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Sample records for rapid cze method

  1. Application of CZE Method in Routine Analysis for Determination of B-Complex Vitamins in Pharmaceutical and Veterinary Preparations

    OpenAIRE

    Marina Franco; Renata Jasionowska; Elisa Salvatore

    2012-01-01

    A competitive CZE method for quality control analysis of multivitamin preparations and veterinary products containing B-group vitamins was developed. Vitamins of interest are thiamine hydrochloride (B1), thiamine monophosphate chloride (B1a), riboflavine (B2), riboflavine-5′monophosphate (B2a), nicotinamide (B3), d-pantothenic acid calcium salt (B5), pyridoxine hydrochloride (B6), folic acid (B9), and 4-aminobenzoic acid (B10). These analytes were separated optimizing the experimental conditi...

  2. Application of CZE Method in Routine Analysis for Determination of B-Complex Vitamins in Pharmaceutical and Veterinary Preparations

    Directory of Open Access Journals (Sweden)

    Marina Franco

    2012-01-01

    Full Text Available A competitive CZE method for quality control analysis of multivitamin preparations and veterinary products containing B-group vitamins was developed. Vitamins of interest are thiamine hydrochloride (B1, thiamine monophosphate chloride (B1a, riboflavine (B2, riboflavine-5′monophosphate (B2a, nicotinamide (B3, d-pantothenic acid calcium salt (B5, pyridoxine hydrochloride (B6, folic acid (B9, and 4-aminobenzoic acid (B10. These analytes were separated optimizing the experimental conditions in 20 mM tetraborate buffer pH=9.2 as a BGE (background electrolyte, on a Beckman P/ACE System MDQ instrument, using uncoated fused silica capillary. The effective capillary length was of 49.5 cm, I.D.=50 μm, the applied voltage 20 kV and the temperature 25∘C. Detection was performed by a diode array detector at 214 nm for all vitamins except B5 (190 nm and B2a (260 nm. Separation time was about 9 min. After experimental conditions optimization, the proposed method was validated. Precision of migration time and corrected peak area, linearity range, LOD and LOQ, accuracy (recovery, robustness, and ruggedness were evaluated for each analyte demonstrating the good reliability of the method. Analyses of the pharmaceutical real samples were performed and confirmed the versatility of this method.

  3. Optimization and validation of a CZE method for rufloxacin hydrochloride determination in coated tablets.

    Science.gov (United States)

    Furlanetto, S; Orlandini, S; Porta, E La; Coran, S; Pinzauti, S

    2002-06-15

    A simple and rapid capillary electrophoresis method with UV detection was developed and validated for the determination of rufloxacin hydrochloride in coated tablets. An experimental design strategy (Doehlert design and desirability function) allowed the analytical parameters to be simultaneously optimized in order to determine rufloxacin hydrochloride with high peak area/migration time ratio, good efficiency and short analysis time. Optimized analyses were run using boric acid 0.10 M adjusted to pH 8.8 as BGE and setting voltage and temperature at 18 kV and 27 degrees C, respectively. Pefloxacin mesylate was used as internal standard and run time was about three minutes. The method was validated for the drug substance and the drug product according to the ICH3 guidelines. Robustness was tested by experimental design using an eight-run Plackett-Burman matrix.

  4. The CzeCOS Network

    Science.gov (United States)

    Havránková, Kateřina; Taufarová, Klára; Šigut, Ladislav; McGloin, Ryan; Acosta, Manuel; Dušek, Jiří; Krupková, Lenka; Macálková-Mžourková, Lenka; Pavelka, Marian; Dařenová, Eva; Yadav, Shilpi; Nguyen, Vinh; Guerra, Carlos; Janous, Dalibor; Marek, Michal V.

    2017-04-01

    The Global Change Research Institute of the Czech Academy of Sciences (CzechGlobe) have established a well-equipped network of ecosystem stations, with modern instrumentation for eco-physiological, plant physiological and micrometeorological studies, and estimation of GHG emissions. The network of stations (CzeCOS) covers the main terrestrial ecosystems of the Czech Republic (young and old coniferous forest, deciduous forest, mixed floodplain forest, grassland, wetland and cropland). The ecosystem stations are equipped with eddy covariance systems, soil and stem chamber systems for CO2 efflux and instruments for making micrometeorological measurements. The network enables detailed research to be conducted on topics such as: the carbon balance of different ecosystems, energy balance closure, the impact of current climate conditions on production and ecosystem disturbances during extreme weather conditions (drought, floods, winter storms, etc.) at regional, national and international scales. As a part of global networks (Fluxnet, ANAEe, ICOS), CzeCOS participates in evaluating and predicting environmental change and helps in the proposal of mitigation measures. Another important issue studied at some of the CzeCOS sites is the use of the eddy covariance method in sloping terrain in order to improve eddy covariance data processing for sites in this kind of terrain. Here we show specific results from the sites and outline the importance of the regional/national network for improving our knowledge about the exchange of matter and energy fluxes at different ecosystems. This study was supported by the Ministry of Education, Youth and Sports of CR within the National Sustainability Program I (NPU I), grant number LO1415 and LD 15040. Computational resources were provided by the CESNET LM2015042 and the CERIT Scientific Cloud LM2015085, provided under the programme "Projects of Large Research, Development, and Innovations Infrastructures".

  5. Rapid methods for determination of fluoxetine in pharmaceutical formulations.

    Science.gov (United States)

    Mandrioli, R; Pucci, V; Visini, D; Varani, G; Raggi, M A

    2002-08-01

    Two different analytical methods for the quality control of fluoxetine in commercial formulations have been developed and compared: a spectrofluorimetric method and a capillary zone electrophoretic (CZE) method. The fluorescence emission values were measured at lambda=293 nm when exciting at lambda=230 nm. The CZE method used an uncoated fused-silica capillary and pH 2.5 phosphate buffer as the background electrolyte. The extraction of fluoxetine from the capsules consisted of a simple one-step dissolution with methanol/water, filtration and dilution. Both methods gave satisfactory results in terms of precision; the best results were obtained for the electrophoretic method, with RSD% values always lower than 2.0%. The accuracy was assessed by means of recovery studies, which gave very good results, between 97.5 and 102.6%. Furthermore, both methods also have the advantage of being very rapid.

  6. Quantification of ascorbic acid and acetylsalicylic acid in effervescent tablets by CZE-UV and identification of related degradation products by heart-cut CZE-CZE-MS.

    Science.gov (United States)

    Neuberger, Sabine; Jooß, Kevin; Ressel, Christian; Neusüß, Christian

    2016-12-01

    Capillary electrophoresis is commonly applied for the analysis of pharmaceutical products due to its high separation efficiency and selectivity. For this purpose, electrospray-ionization-(ESI)-interfering additives or electrolytes are often required, which complicates the identification of impurities and degradation products by mass spectrometry (MS). Here, a capillary zone electrophoresis (CZE) method with ultraviolet (UV) absorption detection for the simultaneous determination and quantification of ascorbic acid and acetylsalicylic acid in effervescent tablets was developed. Related degradation products were identified via CZE-CZE-MS. Systematic optimization yielded 100 mM tricine (pH = 8.8) as appropriate background electrolyte, resulting in baseline separation of ascorbic acid, acetylsalicylic acid, and related anionic UV-active degradation products. The CZE-UV method was successfully validated regarding the guidelines of the Food and Drug Administration. The validated method was applied to trace the degradation rate of the active pharmaceutical ingredients at defined ambient conditions. A heart-cut CZE-CZE-MS approach, including a 4-port-nL-valve, was performed for the identification of the observed degradation products. This 2D setup enables a precise cutting of accurate sample volumes (20 nL) and the independent operation of two physically separated CZE dimensions, which is especially beneficial regarding MS detection. Hence, the ESI-interfering tricine electrolyte components were separated from the analytes in a second electrophoretic dimension prior to ESI-MS detection. The degradation products were identified as salicylic acid and mono- and diacetylated ascorbic acid. This setup is expected to be generally applicable for the mass spectrometric characterization of CZE separated analytes in highly ESI-interfering electrolyte systems. Graphical Abstract A CZE-UV method for the quantification of effervescent tablet ingredients and degradation products

  7. Characterization of cetuximab F{sub c/2} dimers by off-line CZE-MS

    Energy Technology Data Exchange (ETDEWEB)

    François, Yannis-Nicolas, E-mail: yfrancois@unistra.fr [Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), UDS-CNRS UMR 7140, Université de Strasbourg, Strasbourg (France); Biacchi, Michael; Said, Nassur; Renard, Charly [Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), UDS-CNRS UMR 7140, Université de Strasbourg, Strasbourg (France); Beck, Alain [Centre d' immunologie Pierre Fabre, Saint-Julien-en-Genevois (France); Gahoual, Rabah [Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), UDS-CNRS UMR 7140, Université de Strasbourg, Strasbourg (France); Laboratory of Bioanalytical Chemistry and Spectroscopy, Vrije Universiteit, Amsterdam (Netherlands); Leize-Wagner, Emmanuelle [Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), UDS-CNRS UMR 7140, Université de Strasbourg, Strasbourg (France)

    2016-02-18

    Monoclonal antibody (mAb) therapeutics attract the largest concern due to their strong therapeutic potency and specificity. The Fc region of mAbs is common to many new biotherapeutics as biosimilar, antibody drug conjugate or fusion protein. Fc region has consequences for Fc-mediated effector functions that might be desirable for therapeutic applications. As a consequence, there is a continuous need for improvement of analytical methods to enable fast and accurate characterization of biotherapeutics. Capillary zone electrophoresis-Mass spectrometry couplings (CZE-MS) appear really attractive methods for the characterization of biological samples. In this report, we used CZE-MS systems developed in house and native MS infusion to allow precise middle-up characterization of F{sub c/2} variant of cetuximab. Molecular weights were measured for three F{sub c/2} charge variants detected in the CZE separation of cetuximab subunits. Two F{sub c/2} C-terminal lysine variants were identified and separated. As the aim is to understand the presence of three peaks in the CZE separation for two F{sub c/2} subunits, we developed a strategy using CZE-UV/MALDI-MS and CZE-UV/ESI-MS to evaluate the role of N-glycosylation and C-terminal lysine truncation on the CZE separation. The chemical structure of N-glycosylation expressed on the Fc region of cetuximab does not influence CZE separation while C-terminal lysine is significantly influencing separation. In addition, native MS infusion demonstrated the characterization of F{sub c/2} dimers at pH 5.7 and 6.8 and the first separation of these dimers using CZE-MS. - Highlights: • We confirm the power of off-line CE/MS coupling for the separation of mAb isoforms. • We demonstrate the role of C-terminal lysine truncation in the separation process. • We demonstrate the presence of non-covalent interactions between Fc/2 subunit. • We identify Fc/2 homo- and heterodimers.

  8. Advanced portrayal of SMIL coating by allying CZE performance with in-capillary topographic and charge-related surface characterization

    Energy Technology Data Exchange (ETDEWEB)

    Stock, Lorenz G. [Division of Chemistry and Bioanalytics, University Salzburg, Hellbrunnerstrasse 34, 5020 Salzburg (Austria); Christian Doppler Laboratory for Innovative Tools for the Characterization of Biosimilars, Hellbrunnerstrasse 34, 5020 Salzburg (Austria); Leitner, Michael; Traxler, Lukas [Institute of Biophysics, Johannes Kepler University Linz, Gruberstrasse 40, 4020 Linz (Austria); Bonazza, Klaus [Institute of Chemical Technologies and Analytics, Vienna University of Technology, Getreidemarkt 9/164, 1060 Vienna (Austria); Leclercq, Laurent; Cottet, Hervé [Institut des Biomolécules Max Mousseron (IBMM), UMR 5247, CNRS, Université de Montpellier, Ecole Nationale Supérieure de Chimie de Montpellier, Place Eugène Bataillon, CC 1706, 34095 Montpellier (France); Friedbacher, Gernot [Institute of Chemical Technologies and Analytics, Vienna University of Technology, Getreidemarkt 9/164, 1060 Vienna (Austria); Ebner, Andreas [Institute of Biophysics, Johannes Kepler University Linz, Gruberstrasse 40, 4020 Linz (Austria); Stutz, Hanno, E-mail: hanno.stutz@sbg.ac.at [Division of Chemistry and Bioanalytics, University Salzburg, Hellbrunnerstrasse 34, 5020 Salzburg (Austria); Christian Doppler Laboratory for Innovative Tools for the Characterization of Biosimilars, Hellbrunnerstrasse 34, 5020 Salzburg (Austria)

    2017-01-25

    CZE performance is related to topography and charge distribution on the SMIL surface. • Topographic changes are confirmed by statistical methods.

  9. On-line solid phase extraction CZE for the simultaneous determination of lanthanum and gadolinium at picogram per liter levels.

    Science.gov (United States)

    Vizioli, Nora; Gil, Raúl; Martínez, Luis Dante; Silva, María Fernanda

    2009-08-01

    A non-specific on-line method is presented for the extraction and preconcentration of two rare earth elements using a microcartridge containing C(18)-derivatized silica particles prior to their analysis by CZE. The microcartridge, named analyte concentrator, was coupled on-line to the inlet of the separation capillary (fused-silica (FS) capillary, 75 microm id x12 cm from the inlet to the microcartidge and 37 cm from the microcartridge to the detector). The reversed-phase sorbent quantitatively retained gadolinium (Gd) and lanthanum (La) as 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol complexes in the presence of non-ionic micelles of polyethylene glycol tert-octylphenyl ether, enabling sample clean-up and concentration enhancement with minimum sample handling. The rare earth elements chelates were released from the sorbent with methanol and then analyzed by CZE with diode array detection. A background electrolyte of 20 mM sodium tetraborate containing 8% ACN, pH 9.0, was found to be optimal for the separation of metal chelates. The concentration limits of detection were lowered to picogram per liter levels (20 pg/L for La and 80 pg/L for Gd). A 1000-fold improvement in concentration sensitivity for La- and Gd-2-(5-bromo-2-pyridylazo)-5-diethylaminophenol complexes with respect to CZE without preconcentration was reached.

  10. Rapid methods for detection of bacteria

    DEFF Research Database (Denmark)

    Corfitzen, Charlotte B.; Andersen, B.Ø.; Miller, M.

    2006-01-01

    Traditional methods for detection of bacteria in drinking water e.g. Heterotrophic Plate Counts (HPC) or Most Probable Number (MNP) take 48-72 hours to give the result. New rapid methods for detection of bacteria are needed to protect the consumers against contaminations. Two rapid methods...

  11. Innovative rapid construction/reconstruction methods.

    Science.gov (United States)

    2005-07-01

    Innovative construction and reconstruction methods provide the opportunity to significantly reduce the time of roadway projects while maintaining the necessary quality of workmanship. The need for these rapid methods stems from the increase in ...

  12. A new method for rapid Canine retraction

    Directory of Open Access Journals (Sweden)

    "Khavari A

    2001-06-01

    Full Text Available Distraction osteogenesis method (Do in bone lengthening and rapid midpalatal expansion have shown the great ability of osteognic tissues for rapid bone formation under distraction force and special protocol with optimum rate of one millimeter per day. Periodontal membrane of teeth (PDM is the extension of periostium in the alveolar socked. Orthodontic force distracts PDM fibers in the tension side and then bone formation will begin.Objects: Rapid retraction of canine tooth into extraction space of first premolar by DO protocol in order to show the ability of the PDM in rapid bone formation. The other objective was reducing total orthodontic treatment time of extraction cases.Patients and Methods: Tweleve maxillary canines in six patients were retracted rapidly in three weeks by a custom-made tooth-born appliance. Radiographic records were taken to evaluate the effects of heavy applied force on canine and anchorage teeth.Results: Average retraction was 7.05 mm in three weeks (2.35 mm/week. Canines rotated distal- in by mean 3.5 degrees.Anchorage loss was from 0 to 0.8 mm with average of 0.3 mm.Root resorption of canines was negligible, and was not significant clinically. Periodontium was normal after rapid retraction. No hazard for pulp vitality was observed.Discussion: PDM responded well to heavy distraction force by Do protocol. Rapid canine retraction seems to be a safe method and can considerabely reduce orthodontic time.

  13. A scoping review of rapid review methods.

    Science.gov (United States)

    Tricco, Andrea C; Antony, Jesmin; Zarin, Wasifa; Strifler, Lisa; Ghassemi, Marco; Ivory, John; Perrier, Laure; Hutton, Brian; Moher, David; Straus, Sharon E

    2015-09-16

    Rapid reviews are a form of knowledge synthesis in which components of the systematic review process are simplified or omitted to produce information in a timely manner. Although numerous centers are conducting rapid reviews internationally, few studies have examined the methodological characteristics of rapid reviews. We aimed to examine articles, books, and reports that evaluated, compared, used or described rapid reviews or methods through a scoping review. MEDLINE, EMBASE, the Cochrane Library, internet websites of rapid review producers, and reference lists were searched to identify articles for inclusion. Two reviewers independently screened literature search results and abstracted data from included studies. Descriptive analysis was conducted. We included 100 articles plus one companion report that were published between 1997 and 2013. The studies were categorized as 84 application papers, seven development papers, six impact papers, and four comparison papers (one was included in two categories). The rapid reviews were conducted between 1 and 12 months, predominantly in Europe (58 %) and North America (20 %). The included studies failed to report 6 % to 73 % of the specific systematic review steps examined. Fifty unique rapid review methods were identified; 16 methods occurred more than once. Streamlined methods that were used in the 82 rapid reviews included limiting the literature search to published literature (24 %) or one database (2 %), limiting inclusion criteria by date (68 %) or language (49 %), having one person screen and another verify or screen excluded studies (6 %), having one person abstract data and another verify (23 %), not conducting risk of bias/quality appraisal (7 %) or having only one reviewer conduct the quality appraisal (7 %), and presenting results as a narrative summary (78 %). Four case studies were identified that compared the results of rapid reviews to systematic reviews. Three studies found that the conclusions between

  14. Rapid determination of capsaicinoids by colorimetric method

    OpenAIRE

    Ryu, Wang-Kyun; Kim, Hee-Woong; Kim, Geun-Dong; Rhee, Hae-Ik

    2016-01-01

    Capsaicinoids, the pungent component of chili peppers, are generally analyzed by precise analytical techniques, such as gas chromatography and high-performance liquid chromatography (HPLC), but these are not practical for the mass analyses of samples. To analyze mass samples rapidly, a colorimetric method was suggested. In this work, pigments and capsaicinoids were efficiently separated from chili pepper extract by sequential solid–liquid extraction and liquid–liquid extraction in test tubes ...

  15. Rapid Column Extraction method for SoilRapid Column Extraction method for Soil

    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, Sherrod, L. III; Culligan, Brian K.

    2005-11-07

    The analysis of actinides in environmental soil and sediment samples is very important for environmental monitoring as well as for emergency preparedness. A new, rapid actinide separation method has been developed and implemented that provides total dissolution of large soil samples, high chemical recoveries and effective removal of matrix interferences. This method uses stacked TEVA Resin{reg_sign}, TRU Resin{reg_sign} and DGA-Resin{reg_sign} cartridges from Eichrom Technologies (Darien, IL, USA) that allows the rapid separation of plutonium (Pu) neptunium (Np), uranium (U), americium (Am), and curium (Cm) using a single multi-stage column combined with alpha spectrometry. The method combines a rapid fusion step for total dissolution to dissolve refractory analytes and matrix removal using cerium fluoride precipitation to remove the difficult soil matrix. By using vacuum box cartridge technology with rapid flow rates, sample preparation time is minimized.

  16. Rapid determination of capsaicinoids by colorimetric method.

    Science.gov (United States)

    Ryu, Wang-Kyun; Kim, Hee-Woong; Kim, Geun-Dong; Rhee, Hae-Ik

    2017-10-01

    Capsaicinoids, the pungent component of chili peppers, are generally analyzed by precise analytical techniques, such as gas chromatography and high-performance liquid chromatography (HPLC), but these are not practical for the mass analyses of samples. To analyze mass samples rapidly, a colorimetric method was suggested. In this work, pigments and capsaicinoids were efficiently separated from chili pepper extract by sequential solid-liquid extraction and liquid-liquid extraction in test tubes followed by a colorimetric analysis on the capsaicinoids by a selective chromogenic reaction with Gibbs reagent (2,6-dichloroquinone-4-chloroimide). In the comparison of the capsaicinoid content by the colorimetric method and HPLC using acetone extracts of fresh pepper and dry red pepper as samples, R2 was 0.9973 and 0.9816, respectively, which shows a high linear correlation. In addition, a minimum of 1 μg/mL capsaicinoids can be detected and it was therefore determined that the method can efficiently analyze a great quantity of samples in a short time. Copyright © 2016. Published by Elsevier B.V.

  17. Rapid determination of capsaicinoids by colorimetric method

    Directory of Open Access Journals (Sweden)

    Wang-Kyun Ryu

    2017-10-01

    Full Text Available Capsaicinoids, the pungent component of chili peppers, are generally analyzed by precise analytical techniques, such as gas chromatography and high-performance liquid chromatography (HPLC, but these are not practical for the mass analyses of samples. To analyze mass samples rapidly, a colorimetric method was suggested. In this work, pigments and capsaicinoids were efficiently separated from chili pepper extract by sequential solid–liquid extraction and liquid–liquid extraction in test tubes followed by a colorimetric analysis on the capsaicinoids by a selective chromogenic reaction with Gibbs reagent (2,6-dichloroquinone-4-chloroimide. In the comparison of the capsaicinoid content by the colorimetric method and HPLC using acetone extracts of fresh pepper and dry red pepper as samples, R2 was 0.9973 and 0.9816, respectively, which shows a high linear correlation. In addition, a minimum of 1 μg/mL capsaicinoids can be detected and it was therefore determined that the method can efficiently analyze a great quantity of samples in a short time.

  18. Determination of endocrine disruptors in honey by CZE-MS using restricted access materials for matrix cleanup.

    Science.gov (United States)

    Rodríguez-Gonzalo, Encarnación; Domínguez-Alvarez, Javier; García-Gómez, Diego; García-Jiménez, María-Guadalupe; Carabias-Martínez, Rita

    2010-07-01

    An analytical method based on CZE coupled to ESI-MS is proposed for the identification and simultaneous quantification of several endocrine-disrupting chemicals in honey. The target compounds were the chlorophenols: 2,4-dichlorophenol, 2,4,5-trichlorophenol and pentachlorophenol, and bisphenol-A, 4-tert-butylphenol, and 4-tert-butylbenzoic acid. A two-step optimization of the ESI-MS detection was carried out. First, the organic solvent present in the sheath liquid was selected and its effect on the analytical signal was studied. The best results in terms of the intensity of the MS signals were obtained with methanol. Thus, an experimental design technique (Doehlert type) was used for the optimization of the other parameters: the NH(3) concentration in the sheath liquid, the flow of the sheath liquid, the nebulizer pressure in ESI, and the drying gas temperature and flow. Here, we developed a new sample treatment based on the combined use of a restricted access material and a polymeric sorbent for SPE. The LOD achieved were in the range of 5-31 ng/g. The intraday precision of the proposed method was determined from replicate analyses (n=4) at a concentration level of 50 ng/g, with RSD values in the range of 15-23%. The results revealed that the proposed method is suitable for the reliable quantification of endocrine-disrupting chemicals in honey at nanograms per gram levels.

  19. Rapid method for detection of salmonella in meat

    DEFF Research Database (Denmark)

    2016-01-01

    The present invention relates to a rapid method for the detection of Salmonella in meat as well as to a kit for performing said method. The method provides a time-to-result of less than 8 hours.......The present invention relates to a rapid method for the detection of Salmonella in meat as well as to a kit for performing said method. The method provides a time-to-result of less than 8 hours....

  20. Methods and compositions for rapid thermal cycling

    Science.gov (United States)

    Beer, Neil Reginald; Benett, William J.; Frank, James M.; Deotte, Joshua R.; Spadaccini, Christopher

    2015-10-27

    The rapid thermal cycling of a material is targeted. A microfluidic heat exchanger with an internal porous medium is coupled to tanks containing cold fluid and hot fluid. Fluid flows alternately from the cold tank and the hot tank into the porous medium, cooling and heating samples contained in the microfluidic heat exchanger's sample wells. A valve may be coupled to the tanks and a pump, and switching the position of the valve may switch the source and direction of fluid flowing through the porous medium. A controller may control the switching of valve positions based on the temperature of the samples and determined temperature thresholds. A sample tray for containing samples to be thermally cycled may be used in conjunction with the thermal cycling system. A surface or internal electrical heater may aid in heating the samples, or may replace the necessity for the hot tank.

  1. Hydration, charge, size, and shape characteristics of peptides from their CZE analyses.

    Science.gov (United States)

    Peirotti, Marta B; Piaggio, Maria V; Deiber, Julio A

    2008-02-01

    A CZE model is presented for peptide characterization on the basis of well-established physicochemical equations. The effective mobility is used as basic data in the model to estimate relevant peptide properties such as, for instance, hydration, net and total electrical charge numbers, hydrodynamic size and shape, particle average orientation, and pH-microenvironment from the charge regulation phenomenon. Therefore 102 experimental effective mobilities of different peptides are studied and discussed in relation to previous work. An equation for the estimation of peptide hydration as a function of ionizing, polar, and non-polar amino acid residues is included in the model. It is also shown that the shape-orientation factor of peptides may be either lower or higher than one, and its value depends on a complex interplay among total charge number, molar mass, hydration, and amino acid sequence.

  2. RAPID METHOD FOR DETERMINATION OF RADIOSTRONTIUM IN EMERGENCY MILK SAMPLES

    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, S.; Culligan, B.

    2008-07-17

    A new rapid separation method for radiostrontium in emergency milk samples was developed at the Savannah River Site (SRS) Environmental Bioassay Laboratory (Aiken, SC, USA) that will allow rapid separation and measurement of Sr-90 within 8 hours. The new method uses calcium phosphate precipitation, nitric acid dissolution of the precipitate to coagulate residual fat/proteins and a rapid strontium separation using Sr Resin (Eichrom Technologies, Darien, IL, USA) with vacuum-assisted flow rates. The method is much faster than previous method that use calcination or cation exchange pretreatment, has excellent chemical recovery, and effectively removes beta interferences. When a 100 ml sample aliquot is used, the method has a detection limit of 0.5 Bq/L, well below generic emergency action levels.

  3. A universal, rapid, and inexpensive method for genomic DNA ...

    Indian Academy of Sciences (India)

    MOHAMMED BAQUR SAHIB A. AL-SHUHAIB

    Abstract. There is no 'one' procedure for extracting DNA from the whole blood of both mammals and birds, since each species has a unique property that require different methods to release its own DNA. Therefore, to obtain genomic DNA, a universal, rapid, and noncostly method was developed. A very simple biological ...

  4. Rapid, cost-effective liquid chromatograghic method for the ...

    African Journals Online (AJOL)

    GRACE

    2006-07-03

    Jul 3, 2006 ... 1Department of Medicinal Chemistry and Quality Control, National Institute for Pharmaceutical Research and Development, Abuja,. Nigeria. ... A rapid and cost effective method for the analysis of metronidazole in biological samples was ... effective HPLC method of assaying metronidazole both in.

  5. A universal, rapid, and inexpensive method for genomic DNA ...

    Indian Academy of Sciences (India)

    ... of both mammals and birds, since each species has a unique property that require different methods to release its own DNA. Therefore, to obtain genomic DNA, a universal, rapid, and noncostly method was developed. A very simple biological basis is followed in this procedure, in which, when the bloodis placed in water, ...

  6. Methods for Rapid Screening in Woody Plant Herbicide Development

    Directory of Open Access Journals (Sweden)

    William Stanley

    2014-07-01

    Full Text Available Methods for woody plant herbicide screening were assayed with the goal of reducing resources and time required to conduct preliminary screenings for new products. Rapid screening methods tested included greenhouse seedling screening, germinal screening, and seed screening. Triclopyr and eight experimental herbicides from Dow AgroSciences (DAS 313, 402, 534, 548, 602, 729, 779, and 896 were tested on black locust, loblolly pine, red maple, sweetgum, and water oak. Screening results detected differences in herbicide and species in all experiments in much less time (days to weeks than traditional field screenings and consumed significantly less resources (<500 mg acid equivalent per herbicide per screening. Using regression analysis, various rapid screening methods were linked into a system capable of rapidly and inexpensively assessing herbicide efficacy and spectrum of activity. Implementation of such a system could streamline early-stage herbicide development leading to field trials, potentially freeing resources for use in development of beneficial new herbicide products.

  7. Methods and systems for rapid prototyping of high density circuits

    Science.gov (United States)

    Palmer, Jeremy A [Albuquerque, NM; Davis, Donald W [Albuquerque, NM; Chavez, Bart D [Albuquerque, NM; Gallegos, Phillip L [Albuquerque, NM; Wicker, Ryan B [El Paso, TX; Medina, Francisco R [El Paso, TX

    2008-09-02

    A preferred embodiment provides, for example, a system and method of integrating fluid media dispensing technology such as direct-write (DW) technologies with rapid prototyping (RP) technologies such as stereolithography (SL) to provide increased micro-fabrication and micro-stereolithography. A preferred embodiment of the present invention also provides, for example, a system and method for Rapid Prototyping High Density Circuit (RPHDC) manufacturing of solderless connectors and pilot devices with terminal geometries that are compatible with DW mechanisms and reduce contact resistance where the electrical system is encapsulated within structural members and manual electrical connections are eliminated in favor of automated DW traces. A preferred embodiment further provides, for example, a method of rapid prototyping comprising: fabricating a part layer using stereolithography and depositing thermally curable media onto the part layer using a fluid dispensing apparatus.

  8. CZE/PAD and HPLC-UV/PAD Profile of Flavonoids from Maytenus aquifolium and Maytenus ilicifolia “espinheira santa” Leaves Extracts

    Directory of Open Access Journals (Sweden)

    Cristina A. Diagone

    2012-01-01

    Full Text Available This paper describes the application of HPLC and CZE to analyze flavonoids in the leaves of Maytenus ilicifolia and Maytenus aquifolium, which are species widely used in Brazilian folk medicine. The two species showed different flavonoid profiles, but acidic hydrolysis of the Maytenus extracts confirmed that all these compounds are quercetin or kaempferol derivatives. A comparison of the CZE and HPLC profiles of Maytenus extracts showed numerous flavonoid peaks using HPLC. However, the advantages of CZE such as analysis without requiring clean-up and less generation of chemical waste than with HPLC point to the potential of the CZE technique for the quality control (routine analysis of “espinheira santa” phytopharmaceuticals.

  9. A rapid ultrasound particle agglutination method for HIV antibody detection: Comparison with conventional rapid HIV tests.

    Science.gov (United States)

    Bystryak, Simon; Ossina, Natalya

    2017-11-01

    We present the results of the feasibility and preliminary studies on analytical performance of a rapid test for detection of human immunodeficiency virus (HIV) antibodies in human serum or plasma that is an important advance in detecting HIV infection. Current methods for rapid testing of antibodies against HIV are qualitative and exhibit poor sensitivity (limit of detection). In this paper, we describe an ultrasound particle agglutination (UPA) method that leads to a significant increase of the sensitivity of conventional latex agglutination tests for HIV antibody detection in human serum or plasma. The UPA method is based on the use of: 1) a dual mode ultrasound, wherein a first single-frequency mode is used to accelerate the latex agglutination process, and then a second swept-frequency mode of sonication is used to disintegrate non-specifically bound aggregates; and 2) a numerical assessment of results of the agglutination process. The numerical assessment is carried out by optical detection and analysis of moving patterns in the resonator cell during the swept-frequency mode. The single-step UPA method is rapid and more sensitive than the three commercial rapid HIV test kits analyzed in the study: analytical sensitivity of the new UPA method was found to be 510-, 115-, and 80-fold higher than that for Capillus™, Multispot™ and Uni-Gold™ Recombigen HIV antibody rapid test kits, respectively. The newly developed UPA method opens up additional possibilities for detection of a number of clinically significant markers in point-of-care settings. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. CZE/PAD and HPLC-UV/PAD Profile of Flavonoids from Maytenus aquifolium and Maytenus ilicifolia “espinheira santa” Leaves Extracts

    OpenAIRE

    Cristina A. Diagone; Renata Colombo; Lanças, Fernando M.; Yariwake, Janete H.

    2012-01-01

    This paper describes the application of HPLC and CZE to analyze flavonoids in the leaves of Maytenus ilicifolia and Maytenus aquifolium, which are species widely used in Brazilian folk medicine. The two species showed different flavonoid profiles, but acidic hydrolysis of the Maytenus extracts confirmed that all these compounds are quercetin or kaempferol derivatives. A comparison of the CZE and HPLC profiles of Maytenus extracts showed numerous flavonoid peaks using HPLC. However, the advant...

  11. RAPID SEPARATION METHOD FOR ACTINIDES IN EMERGENCY SOIL SAMPLES

    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, S.; Culligan, B.; Noyes, G.

    2009-11-09

    A new rapid method for the determination of actinides in soil and sediment samples has been developed at the Savannah River Site Environmental Lab (Aiken, SC, USA) that can be used for samples up to 2 grams in emergency response situations. The actinides in soil method utilizes a rapid sodium hydroxide fusion method, a lanthanum fluoride soil matrix removal step, and a streamlined column separation process with stacked TEVA, TRU and DGA Resin cartridges. Lanthanum was separated rapidly and effectively from Am and Cm on DGA Resin. Vacuum box technology and rapid flow rates are used to reduce analytical time. Alpha sources are prepared using cerium fluoride microprecipitation for counting by alpha spectrometry. The method showed high chemical recoveries and effective removal of interferences. This new procedure was applied to emergency soil samples received in the NRIP Emergency Response exercise administered by the National Institute for Standards and Technology (NIST) in April, 2009. The actinides in soil results were reported within 4-5 hours with excellent quality.

  12. Absolute quantitation of host cell proteins in recombinant human monoclonal antibodies with an automated CZE-ESI-MS/MS system.

    Science.gov (United States)

    Zhu, Guijie; Sun, Liangliang; Linkous, Travis; Kernaghan, Dawn; McGivney, James B; Dovichi, Norman J

    2014-05-01

    We report the first use of CZE for absolute characterization of host cell proteins (HCPs) in recombinant human monoclonal antibodies. An electrokinetically pumped nanoelectrospray interface was used to couple CZE with a tandem mass spectrometer. Three isotopic-labeled peptides (LSFDKDAMVAR, VDIVENQAMDTR, and LVSDEMVVELIEK) were synthesized by direct incorporation of an isotope-labeled lysine or arginine. The heavy-labeled peptides were spiked in the HCP digests at known concentrations. After CZE-ESI-MS/MS analysis, the peaks of native and isotopic-labeled peptides were extracted with mass tolerance ≤ 5 ppm from the electropherograms, and the ratios of peak area between native and isotopic-labeled peptides pairs were calculated. Calibration curves (the ratios of peak area versus spiked peptide amount) with R(2) values of 0.999, 0.997, and 0.999 were obtained for the three HCP peptides, and the absolute amounts of the three proteins present were determined to be at the picomole level in a 20 μg sample of digested HCPs. The target proteins were present at the 7-30 ppt level in the purified HCP samples. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Rapid method to estimate temperature changes in electronics elements

    Directory of Open Access Journals (Sweden)

    Oborskii G. A., Savel’eva O. S., Shikhireva Yu. V.

    2014-06-01

    Full Text Available Thermal behavior of electronic equipment is the determining factor for performing rapid assessment of the effectiveness of design and operation of the equipment. The assessment method proposed in this article consists in fixation of an infrared video stream from the surface of the device and converting it into a visible flow by means of a thermal imager, splitting it into component colors and their further processing using parabolic transformation. The result of the transformation is the number used as a rapid criterion for estimation of distribution stability of heat in the equipment.

  14. Rapid methods for detection of bacterial resistance to antibiotics.

    Science.gov (United States)

    March-Rosselló, Gabriel Alberto

    2017-03-01

    The most widely used antibiotic susceptibility testing methods in Clinical Microbiology are based on the phenotypic detection of antibiotic resistance by measuring bacterial growth in the presence of the antibiotic being tested. These conventional methods take typically 24hours to obtain results. Here we review the main techniques for rapid determination of antibiotic susceptibility. Data obtained with different methods such as molecular techniques, microarrays, commercial methods used in work routine, immunochromatographic methods, colorimetric methods, image methods, nephelometry, MALDI-TOF mass spectrometry, flow cytometry, chemiluminescence and bioluminescence, microfluids and methods based on cell disruption are analysed in detail. Copyright © 2016 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  15. Evaluation of a Rapid Method of Determination of Plasma Fibrinogen

    Science.gov (United States)

    Thomson, G. W.; McSherry, B. J.; Valli, V. E. O.

    1974-01-01

    An evaluation was made of a rapid semiautomated method of determining fibrinogen levels in bovine plasma. This method, the fibrometer method of Morse, Panek and Menga (8), is based on the principle that when thrombin is added to suitably diluted plasma the time of clotting is linearly related to the fibrinogen concentration. A standard curve prepared using bovine plasma had an r value of .9987 and analysis of variance showed there was no significant deviation from regression. A comparison of the fibrometer method and the biuret method of Ware, Guest and Seegers done on 158 bovine plasma samples showed good correlation between the two methods. It was concluded that the fibrometer method does measure bovine fibrinogen and has considerable merit for use in clinical diseases of cattle. PMID:4277474

  16. A simple and rapid method for determining transgenic cotton plants.

    Science.gov (United States)

    Zhang, Baohong; Wang, Hongmei; Liu, Fang; Wang, Qinglian

    2013-01-01

    Determining transgenic events is a critical step for obtaining transgenic plants as well as the later stage of application. Traditional methods, such as Northern blotting and qRT-PCR, for determining transgenic events either require radioactively labeled substrates, expensive instruments, or long-time commitments, which result in lab and time-consuming as well as expensive costs. These methods also require destroying the transgenic events. In this chapter, we present a simple and rapid method for determining transgenic cotton plants in both laboratory and field conditions. This method is based on the sensitivity of transgenic and non-transgenic plants to a specific chemical, such as antibiotics or herbicides. This method will facilitate the screening of transgenic events, save time, reduce cost, and speed up the application of transgenic technology on cotton breeding and production. More important, this is a nondestructive bioassay method; the transgenic plants can be transferred into greenhouse or field for the later study after the detection process.

  17. Rapid and selective method for quantitation of metronidazole in pharmaceuticals.

    Science.gov (United States)

    Sanyal, A K

    1988-01-01

    A selective and highly sensitive assay for N-1-substituted nitroimidazoles has been modified and adapted for rapid estimation of metronidazole in pharmaceuticals. The color reaction is based on diazotization of sulfanilamide with the nitrite ions liberated by alkaline hydrolysis of metronidazole and subsequent coupling of the diazonium salt with N-1-(naphthyl)-ethylenediamine dihydrochloride. This method is applicable for the assay of benzoyl metronidazole in oral suspension. Officially recommended excipients and preservatives do not interfere.

  18. Preparing Silica Aerogel Monoliths via a Rapid Supercritical Extraction Method

    Science.gov (United States)

    Gorka, Caroline A.

    2014-01-01

    A procedure for the fabrication of monolithic silica aerogels in eight hours or less via a rapid supercritical extraction process is described. The procedure requires 15-20 min of preparation time, during which a liquid precursor mixture is prepared and poured into wells of a metal mold that is placed between the platens of a hydraulic hot press, followed by several hours of processing within the hot press. The precursor solution consists of a 1.0:12.0:3.6:3.5 x 10-3 molar ratio of tetramethylorthosilicate (TMOS):methanol:water:ammonia. In each well of the mold, a porous silica sol-gel matrix forms. As the temperature of the mold and its contents is increased, the pressure within the mold rises. After the temperature/pressure conditions surpass the supercritical point for the solvent within the pores of the matrix (in this case, a methanol/water mixture), the supercritical fluid is released, and monolithic aerogel remains within the wells of the mold. With the mold used in this procedure, cylindrical monoliths of 2.2 cm diameter and 1.9 cm height are produced. Aerogels formed by this rapid method have comparable properties (low bulk and skeletal density, high surface area, mesoporous morphology) to those prepared by other methods that involve either additional reaction steps or solvent extractions (lengthier processes that generate more chemical waste).The rapid supercritical extraction method can also be applied to the fabrication of aerogels based on other precursor recipes. PMID:24637334

  19. Rapid Enzymatic Method for Pectin Methyl Esters Determination

    Directory of Open Access Journals (Sweden)

    Lucyna Łękawska-Andrinopoulou

    2013-01-01

    Full Text Available Pectin is a natural polysaccharide used in food and pharma industries. Pectin degree of methylation is an important parameter having significant influence on pectin applications. A rapid, fully automated, kinetic flow method for determination of pectin methyl esters has been developed. The method is based on a lab-made analyzer using the reverse flow-injection/stopped flow principle. Methanol is released from pectin by pectin methylesterase in the first mixing coil. Enzyme working solution is injected further downstream and it is mixed with pectin/pectin methylesterase stream in the second mixing coil. Methanol is oxidized by alcohol oxidase releasing formaldehyde and hydrogen peroxide. This reaction is coupled to horse radish peroxidase catalyzed reaction, which gives the colored product 4-N-(p-benzoquinoneimine-antipyrine. Reaction rate is proportional to methanol concentration and it is followed using Ocean Optics USB 2000+ spectrophotometer. The analyzer is fully regulated by a lab written LabVIEW program. The detection limit was 1.47 mM with an analysis rate of 7 samples h−1. A paired t-test with results from manual method showed that the automated method results are equivalent to the manual method at the 95% confidence interval. The developed method is rapid and sustainable and it is the first application of flow analysis in pectin analysis.

  20. Numerical Non-Equilibrium and Smoothing of Solutions in The Difference Method for Plane 2-Dimensional Adhesive Joints / Nierównowaga Numeryczna i Wygładzanie Rozwiazań w Metodzie Różnicowej Dla Dwuwymiarowych Połączeń Klejowych

    Directory of Open Access Journals (Sweden)

    Rapp Piotr

    2016-03-01

    Full Text Available The subject of the paper is related to problems with numerical errors in the finite difference method used to solve equations of the theory of elasticity describing 2- dimensional adhesive joints in the plane stress state. Adhesive joints are described in terms of displacements by four elliptic partial differential equations of the second order with static and kinematic boundary conditions. If adhesive joint is constrained as a statically determinate body and is loaded by a self-equilibrated loading, the finite difference solution is sensitive to kinematic boundary conditions. Displacements computed at the constraints are not exactly zero. Thus, the solution features a numerical error as if the adhesive joint was not in equilibrium. Herein this phenomenon is called numerical non-equilibrium. The disturbances in displacements and stress distributions can be decreased or eliminated by a correction of loading acting on the adhesive joint or by smoothing of solutions based on Dirichlet boundary value problem.

  1. SIMS: a hybrid method for rapid conformational analysis.

    Directory of Open Access Journals (Sweden)

    Bryant Gipson

    Full Text Available Proteins are at the root of many biological functions, often performing complex tasks as the result of large changes in their structure. Describing the exact details of these conformational changes, however, remains a central challenge for computational biology due the enormous computational requirements of the problem. This has engendered the development of a rich variety of useful methods designed to answer specific questions at different levels of spatial, temporal, and energetic resolution. These methods fall largely into two classes: physically accurate, but computationally demanding methods and fast, approximate methods. We introduce here a new hybrid modeling tool, the Structured Intuitive Move Selector (sims, designed to bridge the divide between these two classes, while allowing the benefits of both to be seamlessly integrated into a single framework. This is achieved by applying a modern motion planning algorithm, borrowed from the field of robotics, in tandem with a well-established protein modeling library. sims can combine precise energy calculations with approximate or specialized conformational sampling routines to produce rapid, yet accurate, analysis of the large-scale conformational variability of protein systems. Several key advancements are shown, including the abstract use of generically defined moves (conformational sampling methods and an expansive probabilistic conformational exploration. We present three example problems that sims is applied to and demonstrate a rapid solution for each. These include the automatic determination of "active" residues for the hinge-based system Cyanovirin-N, exploring conformational changes involving long-range coordinated motion between non-sequential residues in Ribose-Binding Protein, and the rapid discovery of a transient conformational state of Maltose-Binding Protein, previously only determined by Molecular Dynamics. For all cases we provide energetic validations using well

  2. A novel method for rapid in vitro radiobioassay

    Science.gov (United States)

    Crawford, Evan Bogert

    Rapid and accurate analysis of internal human exposure to radionuclides is essential to the effective triage and treatment of citizens who have possibly been exposed to radioactive materials in the environment. The two most likely scenarios in which a large number of citizens would be exposed are the detonation of a radiation dispersal device (RDD, "dirty bomb") or the accidental release of an isotope from an industrial source such as a radioisotopic thermal generator (RTG). In the event of the release and dispersion of radioactive materials into the environment in a large city, the entire population of the city -- including all commuting workers and tourists -- would have to be rapidly tested, both to satisfy the psychological needs of the citizens who were exposed to the mental trauma of a possible radiation dose, and to satisfy the immediate medical needs of those who received the highest doses and greatest levels of internal contamination -- those who would best benefit from rapid, intensive medical care. In this research a prototype rapid screening method to screen urine samples for the presence of up to five isotopes, both individually and in a mixture, has been developed. The isotopes used to develop this method are Co-60, Sr-90, Cs-137, Pu-238, and Am-241. This method avoids time-intensive chemical separations via the preparation and counting of a single sample on multiple detectors, and analyzing the spectra for isotope-specific markers. A rapid liquid-liquid separation using an organic extractive scintillator can be used to help quantify the activity of the alpha-emitting isotopes. The method provides quantifiable results in less than five minutes for the activity of beta/gamma-emitting isotopes when present in the sample at the intervention level as defined by the Centers for Disease Control and Prevention (CDC), and quantifiable results for the activity levels of alpha-emitting isotopes present at their respective intervention levels in approximately 30

  3. Rapid assessment methods in eye care: An overview

    Directory of Open Access Journals (Sweden)

    Srinivas Marmamula

    2012-01-01

    Full Text Available Reliable information is required for the planning and management of eye care services. While classical research methods provide reliable estimates, they are prohibitively expensive and resource intensive. Rapid assessment (RA methods are indispensable tools in situations where data are needed quickly and where time- or cost-related factors prohibit the use of classical epidemiological surveys. These methods have been developed and field tested, and can be applied across almost the entire gamut of health care. The 1990s witnessed the emergence of RA methods in eye care for cataract, onchocerciasis, and trachoma and, more recently, the main causes of avoidable blindness and visual impairment. The important features of RA methods include the use of local resources, simplified sampling methodology, and a simple examination protocol/data collection method that can be performed by locally available personnel. The analysis is quick and easy to interpret. The entire process is inexpensive, so the survey may be repeated once every 5-10 years to assess the changing trends in disease burden. RA survey methods are typically linked with an intervention. This article provides an overview of the RA methods commonly used in eye care, and emphasizes the selection of appropriate methods based on the local need and context.

  4. A rapid DNA extraction method suitable for human papillomavirus detection.

    Science.gov (United States)

    Brestovac, Brian; Wong, Michelle E; Costantino, Paul S; Groth, David

    2014-04-01

    Infection with oncogenic human papillomavirus (HPV) genotypes is necessary for the development of cervical cancer. Testing for HPV DNA from liquid based cervical samples can be used as an adjunct to traditional cytological screening. In addition there are ongoing viral load, genotyping, and prevalence studies. Therefore, a sensitive DNA extraction method is needed to maximize the efficiency of HPV DNA detection. The XytXtract Tissue kit is a DNA extraction kit that is rapid and so could be useful for HPV testing, particularly in screening protocols. This study was undertaken to determine the suitability of this method for HPV detection. DNA extraction from HeLa and Caski cell lines containing HPV 18 and 16 respectively together with DNA from five liquid based cervical samples were used in a HPV PCR assay. DNA was also extracted using the QIAamp DNA mini kit (Qiagen, Hilden, Germany) as a comparison. DNA extracts were serially diluted and assayed. HPV DNA was successfully detected in cell lines and cervical samples using the XytXtract Tissue kit. In addition, the XytXtract method was found to be more sensitive than the QIAmp method as determined by a dilution series of the extracted DNA. While the XytXtract method is a closed, the QIAamp method uses a spin column with possible loss of DNA through DNA binding competition of the matrix, which could impact on the final extraction efficiency. The XytXtract is a cheap, rapid and efficient method for extracting HPV DNA from both cell lines and liquid based cervical samples. © 2014 Wiley Periodicals, Inc.

  5. Advanced portrayal of SMIL coating by allying CZE performance with in-capillary topographic and charge-related surface characterization.

    Science.gov (United States)

    Stock, Lorenz G; Leitner, Michael; Traxler, Lukas; Bonazza, Klaus; Leclercq, Laurent; Cottet, Hervé; Friedbacher, Gernot; Ebner, Andreas; Stutz, Hanno

    2017-01-25

    A successive multiple ionic polymer layer (SMIL) coating composed of four layers improved the capillary electrophoretic separation of a recombinant major birch pollen allergen and closely related variants when poly(acrylamide-co-2-acrylamido-2-methyl-1-propansulfonate) (55% PAMAMPS) replaced dextran sulfate as terminal SMIL layer. 55% PAMAMPS decelerated the electroosmotic flow (EOF) due to its lower charge density. Atomic force microscopy (AFM) was used to investigate SMIL properties directly on the inner capillary surface and to relate them to EOF measurements and results of associated CZE separations of a mixture of model proteins and peptides that were performed in the same capillary. For the first time, AFM-based biosensing topography and recognition imaging mode (TREC) under liquid conditions was applied for a sequential characterization of the inner surface of a SMIL coated capillary after selected treatments including pristine SMIL, SMIL after contact with the model mixture, after alkaline rinsing, and the replenishment of the terminal polyelectrolyte layer. A cantilever with tip-tethered avidin was used to determine the charge homogeneity of the SMIL surface in the TREC mode. SMIL coated rectangular capillaries with 100 μm internal diameter assured accessibility of the inner surface for this cantilever type. Observed changes in CZE performance and EOF mobility during capillary treatment were also reflected by alterations in surface roughness and charge distribution of the SMIL coating. A renewal of the terminal SMIL layer restored the original surface properties of SMIL and the separation performance. The alliance of the novel TREC approach and CZE results allows for an improved understanding and a comprehensive insight in effects occurring on capillary coatings. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Radiometric method for the rapid detection of Leptospira organisms

    Energy Technology Data Exchange (ETDEWEB)

    Manca, N.; Verardi, R.; Colombrita, D.; Ravizzola, G.; Savoldi, E.; Turano, A.

    1986-02-01

    A rapid and sensitive radiometric method for detection of Leptospira interrogans serovar pomona and Leptospira interrogans serovar copenhageni is described. Stuart's medium and Middlebrook TB (12A) medium supplemented with bovine serum albumin, catalase, and casein hydrolysate and labeled with /sup 14/C-fatty acids were used. The radioactivity was measured in a BACTEC 460. With this system, Leptospira organisms were detected in human blood in 2 to 5 days, a notably shorter time period than that required for the majority of detection techniques.

  7. Method for rapidly determining a pulp kappa number using spectrophotometry

    Science.gov (United States)

    Chai, Xin-Sheng; Zhu, Jun Yong

    2002-01-01

    A system and method for rapidly determining the pulp kappa number through direct measurement of the potassium permanganate concentration in a pulp-permanganate solution using spectrophotometry. Specifically, the present invention uses strong acidification to carry out the pulp-permanganate oxidation reaction in the pulp-permanganate solution to prevent the precipitation of manganese dioxide (MnO.sub.2). Consequently, spectral interference from the precipitated MnO.sub.2 is eliminated and the oxidation reaction becomes dominant. The spectral intensity of the oxidation reaction is then analyzed to determine the pulp kappa number.

  8. Rapid coulometric method for the Kjeldahl determination of nitrogen.

    Science.gov (United States)

    Boström, C A; Cedergren, A; Johansson, G; Pettersson, I

    1974-11-01

    A rapid coulometric method for the Kjeldahl determination of nitrogen is described. The samples are digested by means of the Tecator AB digestion system which permits forty samples to be digested at the same time. The digestion products are diluted to 75 ml and 1 ml is coulometrically titrated in 1-2 min: 20-30 determinations can be performed per hour. For substances containing nitrogen in the per cent range the relative standard deviations for eight different substances were 0.1-1%.

  9. A Novel Rapid Method to Determine Cannabis Seed Germination Ability

    OpenAIRE

    吉澤, 政夫; 荒金, 眞佐子; 鈴木, 幸子; 北川, 重美; 中嶋, 順一; 森, 謙一郎; 荻野, 周三; Masao, Yoshizawa; Masako, Aragane; Yukiko, Suzuki; Shigemi, Kitagawa; Jun'ichi, Nakajima; Ken'ichiro, Mori; Shuzo, Ogino; 東京都健康安全研究センター薬用植物園

    2011-01-01

    A novel rapid method to determine cannabis seed germination ability was developed. Cannabis seeds were soaked in water for 15 minutes at 40℃, and germs were removed from the seeds by cracking the shell with tweezers. The germs were soaked in water for 15 minutes at 40℃, and the swollen skin was peeled off. The peeled germs were again soaked in water for 10 minutes at 40℃, and the angles between the seed leaves were measured. Cannabis seeds with angles of the seed leaves exceeding 30 degrees w...

  10. RAPID SEPARATION METHOD FOR EMERGENCY WATER AND URINE SAMPLES

    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, S.; Culligan, B.

    2008-08-27

    The Savannah River Site Environmental Bioassay Lab participated in the 2008 NRIP Emergency Response program administered by the National Institute for Standards and Technology (NIST) in May, 2008. A new rapid column separation method was used for analysis of actinides and {sup 90}Sr the NRIP 2008 emergency water and urine samples. Significant method improvements were applied to reduce analytical times. As a result, much faster analysis times were achieved, less than 3 hours for determination of {sup 90}Sr and 3-4 hours for actinides. This represents a 25%-33% improvement in analysis times from NRIP 2007 and a {approx}100% improvement compared to NRIP 2006 report times. Column flow rates were increased by a factor of two, with no significant adverse impact on the method performance. Larger sample aliquots, shorter count times, faster cerium fluoride microprecipitation and streamlined calcium phosphate precipitation were also employed. Based on initial feedback from NIST, the SRS Environmental Bioassay Lab had the most rapid analysis times for actinides and {sup 90}Sr analyses for NRIP 2008 emergency urine samples. High levels of potential matrix interferences may be present in emergency samples and rugged methods are essential. Extremely high levels of {sup 210}Po were found to have an adverse effect on the uranium results for the NRIP-08 urine samples, while uranium results for NRIP-08 water samples were not affected. This problem, which was not observed for NRIP-06 or NRIP-07 urine samples, was resolved by using an enhanced {sup 210}Po removal step, which will be described.

  11. A method for rapid similarity analysis of RNA secondary structures

    Directory of Open Access Journals (Sweden)

    Liu Na

    2006-11-01

    Full Text Available Abstract Background Owing to the rapid expansion of RNA structure databases in recent years, efficient methods for structure comparison are in demand for function prediction and evolutionary analysis. Usually, the similarity of RNA secondary structures is evaluated based on tree models and dynamic programming algorithms. We present here a new method for the similarity analysis of RNA secondary structures. Results Three sets of real data have been used as input for the example applications. Set I includes the structures from 5S rRNAs. Set II includes the secondary structures from RNase P and RNase MRP. Set III includes the structures from 16S rRNAs. Reasonable phylogenetic trees are derived for these three sets of data by using our method. Moreover, our program runs faster as compared to some existing ones. Conclusion The famous Lempel-Ziv algorithm can efficiently extract the information on repeated patterns encoded in RNA secondary structures and makes our method an alternative to analyze the similarity of RNA secondary structures. This method will also be useful to researchers who are interested in evolutionary analysis.

  12. A rapid and specific colorimetric method for free tryptophan quantification.

    Science.gov (United States)

    Wu, Yinan; Wang, Tianmin; Zhang, Chong; Xing, Xin-Hui

    2018-01-01

    Tryptophan is one of the eight essential amino acids and plays an important role in many biological processes. For its interaction with human health, environment and relevant commercial interest in biotechnology-based production, rapid and specific quantification method for this molecule accessible to common laboratories is badly needed. We herein reported a simple colorimetric method for free tryptophan quantification with 96-well-plate-level throughput. Our protocol firstly converted tryptophan to indole enzymatically by purified tryptophanases and then used reactivity of indole with hydroxylamine to form pink product with absorption peak at 530nm, enabling the quantification of tryptophan with simple spectrometry in just two hours. We presented that this method exhibited a linear detection range from 100μM to 600μM (R(2) = 0.9969) with no detection towards other naturally occurring tryptophan analogs or tryptophan residues in proteins. It was very robust in complicated biological samples, as demonstrated by quantifying the titer of 36 mutated tryptophan-producing strains with Pearson correlation coefficient of 0.93 in contrast to that measured by high performance liquid chromatography (HPLC). Our method should be potent for routine free tryptophan quantification in a high-throughput manner, facilitating studies in medicine, microbiology, food chemistry, metabolic engineering, etc. Copyright © 2017. Published by Elsevier B.V.

  13. Measurement of serum albumin by capillary zone electrophoresis, bromocresol green, bromocresol purple, and immunoassay methods

    Science.gov (United States)

    Duly, E B; Grimason, S; Grimason, P; Barnes, G; Trinick, T R

    2003-01-01

    Background/Aims: The introduction of capillary zone electrophoresis (CZE) to this laboratory has highlighted discrepancies in albumin measured on an Abbott Aeroset by bromocresol green (BCG) and that calculated by CZE on the basis of total protein measured by Biuret. Methods: This study examined differences in albumin estimation by CZE, Abbott Aeroset BCG, and Aeroset bromocresol purple (BCP), and compared these with albumin estimated by Beckman Array immunoassay. Results: Altman and Bland analysis of results showed a positive bias of BCG with CZE (4.51 g/litre; 95% limits of agreement, 3.77 to 5.26; n = 72) and BCP (3.85 g/litre; 95% limits of agreement, −1.42 to 9.12; n = 72). CZE and BCP agreed closely (0.67 g/litre; 95% limits of agreement, −4.39 to 3.06; n = 72). Analysis of 57 of those samples in which BCG and CZE differed ≥ 5 g/litre showed a positive bias of BCG with immunoassay (8.35 g/litre; 95% limits of agreement, 1.54 to 15.16; n =57), with good agreement between CZE and immunoassay (−0.44 g/litre; 95% limits of agreement, −2.82 to 1.94; n = 57). Conclusions: BCP is superior to BCG for the assay of albumin and has replaced BCG as the routine test for albumin in this laboratory. PMID:14514785

  14. Rapid identification of cytokinins by an immunological method

    Energy Technology Data Exchange (ETDEWEB)

    Morris, R.O.; Jameson, P.E.; Morris, J.W. (Univ. of Missouri-Columbia (USA)); Laloue, M. (Centre Nationale de la Recherche Scientifique, Gif-sur-Yvette (France))

    1991-04-01

    A method for rapid identification of bacterial cytokinins has been developed in which cultures are fed ({sup 3}H)adenine, the cytokinins (including, {sup 3}H-labeled cytokinins) are isolated by immunoaffinity chromatography, and analyzed by HPLC with on-line scintillation counting. Analysis of Agrobacterium tumefaciens strains showed that some produced primarily trans-zeatin, whereas others produced primarily trans-zeatin riboside. Pseudomonas syringae pv savastanoi produced mixtures of transzeatin, dihydrozeatin, 1{double prime}-methyl-trans-zeatin riboside, and other unknown cytokinin-like substances. Corynebacterium fascians, produced cis-zeatin, isopentenyladenine and isopentenyladenosine. The technique is designed for qualitative rather than quantitative studies and allows ready identification of bacterial cytokinins. It may also have utility in the study of plant cytokinins if adequate incorporation of label into cytokinin precursor pools can be achieved.

  15. A rapid protection switching method in carrier ethernet ring networks

    Science.gov (United States)

    Yuan, Liang; Ji, Meng

    2008-11-01

    Abstract: Ethernet is the most important Local Area Network (LAN) technology since more than 90% data traffic in access layer is carried on Ethernet. From 10M to 10G, the improving Ethernet technology can be not only used in LAN, but also a good choice for MAN even WAN. MAN are always constructed in ring topology because the ring network could provide resilient path protection by using less resource (fibre or cable) than other network topologies. In layer 2 data networks, spanning tree protocol (STP) is always used to protect transmit link and preventing the formation of logic loop in networks. However, STP cannot guarantee the efficiency of service convergence when link fault happened. In fact, convergent time of networks with STP is about several minutes. Though Rapid Spanning Tree Protocol (RSTP) and Multi-Spanning Tree Protocol (MSTP) improve the STP technology, they still need a couple of seconds to achieve convergence, and can not provide sub-50ms protection switching. This paper presents a novel rapid ring protection method (RRPM) for carrier Ethernet. Unlike other link-fault detection method, it adopts distributed algorithm to detect link fault rapidly (sub-50ms). When networks restore from link fault, it can revert to the original working state. RRPM can provide single ring protection and interconnected ring protection without the formation of super loop. In normal operation, the master node blocks the secondary port for all non-RRPM Ethernet frames belonging to the given RRPM Ring, thereby avoiding a loop in the ring. When link fault happens, the node on which the failure happens moves from the "ring normal" state to the "ring fault" state. It also sends "link down" frame immediately to other nodes and blocks broken port and flushes its forwarding database. Those who receive "link down" frame will flush forwarding database and master node should unblock its secondary port. When the failure restores, the whole ring will revert to the normal state. That is

  16. Method for Rapid Protein Identification in a Large Database

    Directory of Open Access Journals (Sweden)

    Wenli Zhang

    2013-01-01

    Full Text Available Protein identification is an integral part of proteomics research. The available tools to identify proteins in tandem mass spectrometry experiments are not optimized to face current challenges in terms of identification scale and speed owing to the exponential growth of the protein database and the accelerated generation of mass spectrometry data, as well as the demand for nonspecific digestion and post-modifications in complex-sample identification. As a result, a rapid method is required to mitigate such complexity and computation challenges. This paper thus aims to present an open method to prevent enzyme and modification specificity on a large database. This paper designed and developed a distributed program to facilitate application to computer resources. With this optimization, nearly linear speedup and real-time support are achieved on a large database with nonspecific digestion, thus enabling testing with two classical large protein databases in a 20-blade cluster. This work aids in the discovery of more significant biological results, such as modification sites, and enables the identification of more complex samples, such as metaproteomics samples.

  17. RAPID TEST METHOD FOR EVALUATION OF ANTIFREEZE ADDITIVE EFFICIENCY

    Directory of Open Access Journals (Sweden)

    S. V. Gushchin

    2015-01-01

    Full Text Available Usage of chemical additives while executing concrete works at negative temperatures is considered as a convenient and economical method. Range of the used antifreeze additives is rather wide. A great number of new additives are advertised but their characteristics have not been practically studied. Evaluation of the antifreeze additive efficiency is unfortunately rather long process and it does not provide comprehensive data on concrete structure formation processes. Due to this development of rapid and comprehensive methodology for construction companies is urgently required.Freezing processes of antifreeze additive aqueous solutions and hardening of cement paste with them have been investigated in the paper. The paper proposes a methodology for determination of freezing point for aqueous solutions of chemical additives of various applications. Identity of  freezing point for a chemical additive aqueous solution and cement paste with an equal concentration of the additive in the paste pore fluid has been determined while taking  calcium nitrate and sodium formate additives as an example. The paper demonstrates the possibility to evaluate efficiency of antifreeze additive action on the basis of kinetics in temperature changes of the cement paste with additives by its consecutive freezing and defrosting.  A methodology for operational evaluation in the field of chemical additive application for concreting items at negative temperatures has been offered in the paper.  The methodology does not require  deficient and expensive test-equipment. It can be applied at ordinary construction companies and it is comprehensible for personnel of low-qualification.  The paper shows the possibility to develop an original methodology for designing concrete structure which is based on operating efficiency determinations  for single and integrated antifreeze additives.

  18. A new rapid method for rockfall energies and distances estimation

    Science.gov (United States)

    Giacomini, Anna; Ferrari, Federica; Thoeni, Klaus; Lambert, Cedric

    2016-04-01

    and distances at the base to block and slope features. The validation of the proposed approach was conducted by comparing predictions to experimental data collected in the field and gathered from the scientific literature. The method can be used for both natural and constructed slopes and easily extended to more complicated and articulated slope geometries. The study shows its great potential for a quick qualitative hazard assessment providing indication about impact energy and horizontal distance of the first impact at the base of a rock cliff. Nevertheless, its application cannot substitute a more detailed quantitative analysis required for site-specific design of mitigation measures. Acknowledgements The authors gratefully acknowledge the financial support of the Australian Coal Association Research Program (ACARP). References Dorren, L.K.A. (2003) A review of rockfall mechanics and modelling approaches, Progress in Physical Geography 27(1), 69-87. Agliardi, F., Crosta, G.B., Frattini, P. (2009) Integrating rockfall risk assessment and countermeasure design by 3D modelling techniques. Natural Hazards and Earth System Sciences 9(4), 1059-1073. Ferrari, F., Thoeni, K., Giacomini, A., Lambert, C. (2016) A rapid approach to estimate the rockfall energies and distances at the base of rock cliffs. Georisk, DOI: 10.1080/17499518.2016.1139729.

  19. Rapid Methods for detection of Veterinary Drug residues in Meat

    Directory of Open Access Journals (Sweden)

    Chandan

    2010-10-01

    methodologies, the variety of residues to search per sample and the need to invest on powerful new instruments for identification and confirmatory purposes. Rapid and versatile screening methodologies make its control easier and reduce the number of non-compliant samples to be confirmed through tedious and costly confirmatory analytical methodologies. For instance, the multiresidue analysis can be performed better by using fast LC methods. Thus, the availability of new screening methodologies and the improvement of the existing ones will contribute to a better safety assurance of meat and other foods of animal origin. [Vet. World 2010; 3(5.000: 241-246

  20. Rapid method for identification of transgenic fish zygosity

    Directory of Open Access Journals (Sweden)

    . Alimuddin

    2007-07-01

    Full Text Available Identification of zygosity in transgenik fish is normally achieved by PCR analysis with genomic DNA template extracted from the tissue of progenies which are derived by mating the transgenic fish and wild-type counterpart.  This method needs relatively large amounts of fish material and is time- and labor-intensive. New approaches addressing this problem could be of great help for fish biotechnologists.  In this experiment, we applied a quantitative real-time PCR (qr-PCR method to analyze zygosity in a stable line of transgenic zebrafish (Danio rerio carrying masu salmon, Oncorhynchus masou D6-desaturase-like gene. The qr-PCR was performed using iQ SYBR Green Supermix in the iCycler iQ Real-time PCR Detection System (Bio-Rad Laboratories, USA.  Data were analyzed using the comparative cycle threshold method.  The results demonstrated a clear-cut identification of all transgenic fish (n=20 classified as a homozygous or heterozygous.  Mating of those fish with wild-type had revealed transgene transmission to the offspring following expected Mendelian laws. Thus, we found that the qTR-PCR to be effective for a rapid and precise determination of zygosity in transgenic fish. This technique could be useful in the establishment of breeding programs for mass transgenic fish production and in experiments in which zygosity effect could have a functional impact. Keywords: quantitative real-time PCR; zygosity; transgenic fish; mass production   ABSTRAK Identifikasi sigositas ikan transgenik biasanya dilakukan menggunakan analisa PCR dengan cetakan DNA genomik yang diekstraksi dari jaringan ikan hasil persilangan antara ikan transgenik dan ikan normal.   Metode ini memerlukan ikan dalam jumlah yang banyak, dan juga waktu serta tenaga.  Pendekatan baru untuk mengatasi masalah tersebut akan memberikan manfaat besar kepada peneliti bioteknologi perikanan.  Pada penelitian ini, kami menggunakan metode PCR real-time kuantitatif (krt-PCR untuk

  1. A rapid cloning method employing orthogonal end protection

    NARCIS (Netherlands)

    Jakobi, A.J.|info:eu-repo/dai/nl/311489621; Huizinga, E.G.|info:eu-repo/dai/nl/12314969X

    2012-01-01

    We describe a novel in vitro cloning strategy that combines standard tools in molecular biology with a basic protecting group concept to create a versatile framework for the rapid and seamless assembly of modular DNA building blocks into functional open reading frames. Analogous to chemical

  2. Rapid Radiochemical Method for Americium-241 in Building ...

    Science.gov (United States)

    Technical Fact Sheet Analysis Purpose: Qualitative analysis Technique: Alpha spectrometry Method Developed for: Americium-241 in building materials Method Selected for: SAM lists this method for qualitative analysis of americium-241 in concrete or brick building materials. Summary of subject analytical method which will be posted to the SAM website to allow access to the method.

  3. Rapid Radiochemical Method for Radium-226 in Building ...

    Science.gov (United States)

    Technical Fact Sheet Analysis Purpose: Qualitative analysis Technique: Alpha spectrometry Method Developed for: Radium-226 in building materials Method Selected for: SAM lists this method for qualitative analysis of radium-226 in concrete or brick building materials Summary of subject analytical method which will be posted to the SAM website to allow access to the method.

  4. Rapid method for plutonium-241 determination in soil samples

    OpenAIRE

    Piekarz, M.; Komosa, A.

    2014-01-01

    A simple and rapid procedure for the determination of plutonium isotopes in the environment is presented. The procedure combines alpha spectrometry, solvent extraction and liquid scintillation measurements to ensure that both alpha- and beta-emitting isotopes are determined. Of five tested extractants, bis-(2-ethylhexyl) phosphoric acid was found to be the best choice. The procedure was applied to soil samples contaminated with Chernobyl fallout.

  5. Wybrane Aspekty Zużywania Się Połączeń Wpustowych I Wielowypustowych W Procesie Eksploatacji

    Directory of Open Access Journals (Sweden)

    Gębura Andrzej

    2014-12-01

    Full Text Available W artykule przedstawiono wybrane wady połączeń wielowypustowych: mimośrodowości, przekoszenia, zwiększonych luzów. Przedstawiono znaczenie tych wad dla poprawności funkcjonowania całego zespołu napędowego. Zaprezentowano swoisty katalog tych wad w formie: wizualnego zużycia, pomiarów mechanicznych płaszczyzn przyporu, opisu matematycznego przebiegów dynamiki ruchu takiego połączenia, zobrazowań dla metod FAM-C i FDM-A. Zaznaczono skutki długotrwałej eksploatacji połączeń wielowypustowych o nadmiernym zużyciu dla bezpieczeństwa latania

  6. A rapid, small-scale sedimentation method to predict breadmaking quality of hard winter wheat

    Science.gov (United States)

    Breeders and processors are always looking for rapid and accurate methods to evaluate wheat quality. A rapid small-scale hybrid sedimentation method was developed for predicting breadmaking quality of breeders samples by combining the sodium dodecyl-sulfate (SDS) sedimentation method (AACC 56-70) an...

  7. A rapid method to estimate Westergren sedimentation rates

    Science.gov (United States)

    Alexy, Tamas; Pais, Eszter; Meiselman, Herbert J.

    2009-01-01

    The erythrocyte sedimentation rate (ESR) is a nonspecific but simple and inexpensive test that was introduced into medical practice in 1897. Although it is commonly utilized in the diagnosis and follow-up of various clinical conditions, ESR has several limitations including the required 60 min settling time for the test. Herein we introduce a novel use for a commercially available computerized tube viscometer that allows the accurate prediction of human Westergren ESR rates in as little as 4 min. Owing to an initial pressure gradient, blood moves between two vertical tubes through a horizontal small-bore tube and the top of the red blood cell (RBC) column in each vertical tube is monitored continuously with an accuracy of 0.083 mm. Using data from the final minute of a blood viscosity measurement, a sedimentation index (SI) was calculated and correlated with results from the conventional Westergren ESR test. To date, samples from 119 human subjects have been studied and our results indicate a strong correlation between SI and ESR values (R2=0.92). In addition, we found a close association between SI and RBC aggregation indices as determined by an automated RBC aggregometer (R2=0.71). Determining SI on human blood is rapid, requires no special training and has minimal biohazard risk, thus allowing physicians to rapidly screen for individuals with elevated ESR and to monitor therapeutic responses. PMID:19791973

  8. Comparison of rapid colorimetric method with conventional method in the isolation of mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Oberoi A

    2004-01-01

    Full Text Available The aim of the study was to evaluate two methods (colorimetric and conventional for isolation of Mycobacterium tuberculosis. A total of 500 clinical specimens were processed by modified Petroff′s method and then inoculated into MB/BacT-240 system bottles and on LJ medium slopes. The specimens included 242 sputum, 95 gastric aspirates, 47 pleural fluids, 45 CSF, 32 urine, 18 pus, 11 bronchoalveolar lavage, 3 tissue, 2 stool, 2 lymphnode specimens, 2 synovial fluid and 1 bronchial wash specimens. The isolation rate was 16.4% by the colorimetric method and 2.2% by the conventional method. The mean detection time was 16 days and 26 days respectively. Among 36 direct smear positive samples, 63.9%(23/36 and 30%(11/36 were positive by colorimetric and conventional methods respectively. Out of 464 direct smear negative samples 12.9%(60/464 and 0.6%(3/464 were positive by colorimetric and conventional methods respectively. Therefore, colorimetric method enables rapid detection leading to early diagnosis and drug susceptibility testing.

