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Sample records for acid detection methods

  1. Nucleic Acid Detection Methods

    DOEpatents

    Smith, Cassandra L.; Yaar, Ron; Szafranski, Przemyslaw; Cantor, Charles R.

    1998-05-19

    The invention relates to methods for rapidly determining the sequence and/or length a target sequence. The target sequence may be a series of known or unknown repeat sequences which are hybridized to an array of probes. The hybridized array is digested with a single-strand nuclease and free 3'-hydroxyl groups extended with a nucleic acid polymerase. Nuclease cleaved heteroduplexes can be easily distinguish from nuclease uncleaved heteroduplexes by differential labeling. Probes and target can be differentially labeled with detectable labels. Matched target can be detected by cleaving resulting loops from the hybridized target and creating free 3-hydroxyl groups. These groups are recognized and extended by polymerases added into the reaction system which also adds or releases one label into solution. Analysis of the resulting products using either solid phase or solution. These methods can be used to detect characteristic nucleic acid sequences, to determine target sequence and to screen for genetic defects and disorders. Assays can be conducted on solid surfaces allowing for multiple reactions to be conducted in parallel and, if desired, automated.

  2. Nucleic acid detection methods

    DOEpatents

    Smith, C.L.; Yaar, R.; Szafranski, P.; Cantor, C.R.

    1998-05-19

    The invention relates to methods for rapidly determining the sequence and/or length a target sequence. The target sequence may be a series of known or unknown repeat sequences which are hybridized to an array of probes. The hybridized array is digested with a single-strand nuclease and free 3{prime}-hydroxyl groups extended with a nucleic acid polymerase. Nuclease cleaved heteroduplexes can be easily distinguish from nuclease uncleaved heteroduplexes by differential labeling. Probes and target can be differentially labeled with detectable labels. Matched target can be detected by cleaving resulting loops from the hybridized target and creating free 3-hydroxyl groups. These groups are recognized and extended by polymerases added into the reaction system which also adds or releases one label into solution. Analysis of the resulting products using either solid phase or solution. These methods can be used to detect characteristic nucleic acid sequences, to determine target sequence and to screen for genetic defects and disorders. Assays can be conducted on solid surfaces allowing for multiple reactions to be conducted in parallel and, if desired, automated. 18 figs.

  3. Nucleic acid detection system and method for detecting influenza

    DOEpatents

    Cai, Hong; Song, Jian

    2015-03-17

    The invention provides a rapid, sensitive and specific nucleic acid detection system which utilizes isothermal nucleic acid amplification in combination with a lateral flow chromatographic device, or DNA dipstick, for DNA-hybridization detection. The system of the invention requires no complex instrumentation or electronic hardware, and provides a low cost nucleic acid detection system suitable for highly sensitive pathogen detection. Hybridization to single-stranded DNA amplification products using the system of the invention provides a sensitive and specific means by which assays can be multiplexed for the detection of multiple target sequences.

  4. Advances in nucleic acid-based detection methods.

    PubMed Central

    Wolcott, M J

    1992-01-01

    Laboratory techniques based on nucleic acid methods have increased in popularity over the last decade with clinical microbiologists and other laboratory scientists who are concerned with the diagnosis of infectious agents. This increase in popularity is a result primarily of advances made in nucleic acid amplification and detection techniques. Polymerase chain reaction, the original nucleic acid amplification technique, changed the way many people viewed and used nucleic acid techniques in clinical settings. After the potential of polymerase chain reaction became apparent, other methods of nucleic acid amplification and detection were developed. These alternative nucleic acid amplification methods may become serious contenders for application to routine laboratory analyses. This review presents some background information on nucleic acid analyses that might be used in clinical and anatomical laboratories and describes some recent advances in the amplification and detection of nucleic acids. PMID:1423216

  5. Methods for point-of-care detection of nucleic acid in a sample

    DOEpatents

    Bearinger, Jane P.; Dugan, Lawrence C.

    2015-12-29

    Provided herein are methods and apparatus for detecting a target nucleic acid in a sample and related methods and apparatus for diagnosing a condition in an individual. The condition is associated with presence of nucleic acid produced by certain pathogens in the individual.

  6. A Nucleic Acid Probe and Method for the Detection of Shigella and Enteroinvasive E. coli Bacteria.

    DTIC Science & Technology

    This invention relates to nucleic acid probes and a method for the rapid detection of Shigella and enteroinvasive Escherichia coli, the causative agents of bacterial dysentery, by use of a nucleic acid hybridization probe, equivalent to a plasmid DNA region encoding one of 4 specific invasion-associated, peptides of all strains of Shigella and enterinvasive E . coli , in a nucleic acid hybridization reaction with a clinical specimen containing dysentery bacteria.

  7. Nucleic acid detection kits

    DOEpatents

    Hall, Jeff G.; Lyamichev, Victor I.; Mast, Andrea L.; Brow, Mary Ann; Kwiatkowski, Robert W.; Vavra, Stephanie H.

    2005-03-29

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based on charge. The present invention also provides methods for the detection of non-target cleavage products via the formation of a complete and activated protein binding region. The invention further provides sensitive and specific methods for the detection of nucleic acid from various viruses in a sample.

