors. In contrast to the full receptor occupancy by pindolol in PET measurements, the binding of Wf-516 to 5-HT(1A) receptors displayed limited capacity, with relatively high receptor occupancy being achieved in regions predominantly containing presynaptic receptors. This selectivity was further proven by PET scans of neurotoxicant-treated rats deficient in presynaptic 5-HT(1A) receptors. In addition, [(35)S]guanosine 5'-O-[γ-thio]triphosphate autoradiography indicated a partial agonistic ability of Wf-516 for 5-HT(1A) receptors. This finding has lent support to reports that diverse partial agonists for 5-HT(1A) receptors exert high sensitivity for presynaptic components. Thus, the present PET data suggest a relatively high capacity of presynaptic binding sites for partial agonists. Since our in vitro and ex vivo autoradiographies failed to illustrate these distinct features of Wf-516, in vivo PET imaging is considered to be, thus far, the sole method capable of pharmacokinetically demonstrating the unique actions of Wf-516 and similar new-generation antidepressants.

  3. Stem cell-like differentiation potentials of endometrial side population cells as revealed by a newly developed in vivo endometrial stem cell assay.

    Directory of Open Access Journals (Sweden)

    Kaoru Miyazaki

    Full Text Available Endometrial stem/progenitor cells contribute to the cyclical regeneration of human endometrium throughout a woman's reproductive life. Although the candidate cell populations have been extensively studied, no consensus exists regarding which endometrial population represents the stem/progenitor cell fraction in terms of in vivo stem cell activity. We have previously reported that human endometrial side population cells (ESP, but not endometrial main population cells (EMP, exhibit stem cell-like properties, including in vivo reconstitution of endometrium-like tissues when xenotransplanted into immunodeficient mice. The reconstitution efficiency, however, was low presumably because ESP cells alone could not provide a sufficient microenvironment (niche to support their stem cell activity. The objective of this study was to establish a novel in vivo endometrial stem cell assay employing cell tracking and tissue reconstitution systems and to examine the stem cell properties of ESP through use of this assay.ESP and EMP cells isolated from whole endometrial cells were infected with lentivirus to express tandem Tomato (TdTom, a red fluorescent protein. They were mixed with unlabeled whole endometrial cells and then transplanted under the kidney capsule of ovariectomized immunodeficient mice. These mice were treated with estradiol and progesterone for eight weeks and nephrectomized. All of the grafts reconstituted endometrium-like tissues under the kidney capsules. Immunofluorescence revealed that TdTom-positive cells were significantly more abundant in the glandular, stromal, and endothelial cells of the reconstituted endometrium in mice transplanted with TdTom-labeled ESP cells than those with TdTom-labeled EMP cells.We have established a novel in vivo endometrial stem cell assay in which multi-potential differentiation can be identified through cell tracking during in vivo endometrial tissue reconstitution. Using this assay, we demonstrated that ESP

  4. Nuclear protein accumulation in cellular senescence and organismal aging revealed with a novel single-cell resolution fluorescence microscopy assay.

    Science.gov (United States)

    De Cecco, Marco; Jeyapalan, Jessie; Zhao, Xiaoai; Tamamori-Adachi, Mimi; Sedivy, John M

    2011-10-01

    Replicative cellular senescence was discovered some 50 years ago. The phenotypes of senescent cells have been investigated extensively in cell culture, and found to affect essentially all aspects of cellular physiology. The relevance of cellular senescence in the context of age-associated pathologies as well as normal aging is a topic of active and ongoing interest. Considerable effort has been devoted to biomarker discovery to enable the microscopic detection of single senescent cells in tissues. One characteristic of senescent cells documented very early in cell culture studies was an increase in cell size and total protein content, but whether this occurs in vivo is not known. A limiting factor for studies of protein content and localization has been the lack of suitable fluorescence microscopy tools. We have developed an easy and flexible method, based on the merocyanine dye known as NanoOrange, to visualize and quantitatively measure total protein levels by high resolution fluorescence microscopy. NanoOrange staining can be combined with antibody-based immunofluorescence, thus providing both specific target and total protein information in the same specimen. These methods are optimally combined with automated image analysis platforms for high throughput analysis. We document here increasing protein content and density in nuclei of senescent human and mouse fibroblasts in vitro, and in liver nuclei of aged mice in vivo. Additionally, in aged liver nuclei NanoOrange revealed protein-dense foci that colocalize with centromeric heterochromatin.

  5. Kinematics of a former oceanic plate of the Neotethys revealed by deformation in the Ulukışla basin (Turkey)

    NARCIS (Netherlands)

    Gürer, Derya; van Hinsbergen, Douwe J J; Matenco, Liviu; Corfu, Fernando; Cascella, Antonio

    2016-01-01

    Kinematic reconstruction of modern ocean basins shows that since Pangea breakup a vast area in the Neotethyan realm was lost to subduction. Here we develop a first-order methodology to reconstruct the kinematic history of the lost plates of the Neotethys, using records of subducted plates accreted

  6. Genetic evidence for gonochoristic reproduction in gynogenetic silver crucian carp (Carassius auratus gibelio bloch) as revealed by RAPD assays.

    Science.gov (United States)

    Zhou, L; Wang, Y; Gui, J F

    2000-11-01

    Sex evolution has been a debating focus in evolutionary genetics. In lower vertebrates of reptiles, amphibians, and fish, a species or a bioform reproduces either sexually or asexually but never both. A few species were found to consist of all females in fish. These all-female species can propagate by asexual reproduction modes, such as gynogenesis and hybridogenesis. However, the coexistence of sexuality and asexuality in a single species was recently noted only in a cyprinid fish silver crucian carp, Carassius auratus gibelio. This fish had been demonstrated to be capable of gynogenesis stimulated by sperm from other related species. Surprisingly, natural populations of this fish consist of a minor but significant portion (approx. 20%) of males. As different clones with specific phenotypic and genetic characteristics have been found, and RAPD markers specific to each clone have recently been identified, this fish offers many advantages for analyzing whether or not genetic recombination occurs between different clones. In this study, artificial propagation was performed in clone F and clone D. Ovulated eggs from clone F were divided into two parts and respectively inseminated with sperm from a clone D male and from a red common carp (Cyprinus carpio) male. The control clone D individuals were selected from gynogenetic offspring of clone D activated by sperm of red common carp. The phenotype and sex ratio in the experimental groups were also observed. Using RAPD molecular markers, which allow for reliable discrimination and genetic analysis of different clones, we have revealed direct molecular evidence for gonochoristic reproduction in the gynogenetic silver crucian carp and confirmed a previous hypothesis that the silver crucian carp might reproduce both gynogenetically and gonochoristically. Therefore, we conclude that the silver crucian carp possesses two reproductive modes, i.e., gynogenetic and gonochoristic reproduction. The response mechanism of two

  7. 4D-CT scans reveal reduced magnitude of respiratory liver motion achieved by different abdominal compression plate positions in patients with intrahepatic tumors undergoing helical tomotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Yong, E-mail: hu.yong@zs-hospital.sh.cn; Zhou, Yong-Kang, E-mail: zhouyk2009@163.com; Chen, Yi-Xing, E-mail: chen.yixing@zs-hospital.sh.cn; Shi, Shi-Ming, E-mail: shiming32@126.com; Zeng, Zhao-Chong, E-mail: zeng.zhaochong@zs-hospital.sh.cn [Department of Radiation Oncology, Zhongshan Hospital, Fudan University, 180 Feng Lin Road, Shanghai 200032 (China)

    2016-07-15

    Purpose: While abdominal compression (AC) can be used to reduce respiratory liver motion in patients receiving helical tomotherapy for hepatocellular carcinoma, the nature and extent of this effect is not well described. The purpose of this study was to evaluate the changes in magnitude of three-dimensional liver motion with abdominal compression using four-dimensional (4D) computed tomography (CT) images of several plate positions. Methods: From January 2012 to October 2015, 72 patients with intrahepatic carcinoma and divided into four groups underwent 4D-CT scans to assess respiratory liver motion. Of the 72 patients, 19 underwent abdominal compression of the cephalic area between the subxiphoid and umbilicus (group A), 16 underwent abdominal compression of the caudal region between the subxiphoid area and the umbilicus (group B), 11 patients underwent abdominal compression of the caudal umbilicus (group C), and 26 patients remained free breathing (group D). 4D-CT images were sorted into ten-image series, according to the respiratory phase from the end inspiration to the end expiration, and then transferred to treatment planning software. All liver contours were drawn by a single physician and confirmed by a second physician. Liver relative coordinates were automatically generated to calculate the liver respiratory motion in different axial directions to compile the 10 ten contours into a single composite image. Differences in respiratory liver motion were assessed with a one-way analysis of variance test of significance. Results: The average respiratory liver motion in the Y axial direction was 4.53 ± 1.16, 7.56 ± 1.30, 9.95 ± 2.32, and 9.53 ± 2.62 mm in groups A, B, C, and D, respectively, with a significant change among the four groups (p < 0.001). Abdominal compression was most effective in group A (compression plate on the subxiphoid area), with liver displacement being 2.53 ± 0.93, 4.53 ± 1.16, and 2.14 ± 0.92 mm on the X-, Y-, and Z

  8. Cold plate

    Energy Technology Data Exchange (ETDEWEB)

    Marroquin, Christopher M.; O' Connell, Kevin M.; Schultz, Mark D.; Tian, Shurong

    2018-02-13

    A cold plate, an electronic assembly including a cold plate, and a method for forming a cold plate are provided. The cold plate includes an interface plate and an opposing plate that form a plenum. The cold plate includes a plurality of active areas arranged for alignment over respective heat generating portions of an electronic assembly, and non-active areas between the active areas. A cooling fluid flows through the plenum. The plenum, at the non-active areas, has a reduced width and/or reduced height relative to the plenum at the active areas. The reduced width and/or height of the plenum, and exterior dimensions of cold plate, at the non-active areas allow the non-active areas to flex to accommodate surface variations of the electronics assembly. The reduced width and/or height non-active areas can be specifically shaped to fit between physical features of the electronics assembly.

  9. Plating laboratory

    International Nuclear Information System (INIS)

    Seamster, A.G.; Weitkamp, W.G.

    1984-01-01

    The lead plating of the prototype resonator has been conducted entirely in the plating laboratory at SUNY Stony Brook. Because of the considerable cost and inconvenience in transporting personnel and materials to and from Stony Brook, it is clearly impractical to plate all the resonators there. Furthermore, the high-beta resonator cannot be accommodated at Stony Brook without modifying the set up there. Consequently the authors are constructing a plating lab in-house

  10. A novel transgenic mouse model of growth plate dysplasia reveals that decreased chondrocyte proliferation due to chronic ER stress is a key factor in reduced bone growth

    Directory of Open Access Journals (Sweden)

    Benedetta Gualeni

    2013-11-01

    Disease mechanisms leading to different forms of chondrodysplasia include extracellular matrix (ECM alterations and intracellular stress resulting in abnormal changes to chondrocyte proliferation and survival. Delineating the relative contribution of these two disease mechanisms is a major challenge in understanding disease pathophysiology in genetic skeletal diseases and a prerequisite for developing effective therapies. To determine the influence of intracellular stress and changes in chondrocyte phenotype to the development of chondrodysplasia, we targeted the expression of the G2320R mutant form of thyroglobulin to the endoplasmic reticulum (ER of resting and proliferating chondrocytes. Previous studies on this mutant protein have shown that it induces intracellular aggregates and causes cell stress and death in the thyroid gland. The expression and retention of this exogenous mutant protein in resting and proliferating chondrocytes resulted in a chronic cell stress response, growth plate dysplasia and reduced bone growth, without inducing any alterations to the architecture and organization of the cartilage ECM. More significantly, the decreased bone growth seemed to be the direct result of reduced chondrocyte proliferation in the proliferative zone of growth plates in transgenic mice, without transcriptional activation of a classical unfolded protein response (UPR or apoptosis. Overall, these data show that mutant protein retention in the ER of resting and proliferative zone chondrocytes is sufficient to cause disrupted bone growth. The specific disease pathways triggered by mutant protein retention do not necessarily involve a prototypic UPR, but all pathways impact upon chondrocyte proliferation in the cartilage growth plate.

  11. The fluorometric microculture cytotoxicity assay.

    Science.gov (United States)

    Lindhagen, Elin; Nygren, Peter; Larsson, Rolf

    2008-01-01

    The fluorometric microculture cytotoxicity assay (FMCA) is a nonclonogenic microplate-based cell viability assay used for measurement of the cytotoxic and/or cytostatic effect of different compounds in vitro. The assay is based on hydrolysis of the probe, fluorescein diacetate (FDA) by esterases in cells with intact plasma membranes. The assay is available as both a semiautomated 96-well plate setup and a 384-well plate version fully adaptable to robotics. Experimental plates are prepared with a small amount of drug solution and can be stored frozen. Cells are seeded on the plates and cell viability is evaluated after 72 h. The protocol described here is applicable both for cell lines and freshly prepared tumor cells from patients and is suitable both for screening in drug development and as a basis for a predictive test for individualization of anticancer drug therapy.

  12. Plate boundary deformation in North Iceland during 1992–2009 revealed by InSAR time-series analysis and GPS

    KAUST Repository

    Metzger, Sabrina

    2014-08-20

    In North Iceland, extensional plate motion is accommodated by the Northern Volcanic Zone, a set of en-echelon volcanic systems, and the Tjörnes Fracture Zone, a transform offset in the mid-Atlantic Ridge consisting of two parallel transform lineaments. The southern lineament, the Húsavík–Flatey fault, is a 100 km-long right-lateral strike slip fault that has not ruptured for more than 140 years and poses a significant seismic hazard to Húsavík, a fishing town located by the fault, and to other coastal communities. We present results of InSAR time-series analysis data spanning almost two decades (1992–2009) that show extensional and interseismic deformation within the Northern Volcanic Zone and the on-shore part of the Tjörnes Fracture Zone. The results also exhibit transient inflation at Theistareykir volcano, deflation at Krafla central volcano and a broad uplift north of Krafla. The current plate extension is not uniform across the Northern Volcanic Zone, but concentrated at the western fissures of the Theistareykir volcanic system and the outermost fissures of the Krafla fissure swarm. We combine a back-slip plate boundary model with a set of point pressure sources representing volcanic changes to describe the current extensional plate boundary deformation and update the previous estimations of the locking depth and slip rate of the Húsavík–Flatey fault that were based on GPS data alone. Using different combinations of input data, we find that the Húsavík–Flatey fault has a locking depth of 6–10 km and, with a slip rate of 6–9 mm/yr, is accommodating about a third of the full transform motion. We furthermore show that while the InSAR data provide important constraints on the volcanic deformation within the NVZ, they do not significantly improve the model parameter estimation for the HFF, as the dense GPS network appears to better capture the deformation across the fault.

  13. Plate boundary deformation in North Iceland during 1992–2009 revealed by InSAR time-series analysis and GPS

    KAUST Repository

    Metzger, Sabrina; Jonsson, Sigurjon

    2014-01-01

    In North Iceland, extensional plate motion is accommodated by the Northern Volcanic Zone, a set of en-echelon volcanic systems, and the Tjörnes Fracture Zone, a transform offset in the mid-Atlantic Ridge consisting of two parallel transform lineaments. The southern lineament, the Húsavík–Flatey fault, is a 100 km-long right-lateral strike slip fault that has not ruptured for more than 140 years and poses a significant seismic hazard to Húsavík, a fishing town located by the fault, and to other coastal communities. We present results of InSAR time-series analysis data spanning almost two decades (1992–2009) that show extensional and interseismic deformation within the Northern Volcanic Zone and the on-shore part of the Tjörnes Fracture Zone. The results also exhibit transient inflation at Theistareykir volcano, deflation at Krafla central volcano and a broad uplift north of Krafla. The current plate extension is not uniform across the Northern Volcanic Zone, but concentrated at the western fissures of the Theistareykir volcanic system and the outermost fissures of the Krafla fissure swarm. We combine a back-slip plate boundary model with a set of point pressure sources representing volcanic changes to describe the current extensional plate boundary deformation and update the previous estimations of the locking depth and slip rate of the Húsavík–Flatey fault that were based on GPS data alone. Using different combinations of input data, we find that the Húsavík–Flatey fault has a locking depth of 6–10 km and, with a slip rate of 6–9 mm/yr, is accommodating about a third of the full transform motion. We furthermore show that while the InSAR data provide important constraints on the volcanic deformation within the NVZ, they do not significantly improve the model parameter estimation for the HFF, as the dense GPS network appears to better capture the deformation across the fault.

  14. Quantifying rates of cell migration and cell proliferation in co-culture barrier assays reveals how skin and melanoma cells interact during melanoma spreading and invasion.

    Science.gov (United States)

    Haridas, Parvathi; Penington, Catherine J; McGovern, Jacqui A; McElwain, D L Sean; Simpson, Matthew J

    2017-06-21

    Malignant spreading involves the migration of cancer cells amongst other native cell types. For example, in vivo melanoma invasion involves individual melanoma cells migrating through native skin, which is composed of several distinct subpopulations of cells. Here, we aim to quantify how interactions between melanoma and fibroblast cells affect the collective spreading of a heterogeneous population of these cells in vitro. We perform a suite of circular barrier assays that includes: (i) monoculture assays with fibroblast cells; (ii) monoculture assays with SK-MEL-28 melanoma cells; and (iii) a series of co-culture assays initiated with three different ratios of SK-MEL-28 melanoma cells and fibroblast cells. Using immunostaining, detailed cell density histograms are constructed to illustrate how the two subpopulations of cells are spatially arranged within the spreading heterogeneous population. Calibrating the solution of a continuum partial differential equation to the experimental results from the monoculture assays allows us to estimate the cell diffusivity and the cell proliferation rate for the melanoma and the fibroblast cells, separately. Using the parameter estimates from the monoculture assays, we then make a prediction of the spatial spreading in the co-culture assays. Results show that the parameter estimates obtained from the monoculture assays lead to a reasonably accurate prediction of the spatial arrangement of the two subpopulations in the co-culture assays. Overall, the spatial pattern of spreading of the melanoma cells and the fibroblast cells is very similar in monoculture and co-culture conditions. Therefore, we find no clear evidence of any interactions other than cell-to-cell contact and crowding effects. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Comparative Analysis of Bone Structural Parameters Reveals Subchondral Cortical Plate Resorption and Increased Trabecular Bone Remodeling in Human Facet Joint Osteoarthritis

    Directory of Open Access Journals (Sweden)

    Cordula Netzer

    2018-03-01

    Full Text Available Facet joint osteoarthritis is a prominent feature of degenerative spine disorders, highly prevalent in ageing populations, and considered a major cause for chronic lower back pain. Since there is no targeted pharmacological therapy, clinical management of disease includes analgesic or surgical treatment. The specific cellular, molecular, and structural changes underpinning facet joint osteoarthritis remain largely elusive. The aim of this study was to determine osteoarthritis-related structural alterations in cortical and trabecular subchondral bone compartments. To this end, we conducted comparative micro computed tomography analysis in healthy (n = 15 and osteoarthritic (n = 22 lumbar facet joints. In osteoarthritic joints, subchondral cortical plate thickness and porosity were significantly reduced. The trabecular compartment displayed a 42 percent increase in bone volume fraction due to an increase in trabecular number, but not trabecular thickness. Bone structural alterations were associated with radiological osteoarthritis severity, mildly age-dependent but not gender-dependent. There was a lack of association between structural parameters of cortical and trabecular compartments in healthy and osteoarthritic specimens. The specific structural alterations suggest elevated subchondral bone resorption and turnover as a potential treatment target in facet joint osteoarthritis.

  16. Simulating the influence of plasma protein on measured receptor affinity in biochemical assays reveals the utility of Schild analysis for estimating compound affinity for plasma proteins.

    Science.gov (United States)

    Blakeley, D; Sykes, D A; Ensor, P; Bertran, E; Aston, P J; Charlton, S J

    2015-11-01

    Plasma protein binding (PPB) influences the free fraction of drug available to bind to its target and is therefore an important consideration in drug discovery. While traditional methods for assessing PPB (e.g. rapid equilibrium dialysis) are suitable for comparing compounds with relatively weak PPB, they are not able to accurately discriminate between highly bound compounds (typically >99.5%). The aim of the present work was to use mathematical modelling to explore the potential utility of receptor binding and cellular functional assays to estimate the affinity of compounds for plasma proteins. Plasma proteins are routinely added to in vitro assays, so a secondary goal was to investigate the effect of plasma proteins on observed ligand-receptor interactions. Using the principle of conservation of mass and the law of mass action, a cubic equation was derived describing the ligand-receptor complex [LR] in the presence of plasma protein at equilibrium. The model demonstrates the profound influence of PPB on in vitro assays and identifies the utility of Schild analysis, which is usually applied to determine receptor-antagonist affinities, for calculating affinity at plasma proteins (termed KP ). We have also extended this analysis to functional effects using operational modelling and demonstrate that these approaches can also be applied to cell-based assay systems. These mathematical models can potentially be used in conjunction with experimental data to estimate drug-plasma protein affinities in the earliest phases of drug discovery programmes. © 2015 The British Pharmacological Society.

  17. Safety Evaluation of Chinese Medicine Injections with a Cell Imaging-Based Multiparametric Assay Revealed a Critical Involvement of Mitochondrial Function in Hepatotoxicity

    Directory of Open Access Journals (Sweden)

    Meng Wang

    2015-01-01

    Full Text Available The safety of herbal medicine products has been a widespread concern due to their complex chemical nature and lack of proper evaluation methods. We have adapted a sensitive and reproducible multiparametric cell-based high-content analysis assay to evaluate the hepatic-safety of four Chinese medicine injections and validated it with classical animal-based toxicity assays. Our results suggested that the reported hepatotoxicity by one of the drugs, Fufangkushen injection, could be attributed at least in part to the interference of mitochondrial function in human HepG2 cells by some of its constituents. This method should be useful for both preclinical screen in a drug discovery program and postclinical evaluation of herbal medicine preparations.

  18. AKT1, LKB1, and YAP1 revealed as MYC interactors with NanoLuc-based protein-fragment complementation assay. | Office of Cancer Genomics

    Science.gov (United States)

    The c-Myc (MYC) transcription factor is a major cancer driver and a well-validated therapeutic target. However, directly targeting MYC has been challenging. Thus, identifying proteins that interact with and regulate MYC may provide alternative strategies to inhibit its oncogenic activity. Here we report the development of a NanoLuc®-based protein-fragment complementation assay (NanoPCA) and mapping of the MYC protein interaction hub in live mammalian cells.

  19. RAS - Screens & Assays - Drug Discovery

    Science.gov (United States)

    The RAS Drug Discovery group aims to develop assays that will reveal aspects of RAS biology upon which cancer cells depend. Successful assay formats are made available for high-throughput screening programs to yield potentially effective drug compounds.

  20. Nongenotoxic effects and a reduction of the DXR-induced genotoxic effects of Helianthus annuus Linné (sunflower) seeds revealed by micronucleus assays in mouse bone marrow.

    Science.gov (United States)

    Boriollo, Marcelo Fabiano Gomes; Souza, Luiz Silva; Resende, Marielly Reis; Silva, Thaísla Andrielle da; Oliveira, Nelma de Mello Silva; Resck, Maria Cristina Costa; Dias, Carlos Tadeu dos Santos; Fiorini, João Evangelista

    2014-04-02

    This research evaluated the genotoxicity of oil and tincture of H. annuus L. seeds using the micronucleus assay in bone marrow of mice. The interaction between these preparations and the genotoxic effects of doxorubicin (DXR) was also analysed (antigenotoxicity test). Experimental groups were evaluated at 24-48 h post treatment with N-Nitroso-N-ethylurea (positive control - NEU), DXR (chemotherapeutic), NaCl (negative control), a sunflower tincture (THALS) and two sources of sunflower oils (POHALS and FOHALS). Antigenotoxic assays were carried out using the sunflower tincture and oils separately and in combination with NUE or DXR. For THALS, analysis of the MNPCEs showed no significant differences between treatment doses (250-2,000 mg.Kg-1) and NaCl. A significant reduction in MNPCE was observed when THALS (2,000 mg.Kg-1) was administered in combination with DXR (5 mg.Kg-1). For POHALS or FOHALS, analysis of the MNPCEs also showed no significant differences between treatment doses (250-2,000 mg.Kg-1) and NaCl. However, the combination DXR + POHALS (2,000 mg.Kg-1) or DXR + FOHALS (2,000 mg.Kg-1) not contributed to the MNPCEs reduction. This research suggests absence of genotoxicity of THALS, dose-, time- and sex-independent, and its combination with DXR can reduce the genotoxic effects of DXR. POHALS and FOHALS also showed absence of genotoxicity, but their association with DXR showed no antigenotoxic effects.

  1. A modified anthrax toxin-based enzyme-linked immunospot assay reveals robust T cell responses in symptomatic and asymptomatic Ebola virus exposed individuals.

    Science.gov (United States)

    Herrera, Bobby Brooke; Hamel, Donald J; Oshun, Philip; Akinsola, Rolake; Akanmu, Alani S; Chang, Charlotte A; Eromon, Philomena; Folarin, Onikepe; Adeyemi, Kayode T; Happi, Christian T; Lu, Yichen; Ogunsola, Folasade; Kanki, Phyllis J

    2018-05-01

    Ebola virus (EBOV) caused more than 11,000 deaths during the 2013-2016 epidemic in West Africa without approved vaccines or immunotherapeutics. Despite its high lethality in some individuals, EBOV infection can produce little to no symptoms in others. A better understanding of the immune responses in individuals who experienced minimally symptomatic and asymptomatic infection could aid the development of more effective vaccines and antivirals against EBOV and related filoviruses. Between August and November 2017, blood samples were collected from 19 study participants in Lagos, Nigeria, including 3 Ebola virus disease (EVD) survivors, 10 individuals with documented close contact with symptomatic EVD patients, and 6 control healthcare workers for a cross-sectional serosurvey and T cell analysis. The Lagos samples, as well as archived serum collected from healthy individuals living in surrounding areas of the 1976 Democratic Republic of Congo (DRC) epidemic, were tested for EBOV IgG using commercial enzyme-linked immunosorbent assays (ELISAs) and Western blots. We detected antibodies in 3 out of 3 Lagos survivors and identified 2 seropositive individuals not known to have ever been infected. Of the DRC samples tested, we detected antibodies in 9 out of 71 (12.7%). To characterize the T cell responses in the Lagos samples, we developed an anthrax toxin-based enzyme-linked immunospot (ELISPOT) assay. The seropositive asymptomatic individuals had T cell responses against EBOV nucleoprotein, matrix protein, and glycoprotein 1 that were stronger in magnitude compared to the survivors. Our data provide further evidence of EBOV exposure in individuals without EVD-like illness and, for the first time, demonstrate that these individuals have T cell responses that are stronger in magnitude compared to severe cases. These findings suggest that T cell immunity may protect against severe EVD, which has important implications for vaccine development.

  2. Clonogenic assay: adherent cells.

    Science.gov (United States)

    Rafehi, Haloom; Orlowski, Christian; Georgiadis, George T; Ververis, Katherine; El-Osta, Assam; Karagiannis, Tom C

    2011-03-13

    The clonogenic (or colony forming) assay has been established for more than 50 years; the original paper describing the technique was published in 1956. Apart from documenting the method, the initial landmark study generated the first radiation-dose response curve for X-ray irradiated mammalian (HeLa) cells in culture. Basically, the clonogenic assay enables an assessment of the differences in reproductive viability (capacity of cells to produce progeny; i.e. a single cell to form a colony of 50 or more cells) between control untreated cells and cells that have undergone various treatments such as exposure to ionising radiation, various chemical compounds (e.g. cytotoxic agents) or in other cases genetic manipulation. The assay has become the most widely accepted technique in radiation biology and has been widely used for evaluating the radiation sensitivity of different cell lines. Further, the clonogenic assay is commonly used for monitoring the efficacy of radiation modifying compounds and for determining the effects of cytotoxic agents and other anti-cancer therapeutics on colony forming ability, in different cell lines. A typical clonogenic survival experiment using adherent cells lines involves three distinct components, 1) treatment of the cell monolayer in tissue culture flasks, 2) preparation of single cell suspensions and plating an appropriate number of cells in petri dishes and 3) fixing and staining colonies following a relevant incubation period, which could range from 1-3 weeks, depending on the cell line. Here we demonstrate the general procedure for performing the clonogenic assay with adherent cell lines with the use of an immortalized human keratinocyte cell line (FEP-1811). Also, our aims are to describe common features of clonogenic assays including calculation of the plating efficiency and survival fractions after exposure of cells to radiation, and to exemplify modification of radiation-response with the use of a natural antioxidant

  3. Plate tectonics

    Digital Repository Service at National Institute of Oceanography (India)

    Chaubey, A.K.

    's continental drift theory was later disproved, it was one of the first times that the idea of crustal movement had been introduced to the scientific community; and it has laid the groundwork for the development of modern plate tectonics. In the early... of the structure of the atom was to physical sciences and the theory of evolution was to the life sciences. Tectonics is the study of the forces within the Earth that give rise to continents, ocean basins, mountain ranges, earthquake belts and other large-scale...

  4. Create Your Plate

    Medline Plus

    Full Text Available ... Plate Share Create Your Plate ! Share: Seven Simple Steps to Create Your Plate It's simple and effective ... foods within each food category. Try these seven steps to get started: Using your dinner plate, put ...

  5. Trend of telomerase activity change during human iPSC self-renewal and differentiation revealed by a quartz crystal microbalance based assay

    Science.gov (United States)

    Zhou, Yitian; Zhou, Ping; Xin, Yinqiang; Wang, Jie; Zhu, Zhiqiang; Hu, Ji; Wei, Shicheng; Ma, Hongwei

    2014-11-01

    Telomerase plays an important role in governing the life span of cells for its capacity to extend telomeres. As high activity of telomerase has been found in stem cells and cancer cells specifically, various methods have been developed for the evaluation of telomerase activity. To overcome the time-consuming procedures and complicated manipulations of existing methods, we developed a novel method named Telomeric Repeat Elongation Assay based on Quartz crystal microbalance (TREAQ) to monitor telomerase activity during the self-renewal and differentiation of human induced pluripotent stem cells (hiPSCs). TREAQ results indicated hiPSCs possess invariable telomerase activity for 11 passages on Matrigel and a steady decline of telomerase activity when differentiated for different periods, which is confirmed with existing golden standard method. The pluripotency of hiPSCs during differentiation could be estimated through monitoring telomerase activity and compared with the expression levels of markers of pluripotency gene via quantitative real time PCR. Regular assessment for factors associated with pluripotency or stemness was expensive and requires excessive sample consuming, thus TREAQ could be a promising alternative technology for routine monitoring of telomerase activity and estimate the pluripotency of stem cells.

  6. A fluorescence-coupled assay for gamma aminobutyric acid (GABA reveals metabolic stress-induced modulation of GABA content in neuroendocrine cancer.

    Directory of Open Access Journals (Sweden)

    Joseph E Ippolito

    Full Text Available Pathways involved in the synthesis of the neurotransmitter gamma-aminobutyric acid (GABA have been implicated in the pathogenesis of high grade neuroendocrine (NE neoplasms as well as neoplasms from a non-NE lineage. Using The Cancer Genome Atlas, overexpression of the GABA synthetic enzyme, glutamate decarboxylase 1 (GAD1, was found to be associated with decreased disease free-survival in prostate adenocarcinoma and decreased overall survival in clear cell renal cell carcinomas. Furthermore, GAD1 was found to be expressed in castrate-resistant prostate cancer cell lines, but not androgen-responsive cell lines. Using a novel fluorescence-coupled enzymatic microplate assay for GABA mediated through reduction of resazurin in a prostate neuroendocrine carcinoma (PNEC cell line, acid microenvironment-induced stress increased GABA levels while alkaline microenvironment-induced stress decreased GABA through modulation of GAD1 and glutamine synthetase (GLUL activities. Moreover, glutamine but not glucose deprivation decreased GABA through modulation of GLUL. Consistent with evidence in prokaryotic and eukaryotic organisms that GABA synthesis mediated through GAD1 may play a crucial role in surviving stress, GABA may be an important mediator of stress survival in neoplasms. These findings identify GABA synthesis and metabolism as a potentially important pathway for regulating cancer cell stress response as well as a potential target for therapeutic strategies.

  7. In Vitro Endothelial Cell Proliferation Assay Reveals Distinct Levels of Proangiogenic Cytokines Characterizing Sera of Healthy Subjects and of Patients with Heart Failure

    Directory of Open Access Journals (Sweden)

    Rebecca Voltan

    2014-01-01

    Full Text Available Although myocardial angiogenesis is thought to play an important role in heart failure (HF, the involvement of circulating proinflammatory and proangiogenic cytokines in the pathogenesis and/or prognosis of HF has not been deeply investigated. By using a highly standardized proliferation assay with human endothelial cells, we first demonstrated that sera from older (mean age 52±7.6 years; n=46 healthy donors promoted endothelial cell proliferation to a significantly higher extent compared to sera obtained from younger healthy donors (mean age 29±8.6 years; n=20. The promotion of endothelial cell proliferation was accompanied by high serum levels of several proangiogenic cytokines. When we assessed endothelial cell proliferation in response to HF patients’ sera, we observed that a subset of sera (n=11 promoted cell proliferation to a significantly lesser extent compared to the majority of sera (n=18. Also, in this case, the difference between the patient groups in the ability to induce endothelial cell proliferation correlated to significant (P<0.05 differences in serum proangiogenic cytokine levels. Unexpectedly, HF patients associated to the highest endothelial proliferation index showed the worst prognosis as evaluated in terms of subsequent cardiovascular events in the follow-up, suggesting that high levels of circulating proangiogenic cytokines might be related to a worse prognosis.

  8. Immature intra-oceanic arc-type volcanism on the Izanagi Plate revealed by the geochemistry of the Daimaruyama greenstones in the Hiroo Complex, southern Hidaka Belt, central Hokkaido, Japan

    Science.gov (United States)

    Yamasaki, Toru; Nanayama, Futoshi

    2018-03-01

    results. Clinopyroxene trace element compositions indicate substantial enrichment of Ba in the magma, reflecting the participation of shallow subduction components such as aqueous fluids. Geochemical investigations reveal that the Daimaruyama greenstones were probably submarine volcanic rocks that formed as a result of the subduction of the Izanagi Plate within the Thalassa Ocean (the Thalassa Ocean was the eastern realm of Panthalassa, and represents the proto-Pacific Ocean) after the Early Cretaceous (Aptian-Albian; 125-101 Ma), and they were eventually accreted onto the Paleo-Kuril arc-trench system at 57-48 Ma to form an allochthonous block as part of the mélange facies of the Hiroo Complex on the landward slope of the trench.

  9. Elastic stability of thick auxetic plates

    International Nuclear Information System (INIS)

    Lim, Teik-Cheng

    2014-01-01

    Auxetic materials and structures exhibit a negative Poisson’s ratio while thick plates encounter shear deformation, which is not accounted for in classical plate theory. This paper investigates the effect of a negative Poisson’s ratio on thick plates that are subjected to buckling loads, taking into consideration the shear deformation using Mindlin plate theory. Using a highly accurate shear correction factor that allows for the effect of Poisson’s ratio, the elastic stability of circular and square plates are evaluated in terms of dimensionless parameters, namely the Mindlin-to-Kirchhoff critical buckling load ratio and Mindlin critical buckling load factors. Results for thick square plates reveal that both parameters increase as the Poisson’s ratio becomes more negative. In the case of thick circular plates, the Mindlin-to-Kirchhoff critical buckling load ratios and the Mindlin critical buckling load factors increase and decrease, respectively, as the Poisson’s ratio becomes more negative. The results obtained herein show that thick auxetic plates behave as thin conventional plates, and therefore suggest that the classical plate theory can be used to evaluate the elastic stability of thick plates if the Poisson’s ratio of the plate material is sufficiently negative. The results also suggest that materials with highly negative Poisson’s ratios are recommended for square plates, but not circular plates, that are subjected to buckling loads. (paper)

  10. Methods for identifying lipoxygenase producing microorganisms on agar plates

    NARCIS (Netherlands)

    Nyyssola, A.; Heshof, R.; Haarmann, T.; Eidner, J.; Westerholm-Parvinen, A.; Langfelder, K.; Kruus, K.; Graaff, de L.H.; Buchert, J.

    2012-01-01

    Plate assays for lipoxygenase producing microorganisms on agar plates have been developed. Both potassium iodide-starch and indamine dye formation methods were effective for detecting soybean lipoxygenase activity on agar plates. A positive result was also achieved using the beta-carotene bleaching

  11. Molecular assays reveal the presence of Theileria spp. and Babesia spp. in Asian water buffaloes (Bubalus bubalis, Linnaeus, 1758) in the Amazon region of Brazil.

    Science.gov (United States)

    Silveira, Júlia A G; de Oliveira, Cairo H S; Silvestre, Bruna T; Albernaz, Tatiana T; Leite, Rômulo C; Barbosa, José D; Oliveira, Carlos M C; Ribeiro, Múcio F B

    2016-07-01

    Approximately 50% of buffalo herds in Brazil are located in Pará state in northern Brazil. There are several properties where cattle and buffalo live and graze together, and thus, buffalo pathogens may threaten the health of cattle and vice versa. Therefore, knowledge of infectious agents of buffalo is essential for maintaining healthy livestock. Clinical disease caused by Theileria and Babesia parasites in the Asian water buffalo is not common, although these animals may act as reservoir hosts, and the detection of these hemoparasites in buffaloes is as important as it is in cattle. Studies of the infection of buffaloes by hemoparasites in Brazil are scarce. The objective of the present study was to investigate the occurrence of Piroplasmida parasites in Asian water buffaloes in the state of Pará in the Amazon region of Brazil using nested PCR assays and phylogenetic analysis. The 18S rRNA gene and ITS complete region were amplified from DNA extracted from blood samples collected from 308 apparently healthy buffaloes bred on six properties in the state of Pará, Brazil. The prevalence of positive buffalo samples was 4.2% (13/308) for Theileria spp., 3.6% (11/308) for Babesia bovis and 1% (3/308) for Babesia bigemina. Animals infected with Theileria were detected in 50% (3/6) of the assessed properties. Phylogenetic analyses indicated that the Theileria species detected in this study were closely related to Theileria buffeli, Theileria orientalis and Theileria sinensis. To our knowledge, this is the first report of Theileria in Asian water buffaloes in the Americas. The majority of Theileria-positive buffaloes (11/13) belong to a property that has a history of animals presenting lymphoproliferative disease of unknown etiology. Therefore, the present research suggests that this disorder can be associated with Theileria infection in this property. Our results provide new insights on the distribution and biological aspects of hemoparasites transmissible from

  12. Assessment of the repeatability and border-plate effects of the B158/B60 enzyme-linked-immunosorbent assay for the detection of circulating antigens (Ag-ELISA) of Taenia saginata.

    Science.gov (United States)

    Jansen, Famke; Dorny, Pierre; Berkvens, Dirk; Van Hul, Anke; Van den Broeck, Nick; Makay, Caroline; Praet, Nicolas; Gabriël, Sarah

    2016-08-30

    The monoclonal antibody-based circulating antigen detecting ELISA (B158/B60 Ag-ELISA) has been used elaborately in several studies for the diagnosis of human, bovine and porcine cysticercosis. Interpretation of test results requires a good knowledge of the test characteristics, including the repeatability and the effect of the borders of the ELISA plates. Repeatability was tested for 4 antigen-negative and 5 antigen-positive reference bovine serum samples by calculating the Percentage Coefficient of Variation (%CV) within and between plates, within and between runs, overall, for two batches of monoclonal antibodies and by 2 laboratory technicians. All CV values obtained were below 20% (except one: 24.45%), which indicates a good repeatability and a negligible technician error. The value of 24.45% for indicating the variability between batches of monoclonal antibodies for one positive sample is still acceptable for repeatability measures. Border effects were determined by calculating the %CV values between the inner and outer wells of one plate for 2 positive serum samples. Variability is a little more present in the outer wells but this effect is very small and no significant border effect was found. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Analysis of the asymmetrically expressed Ablim1 locus reveals existence of a lateral plate Nodal-independent left sided signal and an early, left-right independent role for nodal flow

    Directory of Open Access Journals (Sweden)

    Hilton Helen

    2010-05-01

    Full Text Available Abstract Background Vertebrates show clear asymmetry in left-right (L-R patterning of their organs and associated vasculature. During mammalian development a cilia driven leftwards flow of liquid leads to the left-sided expression of Nodal, which in turn activates asymmetric expression of the transcription factor Pitx2. While Pitx2 asymmetry drives many aspects of asymmetric morphogenesis, it is clear from published data that additional asymmetrically expressed loci must exist. Results A L-R expression screen identified the cytoskeletally-associated gene, actin binding lim protein 1 (Ablim1, as asymmetrically expressed in both the node and left lateral plate mesoderm (LPM. LPM expression closely mirrors that of Nodal. Significantly, Ablim1 LPM asymmetry was detected in the absence of detectable Nodal. In the node, Ablim1 was initially expressed symmetrically across the entire structure, resolving to give a peri-nodal ring at the headfold stage in a flow and Pkd2-dependent manner. The peri-nodal ring of Ablim1 expression became asymmetric by the mid-headfold stage, showing stronger right than left-sided expression. Node asymmetry became more apparent as development proceeded; expression retreated in an anticlockwise direction, disappearing first from the left anterior node. Indeed, at early somite stages Ablim1 shows a unique asymmetric expression pattern, in the left lateral plate and to the right side of the node. Conclusion Left LPM Ablim1 is expressed in the absence of detectable LPM Nodal, clearly revealing existence of a Pitx2 and Nodal-independent left-sided signal in mammals. At the node, a previously unrecognised action of early nodal flow and Pkd2 activity, within the pit of the node, influences gene expression in a symmetric manner. Subsequent Ablim1 expression in the peri-nodal ring reveals a very early indication of L-R asymmetry. Ablim1 expression analysis at the node acts as an indicator of nodal flow. Together these results make

  14. Laboratory assessment of anti-thrombotic therapy in heart failure, atrial fibrillation and coronary artery disease: insights using thrombelastography and a micro-titre plate assay of thrombogenesis and fibrinolysis.

    Science.gov (United States)

    Lau, Y C; Xiong, Q; Ranjit, P; Lip, G Y H; Blann, A D

    2016-08-01

    As heart failure, coronary artery disease and atrial fibrillation all bring a risk of thrombosis, anti-thrombotic therapy is recommended. Despite such treatment, major cardiovascular events such as myocardial infarction and stroke still occur, implying inadequate suppression of thrombus formation. Accordingly, identification of patients whose haemostasis remains unimpaired by treatment is valuable. We compared indices for assessing thrombogenesis and fibrinolysis by two different techniques in patients on different anti-thrombotic agents, i.e. aspirin or warfarin. We determined fibrin clot formation and fibrinolysis by a microplate assay and thromboelastography, and platelet marker soluble P selectin in 181 patients with acute or chronic heart failure, coronary artery disease who were taking either aspirin or warfarin. Five thromboelastograph indices and four microplate assay indices were different on aspirin versus warfarin (p indices rate of clot formation and rate of clot dissolution were independently related to aspirin or warfarin use (p ≤ 0.001). Five microplate assay indices, but no thrombelastograph index, were different (p indices were different (p ≤ 0.002) in warfarin users. The microplate assay indices of lag time and rate of clot formation were abnormal in chronic heart failure patients on aspirin, suggesting increased risk of thrombosis despite anti-platelet use. Soluble P selectin was lower in patients on aspirin (p = 0.0175) but failed to correlate with any other index of haemostasis. The microplate assay shows promise as a tool for dissecting thrombogenesis and fibrinolysis in cardiovascular disease, and the impact of antithrombotic therapy. Prospective studies are required to determine a role in predicting thrombotic risk.

  15. Development of a nematode offspring counting assay for rapid and simple soil toxicity assessment.

    Science.gov (United States)

    Kim, Shin Woong; Moon, Jongmin; Jeong, Seung-Woo; An, Youn-Joo

    2018-05-01

    Since the introduction of standardized nematode toxicity assays by the American Society for Testing and Materials (ASTM) and International Organization for Standardization (ISO), many studies have reported their use. Given that the currently used standardized nematode toxicity assays have certain limitations, in this study, we examined the use of a novel nematode offspring counting assay for evaluating soil ecotoxicity based on a previous soil-agar isolation method used to recover live adult nematodes. In this new assay, adult Caenorhabditis elegans were exposed to soil using a standardized toxicity assay procedure, and the resulting offspring in test soils attracted by a microbial food source in agar plates were counted. This method differs from previously used assays in terms of its endpoint, namely, the number of nematode offspring. The applicability of the bioassay was demonstrated using metal-spiked soils, which revealed metal concentration-dependent responses, and with 36 field soil samples characterized by different physicochemical properties and containing various metals. Principal component analysis revealed that texture fraction (clay, sand, and silt) and electrical conductivity values were the main factors influencing the nematode offspring counting assay, and these findings warrant further investigation. The nematode offspring counting assay is a rapid and simple process that can provide multi-directional toxicity assessment when used in conjunction with other standard methods. Copyright © 2018 Elsevier Ltd. All rights reserved.

  16. Transfer, imaging, and analysis plate for facile handling of 384 hanging drop 3D tissue spheroids.

    Science.gov (United States)

    Cavnar, Stephen P; Salomonsson, Emma; Luker, Kathryn E; Luker, Gary D; Takayama, Shuichi

    2014-04-01

    Three-dimensional culture systems bridge the experimental gap between in vivo and in vitro physiology. However, nonstandardized formation and limited downstream adaptability of 3D cultures have hindered mainstream adoption of these systems for biological applications, especially for low- and moderate-throughput assays commonly used in biomedical research. Here we build on our recent development of a 384-well hanging drop plate for spheroid culture to design a complementary spheroid transfer and imaging (TRIM) plate. The low-aspect ratio wells of the TRIM plate facilitated high-fidelity, user-independent, contact-based collection of hanging drop spheroids. Using the TRIM plate, we demonstrated several downstream analyses, including bulk tissue collection for flow cytometry, high-resolution low working-distance immersion imaging, and timely reagent delivery for enzymatic studies. Low working-distance multiphoton imaging revealed a cell type-dependent, macroscopic spheroid structure. Unlike ovarian cancer spheroids, which formed loose, disk-shaped spheroids, human mammary fibroblasts formed tight, spherical, and nutrient-limited spheroids. Beyond the applications we describe here, we expect the hanging drop spheroid plate and complementary TRIM plate to facilitate analyses of spheroids across the spectrum of throughput, particularly for bulk collection of spheroids and high-content imaging.

  17. Hormone assay

    International Nuclear Information System (INIS)

    Eisentraut, A.M.

    1977-01-01

    An improved radioimmunoassay is described for measuring total triiodothyronine or total thyroxine levels in a sample of serum containing free endogenous thyroid hormone and endogenous thyroid hormone bound to thyroid hormone binding protein. The thyroid hormone is released from the protein by adding hydrochloric acid to the serum. The pH of the separated thyroid hormone and thyroid hormone binding protein is raised in the absence of a blocking agent without interference from the endogenous protein. 125 I-labelled thyroid hormone and thyroid hormone antibodies are added to the mixture, allowing the labelled and unlabelled thyroid hormone and the thyroid hormone antibody to bind competitively. This results in free thyroid hormone being separated from antibody bound thyroid hormone and thus the unknown quantity of thyroid hormone may be determined. A thyroid hormone test assay kit is described for this radioimmunoassay. It provides a 'single tube' assay which does not require blocking agents for endogenous protein interference nor an external solid phase sorption step for the separation of bound and free hormone after the competitive binding step; it also requires a minimum number of manipulative steps. Examples of the assay are given to illustrate the reproducibility, linearity and specificity of the assay. (UK)

  18. Assay system

    International Nuclear Information System (INIS)

    Patzke, J.B.; Rosenberg, B.J.