  9. Rapid and Reliable HPLC Method for the Determination of Vitamin ...

    African Journals Online (AJOL)

    Purpose: To develop and validate an accurate, sensitive and reproducible high performance liquid chromatographic (HPLC) method for the quantitation of vitamin C in pharmaceutical samples. Method: The drug and the standard were eluted from Superspher RP-18 (250 mm x 4.6 mm, 10ìm particle size) at 20 0C.

  10. Rapid, cost-effective liquid chromatograghic method for the ...

    African Journals Online (AJOL)

    The method is highly sensitive, with limit of detection of 1 ng/ml. The coefficient of variation for within-day run was less than 4% while that of day-to-day run was less than 6%. There were no interfering peaks from endogenous materials in the serum. The method was validated and used for pharmacokinetic studies ...

  11. A simple and rapid molecular method for Leptospira species identification

    NARCIS (Netherlands)

    Ahmed, Ahmed; Anthony, Richard M.; Hartskeerl, Rudy A.

    2010-01-01

    Serological and DNA-based classification systems only have little correlation. Currently serological and molecular methods for characterizing Leptospira are complex and costly restricting their world-wide distribution and use. Ligation mediated amplification combined with microarray analysis

  12. A method of rapid testing of radioactivity of different materials

    Directory of Open Access Journals (Sweden)

    Yu. Zabulonov

    2016-10-01

    Full Text Available A new method for the detection of low-level ionising radiation in solid, liquid or loose materials, which is based on the use of the Bayesian approach for the estimation of probabilistic parameters and a special statistical criterion, is offered in the present paper. We describe the algorithm and show the advantages of the method. The approach can be effective even in the case of extremely low signals whose intensity is much less than the background radiation.

  13. A rapid cloning method employing orthogonal end protection.

    Directory of Open Access Journals (Sweden)

    Arjen J Jakobi

    Full Text Available We describe a novel in vitro cloning strategy that combines standard tools in molecular biology with a basic protecting group concept to create a versatile framework for the rapid and seamless assembly of modular DNA building blocks into functional open reading frames. Analogous to chemical synthesis strategies, our assembly design yields idempotent composite synthons amenable to iterative and recursive split-and-pool reaction cycles. As an example, we illustrate the simplicity, versatility and efficiency of the approach by constructing an open reading frame composed of tandem arrays of a human fibronectin type III (FNIII domain and the von Willebrand Factor A2 domain (VWFA2, as well as chimeric (FNIII(n-VWFA2-(FNIII(n constructs. Although we primarily designed this strategy to accelerate assembly of repetitive constructs for single-molecule force spectroscopy, we anticipate that this approach is equally applicable to the reconstitution and modification of complex modular sequences including structural and functional analysis of multi-domain proteins, synthetic biology or the modular construction of episomal vectors.

  14. Rapid, specific and sensitive method for isoniazid determination in serum.

    Science.gov (United States)

    Sadeg, N; Pertat, N; Dutertre, H; Dumontet, M

    1996-01-12

    An original simple, specific and rapid high-performance liquid chromatographic assay for the determination of isoniazid (INH) in human serum is presented. The drug was extracted from the serum by protein precipitation with 30% (w/v) trichloroacetic acid, then the drug was reacted with the coupling reagent, trans-cinnamaldehyde, to form a derivative absorbing at 340 nm. A 20-microliters aliquot was injected into the chromatograph after neutralization with 1 M KOH solution. A liquid chromatograph equipped with a reversed-phase 30-microns C18 precolumn linked to a 4-microns C18 analytical column was used. The drug was eluted with a mixture of acetonitrile-water-triethylamine-acetic acid (400:600:2:1, v/v), pH value was 5 +/- 1. Flow-rate and wavelength were set at 1 ml/min and 340 nm, respectively. The extraction recoveries from human serum averaged 100% for INH at concentrations of 1, 2 and 4 mg/l. The coefficients of variation for three different concentrations for INH in serum in the within-day study varied between 1.2 and 3.5%, whereas those in the day-to-day study varied between 2.8 and 4.3%.

  15. Asymptotic and Numerical Methods for Rapidly Rotating Buoyant Flow

    Science.gov (United States)

    Grooms, Ian G.

    This thesis documents three investigations carried out in pursuance of a doctoral degree in applied mathematics at the University of Colorado (Boulder). The first investigation concerns the properties of rotating Rayleigh-Benard convection -- thermal convection in a rotating infinite plane layer between two constant-temperature boundaries. It is noted that in certain parameter regimes convective Taylor columns appear which dominate the dynamics, and a semi-analytical model of these is presented. Investigation of the columns and of various other properties of the flow is ongoing. The second investigation concerns the interactions between planetary-scale and mesoscale dynamics in the oceans. Using multiple-scale asymptotics the possible connections between planetary geostrophic and quasigeostrophic dynamics are investigated, and three different systems of coupled equations are derived. Possible use of these equations in conjunction with the method of superparameterization, and extension of the asymptotic methods to the interactions between mesoscale and submesoscale dynamics is ongoing. The third investigation concerns the linear stability properties of semi-implicit methods for the numerical integration of ordinary differential equations, focusing in particular on the linear stability of IMEX (Implicit-Explicit) methods and exponential integrators applied to systems of ordinary differential equations arising in the numerical solution of spatially discretized nonlinear partial differential equations containing both dispersive and dissipative linear terms. While these investigations may seem unrelated at first glance, some reflection shows that they are in fact closely linked. The investigation of rotating convection makes use of single-space, multiple-time-scale asymptotics to deal with dynamics strongly constrained by rotation. Although the context of thermal convection in an infinite layer seems somewhat removed from large-scale ocean dynamics, the asymptotic

  16. Rapid, cost-effective liquid chromatograghic method for the ...

    African Journals Online (AJOL)

    GRACE

    2006-07-03

    Jul 3, 2006 ... Akay C, Ozkan SA, Senturk Z, Cevheroglu S (2002). Simultaneous determination of metronidazole and miconazole in pharmaceutical dosage form by HPLC. Farmaco. Nov.57(11): 953-7. Galmier MJ, Frasey AM, Bastide M, Beyssac E, Petit J, Aiache JM,. Lartigue-Mattei (1998). Simple and sensitive method ...

  17. RESEARCH NOTE A Universal, rapid, and inexpensive method for ...

    Indian Academy of Sciences (India)

    Navya

    very satisfactory for many researchers to prepare reagents for “all in one” ready to use method to extract gDNA from very wide range sources of blood samples. To meet these criteria, a universal and versatile DNA extraction procedure should be developedwith a minimal chemicals and equipment.On the other hand, the ...

  18. Rapid prototyping methods for the manufacture of fuel cells

    Directory of Open Access Journals (Sweden)

    Dudek Piotr

    2016-01-01

    The potential for the application of this method for the manufacture of metallic bipolar plates (BPP for use in proton exchange membrane fuel cells (PEMFCs is presented and discussed. Special attention is paid to the fabrication of light elements for the construction of PEMFC stacks designed for mobile applications such as aviation technology and unmanned aerial vehicles (UAVs.

  19. [Methods for the rapid preparation of paraffin blocks].

    Science.gov (United States)

    Shmurun, R I

    1992-01-01

    Two accelerated chloroform-paraffin processings of materials with the use of ultrasound (US) and microwave (MW) irradiation in the stove "Electronica" as well as a combined method with US- and MW-irradiation are proposed to shorten drastically the duration of the prehistologic processing.

  20. Rapid multi-residue method for the determination of pesticide ...

    African Journals Online (AJOL)

    Exposure to pesticides can represent a potential risk to humans. Agricultural workers are at risk of chronic toxicity. Hence, the evaluation of pesticide residues in their blood gives an indication about the extent of exposure and help in assessing adverse health effects. The aim of our study was to develop analytical method for ...

  1. Time evolution of the wave equation using rapid expansion method

    KAUST Repository

    Pestana, Reynam C.

    2010-07-01

    Forward modeling of seismic data and reverse time migration are based on the time evolution of wavefields. For the case of spatially varying velocity, we have worked on two approaches to evaluate the time evolution of seismic wavefields. An exact solution for the constant-velocity acoustic wave equation can be used to simulate the pressure response at any time. For a spatially varying velocity, a one-step method can be developed where no intermediate time responses are required. Using this approach, we have solved for the pressure response at intermediate times and have developed a recursive solution. The solution has a very high degree of accuracy and can be reduced to various finite-difference time-derivative methods, depending on the approximations used. Although the two approaches are closely related, each has advantages, depending on the problem being solved. © 2010 Society of Exploration Geophysicists.

  2. A taxonomy of rapid reviews links report types and methods to specific decision-making contexts.

    Science.gov (United States)

    Hartling, Lisa; Guise, Jeanne-Marie; Kato, Elisabeth; Anderson, Johanna; Belinson, Suzanne; Berliner, Elise; Dryden, Donna M; Featherstone, Robin; Mitchell, Matthew D; Motu'apuaka, Makalapua; Noorani, Hussein; Paynter, Robin; Robinson, Karen A; Schoelles, Karen; Umscheid, Craig A; Whitlock, Evelyn

    2015-12-01

    Describe characteristics of rapid reviews and examine the impact of methodological variations on their reliability and validity. We conducted a literature review and interviews with organizations that produce rapid reviews or related products to identify methods, guidance, empiric evidence, and current practices. We identified 36 rapid products from 20 organizations (production time, 5 minutes to 8 months). Methods differed from systematic reviews at all stages. As time frames increased, methods became more rigorous; however, restrictions on database searching, inclusion criteria, data extracted, and independent dual review remained. We categorized rapid products based on extent of synthesis. "Inventories" list what evidence is available. "Rapid responses" present best available evidence with no formal synthesis. "Rapid reviews" synthesize the quality of and findings from the evidence. "Automated approaches" generate meta-analyses in response to user-defined queries. Rapid products rely on a close relationship with end users and support specific decisions in an identified time frame. Limited empiric evidence exists comparing rapid and systematic reviews. Rapid products have tremendous methodological variation; categorization based on time frame or type of synthesis reveals patterns. The similarity across rapid products lies in the close relationship with the end user to meet time-sensitive decision-making needs. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Systems and methods for rapid processing and storage of data

    Science.gov (United States)

    Stalzer, Mark A.

    2017-01-24

    Systems and methods of building massively parallel computing systems using low power computing complexes in accordance with embodiments of the invention are disclosed. A massively parallel computing system in accordance with one embodiment of the invention includes at least one Solid State Blade configured to communicate via a high performance network fabric. In addition, each Solid State Blade includes a processor configured to communicate with a plurality of low power computing complexes interconnected by a router, and each low power computing complex includes at least one general processing core, an accelerator, an I/O interface, and cache memory and is configured to communicate with non-volatile solid state memory.

  4. Rapid new methods for paint collection and lead extraction.

    Science.gov (United States)

    Gutknecht, William F; Harper, Sharon L; Winstead, Wayne; Sorrell, Kristen; Binstock, David A; Salmons, Cynthia A; Haas, Curtis; McCombs, Michelle; Studabaker, William; Wall, Constance V; Moore, Curtis

    2009-01-01

    Chronic exposure of children to lead can result in permanent physiological impairment. In adults, it can cause irritability, poor muscle coordination, and nerve damage to the sense organs and nerves controlling the body. Surfaces coated with lead-containing paints are potential sources of exposure to lead. In April 2008, the U.S. Environmental Protection Agency (EPA) finalized new requirements that would reduce exposure to lead hazards created by renovation, repair, and painting activities, which disturb lead-based paint. On-site, inexpensive identification of lead-based paint is required. Two steps have been taken to meet this challenge. First, this paper presents a new, highly efficient method for paint collection that is based on the use of a modified wood drill bit. Second, this paper presents a novel, one-step approach for quantitatively grinding and extracting lead from paint samples for subsequent lead determination. This latter method is based on the use of a high-revolutions per minute rotor with stator to break up the paint into approximately 50 micron-size particles. Nitric acid (25%, v/v) is used to extract the lead in 95% for real-world paints, National Institute of Standards and Technology's standard reference materials, and audit samples from the American Industrial Hygiene Association's Environmental Lead Proficiency Analytical Testing Program. This quantitative extraction procedure, when paired with quantitative paint sample collection and lead determination, may enable the development of a lead paint test kit that will meet the specifications of the final EPA rule.

  5. Susceptibility testing of filamentous fungi to amphotericin B by a rapid radiometric method

    Energy Technology Data Exchange (ETDEWEB)

    Merz, W.G.; Fay, D.; Thumar, B.; Dixon, D.

    1984-01-01

    A rapid, radiometric method was developed to determine the susceptibility of filamentous fungi to amphotericin B. The rapid, radiometric method depended on measurement of the inhibition of /sup 24/CO/sub 2/ production in the presence of amphotericin B. Thirty isolates of filamentous fungi were tested by the rapid, radiometric method and a reference agar dilution method. There was 93% agreement between the two methods when an 80% or greater decrease in CO/sub 2/ production was used to calculate the minimal inhibitory concentration with the rapid, radiometric method. Minimal inhibitory concentrations, based on 80% decrease of CO/sub 2/ production, were achieved within 24 h of incubation with all of the fungi tested.

  6. Microwave as a rapid cooking method for beef tenderness evaluation.

    Science.gov (United States)

    Silva, Douglas R G; Fernandez, Ludimila C; Torres Filho, Robledo A; Fontes, Paulo R; Ramos, Alcinéia L S; Ramos, Eduardo M

    2017-11-20

    Semitendinosus (ST) muscle steaks were grouped according to three locations (proximal, middle, and distal end), grilled to endpoint temperature of 71C or cooked for 20, 30, 40, 50, or 60 s in a microwave oven (Mw). The location did not affect (p > .05) the cooking loss (CL) or shear force (SF) values. The CL increased (p  .05) from the grill samples. None of the microwaves' SF values were different (p > .05) from the grill values, with treatments Mw30 to Mw50 showing moderate repeatability (R = 0.51-0.60) and Mw30 and Mw60 showing higher correlations (r > .71) with grill values. Cooking beef strips with a microwave is a potential method for tenderness evaluation, but requires additional study to evaluate and optimize this application in different muscles and for comparison to sensorial data. The work was intended to evaluate the possibility of using a microwave oven for cooking meat to be used in objective measurement protocols for meat tenderness and to optimize the conditions for this purpose. The use of a standardized microwave procedure allows a dramatic reduction in analysis time and may reduce error variance due to nonuniform cooking procedures. © 2017 Wiley Periodicals, Inc.

  7. A method for rapid abundance estimation of semiplanktonic meiofauna

    Science.gov (United States)

    Armonies, Werner

    2000-12-01

    Many meiofaunal copepods and plathelminths enter the tidal waters at night thus exhibiting a life-style intermediate between benthic and planktonic. At the same time, ostracods may leave their interstitial dwelling and move across the sediment surface. In laboratory experiments, the percentage of plathelminth populations emerging from the sediment varied with the species, temperature, light conditions, and the dimensions of the sediment cores studied, but not with tidal level, season, ambient density of conspecifics, or the sediment composition. Therefore, the swimming activity may be utilised for extraction of semiplanktonic meiofauna provided that the extraction procedure is standardised with respect to temperature, light and core size. For free-living plathelminths from the Wadden Sea intertidal a robust standard procedure is as follows: sediment cores 1.6 cm in diameter (2 cm2 surface area) and 3 cm deep are fitted into cylindrical containers and submerged into aquaria containing filtered seawater (ambient salinity, room temperature, darkness) for 24 h. The sediment containers are then removed and the aquarian water filtered through appropriate meshes; the residue contains the emergent faunal component. For plathelminths, this procedure reduces sorting time by some 90% compared with the standard shaking-decantation method and thus makes it possible to process a high number of samples in a short time. Similar procedures may be developed for copepods and epibenthic ostracods.

  8. Rapid methods for the detection of foodborne bacterial pathogens: principles, applications, advantages and limitations

    Science.gov (United States)

    Law, Jodi Woan-Fei; Ab Mutalib, Nurul-Syakima; Chan, Kok-Gan; Lee, Learn-Han

    2015-01-01

    The incidence of foodborne diseases has increased over the years and resulted in major public health problem globally. Foodborne pathogens can be found in various foods and it is important to detect foodborne pathogens to provide safe food supply and to prevent foodborne diseases. The conventional methods used to detect foodborne pathogen are time consuming and laborious. Hence, a variety of methods have been developed for rapid detection of foodborne pathogens as it is required in many food analyses. Rapid detection methods can be categorized into nucleic acid-based, biosensor-based and immunological-based methods. This review emphasizes on the principles and application of recent rapid methods for the detection of foodborne bacterial pathogens. Detection methods included are simple polymerase chain reaction (PCR), multiplex PCR, real-time PCR, nucleic acid sequence-based amplification (NASBA), loop-mediated isothermal amplification (LAMP) and oligonucleotide DNA microarray which classified as nucleic acid-based methods; optical, electrochemical and mass-based biosensors which classified as biosensor-based methods; enzyme-linked immunosorbent assay (ELISA) and lateral flow immunoassay which classified as immunological-based methods. In general, rapid detection methods are generally time-efficient, sensitive, specific and labor-saving. The developments of rapid detection methods are vital in prevention and treatment of foodborne diseases. PMID:25628612

  9. Rapid Methods for the Detection of Foodborne Bacterial Pathogens: Principles, Applications, Advantages and Limitations

    Directory of Open Access Journals (Sweden)

    Law eJodi Woan-Fei

    2015-01-01

    Full Text Available The incidence of foodborne diseases has increased over the years and resulted in major public health problem globally. Foodborne pathogens can be found in various foods and it is important to detect foodborne pathogens to provide safe food supply and to prevent foodborne diseases. The conventional methods used to detect foodborne pathogen are time consuming and laborious. Hence, a variety of methods have been developed for rapid detection of foodborne pathogens as it is required in many food analyses. Rapid detection methods can be categorized into nucleic acid-based, biosensor-based and immunological-based methods. This review emphasizes on the principles and application of recent rapid methods for the detection of foodborne bacterial pathogens. Detection methods included are simple polymerase chain reaction (PCR, multiplex PCR, real-time PCR, nucleic acid sequence-based amplification (NASBA, loop-mediated isothermal amplification (LAMP and oligonucleotide DNA microarray which classified as nucleic acid-based methods; optical, electrochemical and mass-based biosensors which classified as biosensor-based methods; enzyme-linked immunosorbent assay (ELISA and lateral flow immunoassay which classified as immunological-based methods. In general, rapid detection methods are generally time-efficient, sensitive, specific and labor-saving. The developments of rapid detection methods are vital in prevention and treatment of foodborne diseases.

  10. Methods and Magnitudes of Rapid Weight Loss in Judo Athletes Over Pre-Competition Periods

    Directory of Open Access Journals (Sweden)

    Kons Rafael Lima

    2017-06-01

    Full Text Available Purpose. The study aimed to analyse the methods and magnitudes of rapid weight loss (RWL in judo team members in distinct periods before the biggest state competition in Southern Brazil.

  11. High efficiency joint CZE determination of sugars and acids in vegetables and fruits.

    Science.gov (United States)

    Cebolla-Cornejo, Jaime; Valcárcel, Mercedes; Herrero-Martínez, José Manuel; Roselló, Salvador; Nuez, Fernando

    2012-08-01

    In this work, an improved CE method for the medium-throughput determination of main organic acids (oxalate, malate, citrate), the amino acid glutamate and the sugars fructose, glucose and sucrose in several food matrices is described. These compounds have been identified as key components in the taste intensity of fruit and vegetable crops. Using a running buffer with 20 mM 2,6-pyridine dicarboxylic acid pH 12.1 and 0.1% hexadimethrine bromide, replacing it every 5 h to avoid pH decrease, and optimizing capillary conditioning between runs with 58 mM SDS during 2 min at 20 psi, it is possible to effectively quantify these compounds while increasing medium throughput repeatability. This procedure resolves problems such as increases in migration time and reduction of resolution between problematic peaks (malate/citrate and fructose/glucose) detected in a previous method. The new procedure even considerably reduced time analysis down to 12 min. Under optimal conditions, a large number of injections (200) could be administered without any disturbances in the same capillary. The reliability of the proposed method was further investigated with several food matrix samples, including tomato, pepper, muskmelon, winter squash, and orange. This method is recommended for routine analysis of large number of samples typical of production quality systems or plant breeding programs. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Rapid identification of salmonella serotypes with stereo and hyperspectral microscope imaging Methods

    Science.gov (United States)

    The hyperspectral microscope imaging (HMI) method can reduce detection time within 8 hours including incubation process. The early and rapid detection with this method in conjunction with the high throughput capabilities makes HMI method a prime candidate for implementation for the food industry. Th...

  13. Assessment of three rapid methods for the detection of methicillin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Soares, Maria João; Soares, Carlos; Mendes, Ana Constança; Guimarães, Maria Luís; Cabeda, José Manuel; Amorim, José Manuel

    2004-01-01

    We evaluated three rapid methods to detect methicillin-resistant Staphylococcus aureus (MRSA) and compared them with PCR amplification of mecA. A total of 103 S. aureus strains were studied by MRSA-Screen, BBL Crystal, Velogene Genomic and mecA PCR. All the methods detected the 61 MRSA strains having the mecA gene, showing 100% sensitivity and specificity. Despite the correlation between all the rapid methods and PCR, the ease of use and shorter turnaround time of MRSA-Screen were important factors leading to the selection of this method as the routine screening technique for MRSA.

  14. The scope of application of incremental rapid prototyping methods in foundry engineering

    Directory of Open Access Journals (Sweden)

    M. Stankiewicz

    2010-01-01

    Full Text Available The article presents the scope of application of selected incremental Rapid Prototyping methods in the process of manufacturing casting models, casting moulds and casts. The Rapid Prototyping methods (SL, SLA, FDM, 3DP, JS are predominantly used for the production of models and model sets for casting moulds. The Rapid Tooling methods, such as: ZCast-3DP, ProMetalRCT and VoxelJet, enable the fabrication of casting moulds in the incremental process. The application of the RP methods in cast production makes it possible to speed up the prototype preparation process. This is particularly vital to elements of complex shapes. The time required for the manufacture of the model, the mould and the cast proper may vary from a few to several dozen hours.

  15. Evaluation of two methods of rapid blood-glucose monitoring by unskilled personnel during surgery

    DEFF Research Database (Denmark)

    Madsbad, S; Adelhøj, B; Bigler, Dennis Richard

    1984-01-01

    The accuracy of two rapid methods of blood-glucose monitoring without (Haemo-glucotest 1-44) and with a reflectance meter (Hypocount B) was compared using a laboratory method. The assessment was carried out by personnel with no previous experience in measuring blood glucose. Eighty-five percent o...

  16. A rapid and convenient method for preparing salt-free (. gamma. -/sup 32/P)ATP

    Energy Technology Data Exchange (ETDEWEB)

    Palmer, J.L.; Avruch, J.

    1981-09-15

    (..gamma..-/sup 32/P)ATP is prepared by an existing enzymatic method that yields approximately 95% incorporation of /sup 32/P into ATP. A rapid and convenient method for purifying the (..gamma..-/sup 32/P)ATP which results in a product free of both salt and buffer is reported.

  17. High performance liquid chromatography method for rapid and accurate determination of homocysteine in plasma and serum

    DEFF Research Database (Denmark)

    Vester, Birte; Rasmussen, K

    1991-01-01

    Determination of homocysteine in plasma or serum for evaluation of cobalamin and folate deficiency is becoming an important diagnostic procedure. Accurate, rapid and low cost methods for measuring homocysteine are therefore required. We have improved an HPLC method and made it suitable for clinical...

  18. A rapid method to determine sterol, erythrodiol, and uvaol concentrations in olive oil.

    Science.gov (United States)

    Mathison, Brian; Holstege, Dirk

    2013-05-15

    A rapid, accurate, and efficient method for determining the sterol, uvaol, and erythrodiol concentrations was developed to meet International Olive Council (IOC) certification criteria for extra virgin olive oil (EVOO). The unsaponifiable fraction of the sample (0.2 g) was separated with a diatomaceous earth column, and the sterol and triterpenic dialcohols were isolated with a novel base-activated silica solid-phase extraction (SPE) cartridge cleanup protocol. The improved method and the IOC method provided identical pass/fail results (n = 34) for each of the six sterol and erythrodiol/uvaol IOC criteria used to assess olive oil. This method was validated, and recoveries of stigmasterol (88%) and β-sitosterol (84%) were greater than previously published values obtained using the IOC method. This method requires approximately one-third the time required to complete the IOC method and has great utility for the rapid screening of EVOO to detect adulteration, false labeling, and an inferior product.

  19. Commercially Available Rapid Methods for Detection of Selected Food-borne Pathogens.

    Science.gov (United States)

    Valderrama, Wladir B; Dudley, Edward G; Doores, Stephanie; Cutter, Catherine N

    2016-07-03

    Generally, the enumeration and isolation of food-borne pathogens is performed using culture-dependent methods. These methods are sensitive, inexpensive, and provide both qualitative and quantitative assessment of the microorganisms present in a sample, but these are time-consuming. For this reason, researchers are developing new techniques that allow detection of food pathogens in shorter period of time. This review identifies commercially available methods for rapid detection and quantification of Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, and Shiga toxin-producing Escherichia coli in food samples. Three categories are discussed: immunologically based methods, nucleic acid-based assays, and biosensors. This review describes the basic mechanism and capabilities of each method, discusses the difficulties of choosing the most convenient method, and provides an overview of the future challenges for the technology for rapid detection of microorganisms.

  20. Rapid high temperature field test method for evaluation of geothermal calcite scale inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Asperger, R.G.

    1982-08-01

    A test method is described which allows the rapid field testing of calcite scale inhibitors in high- temperature geothermal brines. Five commercial formulations, chosen on the basis of laboratory screening tests, were tested in brines with low total dissolved solids at ca 500 F. Four were found to be effective; of these, 2 were found to be capable of removing recently deposited scale. One chemical was tested in the full-flow brine line for 6 wks. It was shown to stop a severe surface scaling problem at the well's control valve, thus proving the viability of the rapid test method. (12 refs.)

  1. Clinical application of a rapid microbiological test based on capillary zone electrophoresis to assess local skin infection

    Directory of Open Access Journals (Sweden)

    Szeliga Jacek

    2011-10-01

    Full Text Available Abstract Background The basic clinical problem associated with infection treatment is the fact that classic, commonly and routinely used isolation and identification methods are based on long-term processes of a phenotypic analysis of microorganisms. Consequently sometimes, especially in small centres, rapid implementation of antibacterial treatment becomes delayed. The work presents the initial results of rapid microbiological identification based on an original method of capillary zone electrophoresis (CZE. The study involved the analysis of 78 biological samples from post-operative wounds and trophic ulcers. Results The attempt was made to identify individual bacterial species based on characteristic features of electropherograms achieved. Finally, G(+ cocci type bacteria and different G(- rods were identified with sensitivity of 88.1% and specificity of 100%. Conclusions Based on the clinical trials using an electrophoretic technique in the field of microbiological diagnostics of infected exudate from a post-operative wound it can be concluded that it is a rapid and relatively sensitive method for initial identification of infectious pathogens.

  2. A rapid method for soil cement design : Louisiana slope value method.

    Science.gov (United States)

    1964-03-01

    The current procedure used by the Louisiana Department of Highways for laboratory design of cement stabilized soil base and subbase courses is taken from standard AASHO test methods, patterned after Portland Cement Association criteria. These methods...

  3. Furuncular myiasis: a simple and rapid method for extraction of intact Dermatobia hominis larvae.

    Science.gov (United States)

    Boggild, Andrea K; Keystone, Jay S; Kain, Kevin C

    2002-08-01

    We report a case of furuncular myiasis complicated by Staphylococcus aureus infection and beta-hemolytic streptococcal cellulitis. The Dermatobia hominis larva that caused this lesion could not be extracted using standard methods, including suffocation and application of lateral pressure, and surgery was contraindicated because of cellulitis. The botfly maggot was completely and rapidly extracted with an inexpensive, disposable, commercial venom extractor.

  4. Collision-induced fragmentation accurate mass spectrometric analysis methods to rapidly characterize phytochemicals in plant extracts

    Science.gov (United States)

    The rapid advances in analytical chromatography equipment have made the reliable and reproducible measurement of a wide range of plant chemical components possible. Full chemical characterization of a given plant material is possible with the new mass spectrometers currently available. New methods a...

  5. Collaborative validation of a rapid method for efficient virus concentration in bottled water

    DEFF Research Database (Denmark)

    Schultz, Anna Charlotte; Perelle, Sylvie; Di Pasquale, Simona

    2011-01-01

    Enteric viruses, including norovirus (NoV) and hepatitis A virus (HAV), have emerged as a major cause of waterborne outbreaks worldwide. Due to their low infectious doses and low concentrations in water samples, an efficient and rapid virus concentration method is required for routine control. Th...

  6. A Rapid Method for Measuring Strontium-90 Activity in Crops in China

    Directory of Open Access Journals (Sweden)

    Pan Lingjing Pan

    2017-01-01

    Full Text Available A rapid method for measuring Sr-90 activity in crop ashes is presented. Liquid scintillation counting, combined with ion exchange columns 4‘, 4“(5“-di-t-butylcyclohexane-18-crown-6, is used to determine the activity of Sr-90 in crops. The yields of chemical procedure are quantified using gravimetric analysis. The conventional method that uses ion-exchange resin with HDEHP could not completely remove all the bismuth when comparatively large lead and bismuth exist in the samples. This is overcome by the rapid method. The chemical yield of this method is about 60% and the MDA for Sr-90 is found to be 2:32 Bq/kg. The whole procedure together with using spectrum analysis to determine the activity only takes about one day, which is really a large improvement compared with the conventional method. A modified conventional method is also described here to verify the value of the rapid one. These two methods can meet di_erent needs of daily monitoring and emergency situation.

  7. A Rapid Method for Measuring Strontium-90 Activity in Crops in China

    Science.gov (United States)

    Pan, Lingjing Pan; Yu, Guobing; Wen, Deyun; Chen, Zhi; Sheng, Liusi; Liu, Chung-King; Xu, X. George

    2017-09-01

    A rapid method for measuring Sr-90 activity in crop ashes is presented. Liquid scintillation counting, combined with ion exchange columns 4`, 4"(5")-di-t-butylcyclohexane-18-crown-6, is used to determine the activity of Sr-90 in crops. The yields of chemical procedure are quantified using gravimetric analysis. The conventional method that uses ion-exchange resin with HDEHP could not completely remove all the bismuth when comparatively large lead and bismuth exist in the samples. This is overcome by the rapid method. The chemical yield of this method is about 60% and the MDA for Sr-90 is found to be 2:32 Bq/kg. The whole procedure together with using spectrum analysis to determine the activity only takes about one day, which is really a large improvement compared with the conventional method. A modified conventional method is also described here to verify the value of the rapid one. These two methods can meet di_erent needs of daily monitoring and emergency situation.

  8. Considerations for Task Analysis Methods and Rapid E-Learning Development Techniques

    Directory of Open Access Journals (Sweden)

    Dr. Ismail Ipek

    2014-02-01

    Full Text Available The purpose of this paper is to provide basic dimensions for rapid training development in e-learning courses in education and business. Principally, it starts with defining task analysis and how to select tasks for analysis and task analysis methods for instructional design. To do this, first, learning and instructional technologies as visions of the future were discussed. Second, the importance of task analysis methods in rapid e-learning was considered, with learning technologies as asynchronous and synchronous e-learning development. Finally, rapid instructional design concepts and e-learning design strategies were defined and clarified with examples, that is, all steps for effective task analysis and rapid training development techniques based on learning and instructional design approaches were discussed, such as m-learning and other delivery systems. As a result, the concept of task analysis, rapid e-learning development strategies and the essentials of online course design were discussed, alongside learner interface design features for learners and designers.

  9. An economical and combined method for rapid and efficient isolation of fungal DNA.

    Science.gov (United States)

    Lech, T; Syguła-Cholewinska, J; Szostak-Kot, J

    2014-12-18

    DNA isolation is a crucial step of conducting genetic studies in any organism. However, this process is quite difficult when studying fungi because of the need to damage the fungal cell walls of specific structures. In this study, we developed a method for the rapid and efficient isolation of fungal DNA based on simultaneous mechanical and enzymatic cell wall degradation. There are several typical modifications of the standard phenol-chloroform DNA extraction method. This method can be modified to degrade the fungal cell wall. The first step of the presented DNA extraction included manual homogenization in modified lysis buffer. Next, enzymatic digestion using 2 enzymes was conducted, including lyticase and proteinase K. To carefully select the most favorable conditions, we developed an economical, rapid, and reliable method for fungal DNA extraction that ensures both high efficiency and proper purity, which are essential for further analyses.

  10. Rapid method for determination of carbonyl groups in lignin compounds by headspace gas chromatography.

    Science.gov (United States)

    Li, Jing; Hu, Hui-Chao; Chai, Xin-Sheng

    2015-07-24

    The paper reports on a novel method for rapid determination of carbonyl in lignins by headspace gas chromatography (HS-GC). The method involves the quantitative carbonyl reduction for aldehydes in 2min at room temperature or for acetones in 30min at 80°C by sodium borohydride solution in a closed headspace sample vial. After the reaction, the solution was acidified by injecting sulfuric acid solution and the hydrogen released to the headspace was determined by GC using thermal-conductivity detector. The results showed that with the addition of SiO2 powder, the reduction reaction of carbonyl groups can be greatly facilitated. The method has a good measurement precision (RSD<7.74%) and accuracy (relative error <10% compared with a reference method) in the carbonyl quantification. It is suitable to be used for rapid determination of carbonyl content in lignin and related materials. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Apparatus and method for rapid separation and detection of hydrocarbon fractions in a fluid stream

    Science.gov (United States)

    Sluder, Charles S.; Storey, John M.; Lewis, Sr., Samuel A.

    2013-01-22

    An apparatus and method for rapid fractionation of hydrocarbon phases in a sample fluid stream are disclosed. Examples of the disclosed apparatus and method include an assembly of elements in fluid communication with one another including one or more valves and at least one sorbent chamber for removing certain classifications of hydrocarbons and detecting the remaining fractions using a detector. The respective ratios of hydrocarbons are determined by comparison with a non separated fluid stream.

  12. A convenient and rapid method for genetic transformation of E. coli with plasmids.

    Science.gov (United States)

    Chen, X; Guo, P; Xie, Z; Shen, P

    2001-12-01

    A convenient and rapid method for the genetic transformation of Escherichia coli with plasmids is proposed. By mixing the recipient cells and plasmid DNA and spreading them directly on selective medium plates containing Ca2+, the so-called 'plate transformation' could achieve almost the same transformation efficiency as the classical transformation method with calcium. The whole protocol takes only about 2 min, its simplicity compared favorably, not only to the usual protocol, but also to all other documented modifications.

  13. Rapid qualitative research methods during complex health emergencies: A systematic review of the literature.

    Science.gov (United States)

    Johnson, Ginger A; Vindrola-Padros, Cecilia

    2017-09-01

    The 2013-2016 Ebola outbreak in West Africa highlighted both the successes and limitations of social science contributions to emergency response operations. An important limitation was the rapid and effective communication of study findings. A systematic review was carried out to explore how rapid qualitative methods have been used during global heath emergencies to understand which methods are commonly used, how they are applied, and the difficulties faced by social science researchers in the field. We also asses their value and benefit for health emergencies. The review findings are used to propose recommendations for qualitative research in this context. Peer-reviewed articles and grey literature were identified through six online databases. An initial search was carried out in July 2016 and updated in February 2017. The PRISMA checklist was used to guide the reporting of methods and findings. The articles were assessed for quality using the MMAT and AACODS checklist. From an initial search yielding 1444 articles, 22 articles met the criteria for inclusion. Thirteen of the articles were qualitative studies and nine used a mixed-methods design. The purpose of the rapid studies included: the identification of causes of the outbreak, and assessment of infrastructure, control strategies, health needs and health facility use. The studies varied in duration (from 4 days to 1 month). The main limitations identified by the authors were: the low quality of the collected data, small sample sizes, and little time for cross-checking facts with other data sources to reduce bias. Rapid qualitative methods were seen as beneficial in highlighting context-specific issues that need to be addressed locally, population-level behaviors influencing health service use, and organizational challenges in response planning and implementation. Recommendations for carrying out rapid qualitative research in this context included the early designation of community leaders as a point of

  14. Seed oil polyphenols: rapid and sensitive extraction method and high resolution-mass spectrometry identification.

    Science.gov (United States)

    Koubaa, Mohamed; Mhemdi, Houcine; Vorobiev, Eugène

    2015-05-01

    Phenolic content is a primary parameter for vegetables oil quality evaluation, and directly involved in the prevention of oxidation and oil preservation. Several methods have been reported in the literature for polyphenols extraction from seed oil but the approaches commonly used remain manually handled. In this work, we propose a rapid and sensitive method for seed oil polyphenols extraction and identification. For this purpose, polyphenols were extracted from Opuntia stricta Haw seed oil, using high frequency agitation, separated, and then identified using a liquid chromatography-high resolution mass spectrometry method. Our results showed good sensitivity and reproducibility of the developed methods. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. RAPID AND EFFICIENT METHOD FOR ENVIRONMENTAL DNA EXTRACTION AND PURIFICATION FROM SOIL

    Directory of Open Access Journals (Sweden)

    J. Hamedi

    2016-06-01

    Full Text Available Large proportion of microbial population in the world is unculturable. Extraction of total DNA from soil is usually a crucial step considering to the difficulties of study the uncultivable microorganisms. Humic acid is considered as the main inhibitory agent in the environmental DNA studies. Here, we introduced a rapid and efficient method for DNA extraction and purification from soil. Yield of DNA extraction by the presented method was 130 ng/µl. Three conventional methods of DNA extraction including liquid nitrogen incursion, bead beating and sonication were performed as control methods. Yield of DNA extraction by these methods were 110, 90 and 50 ng/µl, respectively. A rapid and efficient one step DNA purification method was introduced instead of hazardous conventional phenol-chloroform methods. Humic acid removal percentage by the introduced method was 95.8 % that is comparable with 97 % gained by the conventional gel extraction method and yield of DNA after purification was 84 % and 73 %, respectively. This study could be useful in molecular ecology and metagenomics study as a fast and reliable method.

  16. The use of rapid review methods in health technology assessments: 3 case studies.

    Science.gov (United States)

    Kaltenthaler, Eva; Cooper, Katy; Pandor, Abdullah; Martyn-St James, Marrissa; Chatters, Robin; Wong, Ruth

    2016-08-26

    Rapid reviews are of increasing importance within health technology assessment due to time and resource constraints. There are many rapid review methods available although there is little guidance as to the most suitable methods. We present three case studies employing differing methods to suit the evidence base for each review and outline some issues to consider when selecting an appropriate method. Three recently completed systematic review short reports produced for the UK National Institute for Health Research were examined. Different approaches to rapid review methods were used in the three reports which were undertaken to inform the commissioning of services within the NHS and to inform future trial design. We describe the methods used, the reasoning behind the choice of methods and explore the strengths and weaknesses of each method. Rapid review methods were chosen to meet the needs of the review and each review had distinctly different challenges such as heterogeneity in terms of populations, interventions, comparators and outcome measures (PICO) and/or large numbers of relevant trials. All reviews included at least 10 randomised controlled trials (RCTs), each with numerous included outcomes. For the first case study (sexual health interventions), very diverse studies in terms of PICO were included. P-values and summary information only were presented due to substantial heterogeneity between studies and outcomes measured. For the second case study (premature ejaculation treatments), there were over 100 RCTs but also several existing systematic reviews. Data for meta-analyses were extracted directly from existing systematic reviews with new RCT data added where available. For the final case study (cannabis cessation therapies), studies included a wide range of interventions and considerable variation in study populations and outcomes. A brief summary of the key findings for each study was presented and narrative synthesis used to summarise results for each

  17. [Development of a rapid test kit for antibody to HIV by nano immunomagnetic lateral flow method].

    Science.gov (United States)

    Yang, Fa-qing; Lee, Tony; Wang, Chao-nan; Sun, Shu-ye; Li, Shan-shan; Tian, Hui

    2010-06-01

    To develop a rapid test kit for antibody to HIV by nano immunomagnetic lateral flow method. A rapid test kit was developed by conjugation of the HIV antigen gp41 and gp36 to 200nm super paramagnetic particles by carbodiimide (EDC) and coating of the HIV antigen gp41 and gp36 to nitrocellulose membrane. Then the kit was evaluated with serials of experiments. The kit was qualified with examination of national reference panel of anti-HIV antibody for colloidal gold diagnostic kit. The sensitivity was 100% by tested with 20 HIV antibody positive sera, the specificity was 98.5% by tested with 600 HIV antibody negative sera, respectively. The stability of the kit was over 12 month by storage at room temperature. A diagnostic kit for antibody to HIV was developed with the advantages of convenience, rapid test, good stability and point of care.

  18. Rapid culture-based methods for drug-resistance detection in Mycobacterium tuberculosis.

    Science.gov (United States)

    Palomino, Juan Carlos; Martin, Anandi; Von Groll, Andrea; Portaels, Francoise

    2008-10-01

    Tuberculosis still represents a major public health problem, especially in low-resource countries where the burden of the disease is more important. Multidrug-resistant and extensively drug drug-resistant tuberculosis constitute serious problems for the efficient control of the disease stressing the need to investigate resistance to first- and second-line drugs. Conventional methods for detecting drug-resistance in Mycobacterium tuberculosis are slow and cumbersome. The most commonly used proportion method on Löwenstein-Jensen medium or Middlebrook agar requires a minimum of 3-4 weeks to produce results. Several new approaches have been proposed in the last years for the rapid and timely detection of drug-resistance in tuberculosis. This review will address phenotypic culture-based methods for rapid drug susceptibility testing in M. tuberculosis.

  19. Determination of rapid chlorination rate constants by a stopped-flow spectrophotometric competition kinetics method.

    Science.gov (United States)

    Song, Dean; Liu, Huijuan; Qiang, Zhimin; Qu, Jiuhui

    2014-05-15

    Free chlorine is extensively used for water and wastewater disinfection nowadays. However, it still remains a big challenge to determine the rate constants of rapid chlorination reactions although competition kinetics and stopped-flow spectrophotometric (SFS) methods have been employed individually to investigate fast reaction kinetics. In this work, we proposed an SFS competition kinetics method to determine the rapid chlorination rate constants by using a common colorimetric reagent, N,N-diethyl-p-phenylenediamine (DPD), as a reference probe. A kinetic equation was first derived to estimate the reaction rate constant of DPD towards chlorine under a given pH and temperature condition. Then, on that basis, an SFS competition kinetics method was proposed to determine directly the chlorination rate constants of several representative compounds including tetracycline, ammonia, and four α-amino acids. Although Cl2O is more reactive than HOCl, its contribution to the overall chlorination kinetics of the test compounds could be neglected in this study. Finally, the developed method was validated through comparing the experimentally measured chlorination rate constants of the selected compounds with those obtained or calculated from literature and analyzing with Taft's correlation as well. This study demonstrates that the SFS competition kinetics method can measure the chlorination rate constants of a test compound rapidly and accurately. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Rapid Column Extraction Method for Actinides and Sr-89/90 in Water Samples

    Energy Technology Data Exchange (ETDEWEB)

    MAXWELL III, SHERROD L.

    2005-06-15

    The SRS Environmental Laboratory analyzes water samples for environmental monitoring, including river water and ground water samples. A new, faster actinide and strontium 89/90 separation method has been developed and implemented to improve productivity, reduce labor costs and add capacity to this laboratory. This method uses stacked TEVA Resin{reg_sign}, TRU Resin{reg_sign} and Sr-Resin{reg_sign} cartridges from Eichrom Technologies (Darien, IL, USA) that allows the rapid separation of plutonium (Pu), neptunium (Np), uranium (U), americium (Am), curium (Cm) and thorium (Th) using a single multi-stage column combined with alpha spectrometry. By using vacuum box cartridge technology with rapid flow rates, sample preparation time is minimized. The method can be used for routine analysis or as a rapid method for emergency preparedness. Thorium and curium are often analyzed separately due to the interference of the daughter of Th-229 tracer, actinium (Ac)-225, on curium isotopes when measured by alpha spectrometry. This new method also adds a separation step using DGA Resin{reg_sign}, (Diglycolamide Resin, Eichrom Technologies) to remove Ac-225 and allow the separation and analysis of thorium isotopes and curium isotopes at the same time.

  1. Mycobacteria mobility shift assay: a method for the rapid identification of Mycobacterium tuberculosis and nontuberculous mycobacteria

    Directory of Open Access Journals (Sweden)

    Letícia Muraro Wildner

    2014-06-01

    Full Text Available The identification of mycobacteria is essential because tuberculosis (TB and mycobacteriosis are clinically indistinguishable and require different therapeutic regimens. The traditional phenotypic method is time consuming and may last up to 60 days. Indeed, rapid, affordable, specific and easy-to-perform identification methods are needed. We have previously described a polymerase chain reaction-based method called a mycobacteria mobility shift assay (MMSA that was designed for Mycobacterium tuberculosis complex (MTC and nontuberculous mycobacteria (NTM species identification. The aim of this study was to assess the MMSA for the identification of MTC and NTM clinical isolates and to compare its performance with that of the PRA-hsp65 method. A total of 204 clinical isolates (102 NTM and 102 MTC were identified by the MMSA and PRA-hsp65. For isolates for which these methods gave discordant results, definitive species identification was obtained by sequencing fragments of the 16S rRNA and hsp65 genes. Both methods correctly identified all MTC isolates. Among the NTM isolates, the MMSA alone assigned 94 (92.2% to a complex or species, whereas the PRA-hsp65 method assigned 100% to a species. A 91.5% agreement was observed for the 94 NTM isolates identified by both methods. The MMSA provided correct identification for 96.8% of the NTM isolates compared with 94.7% for PRA-hsp65. The MMSA is a suitable auxiliary method for routine use for the rapid identification of mycobacteria.

  2. Adapting and Evaluating a Rapid, Low-Cost Method to Enumerate Flies in the Household Setting.

    Science.gov (United States)

    Wolfe, Marlene K; Dentz, Holly N; Achando, Beryl; Mureithi, MaryAnne; Wolfe, Tim; Null, Clair; Pickering, Amy J

    2017-02-08

    Diarrhea is a leading cause of death among children under 5 years of age worldwide. Flies are important vectors of diarrheal pathogens in settings lacking networked sanitation services. There is no standardized method for measuring fly density in households; many methods are cumbersome and unvalidated. We adapted a rapid, low-cost fly enumeration technique previously developed for industrial settings, the Scudder fly grill, for field use in household settings. We evaluated its performance in comparison to a sticky tape fly trapping method at latrine and food preparation areas among households in rural Kenya. The grill method was more sensitive; it detected the presence of any flies at 80% (433/543) of sampling locations versus 64% (348/543) of locations by the sticky tape. We found poor concordance between the two methods, suggesting that standardizing protocols is important for comparison of fly densities between studies. Fly species identification was feasible with both methods; however, the sticky tape trap allowed for more nuanced identification. Both methods detected a greater presence of bottle flies near latrines compared with food preparation areas (P < 0.01). The grill method detected more flies at the food preparation area compared with near the latrine (P = 0.014) while the sticky tape method detected no difference. We recommend the Scudder grill as a sensitive fly enumeration tool that is rapid and low cost to implement. © The American Society of Tropical Medicine and Hygiene.

  3. Rapid and effective DNA extraction method with bead grinding for a large amount of fungal DNA.

    Science.gov (United States)

    Watanabe, M; Lee, K; Goto, K; Kumagai, S; Sugita-Konishi, Y; Hara-Kudo, Y

    2010-06-01

    To identify a rapid method for extracting a large amount of DNA from fungi associated with food hygiene, extraction methods were compared using fungal pellets formed rapidly in liquid media. Combinations of physical and chemical methods or commercial kits were evaluated with 3 species of yeast, 10 species of ascomycetous molds, and 4 species of zygomycetous molds. Bead grinding was the physical method, followed by chemical methods involving sodium dodecyl sulfate (SDS), cetyl trimethyl ammonium bromide (CTAB), and benzyl chloride and two commercial kits. Quantity was calculated by UV absorbance at 260 nm, quality was determined by the ratio of UV absorbance at 260 and 280 nm, and gene amplifications and electrophoresis profiles of whole genomes were analyzed. Bead grinding with the SDS method was the most effective for DNA extraction for yeasts and ascomycetous molds, and bead grinding with the CTAB method was most effective with zygomycetous molds. For both groups of molds, bead grinding with the CTAB method was the best approach for DNA extraction. Because this combination also is relatively effective for yeasts, it can be used to extract a large amount of DNA from a wide range of fungi. The DNA extraction methods are useful for developing gene indexes to identify fungi with molecular techniques, such as DNA fingerprinting.

  4. Mycobacteria mobility shift assay: a method for the rapid identification of Mycobacterium tuberculosis and nontuberculous mycobacteria.

    Science.gov (United States)

    Wildner, Letícia Muraro; Bazzo, Maria Luiza; Liedke, Susie Coutinho; Nogueira, Christiane Lourenço; Segat, Gabriela; Senna, Simone Gonçalves; Schlindwein, Aline Daiane; Oliveira, Jaquelline Germano de; Rovaris, Darcita B; Bonjardim, Claudio A; Kroon, Erna G; Ferreira, Paulo C P

    2014-06-01

    The identification of mycobacteria is essential because tuberculosis (TB) and mycobacteriosis are clinically indistinguishable and require different therapeutic regimens. The traditional phenotypic method is time consuming and may last up to 60 days. Indeed, rapid, affordable, specific and easy-to-perform identification methods are needed. We have previously described a polymerase chain reaction-based method called a mycobacteria mobility shift assay (MMSA) that was designed for Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) species identification. The aim of this study was to assess the MMSA for the identification of MTC and NTM clinical isolates and to compare its performance with that of the PRA-hsp65 method. A total of 204 clinical isolates (102 NTM and 102 MTC) were identified by the MMSA and PRA-hsp65. For isolates for which these methods gave discordant results, definitive species identification was obtained by sequencing fragments of the 16S rRNA and hsp65 genes. Both methods correctly identified all MTC isolates. Among the NTM isolates, the MMSA alone assigned 94 (92.2%) to a complex or species, whereas the PRA-hsp65 method assigned 100% to a species. A 91.5% agreement was observed for the 94 NTM isolates identified by both methods. The MMSA provided correct identification for 96.8% of the NTM isolates compared with 94.7% for PRA-hsp65. The MMSA is a suitable auxiliary method for routine use for the rapid identification of mycobacteria.

  5. A rapid chromatographic method for quality control of technetium-99m-bicisate.

    Science.gov (United States)

    Amin, K C; Saha, G B; Go, R T

    1997-03-01

    The purpose of this work was to develop a simple and rapid method to determine the radiochemical purity of 99mTc-bicisate. A rapid paper chromatographic (PC) method was developed to determine the radiochemical purity of 99mTc-bicisate and compare the results with those of the manufacturer's recommended method. The present PC method included Whatman 3MM paper as the solid phase and ethyl acetate as the solvent. The time for chromatography by this technique was 4-5 min compared to about 23 min by the manufacturer's method. The Rf value of 99mTc-bicisate (Rf = 0.9-1.0) was widely different from those of 99mTcO4- and reduced 99mTc (Rf = 0.0 for both) so the chromatographic strip after development could be readily cut into two segments, in order to determine the labeling yield. No significant difference in labeling yields was found between the present method and the manufacturer's method. The PC method using Whatman 3MM paper and ethyl acetate is a simple and fast technique to determine the radiochemical purity of 99mTc-bicisate and may be substituted for the manufacturer's recommended method to save time.

  6. Egzemplifikacja Tomograficznej Metody Do Oceny Jakości Połączeń Spawanych Ze Stopu EN 5754 H22

    Directory of Open Access Journals (Sweden)

    Błachnio Józef

    2016-12-01

    Full Text Available Jakość połączeń spawanych zależna jest od wielu czynników. Przedmiotowe normy określają warunki techniczne oceny jakościowej spawów, co stanowi podstawę do uznania połączenia za zgodne z wymaganiami lub wadliwe. W praktyce wykonanie połączenia spawanego całkowicie pozbawionego wad jest trudne. Do kontroli wewnętrznej struktury połączenia wykorzystać można nieniszczącą metodę z zastosowaniem przemysłowego tomografu komputerowego. Ta nowoczesna metoda diagnozowania łączy badanie rentgenowskie z zaawansowaną technologią komputerową. Podstawową zaletą tomografii komputerowej jest badanie obiektów w trzech wymiarach oraz możliwość wykonania rekonstrukcji przestrzennych. Celem artykułu jest omówienie wykorzystania tej metody do oceny jakości połączeń spawanych stopów aluminium. Możliwości tomografii komputerowej przedstawiono na przykładzie próbek spawalniczych wykonanych metodą TIG przy różnych parametrach procesu. Dokonano analizy jakości próbek wykazujących najmniejsze i największe zewnętrzne, jak również wewnętrzne wady spawalnicze.

  7. Visual and colorimetric methods for rapid determination of total tannins in vegetable raw materials

    Directory of Open Access Journals (Sweden)

    S. P. Kalinkina

    2016-01-01

    Full Text Available The article is dedicated to the development of rapid colorimetric method for determining the amount of tannins in aqueous extracts of vegetable raw materials. The sorption-based colorimetric test is determining sorption tannins polyurethane foam, impregnated of FeCl3, receiving on its surface painted in black and green color of the reaction products and the determination of their in sorbent matrix. Selectivity is achieved by determining the tannins specific interaction of polyphenols with iron ions (III. The conditions of sorption-colorimetric method: the concentration of ferric chloride (III, impregnated in the polyurethane foam; sorbent mass in the analytical cartridge; degree of loading his agent; the contact time of the phases. color scales have been developed for the visual determination of the amount of tannins in terms of gallic acid. Spend a digitized image obtained scales using computer program “Sorbfil TLC”, excluding a subjective assessment of the intensity of the color scale of the test. The results obtained determine the amount of tannins in aqueous extracts of vegetable raw rapid method using tablets and analytical cartridges. The results of the test determination of tannins with visual and densitometric analytical signal registration are compared to known methods. Spend a metrological evaluation of the results of determining the amount of tannins sorption rapid colorimetric methods. Time visual and densitometric rapid determination of tannins, taking into account the sample preparation is 25–30 minutes, the relative error does not exceed 28 %. The developed test methods for quantifying the content of tannins allow to exclude the use of sophisticated analytical equipment, carry out the analysis in non-laboratory conditions do not require highly skilled personnel.

  8. A rapid method for soil cement design : Louisiana slope value method : part II : evaluation.

    Science.gov (United States)

    1966-05-01

    This report is an evaluation of the recently developed "Louisiana Slope Value Method". : The conclusion drawn are based on data from 637 separate samples representing nearly all major soil groups in Louisiana that are suitable for cement stabilizatio...

  9. Melting Temperature Mapping Method: A Novel Method for Rapid Identification of Unknown Pathogenic Microorganisms within Three Hours of Sample Collection.

    Science.gov (United States)

    Niimi, Hideki; Ueno, Tomohiro; Hayashi, Shirou; Abe, Akihito; Tsurue, Takahiro; Mori, Masashi; Tabata, Homare; Minami, Hiroshi; Goto, Michihiko; Akiyama, Makoto; Yamamoto, Yoshihiro; Saito, Shigeru; Kitajima, Isao

    2015-07-28

    Acquiring the earliest possible identification of pathogenic microorganisms is critical for selecting the appropriate antimicrobial therapy in infected patients. We herein report the novel "melting temperature (Tm) mapping method" for rapidly identifying the dominant bacteria in a clinical sample from sterile sites. Employing only seven primer sets, more than 100 bacterial species can be identified. In particular, using the Difference Value, it is possible to identify samples suitable for Tm mapping identification. Moreover, this method can be used to rapidly diagnose the absence of bacteria in clinical samples. We tested the Tm mapping method using 200 whole blood samples obtained from patients with suspected sepsis, 85% (171/200) of which matched the culture results based on the detection level. A total of 130 samples were negative according to the Tm mapping method, 98% (128/130) of which were also negative based on the culture method. Meanwhile, 70 samples were positive according to the Tm mapping method, and of the 59 suitable for identification, 100% (59/59) exhibited a "match" or "broad match" with the culture or sequencing results. These findings were obtained within three hours of whole blood collection. The Tm mapping method is therefore useful for identifying infectious diseases requiring prompt treatment.

  10. 3D virtual human rapid modeling method based on top-down modeling mechanism

    Directory of Open Access Journals (Sweden)

    LI Taotao

    2017-01-01

    Full Text Available Aiming to satisfy the vast custom-made character demand of 3D virtual human and the rapid modeling in the field of 3D virtual reality, a new virtual human top-down rapid modeling method is put for-ward in this paper based on the systematic analysis of the current situation and shortage of the virtual hu-man modeling technology. After the top-level realization of virtual human hierarchical structure frame de-sign, modular expression of the virtual human and parameter design for each module is achieved gradu-al-level downwards. While the relationship of connectors and mapping restraints among different modules is established, the definition of the size and texture parameter is also completed. Standardized process is meanwhile produced to support and adapt the virtual human top-down rapid modeling practice operation. Finally, the modeling application, which takes a Chinese captain character as an example, is carried out to validate the virtual human rapid modeling method based on top-down modeling mechanism. The result demonstrates high modelling efficiency and provides one new concept for 3D virtual human geometric mod-eling and texture modeling.

  11. Development of a loop-mediated isothermal amplification method for rapid detection of pigeon circovirus.

    Science.gov (United States)

    Tsai, Shinn Shyong; Chang, Yeng Ling; Huang, Yen Li; Liu, Hung Jen; Ke, Guan Ming; Chiou, Chwei Jang; Hsieh, Yao Ching; Chang, Tsung Chou; Cheng, Li Ting; Chuang, Kuo Pin

    2014-05-01

    There are no effective antiviral treatments for pigeon circovirus (PiCV); thus, rapid diagnosis is critical for effective control of the disease caused by this virus. The recent development of a novel LAMP technique that amplifies nucleic acids rapidly with high specificity and sensitivity under isothermal conditions has overcome some of the deficiencies of nucleic-acid-based diagnostic tests. We established a LAMP method for rapid detection of PiCV using two pairs of primers that were designed from PiCV and compared its sensitivity and specificity with that of PCR. Amplification by LAMP was optimal at 63 °C for 60 min. The detection limit was nearly 0.5 pg of PiCV DNA, making it ten times more sensitive than PCR. There was no cross-reaction with porcine circovirus type 2 (PCV2), pigeon Trichomonas gallinae, or pigeon herpesvirus (PHV) under the same conditions. The assay also successfully detected the pathogen DNA in the tissues of infected pigeons. This is the first report indicating that LAMP is a valuable, rapid method of detecting PiCV with high sensitivity and specificity.

  12. A new alginate-based rapid method for determining coliforms in milk.

    Science.gov (United States)

    Chang, Su-sen; Gray, Peter M; Woo, Gun-Jo; Kang, Dong-Hyun

    2003-11-01

    A new rapid method for monitoring coliforms was developed on the basis of the instant gelling effects of alginate and calcium. The effectiveness of this new method in the detection of coliforms was evaluated. Tests involving Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, total coliforms in milk, cold-injured coliforms, and total coliforms in raw milk were carried out. The bacterial samples were diluted in 0.2% peptone water containing 90 mM CaCl2 and added into test tubes containing modified purple broth base medium. Coliform concentrations were determined on the basis of the time of color change and gas production in the alginate tubes. All results obtained by the alginate method correlated strongly with those obtained by the conventional violet red bile agar (VRBA) plating method. The alginate method reduced detection time by 12 to 14 h compared with the conventional VRBA plating method. The alginate method can be applied in field studies more easily than melted-agar systems can. The results of this study indicate that the alginate method is an accurate, rapid, simple, and economical way to monitor and estimate concentrations of total coliforms in food.

  13. New method for vitrifying water and other liquids by rapid cooling of their aerosols

    Science.gov (United States)

    Mayer, Erwin

    1985-07-01

    A method for the vitrification of pure liquid water and dilute aqueous solutions is described which is the only one without a liquid cryomedium for heat transfer: rapid cooling of aqueous aerosol droplets on a solid cryoplate. This method is not limited to water and aqueous solutions, but can be used for the vitrification of any liquid aerosol, the only impurity being some codeposited vapor. The method can be applied in diverse fields such as cryobiology, cryomicroscopy, and low-temperature spectroscopy of water and dilute aqueous solutions to avoid the formation of crystalline ice.

  14. Rapid simulation of electromagnetic telemetry using an axisymmetric semianalytical finite element method

    Science.gov (United States)

    Chen, Jiefu; Zeng, Shubin; Dong, Qiuzhao; Huang, Yueqin

    2017-02-01

    An axisymmetric semianalytical finite element method is proposed and employed for rapid simulations of electromagnetic telemetry in layered underground formation. In this method, the layered media is decomposed into several subdomains and the interfaces between subdomains are discretized by conventional finite elements. Then a Riccati equation based high precision integration scheme is applied to exploit the homogeneity along the vertical direction in each layer. This semianalytical finite element scheme is very efficient in modeling electromagnetic telemetry in layered formation. Numerical examples as well as a field case with water based mud as drilling fluid are given to demonstrate the validity and effectiveness of this method.

  15. RAPID METHOD FOR PLUTONIUM, AMERICIUM AND CURIUM IN VERY LARGE SOIL SAMPLES

    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, S

    2007-01-08

    The analysis of actinides in environmental soil and sediment samples is very important for environmental monitoring. There is a need to measure actinide isotopes with very low detection limits. A new, rapid actinide separation method has been developed and implemented that allows the measurement of plutonium, americium and curium isotopes in very large soil samples (100-200 g) with high chemical recoveries and effective removal of matrix interferences. This method uses stacked TEVA Resin{reg_sign}, TRU Resin{reg_sign} and DGA-Resin{reg_sign} cartridges from Eichrom Technologies (Darien, IL, USA) that allows the rapid separation of plutonium (Pu), americium (Am), and curium (Cm) using a single multistage column combined with alpha spectrometry. The method combines an acid leach step and innovative matrix removal using cerium fluoride precipitation to remove the difficult soil matrix. This method is unique in that it provides high tracer recoveries and effective removal of interferences with small extraction chromatography columns instead of large ion exchange resin columns that generate large amounts of acid waste. By using vacuum box cartridge technology with rapid flow rates, sample preparation time is minimized.

  16. A novel sample preparation method using rapid nonheated saponification method for the determination of cholesterol in emulsified foods.

    Science.gov (United States)

    Jeong, In-Seek; Kwak, Byung-Man; Ahn, Jang-Hyuk; Leem, Donggil; Yoon, Taehyung; Yoon, Changyong; Jeong, Jayoung; Park, Jung-Min; Kim, Jin-Man

    2012-10-01

    In this study, nonheated saponification was employed as a novel, rapid, and easy sample preparation method for the determination of cholesterol in emulsified foods. Cholesterol content was analyzed using gas chromatography with a flame ionization detector (GC-FID). The cholesterol extraction method was optimized for maximum recovery from baby food and infant formula. Under these conditions, the optimum extraction solvent was 10 mL ethyl ether per 1 to 2 g sample, and the saponification solution was 0.2 mL KOH in methanol. The cholesterol content in the products was determined to be within the certified range of certified reference materials (CRMs), NIST SRM 1544 and SRM 1849. The results of the recovery test performed using spiked materials were in the range of 98.24% to 99.45% with an relative standard devitation (RSD) between 0.83% and 1.61%. This method could be used to reduce sample pretreatment time and is expected to provide an accurate determination of cholesterol in emulsified food matrices such as infant formula and baby food. A novel, rapid, and easy sample preparation method using nonheated saponification was developed for cholesterol detection in emulsified foods. Recovery tests of CRMs were satisfactory, and the recoveries of spiked materials were accurate and precise. This method was effective and decreased the time required for analysis by 5-fold compared to the official method. © 2012 Institute of Food Technologists®

  17. A Rapid and Economical Method for Efficient DNA Extraction from Diverse Soils Suitable for Metagenomic Applications.

    Directory of Open Access Journals (Sweden)

    Selvaraju Gayathri Devi

    Full Text Available A rapid, cost effective method of metagenomic DNA extraction from soil is a useful tool for environmental microbiology. The present work describes an improved method of DNA extraction namely "powdered glass method" from diverse soils. The method involves the use of sterile glass powder for cell lysis followed by addition of 1% powdered activated charcoal (PAC as purifying agent to remove humic substances. The method yielded substantial DNA (5.87 ± 0.04 μg/g of soil with high purity (A260/280: 1.76 ± 0.05 and reduced humic substances (A340: 0.047 ± 0.03. The quality of the extracted DNA was compared against five different methods based on 16S rDNA PCR amplification, BamHI digestion and validated using quantitative PCR. The digested DNA was used for a metagenomic library construction with the transformation efficiency of 4 X 106 CFU mL-1. Besides providing rapid, efficient and economical extraction of metgenomic DNA from diverse soils, this method's applicability is also demonstrated for cultivated organisms (Gram positive B. subtilis NRRL-B-201, Gram negative E. coli MTCC40, and a microalgae C. sorokiniana UTEX#1666.

  18. A Rapid and Economical Method for Efficient DNA Extraction from Diverse Soils Suitable for Metagenomic Applications.

    Science.gov (United States)

    Devi, Selvaraju Gayathri; Fathima, Anwar Aliya; Radha, Sudhakar; Arunraj, Rex; Curtis, Wayne R; Ramya, Mohandass

    2015-01-01

    A rapid, cost effective method of metagenomic DNA extraction from soil is a useful tool for environmental microbiology. The present work describes an improved method of DNA extraction namely "powdered glass method" from diverse soils. The method involves the use of sterile glass powder for cell lysis followed by addition of 1% powdered activated charcoal (PAC) as purifying agent to remove humic substances. The method yielded substantial DNA (5.87 ± 0.04 μg/g of soil) with high purity (A260/280: 1.76 ± 0.05) and reduced humic substances (A340: 0.047 ± 0.03). The quality of the extracted DNA was compared against five different methods based on 16S rDNA PCR amplification, BamHI digestion and validated using quantitative PCR. The digested DNA was used for a metagenomic library construction with the transformation efficiency of 4 X 106 CFU mL-1. Besides providing rapid, efficient and economical extraction of metgenomic DNA from diverse soils, this method's applicability is also demonstrated for cultivated organisms (Gram positive B. subtilis NRRL-B-201, Gram negative E. coli MTCC40, and a microalgae C. sorokiniana UTEX#1666).

  19. Simple and rapid method for the isolation of forskolin from Coleus forskohlii by charcoal column chromatography.

    Science.gov (United States)

    Saleem, A M; Dhasan, P B; Rafiullah, M R M

    2006-01-06

    A simple, safe, rapid and economical method was developed for the isolation of high-purity forskolin from Coleus forskohlii roots using activated charcoal as an adsorbent in a column. The elution was carried out under reduced pressure to make the process rapid. Activated charcoal acted as a reversed phase adsorbent and allowed elution of forskolin without much impurities. The residue, obtained from the eluate was purified and crystallized using different solvent mixtures to obtain pure forskolin. The forskolin isolated was analyzed and characterized by UV, IR, RP-HPLC, electrospray ionization MS, 1H NMR and 13C NMR. The yield was 0.097% w/w (RSD 5.6%). The purity was 96.9% w/w (RSD 0.3%) as determined by RP-HPLC. The present method enables researchers to produce high-purity forskolin in their labs by using common chemicals.

  20. Evaluation of methods for rapid determination of freezing point of aviation fuels

    Science.gov (United States)

    Mathiprakasam, B.

    1982-01-01

    Methods for identification of the more promising concepts for the development of a portable instrument to rapidly determine the freezing point of aviation fuels are described. The evaluation process consisted of: (1) collection of information on techniques previously used for the determination of the freezing point, (2) screening and selection of these techniques for further evaluation of their suitability in a portable unit for rapid measurement, and (3) an extensive experimental evaluation of the selected techniques and a final selection of the most promising technique. Test apparatuses employing differential thermal analysis and the change in optical transparency during phase change were evaluated and tested. A technique similar to differential thermal analysis using no reference fuel was investigated. In this method, the freezing point was obtained by digitizing the data and locating the point of inflection. Results obtained using this technique compare well with those obtained elsewhere using different techniques. A conceptual design of a portable instrument incorporating this technique is presented.

  1. Novel methods for improving rapid paper-based protein assays with gold nanoparticle detection

    OpenAIRE

    Lama, Lara

    2017-01-01

    This thesis describes methods for improving sensitivity in rapid singleplex and multiplex microarray assays. The assays utilize the optical characteristics of colloidal gold nanoparticles for the colorimetric detection of proteins. Multiplexed detection in sandwich immunoassays is limited by cross-reactivity between different detection antibodies. The cross-reactivity between antibodies can contribute to increased background noise - decreasing the Limit-of-Detection of the assay - or generate...

  2. Rapid direct methods for enumeration of specific, active bacteria in water and biofilms

    Science.gov (United States)

    McFeters, G. A.; Pyle, B. H.; Lisle, J. T.; Broadaway, S. C.

    1999-01-01

    Conventional methods for detecting indicator and pathogenic bacteria in water may underestimate the actual population due to sublethal environmental injury, inability of the target bacteria to take up nutrients and other physiological factors which reduce bacterial culturability. Rapid and direct methods are needed to more accurately detect and enumerate active bacteria. Such a methodological advance would provide greater sensitivity in assessing the microbiological safety of water and food. The principle goal of this presentation is to describe novel approaches we have formulated for the rapid and simultaneous detection of bacteria plus the determination of their physiological activity in water and other environmental samples. The present version of our method involves the concentration of organisms by membrane filtration or immunomagnetic separation and combines an intracellular fluorochrome (CTC) for assessment of respiratory activity plus fluorescent-labelled antibody detection of specific bacteria. This approach has also been successfully used to demonstrate spatial and temporal heterogeneities of physiological activities in biofilms when coupled with cryosectioning. Candidate physiological stains include those capable of determining respiratory activity, membrane potential, membrane integrity, growth rate and cellular enzymatic activities. Results obtained thus far indicate that immunomagnetic separation can provide a high degree of sensitivity in the recovery of seeded target bacteria (Escherichia coli O157:H7) in water and hamburger. The captured and stained target bacteria are then enumerated by either conventional fluorescence microscopy or ChemScan(R), a new instrument that is very sensitive and rapid. The ChemScan(R) laser scanning instrument (Chemunex, Paris, France) provides the detection of individual fluorescently labelled bacterial cells using three emission channels in less than 5 min. A high degree of correlation has been demonstrated between

  3. New multiplex PCR methods for rapid screening of genetically modified organisms in foods

    OpenAIRE

    Nelly eDatukishvili; Tamara eKutateladze; Inga eGabriadze; Kakha eBitskinashvili; Boris eVishnepolsky

    2015-01-01

    We present novel multiplex PCR methods for rapid and reliable screening of genetically modified organisms (GMOs). New designed PCR primers targeting four frequently used GMO specific sequences permitted identification of new DNA markers, in particular 141 bp fragment of cauliflower mosaic virus (CaMV) 35S promoter, 224 bp fragment of Agrobacterium tumefaciens nopaline synthase (NOS) terminator, 256 bp fragment of 5-enolppyruvylshikimate-phosphate synthase (epsps) gene and 258 bp fragment of C...

  4. A low complexity rapid molecular method for detection of Clostridium difficile in stool.

    Directory of Open Access Journals (Sweden)

    Cathal J McElgunn

    Full Text Available Here we describe a method for the detection of Clostridium difficile from stool using a novel low-complexity and rapid extraction process called Heat Elution (HE. The HE method is two-step and takes just 10 minutes, no specialist instruments are required and there is minimal hands-on time. A test method using HE was developed in conjunction with Loop-mediated Isothermal Amplification (LAMP combined with the real-time bioluminescent reporter system known as BART targeting the toxin B gene (tcdB. The HE-LAMP-BART method was evaluated in a pilot study on clinical fecal samples (tcdB(+, n = 111; tcdB(-, n= 107. The HE-LAMP-BART method showed 95.5% sensitivity and 100% specificity against a gold standard reference method using cytotoxigenic culture and also a silica-based robotic extraction followed by tcdB PCR to control for storage. From sample to result, the HE-LAMP-BART method typically took 50 minutes, whereas the PCR method took >2.5 hours. In a further study (tcdB(+, n = 47; tcdB(-, n= 28 HE-LAMP-BART was compared to an alternative commercially available LAMP-based method, Illumigene (Meridian Bioscience, OH, and yielded 87.2% sensitivity and 100% specificity for the HE-LAMP-BART method compared to 76.6% and 100%, respectively, for Illumigene against the reference method. A subset of 27 samples (tcdB(+, n = 25; tcdB(-, n= 2 were further compared between HE-LAMP-BART, Illumigene, GeneXpert (Cepheid, Sunnyvale, CA and RIDA®QUICK C. difficile Toxin A/B lateral flow rapid test (R-Biopharm, Darmstadt, Germany resulting in sensitivities of HE-LAMP-BART 92%, Illumigene 72% GeneXpert 96% and RIDAQuick 76% against the reference method. The HE-LAMP-BART method offers the advantages of molecular based approaches without the cost and complexity usually associated with molecular tests. Further, the rapid time-to-result and simple protocol means the method can be applied away from the centralized laboratory settings.