  8. Capillary electrophoresis method with UV-detection for analysis of free amino acids concentrations in food.

    PubMed

    Omar, Mei Musa Ali; Elbashir, Abdalla Ahmed; Schmitz, Oliver J

    2017-01-01

    Simple and inexpensive capillary electrophoresis with UV-detection method (CE-UV) was optimized and validated for determination of six amino acids namely (alanine, asparagine, glutamine, proline, serine and valine) for Sudanese food. Amino acids in the samples were derivatized with 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl) prior to CE-UV analysis. Labeling reaction conditions (100mM borate buffer at pH 8.5, labeling reaction time 60min, temperature 70°C and NBD-Cl concentration 40mM) were systematically investigated. The optimal conditions for the separation were 100mM borate buffer at pH 9.7 and detected at 475nm. The method was validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ), precision (repeatability) (RSD%) and accuracy (recovery). Good linearity was achieved for all amino acids (r(2)>0.9981) in the concentration range of 2.5-40mg/L. The LODs in the range of 0.32-0.56mg/L were obtained. Recoveries of amino acids ranging from 85% to 108%, (n=3) were obtained. The validated method was successfully applied for the determination of amino acids for Sudanese food samples.

  9. Method for rapid detection and identification of chaetomium and evaluation of resistance to peracetic acid.

    PubMed

    Nakayama, Motokazu; Hosoya, Kouichi; Tomiyama, Daisuke; Tsugukuni, Takashi; Matsuzawa, Tetsuhiro; Imanishi, Yumi; Yaguchi, Takashi

    2013-06-01

    In the beverage industry, peracetic acid has been increasingly used as a disinfectant for the filling machinery and environment due to merits of leaving no residue, it is safe for humans, and its antiseptic effect against fungi and endospores of bacteria. Recently, Chaetomium globosum and Chaetomium funicola were reported resistant to peracetic acid; however, little is known concerning the detail of peracetic acid resistance. Therefore, we assessed the peracetic acid resistance of the species of Chaetomium and related genera under identical conditions and made a thorough observation of the microstructure of their ascospores by transmission electron microscopy. The results of analyses revealed that C. globosum and C. funicola showed the high resistance to peracetic acid (a 1-D antiseptic effect after 900 s and 3-D antiseptic effect after 900 s) and had thick cell walls of ascospores that can impede the action mechanism of peracetic acid. We also developed specific primers to detect the C. globosum clade and identify C. funicola by using PCR to amplify the β-tubulin gene. PCR with the primer sets designed for C. globosum (Chae 4F/4R) and C. funicola (Cfu 2F/2R) amplified PCR products specific for the C. globosum clade and C. funicola, respectively. PCR with these two primer sets did not detect other fungi involved in food spoilage and environmental contamination. This detection and identification method is rapid and simple, with extremely high specificity.

  10. Nucleic acid detection compositions

    DOEpatents

    Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow, Mary Ann; Dahlberg, James L.

    2008-08-05

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

  11. Nucleic acid detection assays

    DOEpatents

    Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow, Mary Ann; Dahlberg, James E.

    2005-04-05

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

  12. Isothermal Amplification Methods for the Detection of Nucleic Acids in Microfluidic Devices

    PubMed Central

    Zanoli, Laura Maria; Spoto, Giuseppe

    2012-01-01

    Diagnostic tools for biomolecular detection need to fulfill specific requirements in terms of sensitivity, selectivity and high-throughput in order to widen their applicability and to minimize the cost of the assay. The nucleic acid amplification is a key step in DNA detection assays. It contributes to improving the assay sensitivity by enabling the detection of a limited number of target molecules. The use of microfluidic devices to miniaturize amplification protocols reduces the required sample volume and the analysis times and offers new possibilities for the process automation and integration in one single device. The vast majority of miniaturized systems for nucleic acid analysis exploit the polymerase chain reaction (PCR) amplification method, which requires repeated cycles of three or two temperature-dependent steps during the amplification of the nucleic acid target sequence. In contrast, low temperature isothermal amplification methods have no need for thermal cycling thus requiring simplified microfluidic device features. Here, the use of miniaturized analysis systems using isothermal amplification reactions for the nucleic acid amplification will be discussed. PMID:25587397

  13. Rapid detection and identification of beer-spoilage lactic acid bacteria by microcolony method.

    PubMed

    Asano, Shizuka; Iijima, Kazumaru; Suzuki, Koji; Motoyama, Yasuo; Ogata, Tomoo; Kitagawa, Yasushi

    2009-08-01

    We evaluated a microcolony method for the detection and identification of beer-spoilage lactic acid bacteria (LAB). In this approach, bacterial cells were trapped on a polycarbonate membrane filter and cultured on ABD medium, a medium that allows highly specific detection of beer-spoilage LAB strains. After short-time incubation, viable cells forming microcolonies were stained with carboxyfluorescein diacetate (CFDA) and counted with muFinder Inspection System. In our study, we first investigated the growth behavior of various beer-spoilage LAB by traditional culture method, and Lactobacillus lindneri and several L. paracollinoides strains were selected as slow growers on ABD medium. Then the detection speeds were evaluated by microcolony method, using these slowly growing strains. As a result, all of the slowly growing beer-spoilage LAB strains were detected within 3 days of incubation. The specificity of this method was found to be exceptionally high and even discriminated intra-species differences in beer-spoilage ability of LAB strains upon detection. These results indicate that our microcolony approach allows rapid and specific detection of beer-spoilage LAB strains with inexpensive CFDA staining. For further confirmation of species status of detected strains, subsequent treatment with species-specific fluorescence in situ hybridization (FISH) probes was shown as effective for identifying the CFDA-detected microcolonies to the species level. In addition, no false-positive results arising from noise signals were recognized for CFDA staining and FISH methods. Taken together, the developed microcolony method was demonstrated as a rapid and highly specific countermeasure against beer-spoilage LAB, and compared favorably with the conventional culture methods.