    1984-01-01

    The accuracy of assays for monitoring concentrations of basic drugs in biological fluids containing a 1 -acid glycoproteins, such as blood (serum or plasma), is improved by the addition of certain organic phosphate compounds to minimize the ''protein effect.'' Kits containing the elements of the invention are also disclosed

  19. Radiation shielding plate

    International Nuclear Information System (INIS)

    Kobayashi, Torakichi; Sugawara, Takeo.

    1983-01-01

    Purpose: To reduce the weight and stabilize the configuration of a radiation shielding plate which is used in close contact with an object to be irradiated with radiation rays. Constitution: The radiation shielding plate comprises a substrate made of lead glass and a metallic lead coating on the surface of the substrate by means of plating, vapor deposition or the like. Apertures for permeating radiation rays are formed to the radiation shielding plate. Since the shielding plate is based on a lead glass plate, a sufficient mechanical strength can be obtained with a thinner structure as compared with the conventional plate made of metallic lead. Accordingly, if the shielding plate is disposed on a soft object to be irradiated with radiation rays, the object and the plate itself less deform to obtain a radiation irradiation pattern with distinct edges. (Moriyama, K.)

  20. Characterization of the commercially-available fluorescent chloroquine-BODIPY conjugate, LynxTag-CQGREEN, as a marker for chloroquine resistance and uptake in a 96-well plate assay.

    Directory of Open Access Journals (Sweden)

    Cheryl C Y Loh

    Full Text Available Chloroquine was a cheap, extremely effective drug against Plasmodium falciparum until resistance arose. One approach to reversing resistance is the inhibition of chloroquine efflux from its site of action, the parasite digestive vacuole. Chloroquine accumulation studies have traditionally relied on radiolabelled chloroquine, which poses several challenges. There is a need for development of a safe and biologically relevant substitute. We report here a commercially-available green fluorescent chloroquine-BODIPY conjugate, LynxTag-CQGREEN, as a proxy for chloroquine accumulation. This compound localized to the digestive vacuole of the parasite as observed under confocal microscopy, and inhibited growth of chloroquine-sensitive strain 3D7 more extensively than in the resistant strains 7G8 and K1. Microplate reader measurements indicated suppression of LynxTag-CQGREEN efflux after pretreatment of parasites with known reversal agents. Microsomes carrying either sensitive- or resistant-type PfCRT were assayed for uptake; resistant-type PfCRT exhibited increased accumulation of LynxTag-CQGREEN, which was suppressed by pretreatment with known chemosensitizers. Eight laboratory strains and twelve clinical isolates were sequenced for PfCRT and Pgh1 haplotypes previously reported to contribute to drug resistance, and pfmdr1 copy number and chloroquine IC50s were determined. These data were compared with LynxTag-CQGREEN uptake/fluorescence by multiple linear regression to identify genetic correlates of uptake. Uptake of the compound correlated with the logIC50 of chloroquine and, more weakly, a mutation in Pgh1, F1226Y.

  1. Create Your Plate

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    Full Text Available ... foods you want, but changes the portion sizes so you are getting larger portions of non-starchy ... plate. Then on one side, cut it again so you will have three sections on your plate. ...

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  6. Gentilly 2 divider plate replacement

    International Nuclear Information System (INIS)

    Forest, J.; Klisel, E.; McClellan, G.; Schnelder, W.

    1995-01-01

    The steam generators at the Gentilly 2 Nuclear Plant in operation since 1983 were built with primary divider plates of a bolted panel configuration. During a routine outage inspection, it was noted that two bolts had dislodged from the divider and were located lying in the primary head. Subsequent inspections revealed erosion damage to a substantial number of divider plate bolts and to a lesser extent, to the divider plate itself. After further inspection and repair the units were returned to operation, however, it was determined that a permanent replacement of the primary divider plates was going to be necessary. After evaluation of various options, it was decided that the panel type dividers would be replaced with a single piece floating design. The divider itself was to be of a one piece all-welded arrangement to be constructed from individual panels to be brought in through the manways. In view of the strength limitations of the bolted attachment of the upper seat bar to the tubesheet, a new welded seat bar was provided. To counteract erosion concerns, the new divider is fitted with erosion resistant inserts or weld buildup and with improved sealing features in order to minimize leakage and erosion. At an advanced stage in the design and manufacture of the components, the issue of divider strength during LOCA conditions came into focus. Analysis was performed to determine the strength and/or failure characteristics of the divider to a variety of small and large LOCA conditions. The paper describes the diagnosis of the original divider plates and the design, manufacture, field mobilization, installation and subsequent operation of the replacement divider plates. (author)

  7. Catalysts characteristics of Ni/YSZ core-shell according to plating conditions using electroless plating

    Science.gov (United States)

    Park, Hyun-Wook; Jang, Jae-Won; Lee, Young-Jin; Kim, Jin-Ho; Jeon, Dae-Woo; Lee, Jong-Heun; Hwang, Hae-jin; Lee, Mi-Jai

    2017-11-01

    This study aims to develop an anode catalyst for a solid oxide fuel cell (SOFC) using electroless nickel plating. We have proposed a new method for electroless plating of Ni metal on yttria-stabilized zirconia (YSZ) particles. We examine the uniformity of the Ni layer on the plated core-shell powder, in addition to the content of Ni and the reproducibility of the plating. We have also evaluated the carbon deposition rate and characteristics of the SOFC anode catalyst. To synthesize Ni-plated YSZ particles, the plated powder is heat-treated at 1200 °C. The resultant particles, which have an average size of 50 μm, were subsequently used in the experiment. The size of the Ni particles and the Ni content both increase with increasing plating temperature and plating time. The X-ray diffraction pattern reveals the growth of Ni particles. After heat-treatment, Ni is oxidized to NiO, leading to the co-existence of Ni and NiO; Ni3P is also observed due to the presence of phosphorous in the plating solution. Following heat treatment for 1 h at 1200 °C, Ni is mostly oxidized to NiO. The carbon deposition rate of the reference YSZ powder is 135%, while that of the Ni-plated YSZ is 1%-6%.

  8. Microfractionation revisited: a 1536 well high resolution screening assay

    NARCIS (Netherlands)

    Giera, M.A.; Heus, F.; Janssen, L.; Kool, J.; Lingeman, H.; Irth, H.

    2009-01-01

    The aim of the here presented study was to combine high performance liquid chromatography with plate reader technology in order to overcome certain drawbacks of integrated online systems as well as offline plate reader approaches. The described method combines an "at-line" enzyme assay for the

  9. Create Your Plate

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  11. An improved plating assay for determination of phage titer

    African Journals Online (AJOL)

    RACHEL

    antibiotics to control bacterial infections in swine (Thacker,. 2014). Phage therapy is re-valued by researchers to combat the growing menace of antibiotic-resistant infections (Torres-Barceló and Hochberg, 2016). Determination of phage titer in a sample is a key step in the study of the phage involved. It is very important to.

  12. A multiwell format assay for heparanase.

    Science.gov (United States)

    Behzad, Farhad; Brenchley, Paul E C

    2003-09-15

    This assay employs a biotinylated heparan sulfate glycosaminoglycan (HSGAG) substrate that is covalently linked to the surface of 96-well immunoassay plates. The ratio of biotin:HSGAG and the coating concentration of substrate bound to the wells have been optimized and allow removal of biotin HSGAG within 60 min of incubation at 37 degrees C in assay buffer with a standard dilution of bacterial heparitinase or platelet heparanase. Loss of biotin signal from the well surface is detected on incubation with peroxidase-streptavidin followed by color development using 3,3',5,5'-tetramethylbenzidine as the peroxidase substrate. The new assay allows specific detection of heparanase activity in multiple samples in a total time of 3 h including a 1-h substrate digestion step and is a significant improvement with regard to sensitivity, specificity, and ease of handling of multiple samples compared to other described assays. Heparanase specifically degrades the biotinylated HSGAG substrate, when used with an optimized assay buffer. A range of enzymes including collagenase, trypsin, plasmin, pepsin, chondroitinases, hyaluronidase, and neuraminidase show no effect on the substrate under optimized assay conditions. The covalent linkage of the substrate to the well prevents leaching of substrate and allows preparation and long-term storage of substrate-coated plates. The assay can be used to detect heparanase levels in clinical samples and cell culture supernatants and is ideal as a screening method for antagonists of enzyme activity.

  13. Create Your Plate

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  15. Create Your Plate

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  16. Create Your Plate

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    Full Text Available ... ready, you can try new foods within each food category. Try these seven steps to get started: Using your dinner plate, put a line down the middle of the plate. Then on one side, cut it ... and starchy foods. See this list of grains and starchy foods . ...

  17. Towards stacked zone plates

    International Nuclear Information System (INIS)

    Werner, S; Rehbein, S; Guttman, P; Heim, S; Schneider, G

    2009-01-01

    Fresnel zone plates are the key optical elements for soft and hard x-ray microscopy. For short exposure times and minimum radiation load of the specimen the diffraction efficiency of the zone plate objectives has to be maximized. As the efficiency strongly depends on the height of the diffracting zone structures the achievable aspect ratio of the nanostructures determines these limits. To reach aspect ratios ≥ 20:1 for high efficient optics we propose to superimpose zone plates on top of each other. With this multiplication approach the final aspect ratio is only limited by the number of stacked zone plate layers. For the stack process several nanostructuring process steps have to be developed and/or improved. Our results show for the first time two layers of zone plates stacked on top of each other.

  18. Soil plate bioassay: an effective method to determine ecotoxicological risks.

    Science.gov (United States)

    Boluda, R; Roca-Pérez, L; Marimón, L

    2011-06-01

    Heavy metals have become one of the most serious anthropogenic stressors for plants and other living organisms. Having efficient and feasible bioassays available to assess the ecotoxicological risks deriving from soil pollution is necessary. This work determines pollution by Cd, Co, Cr, Cu, Ni, Pb, V and Zn in two soils used for growing rice from the Albufera Natural Park in Valencia (Spain). Both were submitted to a different degree of anthropic activity, and their ecotoxicological risk was assessed by four ecotoxicity tests to compare their effectiveness: Microtox test, Zucconi test, pot bioassay (PB) and soil plate bioassay (SPB). The sensitivity of three plant species (barley, cress and lettuce) was also assessed. The results reveal a different degree of effectiveness and level of inhibition in the target organisms' growth depending on the test applied, to such an extent that the one-way analysis of variance showed significant differences only for the plate bioassay results, with considerable inhibition of root and shoot elongation in seedlings. Of the three plant species selected, lettuce was the most sensitive species to toxic effects, followed by cress and barley. Finally, the results also indicate that the SPB is an efficient, simple and economic alternative to other ecotoxicological assays to assess toxicity risks deriving from soil pollution. Copyright © 2011 Elsevier Ltd. All rights reserved.

  19. Anisotropic elastic plates

    CERN Document Server

    Hwu, Chyanbin

    2010-01-01

    As structural elements, anisotropic elastic plates find wide applications in modern technology. The plates here are considered to be subjected to not only in plane load but also transverse load. In other words, both plane and plate bending problems as well as the stretching-bending coupling problems are all explained in this book. In addition to the introduction of the theory of anisotropic elasticity, several important subjects have are discussed in this book such as interfaces, cracks, holes, inclusions, contact problems, piezoelectric materials, thermoelastic problems and boundary element a

  20. High loading uranium plate

    International Nuclear Information System (INIS)

    Wiencek, T.C.; Domagala, R.F.; Thresh, H.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 pari of diffusion bonded ductile metal cladding plates uniformly covering the meat, the meat hiving 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

  1. Heat insulating plates

    Energy Technology Data Exchange (ETDEWEB)

    Allan, J.A.F.

    1976-10-28

    Micro-porous insulation plates are dealt with, for example, how they are used in the insulation of heat storage devices. Since one side of such plates is exposed to a temperature of over 700/sup 0/C, a shrinkage of the glass texture of the covering can occur, which can exceed the shrinkage of the inner micro-porous material, so that cracks and splits in the high temperature side of the covering can come about. The task of the invention is to design the plate in such a way as to prevent this from happening. For this purpose the plate is provided, according to invention specifications, with flutes, waves, ribs, waffle or grid patterns and the covering is set into the recesses originating from this.

  2. Create Your Plate

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  4. Create Your Plate

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  5. Create Your Plate

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  17. Humvee Armor Plate Drilling

    National Research Council Canada - National Science Library

    2004-01-01

    When drilling holes in hard steel plate used in up-armor kits for Humvee light trucks, the Anniston Army Depot, Anniston, Alabama, requested the assistance of the National Center for Defense Manufacturing and Machining (NCDMM...

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  1. BAO Plate Archive Project

    Science.gov (United States)

    Mickaelian, A. M.; Gigoyan, K. S.; Gyulzadyan, M. V.; Paronyan, G. M.; Abrahamyan, H. V.; Andreasyan, H. R.; Azatyan, N. M.; Kostandyan, G. R.; Samsonyan, A. L.; Mikayelyan, G. A.; Farmanyan, S. V.; Harutyunyan, V. L.

    2017-12-01

    We present the Byurakan Astrophysical Observatory (BAO) Plate Archive Project that is aimed at digitization, extraction and analysis of archival data and building an electronic database and interactive sky map. BAO Plate Archive consists of 37,500 photographic plates and films, obtained with 2.6m telescope, 1m and 0.5m Schmidt telescopes and other smaller ones during 1947-1991. The famous Markarian Survey (or the First Byurakan Survey, FBS) 2000 plates were digitized in 2002-2005 and the Digitized FBS (DFBS, www.aras.am/Dfbs/dfbs.html) was created. New science projects have been conducted based on this low-dispersion spectroscopic material. Several other smaller digitization projects have been carried out as well, such as part of Second Byurakan Survey (SBS) plates, photographic chain plates in Coma, where the blazar ON 231 is located and 2.6m film spectra of FBS Blue Stellar Objects. However, most of the plates and films are not digitized. In 2015, we have started a project on the whole BAO Plate Archive digitization, creation of electronic database and its scientific usage. Armenian Virtual Observatory (ArVO, www.aras.am/Arvo/arvo.htm) database will accommodate all new data. The project runs in collaboration with the Armenian Institute of Informatics and Automation Problems (IIAP) and will continues during 4 years in 2015-2018. The final result will be an Electronic Database and online Interactive Sky map to be used for further research projects. ArVO will provide all standards and tools for efficient usage of the scientific output and its integration in international databases.

  2. Neutron imaging plates

    International Nuclear Information System (INIS)

    Niimura, Nobuo

    1995-01-01

    Imaging plates have been used in the field of medical diagnosis since long ago, but their usefulness was verified as the two-dimensional detector for analyzing the X-ray crystalline structure of high bio molecules like protein, and they have contributed to the remarkable progress in this field. The great contribution is due to the excellent features, such as the detection efficiency of about 100%, the positional resolution smaller than 0.2 mm, the dynamic range of five digits, and the area of several hundreds mm square. The neutron imaging plates have not yet obtained the sufficient results. It was planned to construct the neutron diffractometer for biological matters, and to put imaging plate neutron detectors (IP-ND) to practical use as the detector. The research on the development of IP-NDs was carried out, and the IPp-NDs having the performance comparable with that for X-ray were able to be produced. Imaging plates are the integral type two-dimensional radiation detector using photostimulated luminescence matters, and their principle is explained. As to neutron imaging plates, the converter, neutron detection efficiency and the flight of secondary particles in photo-stimulated luminescence matters are described. As for the present state of development of neutron imaging plates, the IP-NDs made for trial, the dynamic range, the positional resolution, the detection efficiency and the kinds of converters, and the application of IP-NDs are reported. (K.I.)

  3. Cadmium plating replacements

    Energy Technology Data Exchange (ETDEWEB)

    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.

  4. Bending and stretching of plates

    CERN Document Server

    Mansfield, E H; Hemp, W S

    1964-01-01

    The Bending and Stretching of Plates deals with elastic plate theory, particularly on small- and large-deflexion theory. Small-deflexion theory concerns derivation of basic equations, rectangular plates, plates of various shapes, plates whose boundaries are amenable to conformal transformation, plates with variable rigidity, and approximate methods. Large-deflexion theory includes general equations and some exact solutions, approximate methods in large-deflexion theory, asymptotic large-deflexion theories for very thin plates. Asymptotic theories covers membrane theory, tension field theory, a

  5. Analyzing the effects of size of hole on Plate failure

    Energy Technology Data Exchange (ETDEWEB)

    Behzad, Mohammadzadeh; Noh, Hyukchun [Sejong Univ., Seoul (Korea, Republic of)

    2013-05-15

    The load at critical point in which an infinitesimal increase in load can make the plate to buckle, is buckling load. When a plate element is subjected to direct compression, bending, shear, or a combination of these stresses in its plane, the plate may buckle locally before the member as a whole becomes unstable or before the yield stress of the material is reached. Holes can either increase or decrease critical load of a plate depending on its position and geometry. The presence of holes in plates will change the strength and stiffness, so the amounts of stress and its distribution which induce strain and buckling will be changed. This study deals with studying the buckling of plate with holes using finite element method(FEM). Buckling is one of the main reasons for steel members to fail during service life time. As plates are frequently used in the structures of nuclear power plants and in some cases making holes in plates is necessary, it is necessary to assay the capacity of the plates especially in terms of buckling. FEM is a useful approach which makes the plate analysis be performed with ease. This study relates the buckling load of plates with through-thickness holes to a dimensionless parameter (D/a). By increasing D/a ratio, the amount of plate strength is observed to be decreased. After D/a=0.5, the rate of decreasing is observed to be increased drastically. Therefore, it is better to use ratio D/a less than or equal to 0.5. As a further study, it is possible to investigate other aspects such as different thickness, different positions and so on.

  6. Analyzing the effects of size of hole on Plate failure

    International Nuclear Information System (INIS)

    Behzad, Mohammadzadeh; Noh, Hyukchun

    2013-01-01

    The load at critical point in which an infinitesimal increase in load can make the plate to buckle, is buckling load. When a plate element is subjected to direct compression, bending, shear, or a combination of these stresses in its plane, the plate may buckle locally before the member as a whole becomes unstable or before the yield stress of the material is reached. Holes can either increase or decrease critical load of a plate depending on its position and geometry. The presence of holes in plates will change the strength and stiffness, so the amounts of stress and its distribution which induce strain and buckling will be changed. This study deals with studying the buckling of plate with holes using finite element method(FEM). Buckling is one of the main reasons for steel members to fail during service life time. As plates are frequently used in the structures of nuclear power plants and in some cases making holes in plates is necessary, it is necessary to assay the capacity of the plates especially in terms of buckling. FEM is a useful approach which makes the plate analysis be performed with ease. This study relates the buckling load of plates with through-thickness holes to a dimensionless parameter (D/a). By increasing D/a ratio, the amount of plate strength is observed to be decreased. After D/a=0.5, the rate of decreasing is observed to be increased drastically. Therefore, it is better to use ratio D/a less than or equal to 0.5. As a further study, it is possible to investigate other aspects such as different thickness, different positions and so on

  7. Plating on Zircaloy-2

    International Nuclear Information System (INIS)

    Dini, J.W.; Johnson, H.R.; Jones, A.

    1979-03-01

    Zircaloy-2 is a difficult alloy to coat with an adherent electroplate because it easily forms a tenacious oxide film in air and aqueous solutions. Procedures reported in the literature and those developed at SLL for surmounting this problem were investigated. The best results were obtained when specimens were first etched in either an ammonium bifluoride/sulfuric acid or an ammonium bifluoride solution, plated, and then heated at 700 0 C for 1 hour in a constrained condition. Machining threads in the Zircaloy-2 for the purpose of providing sites for mechanical interlocking of the plating also proved satisfactory

  8. NICKEL PLATING PROCESS

    Science.gov (United States)

    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.

  9. Hydrodynamics of a flexible plate between pitching rigid plates

    Science.gov (United States)

    Kim, Junyoung; Kim, Daegyoum

    2017-11-01

    The dynamics of a flexible plate have been studied as a model problem in swimming and flying of animals and fluid-structure interaction of plants and flags. Motivated by fish schooling and an array of sea grasses, we investigate the dynamics of a flexible plate closely placed between two pitching rigid plates. In most studies on passive deformation of the flexible plate, the plate is immersed in a uniform flow or a wavy flow. However, in this study, the flexible plate experiences periodic deformation by the oscillatory flow generated by the prescribed pitching motion of the rigid plates. In our model, the pitching axes of the rigid plates and the clamping position of the flexible plate are aligned on the same line. The flexible plate shows various responses depending on length and pitching frequency of rigid plates, thickness of a flexible plate, and free-stream velocity. To find the effect of each variable on the response of the flexible plate, amplitude of a trailing edge and modal contribution of a flapping motion are compared, and flow structure around the flexible plate is examined.

  10. Imaging plates for nuclear radiations

    International Nuclear Information System (INIS)

    Abe, Ken; Takebe, Masahiro

    1997-01-01

    Full text. The imaging plate (IP, hereafter) is a new opto-electronic X-ray film developed by Fuji Photo Film Co. Ltd., formed with a large area of thin flexible plastic plate coated with photo-estimulable storage phosphor (e.g. Ba F Br: Eu 2+ ). Recently, it has been found highly sensitive to soft X-ray (SR), soft electrons, and also usual alpha, beta, gamma rays and others, e.g. cosmic rays, energy heavy ions, and moreover neutrons through suitable converters inside or outside of the IP. Many types of IP are now used in various fields, such as medical examinations, auto-radiography in vivo/ in situ/ in vitro, X-ray/neutron diffraction/ radiography, electron microscopy. RI contamination, assay of ore. The IP has other striking performances, e.e. extremely low intrinsic noises, a high position resolution, high detection efficiency (100-1000 times) as high as an X-ray film), extremely wide dynamic range of dose (more than 10 5 ). Besides the thermal fading yet left unresolved materially, the only feature lacking and that one has ben longing for is the radiation identification by itself. We found out that the IP has a full potential ability of radiation identification in itself. One evidence found is that the ratio of the twin peaks of the PSL (photo-stimulated luminescence) excitation spectra indicates simply the particle energies, studied and now established. Another is that the photo-beaching provides the fluorescent responses different enough to discriminate the radiations, yet in progress with cyclotron experiments, into the usage of double labeled bio tracers

  11. Create Your Plate

    Medline Plus

    Full Text Available ... meal-planning, . In this section Food Planning Meals Diabetes Meal Plans and a Healthy Diet Create Your Plate Meal Planning for Vegetarian Diets Gluten Free Diets Holiday Meal Planning Cook with Heart-Healthy Foods donate en -- A Future Without Diabetes - a-future-without-diabetes-2.html A Future ...

  12. Create Your Plate

    Medline Plus

    Full Text Available ... tool is not to scale because of the differences in types of vegetables. When creating your plate ... function (data) { $('#survey-errors').remove(); $('.survey-form .form-group .survey-alert-wrap').remove(); if (data.submitSurveyResponse.success == ' ...

  13. Create Your Plate

    Medline Plus

    Full Text Available ... Your Plate Meal Planning for Vegetarian Diets Gluten Free Diets Holiday Meal Planning Cook with Heart-Healthy Foods donate en -- A Future Without Diabetes - a-future-without-diabetes-2.html A Future Without Diabetes Donate towards research today and your gift will be matched. Donate ...

  14. Create Your Plate

    Medline Plus

    Full Text Available ... Planning Meals > Create Your Plate Share: Print Page Text Size: A A A Listen En Español Create ... Type 2 Education Series Hear audio clips and full recordings of past Q&A events at your ...

  15. Create Your Plate

    Medline Plus

    Full Text Available ... 1 Type 2 About Us Online Community Meal Planning Sign In Search: Search More Sites Search ≡ Are ... Fitness Home Food MyFoodAdvisor Recipes Association Cookbook Recipes Planning Meals Diabetes Meal Plans Create Your Plate Gluten ...

  16. Create Your Plate

    Medline Plus

    Full Text Available ... tax-deductible gift today can fund critical diabetes research and support vital diabetes education services that improve the ... effective way to manage your blood glucose levels and lose weight. With this method, you fill your plate with more non-starchy ...

  17. Microchannel plate photodetectors

    International Nuclear Information System (INIS)

    Majka, R.

    1977-01-01

    A review is given the status of development work on photodetectors using microchannel plates (MCP) as the electron gain element. Projections are made and opinions are presented on what might be available in the next few years. Several uses for these devices at ISABELLE are mentioned

  18. Parallel plate detectors

    International Nuclear Information System (INIS)

    Gardes, D.; Volkov, P.

    1981-01-01

    A 5x3cm 2 (timing only) and a 15x5cm 2 (timing and position) parallel plate avalanche counters (PPAC) are considered. The theory of operation and timing resolution is given. The measurement set-up and the curves of experimental results illustrate the possibilities of the two counters [fr

  19. Flat plate collector. Solarflachkollektor

    Energy Technology Data Exchange (ETDEWEB)

    Raab, N

    1979-03-29

    The invention refers to a flat solar collector with an absorber plate, which is arranged on a support and is covered by a transparent window, between which and the plate there is an air space. The previously known structures of this type had the disadvantage that the thermal expansion of the enclosed air caused considerable difficulties. The purpose of the invention is therefore to create a collector, which can be used on the modular system, retains its properties and is safe in spite of the great temperature variations. According to the invention this problem is solved by providing a compensating space in the collector, which is separated by a diaphragm from the airspace between the plate and the covering window. The airspace therefore remains sealed against the atmosphere, so that no dirt, corrosion of the inside and no condensation can reduce the efficiency of the collector. A rise in pressure due to an increase in temperature is immediately reduced by expansion of the diaphragm, which enters the compensation space. In order to increase the pressure in the airspace above the plate for increases in temperature, the compensation space is connected to the atmosphere. The diaphragm can be mirrored on the side towards the absorber, which makes the diaphragm into an insulating element, as it reflects radiated heat from the absorber.

  20. Structure of colloidal sphere-plate mixtures

    International Nuclear Information System (INIS)

    Doshi, N; Cinacchi, G; Van Duijneveldt, J S; Cosgrove, T; Prescott, S W; Grillo, I; Phipps, J; Gittins, D I

    2011-01-01

    In addition to containing spherical pigment particles, coatings usually contain plate-like clay particles. It is thought that these improve the opacity of the paint film by providing an efficient spacing of the pigment particles. This observation is counterintuitive, as suspensions of particles of different shapes and sizes tend to phase separate on increase of concentration. In order to clarify this matter a model colloidal system is studied here, with a sphere-plate diameter ratio similar to that found in paints. For dilute suspensions, small angle neutron scattering revealed that the addition of plates leads to enhanced density fluctuations of the spheres, in agreement with new theoretical predictions. On increasing the total colloid concentration the plates and spheres phase separate due to the disparity in their shape. This is in agreement with previous theoretical and experimental work on colloidal sphere-plate mixtures, where one particle acts as a depleting agent. The fact that no large scale phase separation is observed in coatings is ascribed to dynamic arrest in intimately mixed, or possibly micro-phase separated structures, at elevated concentration.

  1. Structure of colloidal sphere-plate mixtures

    Energy Technology Data Exchange (ETDEWEB)

    Doshi, N; Cinacchi, G; Van Duijneveldt, J S; Cosgrove, T; Prescott, S W [School of Chemistry, University of Bristol, Bristol BS8 1TS (United Kingdom); Grillo, I [Institut Laue-Langevin, 6 rue Jules Horowitz BP 156, 38042 Grenoble Cedex 9 (France); Phipps, J [Imerys Minerals Ltd, Par Moor Centre, Par Moor Road, Par, Cornwall PL24 2SQ (United Kingdom); Gittins, D I, E-mail: Giorgio.Cinacchi@bristol.ac.uk, E-mail: J.S.van-Duijneveldt@bristol.ac.uk [Imerys Performance and Filtration Minerals Ltd, 130 Castilian Drive, Goleta, CA 93117 (United States)

    2011-05-18

    In addition to containing spherical pigment particles, coatings usually contain plate-like clay particles. It is thought that these improve the opacity of the paint film by providing an efficient spacing of the pigment particles. This observation is counterintuitive, as suspensions of particles of different shapes and sizes tend to phase separate on increase of concentration. In order to clarify this matter a model colloidal system is studied here, with a sphere-plate diameter ratio similar to that found in paints. For dilute suspensions, small angle neutron scattering revealed that the addition of plates leads to enhanced density fluctuations of the spheres, in agreement with new theoretical predictions. On increasing the total colloid concentration the plates and spheres phase separate due to the disparity in their shape. This is in agreement with previous theoretical and experimental work on colloidal sphere-plate mixtures, where one particle acts as a depleting agent. The fact that no large scale phase separation is observed in coatings is ascribed to dynamic arrest in intimately mixed, or possibly micro-phase separated structures, at elevated concentration.

  2. Gentilly-2 and Point Lepreau divider plate replacement

    International Nuclear Information System (INIS)

    Schneider, W.; McClellan, G.; Weston, S.

    1996-01-01

    The steam generators at Hydro Quebec's Gentilly-2 and New Brunswick Power's Point Lepreau Nuclear Plants have been in operation since 1983, and were built with primary divider plates of a bolted panel configuration. During a routine outage inspection at Gentilly-2, it was noted that two bolts had dislodged from the divider plate and were located lying in the primary head. Subsequent inspections revealed erosion damage to a a substantial number of divider plate bolts and to a lesser extent, to the divider plate itself. After further inspection and repair the units were returned to operation, however, it was determined that a permanent replacement of the primary divider plates was going to be necessary. Upon evaluation of various options, it was decided that the panel type divider plates would be replaced with a single piece floating design. The divider plate itself was to be of one piece all-welded arrangement to be constructed from individual panels to be brought in through the manways. In view of the strength limitations of the bolted attachment of the upper seat bar to the tubesheet, a new welded seat was was provided. To counteract erosion concerns, the new divider plate is fitted with erosion resistant inserts of weld buildup and with improved sealing features in order to minimize leakage and erosion. At an advanced stage in the design and manufacture of the components, the issue of divider plate strength during loss of coolant accident (LOCA) conditions came into focus. Analysis was performed to determine the strength and/or failure characteristics of the divider plate to a variety of small and large LOCA conditions. Subsequently, Point Lepreau decided to replace their divider plates to address LOCA concerns. The paper describes the diagnosis of the original divider plates and the design. manufacture, field mobilization, installation and subsequent operation of the replacement divider plates. (author)

  3. Plate Full of Color

    Centers for Disease Control (CDC) Podcasts

    The Eagle Books are a series of four books that are brought to life by wise animal characters - Mr. Eagle, Miss Rabbit, and Coyote - who engage Rain That Dances and his young friends in the joy of physical activity, eating healthy foods, and learning from their elders about health and diabetes prevention. Plate Full of Color teaches the value of eating a variety of colorful and healthy foods.

  4. Effect of plate shapes in orifice plate type flowmeters

    International Nuclear Information System (INIS)

    Moeller, S.V.

    1984-01-01

    The study of unusual plate shapes in orifice plate type flowmeters is presented, with a view to providing data for the substitution of the plate with one centered circular orifice in those applications where its use is not possible. For this purpose, six pairs of plates with different forms, with and without chamfered edges, were made and tested in a closed water loop. Results show that, generally, the use of chamfers improves the results and, in the case of perforated and slotlike orificed plates, the narrow-ness of the fluid passage tends to make unnecessary its use. (Author) [pt

  5. Plate removal following orthognathic surgery.

    Science.gov (United States)

    Little, Mhairi; Langford, Richard Julian; Bhanji, Adam; Farr, David

    2015-11-01

    The objectives of this study are to determine the removal rates of orthognathic plates used during orthognathic surgery at James Cook University Hospital and describe the reasons for plate removal. 202 consecutive orthognathic cases were identified between July 2004 and July 2012. Demographics and procedure details were collected for these patients. Patients from this group who returned to theatre for plate removal between July 2004 and November 2012 were identified and their notes were analysed for data including reason for plate removal, age, smoking status, sex and time to plate removal. 3.2% of plates were removed with proportionally more plates removed from the mandible than the maxilla. 10.4% of patients required removal of one or more plate. Most plates were removed within the first post-operative year. The commonest reasons for plate removal were plate exposure and infection. The plate removal rates in our study are comparable to those seen in the literature. Copyright © 2015 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.

  6. Plate Tearing by a Cone

    DEFF Research Database (Denmark)

    Simonsen, Bo Cerup

    1997-01-01

    The present paper is concerned with steady-state plate tearing by a cone. This is a scenario where a cone is forced through a ductile metal plate with a constant lateral tip penetration in a motion in the plane of the plate. The considered process could be an idealisaton of the damage, which...... as for the out-of-plane reaction force....

  7. Normal and abnormal growth plate

    International Nuclear Information System (INIS)

    Kumar, R.; Madewell, J.E.; Swischuk, L.E.

    1987-01-01

    Skeletal growth is a dynamic process. A knowledge of the structure and function of the normal growth plate is essential in order to understand the pathophysiology of abnormal skeletal growth in various diseases. In this well-illustrated article, the authors provide a radiographic classification of abnormal growth plates and discuss mechanisms that lead to growth plate abnormalities

  8. Multiplexing a high-throughput liability assay to leverage efficiencies.

    Science.gov (United States)

    Herbst, John; Anthony, Monique; Stewart, Jeremy; Connors, David; Chen, Taosheng; Banks, Martyn; Petrillo, Edward W; Agler, Michele

    2009-06-01

    In order to identify potential cytochrome P-450 3A4 (drug-metabolizing enzyme) inducers at an early stage of the drug discovery process, a cell-based transactivation high-throughput luciferase reporter assay for the human pregnane X receptor (PXR) in HepG2 cells has been implemented and multiplexed with a viability end point for data interpretation, as part of a Lead Profiling portfolio of assays. As a routine part of Lead Profiling operations, assays are periodically evaluated for utility as well as for potential improvements in technology or process. We used a recent evaluation of our PXR-transactivation assay as a model for the application of Lean Thinking-based process analysis to lab-bench assay optimization and automation. This resulted in the development of a 384-well multiplexed homogeneous assay simultaneously detecting PXR transactivation and HepG2 cell cytotoxicity. In order to multiplex fluorescent and luminescent read-outs, modifications to each assay were necessary, which included optimization of multiple assay parameters such as cell density, plate type, and reagent concentrations. Subsequently, a set of compounds including known cytotoxic compounds and PXR inducers were used to validate the multiplexed assay. Results from the multiplexed assay correlate well with those from the singleplexed assay formats measuring PXR transactivation and viability separately. Implementation of the multiplexed assay for routine compound profiling provides improved data quality, sample conservation, cost savings, and resource efficiencies.

  9. From Plate Tectonic to Continental Dynamics

    Science.gov (United States)

    Molnar, P. H.

    2017-12-01

    By the early 1970s, the basics of plate tectonics were known. Although much understanding remained to be gained, as a topic of research, plate tectonics no longer defined the forefront of earth science. Not only had it become a foundation on which to build, but also the methods used to reveal it became tools to take in new directions. For me as a seismologist studying earthquakes and active processes, the deformation of continents offered an obvious topic to pursue. Obviously examining the deformation of continents and ignoring the widespread geologic evidence of both ongoing and finite deformation of crust would be stupid. I was blessed with the opportunity to learn from and collaborate with two of the best, Paul Tapponnier and Clark Burchfiel. Continental deformation differed from plate tectonics both because deformation was widespread but more importantly because crust shortens (extends) horizontally and thickens (thins), processes that can be ignored where plate tectonics - the relative motion of rigid plates - occurs. Where a plate boundary passes into a continent, not only must the forces that move plates do work against friction or other dissipative processes, but where high terrain is created, they must also do work against gravity, to create gravitational potential energy in high terrain. Peter Bird and Kenneth Piper and Philip England and Dan McKenzie showed that a two-dimensional thin viscous sheet with vertically averaged properties enabled both sources of resistance to be included without introducing excessive complexity and to be scaled by one dimensionless number, what the latter pair called the Argand number. Increasingly over the past thirty years, emphasis has shifted toward the role played by the mantle lithosphere, because of both its likely strength and its negative buoyancy, which makes it gravitationally unstable. Despite progress since realizing that rigid plates (the essence of plate tectonics) provides a poor description of continental

  10. Microbead agglutination based assays

    KAUST Repository

    Kodzius, Rimantas

    2013-01-21

    We report a simple and rapid room temperature assay for point-of-care (POC) testing that is based on specific agglutination. Agglutination tests are based on aggregation of microbeads in the presence of a specific analyte thus enabling the macroscopic observation. Such tests are most often used to explore antibody-antigen reactions. Agglutination has been used for protein assays using a biotin/streptavidin system as well as a hybridization based assay. The agglutination systems are prone to selftermination of the linking analyte, prone to active site saturation and loss of agglomeration at high analyte concentrations. We investigated the molecular target/ligand interaction, explaining the common agglutination problems related to analyte self-termination, linkage of the analyte to the same bead instead of different microbeads. We classified the agglutination process into three kinds of assays: a two- component assay, a three-component assay and a stepped three- component assay. Although we compared these three kinds of assays for recognizing DNA and protein molecules, the assay can be used for virtually any molecule, including ions and metabolites. In total, the optimized assay permits detecting analytes with high sensitivity in a short time, 5 min, at room temperature. Such a system is appropriate for POC testing.

  11. Transfer plate radioassay using adsorbed anti-insulin antibody to detect insulin secreted by islet cell cultures

    International Nuclear Information System (INIS)

    Scearce, R.M.; Oie, H.K.; Gazdar, A.F.; Chick, W.L.; Eisenbarth, G.S.

    1981-01-01

    A solid-phase radioimmunoassay for detection of insulin synthesized by islet cell clones is described. This assay employs anti-insulin antibody adsorbed onto fenestrated polyvinyl chloride 96-well plates ('transfer plates'). The calibrated aperture in the bottom of each transfer plate well permits fluid to enter the wells when transfer plates are lowered into microculture wells containing insulin. With this assay it is possible to rapidly screen hundreds of islet cell cultures for insulin production. The authors have used this assay to facilitate cloning of the RIN rat insulinoma cell line. The assay readily detects insulin synthesis by RIN cells and [ 125 I]insulin is not displaced by culture medium from cells which do not produce insulin. The transfer plate format should be applicable to semiautomate other radioimmunoassays. (Auth.)

  12. A source of artifact in the lacZ reversion assay in Escherichia coli.

    Science.gov (United States)

    Hoffmann, George R; Gray, Carol L; Lange, Paulina B; Marando, Christie I

    2015-06-01

    The lacZ reversion assay in Escherichia coli measures point mutations that occur by specific base substitutions and frameshift mutations. The tester strains cannot use lactose as a carbon source (Lac(-)), and revertants are easily detected by growth on lactose medium (Lac(+)). Six strains identify the six possible base substitutions, and five strains measure +G, -G, -CG, +A and -A frameshifts. Strong mutagens give dose-dependent increases in numbers of revertants per plate and revertant frequencies. Testing compounds that are arguably nonmutagens or weakly mutagenic, we often noted statistically significant dose-dependent increases in revertant frequency that were not accompanied by an absolute increase in numbers of revertants. The increase in frequency was wholly ascribable to a declining number of viable cells owing to toxicity. Analysis of the conditions revealed that the frequency of spontaneous revertants is higher when there are fewer viable cells per plate. The phenomenon resembles "adaptive" or "stress" mutagenesis, whereby lactose revertants accumulate in Lac(-) bacteria under starvation conditions in the absence of catabolite repression. Adaptive mutation is observed after long incubation and might be expected to be irrelevant in a standard assay using 48-h incubation. However, we found that elevated revertant frequencies occur under typical assay conditions when the bacterial lawn is thin, and this can cause increases in revertant frequency that mimic chemical mutagenesis when treatments are toxic but not mutagenic. Responses that resemble chemical mutagenesis were observed in the absence of mutagenic treatment in strains that revert by different frameshift mutations. The magnitude of the artifact is affected by cell density, dilution, culture age, incubation time, catabolite repression and the age and composition of media. Although the specific reversion assay is effective for quickly distinguishing classes of mutations induced by potent mutagens, its

  13. Analysis of a complex shape chain plate using Transmission Photoelasticity

    Directory of Open Access Journals (Sweden)

    Dasari N.

    2010-06-01

    -3]. A loading fixture was used to load the model during stress freezing in the furnace. The integrated fringe pattern of the offset chain plate is shown in Figure 2. Slices are cut in the offset zone to understand the nature of the stress distribution. A Simple photoelastic analysis of this has revealed a wealth of information to take suitable decision on the design of such plates taking into account the constraints in manufacturing.

  14. Study of uranium plating measurement

    International Nuclear Information System (INIS)

    Lin Jufang; Wen Zhongwei; Wang Mei; Wang Dalun; Liu Rong; Jiang Li; Lu Xinxin

    2007-06-01

    In neutron physics experiments, the measurement for plate-thickness of uranium can directly affect uncertainties of experiment results. To measure the plate-thickness of transform target (enriched uranium plating and depleted uranium plating), the back to back ionization chamber, small solid angle device and Au-Si surface barrier semi-conductor, were used in the experiment study. Also, the uncertainties in the experiment were analyzed. Because the inhomo-geneous of uranium lay of plate can quantitively affect the result, the homogeneity of uranium lay is checked, the experiment result reflects the homogeneity of uranium lay is good. (authors)

  15. Radioreceptor opioid assay

    International Nuclear Information System (INIS)

    Miller, R.J.; Chang, K.-J.

    1981-01-01

    A radioreceptor assay is described for assaying opioid drugs in biological fluids. The method enables the assay of total opioid activity, being specific for opioids as a class but lacking specificity within the class. A radio-iodinated opioid and the liquid test sample are incubated with an opiate receptor material. The percentage inhibition of the binding of the radio-iodinated compound to the opiate receptor is calculated and the opioid activity of the test liquid determined from a standard curve. Examples of preparing radio-iodinated opioids and assaying opioid activity are given. A test kit for the assay is described. Compared to other methods, this assay is cheap, easy and rapid. (U.K.)

  16. h5-HT1B receptor-mediated constitutive Gαi3-protein activation in stably transfected Chinese hamster ovary cells: an antibody capture assay reveals protean efficacy of 5-HT

    Science.gov (United States)

    Newman-Tancredi, Adrian; Cussac, Didier; Marini, Laetitia; Touzard, Manuelle; Millan, Mark J

    2003-01-01

    Serotonin 5-HT1B receptors couple to G-proteins of the Gi/o family. However, their activation of specific G-protein subtypes is poorly characterised. Using an innovative antibody capture/guanosine-5′-0-(3-[35S]thio)-triphosphate ([35S]GTPγS) binding strategy, we characterised Gαi3 subunit activation by h5-HT1B receptors stably expressed in Chinese hamster ovary (CHO) cells. The agonists, 5-HT, alniditan and BMS181,101, stimulated Gαi3, whereas methiothepin and SB224,289 behaved as inverse agonists. The selective 5-HT1B receptor ligand, S18127, modestly stimulated Gαi3 and reversed the actions of both 5-HT and methiothepin. S18127 (1 μM) also produced parallel, dextral shifts of the 5-HT and methiothepin isotherms. Isotopic dilution experiments ([35S]GTPγS versus GTPγS) revealed high-affinity [35S]GTPγS binding to Gαi3 subunits in the absence of receptor ligands indicating constitutive activity. High-affinity [35S]GTPγS binding was increased 2.8-fold by 5-HT with an increase in the affinity of GTPγS for Gαi3 subunits. In contrast, methiothepin halved the number of high-affinity binding sites and decreased their affinity. h5-HT1B receptor-mediated Gαi3 subunit activation was dependent on the concentration of NaCl. At 300 mM, 5-HT stimulated [35S]GTPγS binding, basal Gαi3 activation was low and methiothepin was inactive. In contrast, at 10 mM NaCl, basal activity was enhanced and the inverse agonist activity of methiothepin was accentuated. Under these conditions, 5-HT decreased Gαi3 activation. In conclusion, at h5-HT1B receptors expressed in CHO cells: (i) inverse agonist induced inhibition of Gαi3, and its reversal by S18127, reveals constitutive activation of this Gα subunit; (ii) constitutive Gαi3 activation can be quantified by isotopic dilution [35S]GTPγS binding and (iii) decreasing NaCl concentrations enhances Gαi3 activation and leads to protean agonist properties of 5-HT: that is a switch to inhibition of Gαi3. PMID:12684263

  17. h5-HT(1B) receptor-mediated constitutive Galphai3-protein activation in stably transfected Chinese hamster ovary cells: an antibody capture assay reveals protean efficacy of 5-HT.

    Science.gov (United States)

    Newman-Tancredi, Adrian; Cussac, Didier; Marini, Laetitia; Touzard, Manuelle; Millan, Mark J

    2003-03-01

    1. Serotonin 5-HT(1B) receptors couple to G-proteins of the Gi/o family. However, their activation of specific G-protein subtypes is poorly characterised. Using an innovative antibody capture/guanosine-5'-0-(3-[(35)S]thio)-triphosphate ([(35)S]GTPgammaS) binding strategy, we characterised Galpha(i3) subunit activation by h5-HT(1B) receptors stably expressed in Chinese hamster ovary (CHO) cells. 2. The agonists, 5-HT, alniditan and BMS181,101, stimulated Galpha(i3), whereas methiothepin and SB224,289 behaved as inverse agonists. The selective 5-HT(1B) receptor ligand, S18127, modestly stimulated Galpha(i3) and reversed the actions of both 5-HT and methiothepin. S18127 (1 micro M) also produced parallel, dextral shifts of the 5-HT and methiothepin isotherms. 3. Isotopic dilution experiments ([(35)S]GTPgammaS versus GTPgammaS) revealed high-affinity [(35)S]GTPgammaS binding to Galpha(i3) subunits in the absence of receptor ligands indicating constitutive activity. High-affinity [(35)S]GTPgammaS binding was increased 2.8-fold by 5-HT with an increase in the affinity of GTPgammaS for Galpha(i3) subunits. In contrast, methiothepin halved the number of high-affinity binding sites and decreased their affinity. 4. h5-HT(1B) receptor-mediated Galpha(i3) subunit activation was dependent on the concentration of NaCl. At 300 mM, 5-HT stimulated [(35)S]GTPgammaS binding, basal Galpha(i3) activation was low and methiothepin was inactive. In contrast, at 10 mM NaCl, basal activity was enhanced and the inverse agonist activity of methiothepin was accentuated. Under these conditions, 5-HT decreased Galpha(i3) activation. 5. In conclusion, at h5-HT(1B) receptors expressed in CHO cells: (i) inverse agonist induced inhibition of Galpha(i3), and its reversal by S18127, reveals constitutive activation of this Galpha subunit; (ii) constitutive Galpha(i3) activation can be quantified by isotopic dilution [(35)S]GTPgammaS binding and (iii) decreasing NaCl concentrations enhances Galpha(i3

  18. Absolute nuclear material assay

    Science.gov (United States)

    Prasad, Manoj K [Pleasanton, CA; Snyderman, Neal J [Berkeley, CA; Rowland, Mark S [Alamo, CA

    2010-07-13

    A method of absolute nuclear material assay of an unknown source comprising counting neutrons from the unknown source and providing an absolute nuclear material assay utilizing a model to optimally compare to the measured count distributions. In one embodiment, the step of providing an absolute nuclear material assay comprises utilizing a random sampling of analytically computed fission chain distributions to generate a continuous time-evolving sequence of event-counts by spreading the fission chain distribution in time.