  5. Conventional rapid latex agglutination in estimation of von Willebrand factor: method revisited and potential clinical applications.

    Science.gov (United States)

    Mahat, Marianor; Abdullah, Wan Zaidah; Hussin, Che Maraina Che

    2014-01-01

    Measurement of von Willebrand factor antigen (VWF : Ag) levels is usually performed in a specialised laboratory which limits its application in routine clinical practice. So far, no commercial rapid test kit is available for VWF : Ag estimation. This paper discusses the technical aspect of latex agglutination method which was established to suit the purpose of estimating von Willebrand factor (VWF) levels in the plasma sample. The latex agglutination test can be performed qualitatively and semiquantitatively. Reproducibility, stability, linearity, limit of detection, interference, and method comparison studies were conducted to evaluate the performance of this test. Semiquantitative latex agglutination test was strongly correlated with the reference immunoturbidimetric assay (Spearman's rho = 0.946, P agglutination test and the reference assay. Using the scoring system for the rapid latex test, no agglutination is with 0% VWF : Ag (control negative), 1+ reaction is equivalent to 150% VWF : Ag (when comparing with immunoturbidimetric assay). The findings from evaluation studies suggest that latex agglutination method is suitable to be used as a rapid test kit for the estimation of VWF : Ag levels in various clinical conditions associated with high levels and low levels of VWF : Ag.

  6. A direct and rapid method to determine cyanide in urine by capillary electrophoresis.

    Science.gov (United States)

    Zhang, Qiyang; Maddukuri, Naveen; Gong, Maojun

    2015-10-02

    Cyanides are poisonous chemicals that widely exist in nature and industrial processes as well as accidental fires. Rapid and accurate determination of cyanide exposure would facilitate forensic investigation, medical diagnosis, and chronic cyanide monitoring. Here, a rapid and direct method was developed for the determination of cyanide ions in urinary samples. This technique was based on an integrated capillary electrophoresis system coupled with laser-induced fluorescence (LIF) detection. Cyanide ions were derivatized with naphthalene-2,3-dicarboxaldehyde (NDA) and a primary amine (glycine) for LIF detection. Three separate reagents, NDA, glycine, and cyanide sample, were mixed online, which secured uniform conditions between samples for cyanide derivatization and reduced the risk of precipitation formation of mixtures. Conditions were optimized; the derivatization was completed in 2-4min, and the separation was observed in 25s. The limit of detection (LOD) was 4.0nM at 3-fold signal-to-noise ratio for standard cyanide in buffer. The cyanide levels in urine samples from smokers and non-smokers were determined by using the method of standard addition, which demonstrated significant difference of cyanide levels in urinary samples from the two groups of people. The developed method was rapid and accurate, and is anticipated to be applicable to cyanide detection in waste water with appropriate modification. Published by Elsevier B.V.

  7. GSMA: Gene Set Matrix Analysis, An Automated Method for Rapid Hypothesis Testing of Gene Expression Data

    Directory of Open Access Journals (Sweden)

    Chris Cheadle

    2007-01-01

    Full Text Available Background: Microarray technology has become highly valuable for identifying complex global changes in gene expression patterns. The assignment of functional information to these complex patterns remains a challenging task in effectively interpreting data and correlating results from across experiments, projects and laboratories. Methods which allow the rapid and robust evaluation of multiple functional hypotheses increase the power of individual researchers to data mine gene expression data more efficiently.Results: We have developed (gene set matrix analysis GSMA as a useful method for the rapid testing of group-wise up- or downregulation of gene expression simultaneously for multiple lists of genes (gene sets against entire distributions of gene expression changes (datasets for single or multiple experiments. The utility of GSMA lies in its flexibility to rapidly poll gene sets related by known biological function or as designated solely by the end-user against large numbers of datasets simultaneously.Conclusions: GSMA provides a simple and straightforward method for hypothesis testing in which genes are tested by groups across multiple datasets for patterns of expression enrichment.

  8. Interactive Rapid Dose Assessment Model (IRDAM): reactor-accident assessment methods. Vol. 2

    Energy Technology Data Exchange (ETDEWEB)

    Poeton, R.W.; Moeller, M.P.; Laughlin, G.J.; Desrosiers, A.E.

    1983-05-01

    As part of the continuing emphasis on emergency preparedness, the US Nuclear Regulatory Commission (NRC) sponsored the development of a rapid dose assessment system by Pacific Northwest Laboratory (PNL). This system, the Interactive Rapid Dose Assessment Model (IRDAM) is a micro-computer based program for rapidly assessing the radiological impact of accidents at nuclear power plants. This document describes the technical bases for IRDAM including methods, models and assumptions used in calculations. IRDAM calculates whole body (5-cm depth) and infant thyroid doses at six fixed downwind distances between 500 and 20,000 meters. Radionuclides considered primarily consist of noble gases and radioiodines. In order to provide a rapid assessment capability consistent with the capacity of the Osborne-1 computer, certain simplifying approximations and assumptions are made. These are described, along with default values (assumptions used in the absence of specific input) in the text of this document. Two companion volumes to this one provide additional information on IRDAM. The user's Guide (NUREG/CR-3012, Volume 1) describes the setup and operation of equipment necessary to run IRDAM. Scenarios for Comparing Dose Assessment Models (NUREG/CR-3012, Volume 3) provides the results of calculations made by IRDAM and other models for specific accident scenarios.

  9. Comparison of concentration methods for rapid detection of hookworm ova in wastewater matrices using quantitative PCR.

    Science.gov (United States)

    Gyawali, P; Ahmed, W; Jagals, P; Sidhu, J P S; Toze, S

    2015-12-01

    Hookworm infection contributes around 700 million infections worldwide especially in developing nations due to increased use of wastewater for crop production. The effective recovery of hookworm ova from wastewater matrices is difficult due to their low concentrations and heterogeneous distribution. In this study, we compared the recovery rates of (i) four rapid hookworm ova concentration methods from municipal wastewater, and (ii) two concentration methods from sludge samples. Ancylostoma caninum ova were used as surrogate for human hookworm (Ancylostoma duodenale and Necator americanus). Known concentration of A. caninum hookworm ova were seeded into wastewater (treated and raw) and sludge samples collected from two wastewater treatment plants (WWTPs) in Brisbane and Perth, Australia. The A. caninum ova were concentrated from treated and raw wastewater samples using centrifugation (Method A), hollow fiber ultrafiltration (HFUF) (Method B), filtration (Method C) and flotation (Method D) methods. For sludge samples, flotation (Method E) and direct DNA extraction (Method F) methods were used. Among the four methods tested, filtration (Method C) method was able to recover higher concentrations of A. caninum ova consistently from treated wastewater (39-50%) and raw wastewater (7.1-12%) samples collected from both WWTPs. The remaining methods (Methods A, B and D) yielded variable recovery rate ranging from 0.2 to 40% for treated and raw wastewater samples. The recovery rates for sludge samples were poor (0.02-4.7), although, Method F (direct DNA extraction) provided 1-2 orders of magnitude higher recovery rate than Method E (flotation). Based on our results it can be concluded that the recovery rates of hookworm ova from wastewater matrices, especially sludge samples, can be poor and highly variable. Therefore, choice of concentration method is vital for the sensitive detection of hookworm ova in wastewater matrices. Crown Copyright © 2015. Published by Elsevier

  10. Rapid Determination of Isomeric Benzoylpaeoniflorin and Benzoylalbiflorin in Rat Plasma by LC-MS/MS Method

    Directory of Open Access Journals (Sweden)

    Chuanqi Zhou

    2017-01-01

    Full Text Available Benzoylpaeoniflorin (BP is a potential therapeutic agent against oxidative stress related Alzheimer’s disease. In this study, a more rapid, selective, and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS method was developed to determine BP in rat plasma distinguishing with a monoterpene isomer, benzoylalbiflorin (BA. The method showed a linear response from 1 to 1000 ng/mL (r>0.9950. The precision of the interday and intraday ranged from 2.03 to 12.48% and the accuracy values ranged from −8.00 to 10.33%. Each running of the method could be finished in 4 minutes. The LC-MS/MS method was validated for specificity, linearity, precision, accuracy, recovery, and stability and was found to be acceptable for bioanalytical application. Finally, this fully validated method was successfully applied to a pharmacokinetic study in rats following oral administration.

  11. Multifrequency excitation method for rapid and accurate dynamic test of micromachined gyroscope chips.

    Science.gov (United States)

    Deng, Yan; Zhou, Bin; Xing, Chao; Zhang, Rong

    2014-10-17

    A novel multifrequency excitation (MFE) method is proposed to realize rapid and accurate dynamic testing of micromachined gyroscope chips. Compared with the traditional sweep-frequency excitation (SFE) method, the computational time for testing one chip under four modes at a 1-Hz frequency resolution and 600-Hz bandwidth was dramatically reduced from 10 min to 6 s. A multifrequency signal with an equal amplitude and initial linear-phase-difference distribution was generated to ensure test repeatability and accuracy. The current test system based on LabVIEW using the SFE method was modified to use the MFE method without any hardware changes. The experimental results verified that the MFE method can be an ideal solution for large-scale dynamic testing of gyroscope chips and gyroscopes.

  12. Multifrequency Excitation Method for Rapid and Accurate Dynamic Test of Micromachined Gyroscope Chips

    Directory of Open Access Journals (Sweden)

    Yan Deng

    2014-10-01

    Full Text Available A novel multifrequency excitation (MFE method is proposed to realize rapid and accurate dynamic testing of micromachined gyroscope chips. Compared with the traditional sweep-frequency excitation (SFE method, the computational time for testing one chip under four modes at a 1-Hz frequency resolution and 600-Hz bandwidth was dramatically reduced from 10 min to 6 s. A multifrequency signal with an equal amplitude and initial linear-phase-difference distribution was generated to ensure test repeatability and accuracy. The current test system based on LabVIEW using the SFE method was modified to use the MFE method without any hardware changes. The experimental results verified that the MFE method can be an ideal solution for large-scale dynamic testing of gyroscope chips and gyroscopes.

  13. A Microfluidic Channel Method for Rapid Drug-Susceptibility Testing of Pseudomonas aeruginosa

    Science.gov (United States)

    Matsumoto, Yoshimi; Grushnikov, Andrey; Kikuchi, Kazuma; Noji, Hiroyuki; Yamaguchi, Akihito; Yagi, Yasushi

    2016-01-01

    The recent global increase in the prevalence of antibiotic-resistant bacteria and lack of development of new therapeutic agents emphasize the importance of selecting appropriate antimicrobials for the treatment of infections. However, to date, the development of completely accelerated drug susceptibility testing methods has not been achieved despite the availability of a rapid identification method. We proposed an innovative rapid method for drug susceptibility testing for Pseudomonas aeruginosa that provides results within 3 h. The drug susceptibility testing microfluidic (DSTM) device was prepared using soft lithography. It consisted of five sets of four microfluidic channels sharing one inlet slot, and the four channels are gathered in a small area, permitting simultaneous microscopic observation. Antimicrobials were pre-introduced into each channel and dried before use. Bacterial suspensions in cation-adjusted Mueller–Hinton broth were introduced from the inlet slot and incubated for 3 h. Susceptibilities were microscopically evaluated on the basis of differences in cell numbers and shapes between drug-treated and control cells, using dedicated software. The results of 101 clinically isolated strains of P. aeruginosa obtained using the DSTM method strongly correlated with results obtained using the ordinary microbroth dilution method. Ciprofloxacin, meropenem, ceftazidime, and piperacillin caused elongation in susceptible cells, while meropenem also induced spheroplast and bulge formation. Morphological observation could alternatively be used to determine the susceptibility of P. aeruginosa to these drugs, although amikacin had little effect on cell shape. The rapid determination of bacterial drug susceptibility using the DSTM method could also be applicable to other pathogenic species, and it could easily be introduced into clinical laboratories without the need for expensive instrumentation. PMID:26872134

  14. A Microfluidic Channel Method for Rapid Drug-Susceptibility Testing of Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    Yoshimi Matsumoto

    Full Text Available The recent global increase in the prevalence of antibiotic-resistant bacteria and lack of development of new therapeutic agents emphasize the importance of selecting appropriate antimicrobials for the treatment of infections. However, to date, the development of completely accelerated drug susceptibility testing methods has not been achieved despite the availability of a rapid identification method. We proposed an innovative rapid method for drug susceptibility testing for Pseudomonas aeruginosa that provides results within 3 h. The drug susceptibility testing microfluidic (DSTM device was prepared using soft lithography. It consisted of five sets of four microfluidic channels sharing one inlet slot, and the four channels are gathered in a small area, permitting simultaneous microscopic observation. Antimicrobials were pre-introduced into each channel and dried before use. Bacterial suspensions in cation-adjusted Mueller-Hinton broth were introduced from the inlet slot and incubated for 3 h. Susceptibilities were microscopically evaluated on the basis of differences in cell numbers and shapes between drug-treated and control cells, using dedicated software. The results of 101 clinically isolated strains of P. aeruginosa obtained using the DSTM method strongly correlated with results obtained using the ordinary microbroth dilution method. Ciprofloxacin, meropenem, ceftazidime, and piperacillin caused elongation in susceptible cells, while meropenem also induced spheroplast and bulge formation. Morphological observation could alternatively be used to determine the susceptibility of P. aeruginosa to these drugs, although amikacin had little effect on cell shape. The rapid determination of bacterial drug susceptibility using the DSTM method could also be applicable to other pathogenic species, and it could easily be introduced into clinical laboratories without the need for expensive instrumentation.

  15. [Rapid determination of lactose, sucrose, glucose and fructose in foods by capillary zone electrophoresis with indirect ultraviolet detection].

    Science.gov (United States)

    Zhang, Huanhuan; Li, Jiang; Zhao, Shan; Ding, Xiaojing; Wang, Zhi

    2015-08-01

    A new and rapid method for the simultaneous determination of lactose, sucrose, glucose and fructose by capillary zone electrophoresis ( CZE) with indirect ultraviolet detection was developed. The separation was completed with an uncoated fused-silica capillary with 30.2 cm of total length (effective length of 20 cm) x 50 µm. The separation buffer consisted of 4 mmol/L potassium sorbate, 10 mmol/L sodium phosphate, 30 mmol/L NaOH (pH 12. 56) and 0. 5 mmol/L hexadecytrimethylammonium bromide (CTAB). The separation was performed at a voltage of -8 kV with the ultraviolet detection at 254 nm. The analysis of the four carbohydrates was completed within 10 min. The limits of detection (S/N= 3) for lactose, sucrose, glucose and fructose were 50, 75, 25 and 25 mg/L, and the limits of quantification (S/N = 10) were 150, 225, 75 and 75 mg/L, respectively. The average recoveries for the four carbohydrates were in the range of 87.0%-107.0% with the relative standard deviations of 1.2%-4.7%. No organic solvent was consumed throughout the whole process of the analysis. The method was used for the analysis of nine food samples and a quality control sample. The results demonstrated that the method is simple, rapid, accurate, and suitable for the routine analysis of the four carbohydrates in food samples.

  16. Adjustment of a rapid method for quantification of Fusarium spp. spore suspensions in plant pathology.

    Science.gov (United States)

    Caligiore-Gei, Pablo F; Valdez, Jorge G

    2015-01-01

    The use of a Neubauer chamber is a broadly employed method when cell suspensions need to be quantified. However, this technique may take a long time and needs trained personnel. Spectrophotometry has proved to be a rapid, simple and accurate method to estimate the concentration of spore suspensions of isolates of the genus Fusarium. In this work we present a linear formula to relate absorbance measurements at 530nm with the number of microconidia/ml in a suspension. Copyright © 2014 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  17. Rapid method for protein quantitation by Bradford assay after elimination of the interference of polysorbate 80.

    Science.gov (United States)

    Cheng, Yongfeng; Wei, Haiming; Sun, Rui; Tian, Zhigang; Zheng, Xiaodong

    2016-02-01

    Bradford assay is one of the most common methods for measuring protein concentrations. However, some pharmaceutical excipients, such as detergents, interfere with Bradford assay even at low concentrations. Protein precipitation can be used to overcome sample incompatibility with protein quantitation. But the rate of protein recovery caused by acetone precipitation is only about 70%. In this study, we found that sucrose not only could increase the rate of protein recovery after 1 h acetone precipitation, but also did not interfere with Bradford assay. So we developed a method for rapid protein quantitation in protein drugs even if they contained interfering substances. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Comparison of the NIDS® rapid assay with ELISA methods in immunogenicity testing of two biotherapeutics.

    Science.gov (United States)

    Pan, Jing; Small, Thomas; Qin, Dujie; Li, Shawn; Wang, Li; Chen, Dave; Pauley, Cindy; Verch, Thorsten; Kaplanski, Catherine; Bakhtiar, Ray; Vallejo, Yli Remo; Yin, Ray

    2011-01-01

    Rapid lateral flow immunogenicity assays for the detection of anti-drug antibodies (ADAs) to two biotherapeutic antibodies, an anti-HER2 antibody and an anti-TNF-α antibody, were developed using ANP Technologies, Inc.'s proprietary Nano-Intelligent Detection System (NIDS®) and compared to their ELISA counterparts. Biotin and hapten-labeled drugs are incubated with the patient serum sample to allow ADA to form a bridge complex with each drug conjugate. The reaction mixture is then added to a test strip with an anti-hapten capture zone which captures the mixed bridge complex. The bridge-complexed biotinylated drug then reacts with streptavidin-labeled gold particles in situ. The signal developed at the capture zone, which is directly proportional to ADA in the sample, is then quantitatively measured with a handheld reader. The counterpart ELISAs were run using the same reagents. Dose-response, specificity/free drug depletion, and screening cut-point assays were performed using both methods. The rapid assays' performance compare very closely to their ELISA counterparts'. Both types of assays identified the same positive samples in screening a limited population of 50 normal serum samples for the anti-HER2 antibody. In the case of anti-TNF-α, both assays identified the same positive samples out of 50 normal and 20 rheumatoid arthritis patient serum samples but differed in the assessment of two others. The rapid assay correctly identified as negative an ELISA false positive sample, and correctly tested as positive an ELISA false negative sample. Positive results were verified with a specificity/free drug depletion assay. The NIDS® rapid immunogenicity assay offers distinct advantages over current methods in simplicity, low cost, and short time to result. More importantly, the method requires no sample dilution and no washing steps which can perturb fragile complexes formed by low-affinity ADAs. Thus, the assay can potentially detect ADAs with various affinities

  19. A novel rapid and reproducible flow cytometric method for optimization of transfection efficiency in cells

    Science.gov (United States)

    Homann, Stefanie; Hofmann, Christian; Gorin, Aleksandr M.; Nguyen, Huy Cong Xuan; Huynh, Diana; Hamid, Phillip; Maithel, Neil; Yacoubian, Vahe; Mu, Wenli; Kossyvakis, Athanasios; Sen Roy, Shubhendu; Yang, Otto Orlean

    2017-01-01

    Transfection is one of the most frequently used techniques in molecular biology that is also applicable for gene therapy studies in humans. One of the biggest challenges to investigate the protein function and interaction in gene therapy studies is to have reliable monospecific detection reagents, particularly antibodies, for all human gene products. Thus, a reliable method that can optimize transfection efficiency based on not only expression of the target protein of interest but also the uptake of the nucleic acid plasmid, can be an important tool in molecular biology. Here, we present a simple, rapid and robust flow cytometric method that can be used as a tool to optimize transfection efficiency at the single cell level while overcoming limitations of prior established methods that quantify transfection efficiency. By using optimized ratios of transfection reagent and a nucleic acid (DNA or RNA) vector directly labeled with a fluorochrome, this method can be used as a tool to simultaneously quantify cellular toxicity of different transfection reagents, the amount of nucleic acid plasmid that cells have taken up during transfection as well as the amount of the encoded expressed protein. Finally, we demonstrate that this method is reproducible, can be standardized and can reliably and rapidly quantify transfection efficiency, reducing assay costs and increasing throughput while increasing data robustness. PMID:28863132

  20. A Rapid and Economical Method for Efficient DNA Extraction from Diverse Soils Suitable for Metagenomic Applications

    Science.gov (United States)

    Devi, Selvaraju Gayathri; Fathima, Anwar Aliya; Radha, Sudhakar; Arunraj, Rex; Curtis, Wayne R.; Ramya, Mohandass

    2015-01-01

    A rapid, cost effective method of metagenomic DNA extraction from soil is a useful tool for environmental microbiology. The present work describes an improved method of DNA extraction namely “powdered glass method” from diverse soils. The method involves the use of sterile glass powder for cell lysis followed by addition of 1% powdered activated charcoal (PAC) as purifying agent to remove humic substances. The method yielded substantial DNA (5.87 ± 0.04 μg/g of soil) with high purity (A260/280: 1.76 ± 0.05) and reduced humic substances (A340: 0.047 ± 0.03). The quality of the extracted DNA was compared against five different methods based on 16S rDNA PCR amplification, BamHI digestion and validated using quantitative PCR. The digested DNA was used for a metagenomic library construction with the transformation efficiency of 4 X 106 CFU mL-1. Besides providing rapid, efficient and economical extraction of metgenomic DNA from diverse soils, this method’s applicability is also demonstrated for cultivated organisms (Gram positive B. subtilis NRRL-B-201, Gram negative E. coli MTCC40, and a microalgae C. sorokiniana UTEX#1666). PMID:26167854

  1. New modelling method for fast reactor neutronic behaviours analysis; Nouvelles methodes de modelisation neutronique des reacteurs rapides de quatrieme Generation

    Energy Technology Data Exchange (ETDEWEB)

    Jacquet, P.

    2011-05-23

    Due to safety rules running on fourth generation reactors' core development, neutronics simulation tools have to be as accurate as never before. First part of this report enumerates every step of fast reactor's neutronics simulation implemented in current reference code: ECCO. Considering the field of fast reactors that meet criteria of fourth generation, ability of models to describe self-shielding phenomenon, to simulate neutrons leakage in a lattice of fuel assemblies and to produce representative macroscopic sections is evaluated. The second part of this thesis is dedicated to the simulation of fast reactors' core with steel reflector. These require the development of advanced methods of condensation and homogenization. Several methods are proposed and compared on a typical case: the ZONA2B core of MASURCA reactor. (author) [French] Les criteres de surete qui regissent le developpement de coeurs de reacteurs de quatrieme generation implique l'usage d'outils de calcul neutronique performants. Une premiere partie de la these reprend toutes les etapes de modelisation neutronique des reacteurs rapides actuellement d'usage dans le code de reference ECCO. La capacite des modeles a decrire le phenomene d'autoprotection, a representer les fuites neutroniques au niveau d'un reseau d'assemblages combustibles et a generer des sections macroscopiques representatives est appreciee sur le domaine des reacteurs rapides innovants respectant les criteres de quatrieme generation. La deuxieme partie de ce memoire se consacre a la modelisation des coeurs rapides avec reflecteur acier. Ces derniers necessitent le developpement de methodes avancees de condensation et d'homogenisation. Plusieurs methodes sont proposees et confrontees sur un probleme de modelisation typique: le coeur ZONA2B du reacteur maquette MASURCA

  2. A rapid Salmonella detection method involving thermophilic helicase-dependent amplification and a lateral flow assay.

    Science.gov (United States)

    Du, Xin-Jun; Zhou, Tian-Jiao; Li, Ping; Wang, Shuo

    2017-08-01

    Salmonella is a major foodborne pathogen that is widespread in the environment and can cause serious human and animal disease. Since conventional culture methods to detect Salmonella are time-consuming and laborious, rapid and accurate techniques to detect this pathogen are critically important for food safety and diagnosing foodborne illness. In this study, we developed a rapid, simple and portable Salmonella detection strategy that combines thermophilic helicase-dependent amplification (tHDA) with a lateral flow assay to provide a detection result based on visual signals within 90 min. Performance analyses indicated that the method had detection limits for DNA and pure cultured bacteria of 73.4-80.7 fg and 35-40 CFU, respectively. Specificity analyses showed no cross reactions with Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Enterobacter aerogenes, Shigella and Campylobacter jejuni. The results for detection in real food samples showed that 1.3-1.9 CFU/g or 1.3-1.9 CFU/mL of Salmonella in contaminated chicken products and infant nutritional cereal could be detected after 2 h of enrichment. The same amount of Salmonella in contaminated milk could be detected after 4 h of enrichment. This tHDA-strip can be used for the rapid detection of Salmonella in food samples and is particularly suitable for use in areas with limited equipment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Rapid methods to detect organic mercury and total selenium in biological samples

    Directory of Open Access Journals (Sweden)

    Basu Niladri

    2011-01-01

    Full Text Available Abstract Background Organic mercury (Hg is a global pollutant of concern and selenium is believed to afford protection against mercury risk though few approaches exist to rapidly assess both chemicals in biological samples. Here, micro-scale and rapid methods to detect organic mercury ( Results For organic Hg, samples are digested using Tris-HCl buffer (with sequential additions of protease, NaOH, cysteine, CuSO4, acidic NaBr followed by extraction with toluene and Na2S2O3. The final product is analyzed via commercially available direct/total mercury analyzers. For Se, a fluorometric assay has been developed for microplate readers that involves digestion (HNO3-HClO4 and HCl, conjugation (2,3-diaminonaphthalene, and cyclohexane extraction. Recovery of organic Hg (86-107% and Se (85-121% were determined through use of Standard Reference Materials and lemon shark kidney tissues. Conclusions The approaches outlined provide an easy, rapid, reproducible, and cost-effective platform for monitoring organic Hg and total Se in biological samples. Owing to the importance of organic Hg and Se in the pathophysiology of Hg, integration of such methods into established research monitoring efforts (that largely focus on screening total Hg only will help increase understanding of Hg's true risks.

  4. Application of rapid cloud point extraction method for trace cobalt analysis coupled with spectrophotometric determination

    Science.gov (United States)

    Wen, Xiaodong; He, Lei; Shi, Chunsheng; Deng, Qingwen; Wang, Jiwei; Zhao, Xia

    2013-11-01

    In this work, the analytical performance of conventional spectrophotometer was improved through the coupling of effective preconcentration method with spectrophotometric determination. Rapidly synergistic cloud point extraction (RS-CPE) was used to pre-concentrate ultra trace cobalt and firstly coupled with spectrophotometric determination. The developed coupling was simple, rapid and efficient. The factors influencing RS-CPE and spectrophotometer were optimized. Under the optimal conditions, the limit of detection (LOD) was 0.6 μg L-1, with sensitivity enhancement factor of 23. The relative standard deviation (RSD) for seven replicate measurements of 50 μg L-1 of cobalt was 4.3%. The recoveries for the spiked samples were in the acceptable range of 93.8-105%.

  5. A rapid and direct real time PCR-based method for identification of Salmonella spp

    DEFF Research Database (Denmark)

    Rodriguez-Lazaro, D.; Hernández, Marta; Esteve, T.

    2003-01-01

    The aim of this work was the validation of a rapid, real-time PCR assay based on TaqMan((R)) technology for the unequivocal identification of Salmonella spp. to be used directly on an agar-grown colony. A real-time PCR system targeting at the Salmonella spp. invA gene was optimized and validated...... to be especially convenient because the pre-mix containing all PCR reagents except for the bacterial cells could be kept at -20 degreesC for at least I month before its use. The optimized TaqMan((R)) real-time PCR assay is a useful, simple and rapid method for routine identification of Salmonella spp...

  6. A rapid method for counting nucleated erythrocytes on stained blood smears by digital image analysis

    Science.gov (United States)

    Gering, E.; Atkinson, C.T.

    2004-01-01

    Measures of parasitemia by intraerythrocytic hematozoan parasites are normally expressed as the number of infected erythrocytes per n erythrocytes and are notoriously tedious and time consuming to measure. We describe a protocol for generating rapid counts of nucleated erythrocytes from digital micrographs of thin blood smears that can be used to estimate intensity of hematozoan infections in nonmammalian vertebrate hosts. This method takes advantage of the bold contrast and relatively uniform size and morphology of erythrocyte nuclei on Giemsa-stained blood smears and uses ImageJ, a java-based image analysis program developed at the U.S. National Institutes of Health and available on the internet, to recognize and count these nuclei. This technique makes feasible rapid and accurate counts of total erythrocytes in large numbers of microscope fields, which can be used in the calculation of peripheral parasitemias in low-intensity infections.

  7. [Three-Iindex-Value Method for Rapid Screening Unqualified Vegetable Oil].

    Science.gov (United States)

    He, Wen-xuan; Hong, Gui-shui; Fang, Run; Cai, Xian-chun; Huang, Sheng

    2015-04-01

    In the present study, by measuring the A3 005 (representing unsaturation), A985 (representing conjugated fatty acids), A960 + A985 (representing trans-fatty acid ) of southern common vegetable oils (peanut oil, corn oil, canola oil, soybean oil, sunflower oil, tea seed oil and olive oil), "waste oil" and overdue vegetable oils, the pass-setting-range of these three index values for the vegetable oils was obtained. On this basis, a method for rapid screening unqualified vegetable oil (expired, adding low-cost oil, adding "waste oil") was established. The method effectively improved the monitoring efficiency of vegetable oil. With this method of screening a number of suspected substandard oils were proved unqualified by determination of fatty acid composition and 11, 12, 13, 17 fatty acid content. Through the combination of several detection methods, the causes for disqualification of vegetable oils can be further inferred.

  8. New rapid DNA extraction method with Chelex from Venturia inaequalis spores.

    Science.gov (United States)

    Turan, Ceren; Nanni, Irene Maja; Brunelli, Agostino; Collina, Marina

    2015-08-01

    The objective of this study was to develop a rapid method to isolate DNA from Venturia inaequalis spores for use in diagnostic DNA mutation analysis. Chelex-100 resin was evaluated and compared with a well established DNA exctraction method, utilizing CTAB in order to have a robust comparison. In this research we demonstrated that Chelex-100 efficiently makes extraction of the DNA from V. inaequalis spores available for direct use in molecular analyses. Also, the quantity and quality of extracted DNA were shown to be adequate for PCR analysis. Comparatively, the quality of DNA samples isolated using Chelex method was better than those extracted using CTAB. In conclusion, the Chelex method is recommended for PCR experiments considering its simplicity and cost-effectiveness. Copyright © 2015. Published by Elsevier B.V.

  9. Rapid and sensitive liquid chromatography-mass spectrometry method for determination of ropinirole in human plasma.

    Science.gov (United States)

    Bhatt, Jignesh; Jangid, Arvind; Shetty, Raghavendra; Shah, Bhavin; Kambli, Sandeep; Subbaiah, Gunta; Singh, Sadhana

    2006-03-18

    A rapid and robust liquid chromatography-mass spectrometry (LC-MS/MS) method was developed for non-ergoline dopamine D(2)-receptor agonist, ropinirole in human plasma using Es-citalopram oxalate as an internal standard. The method involves solid phase extraction from plasma, reversed-phase simple isocratic chromatographic conditions and mass spectrometric detection that enables a detection limit at picogram levels. The proposed method was validated with linear range of 20-1,200 pg/ml. The extraction recoveries for ropinirole and internal standard were 90.45 and 65.42%, respectively. The R.S.D.% of intra-day and inter-day assay was lower than 15%. For its sensitivity and reliability, the proposed method is particularly suitable for pharmacokinetic studies.

  10. Simple and rapid methods for detecting Salmonella enteritidis in raw eggs.

    Science.gov (United States)

    Seo, Kun-Ho; Holt, Peter S; Stone, Henry D; Gast, Richard K

    2003-10-15

    The Centers for Disease Control and Prevention estimates there were 300,000 cases of Salmonella enteritidis (SE) in 1997. Egg products were associated with many of the cases. To address this problem, many producers implemented flock surveillance of the SE situation at their facilities. A rapid and simple method for detecting SE from poultry samples is critical for the effective implementation of such testing strategies. A lateral flow device for the detection of SE utilized in this study was manufactured by Neogen, Lansing, MI. The test panel is a presumptive qualitative test system that detects only members of Group D1 Salmonella species. A series of studies were conducted to optimize the test procedure for raw eggs with different sample preparations. A novel antigen extraction method was developed for use with the test panel kit. The detection limit of the test panel kit was increased approximately tenfold when the extraction method was used. Detection of SE was 100% in raw egg pools inoculated with 10 SE cells per ml of egg and incubated at a 1:10 ratio in buffered peptone water (BPW) or tetrathionate brilliant green broth (TBG) for 24 h at 37 degrees C. The developed lateral flow test kit could provide a simple, rapid, and inexpensive method for egg producers and processors to test specifically for Salmonella group D1 serovars, such as SE, in egg samples.

  11. Note: Non-invasive optical method for rapid determination of alignment degree of oriented nanofibrous layers

    Energy Technology Data Exchange (ETDEWEB)

    Pokorny, M.; Rebicek, J. [R& D Department, Contipro Biotech s.r.o., 561 02 Dolni Dobrouc (Czech Republic); Klemes, J. [R& D Department, Contipro Pharma a.s., 561 02 Dolni Dobrouc (Czech Republic); Kotzianova, A. [R& D Department, Contipro Pharma a.s., 561 02 Dolni Dobrouc (Czech Republic); Department of Chemistry, Faculty of Science, Masaryk University, Kamenice 5, CZ-62500 Brno (Czech Republic); Velebny, V. [R& D Department, Contipro Biotech s.r.o., 561 02 Dolni Dobrouc (Czech Republic); R& D Department, Contipro Pharma a.s., 561 02 Dolni Dobrouc (Czech Republic)

    2015-10-15

    This paper presents a rapid non-destructive method that provides information on the anisotropic internal structure of nanofibrous layers. A laser beam of a wavelength of 632.8 nm is directed at and passes through a nanofibrous layer prepared by electrostatic spinning. Information about the structural arrangement of nanofibers in the layer is directly visible in the form of a diffraction image formed on a projection screen or obtained from measured intensities of the laser beam passing through the sample which are determined by the dependency of the angle of the main direction of polarization of the laser beam on the axis of alignment of nanofibers in the sample. Both optical methods were verified on Polyvinyl alcohol (PVA) nanofibrous layers (fiber diameter of 470 nm) with random, single-axis aligned and crossed structures. The obtained results match the results of commonly used methods which apply the analysis of electron microscope images. The presented simple method not only allows samples to be analysed much more rapidly and without damaging them but it also makes possible the analysis of much larger areas, up to several square millimetres, at the same time.

  12. Rapid intrinsic fluorescence method for direct identification of pathogens in blood cultures.

    Science.gov (United States)

    Walsh, John D; Hyman, Jay M; Borzhemskaya, Larisa; Bowen, Ann; McKellar, Caroline; Ullery, Michael; Mathias, Erin; Ronsick, Christopher; Link, John; Wilson, Mark; Clay, Bradford; Robinson, Ron; Thorpe, Thurman; van Belkum, Alex; Dunne, W Michael

    2013-11-19

    A positive blood culture is a critical result that requires prompt identification of the causative agent. This article describes a simple method to identify microorganisms from positive blood culture broth within the time taken to perform a Gram stain (identification of the etiologic agent may benefit the clinical management of sepsis. Further evaluation is now warranted to determine the performance of the method using clinical blood culture specimens. Physicians often require the identity of the infective agent in order to make life-saving adjustments to empirical therapy or to switch to less expensive and/or more targeted antimicrobials. However, standard identification procedures take up to 2 days after a blood culture is signaled positive, and even most rapid molecular techniques take several hours to provide a result. Other techniques are faster (e.g., matrix-assisted laser desorption ionization-time of flight [MALDI-TOF] mass spectrometry) but require time-consuming manual processing steps and expensive equipment. There remains a clear need for a simple, inexpensive method to rapidly identify microorganisms directly from positive blood cultures. The promising new method described in this research article can identify microorganisms in minutes by optical spectroscopy, thus permitting the lab to simultaneously report the presence of a positive blood culture and the organism's identity.

  13. Use of refractometry and colorimetry as field methods to rapidly assess antimalarial drug quality.

    Science.gov (United States)

    Green, Michael D; Nettey, Henry; Villalva Rojas, Ofelia; Pamanivong, Chansapha; Khounsaknalath, Lamphet; Grande Ortiz, Miguel; Newton, Paul N; Fernández, Facundo M; Vongsack, Latsamy; Manolin, Ot

    2007-01-04

    The proliferation of counterfeit and poor-quality drugs is a major public health problem; especially in developing countries lacking adequate resources to effectively monitor their prevalence. Simple and affordable field methods provide a practical means of rapidly monitoring drug quality in circumstances where more advanced techniques are not available. Therefore, we have evaluated refractometry, colorimetry and a technique combining both processes as simple and accurate field assays to rapidly test the quality of the commonly available antimalarial drugs; artesunate, chloroquine, quinine, and sulfadoxine. Method bias, sensitivity, specificity and accuracy relative to high-performance liquid chromatographic (HPLC) analysis of drugs collected in the Lao PDR were assessed for each technique. The HPLC method for each drug was evaluated in terms of assay variability and accuracy. The accuracy of the combined method ranged from 0.96 to 1.00 for artesunate tablets, chloroquine injectables, quinine capsules, and sulfadoxine tablets while the accuracy was 0.78 for enterically coated chloroquine tablets. These techniques provide a generally accurate, yet simple and affordable means to assess drug quality in resource-poor settings.

  14. A two-step method for rapid characterization of electroosmotic flows in capillary electrophoresis.

    Science.gov (United States)

    Zhang, Wenjing; He, Muyi; Yuan, Tao; Xu, Wei

    2017-12-01

    The measurement of electroosmotic flow (EOF) is important in a capillary electrophoresis (CE) experiment in terms of performance optimization and stability improvement. Although several methods exist, there are demanding needs to accurately characterize ultra-low electroosmotic flow rates (EOF rates), such as in coated capillaries used in protein separations. In this work, a new method, called the two-step method, was developed to accurately and rapidly measure EOF rates in a capillary, especially for measuring the ultra-low EOF rates in coated capillaries. In this two-step method, the EOF rates were calculated by measuring the migration time difference of a neutral marker in two consecutive experiments, in which a pressure driven was introduced to accelerate the migration and the DC voltage was reversed to switch the EOF direction. Uncoated capillaries were first characterized by both this two-step method and a conventional method to confirm the validity of this new method. Then this new method was applied in the study of coated capillaries. Results show that this new method is not only fast in speed, but also better in accuracy. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Non-supervised method for early forest fire detection and rapid mapping

    Science.gov (United States)

    Artés, Tomás; Boca, Roberto; Liberta, Giorgio; San-Miguel, Jesús

    2017-09-01

    Natural hazards are a challenge for the society. Scientific community efforts have been severely increased assessing tasks about prevention and damage mitigation. The most important points to minimize natural hazard damages are monitoring and prevention. This work focuses particularly on forest fires. This phenomenon depends on small-scale factors and fire behavior is strongly related to the local weather. Forest fire spread forecast is a complex task because of the scale of the phenomena, the input data uncertainty and time constraints in forest fire monitoring. Forest fire simulators have been improved, including some calibration techniques avoiding data uncertainty and taking into account complex factors as the atmosphere. Such techniques increase dramatically the computational cost in a context where the available time to provide a forecast is a hard constraint. Furthermore, an early mapping of the fire becomes crucial to assess it. In this work, a non-supervised method for forest fire early detection and mapping is proposed. As main sources, the method uses daily thermal anomalies from MODIS and VIIRS combined with land cover map to identify and monitor forest fires with very few resources. This method relies on a clustering technique (DBSCAN algorithm) and on filtering thermal anomalies to detect the forest fires. In addition, a concave hull (alpha shape algorithm) is applied to obtain rapid mapping of the fire area (very coarse accuracy mapping). Therefore, the method leads to a potential use for high-resolution forest fire rapid mapping based on satellite imagery using the extent of each early fire detection. It shows the way to an automatic rapid mapping of the fire at high resolution processing as few data as possible.

  16. Interaction between Rivers and Aquifers: a method for rapid Identification of Transience in Streambed Conductance

    Science.gov (United States)

    Gianni, Guillaume; Perrochet, Pierre; Vogel, Alexandre; Brunner, Philip

    2016-04-01

    Streambed hydraulic conductance controls the interactions between surface water and groundwater. In order to quantify river-aquifer dynamics, quantifying conductance is indispensable. However, the streambed conductance is often subject to transience, as a result of the erosion and deposition processes in rivers. This transience has to be quantified and considered for any approach (i.e. numerical or analytical models) aimed at quantifying exchange fluxes. Directly measuring hydraulic properties in a river yields only point values, is time-consuming and therefore not suited to detect transience of the physical properties. We present a method to continuously and rapidly monitor transience of streambed conductance. Input data are time series of stream stage and hydraulic head variations in the aquifer. The method is based on the inversion of a floodwave response. The analytical model consists of only 3 parameters: x, the distance between streambank and an observation well, α, the aquifer diffusivity and a, the retardation coefficient that is inversely proportional to the streambed conductance. Estimation of a is carried out over successive time steps in order to identify transience in streambed conductance. The method is tested on synthetic data and is applied to field data from the Rhône River and its alluvial aquifer (Switzerland). The synthetic method demonstrated the robustness of the proposed methodology. Application of the method to the field site allowed identifying transience in streambed properties, following flood events in the Rhône. This method requires transience in the surface water, and the river should not change its width significantly with a rising water level. If these conditions are fulfilled, this method allows for a rapid and effective identification of transience of streambed conductance.

  17. The BUME method: a new rapid and simple chloroform-free method for total lipid extraction of animal tissue

    Science.gov (United States)

    Löfgren, Lars; Forsberg, Gun-Britt; Ståhlman, Marcus

    2016-06-01

    In this study we present a simple and rapid method for tissue lipid extraction. Snap-frozen tissue (15-150 mg) is collected in 2 ml homogenization tubes. 500 μl BUME mixture (butanol:methanol [3:1]) is added and automated homogenization of up to 24 frozen samples at a time in less than 60 seconds is performed, followed by a 5-minute single-phase extraction. After the addition of 500 μl heptane:ethyl acetate (3:1) and 500 μl 1% acetic acid a 5-minute two-phase extraction is performed. Lipids are recovered from the upper phase by automated liquid handling using a standard 96-tip robot. A second two-phase extraction is performed using 500 μl heptane:ethyl acetate (3:1). Validation of the method showed that the extraction recoveries for the investigated lipids, which included sterols, glycerolipids, glycerophospholipids and sphingolipids were similar or better than for the Folch method. We also applied the method for lipid extraction of liver and heart and compared the lipid species profiles with profiles generated after Folch and MTBE extraction. We conclude that the BUME method is superior to the Folch method in terms of simplicity, through-put, automation, solvent consumption, economy, health and environment yet delivering lipid recoveries fully comparable to or better than the Folch method.

  18. Rapid detection of carbapenemase production in Enterobacteriaceae using a modified paper strip Carba NP method.

    Science.gov (United States)

    Ho, Pak-Leung; Wang, Ya; Tse, Cindy Wing-Sze; Fung, Kitty Sau-Chun; Cheng, Vincent Ch-Chung; Lee, Rodney; To, Wing-Kin; Lai, Raymond Wai-Man; Luk, Wei-Kwang; Que, Tak-Lun; Tsang, Dominic Ngai-Chong

    2017-10-25

    Rapid and accurate detection of carbapenemase-producing Enterobacteriaceae (CPE) is important for preventing their spread in healthcare settings. We compared the performance of the Carba NP test using the CLSI tube method with that using a modified paper strip method for detection of carbapenemase in 390 Enterobacteriaceae isolates. The isolates were identified by Hong Kong's carbapenem-resistant Enterobacteriaceae surveillance program in 2016 and comprised 213 CPE and 177 carbapenemase-negative Enterobacteriaceae Molecular genotype was used as the reference. Test results were read at different time points for the CLSI method (1 min, 5 min, 1 h and 2 h) and strip method (1 min, 5 min). The strip CNP and CLSI CNP tests correctly detect carbapenemase production in 93% and 93% KPC producers, 100% and 38% IMI producers, 94% and 85% IMP producers, 98% and 90% NDM producers, and, 29% and 12% OXA producers, respectively. Overall, the strip method has superior sensitivity than the CLSI method (86% vs. 75%, respectively, P NP test using the modified strip method has a higher sensitivity and a shorter assay time than using the CLSI tube method. Copyright © 2017 American Society for Microbiology.

  19. Rapid Staining Method to Detect and Identify Downy Mildew (Peronospora belbahrii in Basil

    Directory of Open Access Journals (Sweden)

    Adolfina R. Koroch

    2013-07-01

    Full Text Available Premise of the study: Demand for fresh-market sweet basil continues to increase, but in 2009 a new pathogen emerged, threatening commercial field/greenhouse production and leading to high crop losses. This study describes a simple and effective staining method for rapid microscopic detection of basil downy mildew (Peronospora belbahrii from leaves of basil (Ocimum basilicum. Methods and Results: Fresh leaf sections infected with P. belbahrii were placed on a microscope slide, cleared with Visikol™, and stained with iodine solution followed by one drop of 70% sulfuric acid. Cell walls of the pathogen were stained with a distinct coloration, providing a high-contrast image between the pathogen and plant. Conclusions: This new staining method can be used successfully to identify downy mildew in basil, which then can significantly reduce its spread if identified early, coupled with mitigation strategies. This technique can facilitate the control of the disease, without expensive and specialized equipment.

  20. The method of parallel-hierarchical transformation for rapid recognition of dynamic images using GPGPU technology

    Science.gov (United States)

    Timchenko, Leonid; Yarovyi, Andrii; Kokriatskaya, Nataliya; Nakonechna, Svitlana; Abramenko, Ludmila; Ławicki, Tomasz; Popiel, Piotr; Yesmakhanova, Laura

    2016-09-01

    The paper presents a method of parallel-hierarchical transformations for rapid recognition of dynamic images using GPU technology. Direct parallel-hierarchical transformations based on cluster CPU-and GPU-oriented hardware platform. Mathematic models of training of the parallel hierarchical (PH) network for the transformation are developed, as well as a training method of the PH network for recognition of dynamic images. This research is most topical for problems on organizing high-performance computations of super large arrays of information designed to implement multi-stage sensing and processing as well as compaction and recognition of data in the informational structures and computer devices. This method has such advantages as high performance through the use of recent advances in parallelization, possibility to work with images of ultra dimension, ease of scaling in case of changing the number of nodes in the cluster, auto scan of local network to detect compute nodes.

  1. Large Rapidity Gap Method to Select Soft Diffraction Dissociation at the LHC

    Directory of Open Access Journals (Sweden)

    Emily Nurse

    2016-01-01

    Full Text Available In proton-proton (pp collisions, any process involves exchanging the vacuum quantum numbers is known as diffractive process. A diffractive process with no large Q2 is called soft diffractive process. The diffractive processes are important for understanding nonperturbative QCD effects and they also constitute a significant fraction of the total pp cross section. The diffractive events are typically characterized by a region of the detector without particles, known as a rapidity gap. In order to observe diffractive events in this way, we consider the pseudorapidity acceptance in the forward region of the ATLAS and CMS detectors at the Large Hadron Collider (LHC and discuss the methods to select soft diffractive dissociation for pp collisions at s=7 TeV. It is shown that, in the limited detector rapidity acceptance, it is possible to select diffractive dissociation events by requiring a rapidity gap in the event; however, without using forward detectors, it seems not possible to fully separate single and double diffractive dissociation events. The Zero Degree Calorimeters can be used to distinguish the type of the diffractive processes up to a certain extent.

  2. Hierarchically rough, mechanically durable and superhydrophobic epoxy coatings through rapid evaporation spray method

    Energy Technology Data Exchange (ETDEWEB)

    Simovich, Tomer; Wu, Alex H.; Lamb, Robert N., E-mail: rnlamb@unimelb.edu.au

    2015-08-31

    A mechanically durable and scalable superhydrophobic coating was fabricated by combining the advantages of both bottom-up and top-down approaches into a one-pot, one-step application method. This is achieved by spray coating a solution consisting of silica nanoparticles, which are embedded within epoxy resin, onto a heated substrate to rapidly drive both solvent evaporation and curing simultaneously. By maintaining a high substrate temperature, the arrival of spray-delivered micrometer-sized droplets are rapidly cured onto the substrate to form surface microroughness, while simultaneously, rapid solvent evaporation within each droplet results in the formation of a nanoporous structure. SEM, dual-beam FIB, and cross-sectional TEM/EDAX elemental mapping were used to confirm both the chemistry and the requisite micro- and nano-porosity within the coating structure requisite for superhydrophobicity. The resultant coatings exhibit contact angles greater than 150° (153.8° ± 0.8°) and roll-off angles of 8° ± 2°, with a coating hardness of 6H on the pencil hardness scale, and a rating of 5 on an ASTM crosshatch test. - Highlights: • A highly superhydrophobic coating was fabricated utilizing epoxy and nanoparticles. • The coating was demonstrated to be very durable and abrasion resistant. • The fabrication involves a novel, scalable one-pot synthesis technique.

  3. Rapid Method for Ra-226 and Ra-228 in Water Samples

    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, Sherrod, L. III

    2006-02-10

    The measurement of radium isotopes in natural waters is important for oceanographic studies and for public health reasons. Ra-226 (1620 year half-life) is one of the most toxic of the long-lived alpha emitters present in the environment due to its long life and its tendency to concentrate in bones, which increases the internal radiation dose of individuals. The analysis of radium-226 and radium-228 in natural waters can be tedious and time-consuming. Different sample preparation methods are often required to prepare Ra-226 and Ra-228 for separate analyses. A rapid method has been developed at the Savannah River Environmental Laboratory that effectively separates both Ra-226 and Ra-228 (via Ac-228) for assay. This method uses MnO{sub 2} Resin from Eichrom Technologies (Darien, IL, USA) to preconcentrate Ra-226 and Ra-228 rapidly from water samples, along with Ba-133 tracer. DGA Resin{reg_sign} (Eichrom) and Ln-Resin{reg_sign} (Eichrom) are employed in tandem to prepare Ra-226 for assay by alpha spectrometry and to determine Ra-228 via the measurement of Ac-228 by gas proportional counting. After preconcentration, the manganese dioxide is dissolved from the resin and passed through stacked Ln-Resin-DGA Resin cartridges that remove uranium and thorium interferences and retain Ac-228 on DGA Resin. The eluate that passed through this column is evaporated, redissolved in a lower acidity and passed through Ln-Resin again to further remove interferences before performing a barium sulfate microprecipitation. The Ac-228 is stripped from the resin, collected using cerium fluoride microprecipitation and counted by gas proportional counting. By using vacuum box cartridge technology with rapid flow rates, sample preparation time is minimized.

  4. A rapid and sensitive method for measuring N-acetylglucosaminidase activity in cultured cells.

    Directory of Open Access Journals (Sweden)

    Victor Mauri

    Full Text Available A rapid and sensitive method to quantitatively assess N-acetylglucosaminidase (NAG activity in cultured cells is highly desirable for both basic research and clinical studies. NAG activity is deficient in cells from patients with Mucopolysaccharidosis type IIIB (MPS IIIB due to mutations in NAGLU, the gene that encodes NAG. Currently available techniques for measuring NAG activity in patient-derived cell lines include chromogenic and fluorogenic assays and provide a biochemical method for the diagnosis of MPS IIIB. However, standard protocols require large amounts of cells, cell disruption by sonication or freeze-thawing, and normalization to the cellular protein content, resulting in an error-prone procedure that is material- and time-consuming and that produces highly variable results. Here we report a new procedure for measuring NAG activity in cultured cells. This procedure is based on the use of the fluorogenic NAG substrate, 4-Methylumbelliferyl-2-acetamido-2-deoxy-alpha-D-glucopyranoside (MUG, in a one-step cell assay that does not require cell disruption or post-assay normalization and that employs a low number of cells in 96-well plate format. We show that the NAG one-step cell assay greatly discriminates between wild-type and MPS IIIB patient-derived fibroblasts, thus providing a rapid method for the detection of deficiencies in NAG activity. We also show that the assay is sensitive to changes in NAG activity due to increases in NAGLU expression achieved by either overexpressing the transcription factor EB (TFEB, a master regulator of lysosomal function, or by inducing TFEB activation chemically. Because of its small format, rapidity, sensitivity and reproducibility, the NAG one-step cell assay is suitable for multiple procedures, including the high-throughput screening of chemical libraries to identify modulators of NAG expression, folding and activity, and the investigation of candidate molecules and constructs for applications in

  5. Rapid expansion method (REM) for time‐stepping in reverse time migration (RTM)

    KAUST Repository

    Pestana, Reynam C.

    2009-01-01

    We show that the wave equation solution using a conventional finite‐difference scheme, derived commonly by the Taylor series approach, can be derived directly from the rapid expansion method (REM). After some mathematical manipulation we consider an analytical approximation for the Bessel function where we assume that the time step is sufficiently small. From this derivation we find that if we consider only the first two Chebyshev polynomials terms in the rapid expansion method we can obtain the second order time finite‐difference scheme that is frequently used in more conventional finite‐difference implementations. We then show that if we use more terms from the REM we can obtain a more accurate time integration of the wave field. Consequently, we have demonstrated that the REM is more accurate than the usual finite‐difference schemes and it provides a wave equation solution which allows us to march in large time steps without numerical dispersion and is numerically stable. We illustrate the method with post and pre stack migration results.

  6. An evaluation of rapid methods for monitoring vegetation characteristics of wetland bird habitat

    Science.gov (United States)

    Tavernia, Brian G.; Lyons, James E.; Loges, Brian W.; Wilson, Andrew; Collazo, Jaime A.; Runge, Michael C.

    2016-01-01

    Wetland managers benefit from monitoring data of sufficient precision and accuracy to assess wildlife habitat conditions and to evaluate and learn from past management decisions. For large-scale monitoring programs focused on waterbirds (waterfowl, wading birds, secretive marsh birds, and shorebirds), precision and accuracy of habitat measurements must be balanced with fiscal and logistic constraints. We evaluated a set of protocols for rapid, visual estimates of key waterbird habitat characteristics made from the wetland perimeter against estimates from (1) plots sampled within wetlands, and (2) cover maps made from aerial photographs. Estimated percent cover of annuals and perennials using a perimeter-based protocol fell within 10 percent of plot-based estimates, and percent cover estimates for seven vegetation height classes were within 20 % of plot-based estimates. Perimeter-based estimates of total emergent vegetation cover did not differ significantly from cover map estimates. Post-hoc analyses revealed evidence for observer effects in estimates of annual and perennial covers and vegetation height. Median time required to complete perimeter-based methods was less than 7 percent of the time needed for intensive plot-based methods. Our results show that rapid, perimeter-based assessments, which increase sample size and efficiency, provide vegetation estimates comparable to more intensive methods.

  7. wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

    Science.gov (United States)

    Passet, Virginie; Haugaard, Anita Björk; Babosan, Anamaria; Kassis-Chikhani, Najiby; Struve, Carsten; Decré, Dominique

    2013-01-01

    Pathogens of the genus Klebsiella have been classified into distinct capsular (K) types for nearly a century. K typing of Klebsiella species still has important applications in epidemiology and clinical microbiology, but the serological method has strong practical limitations. Our objective was to evaluate the sequencing of wzi, a gene conserved in all capsular types of Klebsiella pneumoniae that codes for an outer membrane protein involved in capsule attachment to the cell surface, as a simple and rapid method for the prediction of K type. The sequencing of a 447-nucleotide region of wzi distinguished the K-type reference strains with only nine exceptions. A reference wzi sequence database was created by the inclusion of multiple strains representing K types associated with high virulence and multidrug resistance. A collection of 119 prospective clinical isolates of K. pneumoniae were then analyzed in parallel by wzi sequencing and classical K typing. Whereas K typing achieved typeability for 81% and discrimination for 94.4% of the isolates, these figures were 98.1% and 98.3%, respectively, for wzi sequencing. The prediction of K type once the wzi allele was known was 94%. wzi sequencing is a rapid and simple method for the determination of the K types of most K. pneumoniae clinical isolates. PMID:24088853

  8. A rapid method for measuring soil water content in the field with a areometer

    Directory of Open Access Journals (Sweden)

    Calbo Adonai Gimenez

    2002-01-01

    Full Text Available The availability of a rapid method to evaluate the soil water content (U can be an important tool to determine the moment to irrigate. The soil areometer consists of an elongated hydrostatic balance with a weighing pan, a graduated neck, a float and a pynometric flask. In this work an areometer was adapted to rapidly measure soil water content without the need of drying the soil. The expression U = (M A - M AD/(M M -M A was used to calculate the soil water content. In this equation M M is the mass to level the areometer with the pycnometric flask filled with water, M A the mass to level the areometer with a mass M M of soil in the pycnometer, the volume being completed with water, and similarly M AD the mass added to the pan to level the areometer with a mass M M of dried soil in the pycnometric flask. The convenience of this method is that the values M M and M AD are known. Consequently, the decision on irrigation can be made after a measurement that takes, about, ten minutes. The procedure involves only stirring the soil with water for at least 2 minutes to remove the adhered air. The soil water content data obtained with the areometric method were similar to those obtained weighing the soil before and after drying to constant weight, in an oven at 105º C.

  9. Single-step blood direct PCR: A robust and rapid method to diagnose triplet repeat disorders.

    Science.gov (United States)

    Singh, Inder; Swarup, Vishnu; Shakya, Sunil; Goyal, Vinay; Faruq, Mohammed; Srivastava, Achal Kumar

    2017-08-15

    DNA extraction prior to polymerase chain reaction (PCR) amplification in genetic diagnoses of triplet repeat disorders (TRDs) is tedious and labour-intensive and has the limitations of sample contamination with foreign DNA, including that from preceding samples. Therefore, we aimed to develop a rapid, robust, and cost-effective method for expeditious genetic investigation of TRDs from whole blood as a DNA template. Peripheral blood samples were collected from 70 clinically suspected patients of progressive ataxia. The conventional method using genomic DNA and single-step Blood-Direct PCR (BD-PCR) method with just 2μl of whole blood sample were tested to amplify triplet repeat expansion in genes related to spinocerebellar ataxia (SCA) types 1, 2, 3, 12 and Friedreich's ataxia (FRDA). Post-PCR, the allele sizes were mapped and repeat numbers were calculated using GeneMapper and macros run in Microsoft Excel programmes. Successful amplification of target regions was achieved in all samples by both methods. The frequency of the normal and mutated allele was concordant between both methods, diagnosing 37% positive for a mutation in either of the candidate genes. The BD-PCR resulted in higher intensities of product peaks of normal and pathogenic alleles. The nearly-accurate sizing of the normal and expanded allele was achieved in a shorter time (4-5h), without DNA extraction and any risk of cross contamination, which suggests the BD-PCR to be a reliable, inexpensive, and rapid method to confirm TRDs. This technique can be introduced in routine diagnostic procedures of other tandem repeat disorders. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. A Rapid Method for Determining the Concentration of Recombinant Protein Secreted from Pichia pastoris

    Science.gov (United States)

    Sun, L. W.; Zhao, Y.; Niu, L. P.; Jiang, R.; Song, Y.; Feng, H.; feng, K.; Qi, C.

    2011-02-01

    Pichia secretive expression system is one of powerful eukaryotic expression systems in genetic engineering, which is especially suitable for industrial utilization. Because of the low concentration of the target protein in initial experiment, the methods and conditions for expression of the target protein should be optimized according to the protein yield repetitively. It is necessary to set up a rapid, simple and convenient analysis method for protein expression levels instead of the generally used method such as ultrafiltration, purification, dialysis, lyophilization and so on. In this paper, acetone precipitation method was chosen to concentrate the recombinant protein firstly after comparing with four different protein precipitation methods systematically, and then the protein was analyzed by SDS-Polyacrylamide Gel Electrophoresis. The recombinant protein was determined with the feature of protein band by the Automated Image Capture and 1-D Analysis Software directly. With this method, the optimized expression conditions of basic fibroblast growth factor secreted from pichia were obtained, which is as the same as using traditional methods. Hence, a convenient tool to determine the optimized conditions for the expression of recombinant proteins in Pichia was established.

  11. Aquifer configuration and geostructural links control the groundwater quality in thin-bedded carbonate–siliciclastic alternations of the Hainich CZE, central Germany

    Directory of Open Access Journals (Sweden)

    B. Kohlhepp

    2017-12-01

    Full Text Available The quality of near-surface groundwater reservoirs is controlled, but also threatened, by manifold surface–subsurface interactions. Vulnerability studies typically evaluate the variable interplay of surface factors (land management, infiltration patterns and subsurface factors (hydrostratigraphy, flow properties in a thorough way, but disregard the resulting groundwater quality. Conversely, hydrogeochemical case studies that address the chemical evolution of groundwater often lack a comprehensive analysis of the structural buildup. In this study, we aim to reconstruct the actual spatial groundwater quality pattern from a synoptic analysis of the hydrostratigraphy, lithostratigraphy, pedology and land use in the Hainich Critical Zone Exploratory (Hainich CZE. This CZE represents a widely distributed yet scarcely described setting of thin-bedded mixed carbonate–siliciclastic strata in hillslope terrains. At the eastern Hainich low-mountain hillslope, bedrock is mainly formed by alternated marine sedimentary rocks of the Upper Muschelkalk (Middle Triassic that partly host productive groundwater resources. Spatial patterns of the groundwater quality of a 5.4 km long well transect are derived by principal component analysis and hierarchical cluster analysis. Aquifer stratigraphy and geostructural links were deduced from lithological drill core analysis, mineralogical analysis, geophysical borehole logs and mapping data. Maps of preferential recharge zones and recharge potential were deduced from digital (soil mapping, soil survey data and field measurements of soil hydraulic conductivities (Ks. By attributing spatially variable surface and subsurface conditions, we were able to reconstruct groundwater quality clusters that reflect the type of land management in their preferential recharge areas, aquifer hydraulic conditions and cross-formational exchange via caprock sinkholes or ascending flow. Generally, the aquifer configuration (spatial

  12. Aquifer configuration and geostructural links control the groundwater quality in thin-bedded carbonate-siliciclastic alternations of the Hainich CZE, central Germany

    Science.gov (United States)

    Kohlhepp, Bernd; Lehmann, Robert; Seeber, Paul; Küsel, Kirsten; Trumbore, Susan E.; Totsche, Kai U.

    2017-12-01

    The quality of near-surface groundwater reservoirs is controlled, but also threatened, by manifold surface-subsurface interactions. Vulnerability studies typically evaluate the variable interplay of surface factors (land management, infiltration patterns) and subsurface factors (hydrostratigraphy, flow properties) in a thorough way, but disregard the resulting groundwater quality. Conversely, hydrogeochemical case studies that address the chemical evolution of groundwater often lack a comprehensive analysis of the structural buildup. In this study, we aim to reconstruct the actual spatial groundwater quality pattern from a synoptic analysis of the hydrostratigraphy, lithostratigraphy, pedology and land use in the Hainich Critical Zone Exploratory (Hainich CZE). This CZE represents a widely distributed yet scarcely described setting of thin-bedded mixed carbonate-siliciclastic strata in hillslope terrains. At the eastern Hainich low-mountain hillslope, bedrock is mainly formed by alternated marine sedimentary rocks of the Upper Muschelkalk (Middle Triassic) that partly host productive groundwater resources. Spatial patterns of the groundwater quality of a 5.4 km long well transect are derived by principal component analysis and hierarchical cluster analysis. Aquifer stratigraphy and geostructural links were deduced from lithological drill core analysis, mineralogical analysis, geophysical borehole logs and mapping data. Maps of preferential recharge zones and recharge potential were deduced from digital (soil) mapping, soil survey data and field measurements of soil hydraulic conductivities (Ks). By attributing spatially variable surface and subsurface conditions, we were able to reconstruct groundwater quality clusters that reflect the type of land management in their preferential recharge areas, aquifer hydraulic conditions and cross-formational exchange via caprock sinkholes or ascending flow. Generally, the aquifer configuration (spatial arrangement of strata

  13. Rapid Reticulin Fiber Staining Method is Helpful for the Diagnosis of Pituitary Adenoma in Frozen Section.

    Science.gov (United States)

    Noh, Songmi; Kim, Sun Ho; Cho, Nam Hoon; Kim, Se Hoon

    2015-05-01

    Approximately 90% of neoplasms found in the sellar region are adenoma of the pituitary gland. The use of frozen sections for the diagnosis of pituitary adenomas has an accuracy of 90% and is useful in evaluating complete tumor removal. However, it is sometimes difficult to diagnose pituitary adenomas using frozen sections because of the small sample size and marked artifact, and the contiguity of the pituitary adenoma with normal pituitary gland tissue. In this study, we evaluated the use of our modified reticulin stain to make correct decision in frozen section with reduced stain time and investigated the objective diagnostic criteria of pituitary adenoma with reticulin stain. We used Gomori's silver impregnation methods to stain reticulin fibers in frozen pituitary gland sections of 36 samples from 24 patients. We modified the conventional staining method by reducing the overall staining time. We diagnosed pituitary lesion according to our interpretation criteria and compared the results to those of the conventional method and findings of hematoxylin and eosin-stained slides. Reticulin fiber staining of normal adenohypophysis outlines the supporting stroma around the blood vessels and shows regular of the gland meshwork interconnecting the capillaries. In contrast, reticulin fiber staining of the adenomatous tissue shows loss of meshwork or frequent fragmentation. Our modified reticulin stain is more rapid than the established method and shows similar levels of accuracy. Independent evaluation by two pathologists showed discrepancies in diagnosis in four out of 36 cases with modified reticulin stain. Our rapid modified reticulin staining method for frozen sections may be useful as a diagnostic tool for pituitary adenomas and can complement routine hematoxylin and eosin staining.

  14. Clinical usefulness of multiplex PCR lateral flow in MRSA detection: a novel, rapid genetic testing method.

    Science.gov (United States)

    Nihonyanagi, Shin; Kanoh, Yuhsaku; Okada, Kiyomi; Uozumi, Toshiki; Kazuyama, Yukumasa; Yamaguchi, Tokiko; Nakazaki, Nobuhiko; Sakurai, Keizou; Hirata, Yasuyoshi; Munekata, Shinichi; Ohtani, Shinichi; Takemoto, Tsuyoshi; Bandoh, Yuki; Akahoshi, Tohru

    2012-06-01

    Methicillin-resistant Staphylococcus aureus (MRSA) with exogenous cassette DNA containing the methicillin-resistant gene mecA (SCCmec) poses a problem as a drug-resistant bacterium responsible for hospital- and community-acquired infections. The frequency of MRSA detection has recently been increasing rapidly in Japan, and SCCmec has also been classified more diversely into types I-V. A rapid test is essential for early diagnosis and treatment of MRSA infections, but detection by conventional methods requires at least two days. The newly developed multiplex PCR lateral flow method allows specific amplification of femA to detect S. aureus, mecA to detect SCCmec, and kdpC to detect SCCmec type II; moreover, PCR products can be evaluated visually in about 3 h. In the present study, we developed a PCR lateral flow method for MRSA using this method and investigated its clinical usefulness in the detection of MRSA. The results showed a diagnostic concordance rate of 91.7% for MRSA and methicillin-susceptible S. aureus between bacteriological examination and PCR lateral flow, and a high level of specificity in PCR lateral flow. In addition, a higher detection rate for S. aureus using the same sample was observed for PCR lateral flow (70.2%) than for bacteriological tests (48.6%). The above results show that PCR lateral flow for MRSA detection has high sensitivity, specificity, and speed, and its clinical application as a method for early diagnosis of MRSA infections appears to be feasible.

  15. Optimal estimation and scheduling in aquifer management using the rapid feedback control method

    Science.gov (United States)

    Ghorbanidehno, Hojat; Kokkinaki, Amalia; Kitanidis, Peter K.; Darve, Eric

    2017-12-01

    Management of water resources systems often involves a large number of parameters, as in the case of large, spatially heterogeneous aquifers, and a large number of "noisy" observations, as in the case of pressure observation in wells. Optimizing the operation of such systems requires both searching among many possible solutions and utilizing new information as it becomes available. However, the computational cost of this task increases rapidly with the size of the problem to the extent that textbook optimization methods are practically impossible to apply. In this paper, we present a new computationally efficient technique as a practical alternative for optimally operating large-scale dynamical systems. The proposed method, which we term Rapid Feedback Controller (RFC), provides a practical approach for combined monitoring, parameter estimation, uncertainty quantification, and optimal control for linear and nonlinear systems with a quadratic cost function. For illustration, we consider the case of a weakly nonlinear uncertain dynamical system with a quadratic objective function, specifically a two-dimensional heterogeneous aquifer management problem. To validate our method, we compare our results with the linear quadratic Gaussian (LQG) method, which is the basic approach for feedback control. We show that the computational cost of the RFC scales only linearly with the number of unknowns, a great improvement compared to the basic LQG control with a computational cost that scales quadratically. We demonstrate that the RFC method can obtain the optimal control values at a greatly reduced computational cost compared to the conventional LQG algorithm with small and controllable losses in the accuracy of the state and parameter estimation.

  16. Efficient 3D frequency response modeling with spectral accuracy by the rapid expansion method

    KAUST Repository

    Chu, Chunlei

    2012-07-01

    Frequency responses of seismic wave propagation can be obtained either by directly solving the frequency domain wave equations or by transforming the time domain wavefields using the Fourier transform. The former approach requires solving systems of linear equations, which becomes progressively difficult to tackle for larger scale models and for higher frequency components. On the contrary, the latter approach can be efficiently implemented using explicit time integration methods in conjunction with running summations as the computation progresses. Commonly used explicit time integration methods correspond to the truncated Taylor series approximations that can cause significant errors for large time steps. The rapid expansion method (REM) uses the Chebyshev expansion and offers an optimal solution to the second-order-in-time wave equations. When applying the Fourier transform to the time domain wavefield solution computed by the REM, we can derive a frequency response modeling formula that has the same form as the original time domain REM equation but with different summation coefficients. In particular, the summation coefficients for the frequency response modeling formula corresponds to the Fourier transform of those for the time domain modeling equation. As a result, we can directly compute frequency responses from the Chebyshev expansion polynomials rather than the time domain wavefield snapshots as do other time domain frequency response modeling methods. When combined with the pseudospectral method in space, this new frequency response modeling method can produce spectrally accurate results with high efficiency. © 2012 Society of Exploration Geophysicists.

  17. A new method for rapid determination of carbohydrate and total carbon concentrations using UV spectrophotometry.

    Science.gov (United States)

    Albalasmeh, Ammar A; Berhe, Asmeret Asefaw; Ghezzehei, Teamrat A

    2013-09-12

    A new UV spectrophotometry based method for determining the concentration and carbon content of carbohydrate solution was developed. This method depends on the inherent UV absorption potential of hydrolysis byproducts of carbohydrates formed by reaction with concentrated sulfuric acid (furfural derivatives). The proposed method is a major improvement over the widely used Phenol-Sulfuric Acid method developed by DuBois, Gilles, Hamilton, Rebers, and Smith (1956). In the old method, furfural is allowed to develop color by reaction with phenol and its concentration is detected by visible light absorption. Here we present a method that eliminates the coloration step and avoids the health and environmental hazards associated with phenol use. In addition, avoidance of this step was shown to improve measurement accuracy while significantly reducing waiting time prior to light absorption reading. The carbohydrates for which concentrations and carbon content can be reliably estimated with this new rapid Sulfuric Acid-UV technique include: monosaccharides, disaccharides and polysaccharides with very high molecular weight. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. A Simple and Rapid Data Extraction Method for the Precision Aspheric Optical Surface Height

    Science.gov (United States)

    Xing, Guohua; Peng, Yunfeng; Su, Xing

    2017-10-01

    Nowadays, the application of aspheric optics is becoming more and more popular in the precision optical engineering field. Therefore, it urges the rapid development of the precision machining and measuring technology. Generally, the aspheric optical component is measured by the interferometer. The underlying question is that the figure output by interferometer can’t be always recognized by other analysis software or program though the interferometer has its own unique data processing system. In this paper, a robust, rapid and simple method is presented to interpret the surface height data of the precision machined aspheric optical surface. The optical surface is measured by interferometer. The result figure is split into two parts, one of which is the interferogram picture of the whole aspheric optical surface and the other is the colour reference column indicating the height value. The ratios of the red (R), green (G) and blue (B) are analysed based on the middle of the colour reference column, and the corresponding relationship between the colours and surface height is established and looked as a reference data base. Then the interferogram picture of the whole aspheric optical surface is also analysed and divided according to the red (R), green (G) and blue (B) colours. By comparing the ratios and values of RGB colour, the aspheric optical surface height can be extracted approximately. The feasibility of this method was approved by the extraction processing experiment of a polished aspheric optical surface.

  19. A rapid, highly accurate method for quantifying CALR mutant allele burden in persons with myeloproliferative neoplasms.