  14. Isomers/enantiomers of perfluorocarboxylic acids: Method development and detection in environmental samples

    EPA Science Inventory

    Perfluoroalkyl substances are globally distributed in both urban and remote settings, and routinely are detected in wildlife, humans, and the environment. One of the most prominent and routinely detected perfluoroalkyl substances is perfluorooctanoic acid (PFOA), which has been s...

  15. An HPLC method with UV detection, pH control, and reductive ascorbic acid for cyanuric acid analysis in water.

    PubMed

    Cantú, R; Evans, O; Kawahara, F K; Shoemaker, J A; Dufour, A P

    2000-12-01

    Every year over 250 million pounds of cyanuric acid (CA) and chlorinated isocyanurates are produced industrially. These compounds are standard ingredients in formulations for household bleaches, industrial cleansers, dishwasher compounds, general sanitizers, and chlorine stabilizers. The method developed for CA using high-performance liquid chromatography (HPLC) with UV detection simplifies and optimizes certain parameters of previous methodologies by effective pH control of the eluent (95% phosphate buffer: 5% methanol, v/v) to the narrow pH range of 7.2-7.4. UV detection was set at the optimum wavelength of 213 nm where the cyanuric ion absorbs strongly. Analysis at the lower pH range of 6.8-7.1 proved inadequate due to CA keto-enol tautomerism, while at pHs of <6.8 there were substantial losses in analytical sensitivity. In contrast, pHs of >7.4 proved more sensitive but their use was rejected because of CA elution at the chromatographic void volume and due to chemical interferences. The complex equilibria of chlorinated isocyanurates and associated species were suppressed by using reductive ascorbic acid to restrict the products to CA. UV, HPLC-UV, and electrospray ionization mass spectrometry techniques were combined to monitor the reactive chlorinated isocyanurates and to support the use of ascorbic acid. The resulting method is reproducible and measures CA in the 0.5-125 mg/L linear concentration range with a method detection limit of 0.05 mg/L in water.

  16. A rapid method for detecting protein-nucleic acid interactions by protein induced fluorescence enhancement

    PubMed Central

    Valuchova, Sona; Fulnecek, Jaroslav; Petrov, Alexander P.; Tripsianes, Konstantinos; Riha, Karel

    2016-01-01

    Many fundamental biological processes depend on intricate networks of interactions between proteins and nucleic acids and a quantitative description of these interactions is important for understanding cellular mechanisms governing DNA replication, transcription, or translation. Here we present a versatile method for rapid and quantitative assessment of protein/nucleic acid (NA) interactions. This method is based on protein induced fluorescence enhancement (PIFE), a phenomenon whereby protein binding increases the fluorescence of Cy3-like dyes. PIFE has mainly been used in single molecule studies to detect protein association with DNA or RNA. Here we applied PIFE for steady state quantification of protein/NA interactions by using microwell plate fluorescence readers (mwPIFE). We demonstrate the general applicability of mwPIFE for examining various aspects of protein/DNA interactions with examples from the restriction enzyme BamHI, and the DNA repair complexes Ku and XPF/ERCC1. These include determination of sequence and structure binding specificities, dissociation constants, detection of weak interactions, and the ability of a protein to translocate along DNA. mwPIFE represents an easy and high throughput method that does not require protein labeling and can be applied to a wide range of applications involving protein/NA interactions. PMID:28008962

  17. Isomers/enantiomers of perfluorocarboxylic acids: Method development and detection in environmental samples.

    PubMed

    Naile, Jonathan E; Garrison, A Wayne; Avants, Jimmy K; Washington, John W

    2016-02-01

    Perfluoroalkyl substances are globally distributed in both urban and remote settings, and routinely are detected in wildlife, humans, and the environment. One of the most prominent and routinely detected perfluoroalkyl substances is perfluorooctanoic acid (PFOA), which has been shown to be toxic to both humans and animals. PFOA exists as both linear and branched isomers; some of the branched isomers are chiral. A novel GC-NCI-MS method was developed to allow for isomer/enantiomer separation, which was achieved using two columns working in tandem; a 30-m DB-5MS column and a 30-m BGB-172 Analytik column. Samples were derivatized with diazomethane to form methyl esters of the PFOA isomers. In standards, at least eight PFOA isomers were detected, of which at least four were enantiomers of chiral isomers; one chiral isomer (P3) was sufficiently separated to allow for enantiomer-fraction calculations. Soil, sediment and plant samples from contaminated locations in Alabama and Georgia were analyzed. P3 was observed in most of these environmental samples, and was non-racemic in at least one sediment, suggesting the possibility of chirally selective generation from precursors or enantioselective sorption. In addition, the ratio of P3/linear PFOA was inversely related to distance from source, which we suggest might reflect a higher sorption affinity for the P3 over the linear isomer. This method focuses on PFOA, but preliminary results suggest that it should be broadly applicable to other chiral and achiral perfluorocarboxylic acids (PFCAs); e.g., we detected several other homologous PFCA isomers in our PFCA standards and some environmental samples.