  19. Fuel cell end plate structure

    Science.gov (United States)

    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. [Conventional plate osteosynthesis].

    Science.gov (United States)

    Klaue, K

    2010-02-01

    Consolidation of bone is an essential clinical problem when treating fractures, fixing osteotomies and fusing joints. In most cases, the means of fixation are plates and screws. The goal is functional postoperative therapy by moving the adjacent joints and thus avoiding the deleterious disadvantages of long-lasting articular immobilization. Pre-operative planning, surgical approach, a good understanding of the precise mechanics of the structure and the biological answer for the various tissues are prerequisites of successful osteosynthesis. The choice of implants and the application of their versatility, as well as their adaptation to individual cases are the key to good results.

  1. Plate Full of Color

    Centers for Disease Control (CDC) Podcasts

    2008-08-04

    The Eagle Books are a series of four books that are brought to life by wise animal characters - Mr. Eagle, Miss Rabbit, and Coyote - who engage Rain That Dances and his young friends in the joy of physical activity, eating healthy foods, and learning from their elders about health and diabetes prevention. Plate Full of Color teaches the value of eating a variety of colorful and healthy foods.  Created: 8/4/2008 by National Center for Chronic Disease Prevention and Health Promotion (NCCDPHP).   Date Released: 8/5/2008.

  2. Endogenous Locus Reporter Assays.

    Science.gov (United States)

    Liu, Yaping; Hermes, Jeffrey; Li, Jing; Tudor, Matthew

    2018-01-01

    Reporter gene assays are widely used in high-throughput screening (HTS) to identify compounds that modulate gene expression. Traditionally a reporter gene assay is built by cloning an endogenous promoter sequence or synthetic response elements in the regulatory region of a reporter gene to monitor transcriptional activity of a specific biological process (exogenous reporter assay). In contrast, an endogenous locus reporter has a reporter gene inserted in the endogenous gene locus that allows the reporter gene to be expressed under the control of the same regulatory elements as the endogenous gene, thus more accurately reflecting the changes seen in the regulation of the actual gene. In this chapter, we introduce some of the considerations behind building a reporter gene assay for high-throughput compound screening and describe the methods we have utilized to establish 1536-well format endogenous locus reporter and exogenous reporter assays for the screening of compounds that modulate Myc pathway activity.

  3. Plating on difficult-to-plate metals: what's new

    International Nuclear Information System (INIS)

    Wiesner, H.J.

    1980-01-01

    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

  4. An Automated Method for High-Throughput Screening of Arabidopsis Rosette Growth in Multi-Well Plates and Its Validation in Stress Conditions

    Czech Academy of Sciences Publication Activity Database

    De Diego, N.; Fürst, T.; Humplík, Jan; Ugena, L.; Podlešáková, K.; Spíchal, L.

    2017-01-01

    Roč. 8, OCT 4 (2017), č. článku 1702. ISSN 1664-462X R&D Projects: GA MŠk(CZ) LO1204 Institutional support: RVO:61389030 Keywords : salt stress * chlorophyll fluorescence * salinity tolerance * plant-responses * cold-tolerance * water-deficit * thaliana * selection * platform * reveals * high-throughput screening assay * Arabidopsis * multi-well plates * rosette growth * stress conditions Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 4.298, year: 2016

  5. A computed microtomography method for understanding epiphyseal growth plate fusion

    Science.gov (United States)

    Staines, Katherine A.; Madi, Kamel; Javaheri, Behzad; Lee, Peter D.; Pitsillides, Andrew A.

    2017-12-01

    The epiphyseal growth plate is a developmental region responsible for linear bone growth, in which chondrocytes undertake a tightly regulated series of biological processes. Concomitant with the cessation of growth and sexual maturation, the human growth plate undergoes progressive narrowing, and ultimately disappears. Despite the crucial role of this growth plate fusion ‘bridging’ event, the precise mechanisms by which it is governed are complex and yet to be established. Progress is likely hindered by the current methods for growth plate visualisation; these are invasive and largely rely on histological procedures. Here we describe our non-invasive method utilising synchrotron x-ray computed microtomography for the examination of growth plate bridging, which ultimately leads to its closure coincident with termination of further longitudinal bone growth. We then apply this method to a dataset obtained from a benchtop microcomputed tomography scanner to highlight its potential for wide usage. Furthermore, we conduct finite element modelling at the micron-scale to reveal the effects of growth plate bridging on local tissue mechanics. Employment of these 3D analyses of growth plate bone bridging is likely to advance our understanding of the physiological mechanisms that control growth plate fusion.

  6. Investigation on filter method for smoothing spiral phase plate

    Science.gov (United States)

    Zhang, Yuanhang; Wen, Shenglin; Luo, Zijian; Tang, Caixue; Yan, Hao; Yang, Chunlin; Liu, Mincai; Zhang, Qinghua; Wang, Jian

    2018-03-01

    Spiral phase plate (SPP) for generating vortex hollow beams has high efficiency in various applications. However, it is difficult to obtain an ideal spiral phase plate because of its continuous-varying helical phase and discontinued phase step. This paper describes the demonstration of continuous spiral phase plate using filter methods. The numerical simulations indicate that different filter method including spatial domain filter, frequency domain filter has unique impact on surface topography of SPP and optical vortex characteristics. The experimental results reveal that the spatial Gaussian filter method for smoothing SPP is suitable for Computer Controlled Optical Surfacing (CCOS) technique and obtains good optical properties.

  7. Multidimensional Normalization to Minimize Plate Effects of Suspension Bead Array Data.

    Science.gov (United States)

    Hong, Mun-Gwan; Lee, Woojoo; Nilsson, Peter; Pawitan, Yudi; Schwenk, Jochen M

    2016-10-07

    Enhanced by the growing number of biobanks, biomarker studies can now be performed with reasonable statistical power by using large sets of samples. Antibody-based proteomics by means of suspension bead arrays offers one attractive approach to analyze serum, plasma, or CSF samples for such studies in microtiter plates. To expand measurements beyond single batches, with either 96 or 384 samples per plate, suitable normalization methods are required to minimize the variation between plates. Here we propose two normalization approaches utilizing MA coordinates. The multidimensional MA (multi-MA) and MA-loess both consider all samples of a microtiter plate per suspension bead array assay and thus do not require any external reference samples. We demonstrate the performance of the two MA normalization methods with data obtained from the analysis of 384 samples including both serum and plasma. Samples were randomized across 96-well sample plates, processed, and analyzed in assay plates, respectively. Using principal component analysis (PCA), we could show that plate-wise clusters found in the first two components were eliminated by multi-MA normalization as compared with other normalization methods. Furthermore, we studied the correlation profiles between random pairs of antibodies and found that both MA normalization methods substantially reduced the inflated correlation introduced by plate effects. Normalization approaches using multi-MA and MA-loess minimized batch effects arising from the analysis of several assay plates with antibody suspension bead arrays. In a simulated biomarker study, multi-MA restored associations lost due to plate effects. Our normalization approaches, which are available as R package MDimNormn, could also be useful in studies using other types of high-throughput assay data.

  8. Plate Tearing by a Cone

    DEFF Research Database (Denmark)

    Simonsen, Bo Cerup

    1998-01-01

    The present paper is concerned with steady-state plate tearing by a cone. This is a scenario where a cone is forced through a ductile metal plate with a constant lateral tip penetration in a motion in the plane of the plate. The considered process could be an idealisation of the damage, which...... as for the out-of-plane reaction force. (C) 1998 Elsevier Science Ltd. All rights reserved....

  9. Bipolar Plates for PEM Systems

    OpenAIRE

    Lædre, Sigrid

    2016-01-01

    Summary of thesis: The Bipolar Plate (BPP) is an important component in both Proton Exchange Membrane Fuel Cells (PEMFCs) and Proton Exchange Membrane Water Electrolyzers (PEMWEs). Bipolar plate material and processing constitutes for a large fraction of the cost and weight of a PEM cell stack. The main tasks for the bipolar plates in both systems are to separate single cell in a stack, conduct current between single cells and remove heat from active areas. In addition, the BPPs distribu...

  10. Scintillator plate calorimetry

    International Nuclear Information System (INIS)

    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

  11. Reviewing metallic PEMFC bipolar plates

    Energy Technology Data Exchange (ETDEWEB)

    Wang, H.; Turner, J.A. [National Renewable Energy Laboratory, Golden, CO (United States)

    2010-08-15

    A bipolar plate is one of the most important components in a polymer exchange membrane fuel cell (PEMFC) stack and has multiple functions. Metallic bipolar plate candidates have advantages over composite rivals in excellent electrical and thermal conductivity, good mechanical strength, high chemical stability, very wide alloy choices, low cost and, most importantly, existing pathways for high-volume, high-speed mass production. The challenges with metallic bipolar plates are the higher contact resistance and possible corrosion products, which may contaminate the membrane electrode assembly. This review evaluates the candidate metallic and coating materials for bipolar plates and gives the perspective of the research trends. (Abstract Copyright [2010], Wiley Periodicals, Inc.)

  12. Solid phase assays

    International Nuclear Information System (INIS)

    Reese, M.G.; Johnson, L.R.; Ransom, D.K.

    1980-01-01

    In a solid phase assay for quantitative determination of biological and other analytes, a sample such as serum is contacted with a receptor for the analyte being assayed, the receptor being supported on a solid support. No tracer for the analyte is added to the sample before contacting with the receptor; instead the tracer is contacted with the receptor after unbound analyte has been removed from the receptor. The assay can be otherwise performed in a conventional manner but can give greater sensitivity. (author)

  13. New low-viscosity overlay medium for viral plaque assays

    Directory of Open Access Journals (Sweden)

    Garten Wolfgang

    2006-08-01

    Full Text Available Abstract Background Plaque assays in cell culture monolayers under solid or semisolid overlay media are commonly used for quantification of viruses and antiviral substances. To overcome the pitfalls of known overlays, we tested suspensions of microcrystalline cellulose Avicel RC/CL™ as overlay media in the plaque and plaque-inhibition assay of influenza viruses. Results Significantly larger plaques were formed under Avicel-containing media, as compared to agar and methylcellulose (MC overlay media. The plaque size increased with decreasing Avicel concentration, but even very diluted Avicel overlays (0.3% ensured formation of localized plaques. Due to their low viscosity, Avicel overlays were easier to use than methylcellulose overlays, especially in the 96-well culture plates. Furthermore, Avicel overlay could be applied without prior removal of the virus inoculum thus facilitating the assay and reducing chances of cross-contamination. Using neuraminidase inhibitor oseltamivir carboxylate, we demonstrated applicability of the Avicel-based plaque reduction assay for testing of antiviral substances. Conclusion Plaque assay under Avicel-containing overlay media is easier, faster and more sensitive than assays under agar- and methylcellulose overlays. The assay can be readily performed in a 96-well plate format and seems particularly suitable for high-throughput virus titrations, serological studies and experiments on viral drug sensitivity. It may also facilitate work with highly pathogenic agents performed under hampered conditions of bio-safety labs.

  14. A quantitative comet infection assay for influenza virus

    Science.gov (United States)

    Lindsay, Stephen M.; Timm, Andrea; Yin, John

    2011-01-01

    Summary The virus comet assay is a cell-based virulence assay used to evaluate an antiviral drug or antibody against a target virus. The comet assay differs from the plaque assay in allowing spontaneous flows in 6-well plates to spread virus. When implemented quantitatively the comet assay has been shown to have an order-of-magnitude greater sensitivity to antivirals than the plaque assay. In this study, a quantitative comet assay for influenza virus is demonstrated, and is shown to have a 13-fold increase in sensitivity to ribavirin. AX4 cells (MDCK cells with increased surface concentration of α2–6 sialic acid, the influenza virus receptor) have reduced the comet size variability relative to MDCK cells, making them a better host cell for use in this assay. Because of enhanced antiviral sensitivity in flow-based assays, less drug is required, which could lead to lower reagent costs, reduced cytotoxicity, and fewer false-negative drug screen results. The comet assay also serves as a readout of flow conditions in the well. Observations from comets formed at varying humidity levels indicate a role for evaporation in the mechanism of spontaneous fluid flow in wells. PMID:22155578

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

    Science.gov (United States)

    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. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. A Quantitative Fluorescence-Based Lipase Assay

    Directory of Open Access Journals (Sweden)

    Giovanna Lomolino

    2012-01-01

    Full Text Available An easy and fast gel diffusion assay for detecting and monitoring lipase activity by quantification of fluorescein is described. By measuring the intensity of fluorescein, it is possible to obtain a calibration curve with a regression coefficient better than by using the radius of fluorescent haloes. Through the quantification of fluorescence intensity of fluorescein released after the hydrolysis of a fluorescent ester, fluorescein dibutyrate, used as substrate in agar plates, commercial and skimmed milk lipase activity were studied. Moreover, with this method, lipase activity can be monitored in reaction medium that contains compounds which are affected by turbidity or cause measurement interference for UV-spectrophotometer and fluorimeter. In this experiment, boiled skimmed milk was dispersed in the agar gel with fluorescein dibutyrate, and it was used as a reaction medium to mimic natural conditions. The development of such an assay has a potential for applications in industries ranging from pharmaceuticals to food production and monitoring.

  17. Factor IX assay

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003679.htm Factor IX assay To use the sharing features on ... M. is also a founding member of Hi-Ethics and subscribes to the principles of the Health ...

  18. Factor VIII assay

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003678.htm Factor VIII assay To use the sharing features on ... M. is also a founding member of Hi-Ethics and subscribes to the principles of the Health ...

  19. Factor II assay

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003674.htm Factor II assay To use the sharing features on ... M. is also a founding member of Hi-Ethics and subscribes to the principles of the Health ...

  20. Factor VII assay

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003676.htm Factor VII assay To use the sharing features on ... M. is also a founding member of Hi-Ethics and subscribes to the principles of the Health ...

  1. Microbead agglutination based assays

    KAUST Repository

    Kodzius, Rimantas; Castro, David; Foulds, Ian G.; Parameswaran, Ash M.; Sumanpreet, K. Chhina

    2013-01-01

    We report a simple and rapid room temperature assay for point-of-care (POC) testing that is based on specific agglutination. Agglutination tests are based on aggregation of microbeads in the presence of a specific analyte thus enabling

  2. Modeling particulate removal in plate-plate and wire-plate electrostatic precipitators

    Directory of Open Access Journals (Sweden)

    S Ramechecandane

    2016-09-01

    Full Text Available The present study is concerned with the modeling of electrically charged particles in a model plate-plate and a single wire-plate electrostatic precipitator (ESP. The particle concentration distributions for both a plate-plate and a wire-plate ESP are calculated using a modified drift flux model. Numerical investigations are performed using the modified drift flux model for particle number concentration, in addition to the RNG k - ε model for the mean turbulent flow field and the Poisson equation for the electric field. The proposed model and the outlined methodology for coupling the flow field, electric field, charging kinetics and particle concentration is applied to two model precipitators that are truly representative of a wide class of commercialized ESPs. The present investigation is quite different from the earlier studies as it does not make assumptions like a homogeneous electric field or an infinite turbulent diffusivity. The electric field calculated is a strong function of position and controls the migration velocity of particles. Hence, the proposed model can be implemented in a flow solver to obtain a full-fledged solution for any kind of ESP with no limitations on the particle number concentration, as encountered in a Lagrangian approach. The effect of turbulent diffusivity on particle number concentration in a plate-plate ESP is investigated in detail and the results obtained are compared with available experimental data. Similarly, the effect of particle size/diameter and applied electric potential on the accumulative collection performance in the case of a wire-plate ESP is studied and the results obtained are compared with available numerical data. The numerical results obtained using the modified drift flux model for both the plate-plate and wire-plate ESP are in close agreement with available experimental and numerical data.

  3. Laterally Loaded Nail-Plates

    DEFF Research Database (Denmark)

    Nielsen, Jacob; Rathkjen, Arne

    Load-displacement curves from about 200 short-term and laterally loaded nail-plate joints are analysed. The nail-plates are from Gang-Nail Systems, type GNA 20 S. The test specimens and the measuring systems are described. The tests are divided into 32 different series. The influence of the number...

  4. MyPlate Food Guide

    Science.gov (United States)

    ... Safe Videos for Educators Search English Español MyPlate Food Guide KidsHealth / For Teens / MyPlate Food Guide What's ... and other sugary drinks. Avoid large portions . Five Food Groups Different food groups have different nutrients and ...

  5. Scintillating plate calorimeter optical design

    International Nuclear Information System (INIS)

    McNeil, R.; Fazely, A.; Gunasingha, R.; Imlay, R.; Lim, J.

    1990-01-01

    A major technical challenge facing the builder of a general purpose detector for the SSC is to achieve an optimum design for the calorimeter. Because of its fast response and good energy resolution, scintillating plate sampling calorimeters should be considered as a possible technology option. The work of the Scintillating Plate Calorimeter Collaboration is focused on compensating plate calorimeters. Based on experimental and simulation studies, it is expected that a sampling calorimeter with alternating layers of high-Z absorber (Pb, W, DU, etc.) and plastic scintillator can be made compensating (e/h = 1.00) by suitable choice of the ratio of absorber/scintillator thickness. Two conceptual designs have been pursued by this subsystem collaboration. One is based on lead as the absorber, with read/out of the scintillator plates via wavelength shifter fibers. The other design is based on depleted uranium as the absorber with wavelength shifter (WLS) plate readout. Progress on designs for the optical readout of a compensating scintillator plate calorimeter are presented. These designs include readout of the scintillator plates via wavelength shifter plates or fiber readout. Results from radiation damage studies of the optical components are presented

  6. Fundamental processes in ion plating

    International Nuclear Information System (INIS)

    Mattox, D.M.

    1980-01-01

    Ion plating is a generic term applied to film deposition processes in which the substrate surface and/or the depositing film is subjected to a flux of high energy particles sufficient to cause changes in the interfacial region of film properties compared to a nonbombarded deposition. Ion plating is being accepted as an alternative coating technique to sputter deposition, vacuum evaporation and electroplating. In order to intelligently choose between the various deposition techniques, the fundamental mechanisms, relating to ion plating, must be understood. This paper reviews the effects of low energy ion bombardment on surfaces, interface formation and film development as they apply to ion plating and the implementation and applications of the ion plating process

  7. A simple colorimetric assay for detection of amplified Mycobacterium leprae DNA

    NARCIS (Netherlands)

    van der Vliet, G. M.; de Wit, M. Y.; Klatser, P. R.

    1993-01-01

    A colorimetric assay for the detection of PCR-products is described. The assay is based on amplification of DNA in the presence of digoxigenin-dUTP. After immobilization of the PCR products to a microtitre plate, amplified DNA could be detected colorimetrically. The sensitivity of this colorimetric

  8. Detection of Microbial Growth on Polycyclic Aromatic Hydrocarbons in Microtiter Plates by Using the Respiration Indicator WST-1

    OpenAIRE

    Johnsen, Anders R.; Bendixen, Karen; Karlson, Ulrich

    2002-01-01

    We have developed a microtiter plate method for screening a large number of bacterial isolates for the ability to grow on different crystalline polycyclic aromatic hydrocarbons (PAHs). Growth on PAHs cannot easily be determined with standard growth assays because of the very low aqueous solubility and bioavailability of the PAHs. Our microtiter plate assay utilizes a new water-soluble respiration indicator, WST-1 {4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate}...

  9. Isolated oestrogen deficiency in male 30-year-old: persistent growth plates with severe osteopenia

    International Nuclear Information System (INIS)

    Smith, T.; Roberts, J.; Howman-Giles, R.; Cowell, C.; Jeremy, R.

    2000-01-01

    Full text: A 31-year-old male presented with right rib pain and generalised skeletal symptoms. He has a past history of multiple fractures following trauma. No history of childhood fractures. Asthma (No steroids). A product of a consanguineous marriage, he has one child aged 7. Examination showed 178.6cm male with normal sexual characteristics. No abnormality detected apart from tenderness over right ribs. Bone scan showed active growth plates and right and left rib fractures. X-rays demonstrated a bone age of 15 1/2 - 16 yrs and a compression fracture of L2. His bone mineral density is severely reduced. Metabolic investigations revealed Testosterone 27.4 nmol (N 11-35), oestradiol < 70 pmol/L, ultrasensitive assay 14 and 17 pmol/L (consistent with 8-year-old male), LH N, FSH N, 46 XY Karyotype. Alkaline phosphatase. 148 (N<120), Normal glucose tolerance test. This patient illustrates a very rare condition of oestrogen deficiency in a male, probably due to aromatase deficiency. This enzyme converts testosterone to oestradiol. It illustrates the role of oestrogen in fusing growth plates and maintaining bone mass in males with otherwise normal androgen levels. A similar clinical picture can result from an oestrogen receptor abnormality. Copyright (2000) The Australian and New Zealand Society of Nuclear Medicine Inc

  10. The Golosyiv plate archive digitisation

    Science.gov (United States)

    Sergeeva, T. P.; Sergeev, A. V.; Pakuliak, L. K.; Yatsenko, A. I.

    2007-08-01

    The plate archive of the Main Astronomical Observatory of the National Academy of Sciences of Ukraine (Golosyiv, Kyiv) includes about 85 000 plates which have been taken in various observational projects during 1950-2005. Among them are about 25 000 of direct northern sky area plates and more than 600 000 plates containing stellar, planetary and active solar formations spectra. Direct plates have a limiting magnitude of 14.0-16.0 mag. Since 2002 we have been organising the storage, safeguarding, cataloguing and digitization of the plate archive. The very initial task was to create the automated system for detection of astronomical objects and phenomena, search of optical counterparts in the directions of gamma-ray bursts, research of long period, flare and other variable stars, search and rediscovery of asteroids, comets and other Solar System bodies to improve the elements of their orbits, informational support of CCD observations and space projects, etc. To provide higher efficiency of this work we have prepared computer readable catalogues and database for 250 000 direct wide field plates. Now the catalogues have been adapted to Wide Field Plate Database (WFPDB) format and integrated into this world database. The next step will be adaptation of our catalogues, database and images to standards of the IVOA. Some magnitude and positional accuracy estimations for Golosyiv archive plates have been done. The photometric characteristics of the images of NGC 6913 cluster stars on two plates of the Golosyiv's double wide angle astrograph have been determined. Very good conformity of the photometric characteristics obtained with external accuracies of 0.13 and 0.15 mag. has been found. The investigation of positional accuracy have been made with A3± format fixed bed scanner (Microtek ScanMaker 9800XL TMA). It shows that the scanner has non-detectable systematic errors on the X-axis, and errors of ± 15 μm on the Y-axis. The final positional errors are about ± 2 μm (

  11. Indonesian Landforms and Plate Tectonics

    Directory of Open Access Journals (Sweden)

    Herman Th. Verstappen

    2014-06-01

    Full Text Available DOI: 10.17014/ijog.v5i3.103The horizontal configuration and vertical dimension of the landforms occurring in the tectonically unstable parts of Indonesia were resulted in the first place from plate tectonics. Most of them date from the Quaternary and endogenous forces are ongoing. Three major plates – the northward moving Indo-Australian Plate, the south-eastward moving SE-Asian Plate and the westward moving Pacific Plate - meet at a plate triple-junction situated in the south of New Guinea’s Bird’s Head. The narrow North-Moluccan plate is interposed between the Asia and Pacific. It tapers out northward in the Philippine Mobile Belt and is gradually disappearing. The greatest relief amplitudes occur near the plate boundaries: deep ocean trenches are associated with subduction zones and mountain ranges with collision belts. The landforms of the more stable areas of the plates date back to a more remote past and, where emerged, have a more subdued relief that is in the first place related to the resistance of the rocks to humid tropical weathering Rising mountain ranges and emerging island arcs are subjected to rapid humid-tropical river erosions and mass movements. The erosion products accumulate in adjacent sedimentary basins where their increasing weight causes subsidence by gravity and isostatic compensations. Living and raised coral reefs, volcanoes, and fault scarps are important geomorphic indicators of active plate tectonics. Compartmental faults may strongly affect island arcs stretching perpendicular to the plate movement. This is the case on Java. Transcurrent faults and related pull-apart basins are a leading factor where plates meet at an angle, such as on Sumatra. The most complicated situation exists near the triple-junction and in the Moluccas. Modern research methods, such as GPS measurements of plate movements and absolute dating of volcanic outbursts and raised coral reefs are important tools. The mega-landforms resulting

  12. Assay method and compositions

    International Nuclear Information System (INIS)

    1977-01-01

    Methods are described for measuring catecholamine levels in human and animal body fluids and tissues using the catechol-O-methyl-transferase (COMT) radioassay. The assay involves incubating the biological sample with COMT and the tritiated methyl donor, S-adenosyl-L-methionine( 3 H)-methyl. The O-methylated ( 3 H) epinephrine and/or norepinephrine are extracted and oxidised to vanillin- 3 H which in turn is extracted and its radioactivity counted. When analysing dopamine levels the assay is extended by vanillin- 3 H and raising the pH of the aqueous periodate phase from which O-methylated ( 3 H) dopamine is extracted and counted. The assay may be modified depending on whether measurements of undifferentiated total endogenous catecholamine levels or differential analyses of the catecholamine levels are being performed. The sensitivity of the assay can be as low as 5 picograms for norepinephrine and epinephrine and 12 picograms for dopamine. The assemblance of the essential components of the assay into a kit for use in laboratories is also described. (U.K.)

  13. Splitting in Dual-Phase 590 high strength steel plates

    International Nuclear Information System (INIS)

    Yang Min; Chao, Yuh J.; Li Xiaodong; Tan Jinzhu

    2008-01-01

    Charpy V-notch impact tests on 5.5 mm thick, hot-rolled Dual-Phase 590 (DP590) steel plate were evaluated at temperatures ranging from 90 deg. C to -120 deg. C. Similar tests on 2.0 mm thick DP590 HDGI steel plate were also conducted at room temperature. Splitting or secondary cracks was observed on the fractured surfaces. The mechanisms of the splitting were then investigated. Fracture surfaces were analyzed by optical microscope (OM) and scanning electron microscope (SEM). Composition of the steel plates was determined by electron probe microanalysis (EPMA). Micro Vickers hardness of the steel plates was also surveyed. Results show that splitting occurred on the main fractured surfaces of hot-rolled steel specimens at various testing temperatures. At temperatures above the ductile-brittle-transition-temperature (DBTT), -95 deg. C, where the fracture is predominantly ductile, the length and amount of splitting decreased with increasing temperature. At temperatures lower than the DBTT, where the fracture is predominantly brittle, both the length and width of the splitting are insignificant. Splitting in HDGI steel plates only appeared in specimens of T-L direction. The analysis revealed that splitting in hot-rolled plate is caused by silicate and carbide inclusions while splitting in HDGI plate results from strip microstructure due to its high content of manganese and low content of silicon. The micro Vickers hardness of either the inclusions or the strip microstructures is higher than that of the respective base steel

  14. Plasminogen activator activity and plasma-coagulum lysis measured by use of optimized fibrin gel structure preformed in microtiter plates

    DEFF Research Database (Denmark)

    Sidelmann, Johannes Jakobsen; Jespersen, J; Gram, J

    1995-01-01

    We introduce a new fibrin plate assay performed in microtiter plates. By means of spectroscopic studies we optimized the structure of the fibrin gel and then used the optimized fibrin gel to determine plasminogen activator activity. Plasminogen activator solutions were applied on top of the fibri...

  15. Rover waste assay system

    Energy Technology Data Exchange (ETDEWEB)

    Akers, D.W.; Stoots, C.M.; Kraft, N.C.; Marts, D.J. [Idaho National Engineering Lab., Idaho Falls, ID (United States)

    1997-11-01

    The Rover Waste Assay System (RWAS) is a nondestructive assay system designed for the rapid assay of highly-enriched {sup 235}U contaminated piping, tank sections, and debris from the Rover nuclear rocket fuel processing facility at the Idaho Chemical Processing Plant. A scanning system translates a NaI(Tl) detector/collimator system over the structural components where both relative and calibrated measurements for {sup 137}Cs are made. Uranium-235 concentrations are in operation and is sufficiently automated that most functions are performed by the computer system. These functions include system calibration, problem identification, collimator control, data analysis, and reporting. Calibration of the system was done through a combination of measurements on calibration standards and benchmarked modeling. A description of the system is presented along with the methods and uncertainties associated with the calibration and analysis of the system for components from the Rover facility. 4 refs., 2 figs., 4 tabs.

  16. Rover waste assay system

    International Nuclear Information System (INIS)

    Akers, D.W.; Stoots, C.M.; Kraft, N.C.; Marts, D.J.

    1997-01-01

    The Rover Waste Assay System (RWAS) is a nondestructive assay system designed for the rapid assay of highly-enriched 235 U contaminated piping, tank sections, and debris from the Rover nuclear rocket fuel processing facility at the Idaho Chemical Processing Plant. A scanning system translates a NaI(Tl) detector/collimator system over the structural components where both relative and calibrated measurements for 137 Cs are made. Uranium-235 concentrations are in operation and is sufficiently automated that most functions are performed by the computer system. These functions include system calibration, problem identification, collimator control, data analysis, and reporting. Calibration of the system was done through a combination of measurements on calibration standards and benchmarked modeling. A description of the system is presented along with the methods and uncertainties associated with the calibration and analysis of the system for components from the Rover facility. 4 refs., 2 figs., 4 tabs

  17. Radioreceptor assay for insulin

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, Kazuo [Tokyo Univ. (Japan). Faculty of Medicine

    1975-04-01

    Radioreceptor assay of insulin was discussed from the aspects of the measuring method, its merits and problems to be solved, and its clinical application. Rat liver 10 x g pellet was used as receptor site, and enzymatic degradation of insulin by the system contained in this fraction was inhibited by adding 1 mM p-CMB. /sup 125/I-labelled porcine insulin was made by lactoperoxidase method under overnight incubation at 4/sup 0/C and later purification by Sephadex G-25 column and Whatman CF-11 cellulose powder. Dog pancreatic vein serum insulin during and after the glucose load was determined by radioreceptor assay and radioimmunoassay resulting that both measurements accorded considerably. Radioreceptor assay would clarify the pathology of disorders of glucose metabolism including diabetes.

  18. [Tibiotalocalcaneal arthrodesis using a humeral locking plate].

    Science.gov (United States)

    Cabrera Méndez, M P; Gamba, C; Hernández, E; Molano, J; Andrade, J C

    2016-01-01

    To describe the results of tibiotalocalcáneal arthrodesis (TTC) using a humeral locking plate. A retrospective, observational study was conducted between January 2007 and December 2013 in the Hospital Militar Central de Bogotá. The study included patients with symptomatic osteoarthritis diagnosed clinically and radiologically, and who underwent TTC arthrodesis using a humeral locking plate with a minimum follow up of 6 months. The total number patients was 35, of whom 7 (20%) were women and 28 (80%) men, with a mean age 36.3 years (19.77). 74% with post-traumatic arthritis, most of them secondary to gunshot wounds and fragmentation weapons, and neuropathic in 20%. An autogenous graft was used in 13 cases, and 14 cases using both, with a mean consolidation time of 4.37 months. Complications include, delayed union in 3 cases, and surgical site infection in 4. The postoperative functionality (AOFAS) mean was 66.7/100 points, with a score of 2.35 on a visual analogue pain scale. TTC arthrodesis using a humeral locking plate is a suitable option for fixing this type of arthrodesis, with a low rate of complications, and postoperative results that revealed satisfactory improvement in pain and consolidation. Copyright © 2015 SECOT. Published by Elsevier Espana. All rights reserved.

  19. High-throughput screening of carbohydrate-degrading enzymes using novel insoluble chromogenic substrate assay kits

    DEFF Research Database (Denmark)

    Schückel, Julia; Kracun, Stjepan Kresimir; Willats, William George Tycho

    2016-01-01

    for this is that advances in genome and transcriptome sequencing, together with associated bioinformatics tools allow for rapid identification of candidate CAZymes, but technology for determining an enzyme's biochemical characteristics has advanced more slowly. To address this technology gap, a novel high-throughput assay...... CPH and ICB substrates are provided in a 96-well high-throughput assay system. The CPH substrates can be made in four different colors, enabling them to be mixed together and thus increasing assay throughput. The protocol describes a 96-well plate assay and illustrates how this assay can be used...... for screening the activities of enzymes, enzyme cocktails, and broths....

  20. Scintillation proximity assay

    International Nuclear Information System (INIS)

    Hart, H.

    1980-01-01

    In a method of immunological assay two different classes of particles which interact at short distances to produce characteristic detectable signals are employed in a modification of the usual latex fixation test. In one embodiment an aqueous suspension of antigen coated tritiated latex particles (LH) and antigen coated polystyrene scintillant particles (L*) is employed to assay antibody in the aqueous medium. The amount of (LH) (L*) dimer formation and higher order aggregation induced and therefore the concentration of antibody (or antigen) present which caused the aggregation can be determined by using standard liquid scintillation counting equipment. (author)

  1. Assays for calcitonin receptors

    International Nuclear Information System (INIS)

    Teitelbaum, A.P.; Nissenson, R.A.; Arnaud, C.D.

    1985-01-01

    The assays for calcitonin receptors described focus on their use in the study of the well-established target organs for calcitonin, bone and kidney. The radioligand used in virtually all calcitonin binding studies is 125 I-labelled salmon calcitonin. The lack of methionine residues in this peptide permits the use of chloramine-T for the iodination reaction. Binding assays are described for intact bone, skeletal plasma membranes, renal plasma membranes, and primary kidney cell cultures of rats. Studies on calcitonin metabolism in laboratory animals and regulation of calcitonin receptors are reviewed

  2. Detection and removal of spatial bias in multiwell assays.

    Science.gov (United States)

    Lachmann, Alexander; Giorgi, Federico M; Alvarez, Mariano J; Califano, Andrea

    2016-07-01

    Multiplex readout assays are now increasingly being performed using microfluidic automation in multiwell format. For instance, the Library of Integrated Network-based Cellular Signatures (LINCS) has produced gene expression measurements for tens of thousands of distinct cell perturbations using a 384-well plate format. This dataset is by far the largest 384-well gene expression measurement assay ever performed. We investigated the gene expression profiles of a million samples from the LINCS dataset and found that the vast majority (96%) of the tested plates were affected by a significant 2D spatial bias. Using a novel algorithm combining spatial autocorrelation detection and principal component analysis, we could remove most of the spatial bias from the LINCS dataset and show in parallel a dramatic improvement of similarity between biological replicates assayed in different plates. The proposed methodology is fully general and can be applied to any highly multiplexed assay performed in multiwell format. ac2248@columbia.edu Supplementary data are available at Bioinformatics online. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  3. Plate shell structures of glass

    DEFF Research Database (Denmark)

    Bagger, Anne

    to their curved shape. A plate shell structure maintains a high stiffness-to-weight ratio, while facilitating the use of plane structural elements. The study focuses on using laminated glass panes for the load bearing facets. Various methods of generating a plate shell geometry are suggested. Together with Ghent......, such as facet size, imperfections, and connection characteristics. The critical load is compared to that of a similar, but smoothly curved, shell structure. Based on the investigations throughout the study, a set of guidelines for the structural design of plate shells of glass is proposed....

  4. Plating on stainless steel alloys

    International Nuclear Information System (INIS)

    Dini, J.W.; Johnson, H.R.

    1981-01-01

    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

  5. A novel microtiter plate radioimmunoassay of insulin autoantibody

    International Nuclear Information System (INIS)

    Huang Gan; Li Zhangwei; Jin Helai; Wang Xia; Wang Jianping; Zhou Zhiguang

    2009-01-01

    Objective: Insulin autoantibody (IAA) is known to exist in sera of type 1 diabetes mellitus (T1DM) patients and pre-T1DM individuals. The aim of this study was to establish a novel mierotiter plate radioimmunoassay (RIA) for IAA and evaluate its clinical value. Methods: Diluted 125 I-insulin was mixed with 5 μl serum samples in a 96-well microtiter plate and then incubated for 72 h on an orbital plate shaker (4 degree C). The immunocomplexes were transferred to another protein a coated Millipore plate, and then the plate was washed with Tri-Buffered Saline Tween-20 (TBT) buffer. Counts per minute (CPM) was measured with liquid scintillation and luminescence counter. The positive cut-off point of IAA index was defined as ≥0.06 based on the 99-percentile of the distribution in 317 healthy individuals. The specificity and sensitivity of the assay were calculated from the samples provided by the fourth Diabetes Autoantibodies Standardization Program (DASP 2005). The IAA levels were determined in 71 T1DM and 551 newly diagnosed type 2 diabetes (T2DM) patients, and 317 healthy controls. The t test, non-parametric test, χ 2 test and linear correlation analysis were performed on the data using SPSS 11.5 software. The concordance rate was estimated with Kappa value. Results: (1) The optimized testing condition was described as 2 x 10 4 CPM of 125 I-insulin, 5 μl serum sample and slowly horizontal shaking for 72 h. (2) The intra-assay CV was 4.8%-8.9% and inter-assay CV was 6.4%-10.5%. Based on DASP 2005 samples, the specificity and sensitivity of the assay were 97% (97/100) and 50% (25/50), respectively. Ninety-six serum samples with different IAA levels were selected and tested to compare between our new method and a domestic IAA RIA kit. The results showed that the IAA indices from the two methods were positively correlated (r= 0.678, P<0.001). The concordance rate was 72.9% (Kappa value=0.402). There were 25 samples with discordant results, which were positive for

  6. Cytotoxicity difference of 316L stainless steel and titanium reconstruction plate

    Directory of Open Access Journals (Sweden)

    Ni Putu Mira Sumarta

    2011-03-01

    Full Text Available Background: Pure titanium is the most biocompatible material today and used as a gold standard for metallic implants. However, stainless steel is still being used as implants because of its strength, ductility, lower price, corrosion resistant and biocompatibility. Purpose: This study was done to revealed the cytotoxicity difference between reconstruction plate made of 316L stainless steel and of commercially pure (CP titanium in baby hamster kidney-21 (BHK-21 fibroblast culture through MTT assay. Methods: Eight samples were prepared from reconstruction plates made of stainless steel type 316L grade 2 (Coen’s reconstruction plate® that had been cut into cylindrical form of 2 mm in diameter and 3 mm long. The other one were made of CP titanium (STEMA Gmbh® of 2 mm in diameter and 2,2 mm long; and had been cleaned with silica paper and ultrasonic cleaner, and sterilized in autoclave at 121° C for 20 minutes.9 Both samples were bathed into microplate well containing 50 μl of fibroblast cells with 2 x 105 density in Rosewell Park Memorial Institute-1640 (RPMI-1640 media, spinned at 30 rpm for 5 minutes. Microplate well was incubated for 24 and 48 hours in 37° C. After 24 hours, each well that will be read at 24 hour were added with 50 μl solution containing 5mg/ml MTT reagent in phosphate buffer saline (PBS solutions, then reincubated for 4 hours in CO2 10% and 37° C. Colorometric assay with MTT was used to evaluate viability of the cells population after 24 hours. Then, each well were added with 50 μl dimethyl sulfoxide (DMSO and reincubated for 5 minutes in 37° C. the wells were read using Elisa reader in 620 nm wave length. Same steps were done for the wells that will be read in 48 hours. Each data were tabulated and analyzed using independent T-test with significance of 5%. Results: This study showed that the percentage of living fibroblast after exposure to 316L stainless steel reconstruction plate was 61.58% after 24 hours and 62

  7. Lateral flow assays

    NARCIS (Netherlands)

    Posthuma-Trumpie, G.A.; Amerongen, van A.

    2012-01-01

    A simple version of immunochemical-based methods is the Lateral Flow Assay (LFA). It is a dry chemistry technique (reagents are included); the fluid from the sample runs through a porous membrane (often nitrocellulose) by capillary force. Typically the membrane is cut as a strip of 0.5*5 cm. In most

  8. Microchemiluminescent assay system

    Energy Technology Data Exchange (ETDEWEB)

    Kiel, J.L.

    1986-04-09

    The patent concerns a microchemiluminescent assay system, which can be used to detect ionizing radiation, heat or specific substances. The method involves the use of a complex formed from serum albumin and a luminescer which, in the presence of ionizing radiation (heat, or a specific analyte), will emit light in an amount proportional to the amount of radiation, etc. (U.K.).

  9. (MTT) dye reduction assay.

    African Journals Online (AJOL)

    to inhibit proliferation of HeLa cells was determined using the 3443- dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) dye reduction assay. Extracts from roots of Agathisanthemum bojeri, Synaptolepis kirkii and Zanha africana and the leaf extract of Physalis peruviana at a concentration of 10 pg/ml inhibited cell ...

  10. Hyaluronic Acid Assays

    DEFF Research Database (Denmark)

    Itenov, Theis S; Kirkby, Nikolai S; Bestle, Morten H

    2015-01-01

    BACKGROUD: Hyaluronic acid (HA) is proposed as a marker of functional liver capacity. The aim of the present study was to compare a new turbidimetric assay for measuring HA with the current standard method. METHODS: HA was measured by a particle-enhanced turbidimetric immunoassay (PETIA) and enzyme...

  11. FLUIDICS DEVICE FOR ASSAY

    DEFF Research Database (Denmark)

    2007-01-01

    The present invention relates to a device for use in performing assays on standard laboratory solid supports whereon chemical entities are attached. The invention furthermore relates to the use of such a device and a kit comprising such a device. The device according to the present invention is a...

  12. MyPlate Daily Checklist

    Science.gov (United States)

    ... Price Tag Read the Food Label Kitchen Timesavers Cooking for Your Family Tasty & Low-Cost Recipes Sample 2-Week Menus Resources for Professionals MyPlate Tip Sheets Print Materials Infographics 5 Ways ...

  13. License plate recognition (phase B).

    Science.gov (United States)

    2010-06-01

    License Plate Recognition (LPR) technology has been used for off-line automobile enforcement purposes. The technology has seen mixed success with correct reading rate as high as 60 to 80% depending on the specific application and environment. This li...

  14. Armor Plate Surface Roughness Measurements

    National Research Council Canada - National Science Library

    Stanton, Brian; Coburn, William; Pizzillo, Thomas J

    2005-01-01

    ...., surface texture and coatings) that could become important at high frequency. We measure waviness and roughness of various plates to know the parameter range for smooth aluminum and rolled homogenous armor (RHA...

  15. The optimal condition of performing MTT assay for the determination of radiation sensitivity

    International Nuclear Information System (INIS)

    Hong, Semie; Kim, Il Han

    2001-01-01

    The measurement of radiation survival using a clonogenic assay, the established standard, can be difficult and time consuming. In this study, We have used the MTT assay, based on the reduction of a tetrazolium salt to a purple formazan precipitate by living cells, as a substitution for clonogenic assay and have examined the optimal condition for performing this assay in determination of radiation sensitivity. Four human cancer cell lines - PCI-1, SNU-1066, NCI-H63O and RKO cells have been used. For each cell line, a clonogenic assay and a MTT assay using Premix WST-1 solution, which is one of the tetrazolium salts and does not require washing or solubilization of the precipitate were carried out after irradiation of 0, 2, 4, 6, 8, 10 Gy, For clonogenic assay, cells in 25 cm 2 flasks were irradiated after overnight incubation and the resultant colonies containing more than 50 cells were scored after culturing the cells for 10-14 days, For MTT assay, the relationship between absorbance and cell number, optimal seeding cell number, and optimal timing of assay was determined. Then, MTT assay was performed when the irradiated cells had regained exponential growth or when the non-irradiated cells had undergone four or more doubling times. There was minimal variation in the values gained from these two methods with the standard deviation generally less than 5%, and there were no statistically significant differences between two methods according to t-test in low radiation dose (below 6 Gy). The regression analyses showed high linear correlation with the R 2 value of 0.975-0.992 between data from the two different methods. The optimal cell numbers for MTT assay were found to be dependent on plating efficiency of used cell line. Less than 300 cells/well were appropriate for cells with high plating efficiency (more than 30%). For cells with low plating efficiency (less than 30%), 500 cells/well or more were appropriate for assay. The optimal time for MTT assay was alter 6

  16. Rhodium platings – experimental study

    OpenAIRE

    Rudolf, R.; Budić, B.; Stamenković, D.; Čolić, M.; Ivanič, A.; Kosec, B.

    2013-01-01

    Modern rhodium plating solutions are based on either sulphate or phosphate. Although in theory there are four possible combinations, in practice only three different rhodium electrolytes are used. These are based on dilutions of rhodium sulphate or phosphate concentrates with added sulphuric or phosphoric acid. These processes are be discussed in this paper with a demonstration of Rh platings in the Slovenian firm Zlatarna Celje d.d.

  17. Rhodium platings – experimental study

    Directory of Open Access Journals (Sweden)

    R. Rudolf

    2013-07-01

    Full Text Available Modern rhodium plating solutions are based on either sulphate or phosphate. Although in theory there are four possible combinations, in practice only three different rhodium electrolytes are used. These are based on dilutions of rhodium sulphate or phosphate concentrates with added sulphuric or phosphoric acid. These processes are be discussed in this paper with a demonstration of Rh platings in the Slovenian firm Zlatarna Celje d.d.

  18. Assay of free urinary cortisol with inverted phase liquid chromatography and radiocompetition

    International Nuclear Information System (INIS)

    Franck, C.; Patricot, M.C.; Mathian, B.; Revol, A.

    1984-01-01

    The authors present an assay method for free urinary cortisol using inverted phase liquid chromatography for purification and radiocompetition for the assay. The usual values for adult men and women were established. The results of 150 routine assays were compared with those of 17 OHCS revealing the value of this assay [fr

  19. Whole blood microculture assay of human lymphocyte function.

    Science.gov (United States)

    Pauly, J L; Han, T

    1976-11-01

    A whole blood microculture assay is described for measuring lymphocyte reactivity to mitogenic and antigenic stimulants. This assay employs heparinized whole blood, serum-free culture medium, microtiter plates, and a Multiple Automated Sample Harvester (MASH). When this assay is compared to other leukocyte assays, its major advantages include (1) the utilization of fewer lymphocytes per microculture, thuus reducing the amount of blood required per test while increasing the number of test agents and replicate cultures which can be employed in any given experiment; (2) the conservation of mitogens, antigens, drugs, enzymes, hormones, lymphokines, and other test agents, some of which are either expensive of difficult to prepare in large quantities; (3) the elimination of lymphocyte isolation and purification procedures which may disrupt the relative proportion of T cells, B cells and antigen-processing cells; and (4) the application of an automated harvester which simplifies and expedites procedures required for processing cells for liquid scintillation counting.

  20. A novel behavioral assay for measuring cold sensation in mice.

    Directory of Open Access Journals (Sweden)

    Daniel S Brenner

    Full Text Available Behavioral models of cold responses are important tools for exploring the molecular mechanisms of cold sensation. To complement the currently cold behavioral assays and allow further studies of these mechanisms, we have developed a new technique to measure the cold response threshold, the cold plantar assay. In this assay, animals are acclimated on a glass plate and a cold stimulus is applied to the hindpaw through the glass using a pellet of compressed dry ice. The latency to withdrawal from the cooled glass is used as a measure of the cold response threshold of the rodents, and the dry ice pellet provides a ramping cold stimulus on the glass that allows the correlation of withdrawal latency values to rough estimates of the cold response threshold temperature. The assay is highly sensitive to manipulations including morphine-induced analgesia, Complete Freund's Adjuvant-induced inflammatory allodynia, and Spinal Nerve Ligation-induced neuropathic allodynia.