    Science.gov (United States)

    Yao, Qiu-Mei; Zhou, Jiao; Gale, Robert Peter; Li, Jin-Lan; Li, Ling-Di; Li, Ning; Chen, Shan-Shan; Ruan, Guo-Rui

    2015-10-01

    Calreticulin (CALR) mutations were recently identified in a substantial proportion of persons with essential thrombocythemia (ET) and with primary myelofibrosis (PMF) without JAK2(V617F). Consequently rapid, sensitive, and specific methods to detect and quantify these mutations are needed. We studied samples from 1088 persons with myeloproliferative neoplasms (MPNs) including 421 JAK2(V617F) negative subjects with ET, PMF, polycythemia vera (PV), chronic myeloid leukemia (CML) and hyper-eosinophilic syndrome (HES). Detection of CALR exon 9 mutations was done by PCR amplification followed by fragment length analysis and direct sequencing. Dilution assays were used to determine CALR mutant allele burden. We detected CALR mutations in blood and bone marrow samples from 152 subjects with ET and with PMF but not in samples from normal or persons with PV, CML, or HES. CALR mutant peaks were distinct from wild-type peaks and dilution experiments indicated a sensitivity level of 0.5-5% for a CALR mutant allele in a wild-type background. Diverse types of mutations were detected including deletions, insertions, and complex indels. All mutations were confirmed by direct sequencing. We also used dilution experiments to quantify mutant allele burden. We were able to reproducibly detect mutant allele levels as low 5% (0.5-5%) in a wild-type background. PCR amplification followed by fragment length analysis is a rapid, sensitive, and specific method for screening persons with MPNs for CALR mutations, especially those with ET and PMF and for estimating mutant allele burden.

  20. Rapid fabrication method of a microneedle mold with controllable needle height and width.

    Science.gov (United States)

    Lin, Yen-Heng; Lee, I-Chi; Hsu, Wei-Chieh; Hsu, Ching-Hong; Chang, Kai-Ping; Gao, Shao-Syuan

    2016-10-01

    The main issue of transdermal drug delivery is that macromolecular drugs cannot diffuse through the stratum corneum of skin. Many studies have pursued micro-sized needles encapsulated with drugs to overcome this problem, as these needles can pierce the stratum corneum and allow drugs to enter the circulatory system of the human body. However, most microneedle fabrication processes are time-consuming and require expensive equipment. In this study, we demonstrate a rapid method for fabricating a microneedle mold using drawing lithography and a UV-cured resin. The mold was filled with a water-soluble material, polyvinylpyrrolidone (PVP), which was then demolded to produce a water-soluble microneedle array. The results of an in vitro skin insertion test using PVP microneedles and pig ear skin demonstrated the feasibility of the microneedle mold. In addition, by controlling the viscosity of the UV-cured resin through various heat treatments, microneedles with different heights and aspect ratios were produced. Compared with other methods, this technology significantly simplifies and accelerates the mold fabrication process. In addition, the required equipment is relatively simple and inexpensive. Through this technology, we can rapidly fabricate microneedle molds with controllable dimensions for various applications.

  1. Passive acoustic methods for fine-scale tracking of harbour porpoises in tidal rapids.

    Science.gov (United States)

    Macaulay, Jamie; Gordon, Jonathan; Gillespie, Douglas; Malinka, Chloë; Northridge, Simon

    2017-02-01

    The growing interest in generating electrical power from tidal currents using tidal turbine generators raises a number of environmental concerns, including the risk that marine mammals might be injured or killed through collision with rotating turbine blades. To understand this risk, information on how marine mammals use tidal rapid habitats and in particular, their underwater movements and dive behaviour is required. Porpoises, which are the most abundant small cetacean at most European tidal sites, are difficult animals to tag, and the limited size of tidal habitats means that any telemetered animal would be likely to spend only a small proportion of time within them. Here, an alternative approach is explored, whereby passive acoustic monitoring (PAM) is used to obtain fine scale geo-referenced tracks of harbour porpoises in tidal rapid areas. Large aperture hydrophone arrays are required to obtain accurate locations of animals from PAM data and automated algorithms are necessary to process the large quantities of acoustic data collected on such systems during a typical survey. Methods to automate localisation, including a method to match porpoise detections on different hydrophones and separate different vocalising animals, and an assessment of the localisation accuracy of the large aperture hydrophone array are presented.

  2. Miniaturized fluorescent RNA dot blot method for rapid quantitation of gene expression

    Directory of Open Access Journals (Sweden)

    Yadetie Fekadu

    2004-06-01

    Full Text Available Abstract Background RNA dot blot hybridization is a commonly used technique for gene expression assays. However, membrane based RNA dot/slot blot hybridization is time consuming, requires large amounts of RNA, and is less suited for parallel assays of more than one gene at a time. Here, we describe a glass-slide based miniaturized RNA dot blot (RNA array procedure for rapid and parallel gene expression analysis using fluorescently labeled probes. Results RNA arrays were prepared by simple manual spotting of RNA onto amino-silane coated microarray glass slides, and used for two-color fluorescent hybridization with specific probes labeled with Cy3 and 18S ribosomal RNA house-keeping gene probe labeled with Cy5 fluorescent dyes. After hybridization, arrays were scanned on a fluorescent microarray scanner and images analyzed using microarray image analysis software. We demonstrate that this method gives comparable results to Northern blot analysis, and enables high throughput quantification of transcripts from nanogram quantities of total RNA in hundreds of samples. Conclusion RNA array on glass slide and detection by fluorescently labeled probes can be used for rapid and parallel gene expression analysis. The method is particularly well suited for gene expression assays that involve quantitation of many transcripts in large numbers of samples.

  3. Hyperspectral Imaging as a Rapid Quality Control Method for Herbal Tea Blends

    Directory of Open Access Journals (Sweden)

    Majolie Djokam

    2017-03-01

    Full Text Available In South Africa, indigenous herbal teas are enjoyed due to their distinct taste and aroma. The acclaimed health benefits of herbal teas include the management of chronic diseases such as hypertension and diabetes. Quality control of herbal teas has become important due to the availability of different brands of varying quality and the production of tea blends. The potential of hyperspectral imaging as a rapid quality control method for herbal tea blends from rooibos (Aspalathus linearis, honeybush (Cyclopia intermedia, buchu (Agathosma Betulina and cancerbush (Sutherlandia frutescens was investigated. Hyperspectral images of raw materials and intact tea bags were acquired using a sisuChema shortwave infrared (SWIR hyperspectral pushbroom imaging system (920–2514 nm. Principal component analysis (PCA plots showed clear discrimination between raw materials. Partial least squares discriminant analysis (PLS-DA models correctly predicted the raw material constituents of each blend and accurately determined the relative proportions. The results were corroborated independently using ultra-high performance liquid chromatography coupled to mass spectrometry (UHPLC-MS. This study demonstrated the application of hyperspectral imaging coupled with chemometric modelling as a reliable, rapid and non-destructive quality control method for authenticating herbal tea blends and to determine relative proportions in a tea bag.

  4. Rapid and label-free bioanalytical method of alpha fetoprotein detection using LSPR chip

    Science.gov (United States)

    Kim, Dongjoo; Kim, Jinwoon; Kwak, Cheol Hwan; Heo, Nam Su; Oh, Seo Yeong; Lee, Hoomin; Lee, Go-Woon; Vilian, A. T. Ezhil; Han, Young-Kyu; Kim, Woo-Sik; Kim, Gi-bum; Kwon, Soonjo; Huh, Yun Suk

    2017-07-01

    Alpha fetoprotein (AFP) is a cancer marker, particularly for hepatocellular carcinoma. Normal levels of AFP are less than 20 ng/mL; however, its levels can reach more than 400 ng/mL in patients with HCC. Enzyme linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) have been employed for clinical diagnosis of AFP; however, these methods are time consuming and labor intensive. In this study, we developed a localized surface plasmon resonance (LSPR) based biosensor for simple and rapid detection of AFP. This biosensor consists of a UV-Vis spectrometer, a cuvette cell, and a biosensor chip nanopatterned with gold nanoparticles (AuNPs). In our LSPR biosensor, binding of AFP to the surface of the sensor chip led to an increasing magnitude of the LSPR signals, which was measured by an ultraviolet-visible (UV-Vis) spectrometer. Our LSPR biosensor showed sufficient detectability of AFP at concentrations of 1 ng/mL to 1 μg/mL. Moreover, the overall procedure for detection of AFP was completed within 20 min. This biosensor could also be utilized for a point of care test (POCT) by employing a portable UV-Vis spectrometer. Owing to the simplicity and rapidity of the detection process, our LSPR biosensor is expected to replace traditional diagnostic methods for the early detection of diseases.

  5. Rapid HPLC method for the simultaneous monitoring of duloxetine, venlaflaxine, fluoxetine and paroxetine in biofluids.

    Science.gov (United States)

    Samanidou, Victoria F; Kourti, Paraskevi V

    2009-08-01

    A simple and rapid HPLC method is developed for the determination of two serotonin-norepinephrine-reuptake inhibitors (duloxetine and venlaflaxine) and two selective serotonin-reuptake inhibitors (fluoxetine and paroxetine) in human biofluids. Separation was performed on an Inertsil ODS-3 column (250 x 4.0 mm, 5 µm) with acetonitrile-ammonium acetate (0.05 M, 41:59 v/v) at 235 nm, within 7 min. SPE on Oasis(®) HLB cartridges was applied for the isolation of analytes from biofluids. The developed methodology was validated in terms of sensitivity, linearity, accuracy, precision, stability and selectivity. Relative standard deviation was less than 10.4%. Limit of detection was 0.2-0.6 ng/µl in blood plasma and 0.1-0.8 ng/µl in urine. The method was successfully applied to biofluids from a patient under duloxetine treatment.

  6. A rapid and scalable method for selecting recombinant mouse monoclonal antibodies

    Directory of Open Access Journals (Sweden)

    Wright Gavin J

    2010-06-01

    Full Text Available Abstract Background Monoclonal antibodies with high affinity and selectivity that work on wholemount fixed tissues are valuable reagents to the cell and developmental biologist, and yet isolating them remains a long and unpredictable process. Here we report a rapid and scalable method to select and express recombinant mouse monoclonal antibodies that are essentially equivalent to those secreted by parental IgG-isotype hybridomas. Results Increased throughput was achieved by immunizing mice with pools of antigens and cloning - from small numbers of hybridoma cells - the functionally rearranged light and heavy chains into a single expression plasmid. By immunizing with the ectodomains of zebrafish cell surface receptor proteins expressed in mammalian cells and screening for formalin-resistant epitopes, we selected antibodies that gave expected staining patterns on wholemount fixed zebrafish embryos. Conclusions This method can be used to quickly select several high quality monoclonal antibodies from a single immunized mouse and facilitates their distribution using plasmids.

  7. A method for rapidly screening functionality of actin mutants and tagged actins

    Directory of Open Access Journals (Sweden)

    Rommelaere Heidi

    2004-01-01

    Full Text Available Recombinant production and biochemical analysis of actin mutants has been hampered by the fact that actin has an absolute requirement for the eukaryotic chaperone CCT to reach its native state. We therefore have developed a method to rapidly screen the folding capacity and functionality of actin variants, by combining in vitro expression of labelled actin with analysis on native gels, band shift assays or copolymerization tests. Additionally, we monitor, using immuno-fluorescence, incorporation of actin variants in cytoskeletal structures in transfected cells. We illustrate the method by two examples. In one we show that tagged versions of actin do not always behave native-like and in the other we study some of the molecular defects of three &bgr;-actin mutants that have been associated with diseases.

  8. A rapid and efficient DNA extraction method suitable for marine macroalgae.

    Science.gov (United States)

    Ramakrishnan, Gautham Subramaniam; Fathima, Anwar Aliya; Ramya, Mohandass

    2017-12-01

    Macroalgae are a diverse group of organisms. Marine macroalgae, in particular, have numerous medicinal and industrial applications. Molecular studies of macroalgae require suitable concentrations of DNA free of contaminants. At present, numerous protocols exist for DNA extraction from macroalgae. However, they are either time consuming, expensive or work only with few species. The method described in this study is rapid and efficient and applicable to different types of marine macroalgae. This method yields an average of 3.85 µg of DNA per 50 mg of algal tissue, with an average purity of 1.88. The isolated DNA was suitable for PCR amplification of universal plastid region of macroalgae.

  9. A Rapid Colorimetric Method Reveals Fraudulent Substitutions in Sea Urchin Roe Marketed in Sardinia (Italy).

    Science.gov (United States)

    Meloni, Domenico; Spina, Antonio; Satta, Gianluca; Chessa, Vittorio

    2016-06-25

    In recent years, besides the consumption of fresh sea urchin specimens, the demand of minimally-processed roe has grown considerably. This product has made frequent consumption in restaurants possible and frauds are becoming widespread with the partial replacement of sea urchin roe with surrogates that are similar in colour. One of the main factors that determines the quality of the roe is its colour and small differences in colour scale cannot be easily discerned by the consumers. In this study we have applied a rapid colorimetric method for reveal the fraudulent partial substitution of semi-solid sea urchin roe with liquid egg yolk. Objective assessment of whiteness (L*), redness (a*), yellowness (b*), hue (h*), and chroma (C*) was carried out with a digital spectrophotometer using the CIE L*a*b* colour measurement system. The colorimetric method highlighted statistically significant differences among sea urchin roe and liquid egg yolk that could be easily discerned quantitatively.

  10. Meselect – A rapid and effective method for the separation of the main leaf tissue types

    Directory of Open Access Journals (Sweden)

    Julia Svozil

    2016-11-01

    Full Text Available Individual tissues of complex eukaryotic organisms have specific gene expression programs that control their functions. Therefore, tissue-specific molecular information is required to increase our understanding of tissue-specific processes. Established methods in plants to obtain specific tissues or cell types from their organ or tissue context typically require the enzymatic degradation of cell walls followed by fluorescence-activated cell sorting (FACS using plants engineered for localized expression of green fluorescent protein (GFP. This has facilitated the acquisition of valuable data, mainly on root cell type-specific transcript and protein expression. However, FACS of different leaf cell types is difficult because of chlorophyll autofluorescence that interferes with the sorting process. Furthermore, the cell wall composition is different in each cell type. This results in long incubation times for refractory cell types, and cell sorting itself can take several hours. To overcome these limitations, we developed Meselect (mechanical separation of leaf compound tissues, a rapid and effective method for the separation of leaf epidermal, vascular and mesophyll tissues. Meselect is a novel combination of mechanical separation and rapid protoplasting, which benefits from the unique cell wall composition of the different tissue types. Meselect has several advantages over cell sorting: it does not require expensive equipment such as a cell sorter and does not depend on specific fluorescent reporter lines, the use of blenders as well as the inherent mixing of different cell types and of intact and damaged cells can be avoided, and the time between wounding of the leaf and freezing of the sample is short. The efficacy and specificity of the method to enrich the different leaf tissue types has been confirmed using Arabidopsis leaves, but it has also been successfully used for leaves of other plants such as tomato or cassava. The method is therefore

  11. Development of rapid methods for relaxation time mapping and motion estimation using magnetic resonance imaging

    Energy Technology Data Exchange (ETDEWEB)

    Gilani, Syed Irtiza Ali

    2008-09-15

    Recent technological developments in the field of magnetic resonance imaging have resulted in advanced techniques that can reduce the total time to acquire images. For applications such as relaxation time mapping, which enables improved visualisation of in vivo structures, rapid imaging techniques are highly desirable. TAPIR is a Look- Locker-based sequence for high-resolution, multislice T{sub 1} relaxation time mapping. Despite the high accuracy and precision of TAPIR, an improvement in the k-space sampling trajectory is desired to acquire data in clinically acceptable times. In this thesis, a new trajectory, termed line-sharing, is introduced for TAPIR that can potentially reduce the acquisition time by 40 %. Additionally, the line-sharing method was compared with the GRAPPA parallel imaging method. These methods were employed to reconstruct time-point images from the data acquired on a 4T high-field MR research scanner. Multislice, multipoint in vivo results obtained using these methods are presented. Despite improvement in acquisition speed, through line-sharing, for example, motion remains a problem and artefact-free data cannot always be obtained. Therefore, in this thesis, a rapid technique is introduced to estimate in-plane motion. The presented technique is based on calculating the in-plane motion parameters, i.e., translation and rotation, by registering the low-resolution MR images. The rotation estimation method is based on the pseudo-polar FFT, where the Fourier domain is composed of frequencies that reside in an oversampled set of non-angularly, equispaced points. The essence of the method is that unlike other Fourier-based registration schemes, the employed approach does not require any interpolation to calculate the pseudo-polar FFT grid coordinates. Translation parameters are estimated by the phase correlation method. However, instead of two-dimensional analysis of the phase correlation matrix, a low complexity subspace identification of the phase

  12. Autoclave method for rapid preparation of bacterial PCR-template DNA.

    Science.gov (United States)

    Simmon, Keith E; Steadman, Dewey D; Durkin, Sarah; Baldwin, Amy; Jeffrey, Wade H; Sheridan, Peter; Horton, Rene; Shields, Malcolm S

    2004-02-01

    An autoclave method for preparing bacterial DNA for PCR template is presented, it eliminates the use of detergents, organic solvents, and mechanical cellular disruption approaches, thereby significantly reducing processing time and costs while increasing reproducibility. Bacteria are lysed by rapid heating and depressurization in an autoclave. The lysate, cleared by microcentrifugation, was either used directly in the PCR reaction, or concentrated by ultrafiltration. This approach was compared with seven established methods of DNA template preparation from four bacterial sources which included boiling Triton X-100 and SDS, bead beating, lysozyme/proteinase K, and CTAB lysis method components. Bacteria examined were Enterococcus and Escherichia coli, a natural marine bacterial community and an Antarctic cyanobacterial-mat. DNAs were tested for their suitability as PCR templates by repetitive element random amplified polymorphic DNA (RAPD) and denaturing gradient gel electrophoresis (DGGE) analysis. The autoclave method produced PCR amplifiable template comparable or superior to the other methods, with greater reproducibility, much shorter processing time, and at a significantly lower cost.

  13. A tree-based method for the rapid screening of chemical fingerprints

    Directory of Open Access Journals (Sweden)

    Pedersen Christian NS

    2010-01-01

    Full Text Available Abstract Background The fingerprint of a molecule is a bitstring based on its structure, constructed such that structurally similar molecules will have similar fingerprints. Molecular fingerprints can be used in an initial phase of drug development for identifying novel drug candidates by screening large databases for molecules with fingerprints similar to a query fingerprint. Results In this paper, we present a method which efficiently finds all fingerprints in a database with Tanimoto coefficient to the query fingerprint above a user defined threshold. The method is based on two novel data structures for rapid screening of large databases: the kD grid and the Multibit tree. The kD grid is based on splitting the fingerprints into k shorter bitstrings and utilising these to compute bounds on the similarity of the complete bitstrings. The Multibit tree uses hierarchical clustering and similarity within each cluster to compute similar bounds. We have implemented our method and tested it on a large real-world data set. Our experiments show that our method yields approximately a three-fold speed-up over previous methods. Conclusions Using the novel kD grid and Multibit tree significantly reduce the time needed for searching databases of fingerprints. This will allow researchers to (1 perform more searches than previously possible and (2 to easily search large databases.

  14. Development of a micropulverized extraction method for rapid toxicological analysis of methamphetamine in hair.

    Science.gov (United States)

    Miyaguchi, Hajime; Kakuta, Masaya; Iwata, Yuko T; Matsuda, Hideaki; Tazawa, Hidekatsu; Kimura, Hiroko; Inoue, Hiroyuki

    2007-09-07

    We developed a rapid sample preparation method for the toxicological analysis of methamphetamine and amphetamine (the major metabolite of methamphetamine) in human hair by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), to facilitate fast screening and quantitation. Two milligrams of hair were mechanically micropulverized for 5 min in a 2-ml plastic tube together with 100 microl of an aqueous solvent containing 10% acetonitrile, 100 mM trifluoroacetic acid and the corresponding deuterium analogues as internal standards. The pulverizing highly disintegrated the hair components, simultaneously allowing the extraction of any drugs present in the hair. After filtering the suspension with a membrane-filter unit, the clear filtrate was directly analyzed by HPLC-MS/MS. No evaporation processes were required for sample preparation. Method optimization and validation study were carried out using real-case specimens and fortified samples in which the drugs had been artificially absorbed, respectively. Concentration ranges for quantitation were 0.040-125 and 0.040-25 ng/mg for methamphetamine and amphetamine, respectively. Real-case specimens were analyzed by the method presented here and by conventional ones to verify the applicability of our method to real-world analysis. Our method took less than 30 min for a set of chromatograms to be obtained from a washed hair sample.

  15. Zastosowanie hormonu wzrostu w leczeniu zaburzeń wzrastania u dzieci chorych na młodzieńcze idiopatyczne zapalenie stawów

    Directory of Open Access Journals (Sweden)

    Jacek Postępski

    2010-04-01

    Full Text Available Zaburzenia wzrastania stanowią bardzo istotny problem u dzieciz młodzieńczym idiopatycznym zapaleniem stawów (MIZS, dotycząone zwłaszcza pacjentów z postacią o początku uogólnionymi wielostawowym.W pracy na podstawie piśmiennictwa przedstawiono przeglądwyników badań klinicznych i obserwacyjnych dotyczących ocenyzaburzeń wzrastania i leczenia hormonem wzrostu dzieci chorychna młodzieńcze idiopatyczne zapalenie stawów.Zahamowanie wzrostu w przebiegu MIZS jest związane główniez istniejącym procesem zapalnym i nasilone przez niepożądanedziałanie glikokortykosteroidów. Nowoczesne metody leczenia niezawsze zapobiegają niedoborowi wzrostu, wskazane jest wówczaszastosowanie hormonu wzrostu. Najlepsze rezultaty przynosizastosowanie hormonu wzrostu we wczesnej fazie choroby,zanim dojdzie do ciężkich niedoborów wzrostu.

  16. NATO Advanced Research Workshop, 19-22 May 1997: Rapid Method for Monitoring the Environment for Biological Hazards.

    Science.gov (United States)

    1997-05-22

    The NATO Advanced Research Workshop met for the purpose of bringing to light rapid methods for monitoring the environment for biological hazards such as biological warfare agents, naturally occurring diseases, detection and identification of different biological threats in the environment , dormancy in non-sporulating bacteria and bioluminescence techniques with respect to the rapid detection of microbes in air, water and food.

  17. Rapid Assessment of Health Services in Punjab using a Mixed Method Approach

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    Rajesh Kumar

    2015-06-01

    Full Text Available Introduction: The out-of-pocket expenditure is quite high in Punjab. Hence, a rapid review of health facilities was undertaken to suggest remedial measures. Methods: Mixed method research approach was used to identify strengths and weaknesses of the health services in Punjab. All health institutions were included in the assessment from the three purposively sampled districts – one from each of the three regions of Punjab. Tools were developed to collect data from record review, observations, and in-depth interviews. Six building blocks framework proposed by the World Health Organization was used for data collection and analyses. Results: In general physical infrastructure, especially the buildings were found to be reasonably constructed at most of the healthcare facilities. However, the maintenance was not regular. The vacancies for general doctors, specialist doctors, nurses, and paramedics were 26%, 38%, 31% and 12% respectively. Supply of drugs was irregular and inadequate. A large proportion (45% of ‘user charges’ were spent on purchase of drugs and other consumables. Most registers were found to be updated, and reports were transmitted to higher levels usually on time. However, institutionalized system of monitoring and supervision was lacking. Govt. hospitals were providing in-patient care to about 35.5% of those who were estimated to need hospitalization. State had allocated about Rs. 1200 crores to health (0.46% of GDP, thus, spending only Rs. 433 per capita per year. Conclusions: Despite constraints, the government health service is catering to the needs of a large section of the population. Rapid health system assessment at periodic intervals using a mixed method approach can supplement routine monitoring of the health system.

  18. A rapid method of detecting autoantibody against FcεRIα for chronic spontaneous urticaria.

    Directory of Open Access Journals (Sweden)

    Mey-Fann Lee

    Full Text Available BACKGROUND: Chronic spontaneous urticaria (CU is a common skin disorder, with an estimated prevalence of 0.5-1.8% in most populations. Around 30-50% of CU patients have an autoimmune etiology, with autoantibodies (autoAbs against IgE, FcεRIα, and FcεRII/CD23. Although the in vivo autologous serum skin test (ASST and in vitro histamine release/activation assay are the most frequently used screening methods, these two have many limitations and do not directly measure susceptible autoAbs. This study aimed to establish an in vitro rapid screening test using recombinant autoantigen FcεRIα(rFcεRIα to improve the diagnosis of autoimmune urticaria. METHODS: Forty patients with CU and 20 healthy individuals were enrolled. After PCR-based cloning and the production of extracellular fragments of the FcεRIα protein using the E. coli expression system, serum autoAb to rFcεRIα was evaluated using in-house ELISA and rapid immunodot test. RESULTS: In ELISA-based detection, 14 out of 20 CU-ASST(+ patients exhibited anti- FcεRIα responses, whereas five of the 20 CU-ASST(- and two of the 20 non-CU patients showed autoantibody background in the assay. For the immunodot test, 55% (11/20 of the CU-ASST(+ sera exhibited anti-FcεRIα reactivity. There was no false positive among the CU-ASST(- and non-CU groups. Using clinical urticaria plus ASST(+ as the gold standard, in-house ELISA had 70% sensitivity, 82.5% specificity, and positive likelihood ratio of 4, while immunodot had 55% sensitivity, 100% specificity, and positive likelihood ratio >55. CONCLUSIONS: This study has developed a rapid immunodot method with high specificity for detecting autoAb to FcεRIαin patients with CU. Preliminary data indicates that this immunodot technique has the potential to be a routine diagnostic assay for autoimmune CU.

  19. Rapid Hydrothermal Synthesis of Zinc Oxide Nanowires by Annealing Methods on Seed Layers

    Directory of Open Access Journals (Sweden)

    Jang Bo Shim

    2011-01-01

    Full Text Available Well-aligned zinc oxide (ZnO nanowire arrays were successfully synthesized on a glass substrate using the rapid microwave heating process. The ZnO seed layers were produced by spinning the precursor solutions onto the substrate. Among coatings, the ZnO seed layers were annealed at 100°C for 5 minutes to ensure particle adhesion to the glass surface in air, nitrogen, and vacuum atmospheres. The annealing treatment of the ZnO seed layer was most important for achieving the high quality of ZnO nanowire arrays as ZnO seed nanoparticles of larger than 30 nm in diameter evolve into ZnO nanowire arrays. Transmission electron microscopy analysis revealed a single-crystalline lattice of the ZnO nanowires. Because of their low power (140 W, low operating temperatures (90°C, easy fabrication (variable microwave sintering system, and low cost (90% cost reduction compared with gas condensation methods, high quality ZnO nanowires created with the rapid microwave heating process show great promise for use in flexible solar cells and flexible display devices.

  20. A Rapid and Efficient Method for Evaluation of Suspect Testimony: Palynological Scanning.

    Science.gov (United States)

    Wiltshire, Patricia E J; Hawksworth, David L; Edwards, Kevin J

    2015-11-01

    A rapid method for evaluating suspect testimony is valuable at any stage in an inquiry and can result in a change of direction in an investigation. Rape cases, in particular, can present problems where a defendant renders DNA analysis redundant by claiming that the claimant consented to have sexual relations. Forensic palynology is valuable in confirming or eliminating locations as being crime scenes, thus checking the testimony of both parties. In contrast to some forensic disciplines, forensic palynology can provide critical information without time-consuming full analysis. Two cases are described where the palynological assemblages from comparator samples of pertinent places were compared with those obtained from clothing of claimants and defendants. The results of rapid microscopical scanning of relevant preparations led to early confessions, thus obviating the need for costly analyses and protracted court proceedings. A third case demonstrates the unbiased nature of this technique where a man, although innocent of any offense, lied about having visited the crime scene for fear of prosecution. This highlights the need for sensitive policing in claims of rape. © 2015 American Academy of Forensic Sciences.

  1. Rapid and Sensitive Lateral Flow Immunoassay Method for Procalcitonin (PCT Based on Time-Resolved Immunochromatography

    Directory of Open Access Journals (Sweden)

    Xiang-Yang Shao

    2017-02-01

    Full Text Available Procalcitonin (PCT is a current, frequently-used marker for severe bacterial infection. The aim of this study was to develop a cost-effective detection kit for rapid quantitative and on-site detection of PCT. To develop the new PCT quantitative detecting kit, a double-antibody sandwich immunofluorescent assay was employed based on time-resolved immunofluorescent assay (TRFIA combined with lateral flow immunoassay (LFIA. The performance of the new developed kit was evaluated in the aspects of linearity, precision, accuracy, and specificity. Two-hundred thirty-four serum samples were enrolled to carry out the comparison test. The new PCT quantitative detecting kit exhibited a higher sensitivity (0.08 ng/mL. The inter-assay coefficient of variation (CV and the intra-assay CV were 5.4%–7.7% and 5.7%–13.4%, respectively. The recovery rates ranged from 93% to 105%. Furthermore, a high correlation (n = 234, r = 0.977, p < 0.0001 and consistency (Kappa = 0.875 were obtained when compared with the PCT kit from Roche Elecsys BRAHMS. Thus, the new quantitative method for detecting PCT has been successfully established. The results indicated that the newly-developed system based on TRFIA combined with LFIA was suitable for rapid and on-site detection for PCT, which might be a useful platform for other biomarkers in point-of-care tests.

  2. Bacterial Cytological Profiling (BCP as a Rapid and Accurate Antimicrobial Susceptibility Testing Method for Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    D.T. Quach

    2016-02-01

    Full Text Available Successful treatment of bacterial infections requires the timely administration of appropriate antimicrobial therapy. The failure to initiate the correct therapy in a timely fashion results in poor clinical outcomes, longer hospital stays, and higher medical costs. Current approaches to antibiotic susceptibility testing of cultured pathogens have key limitations ranging from long run times to dependence on prior knowledge of genetic mechanisms of resistance. We have developed a rapid antimicrobial susceptibility assay for Staphylococcus aureus based on bacterial cytological profiling (BCP, which uses quantitative fluorescence microscopy to measure antibiotic induced changes in cellular architecture. BCP discriminated between methicillin-susceptible (MSSA and -resistant (MRSA clinical isolates of S. aureus (n = 71 within 1–2 h with 100% accuracy. Similarly, BCP correctly distinguished daptomycin susceptible (DS from daptomycin non-susceptible (DNS S. aureus strains (n = 20 within 30 min. Among MRSA isolates, BCP further identified two classes of strains that differ in their susceptibility to specific combinations of beta-lactam antibiotics. BCP provides a rapid and flexible alternative to gene-based susceptibility testing methods for S. aureus, and should be readily adaptable to different antibiotics and bacterial species as new mechanisms of resistance or multidrug-resistant pathogens evolve and appear in mainstream clinical practice.

  3. OSO paradigm--A rapid behavioral screening method for acute psychosocial stress reactivity in mice.

    Science.gov (United States)

    Brzózka, M M; Unterbarnscheidt, T; Schwab, M H; Rossner, M J

    2016-02-09

    Chronic psychosocial stress is an important environmental risk factor for the development of psychiatric diseases. However, studying the impact of chronic psychosocial stress in mice is time consuming and thus not optimally suited to 'screen' increasing numbers of genetically manipulated mouse models for psychiatric endophenotypes. Moreover, many studies focus on restraint stress, a strong physical stressor with limited relevance for psychiatric disorders. Here, we describe a simple and a rapid method based on the resident-intruder paradigm to examine acute effects of mild psychosocial stress in mice. The OSO paradigm (open field--social defeat--open field) compares behavioral consequences on locomotor activity, anxiety and curiosity before and after exposure to acute social defeat stress. We first evaluated OSO in male C57Bl/6 wildtype mice where a single episode of social defeat reduced locomotor activity, increased anxiety and diminished exploratory behavior. Subsequently, we applied the OSO paradigm to mouse models of two schizophrenia (SZ) risk genes. Transgenic mice with neuronal overexpression of Neuregulin-1 (Nrg1) type III showed increased risk-taking behavior after acute stress exposure suggesting that NRG1 dysfunction is associated with altered affective behavior. In contrast, Tcf4 transgenic mice displayed a normal stress response which is in line with the postulated predominant contribution of TCF4 to cognitive deficits of SZ. In conclusion, the OSO paradigm allows for rapid screening of selected psychosocial stress-induced behavioral endophenotypes in mouse models of psychiatric diseases. Copyright © 2015 IBRO. Published by Elsevier Ltd. All rights reserved.

  4. Performance of a Micro-UAV lifting system built with the usage of rapid prototyping methods

    Science.gov (United States)

    Dalewski, R. T.; Gumowski, K.; Barczak, T.; Godek, J.

    2014-08-01

    This article presents results of the aerodynamic testing of a micro unmanned aerial vehicle rotor efficiency. The rotors were prepared as a set of two rotors in a counter-rotating ducted drive. Prototypes of the drives were made using two rapid prototyping techniques - FDM - fused deposition modelling method and SLS - selective laser sintering. Rotors were made then treated by introducing additional finishing cyanoacrylate coating and abrasive processing. Main differences between those models were observed in fan shape, porosity, surface roughness and mechanical properties - stiffness. An influence of these factors was observed on an aerodynamic efficiency. For the obtained prototypes both simulations and experimental testing were conducted with thrust, power, torque measurements, as well as the measurement of velocity and pressure distribution at the outlet of the duct. The results show the possibility of using rapid prototyping techniques to produce prototypes of drives operating in the low and medium Reynolds numbers (6000-60000), and the aerodynamic shape relevant factors affecting the preparation and performance of such drives. In addition, simulation studies were performed using the Fluent environment where experimental results were confronted with the results of simulation studies.

  5. Use of a modified cluster sampling method to perform rapid needs assessment after Hurricane Andrew.

    Science.gov (United States)

    Hlady, W G; Quenemoen, L E; Armenia-Cope, R R; Hurt, K J; Malilay, J; Noji, E K; Wurm, G

    1994-04-01

    To rapidly obtain population-based estimates of needs in the early aftermath of Hurricane Andrew in South Florida. We used a modified cluster-sampling method (the Expanded Programme on Immunization [EPI] method) for three surveys. We selected a systematic sample of 30 quarter-mile square clusters for each survey and, beginning from a random start, interviewed members of seven consecutive occupied households in each cluster. Two surveys were of the most affected area (1990 population, 32,672) at three and ten days after the hurricane struck; one survey was of a less affected area (1990 population, 15,576) seven days after the hurricane struck. Results were available within 24 hours of beginning each survey. Initial findings emphasized the need for restoring utilities and sanitation and helped to focus medical relief on primary care and preventive services. The second survey of the most affected area showed improvement in the availability of food, water, electricity, and sanitation (P < or = .05). There was no evidence of disease outbreaks. For the first time, the EPI method provided population-based information to guide and evaluate relief operations after a sudden-impact natural disaster. An improvement over previous approaches, the EPI method warrants further evaluation as a needs assessment tool in acute disasters.

  6. ATP bioluminescence method: tool for rapid screening of organic and microbial contaminants on deteriorated mural paintings.

    Science.gov (United States)

    Unković, Nikola; Ljaljević Grbić, Milica; Stupar, Miloš; Vukojević, Jelena; Subakov-Simić, Gordana; Jelikić, Aleksa; Stanojević, Dragan

    2015-11-24

    The extent of the microbial contamination of the seventeenth-century wall paintings in the nave of the old Church of the Holy Ascension (Veliki Krčimir, Serbia) was evaluated via newly implemented ATP bioluminescence method, and traditional cultivation-based method, utilising commercially available dip slides. To assess the validity of ATP, as a biomarker for rapid detection of mural surface contamination, obtained zones of cleanliness values, in range from 1.0 to 5.3, were compared to documented total microbial counts, ranging between seven and 247 CFU/cm 2 . Small coefficients of determination, 0.0106-0.0385, suggest poor correlation between microbial counts and surface ATP levels; however, zones of cleanliness values are of great help in determining the high points of contamination, aka 'hotspots', which should be given special attention during sampling and investigation using other methods. In addition, various aspects of the possible implementation of the ATP bioluminescence method in an integrated system of wall painting conservation are discussed.

  7. A Rapid Dialysis Method for Analysis of Artificial Sweeteners in Foods (2nd Report).

    Science.gov (United States)

    Tahara, Shoichi; Yamamoto, Sumiyo; Yamajima, Yukiko; Miyakawa, Hiroyuki; Uematsu, Yoko; Monma, Kimio

    2017-01-01

    Following the previous report, a rapid dialysis method was developed for the extraction and purification of four artificial sweeteners, namely, sodium saccharide (Sa), acesulfame potassium (AK), aspartame (APM), and dulcin (Du), which are present in various foods. The method was evaluated by the addition of 0.02 g/kg of these sweeteners to a cookie sample, in the same manner as in the previous report. Revisions from the previous method were: reduction of the total dialysis volume from 200 to 100 mL, change of tube length from 55 to 50 cm, change of dialysate from 0.01 mol/L hydrochloric aqueous solution containing 10% sodium chloride to 30% methanol solution, and change of dialysis conditions from ambient temperature with occasional shaking to 50℃ with shaking at 160 rpm. As a result of these revisions, the recovery reached 99.3-103.8% with one hour dialysis. The obtained recovery yields were comparable to the recovery yields in the previous method with four hour dialysis.

  8. ReagentTF: a rapid and versatile optical clearing method for biological imaging(Conference Presentation)

    Science.gov (United States)

    Yu, Tingting; Zhu, Jingtan; Li, Yusha; Qi, Yisong; Xu, Jianyi; Gong, Hui; Luo, Qingming; Zhu, Dan

    2017-02-01

    The emergence of various optical clearing methods provides a great potential for imaging deep inside tissues by combining with multiple-labelling and microscopic imaging techniques. They were generally developed for specific imaging demand thus presented some non-negligible limitations such as long incubation time, tissue deformation, fluorescence quenching, incompatibility with immunostaining or lipophilic tracers. In this study, we developed a rapid and versatile clearing method, termed ReagentTF, for deep imaging of various fluorescent samples. This method can not only efficiently clear embryos, neonatal whole-brains and adult thick brain sections by simple immersion in aqueous mixtures with minimal volume change, but also can preserve fluorescence of various fluorescent proteins and simultaneously be compatible with immunostaining and lipophilic neuronal dyes. We demonstrate the effectiveness of this method in reconstructing the cell distributions of mouse hippocampus, visualizing the neural projection from CA1 (Cornu Ammonis 1) to HDB (nucleus of the horizontal limb of the diagonal band), and observing the growth of forelimb plexus in whole-mount embryos. These results suggest that ReagentTF is useful for large-volume imaging and will be an option for the deep imaging of biological tissues.

  9. Computationally rapid method of estimating signal-to-noise ratio for phased array image reconstructions.

    Science.gov (United States)

    Wiens, Curtis N; Kisch, Shawn J; Willig-Onwuachi, Jacob D; McKenzie, Charles A

    2011-10-01

    Measuring signal-to-noise ratio (SNR) for parallel MRI reconstructions is difficult due to spatially dependent noise amplification. Existing approaches for measuring parallel MRI SNR are limited because they are not applicable to all reconstructions, require significant computation time, or rely on repeated image acquisitions. A new SNR estimation approach is proposed, a hybrid of the repeated image acquisitions method detailed in the National Electrical Manufacturers Association (NEMA) standard and the Monte Carlo based pseudo-multiple replica method, in which the difference between images reconstructed from the unaltered acquired data and that same data reconstructed after the addition of calibrated pseudo-noise is used to estimate the noise in the parallel MRI image reconstruction. This new noise estimation method can be used to rapidly compute the pixel-wise SNR of the image generated from any parallel MRI reconstruction of a single acquisition. SNR maps calculated with the new method are validated against existing SNR calculation techniques. Copyright © 2011 Wiley-Liss, Inc.

  10. A rapid and sensitive method for the detection of aromatic amines in cosmetics.

    Science.gov (United States)

    Hailong, Xiao; Fen, Qian; Ying, Xu; Jianhong, Pan; Haiyun, Tu; Hongqing, Wang; Saijun, Lin; Jichun, Han

    2014-02-01

    Aromatic amines (AAs) are common chemical pollutants and banned ingredients in cosmetics. In this study, a rapid, simple and stable method for the detection of nine AAs in cosmetics was established based on the optimization of cation exchange solid-phase extraction and liquid chromatography tandem mass spectrometry. The method displayed good linearity within a range of 2-1,000 µg/kg, with limits of quantitation at the level of µg/kg for cosmetic samples. The recoveries obtained for all analyzed amines ranged between 83.6 and 97.8%, and the repeatability (r) and reproducibility (R) values indicated that all nine AAs showed good precision (r ≤ 4.5% and R ≤ 7.7%). The method was applied for the detection of 36 cosmetic samples. It was found that the primary pollutants of AAs were 3, 3'-dichlorobenzidine and 4-aminoazobenzene. The total amine concentration in cosmetic samples ranged from 880 to 5,200 µg/kg. The proposed method is applicable for the analysis of most cosmetic samples.

  11. A simple and rapid method to characterize lipid fate in skeletal muscle.

    Science.gov (United States)

    Massart, Julie; Zierath, Juleen R; Chibalin, Alexander V

    2014-06-24

    Elevated fatty acids contribute to the development of type 2 diabetes and affect skeletal muscle insulin sensitivity. Since elevated intramuscular lipids and insulin resistance is strongly correlated, aberrant lipid storage or lipid intermediates may be involved in diabetes pathogenesis. The aim of this study was to develop a method to determine the dynamic metabolic fate of lipids in primary human skeletal muscle cells and in intact mouse skeletal muscle. We report a simple and fast method to characterize lipid profiles in skeletal muscle using thin layer chromatography. The described method was specifically developed to assess lipid utilization in cultured and intact skeletal muscle. We determined the effect of a pan-diacylglycerol kinase (DGK) class I inhibitor (R59949) on lipid metabolism to validate the method. In human skeletal muscle cells, DGK inhibition impaired diacylglycerol (DAG) conversion to phosphatidic acid and increased triglyceride synthesis. In intact glycolytic mouse skeletal muscle, DGK inhibition triggered the accumulation of DAG species. Conversely, the DGK inhibitor did not affect DAG content in oxidative muscle. This simple assay detects rapid changes in the lipid species composition of skeletal muscle with high sensitivity and specificity. Determination of lipid metabolism in skeletal muscle may further elucidate the mechanisms contributing to the pathogenesis of insulin resistance in type 2 diabetes or obesity.

  12. A new rapid method for Clostridium difficile DNA extraction and detection in stool: toward point-of-care diagnostic testing

    National Research Council Canada - National Science Library

    Freifeld, Alison G; Simonsen, Kari A; Booth, Christine S; Zhao, Xing; Whitney, Scott E; Karre, Teresa; Iwen, Peter C; Viljoen, Hendrik J

    2012-01-01

    We describe a new method for the rapid diagnosis of Clostridium difficile infection, with stool sample preparation and DNA extraction by heat and physical disruption in a single-use lysis microreactor (LMR...

  13. A novel method for ATLAS FSI alignment based on rapid, direct phase monitoring

    CERN Document Server

    Gibson, S M; The ATLAS collaboration; Horton, K; Lewis, A; Liang, Z; Livermore, S; Mattravers, C; Nickerson, R B

    2010-01-01

    Frequency Scanning Interferometry is a precise, multiple distance measurement technique, originally developed for ATLAS, which is suited to a variety of applications in the survey and alignment of future accelerators and particle detectors. The ATLAS inner detector is instrumented with an automated FSI alignment system, capable of simultaneously measuring hundreds of interferometers within the operational particle tracker. The alignment system began data taking in 2008 and we present the latest results from the on-detector system during LHC running. A new method has been developed based on rapid, direct monitoring of the interferometer phase, which allows the measurement of short term motions with improved precision, at a fraction of the wavelength of light (typically sensitive to < 50 nm). We outline the theory behind this novel technique and demonstrate precise measurements from ATLAS, which reveal interesting micron-level movements of the inner detector, correlated with thermal cycles and magnetic f...

  14. Apparatus and method for rapid cooling of large area substrates in vacuum

    Science.gov (United States)

    Barth, Kurt L.; Enzenroth, Robert A.; Sampath, Walajabad S.

    2010-09-28

    The present invention is directed to an apparatus and method for rapid cooling of a large substrate in a vacuum environment. A first cooled plate is brought into close proximity with one surface of a flat substrate. The spatial volume between the first cooling plate and the substrate is sealed and brought to a higher pressure than the surrounding vacuum level to increase the cooling efficiency. A second cooled plate is brought into close proximity with the opposite surface of the flat substrate. A second spatial volume between the second cooling plate and the substrate is sealed and the gas pressure is equalized to the gas pressure in the first spatial volume. The equalization of the gas pressure on both sides of the flat substrate eliminates deflection of the substrate and bending stress in the substrate.

  15. A rapid and economic in-house DNA purification method using glass syringe filters.

    Directory of Open Access Journals (Sweden)

    Yun-Cheol Kim

    Full Text Available BACKGROUND: Purity, yield, speed and cost are important considerations in plasmid purification, but it is difficult to achieve all of these at the same time. Currently, there are many protocols and kits for DNA purification, however none maximize all four considerations. METHODOLOGY/PRINCIPAL FINDINGS: We now describe a fast, efficient and economic in-house protocol for plasmid preparation using glass syringe filters. Plasmid yield and quality as determined by enzyme digestion and transfection efficiency were equivalent to the expensive commercial kits. Importantly, the time required for purification was much less than that required using a commercial kit. CONCLUSIONS/SIGNIFICANCE: This method provides DNA yield and quality similar to that obtained with commercial kits, but is more rapid and less costly.

  16. A robust and rapid method of producing soluble, stable, and functional G-protein coupled receptors.

    Directory of Open Access Journals (Sweden)

    Karolina Corin

    Full Text Available Membrane proteins, particularly G-protein coupled receptors (GPCRs, are notoriously difficult to express. Using commercial E. coli cell-free systems with the detergent Brij-35, we could rapidly produce milligram quantities of 13 unique GPCRs. Immunoaffinity purification yielded receptors at >90% purity. Secondary structure analysis using circular dichroism indicated that the purified receptors were properly folded. Microscale thermophoresis, a novel label-free and surface-free detection technique that uses thermal gradients, showed that these receptors bound their ligands. The secondary structure and ligand-binding results from cell-free produced proteins were comparable to those expressed and purified from HEK293 cells. Our study demonstrates that cell-free protein production using commercially available kits and optimal detergents is a robust technology that can be used to produce sufficient GPCRs for biochemical, structural, and functional analyses. This robust and simple method may further stimulate others to study the structure and function of membrane proteins.

  17. Rapid method for surveying CO concentrations in high-rise buildings

    Energy Technology Data Exchange (ETDEWEB)

    Flachsbart, P.G.; Ott, W.R.

    1986-01-01

    A rapid method for employing personal exposure monitors (PEMs) to measure carbon monoxide (CO) concentrations in high-rise buildings is described. The purpose is to determine whether or not a CO problem exists in a building, and, if so, what corrective actions should be taken. The methodology was applied to a 15-story building in Palo Alto, CA, where elevated CO concentrations were discovered on the first 11 floors. The source appeared to be an underground parking garage. A follow-up survey four years later revealed that mitigative measures designed to reduce these concentrations had been successful. The survey methodology is inexpensive and can be applied to a number of buildings in a city.

  18. Evaluation of rapid alternative methods for drug susceptibility testing in clinical isolates of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Luciano Mengatto

    2006-08-01

    Full Text Available A study was carried out to compare the performance of a commercial method (MGIT and four inexpensive drug susceptibility methods: nitrate reductase assay (NRA, microscopic observation drug susceptibility (MODS assay, MTT test, and broth microdilution method (BMM. A total of 64 clinical isolates of Mycobacterium tuberculosis were studied. The Lowenstein-Jensen proportion method (PM was used as gold standard. MGIT, NRA, MODS, and MTT results were available on an average of less than 10 days, whereas BMM results could be reported in about 20 days. Most of the evaluated tests showed excellent performance for isoniazid and rifampicin, with sensitivity and specificity values > 90%. With most of the assays, sensitivity for ethambutol was low (62-87% whereas for streptomycin, sensitivity values ranged from 84 to 100%; NRA-discrepancies were associated with cultures with a low proportion of EMB-resistant organisms while most discrepancies with quantitative tests (MMT and BMM were seen with isolates whose minimal inhibitory concentrations fell close the cutoff. MGIT is reliable but still expensive. NRA is the most inexpensive and easiest method to perform without changing the organization of the routine PM laboratory performance. While MODS, MTT, and BMM, have the disadvantage from the point of view of biosafety, they offer the possibility of detecting partial resistant strains. This study shows a very good level of agreement of the four low-cost methods compared to the PM for rapid detection of isoniazid, rifampicin and streptomycin resistance (Kappa values > 0.8; more standardization is needed for ethambutol.

  19. A rapid and efficient method for assessing pathogenicity of ustilago maydis on maize and teosinte lines.

    Science.gov (United States)

    Chavan, Suchitra; Smith, Shavannor M

    2014-01-03

    Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is a biotrophic fungal pathogen and the causal agent of corn smut on maize. This disease is responsible for significant yield losses of approximately $1.0 billion annually in the U.S.(1) Several methods including crop rotation, fungicide application and seed treatments are currently used to control corn smut(2). However, host resistance is the only practical method for managing corn smut. Identification of crop plants including maize, wheat, and rice that are resistant to various biotrophic pathogens has significantly decreased yield losses annually(3-5). Therefore, the use of a pathogen inoculation method that efficiently and reproducibly delivers the pathogen in between the plant leaves, would facilitate the rapid identification of maize lines that are resistant to U. maydis. As, a first step toward indentifying maize lines that are resistant to U. maydis, a needle injection inoculation method and a resistance reaction screening method was utilized to inoculate maize, teosinte, and maize x teosinte introgression lines with a U. maydis strain and to select resistant plants. Maize, teosinte and maize x teosinte introgression lines, consisting of about 700 plants, were planted, inoculated with a strain of U. maydis, and screened for resistance. The inoculation and screening methods successfully identified three teosinte lines resistant to U. maydis. Here a detailed needle injection inoculation and resistance reaction screening protocol for maize, teosinte, and maize x teosinte introgression lines is presented. This study demonstrates that needle injection inoculation is an invaluable tool in agriculture that can efficiently deliver U. maydis in between the plant leaves and has provided plant lines that are resistant to U. maydis that can now be combined and tested in breeding programs for improved disease resistance.

  20. A method for rapid measurement of laser ablation rate of hard dental tissue

    Science.gov (United States)

    Perhavec, T.; Gorkič, A.; Bračun, D.; Diaci, J.

    2009-06-01

    The aim of the study reported here is the development of a new method which allows rapid and accurate in-vitro measurements of three-dimensional (3D) shape of laser ablated craters in hard dental tissues and the determination of crater volume, ablation rate and speed. The method is based on the optical triangulation principle. A laser sheet projector illuminates the surface of a tooth, mounted on a linear translation stage. As the tooth is moved by the translation stage a fast digital video camera captures series of images of the illuminated surface. The images are analyzed to determine a 3D model of the surface. Custom software is employed to analyze the 3D model and to determine the volume of the ablated craters. Key characteristics of the method are discussed as well as some practical aspects pertinent to its use. The method has been employed in an in-vitro study to examine the ablation rates and speeds of the two main laser types currently employed in dentistry, Er:YAG and Er,Cr:YSGG. Ten samples of extracted human molar teeth were irradiated with laser pulse energies from 80 mJ to the maximum available energy (970 mJ with the Er:YAG, and 260 mJ with the Er,Cr:YSGG). About 2000 images of each ablated tooth surface have been acquired along a translation range of 10 mm, taking about 10 s and providing close to 1 million surface measurement points. Volumes of 170 ablated craters (half of them in dentine and the other half in enamel) were determined from this data and used to examine the ablated volume per pulse energy and ablation speed. The results show that, under the same conditions, the ablated volume per pulse energy achieved by the Er:YAG laser exceeds that of the Er,Cr:YSGG laser in almost all regimes for dentine and enamel. The maximum Er:YAG laser ablation speeds (1.2 mm 3/s in dentine and 0.7 mm 3/s in enamel) exceed those obtained by the Er,Cr:YSGG laser (0.39 mm 3/s in dentine and 0.12 mm 3/s in enamel). Since the presented method proves to be easy to

  1. Rapid, convenient method for screening imidazole-containing compounds for heme oxygenase inhibition.

    Science.gov (United States)

    Vlahakis, Jason Z; Rahman, Mona N; Roman, Gheorghe; Jia, Zongchao; Nakatsu, Kanji; Szarek, Walter A

    2011-01-01

    Sensitive assays for measuring heme oxygenase activity have been based on the gas-chromatographic detection of carbon monoxide using elaborate, expensive equipment. The present study describes a rapid and convenient method for screening imidazole-containing candidates for inhibitory activity against heme oxygenase using a plate reader, based on the spectroscopic evaluation of heme degradation. A PowerWave XS plate reader was used to monitor the absorbance (as a function of time) of heme bound to purified truncated human heme oxygenase-1 (hHO-1) in the individual wells of a standard 96-well plate (with or without the addition of a test compound). The degradation of heme by heme oxygenase-1 was initiated using l-ascorbic acid, and the collected relevant absorbance data were analyzed by three different methods to calculate the percent control activity occurring in wells containing test compounds relative to that occurring in control wells with no test compound present. In the cases of wells containing inhibitory compounds, significant shifts in λ(max) from 404 to near 412 nm were observed as well as a decrease in the rate of heme degradation relative to that of the control. Each of the three methods of data processing (overall percent drop in absorbance over 1.5h, initial rate of reaction determined over the first 5 min, and estimated pseudo first-order reaction rate constant determined over 1.5h) gave similar and reproducible results for percent control activity. The fastest and easiest method of data analysis was determined to be that using initial rates, involving data acquisition for only 5 min once reactions have been initiated using l-ascorbic acid. The results of the study demonstrate that this simple assay based on the spectroscopic detection of heme represents a rapid, convenient method to determine the relative inhibitory activity of candidate compounds, and is useful in quickly screening a series or library of compounds for heme oxygenase inhibition

  2. A rapid method of fruit cell isolation for cell size and shape measurements

    Directory of Open Access Journals (Sweden)

    Johnston Jason W

    2009-04-01

    Full Text Available Abstract Background Cell size is a structural component of fleshy fruit, contributing to important traits such as fruit size and texture. There are currently a number of methods for measuring cell size; most rely either on tissue sectioning or digestion of the tissue with cell wall degrading enzymes or chemicals to release single cells. Neither of these approaches is ideal for assaying large fruit numbers as both require a considerable time to prepare the tissue, with current methods of cell wall digestions taking 24 to 48 hours. Additionally, sectioning can lead to a measurement of a plane that does not represent the widest point of the cell. Results To develop a more rapid way of measuring fruit cell size we have developed a protocol that solubilises pectin in the middle lamella of the plant cell wall releasing single cells into a buffered solution. Gently boiling small fruit samples in a 0.05 M Na2CO3 solution, osmotically balanced with 0.3 M mannitol, produced good cell separation with little cellular damage in less than 30 minutes. The advantage of combining a chemical treatment with boiling is that the cells are rapidly killed. This stopped cell shape changes that could potentially occur during separation. With this method both the rounded and angular cells of the apple cultivars SciRos 'Pacific Rose' and SciFresh 'Jazz'™ were observed in the separated cells. Using this technique, an in-depth analysis was performed measuring cell size from 5 different apple cultivars. Cell size was measured using the public domain ImageJ software. For each cultivar a minimum of 1000 cells were measured and it was found that each cultivar displayed a different distribution of cell size. Cell size within cultivars was similar and there was no correlation between flesh firmness and cell size. This protocol was tested on tissue from other fleshy fruit including tomato, rock melon and kiwifruit. It was found that good cell separation was achieved with flesh

  3. Use of predictive models and rapid methods to nowcast bacteria levels at coastal beaches

    Science.gov (United States)

    Francy, Donna S.

    2009-01-01

    The need for rapid assessments of recreational water quality to better protect public health is well accepted throughout the research and regulatory communities. Rapid analytical methods, such as quantitative polymerase chain reaction (qPCR) and immunomagnetic separation/adenosine triphosphate (ATP) analysis, are being tested but are not yet ready for widespread use.Another solution is the use of predictive models, wherein variable(s) that are easily and quickly measured are surrogates for concentrations of fecal-indicator bacteria. Rainfall-based alerts, the simplest type of model, have been used by several communities for a number of years. Deterministic models use mathematical representations of the processes that affect bacteria concentrations; this type of model is being used for beach-closure decisions at one location in the USA. Multivariable statistical models are being developed and tested in many areas of the USA; however, they are only used in three areas of the Great Lakes to aid in notifications of beach advisories or closings. These “operational” statistical models can result in more accurate assessments of recreational water quality than use of the previous day's Escherichia coli (E. coli)concentration as determined by traditional culture methods. The Ohio Nowcast, at Huntington Beach, Bay Village, Ohio, is described in this paper as an example of an operational statistical model. Because predictive modeling is a dynamic process, water-resource managers continue to collect additional data to improve the predictive ability of the nowcast and expand the nowcast to other Ohio beaches and a recreational river. Although predictive models have been shown to work well at some beaches and are becoming more widely accepted, implementation in many areas is limited by funding, lack of coordinated technical leadership, and lack of supporting epidemiological data.

  4. Rapid radiometric methods to detect and differentiate Mycobacterium tuberculosis/M. bovis from other mycobacterial species

    Energy Technology Data Exchange (ETDEWEB)

    Siddiqi, S.H.; Hwangbo, C.C.; Silcox, V.; Good, R.C.; Snider, D.E. Jr.; Middlebrook, G.

    1984-10-01

    Rapid methods for the differentiation of Mycobacterium tuberculosis/M. bovis (TB complex) from other mycobacteria (MOTT bacilli) were developed and evaluated in a three-phase study. In the first phase, techniques for identification of Mycobacterium species were developed by using radiometric technology and BACTEC Middlebrook 7H12 liquid medium. Based on /sup 14/CO/sub 2/ evolution, characteristic growth patterns were established for 13 commonly encountered mycobacterial species. Mycobacteria belonging to the TB complex were differentiated from other mycobacteria by cellular morphology and rate of /sup 14/CO/sub 2/ evolution. For further differentiation, radiometric tests for niacin production and inhibition by Q-nitro-alpha-acetyl amino-beta-hydroxy-propiophenone (NAP) were developed. In the second phase, 100 coded specimens on Lowenstein-Jensen medium were identified as members of the TB complex, MOTT bacilli, bacteria other than mycobacteria, or ''no viable organisms'' within 3 to 12 (average 6.4) days of receipt from the Centers for Disease Control. Isolation and identification of mycobacteria from 20 simulated sputum specimens were carried out in phase III. Out of 20 sputum specimens, 16 contained culturable mycobacteria, and all of the positives were detected by the BACTEC method in an average of 7.3 days. The positive mycobacterial cultures were isolated and identified as TB complex or MOTT bacilli in an average of 12.8 days. The radiometric NAP test was found to be highly sensitive and specific for a rapid identification of TB complex, whereas the radiometric niacin test was found to have some inherent problems. Radiometric BACTEC and conventional methodologies were in complete agreement in Phase II as well as in Phase III.

  5. A rapid minor groove binder PCR method for distinguishing the vaccine strain Brucella abortus 104M.

    Science.gov (United States)

    Nan, Wenlong; Qin, Lide; Wang, Yong; Zhang, Yueyong; Tan, Pengfei; Chen, Yuqi; Mao, Kairong; Chen, Yiping

    2018-01-24

    Brucellosis is a widespread zoonotic disease caused by Gram-negative Brucella bacteria. Immunisation with attenuated vaccine is an effective method of prevention, but it can interfere with diagnosis. Live, attenuated Brucella abortus strain 104M has been used for the prevention of human brucellosis in China since 1965. However, at present, no fast and reliable method exists that can distinguish this strain from field strains. Single nucleotide polymorphism (SNP)-based assays offer a new approach for such discrimination. SNP-based minor groove binder (MGB) and Cycleave assays have been used for rapid identification of four Brucella vaccine strains (B. abortus strains S19, A19 and RB51, and B. melitensis Rev1). The main objective of this study was to develop a PCR assay for rapid and specific detection of strain 104M. We developed a SNP-based MGB PCR assay that could successfully distinguish strain 104M from 18 representative strains of Brucella (B. abortus biovars 1, 2, 3, 4, 5, 6, 7 and 9, B. melitensis biovars 1, 2 and 3, B. suis biovars 1, 2, 3 and 4, B. canis, B. neotomae, and B. ovis), four Brucella vaccine strains (A19, S19, S2, M5), and 55 Brucella clinical field strains. The assay gave a negative reaction with four non-Brucella species (Escherichia coli, Pasteurella multocida, Streptococcus suis and Pseudomonas aeruginosa). The minimum sensitivity of the assay, evaluated using 10-fold dilutions of chromosomal DNA, was 220 fg for the 104M strain and 76 fg for the single non-104M Brucella strain tested (B. abortus A19). The assay was also reproducible (intra- and inter-assay coefficients of variation = 0.006-0.022 and 0.012-0.044, respectively). A SNP-based MGB PCR assay was developed that could straightforwardly and unambiguously distinguish B. abortus vaccine strain 104M from non-104M Brucella strains. Compared to the classical isolation and identification approaches of bacteriology, this real-time PCR assay has substantial advantages in terms of

  6. A Method for Rapid Measurement of Contrast Sensitivity on Mobile Touch-Screens

    Science.gov (United States)

    Mulligan, Jeffrey B.

    2016-01-01

    Touch-screen displays in cell phones and tablet computers are now pervasive, making them an attractive option for vision testing outside of the laboratory or clinic. Here we de- scribe a novel method in which subjects use a finger swipe to indicate the transition from visible to invisible on a grating which is swept in both contrast and frequency. Because a single image can be swiped in about a second, it is practical to use a series of images to zoom in on particular ranges of contrast or frequency, both to increase the accuracy of the measurements and to obtain an estimate of the reliability of the subject. Sensitivities to chromatic and spatio-temporal modulations are easily measured using the same method. A proto- type has been developed for Apple Computer's iPad/iPod/iPhone family of devices, implemented using an open-source scripting environment known as QuIP (QUick Image Processing, http://hsi.arc.nasa.gov/groups/scanpath/research.php). Preliminary data show good agreement with estimates obtained from traditional psychophysical methods as well as newer rapid estimation techniques. Issues relating to device calibration are also discussed.

  7. Modified agar dilution method for rapid antibiotic susceptibility testing of anaerobic bacteria.

    Science.gov (United States)

    Hanson, C W; Martin, W J

    1978-01-01

    A simplified method has been developed for agar dilution antimicrobial susceptibility testing of anaerobic bacteria, designed to economize on time and money when only a few isolates need to be tested. The procedure is based on the principle of using filter paper disks as carriers of the antibiotic and 35- by 10-mm petri dishes which, when inoculated with the Steers replicator, can test up to four organisms per plate. The procedure was run in parallel with conventional agar dilution techniques and showed 95% agreement to within one dilution for all minimal inhibitory concentrations recorded on fresh anaerobic isolates from clinical specimens. The technique was further simplified by using commercially available antibiotic-containing disks, thereby alleviating the tedious and time-consuming procedure of preparing the disks. The data indicated that 48- to 72-h diffusion periods were sufficient to achieve a uniform concentration of the antibiotic in the petri plate and that the antibiotics were stable at room temperature for that period of time. In terms of applicability and relevance to the needs of the clinical microbiology laboratory, the modified agar dilution method for rapid antimicrobial susceptibility testing of individual anaerobic isolates was found to be superior to the broth dilution method since it was easier to read and required considerably less set up time. PMID:400819

  8. Development of a rapid HRM genotyping method for detection of dog-derived Giardia lamblia.

    Science.gov (United States)

    Tan, Liping; Yu, Xingang; Abdullahi, Auwalu Yusuf; Wu, Sheng; Zheng, Guochao; Hu, Wei; Song, Meiran; Wang, Zhen; Jiang, Biao; Li, Guoqing

    2015-11-01

    Giardia lamblia is a zoonotic flagellate protozoan in the intestine of human and many mammals including dogs. To assess a threat of dog-derived G. lamblia to humans, the common dog-derived G. lamblia assemblages A, C, and D were genotyped by high-resolution melting (HRM) technology. According to β-giardin gene sequence, the qPCR-HRM primers BG5 and BG7 were designed. A series of experiments on the stability, sensitivity, and accuracy of the HRM method were also tested. Results showed that the primers BG5 and BG7 could distinguish among three assemblages A, C, and D, which Tm value differences were about 1 °C to each other. The melting curves of intra-assay reproducibility were almost coincided, and those of inter-assay reproducibility were much the same shape. The lowest detection concentration was about 5 × 10(-6)-ng/μL sample. The genotyping results from 21 G. lamblia samples by the HRM method were in complete accordance with sequencing results. It is concluded that the HRM genotyping method is rapid, stable, specific, highly sensitive, and suitable for clinical detection and molecular epidemiological survey of dog-derived G. lamblia.

  9. Rapid, high-temperature, field test method for evaluation of geothermal calcium carbonate scale inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Asperger, R.G.

    1986-09-01

    A new test method is described that allows the rapid field testing of calcium carbonate scale inhibitors at 500/sup 0/F (260/sup 0/C). The method evolved from use of a full-flow test loop on a well with a mass flow rate of about 1 x 10/sup 6/ lbm/hr (126 kg/s). It is a simple, effective way to evaluate the effectiveness of inhibitors under field conditions. Five commercial formulations were chosen for field evaluation on the basis of nonflowing, laboratory screening tests at 500/sup 0/F (260/sup 0/C). Four of these formulations from different suppliers controlled calcium carbonate scale deposition as measured by the test method. Two of these could dislodge recently deposited scale that had not age-hardened. Performance-profile diagrams, which were measured for these four effective inhibitors, show the concentration interrelationship between brine calcium and inhibitor concentrations at which the formulations will and will not stop scale formation in the test apparatus. With these diagrams, one formulation was chosen for testing on the full-flow brine line. The composition was tested for 6 weeks and showed a dramatic decrease in the scaling occurring at the flow-control valve. This scaling was about to force a shutdown of a major, long-term flow test being done for reservoir economic evaluations. The inhibitor stopped the scaling, and the test was performed without interruption.

  10. A simple and rapid cultural method for detection of Enterobacter sakazakii in environmental samples.

    Science.gov (United States)

    Guillaume-Gentil, O; Sonnard, V; Kandhai, M C; Marugg, J D; Joosten, H

    2005-01-01

    A method was developed to detect and identify Enterobacter sakazakii in environmental samples. The method is based on selective enrichment at 45+/-0.5 degrees C in lauryl sulfate tryptose broth supplemented with 0.5 M NaCl and 10 mg/liter vancomycin (mLST) for 22 to 24 h followed by streaking on tryptone soy agar with bile salts. When exposed to light during incubation at 37 degrees C, E. sakazakii produces yellow colonies within 24 h; identification was confirmed by testing for alpha-glucosidase activity and by using API 20E strips. All of the E. sakazakii strains tested (n = 99) were able to grow in mLST at 45+/-0.5 degrees C, whereas 35 of 39 strains of potential competitors, all belonging to the Enterobacteriaceae, were suppressed. A survey was carried out with 192 environmental samples from four different milk powder factories. Using this new protocol, E. sakazakii was isolated from almost 40% of the samples, whereas the reference procedure (enrichment in buffered peptone water, isolation on violet red bile glucose agar, and biochemical identification of randomly chosen colonies) only yielded 26% positive results. This selective method can be very useful for the rapid and reliable detection of E. sakazakii in environmental samples.

  11. Rapid, Simple, and Sensitive Spectrofluorimetric Method for the Estimation of Ganciclovir in Bulk and Pharmaceutical Formulations

    Directory of Open Access Journals (Sweden)

    Garima Balwani

    2013-01-01

    Full Text Available A new, simple, rapid, sensitive, accurate, and affordable spectrofluorimetric method was developed and validated for the estimation of ganciclovir in bulk as well as in marketed formulations. The method was based on measuring the native fluorescence of ganciclovir in 0.2 M hydrochloric acid buffer of pH 1.2 at 374 nm after excitation at 257 nm. The calibration graph was found to be rectilinear in the concentration range of 0.25–2.00 μg mL−1. The limit of quantification and limit of detection were found to be 0.029 μg mL−1 and 0.010 μg mL−1, respectively. The method was fully validated for various parameters according to ICH guidelines. The results demonstrated that the procedure is accurate, precise, and reproducible (relative standard deviation <2% and can be successfully applied for the determination of ganciclovir in its commercial capsules with average percentage recovery of 101.31 ± 0.90.

  12. Rapid Method for the Determination of the Stable Oxygen Isotope Ratio of Water in Alcoholic Beverages.

    Science.gov (United States)

    Wang, Daobing; Zhong, Qiding; Li, Guohui; Huang, Zhanbin

    2015-10-28

    This paper demonstrates the first successful application of an online pyrolysis technique for the direct determination of oxygen isotope ratios (δ(18)O) of water in alcoholic beverages. Similar water concentrations in each sample were achieved by adjustment with absolute ethyl alcohol, and then a fixed GC split ratio can be used. All of the organic ingredients were successfully separated from the analyte on a CP-PoraBond Q column and subsequently vented out, whereas water molecules were transferred into the reaction furnace and converted to CO. With the system presented, 15-30 μL of raw sample was diluted and can be analyzed repeatedly; the analytical precision was better than 0.4‰ (n = 5) in all cases, and more than 50 injections can be made per day. No apparent memory effect was observed even if water samples were injected using the same syringe; a strong correlation (R(2) = 0.9998) was found between the water δ(18)O of measured sample and that of working standards. There was no significant difference (p > 0.05) between the mean δ(18)O value and that obtained by the traditional method (CO2-water equilibration/isotope ratio mass spectrometry) and the newly developed method in this study. The advantages of this new method are its rapidity and straightforwardness, and less test portion is required.

  13. A rapid and efficient electroporation method for transformation of Halomonas sp. O-1.

    Science.gov (United States)

    Harris, Joshua R; Lundgren, Benjamin R; Grzeskowiak, Brian R; Mizuno, Kouhei; Nomura, Christopher T

    2016-10-01

    Halomonas sp. O-1 is a halophilic bacterium with a high potential for industrial application due to its natural ability to produce polyhydroxyalkanoates (PHAs) using seawater-based media. However, a major barrier preventing industrial scale implementation of this organism is a lack of molecular methodologies capable of readily transforming members of the Halomonas genus. Currently, the only reliable method used for introducing DNA into Halomonas spp. is bacterial conjugation, a somewhat tedious and time-consuming technique compared to electroporation-based methodologies. Here we describe a rapid and reproducible method for the electroporation of Halomonas sp. O-1 with plasmid DNA. Electrocompetent cells were generated by growing Halomonas sp. O-1 in a yeast extract-tryptone medium with a final salinity of 3.5%, pH of 7.5, followed by several washes using 300mM sucrose. Results show that plasmids containing chloramphenicol (Cm(R)) and gentamicin (Gm(R)) resistance cassettes are suitable antibiotic selection markers for transformation and yields of 10(4) transformants per μg of DNA were obtained. This method is simple to perform and the materials used are readily available in most research labs. Additionally, this plasmid-based transformation procedure has the potential to be adapted for a number of applications including the creation of recombinant stains and the generation of deletion mutants of Halomonas spp. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. A Rapid and Sensitive Method to Measure the Functional Activity of Shiga Toxins in Human Serum

    Directory of Open Access Journals (Sweden)

    Valentina Arfilli

    2015-11-01

    Full Text Available Shiga toxins (Stx have a definite role in the development of hemolytic uremic syndrome in children with hemorrhagic colitis caused by pathogenic Stx-producing Escherichia coli (STEC strains. The dramatic effects of these toxins on the microvasculature of different organs, particularly of the kidney, are well known, whereas there is no consensus on the mechanism by which Stx reach the endothelia of target organs and/or indirectly injure these body sites. We hereby describe a quick (4 h, radioactive, Raji cell-based method designed for the detection of Stx in human sera. The assay monitors the translation impairment induced by these powerful inhibitors of protein synthesis, which are identified properly by neutralizing their activity with specific monoclonal antibodies. By this method, we detected for the first time the functional activity of Stx in sera of STEC-infected patients during hemorrhagic colitis. Recent research has pointed to a dynamic process of Stx-induced renal intoxication in which concurrent and interactive steps are involved. Our rapid and specific method could be useful for studying the kinetics of Stx during the natural course of STEC infection and the interplay between Stx activity in serum and Stx presence in different blood fractions (neutrophils, monocytes, platelets, leukocyte-platelet aggregates, microvesicles, lipoproteins.