  18. Methods for detecting ATP hydrolysis and nucleic acid unwinding of Japanese encephalitis virus NS3 helicase.

    PubMed

    Fang, Jin'e; Li, Huan; Peng, Guiqing; Cao, Shengbo; Zhen, F Fu; Chen, Huanchun; Song, Yunfeng

    2013-12-01

    Japanese encephalitis virus (JEV) is a mosquito-borne zoonotic pathogen that is prevalent in south-east Asia. Because there is no specific antiviral agent, JEV still causes a high rate of neurologic sequelae and mortality in humans. The helicase encoded by the NS3 gene of JEV has emerged recently as a novel antiviral target for treatment. In this study, a soluble recombinant JEV helicase protein was expressed and purified. Methods for detecting the ATP hydrolysis and nucleic acid unwinding activity were developed by luminescence and fluorescence resonance energy transfer (FRET). The concentrations of enzyme, substrate, capture strand, ATP, and divalent ions were optimised in the ATPase and helicase reactions. The feasibility of using these two methods for high-throughput screening of NS3 helicase inhibitors is discussed.

  19. 78 FR 16513 - Application of Advances in Nucleic Acid and Protein Based Detection Methods to Multiplex...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-15

    ... Detection Methods to Multiplex Detection of Transfusion-Transmissible Agents and Blood Cell Antigens in...- Transmissible Agents and Blood Cell Antigens in Blood Donations.'' The purpose of this public workshop is to... and blood cell antigen typing. The public workshop has been planned in partnership with the...

  20. A Reliable and Inexpensive Method of Nucleic Acid Extraction for the PCR-Based Detection of Diverse Plant Pathogens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A reliable extraction method is described for the preparation of total nucleic acids from several plant genera for subsequent detection of plant pathogens by PCR-based techniques. By the combined use of a modified CTAB (cetyltrimethylammonium bromide) extraction protocol and a semi-automatic homogen...

  1. Fluorescence detection in Lab-on-a-chip systems using ultrafast nucleic acid amplification methods

    NASA Astrophysics Data System (ADS)

    Gransee, Rainer; Schneider, Tristan; Elyorgun, Deniz; Strobach, Xenia; Schunck, Tobias; Gatscha, Theresia; Höth, Julian

    2014-05-01

    Today, nucleic amplification plays a key role in modern molecular biology allowing fast and specific laboratory diagnostics testing. An ultrafast microfluidic module (allowing 30 polymeric chain reaction (PCR) cycles in 6 minutes) based on an oscillating fluid plug concept was previously developed[1]. This system allows the amplification of native genomic deoxyribonucleic acid molecules (DNA) even from whole blood samples but still lacks some functionality compared to commercial bench top systems. This work presents the actual status of the renewed and advanced system, permitting the automated optical detection of not only the fluid plug position but also fluorescence detection. The system uses light emitting diodes (LED) for illumination and a low cost CMOS web-camera for optical detection. Image data processing allows the automated process control of the overall system components. Therefore, the system enables the performance of rapid and robust nucleic acid amplifications together with the integration of real time measurement technology. This allows the amplification and simultaneous quantification of the DNA molecules. The possibility to integrate swift nucleic amplification and optical detection into complex sample-to-answer analysis platforms opens up new pathways towards fast and transportable low-cost point of care devices.

  2. Comparison of nucleic acid extraction methods for the detection of Mycoplasma pneumoniae.

    PubMed

    Thurman, Kathleen A; Cowart, Kelley C; Winchell, Jonas M

    2009-12-01

    Four nucleic acid extraction procedures (2 automated and 2 manual) were compared for their efficiency at isolating Mycoplasma pneumoniae DNA. Oropharyngeal swabs from healthy volunteers were spiked with varying amounts of M. pneumoniae, extracted, and tested using real-time polymerase chain reaction. Our data indicate that both automated extraction methods consistently outperform the manual procedures.

  3. A simple, rapid method of nucleic acid extraction without tissue homogenization for detecting viroids by hybridization and RT-PCR.

    PubMed

    Nakahara, K; Hataya, T; Uyeda, I

    1999-01-01

    A simple, rapid method of nucleic acid extraction on a microcentrifuge tube scale for detecting viroids is presented. Five distinct citrus viroids (CVds), chrysanthemum stunt viroid (CSVd), hop stunt viroid (HSVd), hop latent viroid (HLVd) and potato spindle tuber viroid (PSTVd) were detected in their natural host plants by hybridization using cRNA probes and reverse transcription-polymerase chain reaction (RT-PCR). Nucleic acids (NA) were liberated from tissues by incubation in a buffer containing potassium ethyl xanthogenate (PEX) without tissue homogenization, and then precipitated with ethanol (NA-PEX). All the viroids except CVd-IV could be detected clearly in NA-PEX by hybridization. HSVd, HLVd and PSTVd could also be detected in NA-PEX by RT-PCR. Although CVds and CSVd could not be detected in NA-PEX by RT-PCR, they were detected after further purification: differential precipitation with 2-butoxyethanol and HCl treatment followed by ethanol-precipitation. In addition, PCR in the presence of tetramethylammonium chloride specifically amplified the cDNA of all five distinct CVds under the same temperature and cycle conditions. Since all the viroids could be detected in NA liberated by PEX, the amount of NA extracted by the method described here is sufficient for detecting viroids, enabling the processing of a large number of samples.