  1. A novel behavioral assay for measuring cold sensation in mice.

    Science.gov (United States)

    Brenner, Daniel S; Golden, Judith P; Gereau, Robert W

    2012-01-01

    Behavioral models of cold responses are important tools for exploring the molecular mechanisms of cold sensation. To complement the currently cold behavioral assays and allow further studies of these mechanisms, we have developed a new technique to measure the cold response threshold, the cold plantar assay. In this assay, animals are acclimated on a glass plate and a cold stimulus is applied to the hindpaw through the glass using a pellet of compressed dry ice. The latency to withdrawal from the cooled glass is used as a measure of the cold response threshold of the rodents, and the dry ice pellet provides a ramping cold stimulus on the glass that allows the correlation of withdrawal latency values to rough estimates of the cold response threshold temperature. The assay is highly sensitive to manipulations including morphine-induced analgesia, Complete Freund's Adjuvant-induced inflammatory allodynia, and Spinal Nerve Ligation-induced neuropathic allodynia.

  2. Immunoradiometric assay for cytomegalovirus-specific IgG antibodies

    International Nuclear Information System (INIS)

    Klapper, P.E.; Cleator, G.M.; Prinja-Wolks, D.; Morris, D.J.

    1990-01-01

    An immunoradiometric assay (radio-immunosorbent test; RIST) for the detection of IgG antibodies to human herpesvirus 4 [human cytomegalovirus (CMV)] has been developed. The technique utilizes CMV antigen passively adsorbed to a polyvinyl microtitration plate and a radiolabelled murine monoclonal anti-human IgG antibody to detect binding of human antibody to the 'solid phase' reagent. The assay was optimized, and its specifity confirmed by testing paired acute and convalescent sera from patients with acute CMV or other human herpesvirus infections. To determine the assay's sensitivity 1433 blood donor sera were examined. The RIST was more sensitive than a standard complement fixation (CFT). Use of a monoclonal anti-human IgG antibody in the RIST reduced non-specific binding to the control uninfected cell antigen such that blood donor sera could be tested in the assay using only a CMV antigen without generating an unacceptable false positive rate. (author). 23 refs.; 1 tab

  3. Development of a solid-phase assay for measurement of proteolytic enzyme activity

    International Nuclear Information System (INIS)

    Varani, J.; Johnson, K.; Kaplan, J.

    1980-01-01

    A solid-phase, plate assay was developed for the measurement of proteolytic enzyme activity. In this assay procedure, radiolabeled substrates were dried onto the surface of microtiter wells. Following drying, the wells were washed two times with saline to remove the nonadherent substrate. When proteolytic enzymes were added to the wells, protein hydrolysis occurred, releasing radioactivity into the supernatant fluid. The amount of protein hydrolysis that occurred was reflected by the amount of radioactivity in the supernatant fluid. When 125 I-hemoglobin was used as the substrate, it was as susceptible to hydrolysis by trypsin in the solid-phase assay as it was in solution in a standard assay procedure. Protease activity from a variety of sources (including from viable cells as well as from extracellular sources) were also able to hydrolyze the hemoglobin on the plate. 125 I-Labeled serum albumen, fibrinogen, and rat pulmonary basement membrane were also susceptible to hydrolysis by trypsin in the solid phase. When [ 14 C]elastin was dried onto the plate, it behaved in a similar manner to elastin in solution. It was resistant to hydrolysis by nonspecific proteases such as trypsin and chymotrypsin but was highly susceptible to hydrolysis by elastase. The solid-phase plate assay has several features which recommended it for routine use. It is as sensitive as standard tube assays (and much more sensitive than routinely used colormetric assays). It is quick and convenient; there are no precipitation, centrifugation, or filtration steps. In addition, very small volumes of radioactive wastes are generated. Another advantage of the solid-phase plate assay is the resistance of the dried substrates to spontaneous breakdown and to microbial contamination. Finally, this assay is suitable for use with viable cells as well as for extracellular proteases

  4. Biocompatibility of Bespoke 3D-Printed Titanium Alloy Plates for Treating Acetabular Fractures

    Directory of Open Access Journals (Sweden)

    Xuezhi Lin

    2018-01-01

    Full Text Available Treatment of acetabular fractures is challenging, not only because of its complicated anatomy but also because of the lack of fitting plates. Personalized titanium alloy plates can be fabricated by selective laser melting (SLM but the biocompatibility of these three-dimensional printing (3D-printed plates remains unknown. Plates were manufactured by SLM and their cytocompatibility was assessed by observing the metabolism of L929 fibroblasts incubated with culture medium extracts using a CCK-8 assay and their morphology by light microscopy. Allergenicity was tested using a guinea pig maximization test. In addition, acute systemic toxicity of the 3D-printed plates was determined by injecting extracts from the implants into the tail veins of mice. Finally, the histocompatibility of the plates was investigated by implanting them into the dorsal muscles of rabbits. The in vitro results suggested that cytocompatibility of the 3D-printed plates was similar to that of conventional plates. The in vivo data also demonstrated histocompatibility that was comparable between the two manufacturing techniques. In conclusion, both in vivo and in vitro experiments suggested favorable biocompatibility of 3D-printed titanium alloy plates, indicating that it is a promising option for treatment of acetabular fractures.

  5. Radioreceptor assay for oxyphenonium

    International Nuclear Information System (INIS)

    Ensing, K.; Zeeuw, R.A. de

    1984-01-01

    The development of a radioreceptor assay for the quaternary anticholinergic drug, oxyphenonium, in plasma is reported. It is based on competition between this drug and 3 H-dexetimide for binding to muscarinic receptors. After ion pair extraction and reextraction, the drug can be determined in plasma at concentrations down to a value of 100 pg/ml. This permits pharmacokinetic studies to be made after inhalation of oxyphenonium. (author)

  6. Dual isotope assays

    International Nuclear Information System (INIS)

    Smith, G.F.W.; Stevens, R.A.J.; Jacoby, B.

    1980-01-01

    Dual isotope assays for thyroid function are performed by carrying out a radio-immunoassay for two of thyroxine (T4), tri-iodothyronine (T3), thyroid stimulating hormone (TSH), and thyroxine binding globulin (TBG), by a method wherein a version of one of the thyroid components, preferably T4 or T3 is labelled with Selenium-75 and the version of the other thyroid component is labelled with a different radionuclide, preferably Iodine-125. (author)

  7. Crustal Structure of the Tengchong Intra-plate Volcanic Area

    Science.gov (United States)

    Qian, Rongyi; Tong, Vincent C. H.

    2015-09-01

    We here provide an overview of our current understanding of the crustal structure of Tengchong in southwest China, a key intra-plate volcanic area along the Himalayan geothermal belt. Given that there is hitherto a lack of information about the near-surface structure of intra-plate volcanic areas, we present the first seismic reflection and velocity constraints on the shallow crust between intra-plate volcanoes. Our near-surface seismic images reveal the existence of dome-shaped seismic reflectors (DSRs) in the shallow crust between intra-plate volcanic clusters in Tengchong. The two DSRs are both ~2 km wide, and the shallowest parts of the DSRs are found at the depth of 200-300 m. The velocity model shows that the shallow low-velocity layer (<4 km/s) is anomalously thick (~1 km) in the region where the DSRs are observed. The presence of DSRs indicates significant levels of intra-plate magmatism beneath the along-axis gap separating two volcano clusters. Along-axis gaps between volcano clusters are therefore not necessarily an indicator of lower levels of magmatism. The seismic images obtained in this technically challenging area for controlled-source seismology allow us to conclude that shallow crustal structures are crucial for understanding the along-axis variations of magmatism and hydrothermal activities in intra-plate volcanic areas.

  8. Mathematics revealed

    CERN Document Server

    Berman, Elizabeth

    1979-01-01

    Mathematics Revealed focuses on the principles, processes, operations, and exercises in mathematics.The book first offers information on whole numbers, fractions, and decimals and percents. Discussions focus on measuring length, percent, decimals, numbers as products, addition and subtraction of fractions, mixed numbers and ratios, division of fractions, addition, subtraction, multiplication, and division. The text then examines positive and negative numbers and powers and computation. Topics include division and averages, multiplication, ratios, and measurements, scientific notation and estim

  9. A High Sensitivity Micro Format Chemiluminescence Enzyme Inhibition Assay for Determination of Hg(II

    Directory of Open Access Journals (Sweden)

    Kanchanmala Deshpande

    2010-06-01

    Full Text Available A highly sensitive and specific enzyme inhibition assay based on alcohol oxidase (AlOx and horseradish peroxidase (HRP for determination of mercury Hg(II in water samples has been presented. This article describes the optimization and miniaturization of an enzymatic assay using a chemiluminescence reaction. The analytical performance and detection limit for determination of Hg(II was optimized in 96 well plates and further extended to 384 well plates with a 10-fold reduction in assay volume. Inhibition of the enzyme activity by dissolved Hg(II was found to be linear in the range 5–500 pg.mL−1 with 3% CVin inter-batch assay. Due to miniaturization of assay in 384 well plates, Hg(II was measurable as low as 1 pg.mL−1 within15 min. About 10-fold more specificity of the developed assay for Hg(II analysis was confirmed by challenging with interfering divalent metal ions such as cadmium Cd(II and lead Pb(II. Using the proposed assay we could successfully demonstrate that in a composite mixture of Hg(II, Cd(II and Pb(II, inhibition by each metal ion is significantly enhanced in the presence of the others. Applicability of the proposed assay for the determination of the Hg(II in spiked drinking and sea water resulted in recoveries ranging from 100–110.52%.

  10. Modeling RERTR experimental fuel plates using the PLATE code

    International Nuclear Information System (INIS)

    Hayes, S.L.; Meyer, M.K.; Hofman, G.L.; Snelgrove, J.L.; Brazener, R.A.

    2003-01-01

    Modeling results using the PLATE dispersion fuel performance code are presented for the U-Mo/Al experimental fuel plates from the RERTR-1, -2, -3 and -5 irradiation tests. Agreement of the calculations with experimental data obtained in post-irradiation examinations of these fuels, where available, is shown to be good. Use of the code to perform a series of parametric evaluations highlights the sensitivity of U-Mo dispersion fuel performance to fabrication variables, especially fuel particle shape and size distributions. (author)

  11. Plating on some difficult-to-plate metals and alloys

    International Nuclear Information System (INIS)

    Dini, J.W.; Johnson, H.R.

    1980-02-01

    Electrodeposition of coatings on metals such as beryllium, beryllium-copper, Kovar, lead, magnesium, thorium, titanium, tungsten, uranium, zirconium, and their alloys can be problematic. This is due in most cases to a natural oxide surface film that readily reforms after being removed. The procedures we recommend for plating on these metals rely on replacing the oxide film with a displacement coating, or etching to allow mechanical keying between the substrate and plated deposit. The effectiveness of the procedures is demonstrated by interface bond strengths found in ring-shear and conical-head tensile tests

  12. Automation of the anthrone assay for carbohydrate concentration determinations.

    Science.gov (United States)

    Turula, Vincent E; Gore, Thomas; Singh, Suddham; Arumugham, Rasappa G

    2010-03-01

    Reported is the adaptation of a manual polysaccharide assay applicable for glycoconjugate vaccines such as Prevenar to an automated liquid handling system (LHS) for improved performance. The anthrone assay is used for carbohydrate concentration determinations and was scaled to the microtiter plate format with appropriate mixing, dispensing, and measuring operations. Adaptation and development of the LHS platform was performed with both dextran polysaccharides of various sizes and pneumococcal serotype 6A polysaccharide (PnPs 6A). A standard plate configuration was programmed such that the LHS diluted both calibration standards and a test sample multiple times with six replicate preparations per dilution. This extent of replication minimized the effect of any single deviation or delivery error that might have occurred. Analysis of the dextran polymers ranging in size from 214 kDa to 3.755 MDa showed that regardless of polymer chain length the hydrolysis was complete, as evident by uniform concentration measurements. No plate positional absorbance bias was observed; of 12 plates analyzed to examine positional bias the largest deviation observed was 0.02% percent relative standard deviation (%RSD). The high purity dextran also afforded the opportunity to assess LHS accuracy; nine replicate analyses of dextran yielded a mean accuracy of 101% recovery. As for precision, a total of 22 unique analyses were performed on a single lot of PnPs 6A, and the resulting variability was 2.5% RSD. This work demonstrated the capability of a LHS to perform the anthrone assay consistently and a reduced assay cycle time for greater laboratory capacity.

  13. Transfer plate radioassay using cell monolayers to detect anti-cell surface antibodies synthesized by lymphocyte hybridomas

    International Nuclear Information System (INIS)

    Schneider, M.D.; Eisenbarth, G.S.

    1979-01-01

    A solid phase [ 125 I] Protein A radioassay for anti-cell surface antibodies is described, which employs target cell monolayers cultured on fenestrated polyvinyl chloride 96-well plates ('transfer plates'). The calibrated aperture in the bottom of each well is small enough to retain fluid contents by surface tension during monolayer growth, but also permits fluid to enter the wells when transfer plate are lowered into receptacles containing washing buffer on test sera. To assay for antibodies directed against target cell surface antigens, transfer plates bearing monolayers are inserted into microculture plates with corresponding 96-well geometry, thereby simultaneously sampling 96 wells. This assay allows rapid screening of hundreds of hybrid cell colonies for production of antibodies with desired tissue specificity. (Auth.)

  14. Electroless metal plating of plastics

    International Nuclear Information System (INIS)

    Krause, L.J.

    1986-01-01

    The product of an electroless plating process is described for plating at least one main group metal directly on a surface of a polymeric substrate comprising the steps of forming a nonaqueous solution containing a metallic salt of an alkali metal in a positive valence state and at least one main group metal in a negative valence state, the main group metal being selected from the group consisting of Ge, Sn, Pb, As, Sb, Bi, Si and Te, selecting an aromatic polymeric substrate reducible by the solublized salt and resistant to degration during the reaction, and carrying out a redox reaction between the salt in solution and the substrate by contacting the solution with the substrate for a sufficient time to oxidize and deposit the main group metal in elemental form to produce a plated substrate. The product is characterized by the plated metal being directly on the surface of the polymeric substrate and the alkali metal being retained in the plated substrate with the substrate being negatively charged with electrons transferred from the main group metal during the redox reaction

  15. Soft Plate and Impact Tectonics

    Science.gov (United States)

    Tikoff, Basil

    In the field of tectonics, most of our ideas are published in journals. This is not true of other fields, such as history, in which ideas are primarily published in books. Within my own field of structural geology, I can recall only one book, Strain Fades by E. Hansen (Springer-Verlag, 1971), which presents a new idea in book form. However, even this book is more useful for its philosophical approach and particular methodology of determining directions of folding, than for its overarching idea.Enter Soft Plate and Impact Tectonics, a new book with an interesting hypothesis that has been informally discussed in the geoscience community: A fundamental tenet of plate tectonics is incorrect—namely, that the plates are rigid. This assertion is evident when looking at any mountain range, and is perhaps most clearly stated in Molnar [1988].

  16. NF-kappaB specifically activates BMP-2 gene expression in growth plate chondrocytes in vivo and in a chondrocyte cell line in vitro.

    Science.gov (United States)

    Feng, Jian Q; Xing, Lianping; Zhang, Jiang-Hong; Zhao, Ming; Horn, Diane; Chan, Jeannie; Boyce, Brendan F; Harris, Stephen E; Mundy, Gregory R; Chen, Di

    2003-08-01

    Bone morphogenetic protein-2 (BMP-2) regulates growth plate chondrogenesis during development and postnatal bone growth, but the control mechanisms of BMP-2 expression in growth plate chondrocytes are unknown. Here we have used both in vitro and in vivo approaches to demonstrate that transcription factor, NF-kappaB, regulates BMP-2 gene expression in chondrocytes. Two putative NF-kappaB response elements were found in the -2712/+165 region of the BMP-2 gene. Cotransfection of mutant I-kappaBalpha expression plasmids with BMP-2 promoter-luciferase reporters into TMC-23 chondrocyte cell line suppressed BMP-2 transcription. Mutations in NF-kappaB response elements in the BMP-2 gene lead to decreases in BMP-2 promoter activity. Electrophoretic mobility shift assay using nuclear extracts from TMC-23 chondrocytic cells revealed that the NF-kappaB subunits p50 and p65 bound to the NF-kappaB response elements of the BMP-2 gene. Thus, NF-kappaB may positively regulate BMP-2 gene transcription. Consistent with these findings, expression of BMP-2 mRNA was significantly reduced in growth plate chondrocytes in NF-kappaB p50/p52 dKO mice, which associated with decreased numbers of 5-bromo-2'-deoxyuridine (BrdUrd)-positive cells in the proliferating zone of growth plate in these mice. Therefore, in postnatal growth plate chondrocytes, expression of BMP-2 is regulated by NF-kappaB, which may play an important role in chondrogenesis.

  17. Semiclassical theory of plate vibrations

    International Nuclear Information System (INIS)

    Bogomolny, E.; Hugues, E.

    1996-11-01

    The bi-harmonic equation of flexural vibrations of elastic plates is studied by a semiclassical method which can easily be generalized for other models of wave propagation. The surface and perimeter terms of the asymptotic number of levels are derived exactly. The next constant term is also derived. A semiclassical approximation of the quantization condition is obtained. A Berry-Tabor formula and a Gutzwiller trace formula are deduced for the integrable and chaotic cases respectively. From 600 eigenvalues of a clamped stadium plate obtained by a specially developed numerical algorithm, the trace formula is assessed, looking at its Fourier transform compared with the membrane case. (author)

  18. Radiorespirometic assay device

    International Nuclear Information System (INIS)

    Levin, G.V.; Straat, P.A.

    1981-01-01

    A radiorespirometic assay device is described in which the presence of microorganisms in a sample is determined by placing the sample in contact with a metabolisable radioactive labelled substrate, collecting any gas evolved, exposing a photosensitive material to the gas and determining if a spot is produced on the material. A spot indicates the presence of radioactivity showing that the substrate has been metabolized by a microorganism. Bacteria may be detected in body fluids, hospital operating rooms, water, food, cosmetics and drugs. (U.K.)

  19. Radon assay for SNO+

    Energy Technology Data Exchange (ETDEWEB)

    Rumleskie, Janet [Laurentian University, Greater Sudbury, Ontario (Canada)

    2015-12-31

    The SNO+ experiment will study neutrinos while located 6,800 feet below the surface of the earth at SNOLAB. Though shielded from surface backgrounds, emanation of radon radioisotopes from the surrounding rock leads to back-grounds. The characteristic decay of radon and its daughters allows for an alpha detection technique to count the amount of Rn-222 atoms collected. Traps can collect Rn-222 from various positions and materials, including an assay skid that will collect Rn-222 from the organic liquid scintillator used to detect interactions within SNO+.

  20. Probe colorimeter for quantitating enzyme-linked immunosorbent assays and other colorimetric assays performed with microplates.

    Science.gov (United States)

    Ackerman, S B; Kelley, E A

    1983-03-01

    The performance of a fiberoptic probe colorimeter (model PC800; Brinkmann Instruments, Inc., Westbury, N.Y.) for quantitating enzymatic or colorimetric assays in 96-well microtiter plates was compared with the performances of a spectrophotometer (model 240; Gilford Instrument Laboratories, Inc., Oberlin, Ohio) and a commercially available enzyme immunoassay reader (model MR590; Dynatech Laboratories, Inc., Alexandria, Va.). Alkaline phosphatase-p-nitrophenyl phosphate in 3 M NaOH was used as the chromophore source. Six types of plates were evaluated for use with the probe colorimeter; they generated reproducibility values (100% coefficient of variation) ranging from 91 to 98% when one individual made 24 independent measurements on the same dilution of chromophore on each plate. Eleven individuals each performed 24 measurements with the colorimeter on either a visually light (absorbance of 0.10 at 420 nm) or a dark (absorbance of 0.80 at 420 nm) dilution of chromophore; reproducibilities averaged 87% for the light dilution and 97% for the dark dilution. When one individual measured the same chromophore sample at least 20 times in the colorimeter, in the spectrophotometer or in the enzyme immunoassay reader, reproducibility for each instrument was greater than 99%. Measurements of a dilution series of chromophore in a fixed volume indicated that the optical responses of each instrument were linear in a range of 0.05 to 1.10 absorbance units.

  1. Improving shuffler assay accuracy

    International Nuclear Information System (INIS)

    Rinard, P.M.

    1995-01-01

    Drums of uranium waste should be disposed of in an economical and environmentally sound manner. The most accurate possible assays of the uranium masses in the drums are required for proper disposal. The accuracies of assays from a shuffler are affected by the type of matrix material in the drums. Non-hydrogenous matrices have little effect on neutron transport and accuracies are very good. If self-shielding is known to be a minor problem, good accuracies are also obtained with hydrogenous matrices when a polyethylene sleeve is placed around the drums. But for those cases where self-shielding may be a problem, matrices are hydrogenous, and uranium distributions are non-uniform throughout the drums, the accuracies are degraded. They can be greatly improved by determining the distributions of the uranium and then applying correction factors based on the distributions. This paper describes a technique for determining uranium distributions by using the neutron count rates in detector banks around the waste drum and solving a set of overdetermined linear equations. Other approaches were studied to determine the distributions and are described briefly. Implementation of this correction is anticipated on an existing shuffler next year

  2. Competitive protein binding assay

    International Nuclear Information System (INIS)

    Kaneko, Toshio; Oka, Hiroshi

    1975-01-01

    The measurement of cyclic GMP (cGMP) by competitive protein binding assay was described and discussed. The principle of binding assay was represented briefly. Procedures of our method by binding protein consisted of preparation of cGMP binding protein, selection of 3 H-cyclic GMP on market, and measurement procedures. In our method, binding protein was isolated from the chrysalis of silk worm. This method was discussed from the points of incubation medium, specificity of binding protein, the separation of bound cGMP from free cGMP, and treatment of tissue from which cGMP was extracted. cGMP existing in the tissue was only one tenth or one scores of cGMP, and in addition, cGMP competed with cGMP in binding with binding protein. Therefore, Murad's technique was applied to the isolation of cGMP. This method provided the measurement with sufficient accuracy; the contamination by cAMP was within several per cent. (Kanao, N.)

  3. The effects of emitter-tied field plates on lateral PNP ionizing radiation response

    International Nuclear Information System (INIS)

    Barnaby, H.J.; Schrimpf, R.D.; Cirba, C.R.; Pease, R.L.; Fleetwood, D.M.; Kosier, S.L.

    1998-03-01

    Radiation response comparisons of lateral PNP bipolar technologies reveal that device hardening may be achieved by extending the emitter contact over the active base. The emitter-tied field plate suppresses recombination of carriers with interface traps

  4. The Earth's Mantle Is Solid: Teachers' Misconceptions About the Earth and Plate Tectonics.

    Science.gov (United States)

    King, Chris

    2000-01-01

    Discusses the misconceptions revealed by the teachers' answers and outlines more accurate answers and explanations based on established evidence and uses these to provide a more complete understanding of plate tectonic process and the structure of Earth. (Author/YDS)

  5. The tectonic plates are moving!

    CERN Document Server

    Livermore, Roy

    2018-01-01

    Written in a witty and informal style, this book explains modern plate tectonics in a non-technical manner, showing not only how it accounts for phenomena such as great earthquakes, tsunamis, and volcanic eruptions, but also how it controls conditions at the Earth’s surface, including global geography and climate, making it suitable for life. The book presents the advances that have been made since the establishment of plate tectonics in the 1960s, highlighting, on the fiftieth anniversary of the theory, the contributions of a small number of scientists who have never been widely recognized for their discoveries. Beginning with the publication of a short article in Nature by Vine and Matthews, the book traces the development of plate tectonics through two generations of the theory. First-generation plate tectonics covers the exciting scientific revolution of the 1960s, its heroes, and its villains. The second generation includes the rapid expansions in sonar, satellite, and seismic technologies during the 1...

  6. License plate recognition using DTCNNs

    NARCIS (Netherlands)

    ter Brugge, M.H; Stevens, J.H; Nijhuis, J.A G; Spaanenburg, L; Tavsanonoglu, V

    1998-01-01

    Automatic license plate recognition requires a series of complex image processing steps. For practical use, the amount of data to he processed must be minimized early on. This paper shows that the computationally most intensive steps can be realized by DTCNNs. Moreover; high-level operations like

  7. Hydroelasticity of a Floating Plate

    DEFF Research Database (Denmark)

    Chen, X.; Jensen, Jørgen Juncher; Cui, W.

    2003-01-01

    The membrane forces are included in the hydroelastic analysis of a floating plate undergoing large vertical deflections in regular monochromatic multidirectional waves. The first-order vertical displacements induced by the linear wave exciting forces are calculated by the mode expansion method in...

  8. Stress state in perforated plates

    International Nuclear Information System (INIS)

    Visner, J.

    1977-01-01

    The method is described of photoelastic measurement of stress concentration factors (s.c.f) in plates perforated by a square, triangular and diagonal grid of circular holes and loaded by uniaxial or biaxial tensile stress. A loading equipment which was developed and its modifications are described. Stress concentration factors found are compared with theoretical and experimental results given in references. (author)

  9. Regulators of growth plate maturation

    NARCIS (Netherlands)

    Emons, Joyce Adriana Mathilde

    2010-01-01

    Estrogen is known to play an important role in longitudinal bone growth and growth plate maturation, but the mechanism by which estrogens exert their effect is not fully understood. In this thesis this role is further explored. Chapter 1 contains a general introduction to longitudinal bone growth

  10. Automatic cell cloning assay for determining the clonogenic capacity of cancer and cancer stem-like cells.

    Science.gov (United States)

    Fedr, Radek; Pernicová, Zuzana; Slabáková, Eva; Straková, Nicol; Bouchal, Jan; Grepl, Michal; Kozubík, Alois; Souček, Karel

    2013-05-01

    The clonogenic assay is a well-established in vitro method for testing the survival and proliferative capability of cells. It can be used to determine the cytotoxic effects of various treatments including chemotherapeutics and ionizing radiation. However, this approach can also characterize cells with different phenotypes and biological properties, such as stem cells or cancer stem cells. In this study, we implemented a faster and more precise method for assessing the cloning efficiency of cancer stem-like cells that were characterized and separated using a high-speed cell sorter. Cell plating onto a microplate using an automatic cell deposition unit was performed in a single-cell or dilution rank mode by the fluorescence-activated cell sorting method. We tested the new automatic cell-cloning assay (ACCA) on selected cancer cell lines and compared it with the manual approach. The obtained results were also compared with the results of the limiting dilution assay for different cell lines. We applied the ACCA to analyze the cloning capacity of different subpopulations of prostate and colon cancer cells based on the expression of the characteristic markers of stem (CD44 and CD133) and cancer stem cells (TROP-2, CD49f, and CD44). Our results revealed that the novel ACCA is a straightforward approach for determining the clonogenic capacity of cancer stem-like cells identified in both cell lines and patient samples. Copyright © 2013 International Society for Advancement of Cytometry.

  11. Revealing Rembrandt

    Directory of Open Access Journals (Sweden)

    Andrew J Parker

    2014-04-01

    Full Text Available The power and significance of artwork in shaping human cognition is self-evident. The starting point for our empirical investigations is the view that the task of neuroscience is to integrate itself with other forms of knowledge, rather than to seek to supplant them. In our recent work, we examined a particular aspect of the appreciation of artwork using present-day functional magnetic resonance imaging (fMRI. Our results emphasised the continuity between viewing artwork and other human cognitive activities. We also showed that appreciation of a particular aspect of artwork, namely authenticity, depends upon the co-ordinated activity between the brain regions involved in multiple decision making and those responsible for processing visual information. The findings about brain function probably have no specific consequences for understanding how people respond to the art of Rembrandt in comparison with their response to other artworks. However, the use of images of Rembrandt’s portraits, his most intimate and personal works, clearly had a significant impact upon our viewers, even though they have been spatially confined to the interior of an MRI scanner at the time of viewing. Neuroscientific studies of humans viewing artwork have the capacity to reveal the diversity of human cognitive responses that may be induced by external advice or context as people view artwork in a variety of frameworks and settings.

  12. Identification of Lactobacillus delbrueckii and Streptococcus thermophilus Strains Present in Artisanal Raw Cow Milk Cheese Using Real-time PCR and Classic Plate Count Methods.

    Science.gov (United States)

    Stachelska, Milena A

    2017-12-04

    The aim of this paper was to detect Lactobacillus delbrueckii and Streptococcus thermophilus using real-time quantitative PCR assay in 7-day ripening cheese produced from unpasteurised milk. Real-time quantitative PCR assays were designed to identify and enumerate the chosen species of lactic acid bacteria (LAB) in ripened cheese. The results of molecular quantification and classic bacterial enumeration showed a high level of similarity proving that DNA extraction was carried out in a proper way and that genomic DNA solutions were free of PCR inhibitors. These methods revealed the presence of L. delbrueckii and S. thermophilus. The real-time PCR enabled quantification with a detection of 101-103 CFU/g of product. qPCR-standard curves were linear over seven log units down to 101 copies per reaction; efficiencies ranged from 77.9% to 93.6%. Cheese samples were analysed with plate count method and qPCR in parallel. Compared with the classic plate count method, the newly developed qPCR method provided faster and species specific identification of two dairy LAB and yielded comparable quantitative results.

  13. Natural Convective Heat Transfer from Narrow Plates

    CERN Document Server

    Oosthuizen, Patrick H

    2013-01-01

    Natural Convective Heat Transfer from Narrow Plates deals with a heat transfer situation that is of significant practical importance but which is not adequately dealt with in any existing textbooks or in any widely available review papers. The aim of the book is to introduce the reader to recent studies of natural convection from narrow plates including the effects of plate edge conditions, plate inclination, thermal conditions at the plate surface and interaction of the flows over adjacent plates. Both numerical and experimental studies are discussed and correlation equations based on the results of these studies are reviewed.

  14. Application of biodegradable plates for treating pediatric mandibular fractures.

    Science.gov (United States)

    An, Jingang; Jia, Pengcheng; Zhang, Yi; Gong, Xi; Han, Xiaodong; He, Yang

    2015-05-01

    We assessed the clinical results of a biodegradable plate system for the internal fixation of mandibular fractures in children, and observed the imaging features of fracture healing and bone changes around the biodegradable plates and screws during follow-up. We enrolled 39 patients (22 male, 17 female, average age 4 years 10 months) with different mandibular fractures. We used 2.0-mm resorbable plates to repair the fractures. Postoperative follow-up ranged from 6 months to 5 years; average follow-up was 1 year 2 months. The outcome measures identified and assessed included facial symmetry, mouth opening, occlusal relationship, infection, nonunion, malunion, and plate dehiscence. We fixed 42 fractures with 43 resorbable plates; the fracture site of one patient (aged 11 years 3 months) was fixed with two plates. Two patients developed small fistulas at the intraoral incision 2 months after surgery; the fistulas healed after 1 month without special treatment. In the other patients, the incision healed well, there was facial symmetry, mouth opening was >35 mm, and occlusion was good. Follow-up computed tomography examination data were available for 20 cases, and revealed different degrees of radiolucency indicating that osteolysis had occurred. Radiolucency was observed around the resorbable plates 1 month after the surgery. The extent and depth of the radiolucent region were obvious within 1 year of surgery. In the second year, there were obvious repairs, with the bony defect areas becoming shallower. After 2 years, the bony defect areas had almost disappeared. Biodegradable fixation devices are safe and efficient for treating pediatric mandibular fractures. Osteolysis commonly follows biodegradable fixation of pediatric mandibular fractures, and has no adverse effect on fracture healing. Copyright © 2015 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.

  15. An acoustic prion assay

    Directory of Open Access Journals (Sweden)

    Gordon Hayward

    2016-12-01

    Full Text Available An acoustic prion assay has been demonstrated for sheep brain samples. Only five false positives and no false negatives were observed in a test of 45 positive and 45 negative samples. The acoustic prion sensor was constructed using a thickness shear mode quartz resonator coated with a covalently bound recombinant prion protein. The characteristic indicator of a scrapie infected sheep brain sample was an observed shoulder in the frequency decrease in response to a sample.The response of the sensor aligns with a conformational shift in the surface protein and with the propagation mechanism of the disease. This alignment is evident in the response timing and shape, dependence on concentration, cross species behaviour and impact of blood plasma. This alignment is far from sufficient to prove the mechanism of the sensor but it does offer the possibility of a rapid and inexpensive additional tool to explore prion disease. Keywords: Prions, Thickness shear mode quartz sensor

  16. Assay of oestrogen

    International Nuclear Information System (INIS)

    Edwards, J.C.

    1981-01-01

    A particular problem with the direct radioimmunoassay of unconjugated oestriol in pregnancy is caused by the increased amount of steroid-binding proteins present in pregnancy serum and plasma. The steroid-binding proteins react with oestriol and 125 I-labelled oestriol during the assay procedure and the steroid-protein bound 125 I-labelled oestriol is precipitated along with the antibody-bound 125 I-labelled oestriol by the ammonium sulphate solution separation system. A novel method is described whereby progesterone (1-20 μg/ml) is used to block the action of steroid-binding proteins in pregnancy serum and plasma samples, thus minimizing interference in a direct radioimmunoassay for unconjugated oestriol using a specific anti-oestriol serum. (U.K.)

  17. Medically Relevant Assays with a Simple Smartphone and Tablet Based Fluorescence Detection System

    OpenAIRE

    Wargocki, Piotr; Deng, Wei; Anwer, Ayad G.; Goldys, Ewa M.

    2015-01-01

    Cell phones and smart phones can be reconfigured as biomedical sensor devices but this requires specialized add-ons. In this paper we present a simple cell phone-based portable bioassay platform, which can be used with fluorescent assays in solution. The system consists of a tablet, a polarizer, a smart phone (camera) and a box that provides dark readout conditions. The assay in a well plate is placed on the tablet screen acting as an excitation source. A polarizer on top of the well plate s...

  18. Fully automated synthesis of (phosphopeptide arrays in microtiter plate wells provides efficient access to protein tyrosine kinase characterization

    Directory of Open Access Journals (Sweden)

    Goldstein David J

    2005-01-01

    Full Text Available Abstract Background Synthetic peptides have played a useful role in studies of protein kinase substrates and interaction domains. Synthetic peptide arrays and libraries, in particular, have accelerated the process. Several factors have hindered or limited the applicability of various techniques, such as the need for deconvolution of combinatorial libraries, the inability or impracticality of achieving full automation using two-dimensional or pin solid phases, the lack of convenient interfacing with standard analytical platforms, or the difficulty of compartmentalization of a planar surface when contact between assay components needs to be avoided. This paper describes a process for synthesis of peptides and phosphopeptides on microtiter plate wells that overcomes previous limitations and demonstrates utility in determination of the epitope of an autophosphorylation site phospho-motif antibody and utility in substrate utilization assays of the protein tyrosine kinase, p60c-src. Results The overall reproducibility of phospho-peptide synthesis and multiplexed EGF receptor (EGFR autophosphorylation site (pY1173 antibody ELISA (9H2 was within 5.5 to 8.0%. Mass spectrometric analyses of the released (phosphopeptides showed homogeneous peaks of the expected molecular weights. An overlapping peptide array of the complete EGFR cytoplasmic sequence revealed a high redundancy of 9H2 reactive sites. The eight reactive phospopeptides were structurally related and interestingly, the most conserved antibody reactive peptide motif coincided with a subset of other known EGFR autophosphorylation and SH2 binding motifs and an EGFR optimal substrate motif. Finally, peptides based on known substrate specificities of c-src and related enzymes were synthesized in microtiter plate array format and were phosphorylated by c-Src with the predicted specificities. The level of phosphorylation was proportional to c-Src concentration with sensitivities below 0.1 Units of

  19. Subduction Drive of Plate Tectonics

    Science.gov (United States)

    Hamilton, W. B.

    2003-12-01

    Don Anderson emphasizes that plate tectonics is self-organizing and is driven by subduction, which rights the density inversion generated as oceanic lithosphere forms by cooling of asthenosphere from the top. The following synthesis owes much to many discussions with him. Hinge rollback is the key to kinematics, and, like the rest of actual plate behavior, is incompatible with bottom-up convection drive. Subduction hinges (which are under, not in front of, thin leading parts of arcs and overriding plates) roll back into subducting plates. The Pacific shrinks because bounding hinges roll back into it. Colliding arcs, increasing arc curvatures, back-arc spreading, and advance of small arcs into large plates also require rollback. Forearcs of overriding plates commonly bear basins which preclude shortening of thin plate fronts throughout periods recorded by basin strata (100 Ma for Cretaceous and Paleogene California). This requires subequal rates of advance and rollback, and control of both by subduction. Convergence rate is equal to rates of rollback and advance in many systems but is greater in others. Plate-related circulation probably is closed above 650 km. Despite the popularity of concepts of plumes from, and subduction into, lower mantle, there is no convincing evidence for, and much evidence against, penetration of the 650 in either direction. That barrier not only has a crossing-inhibiting negative Clapeyron slope but also is a compositional boundary between fractionated (not "primitive"), sluggish lower mantle and fertile, mobile upper mantle. Slabs sink more steeply than they dip. Slabs older than about 60 Ma when their subduction began sink to, and lie down on and depress, the 650-km discontinuity, and are overpassed, whereas younger slabs become neutrally buoyant in mid-upper mantle, into which they are mixed as they too are overpassed. Broadside-sinking old slabs push all upper mantle, from base of oceanic lithosphere down to the 650, back under

  20. EMPIRICAL STUDY OF CAR LICENSE PLATES RECOGNITION

    Directory of Open Access Journals (Sweden)

    Nasa Zata Dina

    2015-01-01

    Full Text Available The number of vehicles on the road has increased drastically in recent years. The license plate is an identity card for a vehicle. It can map to the owner and further information about vehicle. License plate information is useful to help traffic management systems. For example, traffic management systems can check for vehicles moving at speeds not permitted by law and can also be installed in parking areas to se-cure the entrance or exit way for vehicles. License plate recognition algorithms have been proposed by many researchers. License plate recognition requires license plate detection, segmentation, and charac-ters recognition. The algorithm detects the position of a license plate and extracts the characters. Various license plate recognition algorithms have been implemented, and each algorithm has its strengths and weaknesses. In this research, I implement three algorithms for detecting license plates, three algorithms for segmenting license plates, and two algorithms for recognizing license plate characters. I evaluate each of these algorithms on the same two datasets, one from Greece and one from Thailand. For detecting li-cense plates, the best result is obtained by a Haar cascade algorithm. After the best result of license plate detection is obtained, for the segmentation part a Laplacian based method has the highest accuracy. Last, the license plate recognition experiment shows that a neural network has better accuracy than other algo-rithm. I summarize and analyze the overall performance of each method for comparison.

  1. Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices.

    OpenAIRE

    Christensen, G D; Simpson, W A; Younger, J J; Baddour, L M; Barrett, F F; Melton, D M; Beachey, E H

    1985-01-01

    The adherence of coagulase-negative staphylococci to smooth surfaces was assayed by measuring the optical densities of stained bacterial films adherent to the floors of plastic tissue culture plates. The optical densities correlated with the weight of the adherent bacterial film (r = 0.906; P less than 0.01). The measurements also agreed with visual assessments of bacterial adherence to culture tubes, microtiter plates, and tissue culture plates. Selected clinical strains were passed through ...

  2. Earth's Decelerating Tectonic Plates

    Energy Technology Data Exchange (ETDEWEB)

    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.

  3. Silicon-micromachined microchannel plates

    CERN Document Server

    Beetz, C P; Steinbeck, J; Lemieux, B; Winn, D R

    2000-01-01

    Microchannel plates (MCP) fabricated from standard silicon wafer substrates using a novel silicon micromachining process, together with standard silicon photolithographic process steps, are described. The resulting SiMCP microchannels have dimensions of approx 0.5 to approx 25 mu m, with aspect ratios up to 300, and have the dimensional precision and absence of interstitial defects characteristic of photolithographic processing, compatible with positional matching to silicon electronics readouts. The open channel areal fraction and detection efficiency may exceed 90% on plates up to 300 mm in diameter. The resulting silicon substrates can be converted entirely to amorphous quartz (qMCP). The strip resistance and secondary emission are developed by controlled depositions of thin films, at temperatures up to 1200 deg. C, also compatible with high-temperature brazing, and can be essentially hydrogen, water and radionuclide-free. Novel secondary emitters and cesiated photocathodes can be high-temperature deposite...

  4. Mathematical methods for elastic plates

    CERN Document Server

    Constanda, Christian

    2014-01-01

    Mathematical models of deformation of elastic plates are used by applied mathematicians and engineers in connection with a wide range of practical applications, from microchip production to the construction of skyscrapers and aircraft. This book employs two important analytic techniques to solve the fundamental boundary value problems for the theory of plates with transverse shear deformation, which offers a more complete picture of the physical process of bending than Kirchhoff’s classical one.   The first method transfers the ellipticity of the governing system to the boundary, leading to singular integral equations on the contour of the domain. These equations, established on the basis of the properties of suitable layer potentials, are then solved in spaces of smooth (Hölder continuous and Hölder continuously differentiable) functions.   The second technique rewrites the differential system in terms of complex variables and fully integrates it, expressing the solution as a combination of complex ana...

  5. PLATE

    DEFF Research Database (Denmark)

    Kling, Joyce; Hjulmand, Lise-Lotte

    2008-01-01

    Copenhagen Business School (CBS) finds itself needing to address the issue of English-medium instruction for its increasing number of foreign exchange and full degree students. With internationalisation as a main pillar of the institution’s agenda, there are concerns whether the teaching faculty...... of academic teaching and lecturing into account....

  6. Electroless metal plating of plastics

    Science.gov (United States)

    Krause, L.J.

    1982-09-20

    Process for plating main group metals on aromatic polymers is carried out by the use of a nonaqueous solution of a salt of an alkali metal in a positive valence state and a main group metal in a negative valence state with contact between the solution and polymer providing a redox reaction causing the deposition of the main group metal and the reduction of the polymer. Products from the process exhibit useful decorative and electrical properties.

  7. A tandem parallel plate analyzer

    International Nuclear Information System (INIS)

    Hamada, Y.; Fujisawa, A.; Iguchi, H.; Nishizawa, A.; Kawasumi, Y.

    1996-11-01

    By a new modification of a parallel plate analyzer the second-order focus is obtained in an arbitrary injection angle. This kind of an analyzer with a small injection angle will have an advantage of small operational voltage, compared to the Proca and Green analyzer where the injection angle is 30 degrees. Thus, the newly proposed analyzer will be very useful for the precise energy measurement of high energy particles in MeV range. (author)

  8. Curved Folded Plate Timber Structures

    OpenAIRE

    Buri, Hans Ulrich; Stotz, Ivo; Weinand, Yves

    2011-01-01

    This work investigates the development of a Curved Origami Prototype made with timber panels. In the last fifteen years the timber industry has developed new, large size, timber panels. Composition and dimensions of these panels and the possibility of milling them with Computer Numerical Controlled machines shows great potential for folded plate structures. To generate the form of these structures we were inspired by Origami, the Japanese art of paper folding. Common paper tessellations are c...

  9. PRETREATING URANIUM FOR METAL PLATING

    Science.gov (United States)

    Wehrmann, R.F.

    1961-05-01

    A process is given for anodically treating the surface of uranium articles, prior to metal plating. The metal is electrolyzed in an aqueous solution of about 10% polycarboxylic acid, preferably oxalic acid, from 1 to 5% by weight of glycerine and from 1 to 5% by weight of hydrochloric acid at from 20 to 75 deg C for from 30 seconds to 15 minutes. A current density of from 60 to 100 amperes per square foot is used.

  10. Embedded adhesive connection for laminated glass plates

    DEFF Research Database (Denmark)

    Hansen, Jens Zangenberg; Poulsen, S.H.; Bagger, A.

    2012-01-01

    The structural behavior of a new connection design, the embedded adhesive connection, used for laminated glass plates is investigated. The connection consists of an aluminum plate encapsulated in-between two adjacent triple layered laminated glass plates. Fastening between glass and aluminum...... usage in a design situation. The embedded connection shows promising potential as a future fastening system for load-carrying laminated glass plates....

  11. Modified tubularized incised plate urethroplasty

    Directory of Open Access Journals (Sweden)

    Shivaji Mane

    2013-01-01

    Full Text Available Aim: To share our experience of doing tubularized incised plate urethroplasty with modifications. Materials and Methods: This is a single surgeon personal series from 2004 to 2009. One hundred patients of distal hypospadias were subjected for Snodgrass urethroplasty with preputioplasty. The age range was 1 to 5 year with mean age of 2.7 years. Selection criteria were good urethral plate, without chordee and torsion needing complete degloving. Main technical modification from original Snodgrass procedure was spongioplasty, preputioplasty, and dorsal slit when inability to retract prepuce during surgery. Results: Average follow-up period is 23 months. Seven (7% patients developed fistula and one patient had complete preputial dehiscence. Phimosis developed in three (3% patients and required circumcision. Dorsal slit was required in seven patients. One patient developed meatal stenosis in postoperative period. All other patients are passing single urinary stream and have cosmesis that is acceptable. Conclusions: Modified tubularized incised plate urethroplasty with preputioplasty effectively gives cosmetically normal looking penis with low complications.

  12. Divertor plate for thermonuclear reactor

    International Nuclear Information System (INIS)

    Yamazaki, Seiichiro; Sato, Keisuke; Nishio, Satoshi.

    1993-01-01

    In a divertor plate for a thermonuclear reactor, adjacent cooling pipes are electrically insulated from each other and pipes made of a gradient functional material prepared by compositing ceramics having an insulation property and metals are metallurgically joined to at least one portion of each of the cooling pipes. Electric current caused upon occurrence of plasma disruption is interrupted by the insulation portion, so that a large circuit is not formed and electromagnetic force is decreased to such a extent that the divertor plate is not ruptured. Since a header of the cooling pipes can be installed at any optional position, the installation space can be reduced. Further, since inlet and exit collection headers can be disposed on both ends of the cooling pipes, it is possible to shorten the length of the cooling pipe of the divertor plate corresponded to high heat fluxes and reduce the pressure loss on the side of coolants to about 1/2. Further, turn back portions of small radius of curvature of the cooling pipes are eliminated to reduce the cost and extend the lifetime and, in addition, protection tiles can be attached easily. (N.H.)

  13. Secure matching of Dutch car license plates

    NARCIS (Netherlands)

    Sunil, A.B.; Erkiny, Z.; Veugenyz, T.

    2016-01-01

    License plate matching plays an important role in applications like law enforcement, traffic management and road pricing, where the plate is first recognized and then compared to a database of authorized vehicle registration plates. Unfortunately, there are several privacy related issues that should

  14. Moving Divertor Plates in a Tokamak

    International Nuclear Information System (INIS)

    Zweben, S.J.; Zhang, H.

    2009-01-01

    Moving divertor plates could help solve some of the problems of the tokamak divertor through mechanical ingenuity rather than plasma physics. These plates would be passively heated on each pass through the tokamak and cooled and reprocessed outside the tokamak. There are many design options using varying plate shapes, orientations, motions, coatings, and compositions

  15. Designing a licence plate for memorability

    NARCIS (Netherlands)

    Schraagen, J.M.C.; Dongen, C.J.G. van

    2005-01-01

    Good memorability of licence plates is important in those cases where licence plates are viewed for a brief period of time and the information is essential for police investigations. The purpose of the current study was to design a new Dutch licence plate that could be remembered well. A memory

  16. Strength of Ship Plates under Combined Loading

    DEFF Research Database (Denmark)

    Cui, W.; Wang, Y.; Pedersen, Preben Terndrup

    2002-01-01

    Strength of ship plates plays a significant role in the ultimate strength analysis of ship structures. In recent years several authors have proposed simplified analytical methods to calculate the ultimate strength of unstiffened plates. The majority of these investigations deal with plates subjec...