  15. FLASH: A rapid method for prototyping paper-based microfluidic devices‡

    Science.gov (United States)

    Martinez, Andres W.; Phillips, Scott T.; Wiley, Benjamin J.; Gupta, Malancha

    2011-01-01

    This article describes FLASH (Fast Lithographic Activation of Sheets), a rapid method for laboratory prototyping of microfluidic devices in paper. Paper-based microfluidic devices are emerging as a new technology for applications in diagnostics for the developing world, where low cost and simplicity are essential. FLASH is based on photolithography, but requires only a UV lamp and a hotplate; no clean-room or special facilities are required (FLASH patterning can even be performed in sunlight if a UV lamp and hotplate are unavailable). The method provides channels in paper with dimensions as small as 200 μm in width and 70 μm in height; the height is defined by the thickness of the paper. Photomasks for patterning paper-based microfluidic devices can be printed using an ink-jet printer or photocopier, or drawn by hand using a waterproof black pen. FLASH provides a straightforward method for prototyping paper-based microfluidic devices in regions where the technological support for conventional photolithography is not available. PMID:19023478

  16. AO–MW–PLS method applied to rapid quantification of teicoplanin with near-infrared spectroscopy

    Directory of Open Access Journals (Sweden)

    Jiemei Chen

    2017-01-01

    Full Text Available Teicoplanin (TCP is an important lipoglycopeptide antibiotic produced by fermenting Actinoplanes teichomyceticus. The change in TCP concentration is important to measure in the fermentation process. In this study, a reagent-free and rapid quantification method for TCP in the TCP–Tris–HCl mixture samples was developed using near-infrared (NIR spectroscopy by focusing our attention on the fermentation process for TCP. The absorbance optimization (AO partial least squares (PLS was proposed and integrated with the moving window (MW PLS, which is called AO–MW–PLS method, to select appropriate wavebands. A model set that includes various wavebands that were equivalent to the optimal AO–MW–PLS waveband was proposed based on statistical considerations. The public region of all equivalent wavebands was just one of the equivalent wavebands. The obtained public regions were 1540–1868nm for TCP and 1114–1310nm for Tris. The root-mean-square error and correlation coefficient for leave-one-out cross validation were 0.046mg mL−1 and 0.9998mg mL−1 for TCP, and 0.235mg mL−1 and 0.9986mg mL−1 for Tris, respectively. All the models achieved highly accurate prediction effects, and the selected wavebands provided valuable references for designing specialized spectrometers. This study provided a valuable reference for further application of the proposed methods to TCP fermentation broth and to other spectroscopic analysis fields.

  17. A simple electrochemical method for the rapid estimation of antioxidant potentials of some selected medicinal plants.

    Science.gov (United States)

    Amidi, Salimeh; Mojab, Faraz; Bayandori Moghaddam, Abdolmajid; Tabib, Kimia; Kobarfard, Farzad

    2012-01-01

    Clinical and Epidemiological studies have shown that a diet rich in fruits and vegetables is associated with a decreased risk of cardiovascular diseases, cancers and other related disorders. These beneficial health effects have been attributed in part to the presence of antioxidants in dietary plants. Therefore screening for antioxidant properties of plant extracts has been one of the interests of scientists in this field. Different screening methods have been reported for the evaluation of antioxidant properties of plant extracts in the literature. In the present research a rapid screening method has been introduced based on cyclic voltammetry for antioxidant screening of some selected medicinal plant extracts. CYCLIC VOLTAMMETRY OF METHANOLIC EXTRACTS OF SEVEN MEDICINAL PLANTS: Buxus hyrcana, Rumex crispus, Achillea millefolium, Zataria multiflora, Ginkgo biloba, Lippia citriodora and Heptaptera anisoptera was carried out at different scan rates. Based on the interpretation of voltammograms, Rumex crispus, Achillea millefolium and Ginkgo biloba showed higher antioxidant capability than the others while Lippia citriodora contained the highest amount of antioxidants. Cyclic voltammetry is expected to be a simple method for screening antioxidants and estimating the antioxidant activity of foods and medicinal plants.

  18. A Rapid and Sensitive Method to Measure the Functional Activity of Shiga Toxins in Human Serum

    Science.gov (United States)

    Arfilli, Valentina; Carnicelli, Domenica; Ardissino, Gianluigi; Torresani, Erminio; Scavia, Gaia; Brigotti, Maurizio

    2015-01-01

    Shiga toxins (Stx) have a definite role in the development of hemolytic uremic syndrome in children with hemorrhagic colitis caused by pathogenic Stx-producing Escherichia coli (STEC) strains. The dramatic effects of these toxins on the microvasculature of different organs, particularly of the kidney, are well known, whereas there is no consensus on the mechanism by which Stx reach the endothelia of target organs and/or indirectly injure these body sites. We hereby describe a quick (4 h), radioactive, Raji cell-based method designed for the detection of Stx in human sera. The assay monitors the translation impairment induced by these powerful inhibitors of protein synthesis, which are identified properly by neutralizing their activity with specific monoclonal antibodies. By this method, we detected for the first time the functional activity of Stx in sera of STEC-infected patients during hemorrhagic colitis. Recent research has pointed to a dynamic process of Stx-induced renal intoxication in which concurrent and interactive steps are involved. Our rapid and specific method could be useful for studying the kinetics of Stx during the natural course of STEC infection and the interplay between Stx activity in serum and Stx presence in different blood fractions (neutrophils, monocytes, platelets, leukocyte-platelet aggregates, microvesicles, lipoproteins). PMID:26556372

  19. Rapid and sensitive method for determination of withaferin-A in human plasma by HPLC.

    Science.gov (United States)

    Patial, Pankaj; Gota, Vikram

    2011-02-01

    To develop and validate a rapid and sensitive high-performance liquid chromatographic method for determination of withaferin-A in human plasma. Withaferin-A, the active molecule of a traditional Indian herb, has demonstrated several biological activities in preclinical models. A validated bioassay is not available for its pharmacokinetic evaluation. The chromatographic system used a reverse-phase C18 column with UV-visible detection at 225 nm. The mobile phase consisted of water and acetonitrile applied in a gradient flow. Withaferin-A was extracted by simple protein-precipitation technique. The calibration curve was linear in the concentration range of 0.05-1.6 µg/ml. The method has the desired sensitivity to detect the plasma concentration range of withaferin-A that is likely to show biological activity based on in vitro data. This is the first HPLC method ever described for the estimation of withaferin-A in human plasma which could be applied for pharmacokinetic studies.

  20. Validation of a user-friendly and rapid method for quantifying iodine content of salt.

    Science.gov (United States)

    Rohner, Fabian; Garrett, Greg S; Laillou, Arnaud; Frey, Simone K; Mothes, Ralf; Schweigert, Florian J; Locatelli-Rossi, Lorenzo

    2012-12-01

    Despite considerable progress made in the past decade through salt iodization programs, over 2 billion people worldwide still have inadequate iodine intake, with devastating consequences for brain development and intellectual capacity. To optimize these programs with regard to salt iodine content, careful monitoring of salt iodine content is essential, but few methods are available to quantitatively measure iodine concentration in a simple, fast, and safe way. We have validated a newly developed device that quantitatively measures the content of potassium iodate in salt in a simple, safe, and rapid way. The linearity, determination and detection limit, and inter- and intra-assay variability of this colorimetric method were assessed and the method was compared with iodometric titration, using salt samples from several countries. Linearity of analysis ranged from 5 to 75 mg/kg iodine, with 1 mg/kg being the determination limit; the intra- and interassay imprecision was 0.9%, 0.5%, and 0.7% and 1.5%, 1.7%, and 2.5% for salt samples with iodine contents of 17, 30, and 55 mg/kg, respectively; the interoperator imprecision for the same samples was 1.2%, 4.9%, and 4.7%, respectively. Comparison with the iodometric method showed high agreement between the methods (R2 = 0.978; limits of agreement, -10.5 to 10.0 mg/kg). The device offers a field- and user-friendly solution to quantifying potassium iodate salt content reliably. For countries that use potassium iodide in salt iodization programs, further validation is required.

  1. Validity and feasibility of a satellite imagery-based method for rapid estimation of displaced populations

    Directory of Open Access Journals (Sweden)

    Checchi Francesco

    2013-01-01

    Full Text Available Abstract Background Estimating the size of forcibly displaced populations is key to documenting their plight and allocating sufficient resources to their assistance, but is often not done, particularly during the acute phase of displacement, due to methodological challenges and inaccessibility. In this study, we explored the potential use of very high resolution satellite imagery to remotely estimate forcibly displaced populations. Methods Our method consisted of multiplying (i manual counts of assumed residential structures on a satellite image and (ii estimates of the mean number of people per structure (structure occupancy obtained from publicly available reports. We computed population estimates for 11 sites in Bangladesh, Chad, Democratic Republic of Congo, Ethiopia, Haiti, Kenya and Mozambique (six refugee camps, three internally displaced persons’ camps and two urban neighbourhoods with a mixture of residents and displaced ranging in population from 1,969 to 90,547, and compared these to “gold standard” reference population figures from census or other robust methods. Results Structure counts by independent analysts were reasonably consistent. Between one and 11 occupancy reports were available per site and most of these reported people per household rather than per structure. The imagery-based method had a precision relative to reference population figures of Conclusions In settings with clearly distinguishable individual structures, the remote, imagery-based method had reasonable accuracy for the purposes of rapid estimation, was simple and quick to implement, and would likely perform better in more current application. However, it may have insurmountable limitations in settings featuring connected buildings or shelters, a complex pattern of roofs and multi-level buildings. Based on these results, we discuss possible ways forward for the method’s development.

  2. A method for rapid measurement of intrarenal and other tissue pressures.

    Science.gov (United States)

    SWANN, H G; MONTGOMERY, A V; DAVIS, J C; MICKLE, E R

    1950-12-01

    A rapid method for measuring tissue pressures has been designed. A pressure of 250 mm. Hg is imposed on a manometer. Then the system is allowed to discharge into a needle cannula inserted in the tissue. The manometer forces out fluid (about 10 c.mm.) until the pressure within it is the same as that within the tissue. Records of the pressure changes are made. Each observation takes about a minute. The method gives results that are closely comparable with other reports of tissue pressures. With this method, the pressure in the following organs of dogs was found to be: kidney, 26 mm. Hg, cerebral cortex, 0 to 5 mm., muscle, 1 to 10 mm., spleen, S to 16 mm., subcutaneous tissue, 0 to 3 mm., and liver -2 to 14 mm. The reliability of the method was tested on the kidneys of decerebrate dogs. Measurements were found to be the same within narrow limits over a period of an hour; they were the same when taken simultaneously in different regions of the same kidney or in opposite kidneys. They were independent of the volume of fluid forced into the tissue. Similar pressures were observed with 1 or 5 or 10 holes bored in the shaft of the cannulating needle. The intrarenal pressure was also measured by inserting a needle cannula into the tissue and then allowing the pressure to reach equilibrium passively with a manometer. This method gave similar results. The intrarenal pressure has now found to be the same when measured by three different technics.

  3. Improved Savitzky-Golay-method-based fluorescence subtraction algorithm for rapid recovery of Raman spectra.

    Science.gov (United States)

    Chen, Kun; Zhang, Hongyuan; Wei, Haoyun; Li, Yan

    2014-08-20

    In this paper, we propose an improved subtraction algorithm for rapid recovery of Raman spectra that can substantially reduce the computation time. This algorithm is based on an improved Savitzky-Golay (SG) iterative smoothing method, which involves two key novel approaches: (a) the use of the Gauss-Seidel method and (b) the introduction of a relaxation factor into the iterative procedure. By applying a novel successive relaxation (SG-SR) iterative method to the relaxation factor, additional improvement in the convergence speed over the standard Savitzky-Golay procedure is realized. The proposed improved algorithm (the RIA-SG-SR algorithm), which uses SG-SR-based iteration instead of Savitzky-Golay iteration, has been optimized and validated with a mathematically simulated Raman spectrum, as well as experimentally measured Raman spectra from non-biological and biological samples. The method results in a significant reduction in computing cost while yielding consistent rejection of fluorescence and noise for spectra with low signal-to-fluorescence ratios and varied baselines. In the simulation, RIA-SG-SR achieved 1 order of magnitude improvement in iteration number and 2 orders of magnitude improvement in computation time compared with the range-independent background-subtraction algorithm (RIA). Furthermore the computation time of the experimentally measured raw Raman spectrum processing from skin tissue decreased from 6.72 to 0.094 s. In general, the processing of the SG-SR method can be conducted within dozens of milliseconds, which can provide a real-time procedure in practical situations.

  4. A Rapid Method for Quantifying Viable Mycobacterium avium subsp. paratuberculosis in Cellular Infection Assays

    Science.gov (United States)

    Pooley, Hannah B.; de Silva, Kumudika; Purdie, Auriol C.; Begg, Douglas J.; Whittington, Richard J.

    2016-01-01

    ABSTRACT Determining the viability of bacteria is a key outcome of in vitro cellular infection assays. Currently, this is done by culture, which is problematic for fastidious slow-growing bacteria such as Mycobacterium avium subsp. paratuberculosis, where it can take up to 4 months to confirm growth. This study aimed to identify an assay that can rapidly quantify the number of viable M. avium subsp. paratuberculosis cells in a cellular sample. Three commercially available bacterial viability assays along with a modified liquid culture method coupled with high-throughput quantitative PCR growth detection were assessed. Criteria for assessment included the ability of each assay to differentiate live and dead M. avium subsp. paratuberculosis organisms and their accuracy at low bacterial concentrations. Using the culture-based method, M. avium subsp. paratuberculosis growth was reliably detected and quantified within 2 weeks. There was a strong linear association between the 2-week growth rate and the initial inoculum concentration. The number of viable M. avium subsp. paratuberculosis cells in an unknown sample was quantified based on the growth rate, by using growth standards. In contrast, none of the commercially available viability assays were suitable for use with samples from in vitro cellular infection assays. IMPORTANCE Rapid quantification of the viability of Mycobacterium avium subsp. paratuberculosis in samples from in vitro cellular infection assays is important, as it allows these assays to be carried out on a large scale. In vitro cellular infection assays can function as a preliminary screening tool, for vaccine development or antimicrobial screening, and also to extend findings derived from experimental animal trials. Currently, by using culture, it takes up to 4 months to obtain quantifiable results regarding M. avium subsp. paratuberculosis viability after an in vitro infection assay; however, with the quantitative PCR and liquid culture method

  5. A Simple and Rapid Identification Method for Mycobacterium bovis BCG with Loop-Mediated Isothermal Amplification.

    Directory of Open Access Journals (Sweden)

    Yuji Kouzaki

    Full Text Available Bacillus Calmette-Guérin (BCG is widely used as a live attenuated vaccine against Mycobacterium tuberculosis and is an agent for standard prophylaxis against the recurrence of bladder cancer. Unfortunately, it can cause severe infectious diseases, especially in immunocompromised patients, and the ability to immediately distinguish BCG from other M. tuberculosis complexes is therefore important. In this study, we developed a simple and easy-to-perform identification procedure using loop-mediated amplification (LAMP to detect deletions within the region of difference, which is deleted specifically in all M. bovis BCG strains. Reactions were performed at 64 °C for 30 min and successful targeted gene amplifications were detected by real-time turbidity using a turbidimeter and visual inspection of color change. The assay had an equivalent detection limit of 1.0 pg of genomic DNA using a turbidimeter whereas it was 10 pg with visual inspection, and it showed specificity against 49 strains of 44 pathogens, including M. tuberculosis complex. The expected LAMP products were confirmed through identical melting curves in real-time LAMP procedures. We employed the Procedure for Ultra Rapid Extraction (PURE kit to isolate mycobacterial DNA and found that the highest sensitivity limit with a minimum total cell count of mycobacterium (including DNA purification with PURE was up to 1 × 10(3 cells/reaction, based on color changes under natural light with FDA reagents. The detection limit of this procedure when applied to artificial serum, urine, cerebrospinal fluid, and bronchoalveolar lavage fluid samples was also about 1 × 10(3 cells/reaction. Therefore, this substitute method using conventional culture or clinical specimens followed by LAMP combined with PURE could be a powerful tool to enable the rapid identification of M. bovis BCG as point-of-care testing. It is suitable for practical use not only in resource-limited situations, but also in any

  6. Development of a fluorescence in situ hybridization (FISH) method for rapid detection of Ulva prolifera

    Science.gov (United States)

    Zhang, Qing-Chun; Liu, Qing; Kang, Zhen-Jun; Yu, Ren-Cheng; Yan, Tian; Zhou, Ming-Jiang

    2015-09-01

    Large-scale green tides have occurred consecutively since 2007 in the Yellow Sea (YS), China. The dominant causative species of the green tides has been identified as Ulva prolifera. The origin of green tides in the YS has been traced back to the Subei Shoal based on the results of remote-sensing, numerical simulations and field investigations. However, it is difficult to study the early development of green tides in the Subei Shoal because of the mixture of multiple green algae and the morphological diversity of U. prolifera when under variable environmental conditions. In this study, a rapid and accurate fluorescence in situ hybridization (FISH) method was developed to detect U. prolifera from the community of green algae targeting the 5S rDNA spacer region of U. prolifera. Two specific probes, 5S-1 and 5S-2, were designed based on the sequences of the 5S rDNA spacer regions of U. prolifera, Ulva linza and Ulva flexuosa. Specificity of the FISH method was tested using the six species of green algae commonly occurring in the Subei Shoal, including U. prolifera, U. linza, U. flexuosa, Ulva compressa, Ulva pertusa and Blidingia sp. The results showed that only U. prolifera could be labeled with both probes. Probe 5S-1, which showed a much higher labeling efficiency on U. prolifera, was ultimately selected as the probe for the FISH detection. The sample preparation method was optimized, particularly for the mature green algae, by the addition of cellulase and proteinase K in the pre-hybridization solution. Labeling efficiency with the probe 5S-1 reached 96% on average under the optimized conditions. The successful development of the FISH method has been applied to qualitative and quantitative analysis of field samples collected from the YS, and the results indicate a potential use in future green algae studies.

  7. A rapid method for the sequential separation of polonium, plutonium, americium and uranium in drinking water.

    Science.gov (United States)

    Lemons, B; Khaing, H; Ward, A; Thakur, P

    2018-02-06

    A new sequential separation method for the determination of polonium and actinides (Pu, Am and U) in drinking water samples has been developed that can be used for emergency response or routine water analyses. For the first time, the application of TEVA chromatography column in the sequential separation of polonium and plutonium has been studied. This method utilizes a rapid Fe +3 co-precipitation step to remove matrix interferences, followed by plutonium oxidation state adjustment to Pu 4+ and an incubation period of ~ 1 h at 50-60 °C to allow Po 2+ to oxidize to Po 4+ . The polonium and plutonium were then separated on a TEVA column, while separation of americium from uranium was performed on a TRU column. After separation, polonium was micro-precipitated with copper sulfide (CuS), while actinides were micro co-precipitated using neodymium fluoride (NdF 3 ) for counting by the alpha spectrometry. The method is simple, robust and can be performed quickly with excellent removal of interferences, high chemical recovery and very good alpha peak resolution. The efficiency and reliability of the procedures were tested by using spiked samples. The effect of several transition metals (Cu 2+ , Pb 2+ , Fe 3+ , Fe 2+ , and Ni 2+ ) on the performance of this method were also assessed to evaluate the potential matrix effects. Studies indicate that presence of up to 25 mg of these cations in the samples had no adverse effect on the recovery or the resolution of polonium alpha peaks. Copyright © 2018 Elsevier Ltd. All rights reserved.

  8. Veterinary drug residues in meat: Concerns and rapid methods for detection.

    Science.gov (United States)

    Reig, Milagro; Toldrá, Fidel

    2008-01-01

    The use of substances having hormonal or thyreostatic action as well as β-agonists is banned in the European Union. However, sometimes forbidden drugs may be added to feeds for illegal administration to farm animals for promoting increased muscle development or increased water retention and thus obtain an economical benefit. The result is a fraudulent overweight of meat but, what is worse, residues of these substances may remain in meat and may pose a real threat to the consumer either through exposure to the residues, transfer of antibiotic resistance or allergy risk. This has exerted a great concern among European consumers. The control of the absence of these forbidden substances in animal foods and feeds is regulated in the European Union by Directive96/23/EC on measures to monitor certain substances and residues in live animals and animal products. Analytical methodology, including criteria for identification and confirmation, for the monitoring of compliance was also given in Decisions 93/256/EEC and 93/257/EEC. More recently, Decision 2002/657/EC provided rules for the analytical methods to be used in testing of official samples. A crucial step is the screening of veterinary drug residues in live animals, feeds and animal products in view of the remarkable number of samples and large variety of residues to be analysed. In recent years, different rapid methods having easy performance, high sensitivity and high throughput have been proposed and are being extensively used. These methods as well as other new methods are reviewed in this manuscript.

  9. TranScreen-N: Method for rapid screening of trans-ungual drug delivery enhancers.

    Science.gov (United States)

    Murthy, S Narasimha; Vaka, Siva Ram Kiran; Sammeta, Srinivasa Murthy; Nair, Anroop B

    2009-11-01

    Topical monotherapy of nail diseases such as onychomycosis and nail psoriasis has been less successful due to poor permeability of the human nail plate to topically administered drugs. Chemical enhancers are utilized to improve the drug delivery across the nail plate. Choosing the most effective chemical enhancers for the given drug and formulation is highly critical in determining the efficacy of topical therapy of nail diseases. Screening the large pool of enhancers using currently followed diffusion cell experiments would be tedious and expensive. The main objective of this study is to develop TranScreen-N, a high throughput method of screening trans-ungual drug permeation enhancers. It is a rapid microwell plate based method which involves two different treatment procedures; the simultaneous exposure treatment and the sequential exposure treatment. In the present study, several chemicals were evaluated by TranScreen-N and by diffusion studies in the Franz diffusion cell (FDC). Good agreement of in vitro drug delivery data with TranScreen-N data provided validity to the screening technique. In TranScreen-N technique, the enhancers can be grouped according to whether they need to be applied before or simultaneously with drugs (or by either procedures) to enhance the drug delivery across the nail plate. TranScreen-N technique can significantly reduce the cost and duration required to screen trans-ungual drug delivery enhancers. (c) 2009 Wiley-Liss, Inc. and the American Pharmacists Association

  10. A rapid method to authenticate vegetable oils through surface-enhanced Raman scattering

    Science.gov (United States)

    Lv, Ming Yang; Zhang, Xin; Ren, Hai Rui; Liu, Luo; Zhao, Yong Mei; Wang, Zheng; Wu, Zheng Long; Liu, Li Min; Xu, Hai Jun

    2016-03-01

    Vegetable oils are essential in our daily diet. Among various vegetable oils, the major difference lies in the composition of fatty acids, including unsaturated fatty acids (USFA) and saturated fatty acids (SFA). USFA include oleic acid (OA), linoleic acid (LA), and α-linolenic acid (ALA), while SFA are mainly palmitic acid (PA). In this study, the most typical and abundant USFA present with PA in vegetable oils were quantified. More importantly, certain proportional relationships between the integrated intensities of peaks centered at 1656 cm-1 (S1656) in the surface-enhanced Raman scattering spectra of different USFA were confirmed. Therefore, the LA or ALA content could be converted into an equivalent virtual OA content enabling the characterization of the USFA content in vegetable oils using the equivalent total OA content. In combination with the S1656 of pure OA and using peanut, sesame, and soybean oils as examples, the ranges of S1656 corresponding to the National Standards of China were established to allow the rapid authentication of vegetable oils. Gas chromatograph-mass spectrometer analyses verified the accuracy of the method, with relative errors of less than 5%. Moreover, this method can be extended to other detection fields, such as diseases.

  11. New multiplex PCR methods for rapid screening of genetically modified organisms in foods.

    Science.gov (United States)

    Datukishvili, Nelly; Kutateladze, Tamara; Gabriadze, Inga; Bitskinashvili, Kakha; Vishnepolsky, Boris

    2015-01-01

    We present novel multiplex PCR methods for rapid and reliable screening of genetically modified organisms (GMOs). New designed PCR primers targeting four frequently used GMO specific sequences permitted identification of new DNA markers, in particular 141 bp fragment of cauliflower mosaic virus (CaMV) 35S promoter, 224 bp fragment of Agrobacterium tumefaciens nopaline synthase (NOS) terminator, 256 bp fragment of 5-enolppyruvylshikimate-phosphate synthase (epsps) gene and 258 bp fragment of Cry1Ab delta-endotoxin (cry1Ab) gene for GMO screening. The certified reference materials containing Roundup Ready soybean (RRS) and maize MON 810 were applied for the development and optimization of uniplex and multiplex PCR systems. Evaluation of amplification products by agarose gel electrophoresis using negative and positive controls confirmed high specificity and sensitivity at 0.1% GMO for both RRS and MON 810. The fourplex PCR was developed and optimized that allows simultaneous detection of three common transgenic elements, such as: CaMV 35S promoter, NOS terminator, epsps gene together with soybean-specific lectin gene. The triplex PCR developed enables simultaneous identification of transgenic elements, such as: 35S promoter and cry1Ab gene together with maize zein gene. The analysis of different processed foods demonstrated that multiplex PCR methods developed in this study are useful for accurate and fast screening of GM food products.

  12. A simple and rapid method for optical visualization and quantification of bacteria on textiles

    Science.gov (United States)

    Stiefel, Philipp; Schneider, Jana; Amberg, Caroline; Maniura-Weber, Katharina; Ren, Qun

    2016-01-01

    To prevent bacterial contamination on textiles and the associated undesired effects different biocidal coatings have been investigated and applied. However, due to health and environmental concerns anti-adhesive coatings preventing the binding of bacteria would be favored. To develop such anti-adhesive coatings simple assays for reliable and fast screening are beneficial. Here an easy-to-handle, robust and rapid assay to assess bacteria on textiles utilizing a tetrazolium salt was reported. The assay allowed direct eye visualization of the color change of the textiles containing bacteria, facilitating fast screening. Quantification of the adhered bacteria could be done by generating standard curves which correlate the staining intensity to cell numbers. An additional advantage of the described assay is that with the same detection method anti-adhesive and biocidal effects can be investigated. The method was applied to different coatings, using Pseudomonas aeruginosa and Staphylococcus aureus as model organisms. The detection limit was found to be between 2.5 * 106 and 9.4 * 108 for P. aeruginosa and between 1 * 106 and 3.3 * 108 for S. aureus. The anti-adhesive coating PLUMA was demonstrated to reduce bacterial adhesion without killing them, whereas the biocidal coating TH22-27 caused a clear reduction in the number of viable cells. PMID:28004762

  13. The rapid interphase chromosome assay (RICA implementation: comparison with other PCC methods

    Directory of Open Access Journals (Sweden)

    Sommer Sylwester

    2015-12-01

    Full Text Available A report is presented on the advantages of the rapid interphase chromosome assay (RICA and the difficulties that may be met while implementing this method for application in biological dosimetry. The RICA test can be applied on unstimulated human lymphocytes; this is an advantage in comparison with the dicentric chromosomes or micronucleus tests. In the former two tests, stimulated lymphocytes are examined and hence, 48 h more are needed to obtain cells traversing the cell cycle. Due to the use of unstimulated nondividing cells, higher numbers of cells are available for RICA analysis than for dicentric chromosomes or micronuclei tests. Moreover, the method can be applied after exposure to ionizing radiation doses in excess of 5 Gy. Such doses cause a significant cell cycle delay or result in the loss of G2 phase and mitotic cells because of apoptosis. Therefore, the traditional biodosimetry based on the evaluation of the incidence of damage to chromosomes is very difficult to carry out. This is due to the lack of an adequate number of mitotic cells for analysis. RICA is free of this disadvantage. An automatic microscope can be used to retrieve cell images; automatic image analysis can also be used.

  14. Development of a Rapid and Simple Method to Remove Polyphenols from Plant Extracts

    Directory of Open Access Journals (Sweden)

    Imali Ranatunge

    2017-01-01

    Full Text Available Polyphenols are secondary metabolites of plants, which are responsible for prevention of many diseases. Polyvinylpolypyrrolidone (PVPP has a high affinity towards polyphenols. This method involves the use of PVPP column to remove polyphenols under centrifugal force. Standards of gallic acid, epigallocatechin gallate, vanillin, and tea extracts (Camellia sinensis were used in this study. PVPP powder was packed in a syringe with different quantities. The test samples were layered over the PVPP column and subjected to centrifugation. Supernatant was tested for the total phenol content. The presence of phenolic compounds and caffeine was screened by HPLC and measuring the absorbance at 280. The antioxidant capacity of standards and tea extracts was compared with the polyphenol removed fractions using DPPH scavenging assay. No polyphenols were found in polyphenolic standards or tea extracts after PVPP treatment. The method described in the present study to remove polyphenols is simple, inexpensive, rapid, and efficient and can be employed to investigate the contribution of polyphenols present in natural products to their biological activity.

  15. A Rapid and Low-Cost Nonlithographic Method to Fabricate Biomedical Microdevices for Blood Flow Analysis

    Directory of Open Access Journals (Sweden)

    Elmano Pinto

    2014-12-01

    Full Text Available Microfluidic devices are electrical/mechanical systems that offer the ability to work with minimal sample volumes, short reactions times, and have the possibility to perform massive parallel operations. An important application of microfluidics is blood rheology in microdevices, which has played a key role in recent developments of lab-on-chip devices for blood sampling and analysis. The most popular and traditional method to fabricate these types of devices is the polydimethylsiloxane (PDMS soft lithography technique, which requires molds, usually produced by photolithography. Although the research results are extremely encouraging, the high costs and time involved in the production of molds by photolithography is currently slowing down the development cycle of these types of devices. Here we present a simple, rapid, and low-cost nonlithographic technique to create microfluidic systems for biomedical applications. The results demonstrate the ability of the proposed method to perform cell free layer (CFL measurements and the formation of microbubbles in continuous blood flow.

  16. A rapid and sensitive spectrophotometric method for the determination of benzoyl peroxide in wheat flour samples

    Directory of Open Access Journals (Sweden)

    Kraingkrai Ponhong

    2015-12-01

    Full Text Available A simple, rapid, and sensitive spectrophotometric method for the determination of benzoyl peroxide (BPO in wheat flour samples was developed. The detection principle is based on BPO reacted with 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS to obtain a blue-green colored product that was detected at 415 nm by spectrophotometry. The effect of factors influencing the color reaction was investigated. Under the selected conditions, the linear range for quantification of BPO was observed between 0.2–1.0 mg L−1 with r2 = 0.998. The limit of detection (LOD was 0.025 mg L−1. The developed method obtained superior precision (relative standard deviation < 2% using 11 repeatability at 0.2 mg L−1, 0.6 mg L−1, and 0.8 mg L−1. The proposed methodology was successfully applied to determine BPO in wheat flour samples.

  17. New multiplex PCR methods for rapid screening of genetically modified organisms in foods

    Directory of Open Access Journals (Sweden)

    Nelly eDatukishvili

    2015-07-01

    Full Text Available We present novel multiplex PCR methods for rapid and reliable screening of genetically modified organisms (GMOs. New designed PCR primers targeting four frequently used GMO specific sequences permitted identification of new DNA markers, in particular 141 bp fragment of cauliflower mosaic virus (CaMV 35S promoter, 224 bp fragment of Agrobacterium tumefaciens nopaline synthase (NOS terminator, 256 bp fragment of 5-enolppyruvylshikimate-phosphate synthase (epsps gene and 258 bp fragment of Cry1Ab delta-endotoxin (cry1Ab gene for GMO screening. The certified reference materials containing Roundup Ready soybean (RRS and maize MON 810 were applied for the development and optimization of uniplex and multiplex PCR systems. Evaluation of amplification products by agarose gel electrophoresis using negative and positive controls confirmed high specificity and sensitivity at 0.1% GMO for both RRS and MON 810. The fourplex PCR was developed and optimized that allows simultaneous detection of three common transgenic elements, such as: CaMV 35S promoter, NOS terminator, epsps gene together with soybean-specific lectin gene. The triplex PCR developed enables simultaneous identification of transgenic elements, such as: 35S promoter and cry1Ab gene together with maize zein gene. The analysis of different processed foods demonstrated that multiplex PCR methods developed in this study are useful for accurate and fast screening of GM food products.

  18. Regional distribution of methionine adenosyltransferase in rat brain as measured by a rapid radiochemical method

    Energy Technology Data Exchange (ETDEWEB)

    Hiemke, C.; Ghraf, R.

    1981-09-01

    The distribution of methionine adenosyltransferase (MAT) in the CNS of the rat was studied by use of a rapid, sensitive and specific radiochemical method. The S-adenosyl-(methyl-/sup 14/C)L-methionine ((/sup 14/C)SAM) generated by adenosyl transfer from ATP to (methyl-/sup 14/C)L-methionine is quantitated by use of a SAM-consuming transmethylation reaction. Catechol O-methyltransferase (COMT), prepared from rat liver, transfers the methyl-/sup 14/C group of SAM to 3,4-dihydroxybenzoic acid. The /sup 14/C-labelled methylation products, vanillic acid and isovanillic acid, are separated from unreacted methionine by solvent extraction and quantitated by liquid scintillation counting. Compared to other methods of MAT determination, which include separation of generated SAM from methionine by ion-exchange chromatography, the assay described exhibited the same high degree of specificity and sensitivity but proved to be less time consuming. MAT activity was found to be uniformly distributed between various brain regions and the pituitary gland of adult male rats. In the pineal gland the enzyme activity is about tenfold higher.

  19. Method and apparatus using selected superparamagnetic labels for rapid quantification of immunochromatographic tests

    Directory of Open Access Journals (Sweden)

    Mika PA Laitinen

    2009-04-01

    Full Text Available Mika PA Laitinen1, Jari Salmela2, Leona Gilbert1, Risto Kaivola1, Topi Tikkala2, Christian Oker-Blom1, Jukka Pekola3, Matti Vuento11Department of Biological and Environmental Science; 2Department of Physics, University of Jyväskylä, Jyväskylä, Finland; 3Low Temperature Laboratory, Helsinki University of Technology, Helsinki, FinlandAbstract: A rapid method and instrumentation for quantification of immunochromatographic tests (ICT are described. The principle and performance of the method was demonstrated by measuring the levels of human chorionic gonadotropin (hCG present in urine. The test format was a sandwich assay using two distinct monoclonal antibodies directed against hCG. The first anti-hCG antibody was labeled with superparamagnetic particles whereas the second was immobilized as a narrow detection zone on a porous membrane. The human urine sample was mixed with superparamagnetic particles coated with the first anti-hCG antibody, and the mixture was allowed to migrate past the detection zone containing the second anti-hCG antibody. Capillary forces facilitated migration of the immune complexes along the porous membrane. The amount of superparamagnetic particle-labelled monoclonal anti-hCG bound to the detection zone was directly proportional to the amount of hCG present in the sample as detected by measuring magnetization in the detector coil. The method had a practical detection limit of 20 U/l (54 nM of hCG per 5 μl of human urine and a linear range of three decades from 20 U/l to 10 000 U/l. In addition, the analysis was completed within less than 10 minutes. Thus, the test format should be suitable for fast detection and monitoring of a large variety of clinically important parameters and analytes.Keywords: affinity, biosensor, hCG, immunochromatography, magnetization, superparamagnetic

  20. Rapid methods to assess sanitizing efficacy of benzalkonium chloride to Listeria monocytogenes biofilms.

    Science.gov (United States)

    Romanova, Nadya A; Gawande, Purushottam V; Brovko, Lubov Y; Griffiths, Mansel W

    2007-12-01

    Different methods were used to investigate biofilm growth including crystal violet staining, ATP bioluminescence and total viable count. Seven strains of Listeria monocytogenes and 8 of their derivative strains were screened for their capacity to form biofilms. Both adaptation to benzalkonium chloride (BC) and curing of plasmids did not significantly affect biofilm-forming ability. The strains of L. monocytogenes belonging to serotype 1 formed biofilms significantly better as compared to serotype 4 (P=0.0003). To estimate the efficacy of BC for biofilm elimination the best and the poorest biofilm-formers were used (C719 and LJH 381). It was observed that, L. monocytogenes strain C719 in biofilms is at least 1000 times more resistant to BC than in planktonic form. Cells present in biofilms were shown to recover and grow after BC treatment thus providing a source of recontamination. It was shown that ATP bioluminescence provides good correlation with bacterial counts of L. monocytogenes in biofilms. Staining with crystal violet, on the contrary, did not correlate with bacterial growth in biofilms in the presence of high concentrations of BC but provided information on the concentration of bacterial cells, both live and dead, attached to the surface. ATP bioluminescence was found to be a reliable method for rapid estimation of the efficacy of sanitizers for biofilm disinfection. Crystal violet staining, on the other hand, was shown to be a suitable method to monitor removal of biofilms. Our investigation showed that for Listeria biofilms concentrations of BC higher then 10 mg/ml should be applied for at least 30 min to kill almost all the live cells in biofilms. However, this concentration was still not enough to remove biofilms from the surface of plastic.

  1. Development of a rapid, one-step screening method for the isolation of presumptive proteolytic enterococci.

    Science.gov (United States)

    Graham, Ken; Rea, Rosemary; Simpson, Paul; Stack, Helena

    2017-01-01

    Enterococci show higher proteolytic activities than other lactic acid bacteria and thus have received considerable attention in scientific literature in recent years. Proteolytic enzymes of enterococci have warranted the use of some species as starter, adjuncts or protective cultures and as probiotics, while in some strains they have also been linked with virulence. Consequently, the isolation and identification of proteolytic enterococci is becoming of increasing interest and importance. However, current screening methods for proteolytic enterococci can be time consuming, requiring a two-step procedure which may take up to 96h. This study describes a method, utilising Kanamycin Skim Milk Aesculin Azide (KSMEA) agar, for the isolation of proteolytic enterococci in one-step, thereby significantly reducing screening time. KSMEA combines the selective properties of Kanamycin Aesculin Azide Agar (KAA) with skim milk powder for the detection of proteolytic enterococci. Enterococci produced colonies with a black halo on KSMEA which were accompanied by a zone of clearing in the media when enterococci were proteolytic. KSMEA medium retained the selectivity of KAA, while proteolytic enterococci were easily distinguished from non-proteolytic enterococci when two known strains were propagated on KSMEA. KSMEA also proved effective at isolating and detecting enterococci in raw milk, faeces and soil. Isolates recovered from the screen were confirmed as enterococci using genus-specific primers. Proteolytic enterococci were present in the raw milk sample only and were easily distinguishable from non-proteolytic enterococci and other microorganisms. Therefore, KSMEA provides a rapid, one-step screening method for the isolation of presumptive proteolytic enterococci. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Improvements in fast-neutron spectroscopy methods (1961); Amelioration des methodes de spectrometrie des neutrons rapides (1961)

    Energy Technology Data Exchange (ETDEWEB)

    Cambou, F. [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires

    1961-02-15

    This research aimed at improving fast-neutron electronic detectors based on n-p elastic scattering. The first part concerns proportional counters; careful constructional methods have made it possible to plot mono-energetic neutron spectra in the range 700 keV - 3 MeV with a resolution of 7 per cent. The second part concerns scintillation counters: an organic scintillator and an inorganic scintillator covered with a thin layer of a scattering agent. An exact study of the types of scintillation has made it possible to develop efficient discriminator circuits. Different neutron spectra plotted in the presence of a strong gamma background are presented. The last part deals with the development of form discrimination methods for the study, in the actual beam, of the elastic scattering of 14.58 MeV electrons. With hydrogen, the distribution f ({phi}) of the recoil protons is f({phi}) = 1 + 0.034 cos {phi} + 0.042 cos{sup 2} {phi}. With tritium the scattering is strongly anisotropic; the curve representing the variation of the differential cross-section for the elastic scattering in the centre of mass system is obtained with a target containing 1 cm{sup 3} of tritium. (author) [French] Le travail a porte sur l'amelioration des detecteurs electroniques de neutrons rapides bases sur la diffusion elastique n-p. La premiere partie est relative aux compteurs proportionnels; des methodes soignees de fabrication ont permis des traces de spectres de neutrons monoenergetiques dans le domaine 700 keV - 3 MeV avec une resolution de 7 pour cent. La deuxieme partie est relative au compteur a scintillations; scintillateur organique et scintillateur mineral recouvert d'un diffuseur mince. Une etude precise des formes de scintillations a permis la mise au point de circuits discriminateurs efficaces. Differents spectres de neutrons traces en presence d'un fond gamma intense sont presentes. La derniere partie est relative a la mise en oeuvre des methodes de discrimination de

  3. Development of a rapid method for the quantitative determination of deoxynivalenol using Quenchbody

    Energy Technology Data Exchange (ETDEWEB)

    Yoshinari, Tomoya [Division of Microbiology, National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501 (Japan); Ohashi, Hiroyuki; Abe, Ryoji; Kaigome, Rena [Biomedical Division, Ushio Inc., 1-12 Minamiwatarida-cho, Kawasaki-ku, Kawasaki 210-0855 (Japan); Ohkawa, Hideo [Research Center for Environmental Genomics, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501 (Japan); Sugita-Konishi, Yoshiko, E-mail: y-konishi@azabu-u.ac.jp [Department of Food and Life Science, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa 252-5201 (Japan)

    2015-08-12

    Quenchbody (Q-body) is a novel fluorescent biosensor based on the antigen-dependent removal of a quenching effect on a fluorophore attached to antibody domains. In order to develop a method using Q-body for the quantitative determination of deoxynivalenol (DON), a trichothecene mycotoxin produced by some Fusarium species, anti-DON Q-body was synthesized from the sequence information of a monoclonal antibody specific to DON. When the purified anti-DON Q-body was mixed with DON, a dose-dependent increase in the fluorescence intensity was observed and the detection range was between 0.0003 and 3 mg L{sup −1}. The coefficients of variation were 7.9% at 0.003 mg L{sup −1}, 5.0% at 0.03 mg L{sup −1} and 13.7% at 0.3 mg L{sup −1}, respectively. The limit of detection was 0.006 mg L{sup −1} for DON in wheat. The Q-body showed an antigen-dependent fluorescence enhancement even in the presence of wheat extracts. To validate the analytical method using Q-body, a spike-and-recovery experiment was performed using four spiked wheat samples. The recoveries were in the range of 94.9–100.2%. The concentrations of DON in twenty-one naturally contaminated wheat samples were quantitated by the Q-body method, LC-MS/MS and an immunochromatographic assay kit. The LC-MS/MS analysis showed that the levels of DON contamination in the samples were between 0.001 and 2.68 mg kg{sup −1}. The concentrations of DON quantitated by LC-MS/MS were more strongly correlated with those using the Q-body method (R{sup 2} = 0.9760) than the immunochromatographic assay kit (R{sup 2} = 0.8824). These data indicate that the Q-body system for the determination of DON in wheat samples was successfully developed and Q-body is expected to have a range of applications in the field of food safety. - Highlights: • A rapid method for quantitation of DON using Q-body has been developed. • A recovery test using the anti-DON Q-body was performed. • The concentrations of DON in wheat

  4. A rapid method for detection of five known mutations associated with aminoglycoside-induced deafness

    Directory of Open Access Journals (Sweden)

    Greinwald John H

    2009-01-01

    Full Text Available Abstract Background South Africa has one of the highest incidences of multidrug-resistant tuberculosis (MDR-TB in the world. Concomitantly, aminoglycosides are commonly used in this country as a treatment against MDR-TB. To date, at least five mutations are known to confer susceptibility to aminoglycoside-induced hearing loss. The aim of the present study was to develop a rapid screening method to determine whether these mutations are present in the South African population. Methods A multiplex method using the SNaPshot technique was used to screen for five mutations in the MT-RNR1 gene: A1555G, C1494T, T1095C, 961delT+C(n and A827G. A total of 204 South African control samples, comprising 98 Mixed ancestry and 106 Black individuals were screened for the presence of the five mutations. Results A robust, cost-effective method was developed that detected the presence of all five sequence variants simultaneously. In this pilot study, the A1555G mutation was identified at a frequency of 0.9% in the Black control samples. The 961delT+C(n variant was present in 6.6% of the Black controls and 2% of the Mixed ancestry controls. The T1095C, C1494T and A827G variants were not identified in any of the study participants. Conclusion The frequency of 0.9% for the A1555G mutation in the Black population in South Africa is of concern given the high incidence of MDR-TB in this particular ethnic group. Future larger studies are warranted to determine the true frequencies of the aminoglycoside deafness mutations in the general South African population. The high frequencies of the 961delT+C(n variant observed in the controls suggest that this change is a common non-pathogenic polymorphism. This genetic method facilitates the identification of individuals at high risk of developing hearing loss prior to the start of aminoglycoside therapy. This is important in a low-resource country like South Africa where, despite their adverse side-effects, aminoglycosides will

  5. A Rapid and Simple TLC-Densitometric Method for Assay of Clobetasol Propionate in Topical Solution.

    Science.gov (United States)

    Dolowy, Malgorzata; Kozik, Violetta; Bak, Andrzej; Jampilek, Josef; Barbusinski, Krzysztof; Thomas, Maciej; Pyka-Pajak, Alina

    2017-11-03

    A rapid, simple to use and low-cost thin-layer chromatographic procedure in normal phase system with densitometric detection at 246 nm was carefully validated according to the International Conference on Harmonisation (ICH) guidelines for assay of clobetasol propionate in topical solution containing clobetasol propionate in quantity 0.50 mg/mL. The adopted thin-layer chromatographic (TLC)-densitometric procedure could effectively separate clobetasol propionate from its related compound, namely clobetasol. It is linear for clobetasol propionate in the range of 0.188 ÷ 5 µg/spot. The limit of detection (LOD) and limit of quantification (LOQ) value is 0.061 and 0.186 µg/spot, respectively. Accuracy of proposed procedure was evaluated by recovery test. The mean recovery of studied clobetasol propionate ranges from 98.7 to 101.0%. The coefficient of variation (CV, %) obtained during intra-day and inter-day studies, which was less than 2% (0.40 ÷ 1.17%), confirms the precision of described method. The assay value of clobetasol propionate is consistent with the pharmacopoeial requirements. In conclusion, it can be suitable as a simple and economic procedure for routine quality control laboratories of clobetasol propionate in topical solution.

  6. Hydrodynamic Voltammetry as a Rapid and Simple Method for Evaluating Soil Enzyme Activities

    Directory of Open Access Journals (Sweden)

    Kazuto Sazawa

    2015-03-01

    Full Text Available Soil enzymes play essential roles in catalyzing reactions necessary for nutrient cycling in the biosphere. They are also sensitive indicators of ecosystem stress, therefore their evaluation is very important in assessing soil health and quality. The standard soil enzyme assay method based on spectroscopic detection is a complicated operation that requires the removal of soil particles. The purpose of this study was to develop a new soil enzyme assay based on hydrodynamic electrochemical detection using a rotating disk electrode in a microliter droplet. The activities of enzymes were determined by measuring the electrochemical oxidation of p-aminophenol (PAP, following the enzymatic conversion of substrate-conjugated PAP. The calibration curves of β-galactosidase (β-gal, β-glucosidase (β-glu and acid phosphatase (AcP showed good linear correlation after being spiked in soils using chronoamperometry. We also performed electrochemical detection using real soils. Hydrodynamic chronoamperometry can be used to assess the AcP in soils, with a detection time of only 90 s. Linear sweep voltammetry was used to measure the amount of PAP released from β-gal and β-glu by enzymatic reaction after 60 min. For the assessment of soil enzymes, the results of hydrodynamic voltammetry assay compared favorably to those using a standard assay procedure, but this new procedure is more user-friendly, rapid and simple.

  7. An assessment of various blood collection and transfer methods used for malaria rapid diagnostic tests

    Directory of Open Access Journals (Sweden)

    Baik Fred

    2007-11-01

    Full Text Available Abstract Background Four blood collection and transfer devices commonly used for malaria rapid diagnostic tests (RDTs were assessed for their consistency, accuracy and ease of use in the hands of laboratory technicians and village health workers. Methods Laboratory technicians and village health workers collected blood from a finger prick using each device in random order, and deposited the blood either on filter paper or into a suitable casette-type RDT. Consistency and accuracy of volume delivered was determined by comparing the measurements of the resulting blood spots/heights with the measurements of laboratory-prepared pipetted standard volumes. The effect of varying blood volumes on RDT sensitivity and ease of use was also observed. Results There was high variability in blood volume collected by the devices, with the straw and the loop, the most preferred devices, usually transferring volumes greater than intended, while the glass capillary tube and the plastic pipette transferring less volume than intended or none at all. Varying the blood volume delivered to RDTs indicated that this variation is critical to RDT sensitivity only when the transferred volume is very low. Conclusion None of the blood transfer devices assessed performed consistently well. Adequate training on their use is clearly necessary, with more development efforts for improved designs to be used by remote health workers, in mind.

  8. Rapid characterization of transgenic and non-transgenic soybean oils by chemometric methods using NIR spectroscopy

    Science.gov (United States)

    Luna, Aderval S.; da Silva, Arnaldo P.; Pinho, Jéssica S. A.; Ferré, Joan; Boqué, Ricard

    Near infrared (NIR) spectroscopy and multivariate classification were applied to discriminate soybean oil samples into non-transgenic and transgenic. Principal Component Analysis (PCA) was applied to extract relevant features from the spectral data and to remove the anomalous samples. The best results were obtained when with Support Vectors Machine-Discriminant Analysis (SVM-DA) and Partial Least Squares-Discriminant Analysis (PLS-DA) after mean centering plus multiplicative scatter correction. For SVM-DA the percentage of successful classification was 100% for the training group and 100% and 90% in validation group for non transgenic and transgenic soybean oil samples respectively. For PLS-DA the percentage of successful classification was 95% and 100% in training group for non transgenic and transgenic soybean oil samples respectively and 100% and 80% in validation group for non transgenic and transgenic respectively. The results demonstrate that NIR spectroscopy can provide a rapid, nondestructive and reliable method to distinguish non-transgenic and transgenic soybean oils.

  9. New 16-plex PCR method for rapid detection of diarrheagenic Escherichia coli directly from stool samples.

    Science.gov (United States)

    Antikainen, J; Tarkka, E; Haukka, K; Siitonen, A; Vaara, M; Kirveskari, J

    2009-08-01

    A rapid 16-plex polymerase chain reaction (PCR) suitable for routine diagnostics of diarrheagenic Escherichia coli (EHEC, EIEC, EAEC, ETEC, and EPEC) was developed, validated with control strains, and tested with 250 diarrhoeal stool samples. The specificity was 100% when tested with 289 control bacterial strains, and the analytical sensitivity of automated DNA extraction directly from stool samples was made by boiling the bacterial culture (10(4)-10(5) colony forming units/ml). The assay design starting directly from extraction of stool DNA allowed same day analysis without compromising sensitivity and specificity, which makes it superior compared to PCR after culturing the bacteria. The 16-plex PCR method demonstrated high prevalence of diarrheagenic E. coli in stool samples of patients returning from abroad (39.0%) in contrast to the patients with no travel history (8.7%; p < 0.001). The high prevalence of diarrheagenic E. coli suggests that their screening should be part of normal diarrhoea diagnostics, at least in the leading diagnostic laboratories.

  10. A Simple and Rapid Method for Preparing a Cell-Free Bacterial Lysate for Protein Synthesis.

    Directory of Open Access Journals (Sweden)

    Nitzan Krinsky

    Full Text Available Cell-free protein synthesis (CFPS systems are important laboratory tools that are used for various synthetic biology applications. Here, we present a simple and inexpensive laboratory-scale method for preparing a CFPS system from E. coli. The procedure uses basic lab equipment, a minimal set of reagents, and requires less than one hour to process the bacterial cell mass into a functional S30-T7 extract. BL21(DE3 and MRE600 E. coli strains were used to prepare the S30-T7 extract. The CFPS system was used to produce a set of fluorescent and therapeutic proteins of different molecular weights (up to 66 kDa. This system was able to produce 40-150 μg-protein/ml, with variations depending on the plasmid type, expressed protein and E. coli strain. Interestingly, the BL21-based CFPS exhibited stability and increased activity at 40 and 45°C. To the best of our knowledge, this is the most rapid and affordable lab-scale protocol for preparing a cell-free protein synthesis system, with high thermal stability and efficacy in producing therapeutic proteins.

  11. HPLC method for rapidly following biodiesel fuel transesterification reaction progress using a core-shell column

    Energy Technology Data Exchange (ETDEWEB)

    Allen, Samuel J.; Ott, Lisa S. [California State University, Chico, CA (United States)

    2012-07-15

    There are a wide and growing variety of feedstocks for biodiesel fuel. Most commonly, these feedstocks contain triglycerides which are transesterified into the fatty acid alkyl esters (FAAEs) which comprise biodiesel fuel. While the tranesterification reaction itself is simple, monitoring the reaction progress and reaction products is not. Gas chromatography-mass spectrometry is useful for assessing the FAAE products, but does not directly address either the tri-, di-, or monoglycerides present from incomplete transesterification or the free fatty acids which may also be present. Analysis of the biodiesel reaction mixture is complicated by the solubility and physical property differences among the components of the tranesterification reaction mixture. In this contribution, we present a simple, rapid HPLC method which allows for monitoring all of the main components in a biodiesel fuel transesterification reaction, with specific emphasis on the ability to monitor the reaction as a function of time. The utilization of a relatively new, core-shell stationary phase for the HPLC column allows for efficient separation of peaks with short elution times, saving both time and solvent. (orig.)

  12. Lock-in thermography as a rapid and reproducible thermal characterization method for magnetic nanoparticles

    Science.gov (United States)

    Lemal, Philipp; Geers, Christoph; Monnier, Christophe A.; Crippa, Federica; Daum, Leopold; Urban, Dominic A.; Rothen-Rutishauser, Barbara; Bonmarin, Mathias; Petri-Fink, Alke; Moore, Thomas L.

    2017-04-01

    Lock-in thermography (LIT) is a sensitive imaging technique generally used in engineering and materials science (e.g. detecting defects in composite materials). However, it has recently been expanded for investigating the heating power of nanomaterials, such as superparamagnetic iron oxide nanoparticles (SPIONs). Here we implement LIT as a rapid and reproducible method that can evaluate the heating potential of various sizes of SPIONs under an alternating magnetic field (AMF), as well as the limits of detection for each particle size. SPIONs were synthesized via thermal decomposition and stabilized in water via a ligand transfer process. Thermographic measurements of SPIONs were made by stimulating particles of varying sizes and increasing concentrations under an AMF. Furthermore, a commercially available SPION sample was included as an external reference. While the size dependent heating efficiency of SPIONs has been previously described, our objective was to probe the sensitivity limits of LIT. For certain size regimes it was possible to detect signals at concentrations as low as 0.1 mg Fe/mL. Measuring at different concentrations enabled a linear regression analysis and extrapolation of the limit of detection for different size nanoparticles.

  13. A method for the rapid detection and identification of halo blight pathogen on common bean

    Directory of Open Access Journals (Sweden)

    Popović Tatjana

    2014-01-01

    Full Text Available A diagnostic method based on nested-PCR, followed by ELISA and conventional bacteriology tests, for the rapid and reliable detection of halo blight pathogen Pseudomonas savastanoi pv. phaseolicola (Psp collected from infected bean leaves and seeds is described. Psp formed white, small and flat colonies on nutrient agar medium, creamy white, flat and circular on Milk-Tween agar medium and light yellow, convex and shiny on modified sucrose peptone agar medium. Eighteen Gram-negative, catalase-positive and oxidase-negative strains were subjected to nested PCR with primers P 5.1/P 3.1 and P 5.2/P 3.2, which directed the amplification of the 450 bp target DNA fragment in all tested strains. According to the results of DAS- and PTA-ELISA with respect to reactivity to specific antibodies, all analyzed strains belonged to Psp bacterium. Pathogenicity was tested on bean pods and cotyledon leaves, on which greasy spots were formed. Psp did not cause hypersensitive reaction on the leaves of tobacco and geranium. Strains produced levan, fluorescent pigment, oxidative metabolism of glucose, did not reduce nitrate, did not produce indole and H2S, did not hydrolyze starch, gelatin and esculin; they produced acid from glucose, mannose, sucrose and glycerol, and did not produce acid from maltose, starch, esculin, dulcite, sorbitol, inositol and erythritol.

  14. Rapid validated HPTLC method for estimation of betulinic acid in Nelumbo nucifera (Nymphaeaceae) rhizome extract.

    Science.gov (United States)

    Mukherjee, Debajyoti; Kumar, N Satheesh; Khatua, Taraknath; Mukherjee, Pulok K

    2010-01-01

    Betulinic acid (pentacyclic triterpenoid) is an important marker component present in Nelumbo nucifera Gaertn. rhizome. N. nucifera rhizome has several medicinal uses including hypoglycaemic, antidiarrhoeal, antimicrobial, diuretic, antipyretic, psychopharmacological activities. To establish a simple, sensitive, reliable, rapid and validated high-performance thin-layer chromatography method for estimation of betulinic acid in hydro-alcoholic extract of N. nucifera Gaertn. rhizome. The separation was carried out on a thin-layer chromatography aluminium plate pre-coated with silica gel 60F(254) , eluted with chloroform, methanol and formic acid (49 : 1 : 1 v/v). Post chromatographic derivatisation was done with anisaldehyde-sulphuric acid reagent and densitometric scanning was performed using a Camag TLC scanner III, at 420 nm. The system was found to produce a compact spot for betulinic acid (R(f) = 0.30). A good linear precision relationship between the concentrations (2-10 µg) and peak areas were obtained with the correlation coefficient (r) of 0.99698. The limit of detection and limit of quantification of betulinic acid were detected to be 0.4 and 2.30 µg per spot. The percentage of recovery was found to be 98.36%. The percentage relative standard deviations of intra-day and inter-day precisions were 0.82-0.394 and 0.85-0.341, respectively. This validated HPTLC method provides a new and powerful approach to estimate betulinic acid as phytomarker in the extract. Copyright © 2010 John Wiley & Sons, Ltd.

  15. A rapid method for assessing the environmental performance of commercial farms in the Pampas of Argentina.

    Science.gov (United States)

    Viglizzo, E F; Frank, F; Bernardos, J; Buschiazzo, D E; Cabo, S

    2006-06-01

    The generation of reliable updated information is critical to support the harmonization of socio-economic and environmental issues in a context of sustainable development. The agro-environmental assessment and management of agricultural systems often relies on indicators that are necessary to make sound decisions. This work aims to provide an approach to (a) assess the environmental performance of commercial farms in the Pampas of Argentina, and (b) propose a methodological framework to calculate environmental indicators that can rapidly be applied to practical farming. 120 commercial farms scattered across the Pampas were analyzed in this study during 2002 and 2003. Eleven basic indicators were identified and calculation methods described. Such indicators were fossil energy (FE) use, FE use efficiency, nitrogen (N) balance, phosphorus (P) balance, N contamination risk, P contamination risk, pesticide contamination risk, soil erosion risk, habitat intervention, changes in soil carbon stock, and balance of greenhouse gases. A model named Agro-Eco-Index was developed on a Microsoft-Excel support to incorporate on-farm collected data and facilitate the calculation of indicators by users. Different procedures were applied to validate the model and present the results to the users. Regression models (based on linear and non-linear models) were used to validate the comparative performance of the study farms across the Pampas. An environmental dashboard was provided to represent in a graphical way the behavior of farms. The method provides a tool to discriminate environmentally friendly farms from those that do not pay enough attention to environmental issues. Our procedure might be useful for implementing an ecological certification system to reward a good environmental behavior in society (e.g., through tax benefits) and generate a commercial advantage (e.g., through the allocation of green labels) for committed farmers.

  16. A novel method for rapidly isolating microbes that suppress soil-borne phytopathogens

    Science.gov (United States)

    Cooper, Sarah; Agnew, Linda; Pereg, Lily

    2016-04-01

    Seedling establishment faces a large number of challenges related to soil physical properties as well as to fungal root diseases. It is extremely difficult to eliminate fungal pathogens from soils where their populations are established due to the persistent nature of their spores and since fumigation of resident fungi is very ineffective in clay-containing soils. Therefore it is necessary to find ways to overcome disease in areas where the soils are infected with fungal phytopathogens. The phenomenon of disease suppressive soils, where the pathogen is present but no disease observed, suggests that microbial antagonism in the soil may lead to the suppression of the growth of fungal pathogens. There are also cases in the literature where soil microorganisms were isolated that suppress the growth of phytopathogens. Antibiosis is one of the most important mechanisms responsible for fungal antagonism, with some significant antifungal compounds involved including antibiotics, volatile organic compounds, hydrogen cyanide and lytic enzymes. Isolation of pathogen-suppressive microorganisms from the soil is time consuming and tedious. We established a simple method for direct isolation of soil microbes (bacteria and fungi) that suppress fungal phytopathogens as well as procedures for confirmation of disease suppression. We will discuss such methods, which were so far tested with the cotton fungal pathogens Thielaviopsis basicola, Verticillium dahliae and Fusarium oxysporum and Verticillium fungicola. We have isolated a diversity of T. basicola-suppressive fungi and bacteria from two vastly different soil types. Identification of the antagonistic isolates revealed that they are a diverse lot, some belong to groups known to be suppressive of a wide range of fungal pathogens, endorsing the power of this technique to rapidly and directly isolate soil-borne microbes antagonistic to a wide variety of fungal pathogens.

  17. A simple and rapid flow cytometric method for detection of porcine cell surface markers.

    Science.gov (United States)

    Stabel, T J; Bolin, S R; Pesch, B A; Rahner, T E

    2000-11-01

    The objective of this study was to develop a rapid and reliable method for flow cytometric analysis of porcine whole blood cells. Fifty-microliters of heparin- or EDTA-treated whole blood was added to wells of a round-bottom 96-well microtitration plate. Each well contained 10 microl of an appropriate dilution of four different antibodies (40 microl total; two primary monoclonal antibodies and two fluorescent-labeled secondary antibodies). For convenience, the antibody mixture could be added to plates 1-2 days prior to assay and stored at 4 degrees C. Once whole blood was added to wells, plates were mixed gently, placed in a sealed bag and incubated in the dark at room temperature for 20 min. Contents of wells were then transferred to polystyrene tubes containing 2 ml of 1.5% formalin in distilled water and mixed gently. Cells were fixed for a minimum of 30 min and then stored in the dark at 4 degrees C until analysis by flow cytometry. Analysis of cell samples may be done up to 3 days after fixation. Results indicate that the percentages of Class I, Class II, CD3, CD8, CD4, CD45, monocyte, gamma-delta T-cell populations, and total number of granulocytes identified using this method were comparable to standard values or to values obtained following separation of white blood cells from red blood cells. The percentage of labeled B-cells was lower than standard values. Total assay time from receipt of blood to acquisition of data by flow cytometry required less than 2 h. This modified assay was shown to be simple, reliable, and useful for screening large numbers of porcine samples in a minimal period of time.

  18. A rapid method for detection of five known mutations associated with aminoglycoside-induced deafness.

    Science.gov (United States)

    Bardien, Soraya; Human, Hannique; Harris, Tashneem; Hefke, Gwynneth; Veikondis, Rene; Schaaf, H Simon; van der Merwe, Lize; Greinwald, John H; Fagan, Johan; de Jong, Greetje

    2009-01-13

    South Africa has one of the highest incidences of multidrug-resistant tuberculosis (MDR-TB) in the world. Concomitantly, aminoglycosides are commonly used in this country as a treatment against MDR-TB. To date, at least five mutations are known to confer susceptibility to aminoglycoside-induced hearing loss. The aim of the present study was to develop a rapid screening method to determine whether these mutations are present in the South African population. A multiplex method using the SNaPshot technique was used to screen for five mutations in the MT-RNR1 gene: A1555G, C1494T, T1095C, 961delT+C(n) and A827G. A total of 204 South African control samples, comprising 98 Mixed ancestry and 106 Black individuals were screened for the presence of the five mutations. A robust, cost-effective method was developed that detected the presence of all five sequence variants simultaneously. In this pilot study, the A1555G mutation was identified at a frequency of 0.9% in the Black control samples. The 961delT+C(n) variant was present in 6.6% of the Black controls and 2% of the Mixed ancestry controls. The T1095C, C1494T and A827G variants were not identified in any of the study participants. The frequency of 0.9% for the A1555G mutation in the Black population in South Africa is of concern given the high incidence of MDR-TB in this particular ethnic group. Future larger studies are warranted to determine the true frequencies of the aminoglycoside deafness mutations in the general South African population. The high frequencies of the 961delT+C(n) variant observed in the controls suggest that this change is a common non-pathogenic polymorphism. This genetic method facilitates the identification of individuals at high risk of developing hearing loss prior to the start of aminoglycoside therapy. This is important in a low-resource country like South Africa where, despite their adverse side-effects, aminoglycosides will continue to be used routinely and are accompanied with very

  19. ADVANTAGES OF RAPID METHOD FOR DETERMINING SCALE MASS AND DECARBURIZED LAYER OF ROLLED COIL STEEL

    Directory of Open Access Journals (Sweden)

    E. V. Parusov

    2016-08-01

    Full Text Available Purpose. To determine the universal empirical relationships that allow for operational calculation of scale mass and decarbonized layer depth based on the parameters of the technological process for rolled coil steel production. Methodology. The research is carried out on the industrial batches of the rolled steel of SAE 1006 and SAE 1065 grades. Scale removability was determined in accordance with the procedure of «Bekaert» company by the specifi-cations: GA-03-16, GA-03-18, GS-03-02, GS-06-01. The depth of decarbonized layer was identified in accordance with GOST 1763-68 (M method. Findings. Analysis of experimental data allowed us to determine the rational temperature of coil formation of the investigated steel grades, which provide the best possible removal of scale from the metal surface, a minimal amount of scale, as well as compliance of the metal surface color with the require-ments of European consumers. Originality. The work allowed establishing correlation of the basic quality indicators of the rolled coil high carbon steel (scale mass, depth of decarbonized layer and inter-laminar distance in pearlite with one of the main parameters (coil formation temperature of the deformation and heat treatment mode. The re-sulting regression equations, without metallographic analysis, can be used to determine, with a minimum error, the quantitative values of the total scale mass, depth of decarbonized layer and the average inter-lamellar distance in pearlite of the rolled coil high carbon steel. Practical value. Based on the specifications of «Bekaert» company (GA-03-16, GA-03-18, GS-03-02 and GS-06-01 the method of testing descaling by mechanical means from the surface of the rolled coil steel of low- and high-carbon steel grades was developed and approved in the environment of PJSC «ArcelorMittal Kryvyi Rih». The work resulted in development of the rapid method for determination of total and remaining scale mass on the rolled coil steel

  20. Development a rapid and accurate multiplex real time PCR method for the detection Chlamydia trachomatis and Mycoplasma hominis.

    Science.gov (United States)

    Safarkar, Roya; Mehrabadi, Jalil Fallah; Noormohammadi, Zahra; Mirnejad, Reza

    2017-11-01

    Sexually transmitted diseases easily spread among sexually active people and often have no symptoms. Rapid and accurate method for detecting these infections are necessary in early stages. The traditional detection methods of them are difficult and time-consuming. In this study, multiplex real time PCR was optimized for rapid identification of Chlamydia trachomatis and Mycoplasma hominis in a single tube and was performed with our designed primers. The sensitivity test was carried out to designed primers with diluted genomic DNA. To defined the specificity, non STD bacteria were used as DNA template. This study indicated that the developed multiplex real time PCR can be an effective alternative procedure to the conventional methods for rapid and accurate identification of C Chlamydia trachomatis and Mycoplasma hominis. Multiplex real-time PCR Results of them were checked with melting curves. The sensitivity of our designed primer by multiplex real time PCR for Chlamydia trachomatis and Mycoplasma hominis were 4.78×1010 and 8.35×1010 , respectively, Which the primers did not amplify any product from a non-STD species. Multiplex real time PCR by our new primers and analysis of melting curves were successfully usable for rapid and accurate detection of Chlamydia trachomatis and Mycoplasma hominis. This assay instead of traditional culture method, has considerable potential to be rapid, accurate and highly sensitive molecular diagnostic tool for simultaneous and direct detection. © 2017 Wiley Periodicals, Inc.

  1. Development of Novel Method for Rapid Extract of Radionuclides from Solution Using Polymer Ligand Film

    Science.gov (United States)

    Rim, Jung H.

    Accurate and fast determination of the activity of radionuclides in a sample is critical for nuclear forensics and emergency response. Radioanalytical techniques are well established for radionuclides measurement, however, they are slow and labor intensive, requiring extensive radiochemical separations and purification prior to analysis. With these limitations of current methods, there is great interest for a new technique to rapidly process samples. This dissertation describes a new analyte extraction medium called Polymer Ligand Film (PLF) developed to rapidly extract radionuclides. Polymer Ligand Film is a polymer medium with ligands incorporated in its matrix that selectively and rapidly extract analytes from a solution. The main focus of the new technique is to shorten and simplify the procedure necessary to chemically isolate radionuclides for determination by alpha spectrometry or beta counting. Five different ligands were tested for plutonium extraction: bis(2-ethylhexyl) methanediphosphonic acid (H2DEH[MDP]), di(2-ethyl hexyl) phosphoric acid (HDEHP), trialkyl methylammonium chloride (Aliquat-336), 4,4'(5')-di-t-butylcyclohexano 18-crown-6 (DtBuCH18C6), and 2-ethylhexyl 2-ethylhexylphosphonic acid (HEH[EHP]). The ligands that were effective for plutonium extraction further studied for uranium extraction. The plutonium recovery by PLFs has shown dependency on nitric acid concentration and ligand to total mass ratio. H2DEH[MDP] PLFs performed best with 1:10 and 1:20 ratio PLFs. 50.44% and 47.61% of plutonium were extracted on the surface of PLFs with 1M nitric acid for 1:10 and 1:20 PLF, respectively. HDEHP PLF provided the best combination of alpha spectroscopy resolution and plutonium recovery with 1:5 PLF when used with 0.1M nitric acid. The overall analyte recovery was lower than electrodeposited samples, which typically has recovery above 80%. However, PLF is designed to be a rapid field deployable screening technique and consistency is more important

  2. Rapid detection of coliforms in drinking water of Arak city using multiplex PCR method in comparison with the standard method of culture (Most Probably Number)

    OpenAIRE

    fatemeh, Dehghan; Reza, Zolfaghari Mohammad; Mohammad, Arjomandzadegan; Salomeh, Kalantari; Reza, Ahmari Gholam; Hossein, Sarmadian; Maryam, Sadrnia; Azam, Ahmadi; Mana, Shojapoor; Negin, Najarian; Reza, Kasravi Alii; Saeed, Falahat

    2014-01-01

    Objective: To analyse molecular detection of coliforms and shorten the time of PCR. Methods: Rapid detection of coliforms by amplification of lacZ and uidA genes in a multiplex PCR reaction was designed and performed in comparison with most probably number (MPN) method for 16 artificial and 101 field samples. The molecular method was also conducted on isolated coliforms from positive MPN samples; standard sample for verification of microbial method certificated reference material; isolated...

  3. A rapid method for an offline glycerol determination during microbial fermentation

    Directory of Open Access Journals (Sweden)

    Jennifer Kuhn

    2015-05-01

    Conclusions: With this rapid assay, glycerol could be detected easily in microbial fermentation broth. It is reliable over a wide concentration range including advantages such as an easy assay set-up, a short assay time and no sample pretreatment.

  4. Problems with rapid agglutination methods for identification of Staphylococcus aureus when Staphylococcus saprophyticus is being tested.

    OpenAIRE

    Gregson, D B; Low, D E; Skulnick, M; Simor, A E

    1988-01-01

    Six rapid agglutination tests for identification of Staphylococcus aureus were evaluated by using 62 strains of S. aureus, 63 strains of S. saprophyticus, and 67 strains of other coagulase-negative staphylococci. S. saprophyticus was responsible for 19 of 26 false-positive results and 20 uninterpretable reactions. Thus, urinary staphylococcal isolates that are positive by rapid agglutination tests may require other confirmatory tests for the identification of possible S. saprophyticus.

  5. A Rapid and Reliable Method for Total Protein Extraction from Succulent Plants for Proteomic Analysis.

    Science.gov (United States)

    Lledías, Fernando; Hernández, Felipe; Rivas, Viridiana; García-Mendoza, Abisaí; Cassab, Gladys I; Nieto-Sotelo, Jorge

    2017-08-01

    Crassulacean acid metabolism plants have some morphological features, such as succulent and reduced leaves, thick cuticles, and sunken stomata that help them prevent excessive water loss and irradiation. As molecular constituents of these morphological adaptations to xeric environments, succulent plants produce a set of specific compounds such as complex polysaccharides, pigments, waxes, and terpenoids, to name a few, in addition to uncharacterized proteases. Since all these compounds interfere with the analysis of proteins by electrophoretic techniques, preparation of high quality samples from these sources represents a real challenge. The absence of adequate protocols for protein extraction has restrained the study of this class of plants at the molecular level. Here, we present a rapid and reliable protocol that could be accomplished in 1 h and applied to a broad range of plants with reproducible results. We were able to obtain well-resolved SDS/PAGE protein patterns in extracts from different members of the subfamilies Agavoideae (Agave, Yucca, Manfreda, and Furcraea), Nolinoideae (Dasylirion and Beucarnea), and the Cactaceae family. This method is based on the differential solubility of contaminants and proteins in the presence of acetone and pH-altered solutions. We speculate about the role of saponins and high molecular weight carbohydrates to produce electrophoretic-compatible samples. A modification of the basic protocol allowed the analysis of samples by bidimensional electrophoresis (2DE) for proteomic analysis. Furostanol glycoside 26-O-β-glucosidase (an enzyme involved in steroid saponin synthesis) was successfully identified by mass spectrometry analysis and de novo sequencing of a 2DE spot from an Agave attenuata sample.

  6. Biogas production from ensiled meadow grass; effect of mechanical pretreatments and rapid determination of substrate biodegradability via physicochemical methods

    DEFF Research Database (Denmark)

    Tsapekos, Panagiotis; Kougias, Panagiotis; Angelidaki, Irini

    2015-01-01

    As the biogas sector is rapidly expanding, there is an increasing need in finding new alternative feedstock to biogas plants. Meadow grass can be a suitable co-substrate and if ensiled it can be supplied to biogas plants continuously throughout the year. Nevertheless, this substrate is quite reca...... the methods, electrical conductivity test showed the most promising calibration statistics (R2 = 0.68)....... increased the methane productivity and the increase ranged from 8% to 25%. The best mechanical pretreatment was the usage of two coarse mesh grating plates. Additionally, simple analytical methods were conducted to investigate the possibility of rapidly determining the methane yield of meadow grass. Among...