  4. Method for colorimetric detection of double-stranded nucleic acid using leuco triphenylmethane dyes.

    PubMed

    Miyamoto, Shigehiko; Sano, Sotaro; Takahashi, Koji; Jikihara, Takaaki

    2015-03-15

    Because loop-mediated isothermal amplification (LAMP) can amplify substantial amounts of DNA under isothermal conditions, its applications for simple genetic testing have attracted considerable attention. A positive LAMP reaction is indicated by the turbidity caused by by-products or by the color change after adding a metallochromic indicator to the reaction solution, but these methods have certain limitations. Leuco crystal violet (LCV), a colorless dye obtained after sodium sulfite treatment of crystal violet (CV), was used as a new colorimetric method for detecting LAMP. LCV is reconverted into CV through contact with double-stranded DNA (dsDNA). Therefore, the positive reaction of LAMP is indicated by color change from colorless to violet. The assay is sensitive enough to detect LAMP products, with a detection limit of 7.1 ng/μl for dsDNA. It is also highly selective to dsDNA, and interference with single-stranded DNA and deoxynucleotide triphosphates (dNTPs) is not observed. LCV facilitates direct colorimetric detection of the main product rather than a by-product of the LAMP reaction; therefore, this method can be used under various reaction conditions such as those with added pyrophosphatase in solution. This colorimetric LAMP detection method using LCV is useful for point-of-care genetic testing given its simplicity.

  5. Validation of the TaqMan Influenza A Detection Kit and a rapid automated total nucleic acid extraction method to detect influenza A virus in nasopharyngeal specimens.

    PubMed

    Bolotin, Shelly; De Lima, Cedric; Choi, Kam-Wing; Lombos, Ernesto; Burton, Laura; Mazzulli, Tony; Drews, Steven J

    2009-01-01

    This study describes the validation of the TaqMan Influenza A Detection Kit v2.0 combined with an automated nucleic acid extraction method. The limit of detection of this assay was determined by probit regression (95% confidence interval) to be 2 influenza A/PR/8/34 (H1N1) virus particles per microlitre. One hundred and eleven specimens previously tested using the Seeplex RV assay and viral culture methods were tested using the TaqMan Influenza A Detection Kit. Compared to the aggregate gold-standard, the sensitivity and specificity of the TaqMan Influenza A Detection Kit were 100% (35/35) and 97% (74/76), respectively. Because of its accuracy, quick turn-around-time and lyophilized bead form, the TaqMan Influenza A Detection Kit, combined with the NucliSense easyMAG automated extraction method, constitutes a reliable protocol for influenza A diagnosis.

  6. Method for the detection of specific nucleic acid sequences by polymerase nucleotide incorporation

    DOEpatents

    Castro, Alonso

    2004-06-01

    A method for rapid and efficient detection of a target DNA or RNA sequence is provided. A primer having a 3'-hydroxyl group at one end and having a sequence of nucleotides sufficiently homologous with an identifying sequence of nucleotides in the target DNA is selected. The primer is hybridized to the identifying sequence of nucleotides on the DNA or RNA sequence and a reporter molecule is synthesized on the target sequence by progressively binding complementary nucleotides to the primer, where the complementary nucleotides include nucleotides labeled with a fluorophore. Fluorescence emitted by fluorophores on single reporter molecules is detected to identify the target DNA or RNA sequence.

  7. An improved method for the immunological detection of mineral bound protein using hydrofluoric acid and direct capture.

    PubMed

    Craig, O E; Collins, M J

    2000-03-06

    Immunological detection of proteins adsorbed to mineral and ceramic surfaces has proved not only difficult but controversial. Unlike the immunological detection of proteins associated with carbonate or phosphate minerals (e.g. shells and bones) proteins adsorbed to siliceous minerals cannot readily be removed by dissolution of the mineral phase. We have previously examined alternative extraction methodologies which claim to bring the protein into solution, but found none of these to be effective. Here we report a novel strategy for immuno-detection of proteins adsorbed to siliceous minerals, the Digestion and Capture Immunoassay (DACIA). The method involves the use of cold, concentrated (4M) hydrofluoric acid (HF) with the simultaneous capture of liberated protein onto a solid phase. The combination of low temperatures and surface stabilisation enables us to detect epitopes from even partially degraded proteins. The method may have a wide application in forensic, archaeological, soil and earth sciences.

  8. A validated HPLC method with electrochemical detection for simultaneous assay of 5-aminosalicylic acid and its metabolite in human plasma.

    PubMed

    Palumbo, Giancarlo; Bacchi, Simona; Primavera, Luisa; Palumbo, Paola; Carlucci, Giuseppe

    2005-06-01

    A high-performance liquid chromatographic method was developed, validated and applied to the simultaneous determination of 5-aminosalicylic acid (5-ASA) and its acetylated metabolite (acetyl-5-ASA) in human plasma. The method involves liquid-liquid extraction with methanol followed by isocratic reversed-phase chromatography on a Kromasil KR100 C(18) column with electrochemical detection. The recovery, selectivity, linearity, precision and accuracy of the method were evaluated from spiked human plasma samples. The effects of mobile phase composition, buffer concentration, mobile phase pH and concentration of organic modifiers on retention of 5-ASA, acetyl 5-ASA and internal standard were investigated. Limits' of detection were 5 ng/mL for 5-ASA and 10 ng/mL for acetyl-5-ASA, respectively. The method can be used for supporting therapeutical drug monitoring and pharmacokinetic studies.

  9. A novel, sensitive and label-free loop-mediated isothermal amplification detection method for nucleic acids using luminophore dyes.