  17. Strength of ship plates under combined loading

    DEFF Research Database (Denmark)

    Cui, Weiching; Wang, Yongjun; Pedersen, Preben Terndrup

    2000-01-01

    Strength of ship plates plays a significant role for the ultimate strength analysis of ship structures. In recent years several authors have proposed simplified methods to calculate the ultimate strength of unstiffened plates. The majority of these investigations deal with plates subjected to lon...

  18. Operational test of bonded magnetocaloric plates

    DEFF Research Database (Denmark)

    Bahl, Christian; Navickaité, Kristina; Neves Bez, Henrique

    2017-01-01

    Bonded plates made by hot pressing La0.85Ce0.15Fe11.25Mn0.25Si1.5Hy particles and resin have been tested as active magnetic regenerators in a small scale magnetocaloric device. Firstly the plates were carefully characterised magnetically and thermally. The plates were prepared with 5 wt% resin...

  19. Moving Divertor Plates in a Tokamak

    Energy Technology Data Exchange (ETDEWEB)

    S.J. Zweben, H. Zhang

    2009-02-12

    Moving divertor plates could help solve some of the problems of the tokamak divertor through mechanical ingenuity rather than plasma physics. These plates would be passively heated on each pass through the tokamak and cooled and reprocessed outside the tokamak. There are many design options using varying plate shapes, orientations, motions, coatings, and compositions.

  20. Variables affecting viral plaque formation in microculture plaque assays using homologous antibody in a liquid overlay.

    Science.gov (United States)

    Randhawa, A S; Stanton, G J; Green, J A; Baron, S

    1977-05-01

    A liquid antibody microculture plaque assay and the variables that govern its effectiveness are described. The assay is based on the principle that low concentrations of homologous antibody can inhibit secondary plaque formation without inhibiting formation of primary plaques. Thus, clear plaques that followed a linear dose response were produced. The assay was found to be more rapid, less cumbersome, and less expensive than assays using agar overlays and larger tissue culture plates. It was reproducible, quantitative, and had about the same sensitivity as the agar overlay technique in measuring infectious coxsackievirus type B-3. It was more sensitive in assaying adenovirus type 3 and Western equine encephalomyelitis, vesicular stomatitis, Semliki forest, Sendai, Sindbis, and Newcastle disease viruses than were liquid, carboxymethylcellulose, and methylcellulose microculture plaque assays. The variables influencing sensitivity and accuracy, as determined by using coxsackievirus type B-3, were: (i) the inoculum volume of virus; (ii) the incubation period of virus; and (iii) the incubation temperature.

  1. Hydrodynamics of a three-dimensional self-propelled flexible plate

    Science.gov (United States)

    Ryu, Jaeha; Sung, Hyung Jin

    2017-11-01

    A three-dimensional self-propelled flexible plate in a quiescent flow was simulated using the immersed boundary method. The clamped leading edge of the flexible plate was forced into a vertical oscillation, while free to move horizontally. To reveal the hydrodynamics of the plate, the averaged cruising speed (UC) , the input power (P) , and the swimming efficiency (η) were analyzed as a function of the bending rigidity (γ) and the flapping frequency (f) . The velocity field around the plate and the exerted force on the plate were demonstrated to find out the dynamic interaction between the plate and the surrounding fluid. The kinematics of the plate, the maximum angle of attack (ϕmax) , and the mean effective length (Leff) were examined accounting for the hydrodynamics of the self-propelled flexible plate. The vortical structures around the plate were visualized, and the influence of the tip vortex on the swimming efficiency was explored qualitatively and quantitatively. This work was supported by the Creative Research Initiatives (No. 2017-013369) program of the National Research Foundation of Korea (MSIP).

  2. Plate tectonics, habitability and life

    Science.gov (United States)

    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

  3. Medically relevant assays with a simple smartphone and tablet based fluorescence detection system.

    Science.gov (United States)

    Wargocki, Piotr; Deng, Wei; Anwer, Ayad G; Goldys, Ewa M

    2015-05-20

    Cell phones and smart phones can be reconfigured as biomedical sensor devices but this requires specialized add-ons. In this paper we present a simple cell phone-based portable bioassay platform, which can be used with fluorescent assays in solution. The system consists of a tablet, a polarizer, a smart phone (camera) and a box that provides dark readout conditions. The assay in a well plate is placed on the tablet screen acting as an excitation source. A polarizer on top of the well plate separates excitation light from assay fluorescence emission enabling assay readout with a smartphone camera. The assay result is obtained by analysing the intensity of image pixels in an appropriate colour channel. With this device we carried out two assays, for collagenase and trypsin using fluorescein as the detected fluorophore. The results of collagenase assay with the lowest measured concentration of 3.75 µg/mL and 0.938 µg in total in the sample were comparable to those obtained by a microplate reader. The lowest measured amount of trypsin was 930 pg, which is comparable to the low detection limit of 400 pg for this assay obtained in a microplate reader. The device is sensitive enough to be used in point-of-care medical diagnostics of clinically relevant conditions, including arthritis, cystic fibrosis and acute pancreatitis.

  4. Medically Relevant Assays with a Simple Smartphone and Tablet Based Fluorescence Detection System

    Directory of Open Access Journals (Sweden)

    Piotr Wargocki

    2015-05-01

    Full Text Available Cell phones and smart phones can be reconfigured as biomedical sensor devices but this requires specialized add-ons. In this paper we present a simple cell phone-based portable bioassay platform, which can be used with fluorescent assays in solution. The system consists of a tablet, a polarizer, a smart phone (camera and a box that provides dark readout conditions. The assay in a well plate is placed on the tablet screen acting as an excitation source. A polarizer on top of the well plate separates excitation light from assay fluorescence emission enabling assay readout with a smartphone camera. The assay result is obtained by analysing the intensity of image pixels in an appropriate colour channel. With this device we carried out two assays, for collagenase and trypsin using fluorescein as the detected fluorophore. The results of collagenase assay with the lowest measured concentration of 3.75 µg/mL and 0.938 µg in total in the sample were comparable to those obtained by a microplate reader. The lowest measured amount of trypsin was 930 pg, which is comparable to the low detection limit of 400 pg for this assay obtained in a microplate reader. The device is sensitive enough to be used in point-of-care medical diagnostics of clinically relevant conditions, including arthritis, cystic fibrosis and acute pancreatitis.

  5. Width of electromagnetic wave instability spectrum in tungsten plate

    International Nuclear Information System (INIS)

    Rinkevich, A.B.

    1995-01-01

    Based on the study of high-frequency signal modulation and spectrum analysis of the envelope a measurement of spectrum width for electromagnetic wave instability was carried out under conditions of current pulse action on tungsten plate in magnetic field. The existence of amplitude-frequency wave modulation was revealed. The width of current disturbance spectrum in a specimen was evaluated. Current disturbances are shown to cause the instability of electromagnetic wave. 11 refs.; 6 figs

  6. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci.

    Science.gov (United States)

    Stepanović, Srdjan; Vuković, Dragana; Hola, Veronika; Di Bonaventura, Giovanni; Djukić, Slobodanka; Cirković, Ivana; Ruzicka, Filip

    2007-08-01

    The details of all steps involved in the quantification of biofilm formation in microtiter plates are described. The presented protocol incorporates information on assessment of biofilm production by staphylococci, gained both by direct experience as well as by analysis of methods for assaying biofilm production. The obtained results should simplify quantification of biofilm formation in microtiter plates, and make it more reliable and comparable among different laboratories.

  7. Free vibration analysis of corroded steel plates

    Energy Technology Data Exchange (ETDEWEB)

    Eslami-Majd, Alireza; Rahbar-Ranji, Ahmad [AmirKabir University of Technology, Tehran (Iran, Islamic Republic of)

    2014-06-15

    Vibration analysis of unstiffened/stiffened plates has long been studied due to its importance in the design and condition assessments of ship and offshore structures. Corrosion is inevitable in steel structures and has been so far considered in strength analysis of structures. We studied the free vibration of pitted corroded plates with simply supported boundary conditions. Finite element analysis, with ABAQUS, was used to determine the natural frequencies and mode shapes of corroded plates. Influential parameters including plate aspect ratio, degree of pit, one-sided/both-sided corroded plate, and different corrosion patterns were investigated. By increasing the degree of corrosion, reduction of natural frequency increases. Plate aspect ratio and plate dimensions have no influence on reduction of natural frequency. Different corrosion patterns on the surface of one-sided corroded plates have little influence on reduction of natural frequency. Ratio of pit depth over plate thickness has no influence on the reduction of natural frequency. The reduction of natural frequency in both-sided corroded plates is higher than one-sided corroded plates with the same amount of total corrosion loss. Mode shapes of vibration would change due to corrosion, except square mode shapes.

  8. Bottom nozzle for nuclear reactor fuel assembly having an adaptor plate and a coupled filtration plate

    International Nuclear Information System (INIS)

    Verdier, M.; Mortgat, R.

    1992-01-01

    The bottom nozzle includes an adaptor plate with openings to allow the passage of water and a filtration plate with small holes. The openings in the adaptor plate are symmetrical with regard to medians and diagonals. Within each zone, some of the openings are rectangular and some may be circular. The small holes in the filtration plate coincide with the rectangular openings in the adaptor plate

  9. METHOD OF MAKING AND PLATE CONNECTION

    Science.gov (United States)

    Patriarca, P.; Shubert, C.E.; Slaughter, G.M.

    1963-02-26

    This invention relates to a method of joining, by welding or brazing, a tube to a plate, particularly where the tube and the plate are of different thickness or have different thermal conductivities and are subject to high temperatures. In this method the tube is inserted in the core of a plate containing an annular groove in its back face concentric with the bore and in communication with the bore. One end of the tube is welded substantially flush with the front end of the plate. Brazing alloy is placed in the groove on the back face of the plate and heat is applied to the plate and tube to melt the brazing alloy to permit the alloy to flow into the bore and bond the tube to the plate. (AEC)

  10. PWR integral tie plate and locking mechanism

    International Nuclear Information System (INIS)

    Flora, B.S.; Osborne, J.L.

    1980-01-01

    A locking mechanism for securing an upper tie plate to the tie rods of a nuclear fuel bundle is described. The mechanism includes an upper tie plate assembly and locking sleeves fixed to the ends of the tie rods. The tie plate is part of the upper tie plate assembly and is secured to the fuel bundle by securing the entire upper tie plate assembly to the locking sleeves fixed to the tie rods. The assembly includes, in addition to the tie plate, locking nuts for engaging the locking sleeves, retaining sleeves to operably connect the locking nuts to the assembly, a spring biased reaction plate to restrain the locking nuts in the locked position and a means to facilitate the removal of the entire assembly as a unit from the fuel bundle

  11. Primary DNA damage in chrome-plating workers.

    Science.gov (United States)

    Gambelunghe, A; Piccinini, R; Ambrogi, M; Villarini, M; Moretti, M; Marchetti, C; Abbritti, G; Muzi, G

    2003-06-30

    In order to evaluate the primary DNA damage due to occupational exposure to chromium (VI), DNA strand-breaks and apoptosis in peripheral lymphocytes were measured in a group of 19 chrome-plating workers. DNA strand-breaks was assessed by alkaline (pH>13) single-cell microgel electrophoresis ('comet') assay, while apoptosis was measured by flow-cytometry after propidium iodide staining of the cells. Concentrations of chromium in urine, erythrocytes and lymphocytes were investigated as biological indicators of exposure. A group of 18 hospital workers (control group I) and another 20 university personnel (control group II) without exposure to chromium were also studied as controls. The results of the study show that chrome-plating workers have higher levels of chromium in urine, erythrocytes and lymphocytes than unexposed workers. Comet tail moment values, assumed as index of DNA damage, are increased in chromium-exposed workers and results are significantly correlated to chromium lymphocyte concentrations. No difference emerged in the percentage of apoptotic nuclei in exposed and unexposed workers. The study confirms that measurements of chromium in erythrocytes and lymphocytes may provide useful information about recent and past exposure to hexavalent chromium at the workplace. The increase in DNA strand-breaks measured by comet assay suggests this test is valid for the biological monitoring of workers exposed to genotoxic compounds such as chromium (VI).

  12. Optimization of Phenotyping Assays for the Model Monocot Setaria viridis.

    Science.gov (United States)

    Acharya, Biswa R; Roy Choudhury, Swarup; Estelle, Aiden B; Vijayakumar, Anitha; Zhu, Chuanmei; Hovis, Laryssa; Pandey, Sona

    2017-01-01

    Setaria viridis (green foxtail) is an important model plant for the study of C4 photosynthesis in panicoid grasses, and is fast emerging as a system of choice for the study of plant development, domestication, abiotic stress responses and evolution. Basic research findings in Setaria are expected to advance research not only in this species and its close relative S. italica (foxtail millet), but also in other panicoid grasses, many of which are important food or bioenergy crops. Here we report on the standardization of multiple growth and development assays for S. viridis under controlled conditions, and in response to several phytohormones and abiotic stresses. We optimized these assays at three different stages of the plant's life: seed germination and post-germination growth using agar plate-based assays, early seedling growth and development using germination pouch-based assays, and adult plant growth and development under environmentally controlled growth chambers and greenhouses. These assays will be useful for the community to perform large scale phenotyping analyses, mutant screens, comparative physiological analysis, and functional characterization of novel genes of Setaria or other related agricultural crops. Precise description of various growth conditions, effective treatment conditions and description of the resultant phenotypes will help expand the use of S. viridis as an effective model system.

  13. Optimization of Phenotyping Assays for the Model Monocot Setaria viridis

    Directory of Open Access Journals (Sweden)

    Biswa R. Acharya

    2017-12-01

    Full Text Available Setaria viridis (green foxtail is an important model plant for the study of C4 photosynthesis in panicoid grasses, and is fast emerging as a system of choice for the study of plant development, domestication, abiotic stress responses and evolution. Basic research findings in Setaria are expected to advance research not only in this species and its close relative S. italica (foxtail millet, but also in other panicoid grasses, many of which are important food or bioenergy crops. Here we report on the standardization of multiple growth and development assays for S. viridis under controlled conditions, and in response to several phytohormones and abiotic stresses. We optimized these assays at three different stages of the plant’s life: seed germination and post-germination growth using agar plate-based assays, early seedling growth and development using germination pouch-based assays, and adult plant growth and development under environmentally controlled growth chambers and greenhouses. These assays will be useful for the community to perform large scale phenotyping analyses, mutant screens, comparative physiological analysis, and functional characterization of novel genes of Setaria or other related agricultural crops. Precise description of various growth conditions, effective treatment conditions and description of the resultant phenotypes will help expand the use of S. viridis as an effective model system.

  14. Vortex Dynamics around Pitching Plates

    Science.gov (United States)

    2014-04-29

    electrical signals are A/D converted in an ATI NetBox interface and recorded using a Java application, and are filtered in three steps. The first is a low...the plate while staying attached to the corners of the leading edge. During this process, a second vortex loop, created by the quick angular ...is a spike in CL centered around t = 0 due to non-circulatory6 effects from the angular acceleration of the wing. The amplitude of the peak is

  15. Micro plate fission chamber development

    International Nuclear Information System (INIS)

    Wang Mei; Wen Zhongwei; Lin Jufang; Jiang Li; Liu Rong; Wang Dalun

    2014-01-01

    To conduct the measurement of neutron flux and the fission rate distribution at several position in assemblies, the micro plate fission chamber was designed and fabricated. Since the requirement of smaller volume and less structure material was taken into consideration, it is convinient, commercial and practical to use fission chamber to measure neutron flux in specific condition. In this paper, the structure of fission chamber and process of fabrication were introduced and performance test result was presented. The detection efficiency is 91.7%. (authors)

  16. Non-cyanide silver plating

    International Nuclear Information System (INIS)

    Dini, J.W.

    1995-01-01

    Lawrence Livermore National Laboratory (LLNL) and Technic, Inc. have entered into a CRADA (Cooperative Research and Development Agreement) with the goal of providing industry with an environmentally benign alternative to the presently used silver cyanide plating process. This project has been in place for about six months and results are quite promising. The main objective, that of deposition of deposits as thick as 125 um (5 mils), has been met. Property data such as stress and hardness have been obtained and the structure of the deposit has been analyzed via metallography and x-ray diffraction. These results will be presented in this paper, along with plans for future work

  17. Infection Assay of Cyst Nematodes on Arabidopsis Roots.

    Science.gov (United States)

    Bohlmann, Holger; Wieczorek, Krzysztof

    2015-09-20

    Plant parasitic nematodes are devastating pests on many crops. Juveniles (J2) of cyst nematodes invade the roots to induce a syncytium. This feeding site is their only source of nutrients. Male nematodes leave the roots after the fourth molt to mate with females. The females stay attached to their syncytia throughout their life and produce hundreds of eggs, which are contained in their bodies. When the females die their bodies form the cysts, which protect the eggs. Cysts can survive for many years in the soil until favorable conditions induce hatching of the juveniles. The beet cyst nematode Heterodera schachtii ( H. schachtii )is a pathogen of sugar beet ( Beta vulgaris ) but can also complete its life cycle on Arabidopsis roots growing on agar plates under sterile conditions. We present here protocols for a stock culture of H. schachtii and an infection assay on agar plates.

  18. Cell Painting, a high-content image-based assay for morphological profiling using multiplexed fluorescent dyes

    Science.gov (United States)

    Bray, Mark-Anthony; Singh, Shantanu; Han, Han; Davis, Chadwick T.; Borgeson, Blake; Hartland, Cathy; Kost-Alimova, Maria; Gustafsdottir, Sigrun M.; Gibson, Christopher C.; Carpenter, Anne E.

    2016-01-01

    In morphological profiling, quantitative data are extracted from microscopy images of cells to identify biologically relevant similarities and differences among samples based on these profiles. This protocol describes the design and execution of experiments using Cell Painting, a morphological profiling assay multiplexing six fluorescent dyes imaged in five channels, to reveal eight broadly relevant cellular components or organelles. Cells are plated in multi-well plates, perturbed with the treatments to be tested, stained, fixed, and imaged on a high-throughput microscope. Then, automated image analysis software identifies individual cells and measures ~1,500 morphological features (various measures of size, shape, texture, intensity, etc.) to produce a rich profile suitable for detecting subtle phenotypes. Profiles of cell populations treated with different experimental perturbations can be compared to suit many goals, such as identifying the phenotypic impact of chemical or genetic perturbations, grouping compounds and/or genes into functional pathways, and identifying signatures of disease. Cell culture and image acquisition takes two weeks; feature extraction and data analysis take an additional 1-2 weeks. PMID:27560178

  19. Influence of the wavelet order on proper damage location in plate structures

    Science.gov (United States)

    Pawlak, Zdzisław; Knitter-Piątkowska, Anna

    2018-01-01

    The rectangular thin plates were analyzed in the paper. The static response in plate structure subjected to the uniform load was derived by applying the finite element method. In the dynamic, experimental tests the accelerations were obtained with the use of modal hammer and DEWEsoft® software. Next, the analysis of the signal was carried out with the use of Discrete Wavelet Transform (DWT), provided that damage exists in the considered plate structure. It was assumed, that in the middle of the structure a certain area of the plate is thinner or there is a crack across the entire plate thickness. The aim of this work was to choose the appropriate wavelet order to reveal the localization of defect. The results of selected numerical example proved the efficiency of proposed approach.

  20. Sirt6 regulates postnatal growth plate differentiation and proliferation via Ihh signaling.

    Science.gov (United States)

    Piao, Jinying; Tsuji, Kunikazu; Ochi, Hiroki; Iwata, Munetaka; Koga, Daisuke; Okawa, Atsushi; Morita, Sadao; Takeda, Shu; Asou, Yoshinori

    2013-10-23

    Sirtuin 6 (Sirt6) is a mammalian homologue of NAD⁺-dependent histone deacetylase Sir2. Although Sirt6⁻/⁻ mice exhibit growth retardation, the role of Sirt6 in cartilage metabolism is unclear. The aim of this study was to investigate the Sirt6 signaling pathway in cartilage metabolism. Immunohistological evaluation of the tibial growth plate in Sirt6⁻/⁻ mice exhibited impaired proliferation and differentiation of chondrocytes, reduced expression of Indian hedgehog (Ihh), and a senescent phenotype. When Sirt6 was knocked down in chondrocytes in vitro, expression of Ihh and its downstream genes were reduced. Impaired differentiation by Sirt6 silencing was completely rescued by administration of a Hh signal agonist. When sirtuins were activated, chondrocyte differentiation was enhanced together with activation of Ihh signal, and these effects were abrogated by Sirt6 silencing. ChIP assay revealed the affinity of ATF4 to the Ihh promoter was markedly decreased by Sirt6 knockdown. These data indicate Sirt6 directly controls proliferation and differentiation of chondrocytes.

  1. Swath sonar mapping of Earth's submarine plate boundaries

    Science.gov (United States)

    Carbotte, S. M.; Ferrini, V. L.; Celnick, M.; Nitsche, F. O.; Ryan, W. B. F.

    2014-12-01

    The recent loss of Malaysia Airlines flight MH370 in an area of the Indian Ocean where less than 5% of the seafloor is mapped with depth sounding data (Smith and Marks, EOS 2014) highlights the striking lack of detailed knowledge of the topography of the seabed for much of the worlds' oceans. Advances in swath sonar mapping technology over the past 30 years have led to dramatic improvements in our capability to map the seabed. However, the oceans are vast and only an estimated 10% of the seafloor has been mapped with these systems. Furthermore, the available coverage is highly heterogeneous and focused within areas of national strategic priority and community scientific interest. The major plate boundaries that encircle the globe, most of which are located in the submarine environment, have been a significant focus of marine geoscience research since the advent of swath sonar mapping. While the location of these plate boundaries are well defined from satellite-derived bathymetry, significant regions remain unmapped at the high-resolutions provided by swath sonars and that are needed to study active volcanic and tectonic plate boundary processes. Within the plate interiors, some fossil plate boundary zones, major hotspot volcanoes, and other volcanic provinces have been the focus of dedicated research programs. Away from these major tectonic structures, swath mapping coverage is limited to sparse ocean transit lines which often reveal previously unknown deep-sea channels and other little studied sedimentary structures not resolvable in existing low-resolution global compilations, highlighting the value of these data even in the tectonically quiet plate interiors. Here, we give an overview of multibeam swath sonar mapping of the major plate boundaries of the globe as extracted from public archives. Significant quantities of swath sonar data acquired from deep-sea regions are in restricted-access international archives. Open access to more of these data sets would

  2. Upgrading well plates using open microfluidic patterning.

    Science.gov (United States)

    Berry, Samuel B; Zhang, Tianzi; Day, John H; Su, Xiaojing; Wilson, Ilham Z; Berthier, Erwin; Theberge, Ashleigh B

    2017-12-05

    Cellular communication between multiple cell types is a ubiquitous process that is responsible for vital physiological responses observed in vivo (e.g., immune response, organ function). Many in vitro coculture strategies have been developed, both in traditional culture and microscale systems, and have shown the potential to recreate some of the physiological behaviors of organs or groups of cells. A fundamental limitation of current systems is the difficulty of reconciling the additional engineering requirements for creating soluble factor signaling systems (e.g., segregated cell culture) with the use of well-characterized materials and platforms that have demonstrated successful results and biocompatibility in assays. We present a new open-microfluidic platform, the Monorail Device, that is placed in any existing well plate or Petri dish and enables patterning of segregated coculture regions, thereby allowing the direct upgrade of monoculture experiments into multiculture assays. Our platform patterns biocompatible hydrogel walls via microfluidic spontaneous capillary flow (SCF) along a rail insert set inside commercially available cultureware, creating customized pipette-accessible cell culture chambers that require fewer cells than standard macroscale culture. Importantly, the device allows the use of native surfaces without additional modification or treatments, while creating permeable dividers for the diffusion of soluble factors. Additionally, the ease of patterning afforded by our platform makes reconfiguration of the culture region as simple as changing the rail insert. We demonstrate the ability of the device to pattern flows on a variety of cell culture surfaces and create hydrogel walls in complex and precise shapes. We characterize the physical parameters that enable a reproducible SCF-driven flow and highlight specialized design features that increase the ease of use of the device and control of the open microfluidic flow. Further, we present the

  3. Silicon-micromachined microchannel plates

    International Nuclear Information System (INIS)

    Beetz, Charles P.; Boerstler, Robert; Steinbeck, John; Lemieux, Bryan; Winn, David R.

    2000-01-01

    Microchannel plates (MCP) fabricated from standard silicon wafer substrates using a novel silicon micromachining process, together with standard silicon photolithographic process steps, are described. The resulting SiMCP microchannels have dimensions of ∼0.5 to ∼25 μm, with aspect ratios up to 300, and have the dimensional precision and absence of interstitial defects characteristic of photolithographic processing, compatible with positional matching to silicon electronics readouts. The open channel areal fraction and detection efficiency may exceed 90% on plates up to 300 mm in diameter. The resulting silicon substrates can be converted entirely to amorphous quartz (qMCP). The strip resistance and secondary emission are developed by controlled depositions of thin films, at temperatures up to 1200 deg. C, also compatible with high-temperature brazing, and can be essentially hydrogen, water and radionuclide-free. Novel secondary emitters and cesiated photocathodes can be high-temperature deposited or nucleated in the channels or the first strike surface. Results on resistivity, secondary emission and gain are presented

  4. Stress analysis of plate-fin structures in recuperator

    International Nuclear Information System (INIS)

    Matsui, Shingo; Muto, Yasushi; Shiina, Yasuaki

    2001-01-01

    A high performance compact recuperator with 95% effectiveness is required to achieve a high thermal efficiency power generation of up to 50% in High Temperature Gas Cooled Reactor (HTGR) coupled with closed cycle helium gas turbine. Though a plate-fin type heat exchanger is proposed for this recuperator, much research and development works are needed to establish this high performance goal since there exists no state-of-the-art technology in such a high pressure and high temperature one. One of the important works is to establish the structural analysis and evaluation method in this plate-fin type heat exchanger. This paper describes the results of stress analysis of the plate-fin structure under the internal pressure as the first step of this work. First, the modeling of a unit plate-fin structure for the analysis was examined and a three layers model was confirmed to be most adequate. The stress distribution within the structure was clarified by using this model. Second, the three layers model was simplified to one layer model with sufficient accuracy. By using this model, both the effects of an inclined angle of fin and a thickness of separate on the strength were examined parametrically. Under the relevant design conditions, it was revealed that the optimum inclined angle of fin locates in the neighborhood of 76 degree rather than most difficult fabrication angle 90 degree and there is possibility to adopt thinner thickness than 0.5 mm in the current design. (author)

  5. Active structural health monitoring of composite plates and sandwiches

    Directory of Open Access Journals (Sweden)

    Sadílek P.

    2013-12-01

    Full Text Available The aim of presented work is to design, assemble and test a functional system, that is able to reveal damage from impact loading. This is done by monitoring of change of spectral characteristics on a damaged structure that is caused by change of mechanical properties of material or by change of structure’s geometry. Excitation and monitoring of structures was done using piezoelectric patches. Unidirectional composite plate was tested for eigenfrequencies using chirp signal. The eigenfrequencies were compared to results from experiments with an impact hammer and consequently with results from finite element method. Same method of finding eigenfrequencies was used on a different unidirectional composite specimen. Series of impacts were performed. Spectrum of eigenfrequencies was measured on undamaged plate and then after each impact. Measurements of the plate with different level of damage were compared. Following experiments were performed on sandwich materials where more different failures may happen. Set of sandwich beams (cut out from one plate made of two outer composite layers and a foam core was investigated and subjected to several impacts. Several samples were impacted in the same manner to get comparable results. The impacts were performed with growing impact energy.

  6. Buckling analysis for axially compressed flat plates, structural sections, and stiffened plates reinforced with laminated composites

    Science.gov (United States)

    Viswanathan, A. V.; Soong, T.; Miller, R. E., Jr.

    1971-01-01

    A classical buckling analysis is developed for stiffened, flat plates composed of a series of linked plate and beam elements. Plates are idealized as multilayered orthotropic elements. Structural beads and lips are idealized as beams. The loaded edges of the stiffened plate are simply-supported and the conditions at the unloaded edges can be prescribed arbitrarily. The plate and beam elements are matched along their common junctions for displacement continuity and force equilibrium in an exact manner. Offsets between elements are considered in the analysis. Buckling under uniaxial compressive load for plates, sections, and stiffened plates is investigated. Buckling loads are the lowest of all possible general and local failure modes, and the mode shape is used to determine whether buckling is a local or general instability. Numerical correlations with existing analysis and test data for plates, sections, and stiffened plates including boron-reinforced structures are discussed. In general correlations are reasonably good.

  7. Bone plate composed of a ternary nano-hydroxyapatite/polyamide 66/glass fiber composite: biomechanical properties and biocompatibility.

    Science.gov (United States)

    Qiao, Bo; Li, Jidong; Zhu, Qingmao; Guo, Shuquan; Qi, Xiaotong; Li, Weichao; Wu, Jun; Liu, Yang; Jiang, Dianming

    2014-01-01

    An ideal bone plate for internal fixation of bone fractures should have good biomechanical properties and biocompatibility. In this study, we prepared a new nondegradable bone plate composed of a ternary nano-hydroxyapatite/polyamide 66/glass fiber (n-HA/PA66/GF) composite. A breakage area on the n-HA/PA66/GF plate surface was characterized by scanning electron microscopy. Its mechanical properties were investigated using bone-plate constructs and biocompatibility was evaluated in vitro using bone marrow-derived mesenchymal stem cells. The results confirmed that adhesion between the n-HA/PA66 matrix and the glass fibers was strong, with only a few fibers pulled out at the site of breakage. Fractures fixed by the n-HA/PA66/GF plate showed lower stiffness and had satisfactory strength compared with rigid fixation using a titanium plate. Moreover, the results with regard to mesenchymal stem cell morphology, MTT assay, Alizarin Red S staining, enzyme-linked immunosorbent assay, and reverse transcription polymerase chain reaction for alkaline phosphatase and osteocalcin showed that the n-HA/PA66/GF composite was suitable for attachment and proliferation of mesenchymal stem cells, and did not have a negative influence on matrix mineralization or osteogenic differentiation of mesenchymal stem cells. These observations indicate that the n-HA/PA66/GF plate has good biomechanical properties and biocompatibility, and may be considered a new option for internal fixation in orthopedic surgery.

  8. Nanoparticle-assay marker interaction: effects on nanotoxicity assessment

    International Nuclear Information System (INIS)

    Zhao, Xinxin; Xiong, Sijing; Huang, Liwen Charlotte; Ng, Kee Woei; Loo, Say Chye Joachim

    2015-01-01

    Protein-based cytotoxicity assays such as lactate dehydrogenase (LDH) and tumor necrosis factor-alpha (TNF-α) are commonly used in cytotoxic evaluation of nanoparticles (NPs) despite numerous reports on possible interactions with protein markers in these assays that can confound the results obtained. In this study, conventional cytotoxicity assays where assay markers may (LDH and TNF- α) or may not (PicoGreen and WST-8) come into contact with NPs were used to evaluate the cytotoxicity of NPs. The findings revealed selective interactions between negatively charged protein assay markers (LDH and TNF- α) and positively charged ZnO NPs under abiotic conditions. The adsorption and interaction with these protein assay markers were strongly influenced by surface charge, concentration, and specific surface area of the NPs, thereby resulting in less than accurate cytotoxic measurements, as observed from actual cell viability measurements. An improved protocol for LDH assay was, therefore, proposed and validated by eliminating any effects associated with protein–particle interactions. In view of this, additional measures and precautions should be taken when evaluating cytotoxicity of NPs with standard protein-based assays, particularly when they are of opposite charges

  9. Nanoparticle-assay marker interaction: effects on nanotoxicity assessment

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Xinxin; Xiong, Sijing; Huang, Liwen Charlotte; Ng, Kee Woei, E-mail: kwng@ntu.edu.sg; Loo, Say Chye Joachim, E-mail: joachimloo@ntu.edu.sg [Nanyang Technological University, School of Materials Science and Engineering (Singapore)

    2015-01-15

    Protein-based cytotoxicity assays such as lactate dehydrogenase (LDH) and tumor necrosis factor-alpha (TNF-α) are commonly used in cytotoxic evaluation of nanoparticles (NPs) despite numerous reports on possible interactions with protein markers in these assays that can confound the results obtained. In this study, conventional cytotoxicity assays where assay markers may (LDH and TNF- α) or may not (PicoGreen and WST-8) come into contact with NPs were used to evaluate the cytotoxicity of NPs. The findings revealed selective interactions between negatively charged protein assay markers (LDH and TNF- α) and positively charged ZnO NPs under abiotic conditions. The adsorption and interaction with these protein assay markers were strongly influenced by surface charge, concentration, and specific surface area of the NPs, thereby resulting in less than accurate cytotoxic measurements, as observed from actual cell viability measurements. An improved protocol for LDH assay was, therefore, proposed and validated by eliminating any effects associated with protein–particle interactions. In view of this, additional measures and precautions should be taken when evaluating cytotoxicity of NPs with standard protein-based assays, particularly when they are of opposite charges.

  10. Nanoparticle-assay marker interaction: effects on nanotoxicity assessment

    Science.gov (United States)

    Zhao, Xinxin; Xiong, Sijing; Huang, Liwen Charlotte; Ng, Kee Woei; Loo, Say Chye Joachim

    2015-01-01

    Protein-based cytotoxicity assays such as lactate dehydrogenase (LDH) and tumor necrosis factor-alpha (TNF-α) are commonly used in cytotoxic evaluation of nanoparticles (NPs) despite numerous reports on possible interactions with protein markers in these assays that can confound the results obtained. In this study, conventional cytotoxicity assays where assay markers may (LDH and TNF- α) or may not (PicoGreen and WST-8) come into contact with NPs were used to evaluate the cytotoxicity of NPs. The findings revealed selective interactions between negatively charged protein assay markers (LDH and TNF- α) and positively charged ZnO NPs under abiotic conditions. The adsorption and interaction with these protein assay markers were strongly influenced by surface charge, concentration, and specific surface area of the NPs, thereby resulting in less than accurate cytotoxic measurements, as observed from actual cell viability measurements. An improved protocol for LDH assay was, therefore, proposed and validated by eliminating any effects associated with protein-particle interactions. In view of this, additional measures and precautions should be taken when evaluating cytotoxicity of NPs with standard protein-based assays, particularly when they are of opposite charges.

  11. Fluorescence-PCR Assays and Isolation of Luminescent Bacterial Clones Using an Automated Plate Reader

    Science.gov (United States)

    Crowley, Thomas E.

    2011-01-01

    The genes responsible for luminescence in various species of the marine microorganism "Photobacterium", have been used for many years as a tool by researchers and instructors. In particular, the "lux" operon of "Photobacterium fischeri" has been used by many instructors to teach recombinant DNA techniques. Two methods using an automated plate…

  12. A simple and rapid plate assay for screening of inulin- degrading ...

    African Journals Online (AJOL)

    Jane

    2011-08-22

    -Nelson method, while their hydrolysis ... solubility at such low temperature, while the products of ... medium and inulinolytic activity was detected by the ... tion was subtracted with thermally inactivated crude enzyme as the.

  13. Assaying Cellular Viability Using the Neutral Red Uptake Assay.

    Science.gov (United States)

    Ates, Gamze; Vanhaecke, Tamara; Rogiers, Vera; Rodrigues, Robim M

    2017-01-01

    The neutral red uptake assay is a cell viability assay that allows in vitro quantification of xenobiotic-induced cytotoxicity. The assay relies on the ability of living cells to incorporate and bind neutral red, a weak cationic dye, in lysosomes. As such, cytotoxicity is expressed as a concentration-dependent reduction of the uptake of neutral red after exposure to the xenobiotic under investigation. The neutral red uptake assay is mainly used for hazard assessment in in vitro toxicology applications. This method has also been introduced in regulatory recommendations as part of 3T3-NRU-phototoxicity-assay, which was regulatory accepted in all EU member states in 2000 and in the OECD member states in 2004 as a test guideline (TG 432). The present protocol describes the neutral red uptake assay using the human hepatoma cell line HepG2, which is often employed as an alternative in vitro model for human hepatocytes. As an example, the cytotoxicity of acetaminophen and acetyl salicylic acid is assessed.

  14. A comparative study on the customized design of mandibular reconstruction plates using finite element method

    Directory of Open Access Journals (Sweden)

    Abdulrahman Al-Ahmari

    2015-07-01

    Full Text Available Mandible defects and its deformities are serious complications and its precise reconstruction is one of the most challenging tasks in oral maxillofacial surgery. The commercially available standard mandible implants are manually bended before surgery to custom fit the patient’s jaw. A slight mismatch in the plate and bone alignment may result in the implant failure. However, with the integration of computer-aided design, rapid prototyping, and advanced imaging systems (computed tomography or magnetic resonance imaging, it is possible to produce a customized mandible implant that can precisely fit the patient’s jaw. The aim of this article is to compare a new design of customized mandible implant (sinewave plate and compare it with the commonly used straight implant design. The finite element–simulated results reveal that the commonly used straight reconstruction plates are more prone to loosening of the screws due to its higher strain concentration on the screw hole when compared to newly designed sinewave reconstruction plate. Moreover, the straight plate is more sensitive to the chewing load variations and develops almost 20% increase in the stresses when compared to sinewave plate. The study reveals that the sinewave reconstruction plate can significantly enhance the stability and safety of the mandible implant.

  15. A New Type of Inscribed Copper Plate from Indus Valley (Harappan Civilisation

    Directory of Open Access Journals (Sweden)

    Vasant Shinde

    2014-10-01

    Full Text Available A group of nine Indus Valley copper plates (c. 2600–2000 BC, discovered from private collections in Pakistan, appear to be of an important type not previously described. The plates are significantly larger and more robust than those comprising the corpus of known copper plates or tablets, and most significantly differ in being inscribed with mirrored characters. One of the plates bears 34 characters, which is the longest known single Indus script inscription. Examination of the plates with x-ray fluorescence (XRF spectrophotometry indicates metal compositions, including arsenical copper, consistent with Indus Valley technology. Microscopy of the metal surface and internal structure reveals detail such as pitting, microcrystalline structure, and corrosion, consistent with ancient cast copper artifacts. Given the relative fineness of the engraving, it is hypothesised that the copper plates were not used as seals, but have characteristics consistent with use in copper plate printing. As such, it is possible that these copper plates are by far the earliest known printing devices, being at least 4000 years old.

  16. Elastoplastic Stability and Failure Analysis of FGM Plate with Temperature Dependent Material Properties under Thermomechanical Loading

    Directory of Open Access Journals (Sweden)

    Kanishk Sharma

    Full Text Available Abstract The present paper explores the stability and failure response of elastoplastic Ni/Al2O3 functionally graded plate under thermomechanical load using non-linear finite element formulation based on first-order shear deformation theory and von-Karman’s nonlinear kinematics. The temperature dependent thermoelastic material properties of FGM plate are varied in the thickness direction by controlling the volume fraction of the constituent materials (i.e., ceramic and metal with a power law, and Mori-Tanaka homogenization scheme is applied to evaluate the properties at a particular thickness coordinate of FGM plate. The elastoplastic behavior of FGM plate is assumed to follow J2-plasticity with isotropic hardening, wherein the ceramic phase is considered to be elastic whereas the metal is assumed to be elastic-plastic in accordance with the Tamura-Tomota-Ozawa model. Numerical studies are conducted to examine the effects of material and geometrical parameters, viz. material in-homogeneity, slenderness and aspect ratios on the elastoplastic bucking and postbuckling behavior and the failure response of FGM plate. It is revealed that material gradation affects the stability and failure behavior of FGM plate considerably. Furthermore, it is also concluded that FGM plate with elastic material properties exhibits only stable equilibrium path, whereas the elastoplastic FGM plate shows destabilizing response after the ultimate failure point.

  17. Rapidly Moving Divertor Plates In A Tokamak

    International Nuclear Information System (INIS)

    Zweben, S.

    2011-01-01

    It may be possible to replace conventional actively cooled tokamak divertor plates with a set of rapidly moving, passively cooled divertor plates on rails. These plates would absorb the plasma heat flux with their thermal inertia for ∼10-30 sec, and would then be removed from the vessel for processing. When outside the tokamak, these plates could be cooled, cleaned, recoated, inspected, and then returned to the vessel in an automated loop. This scheme could provide nearoptimal divertor surfaces at all times, and avoid the need to stop machine operation for repair of damaged or eroded plates. We describe various possible divertor plate designs and access geometries, and discuss an initial design for a movable and removable divertor module for NSTX-U.

  18. Elasto-plastic deformation and plate weakening due to normal faulting in the subducting plate along the Mariana Trench

    Science.gov (United States)

    Zhou, Zhiyuan; Lin, Jian

    2018-06-01

    We investigated variations in the elasto-plastic deformation of the subducting plate along the Mariana Trench through an analysis of flexural bending and normal fault characteristics together with geodynamic modeling. Most normal faults were initiated at the outer-rise region and grew toward the trench axis with strikes mostly subparallel to the local trench axis. The average trench relief and maximum fault throws were measured to be significantly greater in the southern region (5 km and 320 m, respectively) than the northern and central regions (2 km and 200 m). The subducting plate was modeled as an elasto-plastic slab subjected to tectonic loading at the trench axis. The calculated strain rates and velocities revealed an array of normal fault-like shear zones in the upper plate, resulting in significant faulting-induced reduction in the deviatoric stresses. We then inverted for solutions that best fit the observed flexural bending and normal faulting characteristics, revealing normal fault penetration to depths of 21, 20, and 32 km beneath the seafloor for the northern, central, and southern regions, respectively, which is consistent with the observed depths of the relocated normal faulting earthquakes in the central Mariana Trench. The calculated deeper normal faults of the southern region might lead to about twice as much water being carried into the mantle per unit trench length than the northern and central regions. We further calculated that normal faulting has reduced the effective elastic plate thickness Te by up to 52% locally in the southern region and 33% in both the northern and central regions. The best-fitting solutions revealed a greater apparent angle of the pulling force in the southern region (51-64°) than in the northern (22-35°) and central (20-34°) regions, which correlates with a general southward increase in the seismically-determined dip angle of the subducting slab along the Mariana Trench.

  19. Sigmoid plate dehiscence: Congenital or acquired condition?

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Zhaohui, E-mail: lzhtrhos@163.com [Capital Medical University, Beijing Tongren Hospital, No 1 Dong Jiao Min Street, Dongcheng District, Beijing 100730 (China); Li, Jing, E-mail: lijingxbh@yahoo.com.cn [Capital Medical University, Beijing Tongren Hospital, No 1 Dong Jiao Min Street, Dongcheng District, Beijing 100730 (China); Zhao, Pengfei, E-mail: zhaopengf05@163.com [Capital Medical University, Beijing Friendship Hospital, No 95 Yongan Road, Xicheng District, Beijing 100050 (China); Lv, Han, E-mail: chrislvhan@126.com [Capital Medical University, Beijing Friendship Hospital, No 95 Yongan Road, Xicheng District, Beijing 100050 (China); Dong, Cheng, E-mail: derc007@sina.com [Capital Medical University, Beijing Friendship Hospital, No 95 Yongan Road, Xicheng District, Beijing 100050 (China); Liu, Wenjuan, E-mail: wenjuanliu@163.com [Jining No. 1 People' s Hospital, No. 6 Health Street, Jining 272100 (China); Wang, Zhenchang, E-mail: cjr.wzhch@vip.163.com [Capital Medical University, Beijing Friendship Hospital, No 95 Yongan Road, Xicheng District, Beijing 100050 (China)

    2015-05-15

    Highlights: • CT with multiplanar reformations can accurately display the sigmoid platet dehiscence. • The prevalence of sigmoid plate dehiscence was no significant difference among different age groups. • The size of sigmoid plate bony defects were not statistically different among different age groups. • The sigmoid plate dehiscence is more commonly a congenital than an acquired condition. - Abstract: Background and purpose: The imaging features of sigmoid plate dehiscence-induced pulsatile tinnitus have been presented. The origin of the sigmoid plate dehiscence, however, remains unclear. Our aim was to assess the prevalence and extent of sigmoid plate dehiscence on computed tomography (CT) images in multiple age groups to determine whether this condition is more likely to be congenital or acquired. Materials and methods: We retrospectively reviewed contrast-enhanced CT images of sigmoid plates of temporal bones in 504 patients. Each temporal bone was characterized as normal or dehiscent. Patients were then subcategorized into four age groups, and the prevalence and extent of dehiscent sigmoid plates in each group were calculated and compared. Results: Overall, 80 patients had sigmoid plate dehiscence, nine of whom had it bilaterally. In successively older age groups, the prevalences of sigmoid plate dehiscence were 18.9%, 20.1%, 14.5%, and 12.7%, respectively. Respective average anteroposterior bony defect diameters were 3.7 ± 1.7, 3.0 ± 1.3, 3.1 ± 1.5, and 3.0 ± 1.1 mm. Respective average vertical bony defect diameters were 3.6 ± 2.3, 2.6 ± 1.2, 3.2 ± 1.5, and 3.0 ± 1.7 mm. The prevalence and extent of sigmoid plate dehiscence were not statistically different among the four age groups. Conclusions: The similar radiologic prevalence and extent of dehiscent sigmoid plates among the age groups suggest that the dehiscence is more commonly a congenital than an acquired condition.

  20. Strain resolving method of composite plane plates

    Directory of Open Access Journals (Sweden)

    Ion FUIOREA

    2011-06-01

    Full Text Available The paper deals with the extension of isotropic plates problem to the case of composite plates. In order to perform it, the Kirchhoff-Love hypotheses were “softened” by some additional ones. Considering the constitutive laws for composite materials the stress functions were eliminated by using Cauchy equations. As a result a partial derivative equation in displacements was obtained. Finally the boundary condition formulation was extended for the case of complex composite plates.

  1. Empirical Study of Car License Plates Recognition

    OpenAIRE

    Dina, Nasa Zata; Dailey, Matthew

    2015-01-01

    The number of vehicles on the road has increased drastically in recent years. The license plate is an identity card for a vehicle. It can map to the owner and further information about vehicle. License plate information is useful to help traffic management systems. For example, traffic management systems can check for vehicles moving at speeds not permitted by law and can also be installed in parking areas to se-cure the entrance or exit way for vehicles. License plate recognition algorithms ...

  2. 431K/CD vehicle number plates

    CERN Multimedia

    GS Department

    2009-01-01

    The Green Plates Service, which is responsible for issuing the 431K/CD vehicle number plates , wishes to apologise for the delay in processing applications over the past weeks. The delay is outside the Service’s control, as it is due to the recent introduction of new rules governing the vehicle registration process in France. Normal service will be resumed as soon as possible. Thank you for your understanding. GS-SEM-LS – Green Plates Service

  3. Replacement divider plate performance under LOCA loading

    International Nuclear Information System (INIS)

    Huynk, H.M.; MClellan, G.H.; Schneider, W.G.