  7. A rapid and quantitative method to detect human circulating tumor cells in a preclinical animal model.

    Science.gov (United States)

    Tu, Shih-Hsin; Hsieh, Yi-Chen; Huang, Li-Chi; Lin, Chun-Yu; Hsu, Kai-Wen; Hsieh, Wen-Shyang; Chi, Wei-Ming; Lee, Chia-Hwa

    2017-06-23

    As cancer metastasis is the deadliest aspect of cancer, causing 90% of human deaths, evaluating the molecular mechanisms underlying this process is the major interest to those in the drug development field. Both therapeutic target identification and proof-of-concept experimentation in anti-cancer drug development require appropriate animal models, such as xenograft tumor transplantation in transgenic and knockout mice. In the progression of cancer metastasis, circulating tumor cells (CTCs) are the most critical factor in determining the prognosis of cancer patients. Several studies have demonstrated that measuring CTC-specific markers in a clinical setting (e.g., flow cytometry) can provide a current status of cancer development in patients. However, this useful technique has rarely been applied in the real-time monitoring of CTCs in preclinical animal models. In this study, we designed a rapid and reliable detection method by combining a bioluminescent in vivo imaging system (IVIS) and quantitative polymerase chain reaction (QPCR)-based analysis to measure CTCs in animal blood. Using the IVIS Spectrum CT System with 3D-imaging on orthotropic-developed breast-tumor-bearing mice. In this manuscript, we established a quick and reliable method for measuring CTCs in a preclinical animal mode. The key to this technique is the use of specific human and mouse GUS primers on DNA/RNA of mouse peripheral blood under an absolute qPCR system. First, the high sensitivity of cancer cell detection on IVIS was presented by measuring the luciferase carried MDA-MB-231 cells from 5 to 5x10(11) cell numbers with great correlation (R(2) = 0.999). Next, the MDA-MB-231 cell numbers injected by tail vein and their IVIS radiance signals were strongly corrected with qPCR-calculated copy numbers (R(2) > 0.99). Furthermore, by applying an orthotropic implantation animal model, we successfully distinguished xenograft tumor-bearing mice and control mice with a significant difference (p < 0

  8. Rapid Analytical Methods for On-Site Triage for Traumatic Brain Injury

    Science.gov (United States)

    North, Stella H.; Shriver-Lake, Lisa C.; Taitt, Chris R.; Ligler, Frances S.

    2012-07-01

    Traumatic brain injury (TBI) results from an event that causes rapid acceleration and deceleration of the brain or penetration of the skull with an object. Responses to stimuli and questions, loss of consciousness, and altered behavior are symptoms currently used to justify brain imaging for diagnosis and therapeutic guidance. Tests based on such symptoms are susceptible to false-positive and false-negative results due to stress, fatigue, and medications. Biochemical markers of neuronal damage and the physiological response to that damage are being identified. Biosensors capable of rapid measurement of such markers in the circulation offer a solution for on-site triage, as long as three criteria are met: (a) Recognition reagents can be identified that are sufficiently sensitive and specific, (b) the biosensor can provide quantitative assessment of multiple markers rapidly and simultaneously, and (c) both the sensor and reagents are designed for use outside the laboratory.

  9. A rapid sonication based method for preparation of stromal vascular fraction and mesenchymal stem cells from fat tissue

    Directory of Open Access Journals (Sweden)

    Mohammad Amir Amirkhani

    2016-06-01

    Conclusion: The current protocol based on the sonication-mediated cavitation is a rapid, safe and cost-effective method, which is proposed for isolation of SVF and of course ADSCs cultures in a large scale for the clinical trials or therapeutic purposes.

  10. A RAPID DNA EXTRACTION METHOD IS SUCCESSFULLY APPLIED TO ITS-RFLP ANALYSIS OF MYCORRHIZAL ROOT TIPS

    Science.gov (United States)

    A rapid method for extracting DNA from intact, single root tips using a Xanthine solution was developed to handle very large numbers of analyses of ectomycorrhizas. By using an extraction without grinding we have attempted to bias the extraction towards the fungal DNA in the man...

  11. A Boussinesq-type method for fully nonlinear waves interacting with a rapidly varying bathymetry

    DEFF Research Database (Denmark)

    Madsen, Per A.; Fuhrman, David R.; Wang, Benlong

    2006-01-01

    New equations are derived for fully nonlinear and highly dispersive water waves interacting with a rapidly varying bathymetry. The derivation is an extension of a recent high order Boussinesq type formulation valid on a mildly sloping bottom. It is based on a series expansion from a rapidly...... locally deteriorate, and we provide a guideline for using this technique within acceptable accuracy bounds. Numerical results are given for the linear reflection from a plane shelf, a Gaussian shaped trench, and a symmetric trench with sloped transitions. Furthermore, we simulate the linear class I...

  12. Comparing different methods for fast screening of microbiological quality of beach sand aimed at rapid-response remediation.

    Science.gov (United States)

    Testolin, Renan C; Almeida, Tito C M; Polette, Marcus; Branco, Joaquim O; Fischer, Larissa L; Niero, Guilherme; Poyer-Radetski, Gabriel; Silva, Valéria C; Somensi, Cleder A; Corrêa, Albertina X R; Corrêa, Rogério; Rörig, Leonardo R; Itokazu, Ana Gabriela; Férard, Jean-François; Cotelle, Sylvie; Radetski, Claudemir M

    2017-05-15

    There is scientific evidence that beach sands are a significant contributor to the pathogen load to which visitors are exposed. To develop beach quality guidelines all beach zones must be included in microbiological evaluations, but monitoring methods for beach sand quality are relatively longstanding, expensive, laborious and require moderate laboratory infrastructure. This paper aimed to evaluate the microorganism activity in different beach zones applying and comparing a classical method of membrane filtration (MF) with two colorimetric screening methods based on fluorescein (FDA) and tetrazolium (TTC) salt biotransformation to evaluate a new rapid and low-cost method for beach sand microbiological contamination assessments. The colorimetric results can help beach managers to evaluate rapidly and at low cost the microbiological quality of different beach zones in order to decide whether remedial actions need to be adopted to prevent exposure of the public to microbes due to beach sand and/or water contamination. Copyright © 2017. Published by Elsevier Ltd.

  13. 1 H-NMR metabolomics: Profiling method for a rapid and efficient ...

    African Journals Online (AJOL)

    Principal component analysis was used to separate groups of samples and to relate known and unknown metabolites to transgenic events. The screening of 100 samples, from extraction to data mining, took 36 h. Thus, this procedure allows the rapid selection of metabolic phenotypes of interest among about 30 transgenic ...

  14. Setaria viridis floral-dip: A simple and rapid Agrobacterium-medicated transformation method

    Science.gov (United States)

    Setaria viridis was recently described as a new monocotyledonous model species for C4 photosynthesis research and genetic transformation. It has biological attributes (rapid life cycle, small genome, diploid, short stature and simple growth requirements) that make it suitable for use as a model plan...

  15. Rapid HPLC method to screen pectins for heterogeneity in methyl-esterification and amidation

    NARCIS (Netherlands)

    Guillotin, S.E.; Loey, van A.; Boulenguer, P.; Schols, H.A.; Voragen, A.G.J.

    2007-01-01

    Functionality of pectins as a food ingredient is strongly related to their chemical fine structure. Chemical characteristics of pectins are determined by many different parameters in their manufacture (choice of the raw material and extraction conditions). Pectin companies are thus in need of rapid

  16. NMR analysis of budding yeast metabolomics: a rapid method for sample preparation.

    Science.gov (United States)

    Airoldi, C; Tripodi, F; Guzzi, C; Nicastro, R; Coccetti, P

    2015-02-01

    Here we propose the optimization of a rapid and reproducible protocol for intracellular metabolite extraction from yeast cells and their metabolic profiling by (1)H-NMR spectroscopy. The protocol reliability has been validated through comparison between the metabolome of cells in different phases of growth or with different genetic backgrounds.

  17. Collision-induced fragmentation accurate mass spectrometric analysis methods to rapidly characterize plant extracts

    Science.gov (United States)

    The rapid advances in analytical chromatography equipment have made the reliable and reproducible measurement of a wide range of plant chemical components possible. Full chemical characterization of a given plant material is possible with the new mass spectrometers currently available. For phytochem...

  18. Direct PCR - A rapid method for multiplexed detection of different serotypes of Salmonella in enriched pork meat samples

    DEFF Research Database (Denmark)

    Chin, Wai Hoe; Sun, Yi; Høgberg, Jonas

    2017-01-01

    , in this study, we developed a multiplex Direct PCR method for rapid detection of different Salmonella serotypes directly from pork meat samples without any DNA purification steps. An inhibitor-resistant Phusion Pfu DNA polymerase was used to overcome PCR inhibition. Four pairs of primers including a pair...... of newly designed primers targeting Salmonella spp. at subtype level were incorporated in the multiplex Direct PCR. To maximize the efficiency of the Direct PCR, the ratio between sample and dilution buffer was optimized. The sensitivity and specificity of the multiplex Direct PCR were tested using...... and integration into a point-of-need Lab-on-a-chip system for rapid online pathogen detection....

  19. A rapid method of detecting motor blocks in patients with Parkinson's disease during volitional hand movements

    Directory of Open Access Journals (Sweden)

    Popović Mirjana B.

    2002-01-01

    -5%; 56% had MBT% 5-10%; 22% had MBT% 10-15%; 5.5% had MBT% 15-20°% and 2% had MBT% 20-25%. No block lasted more than 25% from the whole movement duration. Table 2 is the summary of mean variability for kinematic indicators of motor block (N, mbt%, t% and for the movement duration T during a 7 day-testing of patients #3. The analysis of calculated data for eight tested PD patients revealed a significant difference (p < 0.01 between healthy controls and three PD patients; data on five PD patients were not significantly different (ns. This method clustered 3 PD patients in the group that experience motor blocks, while the rest were in the group without their significant occurrence. DISCUSSION This algorithm is an additional instrument in classical evaluation of PD patients during their clinical evaluation and treatment. It provides to clinician a rapid feedback on the changes of voluntary hand movements in everyday progress of illness. Furthermore, this method could be of assistance for developing strategies to overcome motor blocks in arm movements at their beginning, as well as for the feedback of the success of drug therapy.

  20. A rapid, efficient, and economic device and method for the isolation and purification of mouse islet cells.

    Directory of Open Access Journals (Sweden)

    Yin Zongyi

    Full Text Available Rapid, efficient, and economic method for the isolation and purification of islets has been pursued by numerous islet-related researchers. In this study, we compared the advantages and disadvantages of our developed patented method with those of commonly used conventional methods (Ficoll-400, 1077, and handpicking methods. Cell viability was assayed using Trypan blue, cell purity and yield were assayed using diphenylthiocarbazone, and islet function was assayed using acridine orange/ethidium bromide staining and enzyme-linked immunosorbent assay-glucose stimulation testing 4 days after cultivation. The results showed that our islet isolation and purification method required 12 ± 3 min, which was significantly shorter than the time required in Ficoll-400, 1077, and HPU groups (34 ± 3, 41 ± 4, and 30 ± 4 min, respectively; P 1000 islets. In summary, the MCT method is a rapid, efficient, and economic method for isolating and purifying murine islet cell clumps. This method overcomes some of the shortcomings of conventional methods, showing a relatively higher quality and yield of islets within a shorter duration at a lower cost. Therefore, the current method provides researchers with an alternative option for islet isolation and should be widely generalized.

  1. A rapid, efficient, and economic device and method for the isolation and purification of mouse islet cells.

    Science.gov (United States)

    Zongyi, Yin; Funian, Zou; Hao, Li; Ying, Cheng; Jialin, Zhang; Baifeng, Li

    2017-01-01

    Rapid, efficient, and economic method for the isolation and purification of islets has been pursued by numerous islet-related researchers. In this study, we compared the advantages and disadvantages of our developed patented method with those of commonly used conventional methods (Ficoll-400, 1077, and handpicking methods). Cell viability was assayed using Trypan blue, cell purity and yield were assayed using diphenylthiocarbazone, and islet function was assayed using acridine orange/ethidium bromide staining and enzyme-linked immunosorbent assay-glucose stimulation testing 4 days after cultivation. The results showed that our islet isolation and purification method required 12 ± 3 min, which was significantly shorter than the time required in Ficoll-400, 1077, and HPU groups (34 ± 3, 41 ± 4, and 30 ± 4 min, respectively; P 1000 islets). In summary, the MCT method is a rapid, efficient, and economic method for isolating and purifying murine islet cell clumps. This method overcomes some of the shortcomings of conventional methods, showing a relatively higher quality and yield of islets within a shorter duration at a lower cost. Therefore, the current method provides researchers with an alternative option for islet isolation and should be widely generalized.

  2. Passive acoustic methods for fine-scale tracking of harbour porpoises in tidal rapids

    DEFF Research Database (Denmark)

    Macaulay, Jamie; Gordon, Jonathan; Gillespie, Douglas

    2017-01-01

    The growing interest in generating electrical power from tidal currents using tidal turbine generators raises a number of environmental concerns, including the risk that marine mammals might be injured or killed through collision with rotating turbine blades. To understand this risk, information...... on how marine mammals use tidal rapid habitats and in particular, their underwater movements and dive behaviour is required. Porpoises, which are the most abundant small cetacean at most European tidal sites, are difficult animals to tag, and the limited size of tidal habitats means that any telemetered...... animal would be likely to spend only a small proportion of time within them. Here, an alternative approach is explored, whereby passive acoustic monitoring (PAM) is used to obtain fine scale geo-referenced tracks of harbour porpoises in tidal rapid areas. Large aperture hydrophone arrays are required...

  3. Development of rapid methods for relaxation time mapping and motion estimation using magnetic resonance imaging

    OpenAIRE

    Gilani, Syed Irtiza Ali

    2008-01-01

    Recent technological developments in the field of magnetic resonance imaging have resulted in advanced techniques that can reduce the total time to acquire images. For applications such as relaxation time mapping, which enables improved visualisation of in vivo structures, rapid imaging techniques are highly desirable. TAPIR is a Look- Locker-based sequence for high-resolution, multislice T1 relaxation time mapping. Despite the high accuracy and precision of TAPIR, an improveme...

  4. A facile and rapid method for the black pepper leaf mediated green synthesis of silver nanoparticles and the antimicrobial study

    Science.gov (United States)

    Augustine, Robin; Kalarikkal, Nandakumar; Thomas, Sabu

    2014-10-01

    Green synthesis of nanoparticles is widely accepted due to the less toxicity in comparison with chemical methods. But there are certain drawbacks like slow formation of nanoparticles, difficulty to control particle size and shape make them less convenient. Here we report a novel cost-effective and eco-friendly method for the rapid green synthesis of silver nanoparticles using leaf extracts of Piper nigrum. Our results suggest that this method can be used for obtaining silver nanoparticles with controllable size within a few minutes. The fabricated nanoparticles possessed excellent antibacterial property against both Gram-positive and Gram-negative bacteria.

  5. DPS – A rapid method for genome sequencing of DNA-containing bacteriophages directly from a single plaque

    DEFF Research Database (Denmark)

    Kot, Witold; Vogensen, Finn Kvist; Sørensen, Søren J.

    2014-01-01

    methods for characterization of phages is determination of the whole genome using high throughput sequencing approaches. Here a direct plaque sequencing (DPS) is described, which is a rapid method that allows easy full genome sequencing of DNA-containing phages using the Nextera XT™ kit. A combination...... of host-DNA removal followed by purification and concentration of the viral DNA, allowed the construction of Illumina-compatible sequencing libraries using the Nextera™ XT technology directly from single phage plaques without any whole genome amplification step. This method was tested on three...

  6. Rapid Solid-Liquid Dynamic Extraction (RSLDE): a New Rapid and Greener Method for Extracting Two Steviol Glycosides (Stevioside and Rebaudioside A) from Stevia Leaves.

    Science.gov (United States)

    Gallo, Monica; Vitulano, Manuela; Andolfi, Anna; DellaGreca, Marina; Conte, Esterina; Ciaravolo, Martina; Naviglio, Daniele

    2017-06-01

    Stevioside and rebaudioside A are the main diterpene glycosides present in the leaves of the Stevia rebaudiana plant, which is used in the production of foods and low-calorie beverages. The difficulties associated with their extraction and purification are currently a problem for the food processing industries. The objective of this study was to develop an effective and economically viable method to obtain a high-quality product while trying to overcome the disadvantages derived from the conventional transformation processes. For this reason, extractions were carried out using a conventional maceration (CM) and a cyclically pressurized extraction known as rapid solid-liquid dynamic extraction (RSLDE) by the Naviglio extractor (NE). After only 20 min of extraction using the NE, a quantity of rebaudioside A and stevioside equal to 1197.8 and 413.6 mg/L was obtained, respectively, while for the CM, the optimum time was 90 min. From the results, it can be stated that the extraction process by NE and its subsequent purification developed in this study is a simple, economical, environmentally friendly method for producing steviol glycosides. Therefore, this method constitutes a valid alternative to conventional extraction by reducing the extraction time and the consumption of toxic solvents and favouring the use of the extracted metabolites as food additives and/or nutraceuticals. As an added value and of local interest, the experiment was carried out on stevia leaves from the Benevento area (Italy), where a high content of rebaudioside A was observed, which exhibits a sweet taste compared to stevioside, which has a significant bitter aftertaste.

  7. Rapid concentration and sensitive detection of hookworm ova from wastewater matrices using a real-time PCR method.

    Science.gov (United States)

    Gyawali, P; Sidhu, J P S; Ahmed, W; Jagals, P; Toze, S

    2015-12-01

    The risk of human hookworm infections from land application of wastewater matrices could be high in regions with high hookworm prevalence. A rapid, sensitive and specific hookworm detection method from wastewater matrices is required in order to assess human health risks. Currently available methods used to identify hookworm ova to the species level are time consuming and lack accuracy. In this study, a real-time PCR method was developed for the rapid, sensitive and specific detection of canine hookworm (Ancylostoma caninum) ova from wastewater matrices. A. caninum was chosen because of its morphological similarity to the human hookworm (Ancylostoma duodenale and Necator americanus). The newly developed PCR method has high detection sensitivity with the ability to detect less than one A. caninum ova from 1 L of secondary treated wastewater at the mean threshold cycle (CT) values ranging from 30.1 to 34.3. The method is also able to detect four A. caninum ova from 1 L of raw wastewater and from ∼4 g of treated sludge with mean CT values ranging from 35.6 to 39.8 and 39.8 to 39.9, respectively. The better detection sensitivity obtained for secondary treated wastewater compared to raw wastewater and sludge samples could be attributed to sample turbidity. The proposed method appears to be rapid, sensitive and specific compared to traditional methods and has potential to aid in the public health risk assessment associated with land application of wastewater matrices. Furthermore, the method can be adapted to detect other helminth ova of interest from wastewater matrices. Crown Copyright © 2015. Published by Elsevier Inc. All rights reserved.

  8. Rapid determination method for 2-DCB in irradiated pork by ASE-Al2O3-GC-MS

    Science.gov (United States)

    Li, An; Pan, Li-Gang; Zhang, Xiu-Tong; Jin, Xin-Xin; Ma, Hong-Zao

    2017-01-01

    2-Dodecylcyclobutanone (2-DCB) is validated as a marker for irradiated foodstuffs. However, the preparation methods for the marker compound involve time-consuming and tedious procedures prior to analysis. This paper describes a rapid method for the extraction and cleanup of 2-DCB in irradiated pork. The highly automated extraction procedure is based on accelerated solvent extraction combined with a purification step by Al2O3, which is added to the extraction cell. The overall extraction time is less than 20 min, and the volume of the solvent is 1.5 times that of the extraction cell (34 mL). The recovery of 2-DCB from pork samples for this method is higher than 75%. The method allows detecting pork irradiated at low doses (0.5 kGy). The proposed method is considered an alternative to the Soxhlet extraction and Florisil chromatography method currently in use.

  9. A simple and rapid visual method for the determination of ammonia nitrogen in environmental waters using thymol

    Energy Technology Data Exchange (ETDEWEB)

    Okumura, M.; Fujinaga, K.; Seike, Y.; Honda, S. [Dept. of Material Science, Interdisciplinary Faculty of Science and Engineering, Shimane University, Matsue (Japan)

    1999-11-01

    Simple visual and spectrophotometric methods for the determination of ammonia nitrogen in water are proposed, based on the color development of indothymol blue formed between ammonia and thymol. The color development was accelerated by nitroprusside to complete in 3 min. This color development is remarkably rapid compared with that of the other conventional methods with indothymol blue and indophenol blue. The concentration range of ammonia nitrogen spectrophotometrically determined was 0.04-1.2 mg/L NH{sub 4}-N. The absorbance per 1 {mu}g NH{sub 4}-N was 0.0215 (molar absorptivity = 1.51 x 10{sup 4}) at 690 nm. The visual method not using any instrument as an in situ method in field works was developed based on the optimum conditions for the established spectrophotometric method. This visual method was successfully applied to the determination of ammonia nitrogen in environmental waters. (orig.)

  10. A novel method for perceptual assessment of small room acoustics using rapid sensory analysis

    DEFF Research Database (Denmark)

    Kaplanis, Neofytos; Bech, Søren; Lokki, Tapio

    2016-01-01

    small rooms and car cabins. In-situ measurements were performed to obtain a range of possible acoustical settings, by varying physically the spaces under investigation. The measured responses were spatiallyanalyzed and synthesized to reproduce the observed fields in the laboratory. Expert listeners were...... presented with auralized sound over a loudspeaker array and followed a rapid sensory analysis protocol. The elicited attributes and ratings are analyzed and possible links to the acoustical properties of these spaces are discussed. [This study is a part of Marie Curie Network on Dereverberation...

  11. Rapid prototyping tools and methods for all-Topas (R) cyclic olefin copolymer fluidic microsystems

    DEFF Research Database (Denmark)

    Bundgaard, Frederik; Perozziello, Gerardo; Geschke, Oliver

    2006-01-01

    Topas (R), the cyclic olefin copolymer, from Topas Advanced Polymers GmbH has a number of advantages over polymers such as poly(methylmethacrylate), polydimethylsiloxane, and polycarbonate traditionally used in fluid microsystem manufacturing, such as low water absorption, high chemical resistance......, good machinability, and good optical properties. A number of different processes for rapid and low-cost prototyping of all-Topas microfluidic systems, made with desktop machinery, are presented. Among the processes are micromilling of fluidic structures with a width down to 25 p,m and sealing...

  12. MacroEvoLution: A New Method for the Rapid Generation of Novel Scaffold-Diverse Macrocyclic Libraries.

    Science.gov (United States)

    Saupe, Jörn; Kunz, Oliver; Haustedt, Lars Ole; Jakupovic, Sven; Mang, Christian

    2017-09-04

    Macrocycles are a structural class bearing great promise for future challenges in medicinal chemistry. Nevertheless, there are few flexible approaches for the rapid generation of structurally diverse macrocyclic compound collections. Here, an efficient method for the generation of novel macrocyclic peptide-based scaffolds is reported. The process, named here as "MacroEvoLution", is based on a cyclization screening approach that gives reliable access to novel macrocyclic architectures. Classification of building blocks into specific pools ensures that scaffolds with orthogonally addressable functionalities are generated, which can easily be used for the generation of structurally diverse compound libraries. The method grants rapid access to novel scaffolds with scalable synthesis (multi gram scale) and the introduction of further diversity at a late stage. Despite being developed for peptidic systems, the approach can easily be extended for the synthesis of systems with a decreased peptidic character. © 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

  13. MacroEvoLution: A New Method for the Rapid Generation of Novel Scaffold‐Diverse Macrocyclic Libraries

    Science.gov (United States)

    Saupe, Jörn; Kunz, Oliver; Haustedt, Lars Ole; Jakupovic, Sven

    2017-01-01

    Abstract Macrocycles are a structural class bearing great promise for future challenges in medicinal chemistry. Nevertheless, there are few flexible approaches for the rapid generation of structurally diverse macrocyclic compound collections. Here, an efficient method for the generation of novel macrocyclic peptide‐based scaffolds is reported. The process, named here as “MacroEvoLution”, is based on a cyclization screening approach that gives reliable access to novel macrocyclic architectures. Classification of building blocks into specific pools ensures that scaffolds with orthogonally addressable functionalities are generated, which can easily be used for the generation of structurally diverse compound libraries. The method grants rapid access to novel scaffolds with scalable synthesis (multi gram scale) and the introduction of further diversity at a late stage. Despite being developed for peptidic systems, the approach can easily be extended for the synthesis of systems with a decreased peptidic character. PMID:28715083

  14. Evaluation of culture methods for rapid screening of swine faecal samples for Yersinia enterocolitica O : 3 biotype 4

    DEFF Research Database (Denmark)

    Hoorfar, Jeffrey; Holmvig, C.B.F.

    1999-01-01

    In two studies, seven different culture protocols were compared to test naturally contaminated faecal samples from pigs for isolation of Y. enterocolitica serotype O; 3/biotype 4( n = 70 and n = 79). Four of the protocols were based on the Nordic Committee on Food Analysis (NMKL protocols), while...... three protocols were based on a rapid and selective method (here called ITC protocols). The protocols differed mainly in time of pre-enrichment (1, 10 and 24 d) and enrichment (2, 10, 24 d) and the type of selective enrichment media (ITC vs. MRB). The sensitivity of the rapid ITC protocol (24% and 9...... indicate possibilities of shortening the culture methods by replacing most of the biochemical tests with an agglutination test based on a monoclonal antibody....

  15. Designing and comparison study of rapid detection methods of resistance to injectable drugs in clinical strains of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Fatemeh Salehi

    2012-01-01

    Full Text Available Introduction: In this study, some molecular methods were designed for rapid detection of resistance to kanamycin and amikacin.Materials and methods: Among 120 clinical isolates of mycobacterium tuberculosis, 70 strains were selected for evaluation of possible mutations. A PCR-RFLP method was designed for detection of wild type (using enzyme ajii and mutant from (BstFNI enzyme of the isolates. Furthermore, allele specific method (as PCR was designed for detection mutations in codons 1401 and 1402 gene rrs. Some selected isolates were sequenced.Results: In PCR-RFLP method, among the 70 strains examined by BstFNI enzyme, could detect 17 mutant strains among 24 phenotypicaly resistant and 44 non-mutant isolates from 46 susceptible isolates. The sensitivity of this method was %70.83 and specificity was %95.65 on the other hand, 12 mutant from 20 resistant strains and 29 non-mutant strains from 32 susceptible strains were detected by AjiI enzyme. The sensitivity and specificity of this method was 60 and %90.62, respectively. In MAS PCR, 3 mutants from 6 resistant strains and 12 non-mutants from 17 resistant strains were detected. The sensitivity of this method was 50 and specificity was 70.58. Results of sequencing method confirmed the results of molecular methods.Discussion and conclusion: PCR-RFLP method by BstFNI enzyme was the best method for rapid detection of Mycobacterium tuberculosis resistant to second-line injectable drugs and was recommended for routine use.

  16. rapid mini-prep DNA extraction method in rice (Oryza sativa ...

    African Journals Online (AJOL)

    method, optimized for rice, which was achieved via some modifications in present DNA extraction methods, especially in first step of cell wall lyses and the use of cheap and frequent chemicals found in every laboratory is presented. Normal quality and quantity was obtained by the method. The PCR based assays also ...

  17. [A rapid method for the quantitative determination of protein in urine].

    Science.gov (United States)

    Kariagina, I Iu; Slepysheva, V V; Kozlov, A V

    1996-01-01

    A simple, economic, and available method for measuring protein in the urine is proposed. It is based on the capacity of bromophenol blue stain to form a complex with proteins in acid medium, which is characterized by absorption maximum at a wave-length of 597 nm. The proposed method is compared with the sulfosalicylic acid test and the Biuret method.

  18. A rapid and low-cost DNA extraction method for isolating ...

    African Journals Online (AJOL)

    The price of commercial DNA extraction methods makes the routine use of polymerase chain reaction amplification (PCR) based methods rather costly for scientists in developing countries. A guanidium thiocayante-based DNA extraction method was investigated in this study for the isolation of Escherichia coli (E. coli) DNA ...

  19. Exploration and practice of methods and processes of evidence-based rapid review on peer review of WHO EML application.

    Science.gov (United States)

    Li, Youping; Yu, Jiajie; Du, Liang; Sun, Xin; Kwong, Joey S W; Wu, Bin; Hu, Zhiqiang; Lu, Jing; Xu, Ting; Zhang, Lingli

    2015-11-01

    After 38 years of development, the procedure of selection and evaluation of the World Health Organization Essential Medicine List (WHO EML) is increasingly scientific and formal. However, peer review for the applications of World Health Organization Essential Medicine List is always required in a short period. It is necessary to build up a set of methods and processes for rapid review. We identified the process of evidenced-based rapid review on WHO EML application for peer reviews according to 11 items which were required during reporting of the peer review results of the proposals. The most important items for the rapid review of World Health Organization Essential Medicine List peer reviewers are (1) to confirm the requirements and identify the purposes; (2) to establish the research questions and translate the questions into the 'Participants, Interventions, Comparators, Outcomes, Study design' (PICOS) format; (3) to search and screen available evidence, for which high-level evidence is preferred, such as systematic reviews or meta-analyses, health technology assessment, clinical guidelines; (4) to extract data, where we extract primary information based on the purposes; (5) to synthesize data by qualitative methods, assess the quality of evidence, and compare the results; (6) to provide the answers to the applications, quality of evidences and strength of recommendations. Our study established a set of methods and processes for the rapid review of World Health Organization Essential Medicine List peer review, and our findings were used to guide the reviewers to fulfill the 19(th) World Health Organization Essential Medicine List peer review. The methods and processes were feasible and met the necessary requirements in terms of time and quality. Continuous improvement and evaluation in practice are warranted. © 2015 Chinese Cochrane Center, West China Hospital of Sichuan University and Wiley Publishing Asia Pty Ltd.

  20. Novel, rapid optical immunoassay technique for detection of group A streptococci from pharyngeal specimens: comparison with standard culture methods.

    OpenAIRE

    Harbeck, R. J.; Teague, J; Crossen, G R; Maul, D M; Childers, P L

    1993-01-01

    A novel immunoassay system based on the changes in the reflection of light, termed an optical immunoassay (OIA), was utilized to directly detect group A streptococcal (GAS) carbohydrate antigen from clinical specimens. In two studies, a total of 1,275 throat swabs were tested for the presence of this antigen with the Strep A OIA rapid detection system and the results were compared with those of standard culture methods. In both studies, the Strep A OIA yielded more positive results than plati...

  1. Rapid and Sensitive Lateral Flow Immunoassay Method for Procalcitonin (PCT) Based on Time-Resolved Immunochromatography.

    Science.gov (United States)

    Shao, Xiang-Yang; Wang, Cong-Rong; Xie, Chun-Mei; Wang, Xian-Guo; Liang, Rong-Liang; Xu, Wei-Wen

    2017-02-28

    Procalcitonin (PCT) is a current, frequently-used marker for severe bacterial infection. The aim of this study was to develop a cost-effective detection kit for rapid quantitative and on-site detection of PCT. To develop the new PCT quantitative detecting kit, a double-antibody sandwich immunofluorescent assay was employed based on time-resolved immunofluorescent assay (TRFIA) combined with lateral flow immunoassay (LFIA). The performance of the new developed kit was evaluated in the aspects of linearity, precision, accuracy, and specificity. Two-hundred thirty-four serum samples were enrolled to carry out the comparison test. The new PCT quantitative detecting kit exhibited a higher sensitivity (0.08 ng/mL). The inter-assay coefficient of variation (CV) and the intra-assay CV were 5.4%-7.7% and 5.7%-13.4%, respectively. The recovery rates ranged from 93% to 105%. Furthermore, a high correlation ( n = 234, r = 0.977, p LFIA was suitable for rapid and on-site detection for PCT, which might be a useful platform for other biomarkers in point-of-care tests.

  2. Rapid method for simulating gas spectra using reversed PCR temperature calibration models based on Hitran data

    DEFF Research Database (Denmark)

    Bak, J.

    1999-01-01

    A computer program was produced to make rapid simulations of CO gas spectra at a spectral resolution of 1 cm(-1) and at temperatures ranging from 295 to 845 K and concentrations from 5 to 400 mg/m(3). The program is based on loadings and scores from three principal component regression (PCR) temp...... a uniform slab of gas at various temperatures, concentrations, and pathlengths. The gain in speed of the calculations of the spectra is based on the fact that the PCR models include mathematical pretreatments and compress the data effectively.......A computer program was produced to make rapid simulations of CO gas spectra at a spectral resolution of 1 cm(-1) and at temperatures ranging from 295 to 845 K and concentrations from 5 to 400 mg/m(3). The program is based on loadings and scores from three principal component regression (PCR......) temperature calibration models. Three sets of 12 Hitran-simulated high-density spectra, each set spanning the entire temperature range at constant concentrations (50, 150, and 300 mg/m(3)), were used as calibration spectra in the PCR temperature models. All the spectra were convoluted with a sine...

  3. Selective laser sintering: application of a rapid prototyping method in craniomaxillofacial reconstructive surgery.

    Science.gov (United States)

    Aung, S C; Tan, B K; Foo, C L; Lee, S T

    1999-09-01

    Advances in technology have benefited the medical world in many ways and a new generation of computed tomography (CT) scanners and three-dimensional (3-D) model making rapid prototyping systems (RPS) have taken craniofacial surgical planning and management to new heights. With the development of new rapid prototyping systems and the improvements in CT scan technology, such as the helical scanner, biomedical modelling has improved considerably and accurate 3-D models can now be fabricated to allow surgeons to visualise and physically handle a 3-D model on which simulation surgery can be performed. The principle behind this technology is to first acquire digital data (CT scan data) which is then imported to the RPS to fabricate fine layers or cuts of the model which are gradually built up to form the 3-D models. Either liquid resin or nylon powder or special paper may be used to make these models using the various RPS available today. Selective laser sintering (SLS), which employs a CO2 laser beam to solidify special nylon powder and build up the model in layers is described in this case report, where a 23-year old Chinese female with panfacial fracture and a skull defect benefited from SLS biomodelling in the preoperative workup.

  4. Rapid determination of lipid peroxidation using a novel pyridoxamine-participating ferrous oxidation-sulfosalicylic acid spectrophotometric method.

    Science.gov (United States)

    Chen, Jingnan; Cai, Danqian; Zhang, Yu

    2016-11-15

    A novel method is developed to rapidly analyze lipid peroxidation in edible oils and fatty foods at room temperature, which is called the pyridoxamine-participating ferrous oxidation-sulfosalicylic acid (PFOS) method. The PFOS method evaluates the lipid peroxide value colorimetrically via detecting the pyridoxamine-mediated pigment produced by 5-sulfosalicylic acid and Fe(3+) at 500nm, while the latter is converted from Fe(2+) in the presence of lipid peroxides. The optimized formulation was ethanol (70%, v/v), Fe(2+) (4mmol/L), 5-sulfosalicylic acid (40mmol/L) and pyridoxamine (18mmol/L). The limit of quantitation is 0.087mmol Fe(3+)/L with acceptable reproducibility. In addition, current method has a significant linear correlation with both conventional thiobarbituric acid (R(2)=0.9999) and ferric thiocyanate assays (R(2)=0.9675). This method offers a rapid technique for evaluating lipid peroxidation without heating and sophisticated instrumental procedures. Besides, current method provides a new option to evaluate the lipid peroxidation state and improve the reproducibility of ferrous-oxidation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. A Rapid Identification Method for Calamine Using Near-Infrared Spectroscopy Based on Multi-Reference Correlation Coefficient Method and Back Propagation Artificial Neural Network.

    Science.gov (United States)

    Sun, Yangbo; Chen, Long; Huang, Bisheng; Chen, Keli

    2017-07-01

    As a mineral, the traditional Chinese medicine calamine has a similar shape to many other minerals. Investigations of commercially available calamine samples have shown that there are many fake and inferior calamine goods sold on the market. The conventional identification method for calamine is complicated, therefore as a result of the large scale of calamine samples, a rapid identification method is needed. To establish a qualitative model using near-infrared (NIR) spectroscopy for rapid identification of various calamine samples, large quantities of calamine samples including crude products, counterfeits and processed products were collected and correctly identified using the physicochemical and powder X-ray diffraction method. The NIR spectroscopy method was used to analyze these samples by combining the multi-reference correlation coefficient (MRCC) method and the error back propagation artificial neural network algorithm (BP-ANN), so as to realize the qualitative identification of calamine samples. The accuracy rate of the model based on NIR and MRCC methods was 85%; in addition, the model, which took comprehensive multiple factors into consideration, can be used to identify crude calamine products, its counterfeits and processed products. Furthermore, by in-putting the correlation coefficients of multiple references as the spectral feature data of samples into BP-ANN, a BP-ANN model of qualitative identification was established, of which the accuracy rate was increased to 95%. The MRCC method can be used as a NIR-based method in the process of BP-ANN modeling.

  6. Direct PCR - A rapid method for multiplexed detection of different serotypes of Salmonella in enriched pork meat samples.

    Science.gov (United States)

    Chin, Wai Hoe; Sun, Yi; Høgberg, Jonas; Quyen, Than Linh; Engelsmann, Pia; Wolff, Anders; Bang, Dang Duong

    2017-04-01

    Salmonellosis, an infectious disease caused by Salmonella spp., is one of the most common foodborne diseases. Isolation and identification of Salmonella by conventional bacterial culture method is time consuming. In response to the demand for rapid on line or at site detection of pathogens, in this study, we developed a multiplex Direct PCR method for rapid detection of different Salmonella serotypes directly from pork meat samples without any DNA purification steps. An inhibitor-resistant Phusion Pfu DNA polymerase was used to overcome PCR inhibition. Four pairs of primers including a pair of newly designed primers targeting Salmonella spp. at subtype level were incorporated in the multiplex Direct PCR. To maximize the efficiency of the Direct PCR, the ratio between sample and dilution buffer was optimized. The sensitivity and specificity of the multiplex Direct PCR were tested using naturally contaminated pork meat samples for detecting and subtyping of Salmonella spp. Conventional bacterial culture methods were used as reference to evaluate the performance of the multiplex Direct PCR. Relative accuracy, sensitivity and specificity of 98.8%; 97.6% and 100%, respectively, were achieved by the method. Application of the multiplex Direct PCR to detect Salmonella in pork meat at slaughter reduces the time of detection from 5 to 6 days by conventional bacterial culture and serotyping methods to 14 h (including 12 h enrichment time). Furthermore, the method poses a possibility of miniaturization and integration into a point-of-need Lab-on-a-chip system for rapid online pathogen detection. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Methods of rapid diagnosis for the etiology of meningitis in adults

    Science.gov (United States)

    Bahr, Nathan C; Boulware, David R

    2014-01-01

    Infectious meningitis may be due to bacterial, mycobacterial, fungal or viral agents. Diagnosis of meningitis must take into account numerous items of patient history and symptomatology along with regional epidemiology and basic cerebrospinal fluid testing (protein, etc.) to allow the clinician to stratify the likelihood of etiology possibilities and rationally select additional diagnostic tests. Culture is the mainstay for diagnosis in many cases, but technology is evolving to provide more rapid, reliable diagnosis. The cryptococcal antigen lateral flow assay (Immuno-Mycologics) has revolutionized diagnosis of cryptococcosis and automated nucleic acid amplification assays hold promise for improving diagnosis of bacterial and mycobacterial meningitis. This review will focus on a holistic approach to diagnosis of meningitis as well as recent technological advances. PMID:25402579

  8. Rapid Bioassessment Methods for Assessing Stream Macroinvertebrate Community on the Savannah River Site

    Energy Technology Data Exchange (ETDEWEB)

    Specht, W.L.

    1999-11-22

    Macroinvertebrate sampling was performed at 16 locations in the Savannah River Site (SRS) streams using Hester-Dendy multiplate samplers and EPA Rapid Bioassessment Protocols (RBP). Some of the sampling locations were unimpacted, while other locations had been subject to various forms of perturbation by SRS activities. In general, the data from the Hester-Dendy multiplate samplers were more sensitive at detecting impacts than were the RBP data. We developed a Biotic Index for the Hester-Dendy data which incorporated eight community structure, function, and balance parameters. when tested using a data set that was unrelated to the data set that was used in developing the Biotic Index, the index was very successful at detecting impact.

  9. HIGH YIELD AND RAPID SYNTHESES METHODS FOR PRODUCING METALLO-ORGANIC SALTS

    DEFF Research Database (Denmark)

    2005-01-01

    A new method for preparing salts of metal cations and organic acids, especially divalent salts of alkaline earth metal ions from group II of the periodic system and carboxylic acids. The method comprising the use of a high temperature (about 90° or more) and, optionally. high pressure, in order...... to obtain a higher yield, purity and faster reaction speed than obtained with known synthesis methods. In particular, the present invention relates to the production of strontium salts of carboxylic acids. Novel strontium salts are also provided by the present method....

  10. A Simple and Rapid Method for DNA Isolation from Xylophagous Insects

    Directory of Open Access Journals (Sweden)

    Horacio Cano-Camacho

    2010-12-01

    Full Text Available Published methods to isolate DNA from insects are not always effective in xylophagous insects because they have high concentrations of phenolics and other secondary plant compounds in their digestive tracts. A simple, reliable and labor-effective cetyltrimethylammonium bromide-polyvinylpyrrolidone (CTAB-PVP method for isolation of high quality DNA from xylophagous insects is described. This method was successfully applied to PCR and restriction analysis, indicating removal of common inhibitors. DNA isolated by the CTAB-PVP method could be used in most molecular analyses.

  11. Household salt iodine content estimation with the use of rapid test kits and iodometric titration methods.

    Science.gov (United States)

    Nepal, Ashwini Kumar; Raj Shakya, Prem; Gelal, Basanta; Lamsal, Madhab; Brodie, David A; Baral, Nirmal

    2013-05-01

    Universal salt iodization remains the best strategy for controlling iodine deficiency disorders in Nepal. This study was designed to study the salt types and the household salt iodine content of school aged children in the hilly and the plain districts of eastern Nepal. This cross-sectional study was carried out on school children of seven randomly chosen schools from four districts, namely, Sunsari, Dhankuta, Sankhuwasabha and Tehrathum of eastern Nepal. The school children were requested to bring two teaspoonfuls (approx. 12-15 g) of the salt which was consumed in their households, in a tightly sealed plastic pouch. The salt types were categorized, and the salt iodine content was estimated by using rapid test kits and iodometric titrations. The association of the salt iodine content of the different districts were tested by using the Chi-square test. The sensitivity, specificity, positive predictive values, and negative predictive values of the rapid test kits were compared with the iodometric titrations. Our study showed that mean±SD values of the salt iodine content in the four districts, namely, Sunsari, Dhankuta, Sankhuwasabha and Tehrathum were 34.2±17.9, 33.2±14.5, 27.4±15.1 and 48.4±15.6 parts per million (ppm). There were 270 (38.2%) households which consumed crystal salt and 437(61.8%) of the households consumed packet salts. Our study recommends a regular monitoring of the salt iodization programs in these regions. More families should be made aware of the need to ensure that each individual consumes iodized salt.

  12. Rapid assessment of insect fauna based on local knowledge: comparing ecological and ethnobiological methods.

    Science.gov (United States)

    Lima, Daniele Cristina de Oliveira; Ramos, Marcelo Alves; da Silva, Henrique Costa Hermenegildo; Alves, Angelo Giuseppe Chaves

    2016-03-01

    The rapid assessment of biodiversity making use of surveys of local knowledge has been successful for different biological taxa. However, there are no reports on the testing of such tools for sampling insect fauna. The present study aimed to evaluate the efficiency of different ethnobiological techniques for rapid sampling of insect fauna. Field research for the conventional survey of insect fauna was conducted on a private farm (9 ° 43'38.95 "S, 37 ° 45'11.97" W) , where there was intensive cultivation of okra (Abelmoschus esculentus L. (Moench)). The survey of local entomological knowledge was conducted among all the producers of okra living in the rural villages Pereira, Santa Luzia, and Nassau de Souza, within the Jacaré Curituba irrigated settlement scheme. The combined use of the techniques "free list" and projective interviews was analyzed, using two types of visual stimuli: stock photos and an entomological box. During the conventional survey of insect fauna, the species Bemisia tabaci biotype B, Aphis gossypii, Phenacoccus sp., Icerya purchasi and Lagria villosa were the primary pests found in the okra crop. Regarding the survey of insect pests, the results were convergent  in both techniques (conventional sampling and free list). Comparing the interview with visual stimuli (pictures) and specimen witnesses (entomological box) revealed that the latter was more effective. Techniques based on the recording and analysis of local knowledge about insects are effective for quick sampling of pest insects, but ineffective in sampling predator insects. The utilization of collected insects, infested branches, or photos of the symptoms of damage caused by pests in projective interviews is recommended.

  13. A simple and rapid spectrophotometric method for determination of thiophanate-methyl in commercial formulations and its residues in foodstuffs.

    Science.gov (United States)

    Verma, Balbir C; Sood, Sumita; Chauhan, Chetan; Sharma, Devender K

    2004-01-01

    A new, simple, rapid, and sensitive spectrophotometric method for the determination of thiophanate-methyl, based on its reaction with cobalt(II) in the presence of triethylamine, has been developed. The yellowish green color that develops instantaneously on mixing the fungicide with the reagents in dimethylformamide is stable for at least 2 h and has maximum absorbance at 360 nm. The method has been successfully applied to the determination of thiophanate-methyl in its commercial formulations and residues on grains and apples. A photometric titration procedure for formulation analysis of the fungicide has also been developed.

  14. Development of a rapid diagnostic method for identification of Staphylococcus aureus and antimicrobial resistance in positive blood culture bottles using a PCR-DNA-chromatography method.

    Science.gov (United States)

    Ohshiro, Takeya; Miyagi, Chihiro; Tamaki, Yoshikazu; Mizuno, Takuya; Ezaki, Takayuki

    2016-06-01

    Blood culturing and the rapid reporting of results are essential for infectious disease clinics to obtain bacterial information that can affect patient prognosis. When gram-positive coccoid cells are observed in blood culture bottles, it is important to determine whether the strain is Staphylococcus aureus and whether the strain has resistance genes, such as mecA and blaZ, for proper antibiotic selection. Previous work led to the development of a PCR method that is useful for rapid identification of bacterial species and antimicrobial susceptibility. However, that method has not yet been adopted in community hospitals due to the high cost and methodological complexity. We report here the development of a quick PCR and DNA-chromatography test, based on single-tag hybridization chromatography, that permits detection of S. aureus and the mecA and blaZ genes; results can be obtained within 1 h for positive blood culture bottles. We evaluated this method using 42 clinical isolates. Detection of S. aureus and the resistance genes by the PCR-DNA-chromatography method was compared with that obtained via the conventional identification method and actual antimicrobial susceptibility testing. Our method had a sensitivity of 97.0% and a specificity of 100% for the identification of the bacterial species. For the detection of the mecA gene of S. aureus, the sensitivity was 100% and the specificity was 95.2%. For the detection of the blaZ gene of S. aureus, the sensitivity was 100% and the specificity was 88.9%. The speed and simplicity of this PCR-DNA-chromatography method suggest that our method will facilitate rapid diagnoses. Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  15. Comparison of the efficacy of conventional slow freezing and rapid cryopreservation methods for bovine embryos

    NARCIS (Netherlands)

    Wagtendonk-de Leeuw, van A.M.; Daas, den J.H.; Kruip, T.A.; Rail, W.F.

    1995-01-01

    Day 7 bovine morulae and early blastocysts were randomly assigned to one of four cryopreservation methods: (i) a modified conventional controlled slow freezing and stepwise dilution after thawing; and three methods which enable direct transfer of the embryo into the recipient upon thawing: (ii)

  16. Colloid Titration--A Rapid Method for the Determination of Charged Colloid.

    Science.gov (United States)

    Ueno, Keihei; Kina, Ken'yu

    1985-01-01

    "Colloid titration" is a volumetric method for determining charged polyelectrolytes in aqueous solutions. The principle of colloid titration, reagents used in the procedure, methods of endpoint detection, preparation of reagent solutions, general procedure used, results obtained, and pH profile of colloid titration are considered. (JN)

  17. A simple and rapid cultural method for detection of Enterobacter sakazakii in environmental samples

    NARCIS (Netherlands)

    Guillaume-Gentil, O.; Sonnard, V.; Kandhai, M.C.; Marugg, J.; Joosten, H.

    2005-01-01

    A method was developed to detect and identify Enterobacter sakazakii in environmental samples. The method is based on selective enrichment at 45 ± 0.5°C in lauryl sulfate tryptose broth supplemented with 0.5 M NaCl and 10 mg/liter vancomycin (mLST) for 22 to 24 h followed by streaking on tryptone

  18. Rapid, sensitive and cost effective method for isolation of viral DNA from feacal samples of dogs

    Directory of Open Access Journals (Sweden)

    Savi.

    2010-06-01

    Full Text Available A simple method for viral DNA extraction using chelex resin was developed. The method used was eco-friendly and cost effective compared to other methods such as phenol chloroform method which use health hazardous organic reagents. Further, a polymerase chain reaction (PCR based detection of canine parvovirus (CPV using primers from conserved region of VP2 gene was developed. To increase the sensitivity and specificity of reaction, nested PCR was designed. PCR reaction was optimized to amplify 747bp product of VP2 gene. The assay can be completed in few hours and doesn’t need hazardous chemicals. Thus, the sample preparation using chelating resin along with nested PCR seems to be a sensitive, specific and practical method for the detection of CPV in diarrhoeal feacal samples. [Vet. World 2010; 3(3.000: 105-106

  19. A simple and rapid one-time method to evaluate the non-acidic gas content from bioprocesses.

    Science.gov (United States)

    Bassard, David; André, Laura; Dotal, Nicolas; Valentin, Ludovic; Nonus, Maurice; Pauss, André; Ribeiro, Thierry

    2014-02-01

    This paper presents a rapid less than 2 min and low-cost method involving the use of alkali solution to capture the acidic gasses from a biogas, thereby providing an estimate of the percentage of non-acidic gasses. Such a method was mentioned in the literature but never fully described or optimized. After sampling an aliquot of gas from bioprocess, gas was injected in a sealed flask with a 3 M NaOH solution, and after equilibrium was obtained, the non-acidic gas volume was measured. The method was first calibrated with certified gasses with an accuracy observed between 98 and 105%. Regarding the validation step, certified standard gas mixtures and nine biogas-laboratory batch reactors were used, the overall accuracy reported was 103 + 3%. This rapid and low-cost method may either be used in laboratory conditions as a quick and low cost alternative to standard analysis equipment or in addition as a routine field control method used on full-scale plants.

  20. Investigation of a soil stratification upper section by rapid methods of wave analysis

    Directory of Open Access Journals (Sweden)

    Antipov Vadim Valer’evich

    2016-12-01

    Full Text Available This article presents the comparative results of an investigation of the low-velocity zone of the area near the housing of the PNRPU engineering faculty by active and passive methods of multichannel analysis of surface waves (MASW. MASW is a modern nondestructive method researching the soil massif. It lets to obtain profiles of transverse velocities and to find initial elastic modulus and initial shear modulus by using these profiles. The purpose of the investigation is a study of low-velocity zone by active and passive methods of MASW. The passive method of MASW has not been used previously on the territory of Russian Federation. In the passive method, geophones have been placed by “circle,” “square” and “triangle” layouts. Also, the special case of the passive method has been applied - passive roadside MASW. The telemetry system TELSS-3 has been used to register surface Rayleigh waves. The parameters of surveying system have been selected using the optimal recommended parameters and the reconnaissance surveys. Obtained seismograms have been processed using ParkSeis software. In the stage of dispersion analysis of MASW, the dispersion images have been obtained with clearly visible fundamental modes. The obtained dispersion curves have allowed making 2D profiles of shear-wave velocities for an active method and 1D profiles for passive methods of MASW. On the basis of these profiles the models of investigated soil stratification have been obtained. The presented results allow evaluating the effectiveness of the passive method of MASW as a tool for geotechnical investigations in cramped conditions, where deployment of surveying system for the active method is impossible.

  1. Corrosion inhibitor storage and release property of TiO{sub 2} nanotube powder synthesized by rapid breakdown anodization method

    Energy Technology Data Exchange (ETDEWEB)

    Arunchandran, C.; Ramya, S.; George, R.P. [Corrosion Science and Technology Group, Indira Gandhi Centre for Atomic Research, Kalpakkam 603102 (India); Kamachi Mudali, U., E-mail: kamachi@igcar.gov.in [Corrosion Science and Technology Group, Indira Gandhi Centre for Atomic Research, Kalpakkam 603102 (India)

    2013-02-15

    Graphical abstract: Display Omitted Highlights: ► TiO{sub 2} nanotube powders were synthesized by rapid breakdown anodization method. ► Benzotriazole was loaded into the TiO{sub 2} nanotube powders. ► Low pH induced release of benzotriazole from TiO{sub 2} nanotube powders was proved. -- Abstract: Titanium dioxide (TiO{sub 2}) is one of the most studied substances in material science due to its versatile properties and diverse applications. In this study titanium dioxide nanotube powder were synthesized by rapid breakdown anodization (RBA) method. The synthesis involved potentiostatic anodization of titanium foil in 0.1 M HClO{sub 4} electrolyte under an applied voltage of 20 V and rapid stirring. The morphology and the phase of titanium dioxide nanotube powder were studied using field emission scanning electron microscopy, laser Raman spectroscopy and high resolution transmission electron microscopy. Benzotriazole was chosen as model inhibitor to evaluate TiO{sub 2} nanotube powder's corrosion inhibitor loading and releasing properties. The storage and release properties of TiO{sub 2} nanotube powder were studied using UV–visible spectroscopy and thermogravimetric analysis.

  2. Improved method for rapid and accurate isolation and identification of Streptococcus mutans and Streptococcus sobrinus from human plaque samples.

    Science.gov (United States)

    Villhauer, Alissa L; Lynch, David J; Drake, David R

    2017-08-01

    Mutans streptococci (MS), specifically Streptococcus mutans (SM) and Streptococcus sobrinus (SS), are bacterial species frequently targeted for investigation due to their role in the etiology of dental caries. Differentiation of S. mutans and S. sobrinus is an essential part of exploring the role of these organisms in disease progression and the impact of the presence of either/both on a subject's caries experience. Of vital importance to the study of these organisms is an identification protocol that allows us to distinguish between the two species in an easy, accurate, and timely manner. While conducting a 5-year birth cohort study in a Northern Plains American Indian tribe, the need for a more rapid procedure for isolating and identifying high volumes of MS was recognized. We report here on the development of an accurate and rapid method for MS identification. Accuracy, ease of use, and material and time requirements for morphological differentiation on selective agar, biochemical tests, and various combinations of PCR primers were compared. The final protocol included preliminary identification based on colony morphology followed by PCR confirmation of species identification using primers targeting regions of the glucosyltransferase (gtf) genes of SM and SS. This method of isolation and identification was found to be highly accurate, more rapid than the previous methodology used, and easily learned. It resulted in more efficient use of both time and material resources. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Using multimodal imaging techniques to monitor limb ischemia: a rapid noninvasive method for assessing extremity wounds

    Science.gov (United States)

    Luthra, Rajiv; Caruso, Joseph D.; Radowsky, Jason S.; Rodriguez, Maricela; Forsberg, Jonathan; Elster, Eric A.; Crane, Nicole J.

    2013-03-01

    Over 70% of military casualties resulting from the current conflicts sustain major extremity injuries. Of these the majority are caused by blasts from improvised explosive devices. The resulting injuries include traumatic amputations, open fractures, crush injuries, and acute vascular disruption. Critical tissue ischemia—the point at which ischemic tissues lose the capacity to recover—is therefore a major concern, as lack of blood flow to tissues rapidly leads to tissue deoxygenation and necrosis. If left undetected or unaddressed, a potentially salvageable limb may require more extensive debridement or, more commonly, amputation. Predicting wound outcome during the initial management of blast wounds remains a significant challenge, as wounds continue to "evolve" during the debridement process and our ability to assess wound viability remains subjectively based. Better means of identifying critical ischemia are needed. We developed a swine limb ischemia model in which two imaging modalities were combined to produce an objective and quantitative assessment of wound perfusion and tissue viability. By using 3 Charge-Coupled Device (3CCD) and Infrared (IR) cameras, both surface tissue oxygenation as well as overall limb perfusion could be depicted. We observed a change in mean 3CCD and IR values at peak ischemia and during reperfusion correlate well with clinically observed indicators for limb function and vitality. After correcting for baseline mean R-B values, the 3CCD values correlate with surface tissue oxygenation and the IR values with changes in perfusion. This study aims to not only increase fundamental understanding of the processes involved with limb ischemia and reperfusion, but also to develop tools to monitor overall limb perfusion and tissue oxygenation in a clinical setting. A rapid and objective diagnostic for extent of ischemic damage and overall limb viability could provide surgeons with a more accurate indication of tissue viability. This may

  4. Adaptive Channel-Tracking Method and Equalization for MC-CDMA Systems over Rapidly Fading Channel under Colored Noise

    Directory of Open Access Journals (Sweden)

    Chen Bor-Sen

    2010-01-01

    Full Text Available A recursive maximum-likelihood (RML algorithm for channel estimation under rapidly fading channel and colored noise in a multicarrier code-division multiple-access (MC-CDMA system is proposed in this paper. A moving-average model with exogenous input (MAX is given to describe the transmission channel and colored noise. Based on the pseudoregression method, the proposed RML algorithm can simultaneously estimate the parameters of channel and colored noise. Following the estimation results, these parameters can be used to enhance the minimum mean-square error (MMSE equalizer. Considering high-speed mobile stations, a one-step linear trend predictor is added to improve symbol detection. Simulation results indicate that the proposed RML estimator can track the channel more precisely than the conventional estimator. Meanwhile, the performance of the proposed enhanced MMSE equalizer is robust to the rapidly Rayleigh fading channel under colored noise in the MC-CDMA systems.

  5. Adaptive Channel-Tracking Method and Equalization for MC-CDMA Systems over Rapidly Fading Channel under Colored Noise

    Science.gov (United States)

    Yang, Chang-Yi; Chen, Bor-Sen

    2010-12-01

    A recursive maximum-likelihood (RML) algorithm for channel estimation under rapidly fading channel and colored noise in a multicarrier code-division multiple-access (MC-CDMA) system is proposed in this paper. A moving-average model with exogenous input (MAX) is given to describe the transmission channel and colored noise. Based on the pseudoregression method, the proposed RML algorithm can simultaneously estimate the parameters of channel and colored noise. Following the estimation results, these parameters can be used to enhance the minimum mean-square error (MMSE) equalizer. Considering high-speed mobile stations, a one-step linear trend predictor is added to improve symbol detection. Simulation results indicate that the proposed RML estimator can track the channel more precisely than the conventional estimator. Meanwhile, the performance of the proposed enhanced MMSE equalizer is robust to the rapidly Rayleigh fading channel under colored noise in the MC-CDMA systems.

  6. A rapid method for the determination of microbial susceptibility using the firefly luciferase assay for adenosine triphosphate (ATP)

    Science.gov (United States)

    Vellend, H.; Tuttle, S. A.; Barza, M.; Weinstein, L.; Picciolo, G. L.; Chappelle, E. W.

    1975-01-01

    Luciferase assay for adenosine triphosphate (ATP) was optimized for pure bacteria in broth in order to evaluate if changes in bacterial ATP content could be used as a rapid measure of antibiotic effect on microorganisms. Broth cultures of log phase bacteria were incubated at 310 K (37 C) for 2.5 hours at antimicrobial concentrations which resulted in the best discrimination between sensitive and resistant strains. Eighty-seven strains of 11 bacterial species were studied for their susceptibility to 12 commonly used antimicrobial agents: ampicillin, Penicillin G, nafcillin, carbenicillin, cephalothin, tetracycline, erythromycin, clindamycin, gentamicin, nitrofurantoin, colistin, and chloramplenicol. The major advantage of the ATP system over existing methods of rapid microbial susceptibility testing is that the assay can be made specific for bacterial ATP.

  7. Development of a Rapid Cell-free Method for Cytotoxicity Assessment of Vapor Phase of Cigarette Smoke

    Directory of Open Access Journals (Sweden)

    Cahours X

    2014-12-01

    Full Text Available Currently, several in vitro tests are widely used to measure toxicological properties of mainstream smoke (Neutral Red Uptake Assay, Micronucleus assay, Ames Test. These tests are necessary to assess cytotoxicity, genotoxicity, and mutagenicity, but are time consuming. This is essentially due to the preparation and the handling of cells. It is difficult to use these in vitro tests as screening method for product testing and development. For a better assessment of the cytotoxicity of the vapor phase, a rapid cell-free method has been developed. This paper describes a capillary electrophoresis cell-free method, based on the depletion of an anti-oxidant L-gamma-glutamyl-L-cysteinylglycine (GSH, applied to an aliquot of vapor phase phosphate buffered saline (PBS-trapped cigarette smoke (as recommended for in vitro testing. The correlation between this method and the survival/viability test (Neutral Red cytotoxicity is excellent (coefficient of correlation (r = 0.99.

  8. [Computer-aided method and rapid prototyping for the personalized fabrication of a silicone bandage digital prosthesis].

    Science.gov (United States)

    Ventura Ferreira, Nuno; Leal, Nuno; Correia Sá, Inês; Reis, Ana; Marques, Marisa

    2014-01-01

    The fabrication of digital prostheses has acquired growing importance not only for the possibility for the patient to overcome psychosocial trauma but also to promote grip functionality. An application method of three dimensional-computer-aided design technologies for the production of passive prostheses is presented by means of a fifth finger amputee clinical case following bilateral hand replantation.Three-dimensional-computerized tomography was used for the collection of anthropometric images of the hands. Computer-aided design techniques were used to develop the digital file-based prosthesis from the reconstruction images by inversion and superimposing the contra-lateral finger images. The rapid prototyping manufacturing method was used for the production of a silicone bandage prosthesis prototype. This approach replaces the traditional manual method by a virtual method that is basis for the optimization of a high speed, accurate and innovative process.

  9. A Rapid and Simple LC-MS Method Using Collagen Marker Peptides for Identification of the Animal Source of Leather.

    Science.gov (United States)

    Kumazawa, Yuki; Taga, Yuki; Iwai, Kenji; Koyama, Yoh-Ichi

    2016-08-03

    Identification of the animal source of leather is difficult using traditional methods, including microscopic observation and PCR. In the present study, a LC-MS method was developed for detecting interspecies differences in the amino acid sequence of type I collagen, which is a major component of leather, among six animals (cattle, horse, pig, sheep, goat, and deer). After a dechroming procedure and trypsin digestion, six tryptic peptides of type I collagen were monitored by LC-MS in multiple reaction monitoring mode for the animal source identification using the patterns of the presence or absence of the marker peptides. We analyzed commercial leathers from various production areas using this method, and found some leathers in which the commercial label disagreed with the identified animal source. Our method enabled rapid and simple leather certification and could be applied to other animals whether or not their collagen sequences are available in public databases.

  10. Rapid descriptive sensory methods – Comparison of Free Multiple Sorting, Partial Napping, Napping, Flash Profiling and conventional profiling

    DEFF Research Database (Denmark)

    Dehlholm, Christian; Brockhoff, Per B.; Meinert, Lene

    2012-01-01

    Two new rapid descriptive sensory evaluation methods are introduced to the field of food sensory evaluation. The first method, free multiple sorting, allows subjects to perform ad libitum free sortings, until they feel that no more relevant dissimilarities among products remain. The second method...... is a modal restriction of Napping to specific sensory modalities, directing sensation and still allowing a holistic approach to products. The new methods are compared to Flash Profiling, Napping and conventional descriptive sensory profiling. Evaluations are performed by several panels of expert assessors...... originating from two distinct research environments. Evaluations are performed on the same nine pâté products and within the same period of time. Results are analysed configurationally (graphically) as well as with RV coefficients, semantically and practically. Parametric bootstrapped confidence ellipses...

  11. Single-Injection HPLC Method for Rapid Analysis of a Combination Drug Delivery System

    National Research Council Canada - National Science Library

    Tucker, Robert M; Parcher, Benjamin W; Jones, Ella F; Desai, Tejal A

    2012-01-01

    .... We present a single-step method for quantifying three model therapeutics released from a model hydrogel scaffold using high-performance liquid chromatography (HPLC). Poly(ethylene glycol) dimethacrylate (PEGDMA...

  12. Flash μ-fluidics: a rapid prototyping method for fabricating microfluidic devices

    KAUST Repository

    Buttner, Ulrich

    2016-08-01

    Microfluidics has advanced in terms of design and structures; however, fabrication methods are time-consuming or expensive relative to facility costs and equipment needed. This work demonstrates a fast and economically viable 2D/3D maskless digital light-projection method based on a stereolithography process. Unlike other fabrication methods, one exposure step is used to form the whole device. Flash microfluidics is achieved by incorporating bonding and channel fabrication of complex structures in just 2.5 s to 4 s and by fabricating channel heights between 25 μm and 150 μm with photopolymer resin. The features of this fabrication technique, such as time and cost saving and easy fabrication, are used to build devices that are mostly needed in microfluidic/lab-on-chip systems. Due to the fast production method and low initial setup costs, the process could be used for point of care applications. © 2016 The Royal Society of Chemistry.

  13. Sensory and rapid instrumental methods as a combined tool for quality control of cooked ham

    Directory of Open Access Journals (Sweden)

    Sara Barbieri

    2016-11-01

    This study can be considered a first application of this combined approach that could represent a suitable and fast method to verify if the meat product purchased by consumer match its description in terms of compliance with the claimed quality.

  14. Rapid Estimation of Aircraft Performance Models using Differential Vortex Panel Method and Extended Kalman Filter Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Estimation of aerodynamic models for the control of damaged aircraft using an innovative differential vortex lattice method tightly coupled with an extended Kalman...

  15. Rapid Estimation of Aircraft Performance Models using Differential Vortex Panel Method and Extended Kalman Filter Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The problem of estimating the aerodynamic models for flight control of damaged aircraft using an innovative differential vortex lattice method tightly coupled with...

  16. Development of a rapid method to isolate polyhydroxyalkanoates from bacteria for screening studies.

    Science.gov (United States)

    Vizcaino-Caston, Isaac; Kelly, Catherine A; Fitzgerald, Annabel V L; Leeke, Gary A; Jenkins, Mike; Overton, Tim W

    2016-01-01

    We describe a novel method of Polyhydroxyalkanoate (PHA) extraction using dimethyl sulphoxide (DMSO) for use in screening studies. Compared to conventional chloroform extraction, the DMSO method was shown to release comparable quantities of PHA from Cupriavidus necator cells, with comparable properties as determined using Fourier transform infrared spectroscopy and differential scanning calorimetry. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  17. A convenient method for rapid determination of proteins in rices and rice products

    OpenAIRE

    光永, 俊郎; 安藤, ひとみ

    1990-01-01

    [Author abstract]In the biuret determination of protein in rices and rice products, starch and lipids extractable with an alkaline solution such as biuret reagent were found to interfere with the biuret method. Comparative evaluation of their effects revealed that starch, a major component of rice, has the most significant effect on the biuret method. In the presence of starch, the correlation between Kjeldahl protein and the optical density of biuret was poor. It was found that several perha...

  18. A rapid chemical method for lysing Arabidopsis cells for protein analysis

    OpenAIRE

    Takano Tetsuo; Liu Shenkui; Tsugama Daisuke

    2011-01-01

    Abstract Background Protein extraction is a frequent procedure in biological research. For preparation of plant cell extracts, plant materials usually have to be ground and homogenized to physically break the robust cell wall, but this step is laborious and time-consuming when a large number of samples are handled at once. Results We developed a chemical method for lysing Arabidopsis cells without grinding. In this method, plants are boiled for just 10 minutes in a solution containing a Ca2+ ...

  19. Rapid quantitative analysis of lipids using a colorimetric method in a microplate format.

    Science.gov (United States)

    Cheng, Yu-Shen; Zheng, Yi; VanderGheynst, Jean S

    2011-01-01

    A colorimetric sulfo-phospho-vanillin (SPV) method was developed for high throughput analysis of total lipids. The developed method uses a reaction mixture that is maintained in a 96-well microplate throughout the entire assay. The new assay provides the following advantages over other methods of lipid measurement: (1) background absorbance can be easily corrected for each well, (2) there is less risk of handling and transferring sulfuric acid contained in reaction mixtures, (3) color develops more consistently providing more accurate measurement of absorbance, and (4) the assay can be used for quantitative measurement of lipids extracted from a wide variety of sources. Unlike other spectrophotometric approaches that use fluorescent dyes, the optimal spectra and reaction conditions for the developed assay do not vary with the sample source. The developed method was used to measure lipids in extracts from four strains of microalgae. No significant difference was found in lipid determination when lipid content was measured using the new method and compared to results obtained using a macro-gravimetric method.

  20. A Rapid Coordinate Transformation Method Applied in Industrial Robot Calibration Based on Characteristic Line Coincidence

    Directory of Open Access Journals (Sweden)

    Bailing Liu

    2016-02-01

    Full Text Available Coordinate transformation plays an indispensable role in industrial measurements, including photogrammetry, geodesy, laser 3-D measurement and robotics. The widely applied methods of coordinate transformation are generally based on solving the equations of point clouds. Despite the high accuracy, this might result in no solution due to the use of ill conditioned matrices. In this paper, a novel coordinate transformation method is proposed, not based on the equation solution but based on the geometric transformation. We construct characteristic lines to represent the coordinate systems. According to the space geometry relation, the characteristic line scan is made to coincide by a series of rotations and translations. The transformation matrix can be obtained using matrix transformation theory. Experiments are designed to compare the proposed method with other methods. The results show that the proposed method has the same high accuracy, but the operation is more convenient and flexible. A multi-sensor combined measurement system is also presented to improve the position accuracy of a robot with the calibration of the robot kinematic parameters. Experimental verification shows that the position accuracy of robot manipulator is improved by 45.8% with the proposed method and robot calibration.

  1. Ecosytem Services: A Rapid Assessment Method Tested at 35 Sites of the LTER-Europe Network

    Directory of Open Access Journals (Sweden)

    Dick Jan

    2014-08-01

    Full Text Available The identification of parameters to monitor the ecosystem services delivered at a site is fundamental to the concept’s adoption as a useful policy instrument at local, national and international scales. In this paper we (i describe the process of developing a rapid comprehensive ecosystem service assessment methodology and (ii test the applicability of the protocol at 35 long-term research (LTER sites across 14 countries in the LTER-Europe network (www.lter-europe.net including marine, urban, agricultural, forest, desert and conservation sites. An assessment of probability of occurrence with estimated confidence score using 83 ecosystem service parameters was tested. The parameters were either specific services like food production or proxies such as human activities which were considered surrogates for cultural diversity and economic activity. This initial test of the ecosystem service parameter list revealed that the parameters tested were relatively easy to score by site managers with a high level of certainty (92% scored as either occurring or not occurring at the site with certainty of over 90%. Based on this assessment, we concluded that (i this approach to operationalise the concept of ecosystem services is practical and applicable by many sectors of civil society as a first screen of the ecosystem services present at a site, (ii this study has direct relevance to land management and policy decision makers as a transparent vehicle to focus testing scenarios and target data gathering, but (iii further work beyond the scale investigated here is required to ensure global applicability.

  2. A rapid direct solvent extraction method for the extraction of 2-dodecylcyclobutanone from irradiated ground beef patties using acetonitrile.

    Science.gov (United States)

    Hijaz, Faraj; Kumar, Amit; Smith, J Scott

    2010-08-01

    The amount of irradiated beef in the U.S. market is growing, and a reliable, rapid method is needed to detect irradiated beef and quantify the irradiation dose. The official analytical method (BS EN 1785 2003) that has been adopted by the European Union is time consuming. The objective of this study was to develop a rapid method for the analysis of 2-dodecylcyclobutanone (2-DCB) in irradiated beef. A 5 g sample of commercially irradiated ground beef patty (90/10) was extracted with n-hexane using a Soxhlet apparatus or with acetonitrile via direct solvent extraction. The Soxhlet hexane extract was evaporated to dryness, and the sample was dissolved in a mixture of ethyl acetate and acetonitrile (1:1). The defatted extract was purified with a 1 g silica cartridge. Another 5 g aliquot of the same patty was mixed with 50 mL acetonitrile and either blended for 1 min with a hand blender or crushed for 10 min with a glass rod. The extraction procedure was repeated 3 times, and the acetonitrile was collected and evaporated to dryness. Eluants from both methods were concentrated under nitrogen and injected into a gas chromatography-mass spectrometry. The 2-DCB concentration in the commercial samples was 0.031 +/- 0.0026 ppm (n = 5) for the Soxhlet method and 0.031 +/- 0.0025 ppm (n = 10) for direct solvent extraction. Recovery of 2-DCB from spiked beef samples in the direct solvent extraction method was 93.2 +/- 9.0% (n = 7). This study showed that the direct solvent extraction method is simple and as efficient and reproducible as the Soxhlet method.