    PubMed

    Roy, Sharmili; Wei, Sim Xiao; Ying, Jean Liew Zhi; Safavieh, Mohammadali; Ahmed, Minhaz Uddin

    2016-12-15

    Electrochemiluminescence (ECL) has been widely rendered for nucleic acid testing. Here, we integrate loop-mediated isothermal amplification (LAMP) with ECL technique for DNA detection and quantification. The target LAMP DNA bound electrostatically with [Ru(bpy)3](+2) on the carbon electrode surface, and an ECL reaction was triggered by tripropylamine (TPrA) to yield luminescence. We illustrated this method as a new and highly sensitive strategy for the detection of sequence-specific DNA from different meat species at picogram levels. The proposed strategy renders the signal amplification capacities of TPrA and combines LAMP with inherently high sensitivity of the ECL technique, to facilitate the detection of low quantities of DNA. By leveraging this technique, target DNA of Sus scrofa (pork) meat was detected as low as 1pg/µL (3.43×10(-1)copies/µL). In addition, the proposed technique was applied for detection of Bacillus subtilis DNA samples and detection limit of 10pg/µL (2.2×10(3)copies/µL) was achieved. The advantages of being isothermal, sensitive and robust with ability for multiplex detection of bio-analytes makes this method a facile and appealing sensing modality in hand-held devices to be used at the point-of-care (POC).

  10. Development of magnetic resonance imaging based detection methods for beta amyloids via sialic acid-functionalized magnetic nanoparticles

    NASA Astrophysics Data System (ADS)

    Kouyoumdjian, Hovig

    The development of a non-invasive method for the detection of Alzheimer's disease is of high current interest, which can be critical in early diagnosis and in guiding preventive treatment of the disease. The aggregates of beta amyloids are a pathological hallmark of Alzheimer's disease. Carbohydrates such as sialic acid terminated gangliosides have been shown to play significant roles in initiation of amyloid aggregation. Herein, we report a biomimetic approach using sialic acid coated iron oxide superparamagnetic nanoparticles for in vitro detection in addition to the assessment of the in vivo mouse-BBB (Blood brain barrier) crossing of the BSA (bovine serum albumin)-modified ones. The sialic acid functionalized dextran nanoparticles were shown to bind with beta amyloids through several techniques including ELISA (enzyme linked immunosorbent assay), MRI (magnetic resonance imaging), TEM (transmission electron microscopy), gel electrophoresis and tyrosine fluorescence assay. The superparamagnetic nature of the nanoparticles allowed easy detection of the beta amyloids in mouse brains in both in vitro and ex vivo model by magnetic resonance imaging. Furthermore, the sialic acid nanoparticles greatly reduced beta amyloid induced cytotoxicity to SH-SY5Y neuroblastoma cells, highlighting the potential of the glyconanoparticles for detection and imaging of beta amyloids. Sialic acid functionalized BSA (bovine serum albumin) nanoparticles also showed significant binding to beta amyloids, through ELISA and ex vivo mouse brain MRI experiments. Alternatively, the BBB crossing was demonstrated by several techniques such as confocal microscopy, endocytosis, exocytosis assays and were affirmed by nanoparticles transcytosis assays through bEnd.3 endothelial cells. Finally, the BBB crossing was confirmed by analyzing the MRI signal of nanoparticle-injected CD-1 mice.

  11. Optimization of the tartrate-resistant acid phosphatase detection by histochemical method

    PubMed Central

    Galvão, M.J.; Santos, A. R.; Ribeiro, M.D.; Ferreira, A.; Nolasco, F.

    2011-01-01

    According to the new kidney disease improving global outcomes (KDIGO) guidelines, the term of renal osteodystrophy, should be used exclusively in reference to the invasive diagnosis of bone abnormalities. Due to the low sensitivity and specificity of biochemical serum markers of bone remodelling, the performance of bone biopsies is highly stimulated in dialysis patients and after kidney transplantation. The tartrate-resistant acid phosphatase (TRACP) is an iso-enzyme of the group of acid phosphatases, which is highly expressed by activated osteoclasts and macrophages. TRACP in osteoclasts is in intracytoplasmic vesicles that transport the products of bone matrix degradation. Being present in activated osteoclasts, the identification of this enzyme by histochemistry in undecalcified bone biopsies is an excellent method to quantify the resorption of bone. Since it is an enzymatic histochemical method for a thermolabile enzyme, the temperature at which it is performed is particularly relevant. This study aimed to determine the optimal temperature for identification of TRACP in activated osteoclasts in undecalcified bone biopsies embedded in methylmethacrylate. We selected 10 cases of undecalcified bone biopsies from hemodialysis patients with the diagnosis of secondary hyperparathyroidism. Sections of 5 µm were stained to identify TRACP at different incubation temperatures (37°, 45°, 60°, 70° and 80°C) for 30 minutes. Activated osteoclasts stained red and trabecular bone (mineralized bone) was contrasted with toluidine blue. This approach also increased the visibility of the trabecular bone resorption areas (Howship lacunae). Unlike what is suggested in the literature and in several international protocols, we found that the best results were obtained with temperatures between 60°C and 70°C. For technical reasons and according to the results of the present study, we recommended that, for an incubation time of 30 min, the reaction should be carried out at 60

  12. AN HPLC METHOD WITH UV DETECTION, PH CONTROL, AND REDUCTIVE ASCORBIC ACID FOR CYANURIC ACID ANALYSIS IN WATER

    EPA Science Inventory

    Every year over 250 million pounds of cyanuric acid (CA) and chloroisocyanurates are produced industrially. These compounds are standard ingredients in formulations for household bleaches, industrial cleansers, dishwasher compounds, general sanitizers, and chlorine stabilizers. ...