    1997-01-01

    A primary divider plate in a nuclear steam generator is required to perform its partitioning function with a minimum of cross leakage, without degradation in operating performance and without loss of structural integrity resulting from normal and accident loading. The design of the replacement divider plate for normal operating conditions is discussed in some detail in reference 1 and 2. This paper describes the structural response of the replacement divider plate to the severe loading resulting from a burst primary pipe. The loads for which the divider plate structural performance must be evaluated are mild to severe differential pressure transients resulting from several postulated sizes and types of pipe break scenarios. In the unlikely event of a severe Loss of Coolant Accident (LOCA) the divider plate or parts thereof must not exit the steam generator nor completely block the outlet nozzle. For the milder LOCA loads, the integrity of the divider plate and seat bars must be maintained. Analysis for the milder LOCA loads was carried out employing a conservative approach which ignores the actual interaction between the structure and the primary fluid. For these load cases it was shown that the divider plate does not become disengaged from the seat bars. For the more severe pipe breaks, the thermal-hydraulic analysis was coupled iteratively with the structural analysis, thereby taking into account divider plate deformation, in order to obtain a better prediction of the behaviour of the divider plate. In this manner substantial reduction in divider plate response to the more severe LOCA loading was achieved. It has been shown that, for the case of a postulated large LOCA (100% reactor inlet header), the disengagement of the divider plate from the seat bars resulted in an opening smaller than 1% of the divider plate area. (author)

  4. Sigmoid plate dehiscence: Congenital or acquired condition?

    International Nuclear Information System (INIS)

    Liu, Zhaohui; Li, Jing; Zhao, Pengfei; Lv, Han; Dong, Cheng; Liu, Wenjuan; Wang, Zhenchang

    2015-01-01

    Highlights: • CT with multiplanar reformations can accurately display the sigmoid platet dehiscence. • The prevalence of sigmoid plate dehiscence was no significant difference among different age groups. • The size of sigmoid plate bony defects were not statistically different among different age groups. • The sigmoid plate dehiscence is more commonly a congenital than an acquired condition. - Abstract: Background and purpose: The imaging features of sigmoid plate dehiscence-induced pulsatile tinnitus have been presented. The origin of the sigmoid plate dehiscence, however, remains unclear. Our aim was to assess the prevalence and extent of sigmoid plate dehiscence on computed tomography (CT) images in multiple age groups to determine whether this condition is more likely to be congenital or acquired. Materials and methods: We retrospectively reviewed contrast-enhanced CT images of sigmoid plates of temporal bones in 504 patients. Each temporal bone was characterized as normal or dehiscent. Patients were then subcategorized into four age groups, and the prevalence and extent of dehiscent sigmoid plates in each group were calculated and compared. Results: Overall, 80 patients had sigmoid plate dehiscence, nine of whom had it bilaterally. In successively older age groups, the prevalences of sigmoid plate dehiscence were 18.9%, 20.1%, 14.5%, and 12.7%, respectively. Respective average anteroposterior bony defect diameters were 3.7 ± 1.7, 3.0 ± 1.3, 3.1 ± 1.5, and 3.0 ± 1.1 mm. Respective average vertical bony defect diameters were 3.6 ± 2.3, 2.6 ± 1.2, 3.2 ± 1.5, and 3.0 ± 1.7 mm. The prevalence and extent of sigmoid plate dehiscence were not statistically different among the four age groups. Conclusions: The similar radiologic prevalence and extent of dehiscent sigmoid plates among the age groups suggest that the dehiscence is more commonly a congenital than an acquired condition

  5. Evaluation of korzincalloy prepared by Hohman Plating

    Energy Technology Data Exchange (ETDEWEB)

    Korinko, P. S. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Hollingshad, A. N. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2017-07-17

    A commercial vendor, Hohman Plating performed contract engineering work to determine the feasibility of producing pin hole free KorZincAlloy bronze material used for zinc gettering. Samples were tested for Sn plating thickness, heat treatability, and chemistry prior to being subjected to a standardized zinc exposure. The samples absorbed zinc and were examined using visual and scanning electron microscopy. Hohman Plating successfully produced KZA that met the target composition, was pin hole free, and was an effective zinc getter.

  6. Plasma myelin basic protein assay using Gilford enzyme immunoassay cuvettes.

    Science.gov (United States)

    Groome, N P

    1981-10-01

    The assay of myelin basic protein in body fluids has potential clinical importance as a routine indicator of demyelination. Preliminary details of a competitive enzyme immunoassay for this protein have previously been published by the author (Groome, N. P. (1980) J. Neurochem. 35, 1409-1417). The present paper now describes the adaptation of this assay for use on human plasma and various aspects of routine data processing. A commercially available cuvette system was found to have advantages over microtitre plates but required a permuted arrangement of sample replicates for consistent results. For dose interpolation, the standard curve could be fitted to a three parameter non-linear equation by regression analysis or linearised by the logit/log transformation.

  7. Automated 5 ' nuclease PCR assay for identification of Salmonella enterica

    DEFF Research Database (Denmark)

    Hoorfar, Jeffrey; Ahrens, Peter; Rådström, P.

    2000-01-01

    -point fluorescence (FAM) signals for the samples and positive control (TET) signals (relative sensitivity [Delta Rn], >0.6). The diagnostic specificity of the method was assessed using 120 non-Salmonella strains, which all resulted in negative FAM signals (Delta Rn, less than or equal to 0.5). All 100 rough...... Salmonella strains tested resulted in positive FAM and TET signals. In addition, it was found that the complete PCR mixture, predispensed in microwell plates, could be stored for up to 3 months at -20 degrees C, Thus, the diagnostic TaqMan assay developed can be a useful and simple alternative method......A simple and ready-to-go test based on a 5' nuclease (TaqMan) PCR technique was developed for identification of presumptive Salmonella enterica isolates. The results were compared with those of conventional methods. The TaqMan assay was evaluated for its ability to accurately detect 210 S. enterica...

  8. 3-D Simulation of Tectonic Evolution in Mariana with a Coupled Model of Plate Subduction and Back-Arc Spreading

    Science.gov (United States)

    Hashima, A.; Matsu'Ura, M.

    2006-12-01

    We obtained the expressions for internal deformation fields due to a moment tensor in an elastic-viscoelastic layered holf-space. This unified formulation of internal deformation fields for shear faulting and crack opening enabled us to deal with the problem of tectonic deformation at a composite type of plate boundary zones. The tectonic deformation can be ascribed to mechanical interaction at plate boundaries, which make a closed circuit with the mode of relative plate motion changing from divergence to convergence through transcurrent motion. One of the rational ways to represent mechanical interaction at plate boundaries is specifying the increase rates of normal or tangential displacement discontinuity across plate interfaces. On the basis of such a basic idea we developed a 3-D simulation model for the nonlinear, coupled system of plate subduction and back-arc spreading in Mariana. Through numerical simulations we revealed the evolution process of back-arc spreading. At the first stage, steady plate subduction (shear faulting at a plate interface) gradually forms tensile stress fields in the back-arc region of the overriding plate. When the accumulated tensile stress reaches a critical level, back-arc spreading (crack opening) starts at a structurally weak portion of the overriding plate. The horizontal motion of the frontal part of the overriding plate due to back-arc spreading pushes out the plate boundary toward the oceanic plate. In steady-state plate subduction the shear stress acting on a plate interface must balance with the maximum frictional resistance (shear strength) of the plate interface. Therefore, the increase of shear stress at the plate interface leads to the increase of slip rate at the plate interface. The local increase of slip rate at the plate interface produces the additional tensile stress in the back-arc region. The increased tensile stress must be canceled out by the additional crack opening. Such a feedback mechanism between plate

  9. Solution assay instrument operations manual

    International Nuclear Information System (INIS)

    Li, T.K.; Marks, T.; Parker, J.L.

    1983-09-01

    An at-line solution assay instrument (SAI) has been developed and installed in a plutonium purification and americium recovery process area in the Los Alamos Plutonium Processing Facility. The instrument was designed for accurate, timely, and simultaneous nondestructive analysis of plutonium and americium in process solutions that have a wide range of concentrations and americium/plutonium ratios and for routine operation by process technicians who lack instrumentation background. The SAI, based on transmission-corrected, high-resolution gamma-ray spectroscopy, has two measurement stations attached to a single multichannel analyzer/computer system. To ensure the quality of assay results, the SAI has an internal measurement control program, which requires daily and weekly check runs and monitors key aspects of all assay runs. For a 25-ml sample, the assay precision is 5 g/l within a 2000-s count time

  10. Locking screw-plate interface stability in carbon-fibre reinforced polyetheretherketone proximal humerus plates.

    Science.gov (United States)

    Hak, David J; Fader, Ryan; Baldini, Todd; Chadayammuri, Vivek B S

    2017-09-01

    Carbon-fibre reinforced polyetheretherketone (CFR-PEEK) plates have recently been introduced for proximal humerus fracture treatment. The purpose of this study was to compare the locking screw-plate interface stability in CFR-PEEK versus stainless steel (SS) proximal humerus plates. Locking screw mechanical stability was evaluated independently in proximal and shaft plate holes. Stiffness and load to failure were tested for three conditions: (1) on-axis locking screw insertion in CFR-PEEK versus SS plates, (2) on-axis locking screw insertion, removal, and reinsertion in CFR-PEEK plates, and (3) 10-degree off-axis locking screw insertion in CFR-PEEK plates. Cantilever bending at a rate of 1 mm/minute was produced by an Instron machine and load-displacement data recorded. Shaft locking screw load to failure was significantly greater in CFR-PEEK plates compared to SS plates (746.4 ± 89.7 N versus 596.5 ± 32.6 N, p PEEK plates (p PEEK plates. The mechanical stability of locking screws in CFR-PEEK plates is comparable or superior to locking screws in SS plates.

  11. Fuel cell cooler-humidifier plate

    Science.gov (United States)

    Vitale, Nicholas G.; Jones, Daniel O.

    2000-01-01

    A cooler-humidifier plate for use in a proton exchange membrane (PEM) fuel cell stack assembly is provided. The cooler-humidifier plate combines functions of cooling and humidification within the fuel cell stack assembly, thereby providing a more compact structure, simpler manifolding, and reduced reject heat from the fuel cell. Coolant on the cooler side of the plate removes heat generated within the fuel cell assembly. Heat is also removed by the humidifier side of the plate for use in evaporating the humidification water. On the humidifier side of the plate, evaporating water humidifies reactant gas flowing over a moistened wick. After exiting the humidifier side of the plate, humidified reactant gas provides needed moisture to the proton exchange membranes used in the fuel cell stack assembly. The invention also provides a fuel cell plate that maximizes structural support within the fuel cell by ensuring that the ribs that form the boundaries of channels on one side of the plate have ends at locations that substantially correspond to the locations of ribs on the opposite side of the plate.

  12. Maximizing band gaps in plate structures

    DEFF Research Database (Denmark)

    Halkjær, Søren; Sigmund, Ole; Jensen, Jakob Søndergaard

    2006-01-01

    periodic plate using Bloch theory, which conveniently reduces the maximization problem to that of a single base cell. Secondly, we construct a finite periodic plate using a number of the optimized base cells in a postprocessed version. The dynamic properties of the finite plate are investigated......Band gaps, i.e., frequency ranges in which waves cannot propagate, can be found in elastic structures for which there is a certain periodic modulation of the material properties or structure. In this paper, we maximize the band gap size for bending waves in a Mindlin plate. We analyze an infinite...... theoretically and experimentally and the issue of finite size effects is addressed....

  13. Metallized xerographic plates and their preparation

    International Nuclear Information System (INIS)

    1976-01-01

    Xerographic plates, particularly xeroradiographic plates, suitable for soft or hard x-ray exposure require margins or edges suitable for handling and mounting. In order to fabricate such plates, it is convenient to apply a peripheral mask over the substrate during application of at least the photoconductive layer. Use of masks, however, tend to promote peripheral plate irregularities usually because of scratches or other uneveness at or under the margin of the applied mask. Such imperfections spawn corresponding electrical field irregularities which can now be minimized or avoided altogether by grounding the margins with metal overcoats

  14. Bipolar plates for PEM fuel cells

    Science.gov (United States)

    Middelman, E.; Kout, W.; Vogelaar, B.; Lenssen, J.; de Waal, E.

    The bipolar plates are in weight and volume the major part of the PEM fuel cell stack, and are also a significant contributor to the stack costs. The bipolar plate is therefore a key component if power density has to increase and costs must come down. Three cell plate technologies are expected to reach targeted cost price levels, all having specific advantages and drawbacks. NedStack has developed a conductive composite materials and a production process for fuel cell plates (bipolar and mono-polar). The material has a high electric and thermal conductivity, and can be processed into bipolar plates by a proprietary molding process. Process cycle time has been reduced to less than 10 s, making the material and process suitable for economical mass production. Other development work to increase material efficiency resulted in thin bipolar plates with integrated cooling channels, and integrated seals, and in two-component bipolar plates. Total thickness of the bipolar plates is now less than 3 mm, and will be reduced to 2 mm in the near future. With these thin integrated plates it is possible to increase power density up to 2 kW/l and 2 kW/kg, while at the same time reducing cost by integrating other functions and less material use.

  15. Line Heat-Source Guarded Hot Plate

    Data.gov (United States)

    Federal Laboratory Consortium — Description:The 1-meter guarded hot-plate apparatus measures thermal conductivity of building insulation. This facility provides for absolute measurement of thermal...

  16. No quantum friction between uniformly moving plates

    Energy Technology Data Exchange (ETDEWEB)

    Philbin, T G; Leonhardt, U [School of Physics and Astronomy, University of St Andrews, North Haugh St Andrews, Fife KY16 9SS, Scotland (United Kingdom)], E-mail: tgp3@st-andrews.ac.uk

    2009-03-15

    The Casimir forces between two plates moving parallel to each other at arbitrary constant speed are found by calculating the vacuum electromagnetic stress tensor. The perpendicular force between the plates is modified by the motion but there is no lateral force on the plates. Electromagnetic vacuum fluctuations do not therefore give rise to 'quantum friction' in this case, contrary to previous assertions. The result shows that the Casimir-Polder force on a particle moving at constant speed parallel to a plate also has no lateral component.

  17. No quantum friction between uniformly moving plates

    International Nuclear Information System (INIS)

    Philbin, T G; Leonhardt, U

    2009-01-01

    The Casimir forces between two plates moving parallel to each other at arbitrary constant speed are found by calculating the vacuum electromagnetic stress tensor. The perpendicular force between the plates is modified by the motion but there is no lateral force on the plates. Electromagnetic vacuum fluctuations do not therefore give rise to 'quantum friction' in this case, contrary to previous assertions. The result shows that the Casimir-Polder force on a particle moving at constant speed parallel to a plate also has no lateral component.

  18. Peptide Binding to HLA Class I Molecules: Homogenous, High-Throughput Screening, and Affinity Assays

    DEFF Research Database (Denmark)

    Harndahl, Mikkel; Justesen, Sune Frederik Lamdahl; Lamberth, Kasper

    2009-01-01

    , better signal-to-background ratios, and a higher capacity. They also describe an efficient approach to screen peptides for binding to HLA molecules. For the occasional user, this will serve as a robust, simple peptide-HLA binding assay. For the more dedicated user, it can easily be performed in a high-throughput...... the luminescent oxygen channeling immunoassay technology (abbreviated LOCI and commercialized as AlphaScreen (TM)). Compared with an enzyme-linked immunosorbent assay-based peptide-HLA class I binding assay, the LOCI assay yields virtually identical affinity measurements, although having a broader dynamic range...... screening mode using standard liquid handling robotics and 384-well plates. We have successfully applied this assay to more than 60 different HLA molecules, leading to more than 2 million measurements. (Journal of Biomolecular Screening 2009: 173-180)...

  19. Radioligand assay in reproductive biology

    International Nuclear Information System (INIS)

    Korenman, S.G.; Sherman, B.M.

    1975-01-01

    Radioligand assays have been developed for the principal reproductive steroids and peptide hormones. Specific binding reagents have included antibodies, plasma binders, and intracellular receptors. In each assay, problems of specificity, sensitivity, and nonspecific inhibitors were encountered. Many features of the endocrine physiology in childhood, during puberty, and in adulthood have been characterized. Hormonal evaluations of endocrine disorders of reproduction are characterized on the basis of their characteristic pathophysiologic alterations. (U.S.)

  20. Active constrained layer damping of geometrically nonlinear vibrations of functionally graded plates using piezoelectric fiber-reinforced composites

    International Nuclear Information System (INIS)

    Panda, Satyajit; Ray, M C

    2008-01-01

    In this paper, a geometrically nonlinear dynamic analysis has been presented for functionally graded (FG) plates integrated with a patch of active constrained layer damping (ACLD) treatment and subjected to a temperature field. The constraining layer of the ACLD treatment is considered to be made of the piezoelectric fiber-reinforced composite (PFRC) material. The temperature field is assumed to be spatially uniform over the substrate plate surfaces and varied through the thickness of the host FG plates. The temperature-dependent material properties of the FG substrate plates are assumed to be graded in the thickness direction of the plates according to a power-law distribution while the Poisson's ratio is assumed to be a constant over the domain of the plate. The constrained viscoelastic layer of the ACLD treatment is modeled using the Golla–Hughes–McTavish (GHM) method. Based on the first-order shear deformation theory, a three-dimensional finite element model has been developed to model the open-loop and closed-loop nonlinear dynamics of the overall FG substrate plates under the thermal environment. The analysis suggests the potential use of the ACLD treatment with its constraining layer made of the PFRC material for active control of geometrically nonlinear vibrations of FG plates in the absence or the presence of the temperature gradient across the thickness of the plates. It is found that the ACLD treatment is more effective in controlling the geometrically nonlinear vibrations of FG plates than in controlling their linear vibrations. The analysis also reveals that the ACLD patch is more effective for controlling the nonlinear vibrations of FG plates when it is attached to the softest surface of the FG plates than when it is bonded to the stiffest surface of the plates. The effect of piezoelectric fiber orientation in the active constraining PFRC layer on the damping characteristics of the overall FG plates is also discussed

  1. Active constrained layer damping of geometrically nonlinear vibrations of functionally graded plates using piezoelectric fiber-reinforced composites

    Science.gov (United States)

    Panda, Satyajit; Ray, M. C.

    2008-04-01

    In this paper, a geometrically nonlinear dynamic analysis has been presented for functionally graded (FG) plates integrated with a patch of active constrained layer damping (ACLD) treatment and subjected to a temperature field. The constraining layer of the ACLD treatment is considered to be made of the piezoelectric fiber-reinforced composite (PFRC) material. The temperature field is assumed to be spatially uniform over the substrate plate surfaces and varied through the thickness of the host FG plates. The temperature-dependent material properties of the FG substrate plates are assumed to be graded in the thickness direction of the plates according to a power-law distribution while the Poisson's ratio is assumed to be a constant over the domain of the plate. The constrained viscoelastic layer of the ACLD treatment is modeled using the Golla-Hughes-McTavish (GHM) method. Based on the first-order shear deformation theory, a three-dimensional finite element model has been developed to model the open-loop and closed-loop nonlinear dynamics of the overall FG substrate plates under the thermal environment. The analysis suggests the potential use of the ACLD treatment with its constraining layer made of the PFRC material for active control of geometrically nonlinear vibrations of FG plates in the absence or the presence of the temperature gradient across the thickness of the plates. It is found that the ACLD treatment is more effective in controlling the geometrically nonlinear vibrations of FG plates than in controlling their linear vibrations. The analysis also reveals that the ACLD patch is more effective for controlling the nonlinear vibrations of FG plates when it is attached to the softest surface of the FG plates than when it is bonded to the stiffest surface of the plates. The effect of piezoelectric fiber orientation in the active constraining PFRC layer on the damping characteristics of the overall FG plates is also discussed.

  2. Divider plate leak inspections using an acoustic tool 'ALIS'

    International Nuclear Information System (INIS)

    Kowalewski, J.; Colbert, M.

    1998-01-01

    Divider plate leakage in the primary head of steam generators may be a contributor to high Reactor Inlet Header Temperature (RIHT) which can have a negative effect on the operation of a plant. A method to provide quick information of divider plate leakage can be very useful in helping operators to make timely and cost effective repair and maintenance decisions. As part of a CANDU Owners Group (COG) funded program, a novel acoustic tool for performing inspections in empty steam generators during a shutdown has been developed and successfully demonstrated by Ontario Hydro Technologies (OHT). The basic principle of this technique utilizes sound transmission through the leakage paths of a divider plate assembly for creating a graphic image which reveals the locations and relative sizes of leaks. This is accomplished by injecting one side of the primary head with a series of airborne, ultrasonic sound bursts while scanning the divider plates from the opposite side and collecting transmitted sound pressure levels. Once a scan is completed the results are processed and displayed graphically as a color mapping of the entire divider plate assembly showing an image of the leakage locations and relative severity. The special transducers of the acoustic leak inspection system (ALIS) are installed through the manways of the steam generator at both sides of the primary head. These are clamped to the tube sheet and operated from a convenient remote location within the containment building. This process minimizes radiation exposure of the workers, as well, it reduces risk of contamination of the ALIS equipment. The current version of this tool performs the scans by remote manual control. (author)

  3. Assaying the reporter gene chloramphenicol acetyltransferase

    International Nuclear Information System (INIS)

    Crabb, D.W.; Minth, C.D.; Dixon, J.E.

    1989-01-01

    These experiments document the presence of enzymatic activities in extracts of commonly used cell lines which interfere with the determination of CAT activity. We suspect that the deacetylase activity is the most important, as the extract of the H4IIE C3 cells was capable of completely deacetylating the mono- and diacetylchloramphenicol formed during a 2-hr incubation of CAT with chloramphenicol and acetyl-CoA. The results of the inhibitor experiments are consistent with the presence of proteases which degrade CAT, or a serine carboxylesterase. The interference was also reduced by about half by EDTA; a metalloenzyme (either a protease or esterase) may therefore be involved. This interference appears to be a common phenomenon. We have surveyed 23 different cell types for the presence of the interfering activity and found it in 15. The interference was particularly prominent in several neuroendocrine and hepatoma cells. We took advantage of the effect of EDTA and the heat stability of CAT to eliminate the interference. Addition of 5 mM EDTA and a 10-min incubation of the sonicated cell suspension at 60 degrees prior to centrifugation abolished the interference in all cell lines tested. It is important to note that in order to reveal any CAT activity in some of the extracts (e.g., PC-12 or Hep3B), it was necessary to run the CAT assay for 2 hr. The control assays were therefore run almost to completion, and were well beyond the linear range of the assay. Therefore, the small differences which we observed between the heat-treated and control samples in some instances (e.g., rice, corn, or HeLa cells) will be dramatically amplified when the CAT assay is performed under conditions in which only a small percentage of the substrate is converted to product

  4. Application of a Brittle Damage Model to Normal Plate-on-Plate Impact

    National Research Council Canada - National Science Library

    Raftenberg, Martin N

    2005-01-01

    A brittle damage model presented by Grinfeld and Wright of the U.S. Army Research Laboratory was implemented in the LS-DYNA finite element code and applied to the simulation of normal plate-on-plate impact...

  5. Correlation between the genotoxicity endpoints measured by two different genotoxicity assays: comet assay and CBMN assay

    Directory of Open Access Journals (Sweden)

    Carina Ladeira

    2015-06-01

    The results concerning of positive findings by micronuclei and non significant ones by comet assay, are corroborated by Deng et al. (2005 study performed in workers occupationally exposed to methotrexate, also a cytostatic drug. According to Cavallo et al. (2009, the comet assay seems to be more suitable for the prompt evaluation of the genotoxic effects, for instance, of polycyclic aromatic hydrocarbons mixtures containing volatile substances, whereas the micronucleus test seems more appropriate to evaluate the effects of exposure to antineoplastic agents. However, there are studies that observed an increase in both the comet assay and the micronucleus test in nurses handling antineoplastic drugs, although statistical significance was only seen in the comet assay, quite the opposite of our results (Maluf & Erdtmann, 2000; Laffon et al. 2005.

  6. Improved assay for measuring heparin binding to bull sperm

    International Nuclear Information System (INIS)

    Miller, D.J.; Ax, R.L.

    1988-01-01

    The binding of heparin to sperm has been used to study capacitation and to rank relative fertility of bulls. Previous binding assays were laborious, used 10 7 sperm per assay point, and required large amounts of radiolabeled heparin. A modified heparin-binding assay is described that used only 5 x 10 4 cells per incubation well and required reduced amounts of [ 3 H] heparin. The assay was performed in 96-well Millititer plates, enabling easy incubation and filtering. Dissociation constants and concentrations of binding sites did not differ if analyzed by Scatchard plots, Woolf plots, or by log-logit transformed weighted nonlinear least squares regression, except in the case of outliers. In such cases, Scatchard analysis was more sensitive to outliers. Nonspecific binding was insignificant using nonlinear logistic fit regression and a proportion graph. The effects were tested of multiple free-thawing of sperm in either a commercial egg yolk extender, 40 mM Tris buffer with 8% glycerol, or 40 mM Tris buffer without glycerol. Freeze-thawing in extender did not affect the dissociation constant or the concentration of binding sites. However, freeze-thawing three times in 40 mM Tris reduced the concentration of binding sites and lowered the dissociation constant (raised the affinity). The inclusion of glycerol in the 40 mM Tris did not significantly affect the estimated dissociation constant or the concentration of binding sites as compared to 40 mM Tris without glycerol

  7. A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids

    Science.gov (United States)

    Zipper, Lauren E.; Aristide, Xavier; Bishop, Dylan P.; Joshi, Ishita; Kharzeev, Julia; Patel, Krishna B.; Santiago, Brianna M.; Joshi, Karan; Dorsinvil, Kahille; Sweet, Robert M.; Soares, Alexei S.

    2014-01-01

    A method is described for using plate lids to reduce evaporation in low-volume vapor-diffusion crystallization experiments. The plate lids contain apertures through which the protein and precipitants were added to different crystallization microplates (the reservoir was filled before fitting the lids). Plate lids were designed for each of these commonly used crystallization microplates. This system minimizes the dehydration of crystallization droplets containing just a few nanolitres of protein and precipitant, and results in more reproducible diffraction from the crystals. For each lid design, changes in the weight of the plates were used to deduce the rate of evaporation under different conditions of temperature, air movement, droplet size and precipitant. For comparison, the state of dehydration was also visually assessed throughout the experiment. Finally, X-ray diffraction methods were used to compare the diffraction of protein crystals that were conventionally prepared against those that were prepared on plates with plate lids. The measurements revealed that the plate lids reduced the rate of evaporation by 63–82%. Crystals grown in 5 nl drops that were set up with plate lids diffracted to higher resolution than similar crystals from drops that were set up without plate lids. The results demonstrate that plate lids can be instrumental for improving few-nanolitre crystallizations. PMID:25484231

  8. Steel plate reinforcement of orthotropic bridge decks

    NARCIS (Netherlands)

    Teixeira de Freitas, S.

    2012-01-01

    The PhD research is focused on the reinforcement of fatigue cracked orthotropic steel bridge decks (OBD) by adding a second steel plate to the existing deck. The main idea is to stiffen the existing deck plate, which will reduce the stresses at the fatigue sensitive details and extend the fatigue

  9. Plastic collapse load of corroded steel plates

    Indian Academy of Sciences (India)

    Keywords. Corroded steel plate; plastic collapse; FEM; rough surface. ... The main aim of present work is to study plastic collapse load of corroded steel plates with irregular surfaces under tension. Non-linear finite element method ... Department of Ocean Engineering, AmirKabir University of Technology, 15914 Tehran, Iran ...

  10. DOES TUBULARIZED INCISED PLATE URETHROPLASTY FIT

    African Journals Online (AJOL)

    posterior or proximal hypospadias. Patient age ranged from 22 months to 9 years (mean 5.4 years). Twenty-two cases were primary, while. 10 cases were repeat cases after failed onlay procedures with preserved urethral plate. Cases with severe chordee necessitating ex- cision of the urethral plate or with a very thin.

  11. Method of nickel-plating large components

    International Nuclear Information System (INIS)

    Wilbuer, K.

    1997-01-01

    The invention concerns a method of nickel-plating components, according to which even large components can be provided with an adequate layer of nickel which is pore- and stress-free and such that water is not lost. According to the invention, the component is heated and, after heating, is pickled, rinsed, scoured, plated in an electrolysis process, and rinsed again. (author)

  12. Improved nickel plating of Inconel X-750

    Science.gov (United States)

    Farmer, M. E.; Feeney, J. E.; Kuster, C. A.

    1969-01-01

    Electroplating technique with acid pickling provides a method of applying nickel plating on Inconel X-750 tubing to serve as a wetting agent during brazing. Low-stress nickel-plating bath contains no organic wetting agents that cause the nickel to blister at high temperatures.

  13. Thermal performance of plate-type loop thermosyphon at sub-atmospheric pressures

    International Nuclear Information System (INIS)

    Tsoi, Vadim; Chang, Shyy Woei; Chiang Kuei Feng; Huang, Chuan Chin

    2011-01-01

    This experimental study examines the thermal performance of a newly devised plate-type two-phase loop thermosyphon with cooling applications to electronic boards of telecommunication systems. The evaporation section is configured as the inter-connected multi channels to emulate the bridging boiling mechanism in pulsating thermosyphon. Two thermosyphon plates using water as the coolant with filling ratios (FR) of 0.22 and 0.32 are tested at sub-atmospheric pressures. The vapor-liquid flow images as well as the thermal resistances and effective spreading thermal conductivities are individually measured for each thermosyphon test plate at various heating powers. The high-speed digital images of the vapor-liquid flow structures reveal the characteristic boiling phenomena and the vapor-liquid circulation in the vertical thermosyphon plate, which assist to explore the thermal physics for this type of loop thermosyphon. The bubble agglomeration and pumping action in the inter-connected boiling channels take place at metastable non-equilibrium conditions, leading to the intermittent slug flows with a pulsation character. Such hybrid loop-pulsating thermosyphon permits the vapor-liquid circulation in the horizontal plate. Thermal resistances and spreading thermal conductivities detected from the present thermosyphon plates; the vapor chamber flat plate heat pipe and the copper plate at free and forced convective cooling conditions with both vertical and horizontal orientations are cross-examined. In most telecommunication systems and units, the electrical boards are vertical so that the thermal performance data on the vertical thermosyphon are most relevant to this particular application. - Highlights: → We examine thermal performances of plate-type loop thermosyphon. → Thermal resistances and spreading conductivities are examined. → Bubble agglomeration in inter-connected boiling channels generates intermittent slug flows with pulsations. → Boiling instability

  14. Isolated tympanic plate fracture frequency and its relationship to mandibular trauma.

    Science.gov (United States)

    Altay, Canan; Erdoğan, Nezahat; Batkı, Ozan; Eren, Erdem; Altay, Sedat; Karasu, Sebnem; Mete, Berna; Uluç, Engin

    2014-11-01

    This study evaluated the prevalence of isolated tympanic fractures and their correlation with mandibular fractures by using maxillofacial computed tomography (CT). We retrospectively evaluated the maxillofacial CT of 1590 patients who presented to our emergency department with maxillofacial trauma between December 2010 and December 2012. Maxillofacial CT was used as the criterion standard for evaluating patients with maxillofacial fractures. The CT images were evaluated by using an electronic picture archiving and communications system and interpreted independently by 2 radiologists. The maxillofacial CT images revealed mandibular fractures in 167 of the patients and isolated tympanic plate fractures in 35 of these 167 patients. Four patients (11%) had a bilateral tympanic plate fracture, and 31 patients (89%) had unilateral tympanic plate fracture. Of all the tympanic plate fractures, 19 (54%) were on the right side and 16 (46%) were on the left side (P > .05). In our results, a significant correlation between the presence of a right-sided tympanic plate fracture and fracture of the ipsilateral condylar process was found (P = .036). However, a statistically significant difference between the presence of a tympanic plate fracture and other mandible fractures, additional soft-tissue findings, or the number of fractures was not determined (P > .05). Sex had no impact on the presence of tympanic plate fracture (P > .05). The frequency of isolated tympanic plate fractures in maxillofacial trauma is low, but it is an important anatomic location. Condyle fractures are significantly associated with isolated tympanic plate fractures. The presence of these injuries should raise suspicion of a concomitant isolated tympanic plate fracture. Copyright © 2014 Canadian Association of Radiologists. Published by Elsevier Inc. All rights reserved.

  15. Immunoradiometric assay for cytomegalovirus-specific IgG antibodies; Assay development and evaluation in blood transfusion practice

    Energy Technology Data Exchange (ETDEWEB)

    Klapper, P.E.; Cleator, G.M.; Prinja-Wolks, D.; Morris, D.J. (Medical School, Manchester (United Kingdom). Department of Medical microbiology, Virology Unit); Morell, G. (Regional Blood Transfusion Centre, manchester (United Kingdom))

    1990-03-01

    An immunoradiometric assay (radio-immunosorbent test; RIST) for the detection of IgG antibodies to human herpesvirus 4 (human cytomegalovirus (CMV)) has been developed. The technique utilizes CMV antigen passively adsorbed to a polyvinyl microtitration plate and a radiolabelled murine monoclonal anti-human IgG antibody to detect binding of human antibody to the 'solid phase' reagent. The assay was optimized, and its specifity confirmed by testing paired acute and convalescent sera from patients with acute CMV or other human herpesvirus infections. To determine the assay's sensitivity 1433 blood donor sera were examined. The RIST was more sensitive than a standard complement fixation (CFT). Use of a monoclonal anti-human IgG antibody in the RIST reduced non-specific binding to the control uninfected cell antigen such that blood donor sera could be tested in the assay using only a CMV antigen without generating an unacceptable false positive rate. (author). 23 refs.; 1 tab.

  16. Direct Competitive Enzyme-Linked Immunosorbent Assay (ELISA).

    Science.gov (United States)

    Kohl, Thomas O; Ascoli, Carl A

    2017-07-05

    The competitive enzyme-linked immunosorbent assay (ELISA) (cELISA; also called an inhibition ELISA) is designed so that purified antigen competes with antigen in the test sample for binding to an antibody that has been immobilized in microtiter plate wells. The same concept works if the immobilized molecule is antigen and the competing molecules are purified labeled antibody versus antibody in a test sample. Direct cELISAs incorporate labeled antigen or antibody, whereas indirect assay configurations use reporter-labeled secondary antibodies. The cELISA is very useful for determining the concentration of small-molecule antigens in complex sample mixtures. In the direct cELISA, antigen-specific capture antibody is adsorbed onto the microtiter plate before incubation with either known standards or unknown test samples. Enzyme-linked antigen (i.e., labeled antigen) is also added, which can bind to the capture antibody only when the antibody's binding site is not occupied by either the antigen standard or antigen in the test samples. Unbound labeled and unlabeled antigens are washed away and substrate is added. The amount of antigen in the standard or the test sample determines the amount of reporter-labeled antigen bound to antibody, yielding a signal that is inversely proportional to antigen concentration within the sample. Thus, the higher the antigen concentration in the test sample, the less labeled antigen is bound to the capture antibody, and hence the weaker is the resultant signal. © 2017 Cold Spring Harbor Laboratory Press.

  17. Is There Really A North American Plate?

    Science.gov (United States)

    Krill, A.

    2011-12-01

    Lithospheric plates are typically identified from earthquake epicenters and evidence such as GPS movements. But no evidence indicates a plate boundary between the North American and South American Plates. Some plate maps show them separated by a transform boundary, but it is only a fracture zone. Other maps show an "undefined plate boundary" or put no boundary between these two plates (check Google images). Early plate maps showed a single large American Plate, quite narrow east of the Caribbean Plate (Le Pichon 1968, Morgan 1968). The North and South American Plates became established by the leading textbook Earth (Press & Siever 1974). On their map, from a Scientific American article by John Dewey (1972), these new plates were separated by an "uncertain plate boundary." The reasons for postulating a North American Plate were probably more psychological than geological. Each of the other continents of the world had its own plate, and North American geologists naturally wanted theirs. Similarly, European geographers used to view Europe as its own continent. A single large plate should again be hypothesized. But the term American Plate would now be ambiguous ("Which plate, North or South?") Perhaps future textbook authors could call it the "Two-American Plate." Textbook authors ultimately decide such global-tectonic matters. I became aware of textbook authors' opinions and influence from my research into the history of Alfred Wegener's continental drift (see Fixists vs. Mobilists by Krill 2011). Leading textbook author Charles Schuchert realized that continental drift would abolish his cherished paleogeographic models of large east-west continents (Eria, Gondwana) and small oceans (Poseiden, Nereis). He and his junior coauthors conspired to keep drift evidence out of their textbooks, from the 1934-editions until the 1969-editions (Physical Geology by Longwell et al. 1969, Historical Geology by Dunbar & Waage 1969). Their textbooks ruled in America. Textbooks

  18. Blast response of corroded steel plates

    Energy Technology Data Exchange (ETDEWEB)

    Eslamimajd, Alireza; RahbarRanji, Ahmad [AmirKabir University of Technology, Tehran (Korea, Republic of)

    2014-05-15

    Numerical results for one- and both-sided corroded steel plates subjected to blast loading are presented. Finite element analysis, with ABAQUS software, is employed to determine the deformation and stress distributions. The results for the case of triangular pulse pressure on un-corroded plates are validated against literature-based data and then, detailed parametric studies are carried-out. The effects of influential parameters including, plate aspect ratio, degree of pit and different ratio of pit depth at each sides of the plate are investigated. The results show that position of pitted surface in respect to applied pressure is the most influential parameter on reduction of dynamic load carrying capacity of pitted plates. By increasing degree of pitting, reduction of dynamic load carrying capacity decrease more.

  19. Heat receiving plates in thermonuclear device

    International Nuclear Information System (INIS)

    Kitamura, Kazunori.

    1988-01-01

    Purpose: To obtain a heat receiving plate structure capable of withstanding sputtering wear and retaining the thermal deformation and residual stress low upon junction and available at a reduced cost. Constitution: Junction structures between heat sinks and armours are the same as usual, whereas high melting armour (for example, made of tungsten) are used at the portion on a heat receiving plate where the thermal load and particle load are higher while materials having a heat expansion coefficient similar to that of the heat sink (stainless steel) are used at the portion where the thermal load and particle load are lower on a heat receiving plate depending on the thermal load and particle load distribution. This can reduce the thermal deformation for the entire divertor heat receiving plate to obtain a heat receiving plate of a good surface dimensional accuracy. (Takahashi, M.)

  20. High thermal load receiving heat plate

    International Nuclear Information System (INIS)

    Shibutani, Jun-ichi; Shibayama, Kazuhito; Yamamoto, Keiichi; Uchida, Takaho.

    1993-01-01

    The present invention concerns a high thermal load heat receiving plate such as a divertor plate of a thermonuclear device. The high thermal load heat receiving plate of the present invention has a cooling performance capable of suppressing the temperature of an armour tile to less than a threshold value of the material against high thermal loads applied from plasmas. Spiral polygonal pipes are inserted in cooling pipes at a portion receiving high thermal loads in the high temperature load heat receiving plate of the present invention. Both ends of the polygonal pipes are sealed by lids. An area of the flow channel in the cooling pipes is thus reduced. Heat conductivity on the cooling surface of the cooling pipes is increased in the high thermal load heat receiving plate having such a structure. Accordingly, temperature elevation of the armour tile can be suppressed. (I.S.)

  1. 2017 Bipolar Plate Workshop Summary Report

    Energy Technology Data Exchange (ETDEWEB)

    Kopasz, John P. [Argonne National Lab. (ANL), Argonne, IL (United States); Benjamin, Thomas G. [Argonne National Lab. (ANL), Argonne, IL (United States); Schenck, Deanna [Argonne National Lab. (ANL), Argonne, IL (United States)

    2017-08-17

    The Bipolar Plate (BP) Workshop was held at USCAR1 in Southfield, Michigan on February 14, 2017 and included 63 participants from industry, government agencies, universities, and national laboratories with expertise in the relevant fields. The objective of the workshop was to identify research and development (R&D) needs, in particular early-stage R&D, for bipolar plates for polymer electrolyte membrane (PEM) fuel cells for transportation applications. The focus of the workshop was on materials, manufacturing, and design aspects of bipolar plates with the goal of meeting DOE’s 2020 bipolar plate targets. Of special interest was the cost target of ≤$3/kW for the bipolar plate.

  2. Pyramidal approach to license plate segmentation

    Science.gov (United States)

    Postolache, Alexandru; Trecat, Jacques C.

    1996-07-01

    Car identification is a goal in traffic control, transport planning, travel time measurement, managing parking lot traffic and so on. Most car identification algorithms contain a standalone plate segmentation process followed by a plate contents reading. A pyramidal algorithm for license plate segmentation, looking for textured regions, has been developed on a PC based system running Unix. It can be used directly in applications not requiring real time. When input images are relatively small, real-time performance is in fact accomplished by the algorithm. When using large images, porting the algorithm to special digital signal processors can easily lead to preserving real-time performance. Experimental results, for stationary and moving cars in outdoor scenes, showed high accuracy and high scores in detecting the plate. The algorithm also deals with cases where many character strings are present in the image, and not only the one corresponding to the plate. This is done by the means of a constrained texture regions classification.

  3. New technique for number-plate recognition

    Science.gov (United States)

    Guo, Jie; Shi, Peng-Fei

    2001-09-01

    This paper presents an alternative algorithm for number plate recognition. The algorithm consists of three modules. Respectively, they are number plate location module, character segmentation module and character recognition module. Number plate location module extracts the number plate from the detected car image by analyzing the color and the texture properties. Different from most license plate location methods, the algorithm has fewer limits to the car size, the car position in the image and the image background. Character segmentation module applies connected region algorithm both to eliminate noise points and to segment characters. Touching characters and broken characters can be processed correctly. Character recognition module recognizes characters with HHIC (Hierarchical Hybrid Integrated Classifier). The system has been tested with 100 images obtained from crossroad and parking lot, etc, where the cars have different size, position, background and illumination. Successful recognition rate is about 92%. The average processing time is 1.2 second.

  4. Neutron radiography using neutron imaging plate

    International Nuclear Information System (INIS)

    Chankow, Nares; Wonglee, Sarinrat

    2008-01-01

    Full text: The aims of this research are to study properties of neutron imaging plate, to obtain a suitable condition for neutron radiography and to use the neutron imaging plate for testing of materials nondestructively. The experiments were carried out by using a neutron beam from the Thai Research Reactor TRR-1/M1 at a power of 1.2 MW. A BAS-ND 2040 FUJI neutron imaging plate and a MX125 Kodak X-ray film/Gadolinium neutron converter screen combination were tested for comparison. It was found that the photostimulated light (PSL) read out of the imaging plate was directly proportional to the exposure time. It was also found that radiography with neutron using the imaging plate was approximately 40 times faster than the conventional neutron radiography using x-ray film/Gd converter screen combination. The sensitivity of the imaging plate to gamma-rays was investigated by using gamma-rays from an 192 Ir and a 60 Co radiographic sources. The imaging plate was found to be 5-6 times less sensitive to gamma-rays than a FUJI BAS-MS 2040 gamma-ray imaging plate. Finally, some specimens were selected to be radiographed with neutrons using the imaging plate and the x-ray film/Gd converter screen combination in comparison to x-rays. Parts containing light elements could be clearly observed by the two neutron radiographic techniques. It could be concluded that the image quality from the neutron imaging plate was comparable to the conventional x-ray film/Gd converter screen combination but the exposure time could be approximately reduced by a factor of 40

  5. Barcoded microchips for biomolecular assays.

    Science.gov (United States)

    Zhang, Yi; Sun, Jiashu; Zou, Yu; Chen, Wenwen; Zhang, Wei; Xi, Jianzhong Jeff; Jiang, Xingyu

    2015-01-20

    Multiplexed assay of analytes is of great importance for clinical diagnostics and other analytical applications. Barcode-based bioassays with the ability to encode and decode may realize this goal in a straightforward and consistent manner. We present here a microfluidic barcoded chip containing several sets of microchannels with different widths, imitating the commonly used barcode. A single barcoded microchip can carry out tens of individual protein/nucleic acid assays (encode) and immediately yield all assay results by a portable barcode reader or a smartphone (decode). The applicability of a barcoded microchip is demonstrated by human immunodeficiency virus (HIV) immunoassays for simultaneous detection of three targets (anti-gp41 antibody, anti-gp120 antibody, and anti-gp36 antibody) from six human serum samples. We can also determine seven pathogen-specific oligonucleotides by a single chip containing both positive and negative controls.

  6. The Iberian Plate: myth or reality?

    Energy Technology Data Exchange (ETDEWEB)

    Canerot, J.

    2016-10-01

    The plate tectonics theory generally leads us to consider that Iberia was an independent plate separated from Europe by the North Pyrenean Fault (NPF). The NPF has been commonly interpreted as a transform fault associated with a huge counterclockwise transverse and rotational movement that allowed the opening of the Bay of Biscay and the relative eastward motion of Iberia during the Mesozoic. According to some interpretations, this movement may have generated an interplate gap several hundreds of km wide, which led to the creation of an oceanic crust during the Late Jurassic and Early Cretaceous. However, field studies recently carried out in the Pyrenees do not support these interpretations. The North Pyrenean Fault (NPF) of Tertiary age is observed in the central and eastern Pyrenees, where pioneering researchers defined it as separating the North Pyrenean Zone from the Axial Zone.However, this fault cannot be identified in the western part of the range to the west of the Ossau valley. Consequently, the geodynamic evolution of Iberia has always been dependent on Europe, especially during the failed oceanic rifting in the Mid-Cretaceous. Indeed, during this period, a central zone of crustal thinning occupied by turbiditic basins separated the European from the Iberian continental crust, with a very localized mantle exhumation found only in the Mauleon basin. Therefore, far from being an interplate range, the Pyrenees can neither be considered as an intraplate unit. We can define this orogenic belt as resulting from the Tertiary tectonic inversion of a Mid-Cretaceous rift system. According to this new interpretation, Iberia would not have been an isolated plate but represented an unstable, outlying part of Europe. Rather than displaying the features of a rigid lithospheric unit with well-defined boundaries, Iberia grouped together different crustal blocks undergoing specific movements at particular times. During the Mesozoic, normal, reverse or strike

  7. Conversion electron Moessbauer spectroscopic studies on the chemical states of surface layers of corroded tin plates and tin-coated iron plates

    International Nuclear Information System (INIS)

    Kato, Akinori; Endo, Kazutoyo; Sano, Hirotoshi

    1980-01-01

    By means of the conversion electron Moessbauer spectroscopy (CEMS), we studied surface layers of ''tin'' plates and tin-coated iron plates corroded by various acids. Transmission Moessbauer spectra and X-ray diffraction patterns were also measured. Metastannic acid was formed, when the ''tin'' plate was corroded by nitric acid solution. In corrosion by phosphoric acid solution, the X-ray diffractometry revealed the formation of tin(IV) pyrophosphate. In corrosion by various organic acid solutions, the formation of oxides was identified by the 119 Sn CEMS, but not by the X-ray diffractometry because of the too thin corrosion layer. In corrosion of tin-coated iron plates, maleic acid, malonic acid, formic acid, and oxalic acid were used. It was determined by CEMS that the corrosion products caused by these acids were tin(IV) oxides, although they could not be identified by the X-ray diffractometry. CEMS also confirmed that the surface of uncorroded tin-coated iron plate was already oxidized by air. Colorimetric determinations of Sn and Fe dissolved from tin-coated iron plates to various acid solutions confirmed that maleic acid had the strongest corrosion effect among the organic acids studied. (author)

  8. A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids

    Energy Technology Data Exchange (ETDEWEB)

    Zipper, Lauren E. [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); Binghamton University, 4400 Vestal Parkway East, Vestal, NY 13902 (United States); Aristide, Xavier [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); North Babylon High School, 1 Phelps Lane North, Babylon, NY 11703 (United States); Bishop, Dylan P. [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); Northport High School, 154 Laurel Hill Road, Northport, NY 11768 (United States); Joshi, Ishita [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); St Augustine Catholic High School, 2188 Rodick Road, Markham, ON L6C 1S3 (Canada); Kharzeev, Julia [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); Earl L. Vandermeulen High School, 350 Old Post Road, Port Jefferson, NY 11777 (United States); Patel, Krishna B. [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); John P. Stevens High School, 855 Grove Avenue, Edison, NJ 08820 (United States); Santiago, Brianna M. [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); Connetquot High School, 190 7th Street, Bohemia, NY 11716 (United States); Joshi, Karan [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); Department of Electronics and Electrical Communication Engineering, PEC University of Technology, Chandigarh (India); Dorsinvil, Kahille [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); Sweet, Robert M. [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); Soares, Alexei S., E-mail: soares@bnl.gov [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States)

    2014-11-28

    This article describes the use of evaporation control lids that are fitted to crystallization plates to improve the reproducibility of trials using as little as 5 nl. The plate lids contain apertures which are large enough for the transfer of protein containing droplets, but small enough to greatly reduce the rate of evaporation during the time needed to prepare the plate. A method is described for using plate lids to reduce evaporation in low-volume vapor-diffusion crystallization experiments. The plate lids contain apertures through which the protein and precipitants were added to different crystallization microplates (the reservoir was filled before fitting the lids). Plate lids were designed for each of these commonly used crystallization microplates. This system minimizes the dehydration of crystallization droplets containing just a few nanolitres of protein and precipitant, and results in more reproducible diffraction from the crystals. For each lid design, changes in the weight of the plates were used to deduce the rate of evaporation under different conditions of temperature, air movement, droplet size and precipitant. For comparison, the state of dehydration was also visually assessed throughout the experiment. Finally, X-ray diffraction methods were used to compare the diffraction of protein crystals that were conventionally prepared against those that were prepared on plates with plate lids. The measurements revealed that the plate lids reduced the rate of evaporation by 63–82%. Crystals grown in 5 nl drops that were set up with plate lids diffracted to higher resolution than similar crystals from drops that were set up without plate lids. The results demonstrate that plate lids can be instrumental for improving few-nanolitre crystallizations.