  3. Method Development for Rapid Analysis of Natural Radioactive Nuclides Using Sector Field Inductively Coupled Plasma Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Lim, J.M.; Ji, Y.Y.; Lee, H.; Park, J.H.; Jang, M.; Chung, K.H.; Kang, M.J.; Choi, G.S. [Korea Atomic Energy Research Institute (Korea, Republic of)

    2014-07-01

    As an attempt to reduce the social costs and apprehension arising from radioactivity in the environment, an accurate and rapid assessment of radioactivity is highly desirable. Naturally occurring radioactive materials (NORM) are widely spread throughout the environment. The concern with radioactivity from these materials has therefore been growing for the last decade. In particular, radiation exposure in the industry when handling raw materials (e.g., coal mining and combustion, oil and gas production, metal mining and smelting, mineral sands (REE, Ti, Zr), fertilizer (phosphate), and building materials) has been brought to the public's attention. To decide the proper handling options, a rapid and accurate analytical method that can be used to evaluate the radioactivity of radionuclides (e.g., {sup 238}U, {sup 235}U, {sup 232}Th, {sup 226}Ra, and {sup 40}K) should be developed and validated. Direct measuring methods such as alpha spectrometry, a liquid scintillation counter (LSC), and mass-spectrometry are usually used for the measurement of radioactivity in NORM samples, and they encounter the most significant difficulties during pretreatment (e.g., purification, speciation, and dilution/enrichment). Since the pretreatment process consequently plays an important role in the measurement uncertainty, method development and validation should be performed. Furthermore, a-spectrometry has a major disadvantage of a long counting time, while it has a prominent measurement capability at a very low activity level of {sup 238}U, {sup 235}U, {sup 232}Th, and {sup 226}Ra. Contrary to the α-spectrometry method, a measurement technique using ICP-MS allow radioactivity in many samples to be measured in a short time period with a high degree of accuracy and precision. In this study, a method was developed for a rapid analysis of natural radioactive nuclides using ICP-MS. A sample digestion process was established using LiBO{sub 2} fusion and Fe co-precipitation. A magnetic

  4. Fat and Moisture Content in Chinese Fried Bread Sticks: Assessment and Rapid Near-Infrared Spectroscopic Method Development

    Directory of Open Access Journals (Sweden)

    Zhuqing Xiao

    2013-01-01

    Full Text Available Fried bread sticks (FBS are one of the most widely consumed deep fried food products in China. Understanding the fat and moisture content in FBS will help consumers make healthy food choices as well as assist food processors to provide FBS with desirable quality. Rapid Fourier transform near-infrared methods (FT-NIR were developed for determining fat and moisture content in FBS collected from 123 different vendors in Shanghai, China. FBS samples with minimum sample preparation (either finely or coarsely ground were used for NIR analyses. Spectra of FBS were treated with different mathematic pretreatments before being used to build models between the spectral information and fat (7.71%–30.89% or moisture (17.39%–32.65% content in FBS. Finely ground samples may lead to slightly more robust PLS models, but the particle sizes of ground FBS samples did not seriously affect the predictability of the models with appropriate mathematical treatments. The fat and moisture content in FBS predicted by FT-NIR methods had very good correlation with their values determined via traditional methods (fat, R2=0.965; moisture, R2=0.983, which clearly indicated that FT-NIR methods could be used as an effective tool for rapid determination of fat and moisture content in FBS.

  5. Fast mouse PK (Fast PK): a rapid screening method to increase pharmacokinetic throughput in pre-clinical drug discovery.

    Science.gov (United States)

    Reddy, Jitendar; Madishetti, Sreedhar; Vachaspati, Prakash R

    2012-09-29

    We describe a rapid screening methodology for performing pharmacokinetic (PK) studies in mice called Fast PK. In this Fast PK method, two mice were used per compound and four blood samples were collected from each mouse. The sampling times were staggered (sparse sampling) between the two mice, thus yielding complete PK profile in singlicate across eight time points. The plasma PK parameters from Fast PK were comparable to that obtained from conventional PK methods. This method has been used to rapidly screen compounds in the early stages of drug discovery and about 600 compounds have been profiled in the last 3 years, which has resulted in reduction in the usage of mice by 800 per year in compliance with the 3R principles of animal ethics. In addition, this Fast PK method can also help in evaluating the PK parameters from the same set of animals used in safety/toxicology/efficacy studies without the need for satellite groups. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. A Flow Cytometry Method for Rapidly Assessing Mycobacterium tuberculosis Responses to Antibiotics with Different Modes of Action.

    Science.gov (United States)

    Hendon-Dunn, Charlotte Louise; Doris, Kathryn Sarah; Thomas, Stephen Richard; Allnutt, Jonathan Charles; Marriott, Alice Ann Neville; Hatch, Kim Alexandra; Watson, Robert James; Bottley, Graham; Marsh, Philip David; Taylor, Stephen Charles; Bacon, Joanna

    2016-07-01

    Current methods for assessing the drug susceptibility of Mycobacterium tuberculosis are lengthy and do not capture information about viable organisms that are not immediately culturable under standard laboratory conditions as a result of antibiotic exposure. We have developed a rapid dual-fluorescence flow cytometry method using markers for cell viability and death. We show that the fluorescent marker calcein violet with an acetoxy-methyl ester group (CV-AM) can differentiate between populations of M. tuberculosis growing at different rates, while Sytox green (SG) can differentiate between live and dead mycobacteria. M. tuberculosis was exposed to isoniazid or rifampin at different concentrations over time and either dual stained with CV-AM and SG and analyzed by flow cytometry or plated to determine the viability of the cells. Although similar trends in the loss of viability were observed when the results of flow cytometry and the plate counting methods were compared, there was a lack of correlation between these two approaches, as the flow cytometry analysis potentially captured information about cell populations that were unable to grow under standard conditions. The flow cytometry approach had an additional advantage in that it could provide insights into the mode of action of the drug: antibiotics targeting the cell wall gave a flow cytometry profile distinct from those inhibiting intracellular processes. This rapid drug susceptibility testing method could identify more effective antimycobacterials, provide information about their potential mode of action, and accelerate their progress to the clinic. Copyright © 2016 Hendon-Dunn et al.

  7. Rapid Salmonella detection using an acoustic wave device combined with the RCA isothermal DNA amplification method

    Directory of Open Access Journals (Sweden)

    Antonis Kordas

    2016-12-01

    Full Text Available Salmonella enterica serovar Typhimurium is a major foodborne pathogen that causes Salmonellosis, posing a serious threat for public health and economy; thus, the development of fast and sensitive methods is of paramount importance for food quality control and safety management. In the current work, we are presenting a new approach where an isothermal amplification method is combined with an acoustic wave device for the development of a label free assay for bacteria detection. Specifically, our method utilizes a Love wave biosensor based on a Surface Acoustic Wave (SAW device combined with the isothermal Rolling Circle Amplification (RCA method; various protocols were tested regarding the DNA amplification and detection, including off-chip amplification at two different temperatures (30 °C and room temperature followed by acoustic detection and on-chip amplification and detection at room temperature, with the current detection limit being as little as 100 Bacteria Cell Equivalents (BCE/sample. Our acoustic results showed that the acoustic ratio, i.e., the amplitude over phase change observed during DNA binding, provided the only sensitive means for product detection while the measurement of amplitude or phase alone could not discriminate positive from negative samples. The method's fast analysis time together with other inherent advantages i.e., portability, potential for multi-analysis, lower sample volumes and reduced power consumption, hold great promise for employing the developed assay in a Lab on Chip (LoC platform for the integrated analysis of Salmonella in food samples.

  8. Rapid and Efficient Method for the Detection of Microplastic in the Gastrointestinal Tract of Fishes.

    Science.gov (United States)

    Roch, Samuel; Brinker, Alexander

    2017-04-18

    The rising evidence of microplastic pollution impacts on aquatic organisms in both marine and freshwater ecosystems highlights a pressing need for adequate and comparable detection methods. Available tissue digestion protocols are time-consuming (>10 h) and/or require several procedural steps, during which materials can be lost and contaminants introduced. This novel approach comprises an accelerated digestion step using sodium hydroxide and nitric acid in combination to digest all organic material within 1 h plus an additional separation step using sodium iodide which can be used to reduce mineral residues in samples where necessary. This method yielded a microplastic recovery rate of ≥95%, and all tested polymer types were recovered with only minor changes in weight, size, and color with the exception of polyamide. The method was also shown to be effective on field samples from two benthic freshwater fish species, revealing a microplastic burden comparable to that indicated in the literature. As a consequence, the present method saves time, minimizes the loss of material and the risk of contamination, and facilitates the identification of plastic particles and fibers, thus providing an efficient method to detect and quantify microplastics in the gastrointestinal tract of fishes.

  9. Rapid Retrieval of Lung Nodule CT Images Based on Hashing and Pruning Methods

    Directory of Open Access Journals (Sweden)

    Ling Pan

    2016-01-01

    Full Text Available The similarity-based retrieval of lung nodule computed tomography (CT images is an important task in the computer-aided diagnosis of lung lesions. It can provide similar clinical cases for physicians and help them make reliable clinical diagnostic decisions. However, when handling large-scale lung images with a general-purpose computer, traditional image retrieval methods may not be efficient. In this paper, a new retrieval framework based on a hashing method for lung nodule CT images is proposed. This method can translate high-dimensional image features into a compact hash code, so the retrieval time and required memory space can be reduced greatly. Moreover, a pruning algorithm is presented to further improve the retrieval speed, and a pruning-based decision rule is presented to improve the retrieval precision. Finally, the proposed retrieval method is validated on 2,450 lung nodule CT images selected from the public Lung Image Database Consortium (LIDC database. The experimental results show that the proposed pruning algorithm effectively reduces the retrieval time of lung nodule CT images and improves the retrieval precision. In addition, the retrieval framework is evaluated by differentiating benign and malignant nodules, and the classification accuracy can reach 86.62%, outperforming other commonly used classification methods.

  10. Rapid Retrieval of Lung Nodule CT Images Based on Hashing and Pruning Methods.

    Science.gov (United States)

    Pan, Ling; Qiang, Yan; Yuan, Jie; Wu, Lidong

    2016-01-01

    The similarity-based retrieval of lung nodule computed tomography (CT) images is an important task in the computer-aided diagnosis of lung lesions. It can provide similar clinical cases for physicians and help them make reliable clinical diagnostic decisions. However, when handling large-scale lung images with a general-purpose computer, traditional image retrieval methods may not be efficient. In this paper, a new retrieval framework based on a hashing method for lung nodule CT images is proposed. This method can translate high-dimensional image features into a compact hash code, so the retrieval time and required memory space can be reduced greatly. Moreover, a pruning algorithm is presented to further improve the retrieval speed, and a pruning-based decision rule is presented to improve the retrieval precision. Finally, the proposed retrieval method is validated on 2,450 lung nodule CT images selected from the public Lung Image Database Consortium (LIDC) database. The experimental results show that the proposed pruning algorithm effectively reduces the retrieval time of lung nodule CT images and improves the retrieval precision. In addition, the retrieval framework is evaluated by differentiating benign and malignant nodules, and the classification accuracy can reach 86.62%, outperforming other commonly used classification methods.

  11. A Rapid Empirical Method for Estimating the Gross Takeoff Weight of a High Speed Civil Transport

    Science.gov (United States)

    Mack, Robert J.

    1999-01-01

    During the cruise segment of the flight mission, aircraft flying at supersonic speeds generate sonic booms that are usually maximum at the beginning of cruise. The pressure signature with the shocks causing these perceived booms can be predicted if the aircraft's geometry, Mach number, altitude, angle of attack, and cruise weight are known. Most methods for estimating aircraft weight, especially beginning-cruise weight, are empirical and based on least- square-fit equations that best represent a body of component weight data. The empirical method discussed in this report used simplified weight equations based on a study of performance and weight data from conceptual and real transport aircraft. Like other weight-estimation methods, weights were determined at several points in the mission. While these additional weights were found to be useful, it is the determination of beginning-cruise weight that is most important for the prediction of the aircraft's sonic-boom characteristics.

  12. Method for rapid, controllable growth and thickness, of epitaxial silicon films

    Science.gov (United States)

    Wang, Qi [Littleton, CO; Stradins, Paul [Golden, CO; Teplin, Charles [Boulder, CO; Branz, Howard M [Boulder, CO

    2009-10-13

    A method of producing epitaxial silicon films on a c-Si wafer substrate using hot wire chemical vapor deposition by controlling the rate of silicon deposition in a temperature range that spans the transition from a monohydride to a hydrogen free silicon surface in a vacuum, to obtain phase-pure epitaxial silicon film of increased thickness is disclosed. The method includes placing a c-Si substrate in a HWCVD reactor chamber. The method also includes supplying a gas containing silicon at a sufficient rate into the reaction chamber to interact with the substrate to deposit a layer containing silicon thereon at a predefined growth rate to obtain phase-pure epitaxial silicon film of increased thickness.

  13. Preparation and Characterization of Micronized Artemisinin via a Rapid Expansion of Supercritical Solutions (RESS Method

    Directory of Open Access Journals (Sweden)

    Xiaonan Zhang

    2012-04-01

    Full Text Available The particle sizes of pharmaceutical substances are important for their bioavailability. Bioavailability can be improved by reducing the particle size of the drug. In this study, artemisinin was micronized by the rapid expansion of supercritical solutions (RESS. The particle size of the unprocessed white needle-like artemisinin particles was 30 to 1200 µm. The optimum micronization conditions are determined as follows: extraction temperature of 62 °C, extraction pressure of 25 MPa, precipitation temperature 45 °C and nozzle diameter of 1000 μm. Under the optimum conditions, micronized artemisinin with a (mean particle size MPS of 550 nm is obtained. By analysis of variance (ANOVA, extraction temperature and pressure have significant effects on the MPS of the micronized artemisinin. The particle size of micronized artemisinin decreased with increasing extraction temperature and pressure. Moreover, the SEM, LC-MS, FTIR, DSC and XRD allowed the comparison between the crystalline initial state and the micronization particles obtained after the RESS process. The results showed that RESS process has not induced degradation of artemisinin and that processed artemisinin particles have lower crystallinity and melting point. The bulk density of artemisinin was determined before and after RESS process and the obtained results showed that it passes from an initial density of 0.554 to 0.128 g·cm−3 after the processing. The decrease in bulk density of the micronized powder can increase the liquidity of drug particles when they are applied for medicinal preparations. These results suggest micronized powder of artemisinin can be of great potential in drug delivery systems.

  14. Ball mill assisted rapid mechanochemical extraction method for natural products from plants.

    Science.gov (United States)

    Wang, Man; Bi, Wentao; Huang, Xiaohua; Chen, David Da Yong

    2016-06-03

    A ball mill assisted mechanochemical extraction method was developed to extract compounds of natural product (NP) from plant using ionic liquid (IL). A small volume ball mill, also known as PastPrep(®) Homogenizer, which is often used for high-speed lysis of biological samples and for other applications, was used to dramatically increase the speed, completeness and reproducibility of the extraction process at room temperature to preserve the chemical integrity of the extracted compounds. In this study, tanshinones were selected as target compounds to evaluate the performance of this extraction method. Factors affecting the extraction efficiency, such as the duration, IL concentration and solid/liquid ratio were systematically optimized using the response surface methodology. Under the optimized conditions, the described method was more efficient and much faster than the conventional extraction methods such as methanol based ultrasound assisted extraction (UAE) and heat reflux extraction (HRE) that consumes a lot more organic solvent. In addition, the natural products of interest were enriched by anion metathesis of ionic liquids, combining extraction and preconcentration in the same process. The extractant was analyzed by HPLC and LC-MS. The reproducibility (RSD, n=5), correlation coefficient (r(2)) of the calibration curve, and the limit of detection, were determined to be in the range of 4.7-5.2%, 0.9992-0.9995, and 20-51ng/mL, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. New rapid method for determining edgewise compressive strength of corrugated fiberboard

    Science.gov (United States)

    John W. Koning

    1986-01-01

    The objective of this study was to determine if corrugated fiberboard specimens that had been necked down with a common router would yield acceptable edgewise compressive strength values. Tests were conducted on specimens prepared using a circular saw and router, and the results were compared with those obtained on specimens prepared according to TAPPI Test Method T...

  16. Estimation of plant-available nitrogen in soils using rapid chemical and biological methods

    NARCIS (Netherlands)

    Velthof, G.L.; Oenema, O.

    2010-01-01

    The relationships between potential laboratory indices for plant-available nitrogen (N) and the plant N uptake in a pot experiment with ryegrass were assessed for 13 mineral soils and 2 peat soils. The methods included aerobic soil incubation, soil incubation in a bioreactor, hot potassium chloride

  17. Rapid deletion plasmid construction methods for protoplast and Agrobacterium based fungal transformation systems

    Science.gov (United States)

    Increasing availability of genomic data and sophistication of analytical methodology in fungi has elevated the need for functional genomics tools in these organisms. Gene deletion is a critical tool for functional analysis. The targeted deletion of genes requires both a suitable method for the trans...

  18. Profiling method for a rapid and efficient screening of transgenic plants

    African Journals Online (AJOL)

    user

    2012-06-28

    Jun 28, 2012 ... Metabolomics-based approaches are methods of choice for studying changes in fruit composition induced by environmental or genetic modulation of biochemical pathways in the fruit. Owing to enzyme redundancy and high plasticity of the metabolic network, transgenic alteration of the activity of the.

  19. Rapid method for the determination of metamizole residues in bovine muscle by LC-MS/MS.

    Science.gov (United States)

    Jedziniak, Piotr; Pietruk, Konrad; Śledzińska, Edyta; Olejnik, Małgorzata; Szprengier-Juszkiewicz, Teresa; Żmudzki, Jan

    2013-01-01

    Metamizole is a pyrazolone non-steroidal anti-inflammatory drug allowed for use in food-producing animals. According to Council Directive 96/23, residues of this drug have to be monitored because of the potential risk to consumers' health. Metamizole is hydrolysed to its marker residue 4-methylaminoantypyrine.This compound is further metabolised to three main metabolites: 4-formylaminoantipyrine, 4-aminoantipyrine and 4-acetylaminoantipyrine. The MRL of 4-methylaminoantipyrine in animal tissues is 100 µg kg(-1). Considering the above points, a method for the detection of four metamizole metabolites in bovine muscles was developed. Analytes were extracted from muscle by a mixture of acetonitrile and sodium acetate buffer. After centrifugation, the supernatant was passed through alumina cartridges, diluted with mobile phase and analysed by using LC-MS/MS. Four metamizole metabolites were separated on a C8 column in 23 min with a gradient of methanol:acetonitrile:ammonium formate solution and analysed by using positive ionisation. Validation of the method indicated a within-laboratory reproducibility in the range of 7-30% and recovery in the range of 45-95%. The method fulfils the criteria for confirmatory methods and, thanks to its labour efficiency, may also be used for screening purposes.

  20. Evaluation of a membrane filtration method for the rapid enumeration of confirmed Clostridium perfringens from water.

    Science.gov (United States)

    Watkins, J; Sartory, D P

    2015-04-01

    A modification of the UK reference and ISO 14189 TSCA medium for the enumeration of Clostridium perfringens from water coupled with a membrane filter transfer technique for testing for production of acid phosphatase was evaluated. The new tryptose cycloserine agar (TCA) medium, which lacks sodium metabisulphite but contains sodium pyruvate to improve recovery, allows the isolation and confirmation of Cl. perfringens within 18-24 h of sample processing. Data from a multilaboratory study analysed according to ISO 17994 showed that TCA was equivalent to TSCA for the enumeration of Cl. perfringens. The identification of acid phosphatase-negative isolates revealed a false-negative rate for the TCA method of 0.8%. The TCA membrane filter transfer procedure provides confirmed Cl. perfringens counts in half the time of the TSCA method and is simple to undertake. The testing of drinking water for Clostridium perfringens is a regulatory parameter in Europe and the UK. Current UK and ISO methods employ membrane filtration (MF) and TSCA medium followed by subculture and confirmation of isolates by testing for acid phosphatase. This takes 48 h. We present here the results of a multilaboratory evaluation of a MF method that features a simplified isolation medium (TCA) and a membrane transfer procedure for the acid phosphatase test resulting in confirmed results being available in 18-24 h. This development significantly reduces the time to confirmed results for Cl. perfringens from water samples. © 2014 The Society for Applied Microbiology.

  1. A method for rapid quantitative assessment of biofilms with biomolecular staining and image analysis.

    Science.gov (United States)

    Larimer, Curtis; Winder, Eric; Jeters, Robert; Prowant, Matthew; Nettleship, Ian; Addleman, Raymond Shane; Bonheyo, George T

    2016-01-01

    The accumulation of bacteria in surface-attached biofilms can be detrimental to human health, dental hygiene, and many industrial processes. Natural biofilms are soft and often transparent, and they have heterogeneous biological composition and structure over micro- and macroscales. As a result, it is challenging to quantify the spatial distribution and overall intensity of biofilms. In this work, a new method was developed to enhance the visibility and quantification of bacterial biofilms. First, broad-spectrum biomolecular staining was used to enhance the visibility of the cells, nucleic acids, and proteins that make up biofilms. Then, an image analysis algorithm was developed to objectively and quantitatively measure biofilm accumulation from digital photographs and results were compared to independent measurements of cell density. This new method was used to quantify the growth intensity of Pseudomonas putida biofilms as they grew over time. This method is simple and fast, and can quantify biofilm growth over a large area with approximately the same precision as the more laborious cell counting method. Stained and processed images facilitate assessment of spatial heterogeneity of a biofilm across a surface. This new approach to biofilm analysis could be applied in studies of natural, industrial, and environmental biofilms.

  2. A single-step method for rapid extraction of total lipids from green microalgae.

    Directory of Open Access Journals (Sweden)

    Martin Axelsson

    Full Text Available Microalgae produce a wide range of lipid compounds of potential commercial interest. Total lipid extraction performed by conventional extraction methods, relying on the chloroform-methanol solvent system are too laborious and time consuming for screening large numbers of samples. In this study, three previous extraction methods devised by Folch et al. (1957, Bligh and Dyer (1959 and Selstam and Öquist (1985 were compared and a faster single-step procedure was developed for extraction of total lipids from green microalgae. In the single-step procedure, 8 ml of a 2∶1 chloroform-methanol (v/v mixture was added to fresh or frozen microalgal paste or pulverized dry algal biomass contained in a glass centrifuge tube. The biomass was manually suspended by vigorously shaking the tube for a few seconds and 2 ml of a 0.73% NaCl water solution was added. Phase separation was facilitated by 2 min of centrifugation at 350 g and the lower phase was recovered for analysis. An uncharacterized microalgal polyculture and the green microalgae Scenedesmus dimorphus, Selenastrum minutum, and Chlorella protothecoides were subjected to the different extraction methods and various techniques of biomass homogenization. The less labour intensive single-step procedure presented here allowed simultaneous recovery of total lipid extracts from multiple samples of green microalgae with quantitative yields and fatty acid profiles comparable to those of the previous methods. While the single-step procedure is highly correlated in lipid extractability (r² = 0.985 to the previous method of Folch et al. (1957, it allowed at least five times higher sample throughput.

  3. A rapid and robust method of identifying transformed Arabidopsis thaliana seedlings following floral dip transformation

    Directory of Open Access Journals (Sweden)

    Gray John C

    2006-11-01

    Full Text Available Abstract Background The floral dip method of transformation by immersion of inflorescences in a suspension of Agrobacterium is the method of choice for Arabidopsis transformation. The presence of a marker, usually antibiotic- or herbicide-resistance, allows identification of transformed seedlings from untransformed seedlings. Seedling selection is a lengthy process which does not always lead to easily identifiable transformants. Selection for kanamycin-, phosphinothricin- and hygromycin B-resistance commonly takes 7–10 d and high seedling density and fungal contamination may result in failure to recover transformants. Results A method for identifying transformed seedlings in as little as 3.25 d has been developed. Arabidopsis T1 seeds obtained after floral dip transformation are plated on 1% agar containing MS medium and kanamycin, phosphinothricin or hygromycin B, as appropriate. After a 2-d stratification period, seeds are subjected to a regime of 4–6 h light, 48 h dark and 24 h light (3.25 d. Kanamycin-resistant and phosphinothricin-resistant seedlings are easily distinguished from non-resistant seedlings by green expanded cotyledons whereas non-resistant seedlings have pale unexpanded cotyledons. Seedlings grown on hygromycin B differ from those grown on kanamycin and phosphinothricin as both resistant and non-resistant seedlings are green. However, hygromycin B-resistant seedlings are easily identified as they have long hypocotyls (0.8–1.0 cm whereas non-resistant seedlings have short hypocotyls (0.2–0.4 cm. Conclusion The method presented here is an improvement on current selection methods as it allows quicker identification of transformed seedlings: transformed seedlings are easily discernable from non-transformants in as little as 3.25 d in comparison to the 7–10 d required for selection using current protocols.

  4. A rapid method for creating qualitative images indicative of thick oil emulsion on the ocean's surface from imaging spectrometer data

    Science.gov (United States)

    Kokaly, Raymond F.; Hoefen, Todd M.; Livo, K. Eric; Swayze, Gregg A.; Leifer, Ira; McCubbin, Ian B.; Eastwood, Michael L.; Green, Robert O.; Lundeen, Sarah R.; Sarture, Charles M.; Steele, Denis; Ryan, Thomas; Bradley, Eliza S.; Roberts, Dar A.; ,

    2010-01-01

    This report describes a method to create color-composite images indicative of thick oil:water emulsions on the surface of clear, deep ocean water by using normalized difference ratios derived from remotely sensed data collected by an imaging spectrometer. The spectral bands used in the normalized difference ratios are located in wavelength regions where the spectra of thick oil:water emulsions on the ocean's surface have a distinct shape compared to clear water and clouds. In contrast to quantitative analyses, which require rigorous conversion to reflectance, the method described is easily computed and can be applied rapidly to radiance data or data that have been atmospherically corrected or ground-calibrated to reflectance. Examples are shown of the method applied to Airborne Visible/Infrared Imaging Spectrometer data collected May 17 and May 19, 2010, over the oil spill from the Deepwater Horizon offshore oil drilling platform in the Gulf of Mexico.

  5. Notch stress intensity factors under mixed mode loadings: an overview of recent advanced methods for rapid calculation

    Directory of Open Access Journals (Sweden)

    M. Peron

    2017-10-01

    Full Text Available Recently some methods for the rapid calculation of notch stress intensity factors (NSIFs have been developed and three of them are compared in this work. First, the criteria proposed by Lazzarin et al. and Treifi et al. have been reviewed. The former is based on the calculation of the mean value of SED on two different control volume (characterized by two different radius values centred at the stress singularity point, whereas the latter takes advantage of the strain energy density averaged within two control volumes (semi-circular sector centred at the notch tip. Then, a new method based on the evaluation of the total and deviatoric SED averaged in a single control volume has been proposed. Finally, plate specimens weakened by different notch geometries have been subjected to the application of the above mentioned methods and the obtained values of the NSIFs have been compared with those derived according to Gross and Mendelson

  6. A novel stent inflation protocol improves long-term outcomes compared with rapid inflation/deflation deployment method.

    Science.gov (United States)

    Vallurupalli, Srikanth; Kasula, Srikanth; Kumar Agarwal, Shiv; Pothineni, Naga Venkata K; Abualsuod, Amjad; Hakeem, Abdul; Ahmed, Zubair; Uretsky, Barry F

    2017-08-01

    High-pressure inflation for coronary stent deployment is universally performed. However, the duration of inflation is variable and does not take into account differences in lesion compliance. We developed a standardized "pressure optimization protocol" (POP) using inflation pressure stability rather than an arbitrary inflation time or angiographic balloon appearance for stent deployment. Whether this approach improves long-term outcomes is unknown. 792 patients who underwent PCI using either rapid inflation/deflation (n = 376) or POP (n = 416) between January 2009 and March 2014 were included. Exclusion criteria included PCI for acute myocardial infarction, in-stent restenosis, chronic total occlusion, left main, and saphenous vein graft lesions. Primary endpoint was target vessel failure [TVF = combined end point of target vessel revascularization (TVR), myocardial infarction, and cardiac death]. Outcomes were analyzed in the entire cohort and in a propensity analysis. Stent implantation using POP with a median follow-up of 1317 days was associated with lower TVF compared with rapid inflation/deflation (10.1 vs. 17.8%, P inflation/deflation (10 vs. 18%, P < 0.0001). Stent deployment using POP led to reduced TVF compared to rapid I/D. These results recommend this method to improve long-term outcomes. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  7. A new rapid method for Clostridium difficile DNA extraction and detection in stool: toward point-of-care diagnostic testing.

    Science.gov (United States)

    Freifeld, Alison G; Simonsen, Kari A; Booth, Christine S; Zhao, Xing; Whitney, Scott E; Karre, Teresa; Iwen, Peter C; Viljoen, Hendrik J

    2012-01-01

    We describe a new method for the rapid diagnosis of Clostridium difficile infection, with stool sample preparation and DNA extraction by heat and physical disruption in a single-use lysis microreactor (LMR), followed by a rapid PCR amplification step. All steps can be accomplished in stool samples with discordant EIA results (GDH(+)/toxin(-)) were tested by both the LMR/PCR assay and the loop-mediated isothermal amplification test (LAMP) (Illumigene C. difficile; Meridian Bioscience, Cincinnati, OH). In 64/69 EIA-discordant samples, LAMP and LMR/PCR results matched (both positive in 29 sample and both negative in 35 samples); in the remaining 5 samples, results were discrepant between the LAMP assay (all five negative) and the LMR/PCR assay (all 5 positive). Overall, LMR/PCR testing matched the current algorithm of EIA and/or LAMP reflex testing in 193/198 (97.5%) samples. The present proof-of-concept study suggests that the novel LMR/PCR technique described here may be developed as an inexpensive, rapid, and reliable point-of-care diagnostic test for C. difficile infection and other infectious diseases. Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

  8. Methods of solving rapid binding target-mediated drug disposition model for two drugs competing for the same receptor.

    Science.gov (United States)

    Yan, Xiaoyu; Chen, Yang; Krzyzanski, Wojciech

    2012-10-01

    The target-mediated drug disposition (TMDD) model has been adopted to describe pharmacokinetics for two drugs competing for the same receptor. A rapid binding assumption introduces total receptor and total drug concentrations while free drug concentrations C (A) and C (B) are calculated from the equilibrium (Gaddum) equations. The Gaddum equations are polynomials in C (A) and C (B) of second degree that have explicit solutions involving complex numbers. The aim of this study was to develop numerical methods to solve the rapid binding TMDD model for two drugs competing for the same receptor that can be implemented in pharmacokinetic software. Algebra, calculus, and computer simulations were used to develop algorithms and investigate properties of solutions to the TMDD model with two drugs competitively binding to the same receptor. A general rapid binding approximation of the TMDD model for two drugs competing for the same receptor has been proposed. The explicit solutions to the equilibrium equations employ complex numbers, which cannot be easily solved by pharmacokinetic software. Numerical bisection algorithm and differential representation were developed to solve the system instead of obtaining an explicit solution. The numerical solutions were validated by MATLAB 7.2 solver for polynomial roots. The applicability of these algorithms was demonstrated by simulating concentration-time profiles resulting from exogenous and endogenous IgG competing for the neonatal Fc receptor (FcRn), and darbepoetin competing with endogenous erythropoietin for the erythropoietin receptor. These models were implemented in ADAPT 5 and Phoenix WinNonlin 6.0, respectively.

  9. A Mixed L2 Norm Regularized HRF Estimation Method for Rapid Event-Related fMRI Experiments

    Directory of Open Access Journals (Sweden)

    Yu Lei

    2013-01-01

    Full Text Available Brain state decoding or “mind reading” via multivoxel pattern analysis (MVPA has become a popular focus of functional magnetic resonance imaging (fMRI studies. In brain decoding, stimulus presentation rate is increased as fast as possible to collect many training samples and obtain an effective and reliable classifier or computational model. However, for extremely rapid event-related experiments, the blood-oxygen-level-dependent (BOLD signals evoked by adjacent trials are heavily overlapped in the time domain. Thus, identifying trial-specific BOLD responses is difficult. In addition, voxel-specific hemodynamic response function (HRF, which is useful in MVPA, should be used in estimation to decrease the loss of weak information across voxels and obtain fine-grained spatial information. Regularization methods have been widely used to increase the efficiency of HRF estimates. In this study, we propose a regularization framework called mixed L2 norm regularization. This framework involves Tikhonov regularization and an additional L2 norm regularization term to calculate reliable HRF estimates. This technique improves the accuracy of HRF estimates and significantly increases the classification accuracy of the brain decoding task when applied to a rapid event-related four-category object classification experiment. At last, some essential issues such as the impact of low-frequency fluctuation (LFF and the influence of smoothing are discussed for rapid event-related experiments.

  10. A rapid non-destructive method for quantification of fungal infection on barley and malt

    DEFF Research Database (Denmark)

    Bodevin, Sabrina; Larsen, Tone Glarborg; Lok, Finn

    Description of topic: Barley harvest 2007 in Europe has seen the resurgence of crops highly infected by filamentous fungi. Hence the evaluation of fungal infection on barley and malt kernels by conventional optical measurement methods lacks accuracy and can be tedious. Here we are presenting a new...... camera. The digital image is acquired in less than 10 seconds, over an 18 bands spanning ranging from the ultra-blue (430 nm) to the near-infrared (970 nm) and then analyzed by the VideometerLab® statistical analysis software. Materials and methods for data collection: Sixty European barley samples from...... of Fusarium and hydrophobins were selected and highly infected kernels, artificially inoculated with F. culmorum or F. graminearum, were produced. Results: A good correlation between competitive ELISA tests, real-time PCR experiments and VideometerLab® prediction was found (R2=0.75). The results obtained...

  11. A Rapid and Quantitative Flow Cytometry Method for the Analysis of Membrane Disruptive Antimicrobial Activity.

    Directory of Open Access Journals (Sweden)

    Neil M O'Brien-Simpson

    Full Text Available We describe a microbial flow cytometry method that quantifies within 3 hours antimicrobial peptide (AMP activity, termed Minimum Membrane Disruptive Concentration (MDC. Increasing peptide concentration positively correlates with the extent of bacterial membrane disruption and the calculated MDC is equivalent to its MBC. The activity of AMPs representing three different membranolytic modes of action could be determined for a range of Gram positive and negative bacteria, including the ESKAPE pathogens, E. coli and MRSA. By using the MDC50 concentration of the parent AMP, the method provides high-throughput, quantitative screening of AMP analogues. A unique feature of the MDC assay is that it directly measures peptide/bacteria interactions and lysed cell numbers rather than bacteria survival as with MIC and MBC assays. With the threat of multi-drug resistant bacteria, this high-throughput MDC assay has the potential to aid in the development of novel antimicrobials that target bacteria with improved efficacy.

  12. Loop-mediated isothermal amplification (LAMP)-based method for rapid mushroom species identification.

    Science.gov (United States)

    Vaagt, Franziska; Haase, Ilka; Fischer, Markus

    2013-02-27

    Toxic mushroom species, such as the death cap ( Amanita phalloides ), are responsible for most mushroom poisonings. In the present work, novel loop-mediated isothermal amplification (LAMP) assays were used for the differentiation of even closely related edible and toxic mushroom species. The applicability of these methods was tested by cross-reaction studies and analysis of spiked mushroom samples (raw and fried material). Contaminations at the level of 2% (w/w) could be detected in different mushroom blends. Three detection methods were used: agarose gel analysis, fluorimetric real-time detection, and visual detection by lateral flow dipsticks (LFD). The LAMP assay combined with LFD detection allows the identification of A. phalloides in about 2 h (including DNA extraction) at a very low level of technical equipment (micropestle, water bath, and mobile centrifuge), which makes this technique perfectly suited for on-site applications.

  13. A rapid and simple method for identifying bacterial polar lipid components in wet biomass.

    Science.gov (United States)

    Nguyen, Tuan Manh; Kim, Jaisoo

    2017-08-01

    There are marked differences between wet and freeze-dried cells with regard to the identification of polar lipid components. The determination of the polar lipid composition of freeze-dried cells is well established. However, several approaches to identifying polar lipid components in wet cells have met with limited success owing to the presence of non-polar compounds in the extracts, resulting in a lipid composition with a narrow scope. In this study, we surveyed the lipid profiles of the wet biomasses of three Gram-positive (Microbacterium lacticum, Rhodococcus koreensis, and Streptomyces longwoodensis) and two Gram-negative (Pseudomonas aeruginosa and Novosphingobium capsulatum) bacteria; the results were comparable in quality to those obtained using a standard freeze-dried approach. Moreover, our improved method ensures simple lipid extraction. Overall, the results of the analysis showed minor lipid profile differences between the two approaches with regard to quantity, and lipid identification was consistent in both methods for all species.

  14. The MESERAN Method: Rapid Quantification of Non-Volatile Organic Residue (NVOR)

    Energy Technology Data Exchange (ETDEWEB)

    Benkovich, M.G.

    2002-06-13

    The precision analytical technique known as MESERAN Analysis permits quantitative measurement of the level of preexisting nonvolatile organic residue (NVOR) on a substrate from <1 nanogram (ng)/cm{sup 2} to > 100 micrograms ({micro}g)/cm{sup 2} in 2 minutes. MESERAN Analysis is also applicable to determining NVOR in solvents and solvent extracts. The MESERAN method is able to quantify organic contamination levels down to and below 1 ng by depositing as little as 10 microliters ({micro}L) of solvent containing a known amount of contamination on a clean substrate, allowing it to evaporate, and measuring the evaporated residue. The method will be described in detail and NVOR measurements determined from MESERAN data will be presented.

  15. Rapid continuous chemical methods for studies of nuclei far from stability

    CERN Document Server

    Trautmann, N; Eriksen, D; Gaggeler, H; Greulich, N; Hickmann, U; Kaffrell, N; Skarnemark, G; Stender, E; Zendel, M

    1981-01-01

    Fast continuous separation methods accomplished by combining a gas-jet recoil-transport system with a variety of chemical systems are described. Procedures for the isolation of individual elements from fission product mixtures with the multistage solvent extraction facility SISAK are presented. Thermochromatography in connection with a gas-jet has been studied as a technique for on-line separation of volatile fission halides. Based on chemical reactions in a gas-jet system itself separation procedures for tellurium, selenium and germanium from fission products have been worked out. All the continuous chemical methods can be performed within a few seconds. The application of such procedures to the investigation of nuclides far from the line of beta -stability is illustrated by a few examples. (16 refs).

  16. Accurate and rapid optical characterization of an anisotropic guided structure based on a neural method.

    Science.gov (United States)

    Robert, Stéphane; Battie, Yann; Jamon, Damien; Royer, Francois

    2007-04-10

    Optimal performances of integrated optical devices are obtained by the use of an accurate and reliable characterization method. The parameters of interest, i.e., optical indices and thickness of the waveguide structure, are calculated from effective indices by means of an inversion procedure. We demonstrate how an artificial neural network can achieve such a process. The artificial neural network used is a multilayer perceptron. The first result concerns a simulated anisotropic waveguide. The accuracy in the determination of optical indices and waveguide thickness is 5 x 10(-5) and 4 nm, respectively. Then an experimental application on a silica-titania thin film is performed. In addition, effective indices are measured by m-lines spectroscopy. Finally, a comparison with a classical optimization algorithm demonstrates the robustness of the neural method.

  17. RAPID COMMUNICATION: A TALIF calibration method for quantitative oxygen atom density measurement in plasma jets

    Science.gov (United States)

    Dilecce, G.; Vigliotti, M.; DeBenedictis, S.

    2000-03-01

    In this communication we propose a calibration method for two-photon absorption laser induced fluorescence (TALIF). It can be carried out without any addition or modification to the O atom TALIF set-up. It is based on the measurement of the collision quenching of the laser-excited state (3p3 P2 ) in a pure O2 system in which a high dissociation degree can be achieved. Since the collision rate constant by O is largely lower than that by O2 , the quenching rate can be correlated to the O density. The incertitude in this procedure is comparable to other calibration techniques. We have applied this method to the spatially resolved measurement of O atom density in an O2 rf plasma jet.

  18. Rapid sex identification method of spinach (Spinacia oleracea L.) in the vegetative stage using loop-mediated isothermal amplification.

    Science.gov (United States)

    Fujita, Naoko; Ayukawa, Yu; Fuke, Mitsutoshi; Teraoka, Tohru; Watanabe, Kyoko; Arie, Tsutomu; Komatsu, Ken

    2017-01-01

    A LAMP-mediated, simple and rapid method for sex identification in spinach was developed. Nutrient compositional analysis showed a higher iron content in male than female plants. Spinach (Spinacia oleracea L.) is a dioecious plant with its sex determined by the XY system. Male and female floral organs differ morphologically, but plants do not differ in the vegetative stage before flowering. PCR with Y chromosome markers has been used to determine the sex of dioecious plants before flowering. In this study, we developed a genotype-specific loop-mediated isothermal amplification (LAMP) for sex identification of individual vegetative-stage spinach plants, using primers designed for the genomic region flanked by male-specific markers. LAMP could specifically detect spinach males. The method was further modified to omit DNA purification and use just an aliquot of crude leaf extract homogenized in water. We compared the nutrient composition of males and females, finding higher amounts of iron in the males. Our method could therefore be used for rapidly discriminating male plants in the field, which is useful for efficient hybrid breeding.

  19. A Rapid Detection Method of Brucella with Quantum Dots and Magnetic Beads Conjugated with Different Polyclonal Antibodies

    Science.gov (United States)

    Song, Dandan; Qu, Xiaofeng; Liu, Yushen; Li, Li; Yin, Dehui; Li, Juan; Xu, Kun; Xie, Renguo; Zhai, Yue; Zhang, Huiwen; Bao, Hao; Zhao, Chao; Wang, Juan; Song, Xiuling; Song, Wenzhi

    2017-03-01

    Brucella spp. are facultative intracellular bacteria that cause zoonotic disease of brucellosis worldwide. Traditional methods for detection of Brucella spp. take 48-72 h that does not meet the need of rapid detection. Herein, a new rapid detection method of Brucella was developed based on polyclonal antibody-conjugating quantum dots and antibody-modified magnetic beads. First, polyclonal antibodies IgG and IgY were prepared and then the antibody conjugated with quantum dots (QDs) and immunomagnetic beads (IMB), respectively, which were activated by N-(3-dimethylaminopropyl)- N'-ethylcar-bodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) to form probes. We used the IMB probe to separate the Brucella and labeled by the QD probe, and then detected the fluorescence intensity with a fluorescence spectrometer. The detection method takes 105 min with a limit of detection of 103 CFU/mL and ranges from 10 to 105 CFU/mL ( R 2 = 0.9983), and it can be well used in real samples.

  20. A rapid quantitative method for the analysis of synthetic cannabinoids by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Freijo, Tom D; Harris, Steve E; Kala, Subbarao V

    2014-10-01

    Synthetic cannabinoids represent an emerging drug problem in the USA, as these compounds are constantly being modified and rapidly sold as soon as they become available. Laboratories around the world are constantly improving the analytical methods to detect and identify these newly available designer drugs. This study used a simple approach to detect and quantify a variety of synthetic cannabinoids (14 parent compounds and 15 metabolites including series XLR, AM, JWH, UR, RCS, PB, HU and AB-FUBINACA) using LC-MS-MS. Drug-free urine samples spiked with various synthetic cannabinoids and their metabolites were separated on a C18-Hypersil Gold column using an Agilent 1290 ultra-high performance liquid chromatography and detected by an AB Sciex API 4000 tandem mass spectrometer. Studies were carried out to determine limit of detection, limit of quantitation, upper limit of linearity, ion suppression, interference, precision and accuracy to validate the method. Urine samples from patients and known users were hydrolyzed with β-glucuronidase prior to the analysis by LC-MS-MS, and the data are presented. The method described here is rapid, highly sensitive and specific for the identification of a variety of synthetic cannabinoids. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  1. Rapid resolution of chronic shoulder pain classified as derangement using the McKenzie method: a case series.

    Science.gov (United States)

    Aytona, Maria Corazon; Dudley, Karlene

    2013-11-01

    The McKenzie method, also known as Mechanical Diagnosis and Therapy (MDT), is primarily recognized as an evaluation and treatment method for the spine. However, McKenzie suggested that this method could also be applied to the extremities. Derangement is an MDT classification defined as an anatomical disturbance in the normal resting position of the joint, and McKenzie proposed that repeated movements could be applied to reduce internal joint displacement and rapidly reduce derangement symptoms. However, the current literature on MDT application to shoulder disorders is limited. Here, we present a case series involving four patients with chronic shoulder pain from a duration of 2-18 months classified as derangement and treated using MDT principles. Each patient underwent mechanical assessment and was treated with repeated movements based on their directional preference. All patients demonstrated rapid and clinically significant improvement in baseline measures and the disabilities of the arm, shoulder, and hand (QuickDASH) scores from an average of 38% at initial evaluation to 5% at discharge within 3-5 visits. Our findings suggest that MDT may be an effective treatment approach for shoulder pain.

  2. Rapid Method for the Determination of 5-Hydroxymethylfurfural and Levulinic Acid Using a Double-Wavelength UV Spectroscopy

    Directory of Open Access Journals (Sweden)

    Junhua Zhang

    2013-01-01

    Full Text Available This study reports on a rapid method for the determination of levulinic acid (LA and 5-hydroxymethylfurfural (HMF in acid hydrolyze system of glucose based on UV spectroscopy. It was found that HMF and LA have a maximum absorption at the wavelengths of 284 nm and 266 nm, respectively, in a water medium, and the absorptions of HMF and LA at 284 nm and 266 nm follow Beer’s law very well. However, it was found that a major spectral interference species will arise in the quantification of HMF and LA; nonetheless, this interference can be eliminated through the absorption treatment of charcoal. Therefore, both HMF and LA can be quantified with a double-wavelength technique. The repeatability of the method had a relative standard deviation of less than 4.47% for HMF and 2.25% for LA; the limit of quantification (LOQ was 0.017 mmol/L for HMF and 4.68 mmol/L for LA, and the recovery ranged from 88% to 116% for HMF and from 94% to 105% for LA. The present method is simple, rapid, and accurate. It is suitable to use in the research of the preparation of HMF and LA in biorefinery area.

  3. DPS - a rapid method for genome sequencing of DNA-containing bacteriophages directly from a single plaque.

    Science.gov (United States)

    Kot, Witold; Vogensen, Finn K; Sørensen, Søren J; Hansen, Lars H

    2014-02-01

    Bacteriophages (phages) coexist with bacteria in all environments and influence microbial diversity, evolution and industrial production processes. As a result of this major impact of phages on microbes, tools that allow rapid characterization of phages are needed. Today, one of the most powerful methods for characterization of phages is determination of the whole genome using high throughput sequencing approaches. Here a direct plaque sequencing (DPS) is described, which is a rapid method that allows easy full genome sequencing of DNA-containing phages using the Nextera XT™ kit. A combination of host-DNA removal followed by purification and concentration of the viral DNA, allowed the construction of Illumina-compatible sequencing libraries using the Nextera™ XT technology directly from single phage plaques without any whole genome amplification step. This method was tested on three Caudovirales phages; ϕ29 Podoviridae, P113g Siphoviridae and T4 Myovirdae, which are representative of >96% of all known phages, and were sequenced using the Illumina MiSeq platform. Successful de novo assembly of the viral genomes was possible. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Method Optimization for Rapid Measurement of Carbohydrates in Plasma by Liquid Chromatography Tandem Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Ductoan; Yu, Jondong; Mho, Sunil; Lee, Gwang [Ajou Univ., Suwon (Korea, Republic of); Lee, Haelee; Paik, Manjeong [Sangdosijang Pharmacy, Seoul (Korea, Republic of); Yee, Sungtae [Sunchon National Univ., Suncheon (Korea, Republic of)

    2013-05-15

    In conclusion, the developed HPLC coupled with ESI-MS was a powerful technique for the separation and characterization of carbohydrates by either SIM or MRM mode. The present method will be useful for the monitoring of carbohydrate profile in biological fluids from various diseases including diabetic ketoacidosis, hypoglycemia and hyperosmolar coma. Carbohydrates are one of the most abundant classes of organic compounds in nature, which not only constitute complex biomolecules in human and animals but are also distributed in plants and bacteria.

  5. A simple and rapid method to characterize lipid fate in skeletal muscle

    OpenAIRE

    Massart, Julie; Zierath, Juleen R; Chibalin, Alexander V

    2014-01-01

    Background Elevated fatty acids contribute to the development of type 2 diabetes and affect skeletal muscle insulin sensitivity. Since elevated intramuscular lipids and insulin resistance is strongly correlated, aberrant lipid storage or lipid intermediates may be involved in diabetes pathogenesis. The aim of this study was to develop a method to determine the dynamic metabolic fate of lipids in primary human skeletal muscle cells and in intact mouse skeletal muscle. We report a simple and fa...

  6. Bed Evolution under Rapidly Varying Flows by a New Method for Wave Speed Estimation

    Directory of Open Access Journals (Sweden)

    Khawar Rehman

    2016-05-01

    Full Text Available This paper proposes a sediment-transport model based on coupled Saint-Venant and Exner equations. A finite volume method of Godunov type with predictor-corrector steps is used to solve a set of coupled equations. An efficient combination of approximate Riemann solvers is proposed to compute fluxes associated with sediment-laden flow. In addition, a new method is proposed for computing the water depth and velocity values along the shear wave. This method ensures smooth solutions, even for flows with high discontinuities, and on domains with highly distorted grids. The numerical model is tested for channel aggradation on a sloping bottom, dam-break cases at flume-scale and reach-scale with flat bottom configurations and varying downstream water depths. The proposed model is tested for predicting the position of hydraulic jump, wave front propagation, and for predicting magnitude of bed erosion. The comparison between results based on the proposed scheme and analytical, experimental, and published numerical results shows good agreement. Sensitivity analysis shows that the model is computationally efficient and virtually independent of mesh refinement.

  7. A simple and rapid method for high-resolution visualization of single-ion tracks

    Energy Technology Data Exchange (ETDEWEB)

    Omichi, Masaaki [Department of Applied Chemistry, Graduate School of Engineering, Osaka University, Osaka 565-0871 (Japan); Center for Collaborative Research, Anan National College of Technology, Anan, Tokushima 774-0017 (Japan); Choi, Wookjin; Sakamaki, Daisuke; Seki, Shu, E-mail: seki@chem.eng.osaka-u.ac.jp [Department of Applied Chemistry, Graduate School of Engineering, Osaka University, Osaka 565-0871 (Japan); Tsukuda, Satoshi [Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Miyagi 980-8577 (Japan); Sugimoto, Masaki [Japan Atomic Energy Agency, Takasaki Advanced Radiation Research Institute, Gunma, Gunma 370-1292 (Japan)

    2014-11-15

    Prompt determination of spatial points of single-ion tracks plays a key role in high-energy particle induced-cancer therapy and gene/plant mutations. In this study, a simple method for the high-resolution visualization of single-ion tracks without etching was developed through the use of polyacrylic acid (PAA)-N, N’-methylene bisacrylamide (MBAAm) blend films. One of the steps of the proposed method includes exposure of the irradiated films to water vapor for several minutes. Water vapor was found to promote the cross-linking reaction of PAA and MBAAm to form a bulky cross-linked structure; the ion-track scars were detectable at a nanometer scale by atomic force microscopy. This study demonstrated that each scar is easily distinguishable, and the amount of generated radicals of the ion tracks can be estimated by measuring the height of the scars, even in highly dense ion tracks. This method is suitable for the visualization of the penumbra region in a single-ion track with a high spatial resolution of 50 nm, which is sufficiently small to confirm that a single ion hits a cell nucleus with a size ranging between 5 and 20 μm.

  8. A simple and rapid method for high-resolution visualization of single-ion tracks

    Directory of Open Access Journals (Sweden)

    Masaaki Omichi

    2014-11-01

    Full Text Available Prompt determination of spatial points of single-ion tracks plays a key role in high-energy particle induced-cancer therapy and gene/plant mutations. In this study, a simple method for the high-resolution visualization of single-ion tracks without etching was developed through the use of polyacrylic acid (PAA-N, N’-methylene bisacrylamide (MBAAm blend films. One of the steps of the proposed method includes exposure of the irradiated films to water vapor for several minutes. Water vapor was found to promote the cross-linking reaction of PAA and MBAAm to form a bulky cross-linked structure; the ion-track scars were detectable at a nanometer scale by atomic force microscopy. This study demonstrated that each scar is easily distinguishable, and the amount of generated radicals of the ion tracks can be estimated by measuring the height of the scars, even in highly dense ion tracks. This method is suitable for the visualization of the penumbra region in a single-ion track with a high spatial resolution of 50 nm, which is sufficiently small to confirm that a single ion hits a cell nucleus with a size ranging between 5 and 20 μm.

  9. Microplate-reader method for the rapid analysis of copper in natural waters with chemiluminescence detection

    Directory of Open Access Journals (Sweden)

    Axel eDurand

    2013-01-01

    Full Text Available We have developed a method for the determination of copper in natural waters at nanomolar levels. The use of a microplate-reader minimises sample processing time (~ 25 sec per sample, reagent consumption (~ 120 μL per sample and sample volume (~ 700 μL. Copper is detected by chemiluminescence. This technique is based on the formation of a complex between copper and 1,10-phenanthroline and the subsequent emission of light during the oxidation of the complex by hydrogen peroxide. Samples are acidified to pH 1.7 and then introduced directly into a 24-well plate. Reagents are added during data acquisition via two reagent injectors. When trace metal clean protocols are employed, the reproducibility is generally less then 7% on blanks and the detection limit is 0.7 nM for seawater and 0.4 nM for freshwater. More than 100 samples per hour can be analyzed with this technique, which is simple, robust, and amenable to at-sea analysis. Seawater samples from Storm Bay in Tasmania illustrate the utility of the method for environmental science. Indeed other trace metals for which optical detection methods exist (e.g. chemiluminescence, fluorescence and absorbance could be adapted to the microplate-reader.

  10. Human serum processing and analysis methods for rapid and reproducible N-glycan mass profiling.

    Science.gov (United States)

    Kronewitter, Scott R; de Leoz, Maria Lorna A; Peacock, Kyle S; McBride, Kelly R; An, Hyun Joo; Miyamoto, Suzanne; Leiserowitz, Gary S; Lebrilla, Carlito B

    2010-10-01

    Glycans constitute a new class of compounds for biomarker discovery. Glycosylation is a common post-translational modification and is often associated with transformation to malignancy. To analyze glycans, they are released from proteins, enriched, and measured with mass spectrometry. For biomarker discovery, repeatability at every step of the process is important. Locating and minimizing the process variability is key to establishing a robust platform stable enough for biomarker discovery. Understanding the variability of the measurement devices helps understand the variability associated with the chemical processing. This report explores the potential use of methods expediting the enzymatic release of glycans such as a microwave reactor and automation of the solid-phase extraction with a robotic liquid handler. The study employs matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry but would be suitable with any mass spectrometry method. Methods for system-wide data analysis are examined because proper metrics for evaluating the performance of glycan sample preparation procedures are not well established.

  11. A simple and rapid resonance Rayleigh scattering method for detection of indigo carmine in soft drink.

    Science.gov (United States)

    Li, Qin; Yang, Jidong; Tan, Xuanping; Zhang, Zhan; Hu, Xiaomei; Yang, Menghuan

    2016-08-01

    A novel method that uses acridine orange (AO) to detect indigo carmine (IC) in soft drinks was developed. The method is highly sensitive and is based on a resonance Rayleigh scattering (RRS) technique. In Britton-Robinson (BR) buffer solution, pH 4.3, the weak RRS intensity of AO was greatly enhanced by the addition of IC, with the maximum peak located at 332 nm. Under optimum conditions, it was found that the enhanced RRS intensity was proportional to the concentration of IC over a range of 2-32 × 10(-6)  mol/L. A low detection limit of 2.4 × 10(-8)  mol/L was achieved. The sensitivity and selectivity of the method are high enough to permit the determination of trace amounts of IC without any significant interference from high levels of other components such as common anions and other amino acids. Finally, the concentration of IC in three different soft drinks was determined with satisfactory results. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  12. Novel Simplified and Rapid Method for Screening and Isolation of Polyunsaturated Fatty Acids Producing Marine Bacteria

    Directory of Open Access Journals (Sweden)

    Ashwini Tilay

    2012-01-01

    Full Text Available Bacterial production of polyunsaturated fatty acids (PUFAs is a potential biotechnological approach for production of valuable nutraceuticals. Reliable method for screening of number of strains within short period of time is great need. Here, we report a novel simplified method for screening and isolation of PUFA-producing bacteria by direct visualization using the H2O2-plate assay. The oxidative stability of PUFAs in growing bacteria towards added H2O2 is a distinguishing characteristic between the PUFAs producers (no zone of inhibition and non-PUFAs producers (zone of inhibition by direct visualization. The confirmation of assay results was performed by injecting fatty acid methyl esters (FAMEs produced by selected marine bacteria to Gas Chromatography-Mass Spectrometry (GCMS. To date, this assay is the most effective, inexpensive, and specific method for bacteria producing PUFAs and shows drastically reduction in the number of samples thus saves the time, effort, and cost of screening and isolating strains of bacterial PUFAs producers.

  13. A rapid method for alpha-spectrometric analysis of radium isotopes in natural waters using ion-selective membrane technology.

    Science.gov (United States)

    Purkl, Stefan; Eisenhauer, Anton

    2003-10-01

    An alpha-spectrometric method for the rapid determination of radium isotopes (223Ra, 224Ra and 226Ra) in environmental samples is presented. Using Empore Radium Rad Disks complete separation of the target radionuclides is achieved. The high selectivity of these Rad Disks allows the straightforward use of 225Ra as yield tracer. Chemical yield is up to 92 +/- 9%. The chemical procedure can be accomplished within 5h. Alpha-sources show energy resolution in the range of typically 26-40 keV (FWHM). Despite minimal thickness of the sources no significant radon (Rn) losses can be observed.

  14. Evaluation of usefulness of single-strand conformation polymorphism method for rapid detection of rifampicin-resistant mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Tešić Jelena

    2004-01-01

    Full Text Available The aim was to evaluate the Single-Strand Conformation Polymorphism method as a potential tool for rapid detection of rifampicin-resistant strains by the use of 39 rifampicin-resistant Mycobacterium tuberculosis strains isolated in Serbia. SSCP analysis on acrylamide gel detected 56.4% of the rifampicin-resistant M. tuberculosis strains and showed the inability to detect one of the most frequent mutations, TCG®TTG mutation in codon 531 of the rpoB gene, which was shown by automated sequencing.

  15. Rapid detection of Salmonella in pet food: design and evaluation of integrated methods based on real-time PCR detection.

    Science.gov (United States)

    Balachandran, Priya; Friberg, Maria; Vanlandingham, V; Kozak, K; Manolis, Amanda; Brevnov, Maxim; Crowley, Erin; Bird, Patrick; Goins, David; Furtado, Manohar R; Petrauskene, Olga V; Tebbs, Robert S; Charbonneau, Duane

    2012-02-01

    Reducing the risk of Salmonella contamination in pet food is critical for both companion animals and humans, and its importance is reflected by the substantial increase in the demand for pathogen testing. Accurate and rapid detection of foodborne pathogens improves food safety, protects the public health, and benefits food producers by assuring product quality while facilitating product release in a timely manner. Traditional culture-based methods for Salmonella screening are laborious and can take 5 to 7 days to obtain definitive results. In this study, we developed two methods for the detection of low levels of Salmonella in pet food using real-time PCR: (i) detection of Salmonella in 25 g of dried pet food in less than 14 h with an automated magnetic bead-based nucleic acid extraction method and (ii) detection of Salmonella in 375 g of composite dry pet food matrix in less than 24 h with a manual centrifugation-based nucleic acid preparation method. Both methods included a preclarification step using a novel protocol that removes food matrix-associated debris and PCR inhibitors and improves the sensitivity of detection. Validation studies revealed no significant differences between the two real-time PCR methods and the standard U.S. Food and Drug Administration Bacteriological Analytical Manual (chapter 5) culture confirmation method.

  16. Chemical Screening Method for the Rapid Identification of Microbial Sources of Marine Invertebrate-Associated Metabolites

    Directory of Open Access Journals (Sweden)

    Russell G. Kerr

    2011-03-01

    Full Text Available Marine invertebrates have proven to be a rich source of secondary metabolites. The growing recognition that marine microorganisms associated with invertebrate hosts are involved in the biosynthesis of secondary metabolites offers new alternatives for the discovery and development of marine natural products. However, the discovery of microorganisms producing secondary metabolites previously attributed to an invertebrate host poses a significant challenge. This study describes an efficient chemical screening method utilizing a 96-well plate-based bacterial cultivation strategy to identify and isolate microbial producers of marine invertebrate-associated metabolites.

  17. Rapid multiplex high resolution melting method to analyze inflammatory related SNPs in preterm birth

    Directory of Open Access Journals (Sweden)

    Pereyra Silvana

    2012-01-01

    Full Text Available Abstract Background Complex traits like cancer, diabetes, obesity or schizophrenia arise from an intricate interaction between genetic and environmental factors. Complex disorders often cluster in families without a clear-cut pattern of inheritance. Genomic wide association studies focus on the detection of tens or hundreds individual markers contributing to complex diseases. In order to test if a subset of single nucleotide polymorphisms (SNPs from candidate genes are associated to a condition of interest in a particular individual or group of people, new techniques are needed. High-resolution melting (HRM analysis is a new method in which polymerase chain reaction (PCR and mutations scanning are carried out simultaneously in a closed tube, making the procedure fast, inexpensive and easy. Preterm birth (PTB is considered a complex disease, where genetic and environmental factors interact to carry out the delivery of a newborn before 37 weeks of gestation. It is accepted that inflammation plays an important role in pregnancy and PTB. Methods Here, we used real time-PCR followed by HRM analysis to simultaneously identify several gene variations involved in inflammatory pathways on preterm labor. SNPs from TLR4, IL6, IL1 beta and IL12RB genes were analyzed in a case-control study. The results were confirmed either by sequencing or by PCR followed by restriction fragment length polymorphism. Results We were able to simultaneously recognize the variations of four genes with similar accuracy than other methods. In order to obtain non-overlapping melting temperatures, the key step in this strategy was primer design. Genotypic frequencies found for each SNP are in concordance with those previously described in similar populations. None of the studied SNPs were associated with PTB. Conclusions Several gene variations related to the same inflammatory pathway were screened through a new flexible, fast and non expensive method with the purpose of analyzing

  18. Multithermal titration calorimetry: a rapid method to determine binding heat capacities.

    Science.gov (United States)

    Chavelas, Eneas A; Zubillaga, Rafael A; Pulido, Nancy O; García-Hernández, Enrique

    2006-03-01

    Herein a new method that allows binding DeltaCp to be determined with a single experiment is presented. Multithermal titration calorimetry (MTC) is a simple extension of isothermal titration calorimetry (ITC) that explicitly takes into account the thermal dependences of DeltaH and the binding constant. Experimentally, this is accomplished by performing a single stepwise titration with ITC equipment, allowing temperature re-adjustments of the system at intermediate states of the titration process. Thus, from the resulting multitherm, DeltaCp can also be determined. The experimental feasibility of MTC was tested by using the well-characterized lysozyme-chitotriose complex as a model system.

  19. Measurement of rapid membrane permeation in cell suspensions by application of a generalized capillary method

    DEFF Research Database (Denmark)

    Klösgen, Beate; Schönert, Hansjürgen; Deuticke, Bernhard

    1988-01-01

    the diffusion process of a solute in a composite system was derived using a series-parallel-pathway model with explicit consideration of the diffusion pathways inside and between the cells. This renders the technique insensitive to unstirred layer effects. Any single cell population of known size distribution...... may be investigated. High permeabilities (above 5 · 10-3 cm/s) can be measured with the greatest precision, but lower permeabilities, down to a limit of about 5 · 10-4 cm/s, may also be determined by the method. Measurements in erythrocyte suspensions have been made using non...... of suspensions of membrane vesicles....

  20. A neurophysiological method of rapid detection and analysis of marine algal toxins

    DEFF Research Database (Denmark)

    Kerr, DS; Bødtkjer, Donna Briggs; Saba, HI

    1999-01-01

    on either antidromic or fibre spikes. Fifty nanomolar saxitoxin (PSP) abolished all responses in all slices. Only antidromic spikes showed any recovery during wash-out. Field EPSP and fiber spike analysis further demonstrated that the preparation is capable of reliably detecting saxitoxin in a linearly...... responsive fashion at toxin concentrations of 25-200 nM, and tests of naturally contaminated shellfish confirmed the utility of this assay as a screening method for PSP. Our findings suggest that the in vitro hippocampal slice preparation has potential in the detection and analysis of three marine algal...

  1. A rapid detection method for PAI-1 promoter insertion/deletion polymorphism (4G/5G

    Directory of Open Access Journals (Sweden)

    Annichino-Bizzacchi Joyce M.

    1998-01-01

    Full Text Available Plasminogen activator inhibitor-1 (PAI-1 is an important inhibitor of fibrinolysis, and increased levels of PAI-1 are associated with atheroma and myocardial infarction. A common 4G/5G insertion/deletion polymorphism located in the promoter region of PAI-1 gene has been described associated with PAI-1 activity in plasma levels. Genotyping of this polymorphism is commonly conducted with an allele-specific oligonucleotide melting technique. In the present study, we describe a quick, easy method for genotyping 4G/5G polymorphism in the promoter region of the PAI-1 gene.

  2. A rapid method for standardization of the gated radionuclide left ventricular ejection fraction

    Energy Technology Data Exchange (ETDEWEB)

    Purnell, G.L.; Baker, B.J.; Murphy, M.M.; Boyd, C.M.; Dalrymple, G.V. (Univ. of Arkansas for Medical Sciences (USA))

    1990-08-01

    Radionuclide ventriculography has become the standard method for serially evaluating left ventricular ejection fraction (EF). The gold standard for evaluating EF is the contrast ventriculogram, which uses mathematical models to arrive at the volumes used to calculate EF. These models are subject to possible error. This paper reports the standardization of volume measurements of a digital angiographic camera system using a series of cardiac phantoms and the correlation of measurement of the EF of a series of patients whose EF was determined by digital angiography and radionuclide ventriculography.

  3. Evaluation of methods of rapid mass killing of segregated early weaned piglets.

    Science.gov (United States)

    Whiting, Terry L; Steele, Gregory G; Wamnes, Steinar; Green, Chris

    2011-07-01

    The operational logistics of mass killing of healthy, surplus piglets by manual blunt force trauma, controlled blunt force trauma, intraperitoneal injection of barbiturate, and free bullet were recorded. Objective performance variables evaluated were, speed of application, human resource and input cost, animal restraint required, and failure rate. Subjective evaluation of esthetics and difficulty of application indicated manual blunt force trauma is an unacceptable technique. Under field conditions, physical methods of killing were superior to intraperitoneal injection of concentrated pentobarbital. Considering animal welfare metrics in isolation, controlled blunt force trauma was superior to all other techniques attempted.

  4. Method for rapid optimization of recombinant GPCR protein expression and stability using virus-like particles.

    Science.gov (United States)

    Ho, Thao T; Nguyen, Jasmine T; Liu, Juping; Stanczak, Pawel; Thompson, Aaron A; Yan, Yingzhuo G; Chen, Jasmine; Allerston, Charles K; Dillard, Charles L; Xu, Hao; Shoger, Nicholas J; Cameron, Jill S; Massari, Mark E; Aertgeerts, Kathleen

    2017-05-01

    Recent innovative approaches to stabilize and crystallize GPCRs have resulted in an unprecedented breakthrough in GPCR crystal structures as well as application of the purified receptor protein in biophysical and biochemical ligand binding assays. However, the protein optimization process to enable these technologies is lengthy and requires iterative overexpression, solubilization, purification and functional analysis of tens to hundreds of protein variants. Here, we report a new and versatile method to screen in parallel hundreds of GPCR variants in HEK293 produced virus-like particles (VLPs) for protein yield, stability, functionality and ligand binding. This approach reduces the time and resources during GPCR construct optimization by eliminating lengthy protein solubilization and purification steps and by its adaptability to many binding assay formats (label or label-free detection). We exemplified the robustness of our VLP method by screening 210 GALR3-VLP variants in a radiometric agonist-based binding assay and a subset of 88 variants in a label-free antagonist-based assay. The resulting GALR3 agonist or antagonist stabilizing variants were then further used for recombinant protein expression in transfected insect cells. The final purified protein variants were successfully immobilized on a biosensor chip and used in a surface plasmon resonance binding assay. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Colony-PCR Is a Rapid Method for DNA Amplification of Hyphomycetes

    Directory of Open Access Journals (Sweden)

    Georg Walch

    2016-04-01

    Full Text Available Fungal pure cultures identified with both classical morphological methods and through barcoding sequences are a basic requirement for reliable reference sequences in public databases. Improved techniques for an accelerated DNA barcode reference library construction will result in considerably improved sequence databases covering a wider taxonomic range. Fast, cheap, and reliable methods for obtaining DNA sequences from fungal isolates are, therefore, a valuable tool for the scientific community. Direct colony PCR was already successfully established for yeasts, but has not been evaluated for a wide range of anamorphic soil fungi up to now, and a direct amplification protocol for hyphomycetes without tissue pre-treatment has not been published so far. Here, we present a colony PCR technique directly from fungal hyphae without previous DNA extraction or other prior manipulation. Seven hundred eighty-eight fungal strains from 48 genera were tested with a success rate of 86%. PCR success varied considerably: DNA of fungi belonging to the genera Cladosporium, Geomyces, Fusarium, and Mortierella could be amplified with high success. DNA of soil-borne yeasts was always successfully amplified. Absidia, Mucor, Trichoderma, and Penicillium isolates had noticeably lower PCR success.

  6. Simple, rapid method for quantification of tricuspid regurgitation by two-dimensional echocardiography.

    Science.gov (United States)

    Fisher, E A; Goldman, M E

    1989-06-01

    Functional tricuspid regurgitation (TR) is usually due to pulmonary hypertension with subsequent right ventricular and tricuspid valve anular dilatation. The correlation between anular dilatation and TR severity is unclear. Unfortunately, there is no simple, accurate method of assessing the presence and severity of TR. Real-time (color flow) Doppler echocardiography, a sophisticated but expensive technique, facilitates noninvasive determination of the presence and severity of TR by the area and depth of color-encoded, systolic turbulent reflux into the right atrium. In this study, TR severity (0 to 4+), assessed by color flow Doppler, was correlated with diastolic and systolic tricuspid valve anulus dimension, measured by computerized calipers, from the apical 4-chamber and right ventricular inflow 2-dimensional views. Forty-six patients were studied: 32 with 0 to 2+ TR and 14 with functional 3 to 4+ TR. TR severity correlated significantly with the tricuspid valve anulus in both diastole and systole. The correlation was best in the apical 4-chamber view during systole (r = 0.88, p less than 0.0001). No patient with tricuspid valve anulus in systole less than 3.2 cm or in diastole less than 3.4 cm had severe (3 to 4+) TR. Thus, measurement of the tricuspid valve anulus by 2-dimensional echocardiography is a simple, noninvasive method to identify patients with moderate to severe TR.

  7. Field application of a rapid spectrophotometric method for determination of persulfate in soil.

    Directory of Open Access Journals (Sweden)

    Colin J Cunningham

    Full Text Available Remediation of hydrocarbon contaminated soils can be performed both in situ and ex situ using chemical oxidants such as sodium persulfate. Standard methods for quantifying persulfate require either centrifugation or prolonged settling times. An optimized soil extraction procedure was developed for persulfate involving simple water extraction using a modified disposable syringe. This allows considerable saving of time and removes the need for centrifugation. The extraction time was reduced to only 5 min compared to 15 min for the standard approach. A comparison of the two approaches demonstrated that each provides comparable results. Comparisons were made using high (93 g kg(-1 soil and low (9.3 g kg(-1 soil additions of sodium persulfate to a petroleum hydrocarbon-contaminated soil, as well as sand spiked with diesel. Recoveries of 95±1% and 96±10% were observed with the higher application rate in the contaminated soil and spiked sand, respectively. Corresponding recoveries of 86±5% and 117±19% were measured for the lower application rate. Results were obtained in only 25 min and the method is well suited to batch analyses. In addition, it is suitable for application in a small field laboratory or even a mobile, vehicle-based system, as it requires minimal equipment and reagents.