  13. Evaluation of Molecular Methods for the Detection and Quantification of Pathogen-Derived Nucleic Acids in Sediment

    PubMed Central

    Farkas, Kata; Hassard, Francis; McDonald, James E.; Malham, Shelagh K.; Jones, Davey L.

    2017-01-01

    The accurate detection of pathogens in environmental matrices, such as sediment, is critical in understanding pathogen fate and behavior in the environment. In this study, we assessed the usefulness of methods for the detection and quantification of Vibrio spp. and norovirus (NoV) nucleic acids in sediment. For bacteria, a commonly used direct method using hexadecyltrimethylammonium bromide (CTAB) and phenol-chloroform-isoamyl alcohol (PCI) extraction was optimized, whereas for NoV, direct and indirect (virus elution—concentration) methods were evaluated. For quantification, commercially available quantitative PCR (qPCR) and reverse transcription qPCR (RT-qPCR) kits were tested alongside a digital PCR (dPCR) approach. CTAB-based extraction combined with 16 h polyethylene glycol 6000 (PEG6000) precipitation was found to be suitable for the direct extraction of high abundance bacterial and viral nucleic acids. For the indirect extraction of viral RNA, beef extract-based elution followed by PEG6000 precipitation and extraction using the NucliSENS® MiniMag® Nucleic Acid Purification System and the PowerViral® Environmental RNA/DNA Isolation Kit and qRT-PCR resulted in 83–112 and 63–69% recoveries of NoV, respectively. dPCR resulted in lower viral recoveries (47 and 9%) and ~4 orders of magnitude lower Vibrio concentrations (3.6–4.6 log10 gc/100 g sediment) than was observed using qPCR. The use of internal controls during viral quantification revealed that the RT step was more affected by inhibitors than the amplification. The methods described here are suitable for the enumeration of viral and/or bacterial pathogens in sediment, however the use of internal controls to assess efficiency is recommended. PMID:28174565

  14. Comparative evaluation of commercially available manual and automated nucleic acid extraction methods for rotavirus RNA detection in stools.

    PubMed

    Esona, Mathew D; McDonald, Sharla; Kamili, Shifaq; Kerin, Tara; Gautam, Rashi; Bowen, Michael D

    2013-12-01

    Rotaviruses are a major cause of viral gastroenteritis in children. For accurate and sensitive detection of rotavirus RNA from stool samples by reverse transcription-polymerase chain reaction (RT-PCR), the extraction process must be robust. However, some extraction methods may not remove the strong RT-PCR inhibitors known to be present in stool samples. The objective of this study was to evaluate and compare the performance of six extraction methods used commonly for extraction of rotavirus RNA from stool, which have never been formally evaluated: the MagNA Pure Compact, KingFisher Flex and NucliSENS easyMAG instruments, the NucliSENS miniMAG semi-automated system, and two manual purification kits, the QIAamp Viral RNA kit and a modified RNaid kit. Using each method, total nucleic acid or RNA was extracted from eight rotavirus-positive stool samples with enzyme immunoassay optical density (EIA OD) values ranging from 0.176 to 3.098. Extracts prepared using the MagNA Pure Compact instrument yielded the most consistent results by qRT-PCR and conventional RT-PCR. When extracts prepared from a dilution series were extracted by the 6 methods and tested, rotavirus RNA was detected in all samples by qRT-PCR but by conventional RT-PCR testing, only the MagNA Pure Compact and KingFisher Flex extracts were positive in all cases. RT-PCR inhibitors were detected in extracts produced with the QIAamp Viral RNA Mini kit. The findings of this study should prove useful for selection of extraction methods to be incorporated into future rotavirus detection and genotyping protocols.

  15. An Optimized Analytical Method for the Simultaneous Detection of Iodoform, Iodoacetic Acid, and Other Trihalomethanes and Haloacetic Acids in Drinking Water

    PubMed Central

    Jiang, Songhui; Templeton, Michael R.; He, Gengsheng; Qu, Weidong

    2013-01-01

    An optimized method is presented using liquid-liquid extraction and derivatization for the extraction of iodoacetic acid (IAA) and other haloacetic acids (HAA9) and direct extraction of iodoform (IF) and other trihalomethanes (THM4) from drinking water, followed by detection by gas chromatography with electron capture detection (GC-ECD). A Doehlert experimental design was performed to determine the optimum conditions for the five most significant factors in the derivatization step: namely, the volume and concentration of acidic methanol (optimized values  = 15%, 1 mL), the volume and concentration of Na2SO4 solution (129 g/L, 8.5 mL), and the volume of saturated NaHCO3 solution (1 mL). Also, derivatization time and temperature were optimized by a two-variable Doehlert design, resulting in the following optimized parameters: an extraction time of 11 minutes for IF and THM4 and 14 minutes for IAA and HAA9; mass of anhydrous Na2SO4 of 4 g for IF and THM4 and 16 g for IAA and HAA9; derivatization time of 160 min and temperature at 40°C. Under optimal conditions, the optimized procedure achieves excellent linearity (R2 ranges 0.9990–0.9998), low detection limits (0.0008–0.2 µg/L), low quantification limits (0.008–0.4 µg/L), and good recovery (86.6%–106.3%). Intra- and inter-day precision were less than 8.9% and 8.8%, respectively. The method was validated by applying it to the analysis of raw, flocculated, settled, and finished waters collected from a water treatment plant in China. PMID:23613747

  16. Development of a novel immunobiosensor method for the rapid detection of okadaic acid contamination in shellfish extracts.