  9. Voltage-current characteristics of a pin-plate system with different plate configurations

    International Nuclear Information System (INIS)

    Feng, Zhuangbo; Long, Zhengwei

    2013-01-01

    In this paper, the voltage-current (V-I) characteristics of a pin-plate system with four types of collection plate configurations are studied experimentally. The collection plates consider a single metal plate, a metal plate with a fly ash cake layer, a metal plate with a clean filter media and a metal plate with a dirty filter media. The results show that the clean filter media has no obvious effect on the V-I characteristics. But the dirty filter media reduces the current density because of its high resistance. The thick fly ash cake layer increase current density because of the anti-corona effect but the increment is not very obvious.

  10. Finite element analysis of a solar collector plate using two plate geometries

    Directory of Open Access Journals (Sweden)

    Diego Manuel Medina Carril

    2016-09-01

    Full Text Available The thermal behavior of an absorber plate in a solar collector is investigated using finite element analysis. The thermal behavior and efficiency of two absorber plate geometries are studied, using a typical solar collector with a rectangular profile as reference, and a proposed absorber plate with curved geometry. An analysis of the most important parameters involved in the design of the absorber plate was carried out, indicating that the curved geometry of the absorber plate yields an average efficiency ~25% higher than the conventional rectangular geometry. The results suggest that a curved profile made of materials such as aluminum with thermal conductivity higher than 200W/m°C, plate thickness of the order of 2-3mm and with a large density of tubes per unit area of the collector´s plate greatly benefits the thermal efficiency of the solar collector.

  11. Solid-phase peptide quantitation assay using labeled monoclonal antibody and glutaraldehyde fixation

    International Nuclear Information System (INIS)

    Kasprzyk, P.G.; Cuttitta, F.; Avis, I.; Nakanishi, Y.; Treston, A.; Wong, H.; Walsh, J.H.; Mulshine, J.L.

    1988-01-01

    A solid-phase radioimmunoassay utilizing iodinated peptide-specific monoclonal antibody as a detection system instead of labeled peptide has been developed. Regional specific monoclonal antibodies to either gastrin-releasing peptide or gastrin were used as models to validate the general application of our modified assay. Conditions for radioactive labeling of the monoclonal antibody were determined to minimize oxidant damage, which compromises the sensitivity of other reported peptide quantitation assays. Pretreatment of 96-well polyvinyl chloride test plates with a 5% glutaraldehyde solution resulted in consistent retention of sufficient target peptide on the solid-phase matrix to allow precise quantitation. This quantitative method is completed within 1 h of peptide solid phasing. Pretreatment of assay plates with glutaraldehyde increased binding of target peptide and maximized antibody binding by optimizing antigen presentation. The hypothesis that glutaraldehyde affects both peptide binding to the plate and orientation of the peptide was confirmed by analysis of several peptide analogs. These studies indicate that peptide binding was mediated through a free amino group leaving the carboxy-terminal portion of the target peptide accessible for antibody binding. It was observed that the length of the peptide also affects the amount of monoclonal antibody that will bind. Under the optimal conditions, results from quantitation of gastrin-releasing peptide in relevant samples agree well with those from previously reported techniques. Thus, we report here a modified microplate assay which may be generally applied for the rapid and sensitive quantitation of peptide hormones

  12. NDE methods for determining the materials properties of silicon carbide plates

    Science.gov (United States)

    Kenderian, Shant; Kim, Yong; Johnson, Eric; Palusinski, Iwona A.

    2009-08-01

    Two types of SiC plates, differing in their manufacturing processes, were interrogated using a variety of NDE techniques. The task of evaluating the materials properties of these plates was a challenge due to their non-uniform thickness. Ultrasound was used to estimate the Young's Modulus and calculate the thickness profile and Poisson's Ratio of the plates. The Young's Modulus profile plots were consistent with the thickness profile plots, indicating that the technique was highly influenced by the non-uniform thickness of the plates. The Poisson's Ratio is calculated from the longitudinal and shear wave velocities. Because the thickness is cancelled out, the result is dependent only on the time of flight of the two wave modes, which can be measured accurately. X-Ray was used to determine if any density variations were present in the plates. None were detected suggesting that the varying time of flight of the acoustic wave is attributed only to variations in the elastic constants and thickness profiles of the plates. Eddy Current was used to plot the conductivity profile. Surprisingly, the conductivity profile of one type of plates varied over a wide range rarely seen in other materials. The other type revealed a uniform conductivity profile.

  13. Extraction of gadolinium from high flux isotope reactor control plates

    International Nuclear Information System (INIS)

    Kohring, M.W.

    1987-04-01

    Gadolinium-153 is an important radioisotope used in the diagnosis of various bone disorders. Recent medical and technical developments in the detection and cure of osteoporosis, a bone disease affecting an estimated 50 million people, have greatly increased the demand for this isotope. The Oak Ridge National Laboratory (ORNL) has produced 153 Gd since 1980 primarily through the irradiation of a natural europium-oxide powder followed by the chemical separation of the gadolinium fraction from the europium material. Due to the higher demand for 153 Gd, an alternative production method to supplement this process has been investigated. This process involves the extraction of gadolinium from the europium-bearing region of highly radioactive, spent control plates used at the High Flux Isotope Reactor (HFIR) with a subsequent re-irradiation of the extracted material for the production of the 153 Gd. Based on the results of experimental and calculational analyses, up to 25 grams of valuable gadolinium (≥60% enriched in 152 Gd) resides in the europium-bearing region of the HFIR control components of which 70% is recoverable. At a specific activity yield of 40 curies of 153 Gd for each gram of gadolinium re-irradiated, 700 one-curie sources can be produced from each control plate assayed

  14. Vortex Dynamics of Asymmetric Heave Plates

    Science.gov (United States)

    Rusch, Curtis; Maurer, Benjamin; Polagye, Brian

    2017-11-01

    Heave plates can be used to provide reaction forces for wave energy converters, which harness the power in ocean surface waves to produce electricity. Heave plate inertia includes both the static mass of the heave plate, as well as the ``added mass'' of surrounding water accelerated with the object. Heave plate geometries may be symmetric or asymmetric, with interest in asymmetric designs driven by the resulting hydrodynamic asymmetry. Limited flow visualization has been previously conducted on symmetric heave plates, but flow visualization of asymmetric designs is needed to understand the origin of observed hydrodynamic asymmetries and their dependence on the Keulegan-Carpenter number. For example, it is hypothesized that the time-varying added mass of asymmetric heave plates is caused by vortex shedding, which is related to oscillation amplitude. Here, using direct flow visualization, we explore the relationship between vortex dynamics and time-varying added mass and drag. These results suggest potential pathways for more advanced heave plate designs that can exploit vortex formation and shedding to achieve more favorable hydrodynamic properties for wave energy converters.

  15. 3D CFD fluid flow and thermal analyses of a new design of plate heat exchanger

    Directory of Open Access Journals (Sweden)

    Pianko-Oprych Paulina

    2017-03-01

    Full Text Available The paper presents a Computational Fluid Dynamics (CFD numerical study for a new design of a plate heat exchanger with two different flow patterns. The impact of geometric characteristics of the two studied geometries of exchanger plates on the intensification process of heat transfer was considered. The velocity, temperature and pressure distributions along the heat exchanger were examined. The CFD results were validated against experimental data and a good agreement was achieved. The results revealed that geometrical arrangement of the plates strongly influence the fluid flow. An increase in the Reynolds number led to lowering the friction factor value and increasing the pressure drop. The configuration II of the plate heat exchanger resulted in lower outlet hot fluid temperature in comparison with the configuration I, which means improvement of heat transfer.

  16. Image quality and radiation exposure in digital storage plate mammography with magnification technique

    International Nuclear Information System (INIS)

    Fiedler, E.; Aichinger, U.; Boehner, C.; Schulz-Wendtland, R.; Bautz, W.; Saebel, M.

    1999-01-01

    Purpose: Comparison of image quality between digital phosphor storage plate mammography in magnification technique and a conventional film screen system regarding the special aspect of radiation exposure. Materials and Methods: Radiograms of a RMI-mammography phantom were acquired using a conventional film screen system and two digital storage plate systems. Additionally, the radiograms of one digital system were postprocessed emphasizing contrast and included in the comparison. Results: The detectability of details in storage plate mammographies with magnification technique is almost equal to that of film screen mammographies. Thereby, lower radiation exposures were necessary using the digital systems. Conclusions: Based on these results, storage plate mammography in magnification technique is used in clinical routine at our institution. The correct parameters in image postprocessing are of elementary importance for detail detectability. Future studies must show, whether the lower radiation exposure in digital radiograms of the breast, revealing much higher background noise, will allow the same detail detectability as film screen mammographies. (orig.) [de

  17. The influence of retention on the plate height in ion-exchange chromatography

    DEFF Research Database (Denmark)

    Hansen, Ernst; Mollerup, Jørgen

    2004-01-01

    The plate heights for the amino acid tyrosine (anion exchange) and the polypeptide aprotinin (cation exchange) were determined on a porous media (Resource 15) and a get filled media (HyperD 20) at salt concentrations ranging from weak to strong retention. At a constant velocity, measurements showed....... In this article, the rate of mass transfer in the particles is described by three different rate mechanisms, pore diffusion, solid diffusion, and parallel diffusion. The van Deemter equation was used to model the data to determine the mass-transfer properties. The development of the plate height with increasing...... retention revealed a characteristic behavior for each rate mechanism. In the pore diffusion model, the plate height increased toward a constant value at strong retention, while the plate height in the solid diffusion model decreased, approaching a constant value at strong retention. In the parallel...

  18. Optimization of geometrical characteristics of perforated plates

    International Nuclear Information System (INIS)

    Radisavljevic, Igor; Balos, Sebastian; Nikacevic, Milutin; Sidjanin, Leposava

    2013-01-01

    Highlights: ► Perforated plate are tested against 12.7 mm API projectile. ► Perforations similar to the projectile diameter offer more efficient core fracture. ► Larger perforations gave a more efficient core fragmentation. ► SEM microscopy analysis has shown a ductile fracture mode at impact point. - Abstract: In this paper, an attempt was made to design effective non-homogenous armor in form of perforated plate mounted at close distance from basic armor plate. Perforated plate with three perforation diameters: 9, 10 and 11 mm, two ligaments length: 3.5 and 4.5 mm ligaments, set at 0° and 28° angles, were combined to 13 mm basic plate and tested against 12.7 mm API ammunition. It has been shown that larger perforations gave a more efficient core fragmentation, while angled specimens were the only ones that offer full protection against five API shots when the perforated plate was placed at 100 mm from the basic plate. Perforations that are similar in size to the penetrating core diameter offer a more efficient core fracture, leading to a faster fragment separation. This may enable a smaller distance between the add-on perforated and basic plate to be used. Scanning electron microscopy analysis has shown a ductile fracture mode at impact point, with hardness values on plate basic level. On the other hand, a brittle fracture mode with a rise in local hardness measured near impact point is a result of intensive high speed plastic deformation produced by bending stresses. A drop in local hardness measured near impact point, may be the result of intensive cracking that occur due to repeated projectile impact

  19. Immobilization of mesoporous silica particles on stainless steel plates

    International Nuclear Information System (INIS)

    Pasqua, Luigi; Morra, Marco

    2017-01-01

    A preliminary study aimed to the nano-engineering of stainless steel surface is presented. Aminopropyl-functionalized mesoporous silica is covalently and electrostatically anchored on the surface of stainless steel plates. The anchoring is carried out through the use of a nanometric spacer, and two different spacers are proposed (both below 2 nm in size). The first sample is obtained by anchoring to the stainless steel amino functionalized, a glutaryl dichloride spacer. This specie forms an amide linkage with the amino group while the unreacted acyl groups undergo hydrolysis giving a free carboxylic group. The so-obtained functionalized stainless steel plate is used as substrate for anchoring derivatized mesoporous silica particles. The second sample is prepared using 2-bromo-methyl propionic acid as spacer (BMPA). Successively, the carboxylic group of propionic acid is condensed to the aminopropyl derivatization on the external surface of the mesoporous silica particle through covalent bond. In both cases, a continuous deposition (coating thickness is around 10 μm) is obtained, in fact, XPS data do not reveal the metal elements constituting the plate. The nano-engineering of metal surfaces can represent an intriguing opportunity for producing long-term drug release or biomimetic surface.

  20. Circulation shedding in viscous starting flow past a flat plate

    International Nuclear Information System (INIS)

    Nitsche, Monika; Xu, Ling

    2014-01-01

    Numerical simulations of viscous flow past a flat plate moving in the direction normal to itself reveal details of the vortical structure of the flow. At early times, most of the vorticity is attached to the plate. This paper introduces a definition of the shed circulation at all times and shows that it indeed represents vorticity that separates and remains separated from the plate. During a large initial time period, the shed circulation satisfies the scaling laws predicted for self-similar inviscid separation. Various contributions to the circulation shedding rate are presented. The results show that during this initial time period, viscous diffusion of vorticity out of the vortex is significant but appears to be independent of the value of the Reynolds number. At later times, the departure of the shed circulation from its large Reynolds number behaviour is significantly affected by diffusive loss of vorticity through the symmetry axis. A timescale is proposed that describes when the viscous loss through the axis becomes relevant. The simulations provide benchmark results to evaluate simpler separation models such as point vortex and vortex sheet models. A comparison with vortex sheet results is included. (paper)

  1. Distributed sensing signal analysis of deformable plate/membrane mirrors

    Science.gov (United States)

    Lu, Yifan; Yue, Honghao; Deng, Zongquan; Tzou, Hornsen

    2017-11-01

    Deformable optical mirrors usually play key roles in aerospace and optical structural systems applied to space telescopes, radars, solar collectors, communication antennas, etc. Limited by the payload capacity of current launch vehicles, the deformable mirrors should be lightweight and are generally made of ultra-thin plates or even membranes. These plate/membrane mirrors are susceptible to external excitations and this may lead to surface inaccuracy and jeopardize relevant working performance. In order to investigate the modal vibration characteristics of the mirror, a piezoelectric layer is fully laminated on its non-reflective side to serve as sensors. The piezoelectric layer is segmented into infinitesimal elements so that microscopic distributed sensing signals can be explored. In this paper, the deformable mirror is modeled as a pre-tensioned plate and membrane respectively and sensing signal distributions of the two models are compared. Different pre-tensioning forces are also applied to reveal the tension effects on the mode shape and sensing signals of the mirror. Analytical results in this study could be used as guideline of optimal sensor/actuator placement for deformable space mirrors.

  2. Immobilization of mesoporous silica particles on stainless steel plates

    Energy Technology Data Exchange (ETDEWEB)

    Pasqua, Luigi, E-mail: luigi.pasqua@unical.it [University of Calabria, Department of Environmental and Chemical Engineering (Italy); Morra, Marco, E-mail: mmorra@nobilbio.com [Via Valcastellana 26 (Italy)

    2017-03-15

    A preliminary study aimed to the nano-engineering of stainless steel surface is presented. Aminopropyl-functionalized mesoporous silica is covalently and electrostatically anchored on the surface of stainless steel plates. The anchoring is carried out through the use of a nanometric spacer, and two different spacers are proposed (both below 2 nm in size). The first sample is obtained by anchoring to the stainless steel amino functionalized, a glutaryl dichloride spacer. This specie forms an amide linkage with the amino group while the unreacted acyl groups undergo hydrolysis giving a free carboxylic group. The so-obtained functionalized stainless steel plate is used as substrate for anchoring derivatized mesoporous silica particles. The second sample is prepared using 2-bromo-methyl propionic acid as spacer (BMPA). Successively, the carboxylic group of propionic acid is condensed to the aminopropyl derivatization on the external surface of the mesoporous silica particle through covalent bond. In both cases, a continuous deposition (coating thickness is around 10 μm) is obtained, in fact, XPS data do not reveal the metal elements constituting the plate. The nano-engineering of metal surfaces can represent an intriguing opportunity for producing long-term drug release or biomimetic surface.

  3. Chromosome aberration assays in Allium

    Energy Technology Data Exchange (ETDEWEB)

    Grant, W.F.

    1982-01-01

    The common onion (Allium cepa) is an excellent plant for the assay of chromosome aberrations after chemical treatment. Other species of Allium (A. cepa var. proliferum, A. carinatum, A. fistulosum and A. sativum) have also been used but to a much lesser extent. Protocols have been given for using root tips from either bulbs or seeds of Allium cepa to study the cytological end-points, such as chromosome breaks and exchanges, which follow the testing of chemicals in somatic cells. It is considered that both mitotic and meiotic end-points should be used to a greater extent in assaying the cytogenetic effects of a chemical. From a literature survey, 148 chemicals are tabulated that have been assayed in 164 Allium tests for their clastogenic effect. Of the 164 assays which have been carried out, 75 are reported as giving a positive reaction, 49 positive and with a dose response, 1 positive and temperature-related, 9 borderline positive, and 30 negative; 76% of the chemicals gave a definite positive response. It is proposed that the Allium test be included among those tests routinely used for assessing chromosomal damage induced by chemicals.

  4. Transfer coefficients for plate fin and elliptical tube heat exchangers

    International Nuclear Information System (INIS)

    Saboya, S.M.; Saboya, F.E.M.

    1981-01-01

    In order to determine transfer coefficients for plate fin and elliptical tube exchangers, mass transfer experiments have been performed using the naphthalene sublimation technique. By means of the heat-mass transfer analogy, the results can be converted to heat transfer results. The transfer coefficients were compared with those for circular tube exchangers and the comparison revealed no major differences. This is a positive outcome, since the use of elliptical tubes may reduce substantially the pressure drop, without affecting the transfer characteristics.(Author) [pt

  5. Buckling Analysis of Edge Cracked Sandwich Plate

    Directory of Open Access Journals (Sweden)

    Rasha Mohammed Hussein

    2016-07-01

    Full Text Available This work presents mainly the buckling load of sandwich plates with or without crack for different cases. The buckling loads are analyzed experimentally and numerically by using ANSYS 15. The experimental investigation was to fabricate the cracked sandwich plate from stainless steel and PVC to find mechanical properties of stainless steel and PVC such as young modulus. The buckling load for different aspect ratio, crack length, cracked location and plate without crack found. The experimental results were compared with that found from ANSYS program. Present of crack is decreased the buckling load and that depends on crack size, crack location and aspect ratio.

  6. Applications of ion plating in metals fabrication

    International Nuclear Information System (INIS)

    Bell, R.T.; Thompson, J.C.

    1974-01-01

    Use of ion plating at the Oak Ridge Y-12 Plant to solve problems encountered in metals fabrication and processing are discussed. Three typical areas are covered. The first is the use of strike coats on various substrates for subsequent electrodeposition. The second area in which ion plating is shown to contribute to a process is in cold welding or room temperature bonding of metals. The third application involves plating U to promote safe handling, fission-product retention, and corrosion protection in nuclear reactors

  7. Effusion plate using additive manufacturing methods

    Science.gov (United States)

    Johnson, Thomas Edward; Keener, Christopher Paul; Ostebee, Heath Michael; Wegerif, Daniel Gerritt

    2016-04-12

    Additive manufacturing techniques may be utilized to construct effusion plates. Such additive manufacturing techniques may include defining a configuration for an effusion plate having one or more internal cooling channels. The manufacturing techniques may further include depositing a powder into a chamber, applying an energy source to the deposited powder, and consolidating the powder into a cross-sectional shape corresponding to the defined configuration. Such methods may be implemented to construct an effusion plate having one or more channels with a curved cross-sectional geometry.

  8. Fabrication of resistive plate chamber using bakelite

    International Nuclear Information System (INIS)

    Neog, Himangshu; Bhuyan, M.R.; Biswas, S.; Mohanty, B.; Mohanty, Rudranarayan; Rudra, Sharmili; Sahu, P.K.; Sahu, S.

    2014-01-01

    Now a days Resistive Plate Chamber (RPC) is one of the most important detectors in the High Energy Physics (HEP) experiments. RPC is a gas filled detector utilizing a constant and uniform electric field produced between two parallel electrode plates made of a material with high bulk resistivity e.g. glass or bakelite. RPC has good time resolution (1-2 ns) and spatial resolution (∼ cm). The high resistance of RPC plate limits the spark size produced after the ionization of gas due to the passing charged particle. This contribution discusses building of a RPC using bakelite (local sources) and the measurement of the surface resistivity of the detector

  9. Nondestructive diagnosis of multilayer electronic plates

    International Nuclear Information System (INIS)

    Matvienko, A.N.; Savin, D.O.; Yas'ko, A.V.

    1992-01-01

    Methods of non-destructive tomographic investigation into multilayer printed plates using x radiation are described. Mathematic problem setting is given, experimental facility and methods for source data ecquisition are described. A special attention is paid to the consideration of the main factors differing the actual problem setting from the idealized one. Methods for accounting and correction of these factors are described. The efficiency of the approach proposed is demonstrated using the actual problems of reducing separate layers of multilayer printed plate metallization. The method developed is useful when exersizing control over multilayer printed plate production

  10. Reducing risk where tectonic plates collide

    Science.gov (United States)

    Gomberg, Joan S.; Ludwig, Kristin A.

    2017-06-19

    Most of the world’s earthquakes, tsunamis, landslides, and volcanic eruptions are caused by the continuous motions of the many tectonic plates that make up the Earth’s outer shell. The most powerful of these natural hazards occur in subduction zones, where two plates collide and one is thrust beneath another. The U.S. Geological Survey’s (USGS) “Reducing Risk Where Tectonic Plates Collide—A USGS Plan to Advance Subduction Zone Science” is a blueprint for building the crucial scientific foundation needed to inform the policies and practices that can make our Nation more resilient to subduction zone-related hazards.

  11. Joining by plating: optimization of occluded angle

    International Nuclear Information System (INIS)

    Dini, J.W.; Johnson, H.R.; Kan, Y.R.

    1978-11-01

    An empirical method has been developed for predicting the minimum angle required for maximum joint strength for materials joined by plating. This is done through a proposed power law failure function, whose coefficients are taken from ring shear and conical head tensile data for plating/substrate combinations and whose exponent is determined from one set of plated-joint data. Experimental results are presented for Al-Ni-Al (7075-T6) and AM363-Ni-AM363 joints, and the failure function is used to predict joint strengths for Al-Ni-Al (2024-T6), UTi-Ni-UTi, and Be-Ti-Be

  12. Diffusion zinc plating of structural steels

    International Nuclear Information System (INIS)

    Kazakovskaya, Tatiana; Goncharov, Ivan; Tukmakov, Victor; Shapovalov, Vyacheslav

    2004-01-01

    The report deals with the research on diffusion zinc plating of structural steels when replacing their cyanide cadmium plating. The results of the experiments in the open air, in vacuum, in the inert atmosphere, under various temperatures (300 - 500 deg.C) for different steel brands are presented. It is shown that diffusion zinc plating in argon or nitrogen atmosphere ensures obtaining the qualitative anticorrosion coating with insignificant change of mechanical properties of steels. The process is simple, reliable, ecology pure and cost-effective. (authors)

  13. Reconstruction of Northeast Asian Deformation Integrated with Western Pacific Plate Subduction since 200 Ma

    Science.gov (United States)

    Liu, S.; Gurnis, M.; Ma, P.; Zhang, B.

    2017-12-01

    the history of plate motion and subduction and tracing the geological and deformation records in continents play a significant role in revealing the effects of complex plate motions and the interactions of plate boundary forces on plate-mantle coupling and plate motion-intracontinental deformation coupling.

  14. History of internal fixation with plates (part 2): new developments after World War II; compressing plates and locked plates.

    Science.gov (United States)

    Hernigou, Philippe; Pariat, Jacques

    2017-07-01

    The first techniques of operative fracture with plates were developed in the 19th century. In fact, at the beginning these methods consisted of an open reduction of the fracture usually followed by a very unstable fixation. As a consequence, the fracture had to be opened with a real risk of (sometimes lethal) infection, and due to unstable fixation, protection with a cast was often necessary. During the period between World Wars I and II, plates for fracture fixation developed with great variety. It became increasingly recognised that, because a fracture of a long bone normally heals with minimal resorption at the bone ends, this may result in slight shortening and collapse, so a very rigid plate might prevent such collapse. However, as a consequence, delayed healing was observed unless the patient was lucky enough to have the plate break. One way of dealing with this was to use a slotted plate in which the screws could move axially, but the really important advance was recognition of the role of compression. After the first description of compression by Danis with a "coapteur", Bagby and Müller with the AO improved the technique of compression. The classic dynamic compression plates from the 1970s were the key to a very rigid fixation, leading to primary bone healing. Nevertheless, the use of strong plates resulted in delayed union and the osteoporosis, cancellous bone, comminution, and/or pathological bone resulted in some failures due to insufficient stability. Finally, new devices represented by locking plates increased the stability, contributing to the principles of a more biological osteosynthesis while giving enough stability to allow immediate full weight bearing in some patients.

  15. Crack Detection in Armor Plates Using Ultrasonic Techniques

    National Research Council Canada - National Science Library

    Meitzler, Thomas J; Smith, Gregory; Charbeneau, Michelle; Sohn, Euijung; Bienkowski, mary; Wong, Ivan; Meitzler, Allen H

    2008-01-01

    ...) plates for the purpose of crack detection is presented. The amplitudes of the vibrational modes undamaged plates are compared to the vibrational mode amplitudes of damaged plates and are shown to be clearly different...

  16. Comparing Diagnostic Accuracy of Kato-Katz, Koga Agar Plate, Ether-Concentration, and FLOTAC for Schistosoma mansoni and Soil-Transmitted Helminths

    Science.gov (United States)

    Glinz, Dominik; Silué, Kigbafori D.; Knopp, Stefanie; Lohourignon, Laurent K.; Yao, Kouassi P.; Steinmann, Peter; Rinaldi, Laura; Cringoli, Giuseppe; N'Goran, Eliézer K.; Utzinger, Jürg

    2010-01-01

    Background Infections with schistosomes and soil-transmitted helminths exert a considerable yet underappreciated economic and public health burden on afflicted populations. Accurate diagnosis is crucial for patient management, drug efficacy evaluations, and monitoring of large-scale community-based control programs. Methods/Principal Findings The diagnostic accuracy of four copromicroscopic techniques (i.e., Kato-Katz, Koga agar plate, ether-concentration, and FLOTAC) for the detection of Schistosoma mansoni and soil-transmitted helminth eggs was compared using stool samples from 112 school children in Côte d'Ivoire. Combined results of all four methods served as a diagnostic ‘gold’ standard and revealed prevalences of S. mansoni, hookworm, Trichuris trichiura, Strongyloides stercoralis and Ascaris lumbricoides of 83.0%, 55.4%, 40.2%, 33.9% and 28.6%, respectively. A single FLOTAC from stool samples preserved in sodium acetate-acetic acid-formalin for 30 or 83 days showed a higher sensitivity for S. mansoni diagnosis (91.4%) than the ether-concentration method on stool samples preserved for 40 days (85.0%) or triplicate Kato-Katz using fresh stool samples (77.4%). Moreover, a single FLOTAC detected hookworm, A. lumbricoides and T. trichiura infections with a higher sensitivity than any of the other methods used, but resulted in lower egg counts. The Koga agar plate method was the most accurate diagnostic assay for S. stercoralis. Conclusion/Significance We have shown that the FLOTAC method holds promise for the diagnosis of S. mansoni. Moreover, our study confirms that FLOTAC is a sensitive technique for detection of common soil-transmitted helminths. For the diagnosis of S. stercoralis, the Koga agar plate method remains the method of choice. PMID:20651931

  17. Automation of the dicentric chromosome assay and related assays

    International Nuclear Information System (INIS)

    Balajee, Adayabalam S.; Dainiak, Nicholas

    2016-01-01

    Dicentric Chromosome Assay (DCA) is considered to be the 'gold standard' for personalized dose assessment in humans after accidental or incidental radiation exposure. Although this technique is superior to other cytogenetic assays in terms of specificity and sensitivity, its potential application to radiation mass casualty scenarios is highly restricted because DCA is time consuming and labor intensive when performed manually. Therefore, it is imperative to develop high throughput automation techniques to make DCA suitable for radiological triage scenarios. At the Cytogenetic Biodosimetry Laboratory in Oak Ridge, efforts are underway to develop high throughput automation of DCA. Current status on development of various automated cytogenetic techniques in meeting the biodosimetry needs of radiological/nuclear incident(s) will be discussed

  18. Assay strategies and methods for phospholipases

    International Nuclear Information System (INIS)

    Reynolds, L.J.; Washburn, W.N.; Deems, R.A.; Dennis, E.A.

    1991-01-01

    Of the general considerations discussed, the two issues which are most important in choosing an assay are (1) what sensitivity is required to assay a particular enzyme and (2) whether the assay must be continuous. One can narrow the options further by considering substrate availability, enzyme specificity, assay convenience, or the presence of incompatible side reactions. In addition, the specific preference of a particular phospholipase for polar head group, micellar versus vesicular substrates, and anionic versus nonionic detergents may further restrict the options. Of the many assays described in this chapter, several have limited applicability or serious drawbacks and are not commonly employed. The most commonly used phospholipase assays are the radioactive TLC assay and the pH-stat assay. The TLC assay is probably the most accurate, sensitive assay available. These aspects often outweigh the disadvantages of being discontinuous, tedious, and expensive. The radioactive E. coli assay has become popular recently as an alternative to the TLC assay for the purification of the mammalian nonpancreatic phospholipases. The assay is less time consuming and less expensive than the TLC assay, but it is not appropriate when careful kinetics are required. Where less sensitivity is needed, or when a continuous assay is necessary, the pH-stat assay is often employed. With purified enzymes, when free thiol groups are not present, a spectrophotometric thiol assay can be used. This assay is ∼ as sensitive as the pH-stat assay but is more convenient and more reproducible, although the substrate is not available commercially. Despite the many assay choices available, the search continues for a convenient, generally applicable assay that is both sensitive and continuous

  19. Structure phenomena in the bond zone of explosively bonded plates

    International Nuclear Information System (INIS)

    Livne, Z.

    1979-12-01

    In the bond areas of couples of explosively bonded plates, there are often zones, generally designated as ''molten pockets'', which have undergone melting and solidification. The object of the present study was to investigate molten pockets, which have a decisive effect on bond quality. The experimental samples for the study were chosen in consideration of the mutual behaviour of the plates constituting the couples, according to their equilibrium phase diagrams. To facilitate the investigation, large plates were bonded under conditions that enabled to to obtain wavy bond zones that included relatively large molten pockets. To clarify the complex nature of molten pockets and their surroundings, a wide variety of methods were employed. It was found that the shape and composition of molten pockets largely depend upon the mechanism of formation of both the bond wave and the molten pockets. It was also found that the composition of molten pockets is not homogeneous, which is manifest in the modification of the composition of the pockets, the solidification morphology, the phases, which have been identified by X-ray diffraction, and the bond strenght and hardness. Moreover, the different solidification morphologies revealed by metallography were found to depend upon the types of plates bonded, the bonding conditions and the location of pockets in the wavy interface. For molten pockets, cooling rates of 10 4 to 10 5 (degC/sec) have been deduced from interdendritic spacing, and found to be in good agreement with calculations after a mathematical model. It seems that the fast cooling rates and the steep temperature gradients are at the origin of the particular solidification phenomena observed in molten pockets

  20. The comet assay: assessment of in vitro and in vivo DNA damage.

    Science.gov (United States)

    Bajpayee, Mahima; Kumar, Ashutosh; Dhawan, Alok

    2013-01-01

    Rapid industrialization and pursuance of a better life have led to an increase in the amount of chemicals in the environment, which are deleterious to human health. Pesticides, automobile exhausts, and new chemical entities all add to air pollution and have an adverse effect on all living organisms including humans. Sensitive test systems are thus required for accurate hazard identification and risk assessment. The Comet assay has been used widely as a simple, rapid, and sensitive tool for assessment of DNA damage in single cells from both in vitro and in vivo sources as well as in humans. Already, the in vivo comet assay has gained importance as the preferred test for assessing DNA damage in animals for some international regulatory guidelines. The advantages of the in vivo comet assay are its ability to detect DNA damage in any tissue, despite having non-proliferating cells, and its sensitivity to detect genotoxicity. The recommendations from the international workshops held for the comet assay have resulted in establishment of guidelines. The in vitro comet assay conducted in cultured cells and cell lines can be used for screening large number of compounds and at very low concentrations. The in vitro assay has also been automated to provide a high-throughput screening method for new chemical entities, as well as environmental samples. This chapter details the in vitro comet assay using the 96-well plate and in vivo comet assay in multiple organs of the mouse.

  1. Degradation behaviour of LAE442-based plate-screw-systems in an in vitro bone model.

    Science.gov (United States)

    Wolters, Leonie; Besdo, Silke; Angrisani, Nina; Wriggers, Peter; Hering, Britta; Seitz, Jan-Marten; Reifenrath, Janin

    2015-04-01

    The use of absorbable implant materials for fixation after bone fracture helps to avoid a second surgery for implant removal and the risks and costs involved. Magnesium (Mg) is well known as a potential metallic material for degradable implants. The aim of the present in vitro study was to evaluate if degradable LAE442-based magnesium plate-screw-systems are suitable candidates for osteosynthesis implants in load-bearing bones. The corrosion behaviour was tested concerning the influence of different surface treatments, coatings and screw torques. Steel plates and screws of the same size served as control. Plates without special treatment screwed on up to a specified torque of 15cNm or 7cNm, NaOH treated plates (15cNm), magnesium fluoride coated plates (15cNm) and steel plates as control (15cNm) were examined in pH-buffered, temperature-controlled SBF solution for two weeks. The experimental results indicate that the LAE442 plates and screws coated with magnesium fluoride revealed a lower hydrogen evolution in SBF solution as well as a lower weight loss and volume decrease in μ-computed tomography (μCT). The nanoindentation and SEM/EDX measurements at several plate areas showed no significant differences. Summarized, the different screw torques did not affect the corrosion behaviour differently. Also the NaOH treatment seemed to have no essential influence on the degradation kinetics. The plates coated with magnesium fluoride showed a decreased corrosion rate. Hence, it is recommended to consider this coating for the next in vivo study. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. The Interaction between the Plane Wave and the Plate with Limited Height in Soil

    Directory of Open Access Journals (Sweden)

    N.A. Lokteva

    2017-03-01

    Full Text Available A solution of the two-dimensional task on interaction between the harmonic wave and the plate with the limited height in soil has been provided. The plate surrounded on both sides with the half-spaces filled with soil medium has been used as a vibro-absorbing obstacle. The mechanical behavior of the plate has been described by S.P. Timoshenko's shift model and the mechanical behavior of soil by a linear elasticity theory equation. The main purpose of the paper is to determine the total acceleration vector field inducted by the penetrated and radiated waves in the second half-space. The mathematical formulation of the task includes a model of upcoming wave, soil medium and plate movement equation, infinity conditions, and conditions of soil contact with obstacle. Conditions of free slip have been taken as the contact conditions between the soil and the obstacle. We have considered a closed system of equations, which includes wave equations for scalar and vector potentials, elasticity theory equations for soil mediums, Koshi's relations, physical law, and plate movement equation. The boundary conditions for the plate correspond to a hinged support. To solve this task, all functions have been expanded in trigonometric series that allowed to obtain potential values in the coefficients of the series. To define the integrations constants, the contact conditions between the obstacle and soil have been used. On the basis of the revealed potentials, we have defined displacements on the boundary between the plate and soil and in other points of the second half-space. The vibro-absorbing properties of the plate have been investigated depending on the frequency of the harmonic wave falling on the plate. From the practical point of view, this task is related to protection of buildings from vibrations formed at a distance from underground railways.

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

    Science.gov (United States)

    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.

  4. Validation of a KHV antibody enzyme-linked immunosorbent assay (ELISA).

    Science.gov (United States)

    Bergmann, S M; Wang, Q; Zeng, W; Li, Y; Wang, Y; Matras, M; Reichert, M; Fichtner, D; Lenk, M; Morin, T; Olesen, N J; Skall, H F; Lee, P-Y; Zheng, S; Monaghan, S; Reiche, S; Fuchs, W; Kotler, M; Way, K; Bräuer, G; Böttcher, K; Kappe, A; Kielpinska, J

    2017-11-01

    Koi herpesvirus (KHV) causes KHV disease (KHVD). The virus is highly contagious in carp or koi and can induce a high mortality. Latency and, in some cases, a lack of signs presents a challenge for virus detection. Appropriate immunological detection methods for anti-KHV antibodies have not yet been fully validated for KHV. Therefore, it was developed and validated an enzyme-linked immunosorbent assay (ELISA) to detect KHV antibodies. The assay was optimized with respect to plates, buffers, antigens and assay conditions. It demonstrated high diagnostic and analytical sensitivity and specificity and was particularly useful at the pond or farm levels. Considering the scale of the carp and koi industry worldwide, this assay represents an important practical tool for the indirect detection of KHV, also in the absence of clinical signs. © 2017 John Wiley & Sons Ltd.

  5. Flaw behavior in mechanically loaded clad plates

    International Nuclear Information System (INIS)

    Iskander, S.K.; Robinson, G.C.; Oland, C.B.

    1989-01-01

    A small crack near the inner surface of clad nuclear reactor pressure vessels is an important consideration in the safety assessment of the structural integrity of the vessel. Four-point bend tests on large plate specimens, conforming to ASTM specification for pressure vessel plates, alloy steels, quenched and tempered, Mn-Mo and Mn-Mo-Ni (A533) grade B six clad and two unclad with stainless steels 308, 309 and 312 weld wires, were performed to determine the effect of cladding upon the propagation of small surface cracks subjected to stress states. Results indicated that the tough surface layer composed of cladding and/or heat-affected zone has enhanced the load-bearing capacity of plates under conditions where unclad plates have ruptured. The results are interpreted in terms of fracture mechanics. The behavior of flaws in clad reactor pressure vessels is examined in the light of the test results. 11 refs., 8 figs., 2 tabs

  6. Introduction to Plate Boundaries and Natural Hazards

    NARCIS (Netherlands)

    Duarte, João C.; Schellart, Wouter P.

    2016-01-01

    A great variety of natural hazards occur on Earth, including earthquakes, volcanic eruptions, tsunamis, landslides, floods, fires, tornadoes, hurricanes, and avalanches. The most destructive of these hazards, earthquakes, tsunamis, and volcanic eruptions, are mostly associated with tectonic plate

  7. Vibro-acoustic analysis of composite plates

    International Nuclear Information System (INIS)

    Sarigül, A S; Karagözlü, E

    2014-01-01

    Vibro-acoustic analysis plays a vital role on the design of aircrafts, spacecrafts, land vehicles and ships produced from thin plates backed by closed cavities, with regard to human health and living comfort. For this type of structures, it is required a coupled solution that takes into account structural-acoustic interaction which is crucial for sensitive solutions. In this study, coupled vibro-acoustic analyses of plates produced from composite materials have been performed by using finite element analysis software. The study has been carried out for E-glass/Epoxy, Kevlar/Epoxy and Carbon/Epoxy plates with different ply angles and numbers of ply. The effects of composite material, ply orientation and number of layer on coupled vibro-acoustic characteristics of plates have been analysed for various combinations. The analysis results have been statistically examined and assessed

  8. Vibro-acoustic analysis of composite plates

    Science.gov (United States)

    Sarigül, A. S.; Karagözlü, E.

    2014-03-01

    Vibro-acoustic analysis plays a vital role on the design of aircrafts, spacecrafts, land vehicles and ships produced from thin plates backed by closed cavities, with regard to human health and living comfort. For this type of structures, it is required a coupled solution that takes into account structural-acoustic interaction which is crucial for sensitive solutions. In this study, coupled vibro-acoustic analyses of plates produced from composite materials have been performed by using finite element analysis software. The study has been carried out for E-glass/Epoxy, Kevlar/Epoxy and Carbon/Epoxy plates with different ply angles and numbers of ply. The effects of composite material, ply orientation and number of layer on coupled vibro-acoustic characteristics of plates have been analysed for various combinations. The analysis results have been statistically examined and assessed.

  9. Modeling the hydrodynamics of Phloem sieve plates

    DEFF Research Database (Denmark)

    Jensen, Kaare Hartvig; Mullendore, Daniel Leroy; Holbrook, Noel Michele

    2012-01-01

    Sieve plates have an enormous impact on the efficiency of the phloem vascular system of plants, responsible for the distribution of photosynthetic products. These thin plates, which separate neighboring phloem cells, are perforated by a large number of tiny sieve pores and are believed to play...... understood. We propose a theoretical model for quantifying the effect of sieve plates on the phloem in the plant, thus unifying and improving previous work in the field. Numerical simulations of the flow in real and idealized phloem channels verify our model, and anatomical data from 19 plant species...... a crucial role in protecting the phloem sap from intruding animals by blocking flow when the phloem cell is damaged. The resistance to the flow of viscous sap in the phloem vascular system is strongly affected by the presence of the sieve plates, but the hydrodynamics of the flow through them remains poorly...

  10. New high voltage parallel plate analyzer

    International Nuclear Information System (INIS)

    Hamada, Y.; Kawasumi, Y.; Masai, K.; Iguchi, H.; Fujisawa, A.; Abe, Y.

    1992-01-01

    A new modification on the parallel plate analyzer for 500 keV heavy ions to eliminate the effect of the intense UV and visible radiations, is successfully conducted. Its principle and results are discussed. (author)

  11. Plate performance in liquid-liquid extraction

    International Nuclear Information System (INIS)

    Wadkins, R.P.

    1984-01-01

    The relative effectiveness of perforated, nozzle, and burred plates from a capacity and extraction standpoint were studied in a pulsed liquid-liquid extraction system. The experiments were conducted in a 3.8 x 10 -2 m diameter column using a mixture of aluminum nitrate, nitric acid, and uranyl nitrate as the aqueous phase, and tributyl phosphate dissolved in AMSCO 125-90 W as the organic phase. The uranium was extracted from the aqueous phase to the organic phase. A standard cartridge was made for each type of plate and consisted of an assembly of plates spaced 5.08 x 10 -2 m apart. Each plate had 3.2 x 10 -3 m holes spaced on 6.1 x 10 -3 -m centers, and contained 23% free area. 16 references, 4 figures, 1 table

  12. Heavy gauge plates for nuclear application

    International Nuclear Information System (INIS)

    Cheviet, A.; Roux, J.-H.

    1977-01-01

    The production of energy from nuclear sources leads to the building of very large vessels working under pressure at elevated temperatures, requiring very thick steel plate (from 50 mm to 300 mm). The plates necessary for the production of these vessels have to be as large as possible in order to reduce the length of welds on the vessels. Those two requirements lead to the manufacture of heavy products from 10 to 80 tons unit weight. These products are special, because their fabrication requires very big facilities and also extremely high quality of the steel. The main points are: high cleanliness; properties as homogeneous as possible. The tests carried out on industrially produced plates (especially on a plate of 200 mm thick show the level of quality that can be reached [fr

  13. Microchannel plate assembly parameters with micron gaps

    International Nuclear Information System (INIS)

    Demchenkova, A.A.

    1987-01-01

    Performance of chevron microchannel plate assembly with 5 and 15 μm gaps between them has been investigated. The assembly is placed into a vacuum chamber under pressure -6 Torr and irradiated by neutral He and Ar atom beams with 1.5 and 3 keV energies as well as by ultraviolet photons with 147 nm wave length. Dependence of the gain and amplitude resolution on power voltage in plates are measured. The results obtained have shown that microchannel plates permit to obtain the gain up to 3x10 7 and amplitude resolution up to 30% when detecting both atomic particles and ultraviolet photons. The assembly can be effectively used in those cases when it is necessary to use microchannel plates with curved channels

  14. White Light Photorefractive Phase Zone Plates

    International Nuclear Information System (INIS)

    Yuan-Mei, Gao; Si-Min, Liu

    2008-01-01

    Incoherent white light from an incandescent source is employed to fabricate volume phase zone plates in LiNbO 3 : Fe, for the first time to our knowledge, which can guide and modulate the input white light or laser light. The diffractive efficiency of the white light volume phase zone plates fabricated can reach as high as 12%. In addition, we test the volume phase zone plates by a probe beam and find that the volume phase zone plate is present in the direction perpendicular to the c-axis and absent in the direction parallel to the c-axis. This directly proves the existence of photovoltaic photorefractive anisotropy of white light

  15. Buckling Response of Thick Functionally Graded Plates

    Directory of Open Access Journals (Sweden)

    BOUAZZA MOKHTAR

    2014-11-01

    Full Text Available In this paper, the buckling of a functionally graded plate is studied by using first order shear deformation theory (FSDT. The material properties of the plate are assumed to be graded continuously in the direction of thickness. The variation of the material properties follows a simple power-law distribution in terms of the volume fractions of constituents. The von Karman strains are used to construct the equilibrium equations of the plates subjected to two types of thermal loading, linear temperature rise and gradient through the thickness are considered. The governing equations are reduced to linear differential equation with boundary conditions yielding a simple solution procedure. In addition, the effects of temperature field, volume fraction distributions, and system geometric parameters are investigated. The results are compared with the results of the no shear deformation theory (classic plate theory, CPT.