  8. Rapid DNA transformation in Salmonella Typhimurium by the hydrogel exposure method.

    Science.gov (United States)

    Elabed, Hamouda; Hamza, Rim; Bakhrouf, Amina; Gaddour, Kamel

    2016-07-01

    Even with advances in molecular cloning and DNA transformation, new or alternative methods that permit DNA penetration in Salmonella enterica subspecies enterica serovar Typhimurium are required in order to use this pathogen in biotechnological or medical applications. In this work, an adapted protocol of bacterial transformation with plasmid DNA based on the "Yoshida effect" was applied and optimized on Salmonella enterica serovar Typhimurium LT2 reference strain. The plasmid transference based on the use of sepiolite as acicular materials to promote cell piercing via friction forces produced by spreading on the surface of a hydrogel. The transforming mixture containing sepiolite nanofibers, bacterial cells to be transformed and plasmid DNA were plated directly on selective medium containing 2% agar. In order to improve the procedure, three variables were tested and the transformation of Salmonella cells was accomplished using plasmids pUC19 and pBR322. Using the optimized protocol on Salmonella LT2 strain, the efficiency was about 10(5) transformed cells per 10(9) subjected to transformation with 0.2μg plasmid DNA. In summary, the procedure is fast, offers opportune efficiency and promises to become one of the widely used transformation methods in laboratories. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Development of a Rapid and Sensitive HPLC Assay Method for Lenalidomide Capsules and Its Related Substances

    Directory of Open Access Journals (Sweden)

    L. Maheshwara Reddy

    2012-01-01

    Full Text Available A chromatographic method was established for the determination of lenalidomide and related substances in 10 mg and 5 mg capsules using Sunfire C-18(250×4.6 mm ID, 5 μm HPCL column with 85:15 v/v ratio of mobile phases A (mixture of phosphoric acid buffer and 1-octane sulphonic acid sodium salt and B(55: 45 v/v ratio of methanol and acetonitrile at 40°C and 210 nm wave length. The degradation studies were performed using 0.1N HCl, 0.1 N NaOH, 1% v/v hydrogen peroxide, humidity, UV at 254 nm, Sun light, and heat to 60°C. No significant degradation of lenalidomide was observed. However, the slight degradation was observed in presence of NaOH. The developed HPLC method gave the peaks purity angle was less their threshold angle, indicating it to be suitable for stability studies. It was validated with respect to linearity, accuracy, precision, ruggedness, and robustness.

  10. A new method of rapid power measurement for MW-scale high-current particle beams

    Science.gov (United States)

    Xu, Yongjian; Hu, Chundong; Xie, Yuanlai; Liu, Zhimin; Xie, Yahong; Liu, Sheng; Liang, Lizheng; Jiang, Caichao; Sheng, Peng; Yu, Ling

    2015-09-01

    MW-scale high current particle beams are widely applied for plasma heating in the magnetic confinement fusion devices, in which beam power is an important indicator for efficient heating. Generally, power measurement of MW-scale high current particle beam adopts water flow calorimetry (WFC). Limited by the principles of WFC, the beam power given by WFC is an averaged value. In this article a new method of beam power for MW-scale high-current particle beams is introduced: (1) the temperature data of thermocouples embedded in the beam stopping elements were obtained using high data acquire system, (2) the surface heat flux of the beam stopping elements are calculated using heat transfer, (3) the relationships between positions and heat flux were acquired using numerical simulation, (4) the real-time power deposited on the beam stopping elements can be calculated using surface integral. The principle of measurement was described in detail and applied to the EAST neutral beam injector for demonstration. The result is compared with that measured by WFC. Comparison of the results shows good accuracy and applicability of this measuring method.

  11. Longitudinal analysis of the temporal evolution of Acinetobacter baumannii strains in Ohio, USA, by using rapid automated typing methods.

    Directory of Open Access Journals (Sweden)

    Brooke K Decker

    Full Text Available Genotyping methods are essential to understand the transmission dynamics of Acinetobacter baumannii. We examined the representative genotypes of A. baumannii at different time periods in select locations in Ohio, using two rapid automated typing methods: PCR coupled with electrospray ionization mass spectrometry (PCR/ESI-MS, a form of multi-locus sequence typing (MLST, and repetitive-sequence-based-PCR (rep-PCR. Our analysis included 122 isolates from 4 referral hospital systems, in 2 urban areas of Ohio. These isolates were associated with outbreaks at 3 different time periods (1996, 2000 and 2005-2007. Type assignments of PCR/ESI-MS and rep-PCR were compared to each other and to worldwide (WW clone types. The discriminatory power of each method was determined using the Simpson's index of diversity (DI. We observed that PCR/ESI-MS sequence type (ST 14, corresponding to WW clone 3, predominated in 1996, whereas ST 12 and 14 co-existed in the intermediate period (2000 and ST 10 and 12, belonging to WW clone 2, predominated more recently in 2007. The shift from WW clone 3 to WW clone 2 was accompanied by an increase in carbapenem resistance. The DI was approximately 0.74 for PCR/ESI-MS, 0.88 for rep-PCR and 0.90 for the combination of both typing methods. We conclude that combining rapid automated typing methods such as PCR/ESI-MS and rep-PCR serves to optimally characterize the regional molecular epidemiology of A. baumannii. Our data also sheds light on the changing sequence types in an 11 year period in Northeast Ohio.

  12. Evaluation of X-ray fluorescence spectroscopy as a method for the rapid and direct determination of sodium in cheese.

    Science.gov (United States)

    Stankey, J A; Akbulut, C; Romero, J E; Govindasamy-Lucey, S

    2015-08-01

    Cheese manufacturers indirectly determine Na in cheese by analysis of Cl using the Volhard method, assuming that all Cl came from NaCl. This method overestimates the actual Na content in cheeses when Na replacers (e.g., KCl) are used. A direct and rapid method for Na detection is needed. X-ray fluorescence spectroscopy (XRF), a mineral analysis technique used in the mining industry, was investigated as an alternative method of Na detection in cheese. An XRF method for the detection of Na in cheese was developed and compared with inductively coupled plasma optical emission spectroscopy (ICP-OES; the reference method for Na in cheese) and Cl analyzer. Sodium quantification was performed by multi-point calibration with cheese standards spiked with NaCl ranging from 0 to 4% Na (wt/wt). The Na concentration of each of the cheese standards (discs: 30mm×7mm) was quantified by the 3 methods. A single laboratory method validation was performed; linearity, precision, limit of detection, and limit of quantification were determined. An additional calibration graph was created using cheese standards made from natural or process cheeses manufactured with different ratios of Na:K. Both Na and K calibration curves were linear for the cheese standards. Sodium was quantified in a variety of commercial cheese samples. The Na data obtained by XRF were in agreement with those from ICP-OES and Cl analyzer for most commercial natural cheeses. The XRF method did not accurately determine Na concentration for several process cheese samples, compared with ICP-OES, likely due to the use of unknown types of Na-based emulsifying salts (ES). When a calibration curve was created for process cheese with the specific types of ES used for this cheese, Na content was successfully predicted in the samples. For natural cheeses, the limit of detection and limit of quantification for Na that can be determined with an acceptable level of repeatability, precision, and trueness was 82 and 246mg/100g of

  13. Non-invasive method for rapid bedside estimation of inotropy: theory and preliminary clinical validation.

    Science.gov (United States)

    Smith, B E; Madigan, V M

    2013-10-01

    There are numerous techniques which attempt to quantify inotropy (or myocardial contractility). None has yet found general acceptance in anaesthesia and critical care as a practical method. We report a novel approach to the determination of inotropy as a bedside procedure which could identify low inotropy states in patients with clinical heart failure. We estimated the potential and kinetic energy delivered by the left ventricle using continuous-wave Doppler ultrasound (ultrasonic cardiac output monitor, Uscom, Sydney, Australia) and data available at the point of care. A formula to calculate effective inotropy [Smith-Madigan inotropy index (SMII)] was tested against historical haemodynamic data for 250 control subjects (ASA I patients from preoperative clinic) and 83 patients with acute left ventricular failure (LVF) of New York Heart Association Grade 4 (LVF group). The ratio of potential to kinetic energy (PKR) was investigated as a measure of arterial impedance. Significant differences were found between the control and LVF groups for cardiac index, mean (range)=3.37 (2.84-5.32) vs 1.84 (1.43-2.26) litre min(-1) m(-2); stroke volume index (SVI), 49.2 (39-55) vs 34.3 (23-37) ml m(-2); systemic vascular resistance, 893 (644-1242) vs 1960 (1744-4048) dyn s cm(-5); SMII, 1.78 (1.35-2.24) vs 0.73 (0.43-0.97) W m(-2); and PKR, 29:1 (24-35:1) vs 124:1 (96-174:1), Pcalculated for SMII and PKR as mean (+/-1.96) standard deviations, yielding 1.6-2.2 W m(-2) for SMII, and 25-34:1 for PKR. The method clearly identified the two clinical groups with no overlap of data points. The discriminant power of SMII and PKR may offer valuable diagnostic methods and monitoring tools in anaesthesia and critical care. This is the first report of normal ranges for SMII and PKR.

  14. Electromolecular propulsion (EMP): a rapid, simple method for analyzing dyes used in microscopy.

    Science.gov (United States)

    Haber, N

    1998-03-01

    The electrokinetic molecular effect known as electromolecular propulsion (EMP) was used to examine a variety of commonly used dyes that are used as biological stains. The dyes were electrokinetically mobilized using several organic solvent mixtures on a variety of thin layer substrates. Analysis was completed in seconds to minutes. Nearly all dyes tested separated into multiple colored components. The solvents and substrates used were well suited for qualitatively analyzing a broad variety of hydrophilic and lipophilic colorants. Speed, resolving power and operational simplicity of this technique make it convenient for effectively "fingerprinting" for consistency in dye products. EMP may be broadly applied to study quality control of dyes and their stains. Components not ordinarily known to be present can be readily detected using this technology. EMP can complement or offer a versatile alternative to existing analytical methods.

  15. Rapid and Inexpensive Screening of Genomic Copy Number Variations Using a Novel Quantitative Fluorescent PCR Method

    Directory of Open Access Journals (Sweden)

    Martin Stofanko

    2013-01-01

    Full Text Available Detection of human microdeletion and microduplication syndromes poses significant burden on public healthcare systems in developing countries. With genome-wide diagnostic assays frequently inaccessible, targeted low-cost PCR-based approaches are preferred. However, their reproducibility depends on equally efficient amplification using a number of target and control primers. To address this, the recently described technique called Microdeletion/Microduplication Quantitative Fluorescent PCR (MQF-PCR was shown to reliably detect four human syndromes by quantifying DNA amplification in an internally controlled PCR reaction. Here, we confirm its utility in the detection of eight human microdeletion syndromes, including the more common WAGR, Smith-Magenis, and Potocki-Lupski syndromes with 100% sensitivity and 100% specificity. We present selection, design, and performance evaluation of detection primers using variety of approaches. We conclude that MQF-PCR is an easily adaptable method for detection of human pathological chromosomal aberrations.

  16. A novel and rapid method for obtaining high titre intact prion strains from mammalian brain

    Science.gov (United States)

    Wenborn, Adam; Terry, Cassandra; Gros, Nathalie; Joiner, Susan; D’Castro, Laura; Panico, Silvia; Sells, Jessica; Cronier, Sabrina; Linehan, Jacqueline M.; Brandner, Sebastian; Saibil, Helen R.; Collinge, John; Wadsworth, Jonathan D. F.

    2015-01-01

    Mammalian prions exist as multiple strains which produce characteristic and highly reproducible phenotypes in defined hosts. How this strain diversity is encoded by a protein-only agent remains one of the most interesting and challenging questions in biology with wide relevance to understanding other diseases involving the aggregation or polymerisation of misfolded host proteins. Progress in understanding mammalian prion strains has however been severely limited by the complexity and variability of the methods used for their isolation from infected tissue and no high resolution structures have yet been reported. Using high-throughput cell-based prion bioassay to re-examine prion purification from first principles we now report the isolation of prion strains to exceptional levels of purity from small quantities of infected brain and demonstrate faithful retention of biological and biochemical strain properties. The method’s effectiveness and simplicity should facilitate its wide application and expedite structural studies of prions. PMID:25950908

  17. Comparison of a New and Rapid Method: Brucella Coombs Gel Test With Other Diagnostic Tests.

    Science.gov (United States)

    Kalem, Fatma; Ergün, Ayşe Gül; Durmaz, Süleyman; Doğan, Metin; Ertuğrul, Ömür; Gündem, Seval

    2016-09-01

    The aim of this study was to detect reliability of Brucella Coombs gel test (BCGT) by comparing with with ELISA (IgG + IgM), Standard agglutination test, and Brucella immunocapture agglutination methods in serological diagnosis of brucellosis. Brucella Coombs gel test (BCGT), Brucella ELISA (IgG + IgM), Standard agglutination test, and Brucella immunocapture agglutination tests of 78 patients with presumptive diagnosis of brucellosis which were sent to Microbiology Laboratory of Konya Numune Hospital from various regions of Konya were studied. Of 78 patients with ELISA IgG and IgM, STA, BICA and BCGT; 26, 21, 10, 12 and 12 were positive. When compared with BICA, the sensitivity and specifity of BCGT were 100% and 100%, respectively. According to results BCGT can be used as a diagnostic test in routine laboratories after more comprehensive studies in control groups and patients. © 2016 Wiley Periodicals, Inc.

  18. Rapid and tunable method to temporally control gene editing based on conditional Cas9 stabilization.

    Science.gov (United States)

    Senturk, Serif; Shirole, Nitin H; Nowak, Dawid G; Corbo, Vincenzo; Pal, Debjani; Vaughan, Alexander; Tuveson, David A; Trotman, Lloyd C; Kinney, Justin B; Sordella, Raffaella

    2017-02-22

    The CRISPR/Cas9 system is a powerful tool for studying gene function. Here, we describe a method that allows temporal control of CRISPR/Cas9 activity based on conditional Cas9 destabilization. We demonstrate that fusing an FKBP12-derived destabilizing domain to Cas9 (DD-Cas9) enables conditional Cas9 expression and temporal control of gene editing in the presence of an FKBP12 synthetic ligand. This system can be easily adapted to co-express, from the same promoter, DD-Cas9 with any other gene of interest without co-modulation of the latter. In particular, when co-expressed with inducible Cre-ER T2 , our system enables parallel, independent manipulation of alleles targeted by Cas9 and traditional recombinase with single-cell specificity. We anticipate this platform will be used for the systematic characterization and identification of essential genes, as well as the investigation of the interactions between functional genes.

  19. Rapid Method for Isolation of Desiccation-Tolerant Strains and Xeroprotectants▿

    Science.gov (United States)

    Narváez-Reinaldo, J. J.; Barba, I.; González-López, J.; Tunnacliffe, A.; Manzanera, M.

    2010-01-01

    A novel biotechnological process has been developed for the isolation of desiccation-tolerant microorganisms and their xeroprotectants, i.e., compatible solutes involved in long-term stability of biomolecules in the dry state. Following exposure of soil samples to chloroform, we isolated a collection of desiccation-tolerant microorganisms. This collection was screened for the production of xeroprotectants by a variation of the bacterial milking (osmotic downshock) procedure and by a novel air-drying/rehydration (“dry milking”) incubation method. The resultant solutes were shown to protect both proteins and living cells against desiccation damage, thereby validating them as xeroprotectants. Nuclear magnetic resonance (NMR) analytical studies were performed to identify the xeroprotectants; synthetic mixtures of these compounds were shown to perform similarly to natural isolates in drying experiments with proteins and cells. This new approach has biotechnological and environmental implications for the identification of new xeroprotectants of commercial and therapeutic value. PMID:20562279

  20. SUDOSCAN: A Simple, Rapid, and Objective Method with Potential for Screening for Diabetic Peripheral Neuropathy

    Science.gov (United States)

    Selvarajah, Dinesh; Cash, Tom; Davies, Jennifer; Sankar, Adithya; Rao, Ganesh; Grieg, Marni; Pallai, Shillo; Gandhi, Rajiv; Wilkinson, Iain D.; Tesfaye, Solomon

    2015-01-01

    Clinical methods of detecting diabetic peripheral neuropathy (DPN) are not objective and reproducible. We therefore evaluated if SUDOSCAN, a new method developed to provide a quick, non-invasive and quantitative assessment of sudomotor function can reliably screen for DPN. 70 subjects (45 with type 1 diabetes and 25 healthy volunteers [HV]) underwent detailed assessments including clinical, neurophysiological and 5 standard cardiovascular reflex tests (CARTs). Using the American Academy of Neurology criteria subjects were classified into DPN and No-DPN groups. Based on CARTs subjects were also divided into CAN, subclinical-CAN and no-CAN. Sudomotor function was assessed with measurement of hand and foot Electrochemical Skin Conductance (ESC) and calculation of the CAN risk score. Foot ESC (μS) was significantly lower in subjects with DPN [n = 24; 53.5(25.1)] compared to the No-DPN [77.0(7.9)] and HV [77.1(14.3)] groups (ANCOVA p<0.001). Sensitivity and specificity of foot ESC for classifying DPN were 87.5% and 76.2%, respectively. The area under the ROC curve (AUC) was 0.85. Subjects with CAN had significantly lower foot [55.0(28.2)] and hand [53.5(19.6)] ESC compared to No-CAN [foot ESC, 72.1(12.2); hand ESC 64.9(14.4)] and HV groups (ANCOVA p<0.001 and 0.001, respectively). ROC analysis of CAN risk score to correctly classify CAN revealed a sensitivity of 65.0% and specificity of 80.0%. AUC was 0.75. Both foot and hand ESC demonstrated strong correlation with individual parameters and composite scores of nerve conduction and CAN. SUDOSCAN, a non-invasive and quick test, could be used as an objective screening test for DPN in busy diabetic clinics, insuring adherence to current recommendation of annual assessments for all diabetic patients that remains unfulfilled. PMID:26457582

  1. Rapid and simple methods for quantitative analysis of some antidepressant in pharmaceutical formulations by using first derivative spectrophotometry and HPLC.

    Science.gov (United States)

    Erk, Nevin

    2003-12-01

    Two rapid, simple and accurate first derivative spectrophotometry and HPLC method for the determination of nefazodone hydrochloride and sertraline hydrochloride in pharmaceutical formulations are discussed. The first one is a derivative spectrophotometric procedure and the second one is based on a HPLC method with a UV detector. In the first method, first derivative spectrophotometry, nefazodone hydrochloride or sertraline hydrochloride by measurement of their first derivative signals at 241.8-256.7 nm (peak-to-peak amplitude), or 271.6-275.5 nm (peak-to-peak amplitude), respectively. Calibration graphs were established for 10.0-42.0 microg ml(-1) nefazodone hydrochloride, or 8.0-46.0 microg ml(-1) sertraline hydrochloride. In the other method, HPLC, the UV detection was carried out at 265.0 nm (nefazodone hydrochloride) and 270.0 nm (sertraline hydrochloride). The samples were chromatographed on a Supercosil RP-18 column. The mobile phases were methanol:acetonitrile:phosphate buffer at pH 5.5 (10:50:40 v/v/v) (nefazodone hydrochloride) and methanol:phosphate buffer at pH 4.5 (20:80 v/v) (sertraline hydrochloride). The results obtained from first derivative spectrophotometric method were comparable with those obtained by using HPLC. It was concluded that both the developed methods are equally accurate, sensitive, and precision could be applied directly and easily to the pharmaceutical formulations of nefazodone hydrochloride and sertraline hydrochloride, respectively.

  2. Rapid methods for dioxin and dioxin-like PCBs in food and feedingstuffs. State of the art

    Energy Technology Data Exchange (ETDEWEB)

    Behnisch, P.A. [eurofins-GfA, Muenster (Germany); Hoogenboom, R. [RIKILT-Institute of Food Safety, Wageningen (Netherlands)

    2004-09-15

    The increasing number of local dioxin crises since 2002 becoming more and more apparent due to stricter controls of feed and food in the European Union and the globally increasing number of countries applying similar guidelines make it necessary to establish reliable, time and cost-effective screening methods for the dioxin intake through nutritional pathways. Five years after the last overview presentation about all kinds of different bio-analytical detection methods (BDMs) and the establishment of quality guidelines for screening methods, time has come to include as well the improvements in the chemical methods to speed up the analysis. This review gives an overview about the state-of-the-art improvements and gives a future outlook for both methods, chemical and bio-analytical approach for rapid analyses of dioxins and dioxin-like compounds. Now several new ways of improvement are currently in the pipeline of research and testing, such as PCR, proteomic biomarkers and in case of the clean-up ASE11, PowerPrep and different detection methods as well as different other ways of indicators for dioxins (e.g. correlations to fatty acids).

  3. An Alternative and Rapid Method for the Extraction of Nucleic Acids from Ixodid Ticks by Potassium Acetate Procedure

    Directory of Open Access Journals (Sweden)

    Islay Rodríguez

    2014-08-01

    Full Text Available Four variants of the potassium acetate procedure for DNA extraction from ixodid ticks at different stage of their life cycles were evaluated and compared with phenol-chloroform and ammonium hydroxide methods. The most rapid and most efficient variant was validated in the DNA extraction procedure from the engorged ticks collected from bovine, canine as well as from house ticks for the screening of Borrelia burgdorferi sensu lato, Anaplasma spp. and Babesia spp. The ammonium hydroxide procedure was used for non-engorged ticks. All the variants were efficient and allowed obtaining PCR-quality material according to the specific amplification of 16S rRNA gene fragment of the original tick. DNA extracted from the ticks under the study was tested by multiplex PCR for the screening of tick-borne pathogens. Anaplasma spp. and Babesia spp. amplification products were obtained from 29/48 extracts. Ammonium hydroxide protocol was not efficient for two extracts. Detection of amplification products from the PCR indicated that DNA had been successfully extracted. The potassium acetate procedure could be an alternative, rapid, and reliable method for DNA extraction from the ixodid ticks, mainly for poorly-resourced laboratories.

  4. A rapid method for optimization of the rocket propulsion system for single-stage-to-orbit vehicles

    Science.gov (United States)

    Eldred, C. H.; Gordon, S. V.

    1976-01-01

    A rapid analytical method for the optimization of rocket propulsion systems is presented for a vertical take-off, horizontal landing, single-stage-to-orbit launch vehicle. This method utilizes trade-offs between propulsion characteristics affecting flight performance and engine system mass. The performance results from a point-mass trajectory optimization program are combined with a linearized sizing program to establish vehicle sizing trends caused by propulsion system variations. The linearized sizing technique was developed for the class of vehicle systems studied herein. The specific examples treated are the optimization of nozzle expansion ratio and lift-off thrust-to-weight ratio to achieve either minimum gross mass or minimum dry mass. Assumed propulsion system characteristics are high chamber pressure, liquid oxygen and liquid hydrogen propellants, conventional bell nozzles, and the same fixed nozzle expansion ratio for all engines on a vehicle.

  5. A comparative study of methods for automatic detection of rapid eye movement abnormal muscular activity in narcolepsy

    DEFF Research Database (Denmark)

    Olesen, Alexander Neergaard; Cesari, Matteo; Christensen, Julie Anja Engelhard

    2018-01-01

    Objective: To evaluate rapid eye movement (REM) muscular activity in narcolepsy by applying five algorithms to electromyogram (EMG) recordings, and to investigate its value for narcolepsy diagnosis. Patients/methods: A modified version of phasic EMG metric (mPEM), muscle activity index (MAI), REM...... atonia index (RAI), supra-threshold REM EMG activit ymetric (STREAM), and Frandsen method (FR) were calculated from polysomnography recordings of 20 healthy controls, 18 clinic controls (subjects suspected with narcolepsy but finally diagnosed without any sleep abnormality), 16 narcolepsy type 1 without...... REM sleep behavior disorder (RBD), 9 narcolepsy type 1 with RBD, and 18 narcolepsy type 2. Diagnostic value of metrics in differentiating between groups was quantified by area under the receiver operating characteristic curve (AUC). Correlations among the metrics and cerebrospinal fluid hypocretin-1...

  6. [A rapid method for continuous flow measurement of cholesterol contained in high density lipoproteins (HDL) (author's transl)].

    Science.gov (United States)

    Ponsot, P; Yvert, J P; Chevrier, M; Bon, R

    1981-01-01

    The authors utilized a reagent containing concanavalin A, a vegetal lecithin, to selectively precipitate lipoproteins containing apoprotein B, a component of VLDL, LDL, and Lp (a) which are well known for their atherogenic risk. During this precipitation "true" high density lipoproteins remain in solution. HDL cholesterol determination which constitutes an indirect indication of HDL activity or concentrations is performed by an enzymatic method using an automated continuous flow technique carried out on an Auto Analyzer II (Technicon Corp.). This rapid, easy determination obtains results comparable to other methods, particularly those chosen by the Société Française de Biologie Clinique (French Society of Clinical Biology). This technique should permit all laboratories to confirm an atherogenic index.

  7. Rapid and Checkable Electrical Post-Treatment Method for Organic Photovoltaic Devices

    Science.gov (United States)

    Park, Sangheon; Seo, Yu-Seong; Shin, Won Suk; Moon, Sang-Jin; Hwang, Jungseek

    2016-01-01

    Post-treatment processes improve the performance of organic photovoltaic devices by changing the microscopic morphology and configuration of the vertical phase separation in the active layer. Thermal annealing and solvent vapor (or chemical) treatment processes have been extensively used to improve the performance of bulk-heterojunction (BHJ) organic photovoltaic (OPV) devices. In this work we introduce a new post-treatment process which we apply only electrical voltage to the BHJ-OPV devices. We used the commercially available P3HT [Poly(3-hexylthiophene)] and PC61BM (Phenyl-C61-Butyric acid Methyl ester) photovoltaic materials as donor and acceptor, respectively. We monitored the voltage and current applied to the device to check for when the post-treatment process had been completed. This electrical treatment process is simpler and faster than other post-treatment methods, and the performance of the electrically treated solar cell is comparable to that of a reference (thermally annealed) device. Our results indicate that the proposed treatment process can be used efficiently to fabricate high-performance BHJ-OPV devices. PMID:26932767

  8. A rapid method for the sampling of atmospheric water vapour for isotopic analysis.

    Science.gov (United States)

    Peters, Leon I; Yakir, Dan

    2010-01-01

    Analysis of the stable isotopic composition of atmospheric moisture is widely applied in the environmental sciences. Traditional methods for obtaining isotopic compositional data from ambient moisture have required complicated sampling procedures, expensive and sophisticated distillation lines, hazardous consumables, and lengthy treatments prior to analysis. Newer laser-based techniques are expensive and usually not suitable for large-scale field campaigns, especially in cases where access to mains power is not feasible or high spatial coverage is required. Here we outline the construction and usage of a novel vapour-sampling system based on a battery-operated Stirling cycle cooler, which is simple to operate, does not require any consumables, or post-collection distillation, and is light-weight and highly portable. We demonstrate the ability of this system to reproduce delta(18)O isotopic compositions of ambient water vapour, with samples taken simultaneously by a traditional cryogenic collection technique. Samples were collected over 1 h directly into autosampler vials and were analysed by mass spectrometry after pyrolysis of 1 microL aliquots to CO. This yielded an average error of distillation lines, slurry maintenance or mains power is not feasible. Copyright 2009 John Wiley & Sons, Ltd.

  9. A simple and rapid method for standard preparation of gas phase extract of cigarette smoke.

    Directory of Open Access Journals (Sweden)

    Tsunehito Higashi

    Full Text Available Cigarette smoke consists of tar and gas phase: the latter is toxicologically important because it can pass through lung alveolar epithelium to enter the circulation. Here we attempt to establish a standard method for preparation of gas phase extract of cigarette smoke (CSE. CSE was prepared by continuously sucking cigarette smoke through a Cambridge filter to remove tar, followed by bubbling it into phosphate-buffered saline (PBS. An increase in dry weight of the filter was defined as tar weight. Characteristically, concentrations of CSEs were represented as virtual tar concentrations, assuming that tar on the filter was dissolved in PBS. CSEs prepared from smaller numbers of cigarettes (original tar concentrations ≤ 15 mg/ml showed similar concentration-response curves for cytotoxicity versus virtual tar concentrations, but with CSEs from larger numbers (tar ≥ 20 mg/ml, the curves were shifted rightward. Accordingly, the cytotoxic activity was detected in PBS of the second reservoir downstream of the first one with larger numbers of cigarettes. CSEs prepared from various cigarette brands showed comparable concentration-response curves for cytotoxicity. Two types of CSEs prepared by continuous and puff smoking protocols were similar regarding concentration-response curves for cytotoxicity, pharmacology of their cytotoxicity, and concentrations of cytotoxic compounds. These data show that concentrations of CSEs expressed by virtual tar concentrations can be a reference value to normalize their cytotoxicity, irrespective of numbers of combusted cigarettes, cigarette brands and smoking protocols, if original tar concentrations are ≤15 mg/ml.

  10. A new real-time PCR method for rapid and specific detection of ling (Molva molva).

    Science.gov (United States)

    Taboada, Ledicia; Sánchez, Ana; Sotelo, Carmen G

    2017-08-01

    Seafood fraud - often involving substitution of one species by another - has attracted much attention as it is prevalent worldwide. Whilst DNA analysis has helped to combat this type of fraud some of the methods currently in use are time-consuming and require sophisticated equipment or highly-trained personnel. This work describes the development of a new, real-time PCR TaqMan assay for the detection of ling (Molva molva) in seafood products. For this purpose, specific primers and a minor groove binding (MGB) TaqMan probe were designed to amplify the 81bp region on the cyt b gene. Efficiency, specificity and cross-reactivity assays showed statistically significant differences between the average Ct value obtained for Molva molva DNA (19.45±0.65) and the average Ct for non-target species DNA (38.3±2.8), even with closely related species such as Molva dypterygia (34.9±0.09). The proposed methodology has been validated with 31 commercial samples. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. An easy, rapid method to isolate RPE cell protein from the mouse eye.

    Science.gov (United States)

    Wei, Hong; Xun, Zixian; Granado, Herta; Wu, Angela; Handa, James T

    2016-04-01

    The retinal pigment epithelium (RPE) is essential for maintaining the health of the neural retina. RPE cell dysfunction plays a critical role in many common blinding diseases including age-related macular degeneration (AMD), diabetic retinopathy, retinal dystrophies. Mouse models of ocular disease are commonly used to study these blinding diseases. Since isolating the RPE from the choroid has been challenging, most techniques separate the RPE from the retina, but not the choroid. As a result, the protein signature actually represents a heterogeneous population of cells that may not accurately represent the RPE response. Herein, we describe a method for separating proteins from the RPE that is free from retinal and choroidal contamination. After removing the anterior segment and retina from enucleated mouse eyes, protein from the RPE was extracted separately from the choroid by incubating the posterior eyecup with a protein lysis buffer for 10 min. Western blot analysis identified RPE65, an RPE specific protein in the RPE lysates, but not in choroidal lysates. The RPE lysates were devoid of rhodopsin and collagen VI, which are abundant in the retina and choroid, respectively. This technique will be very helpful for measuring the protein signal from the RPE without retinal or choroidal contamination. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Advanced Mass Spectrometric Methods for the Rapid and Quantitative Characterization of Proteomes

    Directory of Open Access Journals (Sweden)

    Richard D. Smith

    2006-04-01

    Full Text Available Progress is reviewed towards the development of a global strategy that aims to extend the sensitivity, dynamic range, comprehensiveness and throughput of proteomic measurements based upon the use of high performance separations and mass spectrometry. The approach uses high accuracy mass measurements from Fourier transform ion cyclotron resonance mass spectrometry (FTICR to validate peptide ‘accurate mass tags’ (AMTs produced by global protein enzymatic digestions for a specific organism, tissue or cell type from ‘potential mass tags’ tentatively identified using conventional tandem mass spectrometry (MS/MS. This provides the basis for subsequent measurements without the need for MS/ MS. High resolution capillary liquid chromatography separations combined with high sensitivity, and high resolution accurate FTICR measurements are shown to be capable of characterizing peptide mixtures of more than 105 components. The strategy has been initially demonstrated using the microorganisms Saccharomyces cerevisiae and Deinococcus radiodurans. Advantages of the approach include the high confidence of protein identification, its broad proteome coverage, high sensitivity, and the capability for stableisotope labeling methods for precise relative protein abundance measurements.

  13. A Simple and Rapid Method for Standard Preparation of Gas Phase Extract of Cigarette Smoke

    Science.gov (United States)

    Higashi, Tsunehito; Mai, Yosuke; Noya, Yoichi; Horinouchi, Takahiro; Terada, Koji; Hoshi, Akimasa; Nepal, Prabha; Harada, Takuya; Horiguchi, Mika; Hatate, Chizuru; Kuge, Yuji; Miwa, Soichi

    2014-01-01

    Cigarette smoke consists of tar and gas phase: the latter is toxicologically important because it can pass through lung alveolar epithelium to enter the circulation. Here we attempt to establish a standard method for preparation of gas phase extract of cigarette smoke (CSE). CSE was prepared by continuously sucking cigarette smoke through a Cambridge filter to remove tar, followed by bubbling it into phosphate-buffered saline (PBS). An increase in dry weight of the filter was defined as tar weight. Characteristically, concentrations of CSEs were represented as virtual tar concentrations, assuming that tar on the filter was dissolved in PBS. CSEs prepared from smaller numbers of cigarettes (original tar concentrations ≤15 mg/ml) showed similar concentration-response curves for cytotoxicity versus virtual tar concentrations, but with CSEs from larger numbers (tar ≥20 mg/ml), the curves were shifted rightward. Accordingly, the cytotoxic activity was detected in PBS of the second reservoir downstream of the first one with larger numbers of cigarettes. CSEs prepared from various cigarette brands showed comparable concentration-response curves for cytotoxicity. Two types of CSEs prepared by continuous and puff smoking protocols were similar regarding concentration-response curves for cytotoxicity, pharmacology of their cytotoxicity, and concentrations of cytotoxic compounds. These data show that concentrations of CSEs expressed by virtual tar concentrations can be a reference value to normalize their cytotoxicity, irrespective of numbers of combusted cigarettes, cigarette brands and smoking protocols, if original tar concentrations are ≤15 mg/ml. PMID:25229830

  14. A simple and rapid chromatographic method to determine unauthorized basic colorants (rhodamine B, auramine O, and pararosaniline) in processed foods

    Science.gov (United States)

    Tatebe, Chiye; Zhong, Xining; Ohtsuki, Takashi; Kubota, Hiroki; Sato, Kyoko; Akiyama, Hiroshi

    2014-01-01

    A simple and rapid high-performance liquid chromatography (HPLC) method to determine basic colorants such as pararosaniline (PA), auramine O (AO), and rhodamine B (RB) in various processed foods was developed. Linearity of the calibration curves ranged from 0.05 to 50 μg/mL for PA and 0.05–100 μg/mL for AO and RB. The detection and quantification limits (LOD and LOQ) of the basic colorants, which were evaluated as signal-to-noise ratios of 3 for LOD and 10 for LOQ, ranged from 0.0125 to 0.05 and 0.025 to 0.125 μg/g, respectively. The recoveries and relative standard deviations of three basic colorants in six processed foods, namely, chili sauce, curry paste, gochujang (hot pepper paste), tandoori chicken (roasted chicken prepared with yogurt and spices), powder soup, and shrimp powder ranged from 70.2% to 102.8% and 0.8% to 8.0%, respectively. The intraday precision of the recovery test ranged from 1.7% to 4.5%, whereas the interday precision ranged from 3.7% to 7.7%. The reported method has been successfully applied to basic colorant determination in various processed foods such as fat-based food matrices (curry paste and tandoori chicken), chili products (gochujang and chili sauce), and protein-based products (shrimp powder and powder soup). Thin layer chromatography and liquid chromatography/mass spectrometry methods for the determination of basic colorants in processed foods were also developed for rapid analysis and identification, respectively. These methods are very useful for monitoring unauthorized basic colorants in inspection centers or quarantine laboratories in many countries. PMID:25473512

  15. Enumeration of coliforms and Escherichia coli in frozen black tiger shrimp Penaeus monodon by conventional and rapid methods.

    Science.gov (United States)

    Suwansonthichai, Sasithorn; Rengpipat, Sirirat

    2003-03-15

    Conventional (most probable number, MPN) and rapid methods-including Chromocult coliform agar (CCA), Fluorocult(R) LMX broth (LMX), and Petrifilm Escherichia coli count plates (PEC) for enumeration of coliforms and E. coli in frozen black tiger shrimp from Thailand were compared in order to assess the possibility of using one of the rapid methods for routine analysis. Enumeration of coliforms and E. coli from 18 samples of regular frozen black tiger shrimp and 156 samples of frozen black tiger shrimp experimentally contaminated with coliforms or E. coli at concentrations of approximately 10, approximately 10(2), and approximately 10(3) CFU g(-1) revealed that at the level of approximately 10 CFU g(-1), coliform numbers ranked as LMX>CCA>MPN=PEC and E. coli as MPN=LMX=PEC=CCA. At the level of approximately 10(2) CFU g(-1), coliform numbers ranked as LMX>MPN=PEC=CCA and E. coli as MPN=LMX>PEC=CCA. At the level of 10(3) CFU g(-1), coliforms ranked as LMX>MPN=CCA>PEC and E. coli as MPN>LMX>CCA>PEC. Agreements with the conventional MPN method for coliforms were LMX 108%, PEC 87.2%, and CCA 91.2% and agreements for E. coli were LMX 101%, PEC 95.7%, and CCA 96.3%. Sensitivities (%) ranked LMX>MPN>CCA=PEC for coliforms and E. coli, whereas equal specificities (100%) of all methods for coliforms and E. coli were demonstrated. Rankings for the other parameters compared were: convenience, PEC>CCA=LMX>MPN; time to detection, MPN>LMX=PEC=CCA; expense, MPN=PEC>CCA>LMX; labor, MPN>LMX=CCA>PEC; accuracy for coliforms, PEC>CCA>MPN>LMX; and accuracy for E. coli, PEC=CCA>LMX>MPN.

  16. Interlaboratory validation of a real-time PCR 24-hour rapid method for detection of Salmonella in foods.

    Science.gov (United States)

    Cheng, Chorng-Ming; Van Khanh, T; Lin, Wen; Ruby, Richard M

    2009-05-01

    The efficacy of a 24-h Salmonella real-time, or quantitative, PCR (qPCR) detection method was assessed through a collaborative effort involving eight Federal and state laboratories. Eleven foods including mashed potatoes, soft cheese, chili powder, chocolate, eggs, sprouts, apple juice, fish, shrimp, ground beef, and ground chicken were tested. For each food, seven blind samples were distributed to each participant for testing. These included six samples equivalently inoculated with 1 to 5 CFU/25 g of various serotypes of Salmonella (Gaminara, Weltevreden, Heidelberg, Senftenberg, Enteritidis, Newport, Typhimurium, and Kentucky for each food) and 10 to 50 CFU/25 g of the competitor Enterobacter cloacae. The seventh sample was inoculated with 10 to 50 CFU/25 g of the competitor, E. cloacae, only. These samples were tested for Salmonella by using four methods in parallel: (i) 24-h qPCR method detecting Salmonella from modified buffered peptone water enrichment medium; (ii) 48-h qPCR method detecting Salmonella from a secondary selective enrichment broth; (iii) modified Bacteriological Analytical Manual method; and (iv) VIDAS, an immunoassay system. The results of the statistical analysis showed there was no significant (P > or = 0.05) difference between either of the qPCR methods and the modified Bacteriological Analytical Manual method for 10 of 11 foods. For the one exception, sprouts, detection by qPCR required 48 h. Both qPCR methods showed a detection limit of 0.08 to 0.2 CFU/g. These results provide a solid basis for using this 24-h qPCR rapid screening method to detect Salmonella in foods.

  17. Electromagnetic methods for measuring materials properties of cylindrical rods and array probes for rapid flaw inspection

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Haiyan [Iowa State Univ., Ames, IA (United States)

    2005-01-01

    The case-hardening process modifies the near-surface permeability and conductivity of steel, as can be observed through changes in alternating current potential drop (ACPD) along a rod. In order to evaluate case depth of case hardened steel rods, analytical expressions are derived for the alternating current potential drop on the surface of a homogeneous rod, a two-layered and a three-layered rod. The case-hardened rod is first modeled by a two-layer rod that has a homogeneous substrate with a single, uniformly thick, homogeneous surface layer, in which the conductivity and permeability values differ from those in the substrate. By fitting model results to multi-frequency ACPD experimental data, estimates of conductivity, permeability and case depth are found. Although the estimated case depth by the two-layer model is in reasonable agreement with the effective case depth from the hardness profile, it is consistently higher than the effective case depth. This led to the development of the three-layer model. It is anticipated that the new three-layered model will improve the results and thus makes the ACPD method a novel technique in nondestructive measurement of case depth. Another way to evaluate case depth of a case hardened steel rod is to use induction coils. Integral form solutions for an infinite rod encircled by a coaxial coil are well known, but for a finite length conductor, additional boundary conditions must be satisfied at the ends. In this work, calculations of eddy currents are performed for a two-layer conducting rod of finite length excited by a coaxial circular coil carrying an alternating current. The solution is found using the truncated region eigenfunction expansion (TREE) method. By truncating the solution region to a finite length in the axial direction, the magnetic vector potential can be expressed as a series expansion of orthogonal eigenfunctions instead of as a Fourier integral. Closed-form expressions are derived for the electromagnetic

  18. A rapid method for the determination of brominated flame retardant concentrations in plastics and textiles entering the waste stream.

    Science.gov (United States)

    Abdallah, Mohamed Abou-Elwafa; Drage, Daniel S; Sharkey, Martin; Berresheim, Harald; Harrad, Stuart

    2017-10-01

    Due to new European legislation, products going to waste are subject to 'low persistent organic pollutant concentration limits'. Concentrations of restricted brominated flame retardants in waste products must be determined. A rapid extraction and clean-up method was developed for determination of brominated flame retardants in various plastics and textiles. The optimised method used vortexing and ultrasonication in dichloromethane followed by sulfuric acid clean-up to determine target compounds. Poly-brominated diphenyl ethers were determined by gas chromatography with mass spectrometry and hexabromocyclododecane by liquid chromatography with tandem mass spectrometry. Good recoveries of target analytes were obtained after three extraction cycles. The method was validated using poly-propylene and poly-ethylene certified reference materials as well as previously characterised textiles, expanded and extruded poly-styrene samples. Measured concentrations of target compounds showed good agreement with the certified values indicating good accuracy and precision. Clean extracts provided low noise levels resulting in low limits of quantification (0.8-1.5 ng/g for poly-brominated diphenyl ethers and 0.3 ng/g for α-, β- and γ-hexabromocyclododecane). The developed method was applied successfully to real consumer products entering the waste stream and it provided various advantages over traditional methods, including reduced analysis time, solvent consumption, minimal sample contamination and high sample throughput, which is crucial to comply with the implemented legislation. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Rapid Automatic Lighting Control of a Mixed Light Source for Image Acquisition using Derivative Optimum Search Methods

    Directory of Open Access Journals (Sweden)

    Kim HyungTae

    2015-01-01

    Full Text Available Automatic lighting (auto-lighting is a function that maximizes the image quality of a vision inspection system by adjusting the light intensity and color.In most inspection systems, a single color light source is used, and an equal step search is employed to determine the maximum image quality. However, when a mixed light source is used, the number of iterations becomes large, and therefore, a rapid search method must be applied to reduce their number. Derivative optimum search methods follow the tangential direction of a function and are usually faster than other methods. In this study, multi-dimensional forms of derivative optimum search methods are applied to obtain the maximum image quality considering a mixed-light source. The auto-lighting algorithms were derived from the steepest descent and conjugate gradient methods, which have N-size inputs of driving voltage and one output of image quality. Experiments in which the proposed algorithm was applied to semiconductor patterns showed that a reduced number of iterations is required to determine the locally maximized image quality.

  20. Comparison of various molecular methods for rapid differentiation of intestinal bifidobacteria at the species, subspecies and strain level.

    Science.gov (United States)

    Jarocki, Piotr; Podleśny, Marcin; Komoń-Janczara, Elwira; Kucharska, Jagoda; Glibowska, Agnieszka; Targoński, Zdzisław

    2016-07-22

    Members of the genus Bifidobacterium are anaerobic Gram-positive Actinobacteria, which are natural inhabitants of human and animal gastrointestinal tract. Certain bifidobacteria are frequently used as food additives and probiotic pharmaceuticals, because of their various health-promoting properties. Due to the enormous demand on probiotic bacteria, manufacture of high-quality products containing living microorganisms requires rapid and accurate identification of specific bacteria. Additionally, isolation of new industrial bacteria from various environments may lead to multiple isolations of the same strain, therefore, it is important to apply rapid, low-cost and effective procedures differentiating bifidobacteria at the intra-species level. The identification of new isolates using microbiological and biochemical methods is difficult, but the accurate characterization of isolated strains may be achieved using a polyphasic approach that includes classical phenotypic methods and molecular procedures. However, some of these procedures are time-consuming and cumbersome, particularly when a large group of new isolates is typed, while some other approaches may have too low discriminatory power to distinguish closely related isolates obtained from similar sources. This work presents the evaluation of the discriminatory power of four molecular methods (ARDRA, RAPD-PCR, rep-PCR and SDS-PAGE fingerprinting) that are extensively used for fast differentiation of bifidobacteria up to the strain level. Our experiments included 17 reference strains and showed that in comparison to ARDRA, genotypic fingerprinting procedures (RAPD and rep-PCR) seemed to be less reproducible, however, they allowed to differentiate the tested microorganisms even at the intra-species level. In general, RAPD and rep-PCR have similar discriminatory power, though, in some instances more than one oligonucleotide needs to be used in random amplified polymorphic DNA analysis. Moreover, the results also

  1. Comparison of the Immunomagnetic Separation/Adenosine Triphosphate Rapid Method and the Modified mTEC Membrane-Filtration Method for Enumeration of Escherichia coli

    Science.gov (United States)

    Brady, Amie M.G.; Bushon, Rebecca N.; Bertke, Erin E.

    2009-01-01

    Water quality at beaches is monitored for fecal indicator bacteria by traditional, culture-based methods that can take 18 to 24 hours to obtain results. A rapid detection method that provides estimated concentrations of fecal indicator bacteria within 1 hour from the start of sample processing would allow beach managers to post advisories or close the beach when the conditions are actually considered unsafe instead of a day later, when conditions may have changed. A rapid method that couples immunomagnetic separation with adenosine triphosphate detection (IMS/ATP rapid method) was evaluated through monitoring of Escherichia coli (E. coli) at three Lake Erie beaches in Ohio (Edgewater and Villa Angela in Cleveland and Huntington in Bay Village). Beach water samples were collected between 4 and 5 days per week during the recreational seasons (May through September) of 2006 and 2007. Composite samples were created in the lab from two point samples collected at each beach and were shown to be comparable substitutes for analysis of two individual samples. E. coli concentrations in composite samples, as determined by the culture-based method, ranged from 4 to 24,000 colony-forming units per 100 milliliters during this study across all beaches. Turbidity also was measured for each sample and ranged from 0.8 to 260 neophelometric turbidity ratio units. Environmental variables were noted at the time of sampling, including number of birds at the beach and wave height. Rainfall amounts were measured at National Weather Service stations at local airports. Turbidity, rainfall, and wave height were significantly related to the culture-based method results each year and for both years combined at each beach. The number of birds at the beach was significantly related to the culture-based method results only at Edgewater during 2006 and during both years combined. Results of the IMS/ATP method were compared to results of the culture-based method for samples by year for each beach

  2. Method to Rapidly Collect Thousands of Velocity Observations to Validate Million-Element 2D Hydrodynamic Models

    Science.gov (United States)

    Barker, J. R.; Pasternack, G. B.; Bratovich, P.; Massa, D.; Reedy, G.; Johnson, T.

    2010-12-01

    Two-dimensional (depth-averaged) hydrodynamic models have existed for decades and are used to study a variety of hydrogeomorphic processes as well as to design river rehabilitation projects. Rapid computer and coding advances are revolutionizing the size and detail of 2D models. Meanwhile, advances in topo mapping and environmental informatics are providing the data inputs to drive large, detailed simulations. Million-element computational meshes are in hand. With simulations of this size and detail, the primary challenge has shifted to finding rapid and inexpensive means for testing model predictions against observations. Standard methods for collecting velocity data include boat-mounted ADCP and point-based sensors on boats or wading rods. These methods are labor intensive and often limited to a narrow flow range. Also, they generate small datasets at a few cross-sections, which is inadequate to characterize the statistical structure of the relation between predictions and observations. Drawing on the long-standing oceanographic method of using drogues to track water currents, previous studies have demonstrated the potential of small dGPS units to obtain surface velocity in rivers. However, dGPS is too inaccurate to test 2D models. Also, there is financial risk in losing drogues in rough currents. In this study, an RTK GPS unit was mounted onto a manned whitewater kayak. The boater positioned himself into the current and used floating debris to maintain a speed and heading consistent with the ambient surface flow field. RTK GPS measurements were taken ever 5 sec. From these positions, a 2D velocity vector was obtained. The method was tested over ~20 km of the lower Yuba River in California in flows ranging from 500-5000 cfs, yielding 5816 observations. To compare velocity magnitude against the 2D model-predicted depth-averaged value, kayak-based surface values were scaled down by an optimized constant (0.72), which had no negative effect on regression analysis

  3. Validation of a rapid lateral flow method for the detection of cows' milk in water buffalo, sheep or goat milk.

    Science.gov (United States)

    Galan-Malo, Patricia; Mendiara, Isabel; Razquin, Pedro; Mata, Luis

    2018-01-22

    For many years, the adulteration of milk from sheep, goats or water buffalos with cows' milk has been a widespread practice due to the higher cost of milk from those other species. Because of this, great concern has been shown by many Protected Designation of Origin councils that have to assure the quality and genuineness of the cheese produced by their associates. Therefore, the whole production chain needs analytical tools that allow the control of potential adulteration. Rapid methods to be used in the field are scarce and have not been validated according to international guidelines. The aim of this work has been to validate a rapid test based on lateral flow immunochromatography to detect cows' milk in milk from other species, including buffalo's milk, according to AOAC guidelines. No false-positive result was found after analysing 146 known negative samples from individual animals. The lowest level of adulteration with a Probability of Detection (POD) of 1.00 (confidence interval between 0.94 and 1.00) was found at 0.5% of cows' milk. This level is below the current EU allowed level of cows' milk, set at 1%. Variations in the time of assay, volume of the analysis buffer and different batches of the test were evaluated to detect any effect on the false-positive rate or on the limit of detection of the test. The effects of compositional factors (such as high level of fat, protein and somatic cell counts) were also evaluated. The new rapid test to detect cows' milk in milk from other species is shown to be an adequate tool to control milk quality in routine analysis. This kind of test is very easy to use and it can be performed by untrained staff during milk collection at the farm or upon arrival at dairies.

  4. Rapid validated HPTLC method for estimation of piperine and piperlongumine in root of Piper longum extract and its commercial formulation

    Directory of Open Access Journals (Sweden)

    Anagha A. Rajopadhye

    2012-12-01

    Full Text Available Piperine and piperlongumine, alkaloids having diverse biological activities, commonly occur in roots of Piper longum L., Piperaceae, which have high commercial, economical and medicinal value. In present study, rapid, validated HPTLC method has been established for the determination of piperine and piperlongumine in methanolic root extract and its commercial formulation 'Mahasudarshan churna®' using ICH guidelines. The use of Accelerated Solvent Extraction (ASE as an alternative to conventional techniques has been explored. The methanol extracts of root, its formulation and both standard solutions were applied on silica gel F254 HPTLC plates. The plates were developed in Twin chamber using mobile phase toluene: ethyl acetate (6:4, v/v and scanned at 342 and 325 nm (λmax of piperine and piperlongumine, respectively using Camag TLC scanner 3 with CATS 4 software. A linear relationship was obtained between response (peak area and amount of piperine and piperlongumine in the range of 20-100 and 30-150 ng/spot, respectively; the correlation coefficient was 0.9957 and 0.9941 respectively. Sharp, symmetrical and well resolved peaks of piperine and piperlongumine spots resolved at Rf 0.51 and 0.74, respectively from other components of the sample extracts. The HPTLC method showed good linearity, recovery and high precision of both markers. Extraction of plant using ASE and rapid HPTLC method provides a new and powerful approach to estimate piperine and piperlongumine as phytomarkers in the extract as well as its commercial formulations for routine quality control.

  5. Rapid validated HPTLC method for estimation of piperine and piperlongumine in root of Piper longum extract and its commercial formulation

    Directory of Open Access Journals (Sweden)

    Anagha A. Rajopadhye

    2012-09-01

    Full Text Available Piperine and piperlongumine, alkaloids having diverse biological activities, commonly occur in roots of Piper longum L., Piperaceae, which have high commercial, economical and medicinal value. In present study, rapid, validated HPTLC method has been established for the determination of piperine and piperlongumine in methanolic root extract and its commercial formulation 'Mahasudarshan churna®' using ICH guidelines. The use of Accelerated Solvent Extraction (ASE as an alternative to conventional techniques has been explored. The methanol extracts of root, its formulation and both standard solutions were applied on silica gel F254 HPTLC plates. The plates were developed in Twin chamber using mobile phase toluene: ethyl acetate (6:4, v/v and scanned at 342 and 325 nm (λmax of piperine and piperlongumine, respectively using Camag TLC scanner 3 with CATS 4 software. A linear relationship was obtained between response (peak area and amount of piperine and piperlongumine in the range of 20-100 and 30-150 ng/spot, respectively; the correlation coefficient was 0.9957 and 0.9941 respectively. Sharp, symmetrical and well resolved peaks of piperine and piperlongumine spots resolved at Rf 0.51 and 0.74, respectively from other components of the sample extracts. The HPTLC method showed good linearity, recovery and high precision of both markers. Extraction of plant using ASE and rapid HPTLC method provides a new and powerful approach to estimate piperine and piperlongumine as phytomarkers in the extract as well as its commercial formulations for routine quality control.

  6. Cochrane Rapid Reviews Methods Group to play a leading role in guiding the production of informed high-quality, timely research evidence syntheses

    Directory of Open Access Journals (Sweden)

    Chantelle Garritty

    2016-10-01

    Full Text Available Abstract Background Policymakers and healthcare stakeholders are increasingly seeking evidence to inform the policymaking process, and often use existing or commissioned systematic reviews to inform decisions. However, the methodologies that make systematic reviews authoritative take time, typically 1 to 2 years to complete. Outside the traditional SR timeline, “rapid reviews” have emerged as an efficient tool to get evidence to decision-makers more quickly. However, the use of rapid reviews does present challenges. To date, there has been limited published empirical information about this approach to compiling evidence. Thus, it remains a poorly understood and ill-defined set of diverse methodologies with various labels. In recent years, the need to further explore rapid review methods, characteristics, and their use has been recognized by a growing network of healthcare researchers, policymakers, and organizations, several with ties to Cochrane, which is recognized as representing an international gold standard for high-quality, systematic reviews. Purpose In this commentary, we introduce the newly established Cochrane Rapid Reviews Methods Group developed to play a leading role in guiding the production of rapid reviews given they are increasingly employed as a research synthesis tool to support timely evidence-informed decision-making. We discuss how the group was formed and outline the group’s structure and remit. We also discuss the need to establish a more robust evidence base for rapid reviews in the published literature, and the importance of promoting registration of rapid review protocols in an effort to promote efficiency and transparency in research. Conclusion As with standard systematic reviews, the core principles of evidence-based synthesis should apply to rapid reviews in order to minimize bias to the extent possible. The Cochrane Rapid Reviews Methods Group will serve to establish a network of rapid review stakeholders

  7. Testing survey-based methods for rapid monitoring of child mortality, with implications for summary birth history data.

    Science.gov (United States)

    Brady, Eoghan; Hill, Kenneth

    2017-01-01

    Under-five mortality estimates are increasingly used in low and middle income countries to target interventions and measure performance against global development goals. Two new methods to rapidly estimate under-5 mortality based on Summary Birth Histories (SBH) were described in a previous paper and tested with data available. This analysis tests the methods using data appropriate to each method from 5 countries that lack vital registration systems. SBH data are collected across many countries through censuses and surveys, and indirect methods often rely upon their quality to estimate mortality rates. The Birth History Imputation method imputes data from a recent Full Birth History (FBH) onto the birth, death and age distribution of the SBH to produce estimates based on the resulting distribution of child mortality. DHS FBHs and MICS SBHs are used for all five countries. In the implementation, 43 of 70 estimates are within 20% of validation estimates (61%). Mean Absolute Relative Error is 17.7.%. 1 of 7 countries produces acceptable estimates. The Cohort Change method considers the differences in births and deaths between repeated Summary Birth Histories at 1 or 2-year intervals to estimate the mortality rate in that period. SBHs are taken from Brazil's PNAD Surveys 2004-2011 and validated against IGME estimates. 2 of 10 estimates are within 10% of validation estimates. Mean absolute relative error is greater than 100%. Appropriate testing of these new methods demonstrates that they do not produce sufficiently good estimates based on the data available. We conclude this is due to the poor quality of most SBH data included in the study. This has wider implications for the next round of censuses and future household surveys across many low- and middle- income countries.

  8. Comparison of immunomagnetic separation/adenosine triphosphate rapid method to traditional culture-based method for E. coli and enterococci enumeration in wastewater

    Science.gov (United States)

    Bushon, R.N.; Likirdopulos, C.A.; Brady, A.M.G.

    2009-01-01

    Untreated wastewater samples from California, North Carolina, and Ohio were analyzed by the immunomagnetic separation/adenosine triphosphate (IMS/ATP) method and the traditional culture-based method for E. coli and enterococci concentrations. The IMS/ATP method concentrates target bacteria by immunomagnetic separation and then quantifies captured bacteria by measuring bioluminescence induced by release of ATP from the bacterial cells. Results from this method are available within 1 h from the start of sample processing. Significant linear correlations were found between the IMS/ATP results and results from traditional culture-based methods for E. coli and enterococci enumeration for one location in California, two locations in North Carolina, and one location in Ohio (r??values ranged from 0.87 to 0.97). No significant linear relation was found for a second location in California that treats a complex mixture of residential and industrial wastewater. With the exception of one location, IMS/ATP showed promise as a rapid method for the quantification of faecal-indicator organisms in wastewater.

  9. Rapid detection of coliforms in drinking water of Arak city using multiplex PCR method in comparison with the standard method of culture (Most Probably Number)

    Science.gov (United States)

    Fatemeh, Dehghan; Reza, Zolfaghari Mohammad; Mohammad, Arjomandzadegan; Salomeh, Kalantari; Reza, Ahmari Gholam; Hossein, Sarmadian; Maryam, Sadrnia; Azam, Ahmadi; Mana, Shojapoor; Negin, Najarian; Reza, Kasravi Alii; Saeed, Falahat

    2014-01-01

    Objective To analyse molecular detection of coliforms and shorten the time of PCR. Methods Rapid detection of coliforms by amplification of lacZ and uidA genes in a multiplex PCR reaction was designed and performed in comparison with most probably number (MPN) method for 16 artificial and 101 field samples. The molecular method was also conducted on isolated coliforms from positive MPN samples; standard sample for verification of microbial method certificated reference material; isolated strains from certificated reference material and standard bacteria. The PCR and electrophoresis parameters were changed for reducing the operation time. Results Results of PCR for lacZ and uidA genes were similar in all of standard, operational and artificial samples and showed the 876 bp and 147 bp bands of lacZ and uidA genes by multiplex PCR. PCR results were confirmed by MPN culture method by sensitivity 86% (95% CI: 0.71-0.93). Also the total execution time, with a successful change of factors, was reduced to less than two and a half hour. Conclusions Multiplex PCR method with shortened operation time was used for the simultaneous detection of total coliforms and Escherichia coli in distribution system of Arak city. It's recommended to be used at least as an initial screening test, and then the positive samples could be randomly tested by MPN. PMID:25182727

  10. High resolution melting curve analysis, a rapid and affordable method for mutation analysis in childhood acute myeloid leukemia

    Directory of Open Access Journals (Sweden)

    Yin eLiu

    2014-09-01

    Full Text Available Background: Molecular genetic alterations with prognostic significance have been described in childhood acute myeloid leukemia (AML. The aim of this study was to establish cost-effective techniques to detect mutations of FMS-like tyrosine kinase 3 (FLT3, Nucleophosmin 1 (NPM1, and a partial tandem duplication within the mixed lineage leukemia (MLL-PTD genes in childhood AML. Procedure: Ninety-nine children with newly diagnosed AML were included in this study. We developed a fluoresent dye SYTO-82 based high resolution melting curve (HRM anaylsis to detect FLT3 internal tandem duplication (FLT3-ITD, FLT3 tyrosine kinase domain (FLT3-TKD and NPM1 mutations. MLL-PTD was screened by real-time quantitative PCR. Results: The HRM methodology correlated well with gold standard Sanger sequencing with less cost. Among the 99 patients studied, the FLT3-ITD mutation was associated with significantly worse event free survival (EFS. Patients with the NPM1 mutation had significantly better EFS and overall survival. However, HRM was not sensitive enough for minimal residual disease monitoring. Conclusions: HRM was a rapid and efficient method for screening of FLT3 and NPM1 gene mutations. It was both affordable and accurate, especially in resource underprivileged regions. Our results indicated that HRM could be a useful clinical tool for rapid and cost effective screening of the FLT3 and NPM1 mutations in AML patients.

  11. BRaf signaling principles unveiled by large-scale human mutation analysis with a rapid lentivirus-based gene replacement method.

    Science.gov (United States)

    Lim, Chae-Seok; Kang, Xi; Mirabella, Vincent; Zhang, Huaye; Bu, Qian; Araki, Yoichi; Hoang, Elizabeth T; Wang, Shiqiang; Shen, Ying; Choi, Sukwoo; Kaang, Bong-Kiun; Chang, Qiang; Pang, Zhiping P; Huganir, Richard L; Zhu, J Julius

    2017-03-15

    Rapid advances in genetics are linking mutations on genes to diseases at an exponential rate, yet characterizing the gene mutation-cell behavior relationships essential for precision medicine remains a daunting task. More than 350 mutations on small GTPase BRaf are associated with various tumors, and ∼40 mutations are associated with the neurodevelopmental disorder cardio-facio-cutaneous syndrome (CFC). We developed a fast cost-effective lentivirus-based rapid gene replacement method to interrogate the physiopathology of BRaf and ∼50 disease-linked BRaf mutants, including all CFC-linked mutants. Analysis of simultaneous multiple patch-clamp recordings from 6068 pairs of rat neurons with validation in additional mouse and human neurons and multiple learning tests from 1486 rats identified BRaf as the key missing signaling effector in the common synaptic NMDA-R-CaMKII-SynGap-Ras-BRaf-MEK-ERK transduction cascade. Moreover, the analysis creates the original big data unveiling three general features of BRaf signaling. This study establishes the first efficient procedure that permits large-scale functional analysis of human disease-linked mutations essential for precision medicine. © 2017 Lim et al.; Published by Cold Spring Harbor Laboratory Press.

  12. A rapid method for selecting suitable animal species for studying pathogen interactions with plasma protein ligands in vivo.

    Science.gov (United States)

    Naudin, Clément; Schumski, Ariane; Salo-Ahen, Outi M H; Herwald, Heiko; Smeds, Emanuel

    2017-05-01

    Species tropism constitutes a serious problem for developing relevant animal models of infection. Human pathogens can express virulence factors that show specific selectivity to human proteins, while their affinity for orthologs from other species can vary significantly. Suitable animal species must be used to analyse whether virulence factors are potential targets for drug development. We developed an assay that rapidly predicts applicable animal species for studying virulence factors binding plasma proteins. We used two well-characterized Staphylococcus aureus proteins, SSL7 and Efb, to develop an ELISA-based inhibition assay using plasma from different animal species. The interaction between SSL7 and human C5 and the binding of Efb to human fibrinogen and human C3 was studied. Affinity experiments and Western blot analyses were used to validate the assay. Human, monkey and cat plasma interfered with binding of SSL7 to human C5. Binding of Efb to human fibrinogen was blocked in human, monkey, gerbil and pig plasma, while human, monkey, gerbil, rabbit, cat and guinea pig plasma inhibited the binding of Efb to human C3. These results emphasize the importance of choosing correct animal models, and thus, our approach is a rapid and cost-effective method that can be used to prevent unnecessary animal experiments. © 2017 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  13. LC-ESI/MS/MS method for rapid screening and confirmation of 44 exogenous anabolic steroids in human urine.

    Science.gov (United States)

    Jeon, Byoung Wook; Yoo, Hye Hyun; Jeong, Eun Sook; Kim, Ho Jun; Jin, Changbae; Kim, Dong Hyun; Lee, Jaeick

    2011-09-01

    A sensitive and rapid method based on liquid chromatography-triple-quadrupole tandem mass spectrometry (LC-MS/MS) with electrospray ionization (ESI) has been developed and validated for the screening and confirmation of 44 exogenous anabolic steroids (29 parent steroids and 15 metabolites) in human urine. The method involves an enzymatic hydrolysis, liquid-liquid extraction, and detection by LC-MS/MS. A triple-quadrupole mass spectrometer was operated in positive ESI mode with selected reaction monitoring (SRM) mode for the screening and product ion scan mode for the confirmation. The protonated molecular ions were used as precursor ions for the SRM analysis and product ion scan. The intraday and interday precisions of the target analytes at concentrations of the minimum required performance levels for the screening were 2-14% and 2-15%, respectively. The limits of detection for the screening and confirmation method were 0.1-10 ng/mL and 0.2-10 ng/mL, respectively, for 44 steroids. This method was successfully applied to analysis of urine samples from suspected anabolic steroid abusers.

  14. Rapid screening of toxic salbutamol, ractopamine, and clenbuterol in pork sample by high-performance liquid chromatography—UV method

    Directory of Open Access Journals (Sweden)

    Kunping Yan

    2016-04-01

    Full Text Available A rapid and simple high-performance liquid chromatography–UV method was developed for the separation and quantification of salbutamol, ractopamine, and clenbuterol in pork. A mixture of acetonitrile–formic acid–ammonium acetate was used as the mobile phase to separate three β-agonists on a C18 column with gradient. The effects of the addition of formic acid and ammonium acetate to mobile phases on the separation of β-agonists were investigated. These additives can greatly improve the resolution and sensitivity. Under the optimized chromatographic condition, this separation does not need extra sample preparation. Complete baseline separation of three β-agonists was achieved in 0.99. Excellent method reproducibility was found by intra- and interday precisions with a relative standard deviation of < 3%. The detection limit (S/N = 3 was found to be <0.05 μg/L; this method can be used for routine screening of the β-agonist residues in foods of animal origin before being identified by confirmatory methods.

  15. Simple and rapid high-performance liquid chromatographic method for the determination of aspartame and its metabolites in foods.

    Science.gov (United States)

    Gibbs, B F; Alli, I; Mulligan, C N

    1996-02-23

    A method for the determination of aspartame (N-L-alpha-aspartyl-L-phenylalanine methyl ester) and its metabolites, applicable on a routine quality assurance basis, is described. Liquid samples (diet Coke, 7-Up, Pepsi, etc.) were injected directly onto a mini-cartridge reversed-phase column on a high-performance liquid chromatographic system, whereas solid samples (Equal, hot chocolate powder, pudding, etc.) were extracted with water. Optimising chromatographic conditions resulted in resolved components of interest within 12 min. The by-products were confirmed by mass spectrometry. Although the method was developed on a two-pump HPLC system fitted with a diode-array detector, it is straightforward and can be transformed to the simplest HPLC configuration. Using a single-piston pump (with damper), a fixed-wavelength detector and a recorder/integrator, the degradation of products can be monitored as they decompose. The results obtained were in harmony with previously reported tedious methods. The method is simple, rapid, quantitative and does not involve complex, hazardous or toxic chemistry.

  16. A novel UHPLC method for the rapid and simultaneous determination of daidzein, genistein and equol in human urine.

    Science.gov (United States)

    Redruello, Begoña; Guadamuro, Lucía; Cuesta, Isabel; Álvarez-Buylla, Jorge R; Mayo, Baltasar; Delgado, Susana

    2015-11-15

    This work reports on a novel method involving reverse-phased ultra-high performance liquid chromatography (UHPLC) plus a spectrophotometric photodiode array/fluorescence (FLR) detection system for determining the concentration of equol and major soy isoflavones (daidzein and genistein) in human urine. The proposed method was validated in terms of its linearity, sensitivity, accuracy (recovery) and precision (intra- and inter-day repeatability). The isoflavone profiles of urine samples from a group of menopausal women following oral soy isoflavone supplementation were determined and compared. Screening for equol-producer status was accomplished with high sensitivity (detection limit of the FLR detector 2.93nM). The method involves a short chromatographic run time compared to conventional HPLC methods while allowing for the simultaneous and reliable quantification of daidzein, genistein and equol in human urine. It also allows for the rapid screening of multiple urine samples when testing for equol production status and checking patient adherence to isoflavone treatment regimens. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Rapid identification of Mycobacterium tuberculosis infection by a new array format-based surface plasmon resonance method

    Science.gov (United States)

    Hsieh, Shang-Chen; Chang, Chia-Chen; Lu, Chia-Chen; Wei, Chia-Fong; Lin, Chuan-Sheng; Lai, Hsin-Chih; Lin, Chii-Wann

    2012-03-01

    Tubercle bacillus [TB] is one of the most important chronic infectious diseases that cause millions of deaths annually. While conventional smear microscopy and culture methods are widely used for diagnosis of TB, the former is insensitive, and the latter takes up to 6 to 8 weeks to provide a result, limiting the value of these methods in aiding diagnosis and intermediate decisions on treatment. Therefore, a rapid detection method is essential for the diagnosis, prognosis assessment, and recurrence monitoring. A new surface plasmon resonance [SPR] biosensor based on an array format, which allowed immobilizing nine TB antigens onto the sensor chip, was constructed. Simultaneous determination of multiple TB antibodies in serum had been accomplished with this array-based SPR system. The results were compared with enzyme-linked immunosorbent assay, a conventional immunological method. Array-based SPR showed more advantages in providing label-free and real-time detection. Additionally, the high sensitivity and specificity for the detection of TB infection showed its potential for future development of biosensor arrays for TB diagnosis.

  18. Presenting a rapid method for detection of Bacillus cereus, Listeria monocytogenes and Campylobacter jejuni in food samples

    Directory of Open Access Journals (Sweden)

    Ali Razei

    2017-08-01

    Full Text Available Objective(s: Listeria monocytogens, Bacillus cereus and Campylobacter jejuni are three toxin producing bacteria over the world, especially in Iran, and it is essential to find a certain, rapid procedure to identify these microorganisms. In this research, these bacteria were simultaneously detected by multiplex PCR technique in foods. Materials and Methods: The primary approval of bacterial strains was performed by biochemical tests. PCR primers were designed based on the nucleotide sequences of the NHEB/NHEC gene of B. cereus, the hly gene of L. monocytogenes and the C gene of C. jejuni. The specificity of Multiplex PCR method was determined using seven food poisoning bacteria including Salmonella typhi, Shigella dysentery, Yersinia pestis, Staphylococcus aureus, Clostridium perfringens, Clostridium botulinum and Vibrio cholerae. To confirm the reaction, DNA extraction was performed from 30 food samples (milk, and gene amplification was performed by PCR. The length of amplified fragments was 300 bp, 210 bp and 160 bpfor NHEB/NHEC, hly and C genes, respectively. Results: The detection limits of the PCR method were 5, 4 and 3 pg for L. monocytogenes, B. cereus and C. jejuni, respectively. Specifisity test showed that this reaction is spesific to these 3 bacteria. Conclusion: In this study, we  introduced a new multiplex PCR method for simultsnus detection of L. monocytogens, B. cereus and C. jejuni. These results can be used  for detection of other toxin producing bacteria in food.

  19. Invert biopanning: A novel method for efficient and rapid isolation of scFvs by phage display technology.

    Science.gov (United States)

    Rahbarnia, Leila; Farajnia, Safar; Babaei, Hossein; Majidi, Jafar; Veisi, Kamal; Tanomand, Asghar; Akbari, Bahman

    2016-11-01

    Phage display is a prominent screening technique for development of novel high affinity antibodies against almost any antigen. However, removing false positive clones in screening process remains a challenge. The aim of this study was to develop an efficient and rapid method for isolation of high affinity scFvs by removing NSBs without losing rare specific clones. Therefore, a novel two rounds strategy called invert biopanning was developed for isolating high affinity scFvs against EGFRvIII antigen from human scFv library. The efficiency of invert biopanning method (procedure III) was analyzed by comparing with results of conventional biopanning methods (procedures I and II). According to the results of polyclonal ELISA, the second round of procedure III displayed highest binding affinity against EGFRvIII peptide accompanied by lowest NSB comparing to other two procedures. Several positive clones were identified among output phages of procedure III by monoclonal phage ELISA which displayed high affinity to EGFRvIII antigen. In conclusion, results of our study indicate that invert biopanning is an efficient method for avoiding NSBs and conservation of rare specific clones during screening of a scFv phage library. Novel anti EGFRvIII scFv isolated could be a promising candidate for potential use in treatment of EGFRvIII expressing cancers. Copyright © 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  20. Rapid and sensitive liquid chromatography-tandem mass spectrometry method for the quantitation of levodropropizine in human plasma.

    Science.gov (United States)

    Tang, Yunbiao; Zhao, Limei; Wang, Yingwu; Fawcett, J Paul; Gu, Jingkai

    2005-05-05

    A rapid and sensitive LC-MS-MS method for quantifying levodropropizine in human plasma after oral administration of a single-dose (60 mg/day) was developed and validated. The sample preparation used liquid-liquid extraction with a mixture of dichloromethane-diethyl ether (2:3, v/v) in a basic environment. The retention time of levodropropizne and zolmitriptan (used as internal standard) was 1.6 and 1.4 min, respectively. The assay was linear over the range 0.25-500 ng/mL with a LOQ of 0.25 ng/mL. The intra- and inter-day precision were levodropropizine concentration profile in human plasma was determined.