  16. COMPARATIVE STUDY ON MILK CASEIN ASSAY METHODS

    Directory of Open Access Journals (Sweden)

    RODICA CĂPRIłĂ

    2008-05-01

    Full Text Available Casein, the main milk protein was determined by different assay methods: the gravimetric method, the method based on the neutralization of the NaOH excess used for the casein precipitate solving and the method based on the titration of the acetic acid used for the casein precipitation. The last method is the simplest one, with the fewer steps, and also with the lowest error degree. The results of the experiment revealed that the percentage of casein from the whole milk protein represents between 72.6–81.3% in experiment 1, between 73.6–81.3% in experiment 2 and between 74.3–81% in experiment 3.

  17. 21 CFR 866.4800 - Radial immunodiffusion plate.

    Science.gov (United States)

    2010-04-01

    ...) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents § 866.4800 Radial immunodiffusion plate. (a) Identification. A radial immunodiffusion plate for clinical use...

  18. Infection control in digital intraoral radiography: evaluation of microbiological contamination of photostimulable phosphor plates in barrier envelopes.

    Science.gov (United States)

    MacDonald, David S; Waterfield, J Douglas

    2011-01-01

    The detectors (both solid-state sensors and photostimulable phosphor [PSP] plates) used for digital intraoral radiography cannot be autoclaved, and barriers are typically used to prevent the spread of infection. The aim of this study was to determine the effectiveness of a barrier envelope system for PSP plates. Disinfected PSP plates were aseptically inserted into barrier envelopes and placed in a periapical location. One PSP plate was placed in each of 28 patients, and 12 plates in each of 2 volunteers (D.S.M., J.D.W.). After retrieval, each PSP plate was removed from its barrier envelope, immersed in trypticase soy broth and aliquots were plated on trypticase soy agar. Bacterial colonies were counted 2 days later. Fifty-two PSP plates in barrier envelopes were evaluated for contamination. Quality assurance of the PSP plates before clinical placement revealed defects in the integrity of 4 barrier envelopes, caused by forceps-related damage or failure to achieve a uniform seal. These defects allowed substantial contamination. Contamination also occurred as a result of failure to extract the PSP plate from the barrier envelope cleanly. Of the 44 barriers with no obvious defects that were placed by either final-year dental students or a radiologist, only 3 allowed bacterial contamination of the PSP plate. Detectors contained in barrier envelopes remain a potential source of contamination. PSP plates must be disinfected between removal from a contaminated barrier envelope and placement in a new barrier envelope. In addition, placement into the barrier envelope should ideally be carried out under aseptic conditions. Finally, the integrity of each sealed barrier envelope must be verified visually before release to the clinic.

  19. Inert gas handling in ion plating systems

    International Nuclear Information System (INIS)

    Goode, A.R.; Burden, M.St.J.

    1979-01-01

    The results of an investigation into the best methods for production and monitoring of the inert gas environment for ion plating systems are reported. Work carried out on Pirani gauges and high pressure ion gauges for the measurement of pressures in the ion plating region (1 - 50mtorr) and the use of furnaces for cleaning argon is outlined. A schematic of a gas handling system is shown and discussed. (UK)

  20. Selenium plating of aluminium and nickel surfaces

    International Nuclear Information System (INIS)

    Qureshi, N.; Shams, N.; Kamal, A.; Ashraf, A.

    1993-01-01

    Selenium exhibits photovoltaic and photoconductive properties. This makes selenium useful in the production of photocells, exposure meters for photographic use, in solar cells, etc. In commerce, selenium coated surfaces are extensively used as photo receptive drums in the xerography machines for reproducing documents. Laboratory experiments were designed to obtain selenium plating on different materials. Of the various electrodes tested for cathodic deposition, anodized aluminum and nickel plated copper were found to give good results. (author)

  1. Investigation of vertebral ''end plate sclerosis''

    International Nuclear Information System (INIS)

    Lee, S.W.; Mathie, A.G.; Jackson, J.E.; Hughes, S.P.F.

    2001-01-01

    To evaluate the association between vertebral ''end plate sclerosis'' and neck pain. A retrospective study was carried out of lateral cervical spine radiographs with a Picture Archive and Communication System (PACS). Two hundred patients' files were randomly assessed, comprising four equal groups, A to D. The mean ages of the patients were 62±7.4 years, 61±7.5 years, 40±5.6 years and 23±5.6 years respectively. In group A, all patients had symptoms of neck pain and a radiographic diagnosis of ''end plate sclerosis'' of the cervical spine. In groups B to D, asymptomatic patients were recruited and their age groups were 50-69, 30-49 and 10-29 years respectively. Using the PACS, the radiographic density and the sagittal diameter, thickness and area of the end plates at the C5 level were measured. Results and conclusions: No significant differences were found in the radiographic density of the end plates either between the symptomatic and asymptomatic groups (groups A and B), or between different age groups (groups B, C and D). A significant increase in end plate area and thickness was found, however, in both group B (P<0.005) and group C (P<0.01) in comparison with group D. This indicates that the extent of end plate sclerosis increases with age. Our results suggest that the radiographic density of cervical vertebral end plates correlates neither with neck pain nor with increasing age. The radiological sign of ''end plate sclerosis'' may be over-reported, further limiting its value in the assessment of patients with cervical spondylosis. (orig.)

  2. Detection of extracellular proteases from microorganisms on agar plates

    Directory of Open Access Journals (Sweden)

    Alane Beatriz Vermelho

    1996-12-01

    Full Text Available We present herein an improved assay for detecting the presence of extracellular proteases from microorganisms on agar plates. Using different substrates (gelatin, BSA, hemoglobin incorporated into the agar and varying the culture medium composition, we were able to detect proteolytic activities from Pseudomonas aeruginosa, Micrococcus luteus and Serratia marcescens as well as the influence that these components displayed in the expression of these enzymes. For all microorganisms tested we found that in agar-BHI or yeast extract medium containing gelatin the sensitivity of proteinase detection was considerably greater than in BSA-agar or hemoglobin-agar. However, when BSA or hemoglobin were added to the culture medium, there was an increase in growth along with a marked reduction in the amount of proteinase production. In the case of M. luteus the incorporation of glycerol in BHI or yeast extract gelatin-agar induced protease liberation. Our results indicate that the technique described here is of value for detecting extracellular proteases directly in the culture medium, by means of a qualitative assay, simple, inexpensive, straight forward method to assess the presence of the proteolytic activity of a given microorganism colony with great freedom in substrate selection.

  3. Evaluation of e-liquid toxicity using an open-source high-throughput screening assay

    Science.gov (United States)

    Keating, James E.; Zorn, Bryan T.; Kochar, Tavleen K.; Wolfgang, Matthew C.; Glish, Gary L.; Tarran, Robert

    2018-01-01

    The e-liquids used in electronic cigarettes (E-cigs) consist of propylene glycol (PG), vegetable glycerin (VG), nicotine, and chemical additives for flavoring. There are currently over 7,700 e-liquid flavors available, and while some have been tested for toxicity in the laboratory, most have not. Here, we developed a 3-phase, 384-well, plate-based, high-throughput screening (HTS) assay to rapidly triage and validate the toxicity of multiple e-liquids. Our data demonstrated that the PG/VG vehicle adversely affected cell viability and that a large number of e-liquids were more toxic than PG/VG. We also performed gas chromatography–mass spectrometry (GC-MS) analysis on all tested e-liquids. Subsequent nonmetric multidimensional scaling (NMDS) analysis revealed that e-liquids are an extremely heterogeneous group. Furthermore, these data indicated that (i) the more chemicals contained in an e-liquid, the more toxic it was likely to be and (ii) the presence of vanillin was associated with higher toxicity values. Further analysis of common constituents by electron ionization revealed that the concentration of cinnamaldehyde and vanillin, but not triacetin, correlated with toxicity. We have also developed a publicly available searchable website (www.eliquidinfo.org). Given the large numbers of available e-liquids, this website will serve as a resource to facilitate dissemination of this information. Our data suggest that an HTS approach to evaluate the toxicity of multiple e-liquids is feasible. Such an approach may serve as a roadmap to enable bodies such as the Food and Drug Administration (FDA) to better regulate e-liquid composition. PMID:29584716

  4. A comparison of assays measuring the viability of Legionella ...

    Science.gov (United States)

    Background: The relatively high prevalence of Legionella pneumophila in premise plumbing systems has been widely reported. Published reports indicate Legionella has a comparatively high resistance to chlorine and moreover has the ability to grow in phagocytic amoeba which could provide additional protection in chlorinated drinking water distribution systems. Copper-Silver (Cu-Ag) ionization treatment systems are commercially available for use in large building water systems to help control the risks from Legionella bacteria. The objectives of this study were to develop and optimize Legionella viability assays and use them to investigate the viability of Legionella bacteria after exposure to water treated with coppper and silver ions. Methods: Log phase L. pneumophila cells were used in all experiments and were generated by incubation at 35C for 48 hours in buffered yeast extract broth. Viability assays used included plating on buffered charcoal yeast extract agar to determine the number of culturable cells and treating cells with propidium monoazide (PMA) or ethidium monoazide (EMA) followed by quantitative PCR targeting mip gene of L. pneumophila. The qPCR viability assays were optimized using L. pneumophila inactivated by heat treatment at 65C for 60 min. The effectiveness of Cu-Ag ionization treatment was studied by inoculating L. pneumonia at 105 CFU/mL in water collected directly from a building water system that employed this technology and incubat

  5. Desomorphine Screening Using Commercial Enzyme-Linked Immunosorbent Assays.

    Science.gov (United States)

    Winborn, Jessica; Kerrigan, Sarah

    2017-06-01

    Desomorphine ("Krokodil") is a semi-synthetic opioid that has drawn attention as a recreational drug, particularly in Russia, neighboring former Soviet Republics, Eastern and Central Europe. It has no accepted medicinal uses and is currently a schedule I drug in the United States. In clandestine environments, desomorphine is synthesized from codeine using red phosphorous, hydroiodic acid and gasoline. Residual starting materials in illicit preparations have been associated with severe dermatological effects and extensive tissue necrosis. Desomorphine is not well studied, and there are limited reports concerning its pharmacology or detection in biological matrices. Immunoassays are widely relied upon for both antemortem and postmortem toxicology screening. Although desomorphine is an opioid of the phenanthrene-type, its ability to bind to conventional opioid antibodies has not been described. In this report we describe the cross-reactivity of desomorphine using six commercially available enzyme-linked immunosorbent assays (Immunalysis Opiates Direct ELISA, Immunalysis Oxycodone/Oxymorphone Direct ELISA, Randox Opiate ELISA, OraSure Technologies OTI Opiate Micro-plate EIA, Neogen Opiate Group ELISA and Neogen Oxycodone/Oxymorphone ELISA). Cross-reactivites were highly variable between assays, ranging from 77 to desomorphine than those directed towards oxycodone. The Immunalysis Opiates Direct ELISA produced the greatest cross-reactivity, although several of the assays evaluated produced cross-reactivity of a sufficient magnitude to be effective for desomorphine screening. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  6. [Experimental study on carbon fiber reinforced plastic plate--analysis of stabilizing force required for plate].

    Science.gov (United States)

    Iizuka, H

    1990-11-01

    Plates currently in use for the management of bone fracture made of metal present with various problems. We manufactured carbon fiber reinforced plastic (CFRP) plates from Pyrofil T/530 puriplegs overlaid at cross angles of +/- 10 degrees, +/- 20 degrees, and +/- 30 degrees for trial and carried out an experimental study on rabbit tibiofibular bones using 316L stainless steel plates of comparable shape and size as controls. The results indicate the influence of CFRP plate upon cortical bone was milder than that of stainless steel plate, with an adequate stabilizing force for the repair of fractured rabbit tibiofibular bones. CFRP has the advantages over metals of being virtually free from corrosion and fatigue, reasonably radiolucent and able to meet a wide range of mechanical requirements. This would make CFRP plate quite promising as a new devices of treating fracture of bones.

  7. Modeling the hydrodynamics of phloem sieve plates

    Directory of Open Access Journals (Sweden)

    Kaare Hartvig Jensen

    2012-07-01

    Full Text Available Sieve plates have an enormous impact on the efficiency of the phloem vascular system of plants, responsible for the distribution of photosynthetic products. These thin plates, which separate neighboring phloem cells, are perforated by a large number of tiny sieve pores and are believed to play a crucial role in protecting the phloem sap from intruding animals by blocking flow when the phloem cell is damaged. The resistance to the flow of viscous sap in the phloem vascular system is strongly affected by the presence of the sieve plates, but the hydrodynamics of the flow through them remains poorly understood. We propose a theoretical model for quantifying the effect of sieve plates on the phloem in the plant, thus unifying and improving previous work in the field. Numerical simulations of the flow in real and idealized phloem channels verify our model, and anatomical data from 19 plant species are investigated. We find that the sieve plate resistance is correlated to the cell lumen resistance, and that the sieve plate and the lumen contribute almost equally to the total hydraulic resistance of the phloem translocation pathway.

  8. Image simulation for automatic license plate recognition

    Science.gov (United States)

    Bala, Raja; Zhao, Yonghui; Burry, Aaron; Kozitsky, Vladimir; Fillion, Claude; Saunders, Craig; Rodríguez-Serrano, José

    2012-01-01

    Automatic license plate recognition (ALPR) is an important capability for traffic surveillance applications, including toll monitoring and detection of different types of traffic violations. ALPR is a multi-stage process comprising plate localization, character segmentation, optical character recognition (OCR), and identification of originating jurisdiction (i.e. state or province). Training of an ALPR system for a new jurisdiction typically involves gathering vast amounts of license plate images and associated ground truth data, followed by iterative tuning and optimization of the ALPR algorithms. The substantial time and effort required to train and optimize the ALPR system can result in excessive operational cost and overhead. In this paper we propose a framework to create an artificial set of license plate images for accelerated training and optimization of ALPR algorithms. The framework comprises two steps: the synthesis of license plate images according to the design and layout for a jurisdiction of interest; and the modeling of imaging transformations and distortions typically encountered in the image capture process. Distortion parameters are estimated by measurements of real plate images. The simulation methodology is successfully demonstrated for training of OCR.

  9. Benchmark for license plate character segmentation

    Science.gov (United States)

    Gonçalves, Gabriel Resende; da Silva, Sirlene Pio Gomes; Menotti, David; Shwartz, William Robson

    2016-09-01

    Automatic license plate recognition (ALPR) has been the focus of many researches in the past years. In general, ALPR is divided into the following problems: detection of on-track vehicles, license plate detection, segmentation of license plate characters, and optical character recognition (OCR). Even though commercial solutions are available for controlled acquisition conditions, e.g., the entrance of a parking lot, ALPR is still an open problem when dealing with data acquired from uncontrolled environments, such as roads and highways when relying only on imaging sensors. Due to the multiple orientations and scales of the license plates captured by the camera, a very challenging task of the ALPR is the license plate character segmentation (LPCS) step, because its effectiveness is required to be (near) optimal to achieve a high recognition rate by the OCR. To tackle the LPCS problem, this work proposes a benchmark composed of a dataset designed to focus specifically on the character segmentation step of the ALPR within an evaluation protocol. Furthermore, we propose the Jaccard-centroid coefficient, an evaluation measure more suitable than the Jaccard coefficient regarding the location of the bounding box within the ground-truth annotation. The dataset is composed of 2000 Brazilian license plates consisting of 14000 alphanumeric symbols and their corresponding bounding box annotations. We also present a straightforward approach to perform LPCS efficiently. Finally, we provide an experimental evaluation for the dataset based on five LPCS approaches and demonstrate the importance of character segmentation for achieving an accurate OCR.

  10. Waveguiding in supported phononic crystal plates

    International Nuclear Information System (INIS)

    Vasseur, J; Hladky-Hennion, A-C; Deymier, P; Djafari-Rouhani, B; Duval, F; Dubus, B; Pennec, Y

    2007-01-01

    We investigate, with the help of the finite element method, the existence of absolute band gaps in the band structure of a free-standing phononic crystal plate and of a phononic crystal slab deposited on a substrate. The two-dimensional phononic crystal is constituted by a square array of holes drilled in an active piezoelectric (PZT5A or AlN) matrix. For both matrix materials, an absolute band gap occurs in the band structure of the free-standing plate provided the thickness of the plate is on the order of magnitude of the lattice parameter. When the plate is deposited on a Si substrate, the absolute band gap still remains when the matrix of the phononic crystal is made of PZT5A. The AlN phononic crystal plate losses its gap when supported by the Si substrate. In the case of the PZT5A matrix, we also study the possibility of localized modes associated with a linear defect created by removing one row of air holes in the deposited phononic crystal plate

  11. Quantitative assessment of growth plate activity

    International Nuclear Information System (INIS)

    Harcke, H.T.; Macy, N.J.; Mandell, G.A.; MacEwen, G.D.

    1984-01-01

    In the immature skeleton the physis or growth plate is the area of bone least able to withstand external forces and is therefore prone to trauma. Such trauma often leads to premature closure of the plate and results in limb shortening and/or angular deformity (varus or valgus). Active localization of bone seeking tracers in the physis makes bone scintigraphy an excellent method for assessing growth plate physiology. To be most effective, however, physeal activity should be quantified so that serial evaluations are accurate and comparable. The authors have developed a quantitative method for assessing physeal activity and have applied it ot the hip and knee. Using computer acquired pinhole images of the abnormal and contralateral normal joints, ten regions of interest are placed at key locations around each joint and comparative ratios are generated to form a growth plate profile. The ratios compare segmental physeal activity to total growth plate activity on both ipsilateral and contralateral sides and to adjacent bone. In 25 patients, ages 2 to 15 years, with angular deformities of the legs secondary to trauma, Blount's disease, and Perthes disease, this technique is able to differentiate abnormal segmental physeal activity. This is important since plate closure does not usually occur uniformly across the physis. The technique may permit the use of scintigraphy in the prediction of early closure through the quantitative analysis of serial studies

  12. Nondestructive assay of sale materials

    International Nuclear Information System (INIS)

    Rodenburg, W.W.; Fleissner, J.G.

    1981-01-01

    This paper covers three primary areas: (1) reasons for performing nondestructive assay on SALE materials; (2) techniques used; and (3) discussion of investigators' revised results. The study shows that nondestructive calorimetric assay of plutonium offers a viable alternative to traditional wet chemical techniques. For these samples, the precision ranged from 0.4 to 0.6% with biases less than 0.2%. Thus, for those materials where sampling errors are the predominant source of uncertainty, this technique can provide improved accuracy and precision while saving time and money as well as reducing the amount of liquid wastes to be handled. In addition, high resolution gamma-ray spectroscopy measurements of solids can provide isotopic analysis data in a cost effective and timely manner. The timeliness of the method can be especially useful to the plant operator for production control and quality control measurements

  13. Comet Assay in Cancer Chemoprevention.

    Science.gov (United States)

    Santoro, Raffaela; Ferraiuolo, Maria; Morgano, Gian Paolo; Muti, Paola; Strano, Sabrina

    2016-01-01

    The comet assay can be useful in monitoring DNA damage in single cells caused by exposure to genotoxic agents, such as those causing air, water, and soil pollution (e.g., pesticides, dioxins, electromagnetic fields) and chemo- and radiotherapy in cancer patients, or in the assessment of genoprotective effects of chemopreventive molecules. Therefore, it has particular importance in the fields of pharmacology and toxicology, and in both environmental and human biomonitoring. It allows the detection of single strand breaks as well as double-strand breaks and can be used in both normal and cancer cells. Here we describe the alkali method for comet assay, which allows to detect both single- and double-strand DNA breaks.

  14. Radioreceptor assay for somatomedin A

    Energy Technology Data Exchange (ETDEWEB)

    Takano, K [Tokyo Women' s Medical Coll. (Japan)

    1975-04-01

    Measurement method of somatomedian A by radioreceptor assay using the human placenta membrane was described and discussed. Binding rate of /sup 125/I-somatomedin A to its receptors was studied under various conditions of time and temperature of the incubation, and pH of the system. The influence of somatomedin A, porcine insulin, and porcine calcitonin, on /sup 125/I-somatomedin A bound receptors was studied, and these hormones showed the competitive binding to somatomedin A receptors in some level. The specificity, recovery rate, and clinical applications of somatomedin A were also discussed. Radioreceptor assay for somatomedine A provided easier, faster, and more accurate measurements than conventional bioassay. This technique would be very useful to study somatomedin A receptor and functions of insulin.

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

    International Nuclear Information System (INIS)

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

    1988-01-01

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

  16. A radiolabel-release microwell assay for proteolytic enzymes present in cell culture media

    International Nuclear Information System (INIS)

    Rucklidge, G.J.; Milne, G.

    1990-01-01

    A modified method for the measurement of proteolytic enzyme activity in cell culture-conditioned media has been developed. Using the release of 3H-labeled peptides from 3H-labeled gelatin the method is performed in microwell plates. The substrate is insolubilized and attached to the wells by glutaraldehyde treatment, thus eliminating the need for a precipitation step at the end of the assay. The assay is sensitive, reproducible, and convenient for small sample volumes. The effect of different protease inhibitors on activity can be assessed rapidly allowing an early characterization of the enzyme. It can also be adapted to microplate spectrophotometric analysis by staining residual substrate with Coomassie blue

  17. A Possible Differentially Shortened Strike-slip Plate Boundary: the Okhotsk Plate Example.

    Science.gov (United States)

    Hindle, D.; Egorov, V.; Mackey, K. G.; Fujita, K.

    2004-12-01

    The Okhotsk plate has been postulated based on a combination of GPS geodetic inversions (REVEL1), seimsicity, geologic and lineament data. Lying between the North American and Eurasian plates, its northwestern corner would appear to be undergoing compression in a scissors motion between the two bounding plates. Extrusion tectonics along multiple, large strike-slip faults within the Okhotsk plate itself have been suggested to allow the escape of material away from the apex of Eurasia-North America. The plate boundary between Okhotsk and North America has been suggested to be diffuse, based on widely scattered minor seismicity. However, the large, left lateral, Ulakhan fault has also been suggested as a candidate plate boundary. We present field geological and geomorphological evidence of the partitioning of deformation between the Ulakhan fault, and several parallel and oblique, linked faults. The Ulakhan fault strand appears to have a maximum displacement of 24 km based on river valley offsets and closing large pull apart basins. Some of the displacement from the Ulakhan fault appears relayed into the plate margin along oblique trending, thrust/oblique slip faults. Estimated shortening over these faults is equivalent to the amount of shortening relayed into the plate margin from the plate boundary. There may be several thrust/oblique slip faults along the Ulakhan fault, which leads to the interesting situation of a segmented, strike-slip plate boundary being actively shortened in a margin parallel direction. This may be the result of postulated extrusion of the Okhotsk plate due to North America/Eurasia convergence. Such a situation would have important consequences for the interpretation of GPS data in a plate tectonic context.

  18. Automation of a Nile red staining assay enables high throughput quantification of microalgal lipid production.

    Science.gov (United States)

    Morschett, Holger; Wiechert, Wolfgang; Oldiges, Marco

    2016-02-09

    Within the context of microalgal lipid production for biofuels and bulk chemical applications, specialized higher throughput devices for small scale parallelized cultivation are expected to boost the time efficiency of phototrophic bioprocess development. However, the increasing number of possible experiments is directly coupled to the demand for lipid quantification protocols that enable reliably measuring large sets of samples within short time and that can deal with the reduced sample volume typically generated at screening scale. To meet these demands, a dye based assay was established using a liquid handling robot to provide reproducible high throughput quantification of lipids with minimized hands-on-time. Lipid production was monitored using the fluorescent dye Nile red with dimethyl sulfoxide as solvent facilitating dye permeation. The staining kinetics of cells at different concentrations and physiological states were investigated to successfully down-scale the assay to 96 well microtiter plates. Gravimetric calibration against a well-established extractive protocol enabled absolute quantification of intracellular lipids improving precision from ±8 to ±2 % on average. Implementation into an automated liquid handling platform allows for measuring up to 48 samples within 6.5 h, reducing hands-on-time to a third compared to manual operation. Moreover, it was shown that automation enhances accuracy and precision compared to manual preparation. It was revealed that established protocols relying on optical density or cell number for biomass adjustion prior to staining may suffer from errors due to significant changes of the cells' optical and physiological properties during cultivation. Alternatively, the biovolume was used as a measure for biomass concentration so that errors from morphological changes can be excluded. The newly established assay proved to be applicable for absolute quantification of algal lipids avoiding limitations of currently established

  19. The OECD validation program of the H295R steroidogenesis assay for the identification of in vitro inhibitors and inducers of testosterone and estradiol production. Phase 2: Inter-laboratory pre-validation studies

    DEFF Research Database (Denmark)

    Hecker, Markus; Hollert, Henner; Cooper, Ralph

    2007-01-01

    plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. Results. All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different...

  20. Resorbable versus titanium plates for orthognathic surgery.

    Science.gov (United States)

    Agnihotry, Anirudha; Fedorowicz, Zbys; Nasser, Mona; Gill, Karanjot S

    2017-10-04

    Recognition of some of the limitations of titanium plates and screws used for the fixation of bones has led to the development of plates manufactured from bioresorbable materials. Whilst resorbable plates appear to offer clinical advantages over metal plates in orthognathic surgery, concerns remain about the stability of fixation and the length of time required for their degradation and the possibility of foreign body reactions. This review compares the use of titanium versus bioresorbable plates in orthognathic surgery and is an update of the Cochrane Review first published in 2007. To compare the effects of bioresorbable fixation systems with titanium systems used during orthognathic surgery. Cochrane Oral Health's Information Specialist searched the following databases: Cochrane Oral Health's Trials Register (to 20 January 2017); the Cochrane Central Register of Controlled Trials (CENTRAL; 2016, Issue 11) in the Cochrane Library (searched 20 January 2017); MEDLINE Ovid (1946 to 20 January 2017); and Embase Ovid (1980 to 20 January 2017). We searched the US National Institutes of Health Ongoing Trials Register ClinicalTrials.gov (clinicaltrials.gov; searched 20 January 2017), and the World Health Organization International Clinical Trials Registry Platform (searched 20 January 2017) for ongoing trials. No restrictions were placed on the language or date of publication when searching the electronic databases. Randomised controlled trials comparing bioresorbable versus titanium fixation systems used for orthognathic surgery in adults. Two review authors independently screened the results of the electronic searches, extracted data and assessed the risk of bias of the included studies. We resolved disagreement by discussion. Clinical heterogeneity between the included trials precluded pooling of data, and only a descriptive summary is presented. This review included two trials, involving 103 participants, one comparing titanium with resorbable plates and screws and

  1. Assay of vitamin B12

    International Nuclear Information System (INIS)

    Tovey, K.C.; Carrick, D.T.

    1982-01-01

    A radioassay is described for vitamin B12 which involves denaturing serum protein binding proteins with alkali. In the denaturation step a dithiopolyol and cyanide are used and in the intrinsic factor assay step a vitamin B12 analogue such as cobinamide is used to bind with any remaining serum proteins. The invention also includes a kit in which the dithiopolyol is provided in admixture with the alkali. The dithiopolyol may be dithiothreitol or dithioerythritol. (author)

  2. Immobilisation of barley aleurone layers enables parallelisation of assays and analysis of transient gene expression in single cells

    DEFF Research Database (Denmark)

    Zor, Kinga; Mark, Christina; Heiskanen, Arto

    2017-01-01

    at a single time point. By immobilising barley aleurone layer tissue on polydimethylsiloxane pillars in the lid of a multiwell plate, continuous monitoring of living tissue is enabled using multiple non-destructive assays in parallel. Cell viability and menadione reducing capacity were monitored in the same...

  3. A high throughput screening assay for identifying glycation inhibitors on MALDI-TOF target.

    Science.gov (United States)

    Zhang, Qiuting; Tu, Zongcai; Wang, Hui; Fan, Liangliang; Huang, Xiaoqin; Xiao, Hui

    2015-03-01

    The Maillard reaction plays an important role in the food industry, however, the deleterious effects generated by the advanced glycation end-products (AGEs) have been well recognized. Many efforts have been made to seek new AGE inhibitors, in particular those natural ones without adverse effect. We have developed a rapid, mass spectrometry based, on-plate screening assay for novel AGE inhibitors. The glycation reaction, inhibition feedback as well as the subsequent MALDI mass spectrometric analysis occurred on one single MALDI plate. At 1:10 M ratio of peptide to sugar, as little as 4h incubation time allowed the screening test to be ready for analysis. DSP, inhibition and IC50 were calculated to evaluate selected inhibitors and resulting inhibition efficiencies were consistent with available references. We demonstrated that this method provide a potential high throughput screening assay to analyze and identify the anti-glycation agents. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Development and Fit-for-Purpose Validation of a Soluble Human Programmed Death-1 Protein Assay.

    Science.gov (United States)

    Ni, Yan G; Yuan, Xiling; Newitt, John A; Peterson, Jon E; Gleason, Carol R; Haulenbeek, Jonathan; Santockyte, Rasa; Lafont, Virginie; Marsilio, Frank; Neely, Robert J; DeSilva, Binodh; Piccoli, Steven P

    2015-07-01

    Programmed death-1 (PD-1) protein is a co-inhibitory receptor which negatively regulates immune cell activation and permits tumors to evade normal immune defense. Anti-PD-1 antibodies have been shown to restore immune cell activation and effector function-an exciting breakthrough in cancer immunotherapy. Recent reports have documented a soluble form of PD-1 (sPD-1) in the circulation of normal and disease state individuals. A clinical assay to quantify sPD-1 would contribute to the understanding of sPD-1-function and facilitate the development of anti-PD-1 drugs. Here, we report the development and validation of a sPD-1 protein assay. The assay validation followed the framework for full validation of a biotherapeutic pharmacokinetic assay. A purified recombinant human PD-1 protein was characterized extensively and was identified as the assay reference material which mimics the endogenous analyte in structure and function. The lower limit of quantitation (LLOQ) was determined to be 100 pg/mL, with a dynamic range spanning three logs to 10,000 pg/mL. The intra- and inter-assay imprecision were ≤15%, and the assay bias (percent deviation) was ≤10%. Potential matrix effects were investigated in sera from both normal healthy volunteers and selected cancer patients. Bulk-prepared frozen standards and pre-coated Streptavidin plates were used in the assay to ensure consistency in assay performance over time. This assay appears to specifically measure total sPD-1 protein since the human anti-PD-1 antibody, nivolumab, and the endogenous ligands of PD-1 protein, PDL-1 and PDL-2, do not interfere with the assay.

  5. Assay of ribulose bisphosphate carboxylase

    International Nuclear Information System (INIS)

    Pike, C.; Berry, J.

    1987-01-01

    Assays of ribulose bisphosphate carboxylase (rubisco) can be used to illustrate many properties of photosynthetic systems. Many different leaves have been assayed with this standard procedure. The tissue is ground with a mortar and pestle in extraction buffer. The supernatant after centrifugation is used as the source of enzyme. Buffer, RuBP, [ 14 C]-NaHCO 3 , and enzyme are combined in a scintillation vial; the reaction is run for 1 min at 30 0 . The acid-stable products are counted. Reproducibility in student experiments has been excellent. The assay data can be combined with analyses of leaf properties such as fresh and dry weight, chlorophyll and protein content, etc. Students have done projects such as the response of enzyme to temperature and to various inhibitors. They also report on the use of a transition state analog, carboxyarabinitol bisphosphate, to titrate the molar concentration of rubisco molecules (active sites) in an enzyme sample. Thus, using crude extracts the catalytic activity of a sample can be compared to the absolute quantity of enzyme or to the turnover number

  6. Theroretical modelling of the plate-tubes coupling in the hydroelasticity of the perforated plates

    International Nuclear Information System (INIS)

    Dzhupanov, V.A.; Manoach, E.S.

    1983-01-01

    In the previous investigations on the perforated plate hydroelasticity the problem of the plates-tubes-liquid interaction in the process of the general structural vibration is stated. But the interaction of the vibrating plates with the tubes, passing through them, is taken into account considering the tubes only as absolutely rigid supports. This is one of the possible technical realizations. In the present article the case when the tubes are taking part in the plate motion (vibration) is studied. Two circular perforated plates are supported by the absolutely rigid wall of the modelled roundcircular reactor barrel. The distance between the plates is given. They are connected by tubes, passing through, and clamped into the perforation holes. The plates and the tubes are made by any elastic HOOKIAN material. The volume between the two plates and outwardly to the tubes, but intrinsically of the barrel is filled by ideal, compressible and heavy liquid. Evidently the liquid volume is multiconnected one. The free vibration of the whole system is considered with the purposes: i) to give a theoretical model of the plates-tubes-liquid interaction including governing equations and boundary conditions; ii) to trace the solution of the eigen-value problem for the modelled structure; iii) to underline the engineering sides of the modelling process. (orig./GL)

  7. A novel microculture kinetic assay (MiCK assay) for malignant cell growth and chemosensitivity.

    Science.gov (United States)

    Kravtsov, V D

    1994-01-01

    The THERMOmax microplate reader was adapted for monitoring the growth kinetics of human leukaemic OCI/AML-2 and mouse tumour J-774.1 cell lines in continuous culture. Fluid evaporation from wells, CO2 escape and contamination were prevented by hermetic sealing of the microcultures in wells of a 96-well microplate, thus enabling the cells to grow exponentially for 72 h under the conditions of the incubated microplate reader. For both OCI/AML-2 cells, which grow in suspension, and adherent J-774.1 cells, a linear correlation was demonstrated between the number of unstained cells seeded in a given microplate well and the optical density (OD) of that well. Therefore, the OD/time curve of the culture could be deemed to be its growth curve. By the use of the linear fit equation, the actual number of the cells in the wells was computable at any time point of the assay. In the chemosensitivity test, an inhibitory effect of ARA-C on the growth of the cells could be estimated by viewing of the growth curves plotted on the screen. The maximum kinetic rates (Vmax) of the curves in the control and the ARA-C-treated wells were compared, yielding a growth inhibition index (GII). Comparison of results of the kinetic chemosensitivity assay with those of a [3H]thymidine incorporation assay revealed that the novel assay is suitable for precise quantitation of the cell chemosensitivity, is more informative and has the added technical advantage of performance without recourse to radioactive or chemically hazardous substances.

  8. External vortex pumping by oscillating plate arrays of mayfly nymphs

    Science.gov (United States)

    Sensenig, Andrew; Kiger, Ken; Shultz, Jeffrey

    2009-11-01

    Mayfly nymphs are aquatic insects, many of which can generate ventilation currents by beating two linear arrays of external plate-like gills. The oscillation Reynolds number associated with the gill motion changes with animal size, varying from Re ˜ 2 to 50 depending on age and species. Thus mayflies provide a novel system model for studying ontogenetic changes in pumping mechanisms associated with transitions from a more viscous- to inertia-dominated flow. Observation of the 3-D kinematics of the gill motion of the species C. triangulifer reveal that the mayfly makes a transition in stroke motion when Re>5, with a corresponding shift in mean flow from the ventral to the dorsal direction. Time-resolved PIV measurements within the inter-gill space reveal the basic elements of the flow consist of vortex rings generated by the strokes of the individual gills. For the larger Re case, the phasing of the plate motion generates a complex array of small vortices that interact to produce an intermittent dorsally directed jet. For Restroke kinematics. Thus we argue the transition in the kinematics is a reflection of a single mechanism adapted over the traversed Re range, rather than a shift to a completely new mechanism. This work is supported by the NSF under grant CBET-0730907.

  9. Determination of Aldehyde Dehydrogenase (ALDH Isozymes in Human Cancer Samples - Comparison of Kinetic and Immunochemical Assays

    Directory of Open Access Journals (Sweden)

    Dorota Borecka

    2002-12-01

    Full Text Available A fluorimetric assay of aldehyde dehydrogenase isozymes, based on naphthaldehyde oxidation, is compared with Western Blotting analysis on several clinical samples obtained from surgery. The comparison reveals qualitatively good correlation of ALDH1A1 isozyme detection with two methods and somewhat worse on ALDH3A1 assay.

  10. Quantitative digital image analysis of chromogenic assays for high throughput screening of alpha-amylase mutant libraries.

    Science.gov (United States)

    Shankar, Manoharan; Priyadharshini, Ramachandran; Gunasekaran, Paramasamy

    2009-08-01

    An image analysis-based method for high throughput screening of an alpha-amylase mutant library using chromogenic assays was developed. Assays were performed in microplates and high resolution images of the assay plates were read using the Virtual Microplate Reader (VMR) script to quantify the concentration of the chromogen. This method is fast and sensitive in quantifying 0.025-0.3 mg starch/ml as well as 0.05-0.75 mg glucose/ml. It was also an effective screening method for improved alpha-amylase activity with a coefficient of variance of 18%.

  11. Measurement of immunoglobulin production by peripheral blood mononuclear cells in vitro using a solid-phase immunoradiometric assay

    International Nuclear Information System (INIS)

    Roffe, L.M.; Maini, R.N.; Cohen, M.L.; Meretey, K.

    1981-01-01

    A simple solid-phase immunoradiometric assay for IgG and IgM is described. Supernatants from lymphocyte cultures are incubated in microtitre plates which have been precoated with anti-IgG or anti-IgM. Subsequent binding of 125 I-labelled anti-immunoglobulin is measured and IgG and IgM in supernatants are estimated from the standard curve constructed for each assay. The assay is specific for human IgG and IgM, is able to detect nanogram amounts and offers advantages over other techniques for evaluating in vitro lymphocyte function. (Auth.)

  12. Crustal deformation and volcanism at active plate boundaries

    Science.gov (United States)

    Geirsson, Halldor

    geometry and secular rates across the plate boundary segments, reveals a deep magma chamber under Hekla and gives a geodetic estimate of the current location of the North-America Eurasian plate boundary in south Iceland. Different geometries were tested for Hekla's magma chamber: spherical, horizontally elongated ellipsoidal, and pipe-like magma chambers. The data could not reliably distinguish the actual geometry; however, all three models indicate magma accumulation near the Moho (˜20-25 km) under Hekla. The February -- March 2000 eruption of Hekla gave another opportunity to image the magmatic system. In Chapter 5, I used co-eruptive GPS and InSAR displacements, borehole strain, and tilt measurements to jointly invert for co-eruptive deformation associated with the 2000 eruption and found a depth of approximately 20 km for the magma chamber, in accordance with my previous results. Telica is a highly seismically active volcano in Nicaragua. The seismicity is mostly of shallow (earthquake per minute averaged over 24 hours, but overall trends in seismic activity, as observed since 1993, do not have an obvious correlation with eruptive activity. This variability causes difficulties for hazard monitoring of Telica. Telica erupted in a small (VEI 2) explosive eruption in 2011. Eruptions of this style and size seem to occur on decadal time scales at Telica. In Chapter 3, I used an extensive multidisciplinary data set consisting of seismic and GPS data, multivariate ash analysis, SO2 measurements, fumarole temperatures, and visual observations, to show that the eruption was essentially an amagmatic eruption of hydrothermally altered materials from the conduit, and that short-term sealing of hydrothermal pathways led to temporary pressure build-up, resulting in the explosions. No significant crustal deformation was detected before or during the eruption, in accordance with low (earthquake offshore El Salvador was the largest event to rupture this segment of the subduction

  13. Technical note: Time-resolved immunofluorometric assay for growth hormone in ruminants

    DEFF Research Database (Denmark)

    Løvendahl, P.; Adamsen, J.; Lund, Regina Teresa

    2003-01-01

    for 4 h at 25degreesC. Plates were then washed six times, incubated for 5 to 10 min with 250 muL of enhancement solution, and fluorescence read with a time-resolved fluorometer. The sensitivity of the assay was 0.1 ng/mL, and the working range was 0.2 to 200 ng/mL. Recovery of quantitative amounts...

  14. Generation of orientation tools for automated zebrafish screening assays using desktop 3D printing

    OpenAIRE

    Wittbrodt, Jonas N.; Liebel, Urban; Gehrig, Jochen

    2014-01-01

    Background The zebrafish has been established as the main vertebrate model system for whole organism screening applications. However, the lack of consistent positioning of zebrafish embryos within wells of microtiter plates remains an obstacle for the comparative analysis of images acquired in automated screening assays. While technical solutions to the orientation problem exist, dissemination is often hindered by the lack of simple and inexpensive ways of distributing and duplicating tools. ...

  15. A membrane-based ELISA assay for the herbicide Isoproturon in soil samples

    OpenAIRE

    Baskeyfield, Damian E. H.; Davis, Frank; Magan, Naresh; Tothill, Ibtisam E.

    2012-01-01

    A membrane based enzyme linked immunosorbent assay (MELISA) for the detection of a common herbicide, isoproturon is described. A heterogeneous competitive ELISA was the format chosen for isoproturon detection. An immunoassay system with a horseradish peroxidase (HRP) labeled polyclonal antibody preparation was developed and characterized before suitable sensitivity and selectivity for isoproturon were attained. After development as a microtiter plate immunoassay, the system was transferred to...

  16. An automated multiplex specific IgE assay system using a photoimmobilized microarray.

    Science.gov (United States)

    Ito, Yoshihiro; Moritsugu, Nozomi; Matsue, Takahisa; Mitsukoshi, Kiyomi; Ayame, Hirohito; Okochi, Norihiko; Hattori, Hideshi; Tashiro, Hideo; Sato, Sakura; Ebisawa, Motohiro

    2012-11-15

    An automated microarray diagnostic system for specific IgE using photoimmobilized allergen has been developed. Photoimmobilization is useful for preparing microarrays, where various types of biological components are covalently immobilized on a plate. Because the immobilization is based on a photo-induced radical cross-linking reaction, it does not require specific functional groups on the immobilized components. Here, an aqueous solution of a photoreactive poly(ethylene glycol)-based polymer was spin-coated on a plate, and an aqueous solution of each allergen was microspotted on the coated plate and allowed to dry in air. Finally, the plate was irradiated with an ultraviolet lamp for covalent immobilization. An automated machine using these plates was developed for the assay of antigen-specific IgE. Initially, the patient serum was added to the microarray plate, and after reaction of the microspotted allergen with IgE, the adsorbed IgE was detected by a peroxidase-conjugated anti-IgE-antibody. The chemical luminescence intensity of the substrate decomposed by the peroxidase was automatically detected using a sensitive charge-coupled device camera. All the allergens were immobilized stably using this method, which was used to screen for allergen-specific IgE. The results were comparable with those using conventional specific IgE. Using this system, six different allergen-specific IgE were assayed using 10 μL of serum within a period of 20 min. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Plate tectonics in the late Paleozoic

    Directory of Open Access Journals (Sweden)

    Mathew Domeier

    2014-05-01

    Full Text Available As the chronicle of plate motions through time, paleogeography is fundamental to our understanding of plate tectonics and its role in shaping the geology of the present-day. To properly appreciate the history of tectonics—and its influence on the deep Earth and climate—it is imperative to seek an accurate and global model of paleogeography. However, owing to the incessant loss of oceanic lithosphere through subduction, the paleogeographic reconstruction of ‘full-plates’ (including oceanic lithosphere becomes increasingly challenging with age. Prior to 150 Ma ∼60% of the lithosphere is missing and reconstructions are developed without explicit regard for oceanic lithosphere or plate tectonic principles; in effect, reflecting the earlier mobilistic paradigm of continental drift. Although these ‘continental’ reconstructions have been immensely useful, the next-generation of mantle models requires global plate kinematic descriptions with full-plate reconstructions. Moreover, in disregarding (or only loosely applying plate tectonic rules, continental reconstructions fail to take advantage of a wealth of additional information in the form of practical constraints. Following a series of new developments, both in geodynamic theory and analytical tools, it is now feasible to construct full-plate models that lend themselves to testing by the wider Earth-science community. Such a model is presented here for the late Paleozoic (410–250 Ma together with a review of the underlying data. Although we expect this model to be particularly useful for numerical mantle modeling, we hope that it will also serve as a general framework for understanding late Paleozoic tectonics, one on which future improvements can be built and further tested.

  18. Postirradiation Examination Of U3O8-AL Plate Type Dispersion Fuel Element

    International Nuclear Information System (INIS)

    Nasution-Hasbullah; Sugondo; Amin, D.L.; Siti-Amini

    1996-01-01

    Postirradiation examination of plate type spent fuel element RIE-01 has been carried out in order to observer its physical changes and performance under irradiation in the reactor. The irradiation has been time more than two years with a declared burnup of 51.04 %. The examination included visual and dimensional measurement, measurement of burn-up distribution, wipe test and metallographic analysis. The results showed that all fuel plates retained their integrity. The colour changes were occurred on most of the plates significant suggesting that it was generated from the oxide layer formation. From gamma-scanning examination it could be deducted that the highest burn-up distribution of the plate was at position of 30 cm from the bottom. A more homogeneous distribution was found in the middle plate of the bundle. The increased plate thickness, as revealed by dimensional measurements as in agreement with the burn-up distribution pattern. Despite the changes observed in could be concluded that all changes occurred were still within the allowable limits and therefore it can recommended that an increase of the burn-up level above 51,04 % is still quite possible

  19. Flow over a traveling wavy foil with a passively flapping flat plate

    Science.gov (United States)

    Liu, Nansheng; Peng, Yan; Liang, Youwen; Lu, Xiyun

    2012-05-01

    Flow over a traveling wavy foil with a passively flapping flat plate has been investigated using a multiblock lattice Boltzmann equation and the immersed boundary method. The foil undergoes prescribed undulations in the lateral direction and the rigid flat plate has passive motion determined by the fluid structure interaction. This simplified model is used to study the effect of the fish caudal fin and its flexibility on the locomotion of swimming animals. The flexibility of the caudal fin is modeled by a torsion spring acting about the pivot at the conjuncture of the wavy foil and the flat plate. The study reveals that the passively oscillating flat plate contributes half of the propulsive force. The flexibility, represented by the nondimensional natural frequency F, plays a very important role in the movement and propulsive force generation of the whole body. When the plate is too flexible, the drag force is observed. As the flat plate becomes more rigid, the propulsive force that is generated when the undulation is confined to last part of the wavy foil becomes larger. The steady movement occurs at F=5. These results are consistent with the observations of some swimming animals in nature.

  20. Effect of Plate Curvature on Blast Response of Structural Steel Plates

    Science.gov (United States)

    Veeredhi, Lakshmi Shireen Banu; Ramana Rao, N. V.; Veeredhi, Vasudeva Rao

    2018-04-01

    In the present work an attempt is made, through simulation studies, to determine the effect of plate curvature on the blast response of a door structure made of ASTM A515 grade 50 steel plates. A door structure with dimensions of 5.142 m × 2.56 m × 10 mm having six different radii of curvatures is analyzed which is subjected to blast load. The radii of curvature investigated are infinity (flat plate), 16.63, 10.81, 8.26, 6.61 and 5.56 m. In the present study, a stand-off distance of 11 m is considered for all the cases. Results showed that the door structure with smallest radius of curvature experienced least plastic deformation and yielding when compared to a door with larger radius of curvature with same projected area. From the present Investigation, it is observed that, as the radius of curvature of the plate increases, the deformation mode gradually shifts from indentation mode to flexural mode. The plates with infinity and 16.63 m radius of curvature have undergone flexural mode of deformation and plates with 6.61 and 5.56 m radius of curvature undergo indentation mode of deformation. Whereas, mixed mode of deformation that consists of both flexural and indentation mode of deformations are seen in the plates with radius of curvature 10.81 and 8.26 m. As the radius of curvature of the plate decreases the ability of the plate to mitigate the effect the blast loads increased. It is observed that the plate with smaller radius of curvature deflects most of the blast energy and results in least indentation mode of deformation. The most significant observation made in the present investigation is that the strain energy absorbed by the steel plate gets reduced to 1/3 rd when the radius of curvature is approximately equal to the stand-off distance which could be the critical radius of curvature.