    PubMed

    Llamas, Nuria M; Stewart, Linda; Fodey, Terry; Higgins, H Cowan; Velasco, María Luisa R; Botana, Luis M; Elliott, Christopher T

    2007-09-01

    The mouse bioassay is the methodology that is most widely used to detect okadaic acid (OA) in shellfish samples. This is one of the best-known toxins, and it belongs to the family of marine biotoxins referred to as the diarrhetic shellfish poisons (DSP). Due to animal welfare concerns, alternative methods of toxin detection are being sought. A rapid and specific biosensor immunoassay method was developed and validated for the detection of OA. An optical sensor instrument based on the surface plasmon resonance (SPR) phenomenon was utilised. A polyclonal antibody to OA was raised against OA-bovine thyroglobulin conjugate and OA-N-hydroxy succinimide ester was immobilised onto an amine sensor chip surface. The assay parameters selected for the analysis of the samples were: antibody dilution, 1/750; ratio of antibody to standard, 1:1; volume of sample injected, 25 microl min(-1); flow rate, 25 microl min(-1). An assay action limit of 126 ng g(-1) was established by analysing of 20 shellfish samples spiked with OA at the critical concentration of 160 ng g(-1), which is the action limit established by the European Union (EU). At this concentration of OA, the assay delivered coefficient of variations (CVs) of <10%. The chip surface developed was shown to be highly stable, allowing more than 50 analyses per channel. When the concentrations of OA determined with the biosensor method were compared with the values obtained by LC-MS in contaminated shellfish samples, the correlation between the two analytical methods was found to be highly satisfactory (r(2) = 0.991).

  17. Okadaic acid meet and greet: an insight into detection methods, response strategies and genotoxic effects in marine invertebrates.

    PubMed

    Prego-Faraldo, María Verónica; Valdiglesias, Vanessa; Méndez, Josefina; Eirín-López, José M

    2013-08-09

    Harmful Algal Blooms (HABs) constitute one of the most important sources of contamination in the oceans, producing high concentrations of potentially harmful biotoxins that are accumulated across the food chains. One such biotoxin, Okadaic Acid (OA), is produced by marine dinoflagellates and subsequently accumulated within the tissues of filtering marine organisms feeding on HABs, rapidly spreading to their predators in the food chain and eventually reaching human consumers causing Diarrhetic Shellfish Poisoning (DSP) syndrome. While numerous studies have thoroughly evaluated the effects of OA in mammals, the attention drawn to marine organisms in this regard has been scarce, even though they constitute primary targets for this biotoxin. With this in mind, the present work aimed to provide a timely and comprehensive insight into the current literature on the effect of OA in marine invertebrates, along with the strategies developed by these organisms to respond to its toxic effect together with the most important methods and techniques used for OA detection and evaluation.

  18. Locked Nucleic Acid Flow Cytometry-fluorescence in situ Hybridization (LNA flow-FISH): A Method for Bacterial Small RNA Detection

    DTIC Science & Technology

    2012-01-10

    Friedrich, U. & Lenke, J. Improved Enumeration of Lactic Acid Bacteria in Mesophilic Dairy Starter Cultures by Using Multiplex Quantitative Real...messenger RNA using locked nucleic acid probes. Anal. Biochem. 390, 109-114 (2009). 13. Waters, L. & Storz, G. Regulatory RNAs in bacteria . Cell. 136, 615...Video Article Locked Nucleic Acid Flow Cytometry-fluorescence in situ Hybridization (LNA flow-FISH): a Method for Bacterial Small RNA Detection Kelly

  19. Zymographic detection of cinnamic acid decarboxylase activity.

    PubMed

    Prim, Núria; Pastor, F I Javier; Diaz, Pilar

    2002-11-01

    The manuscript includes a concise description of a new, fast and simple method for detection of cinnamic acid decarboxylase activity. The method is based on a color shift caused a by pH change and may be an excellent procedure for large screenings of samples from natural sources, as it involves no complex sample processing or purification. The method developed can be used in preliminary approaches to biotransformation processes involving detection of hydroxycinnamic acid decarboxylase activity.

  20. Determination of rosmarinic acid in sage and borage leaves by high-performance liquid chromatography with different detection methods.

    PubMed

    Bandoniene, Donata; Murkovic, Michael; Venskutonis, Petras R

    2005-08-01

    Rosmarinic acid is separated and identified on the basis of high-performance liquid chromatography (HPLC)-UV-mass spectrometry data in 80% methanol in water extracts from the leaves of Salvia species (S. officinalis, S. glutinosa, S. aethiopis, S. sclarea, and Borago officinalis) as a dominant radical scavenger towards the 2,2'-diphenyl-1-picrylhydrazyl (DPPH*) stable radical in HPLC-DPPH* system. The content of rosmarinic acid in the plants is calibrated and quantitated from chromatograms obtained by UV detection at 280 nm. The concentration ranges from 13.3 to 47.3 mg of the phenolic acid per gram dried leaves of all plants is tested. S. glutinosa and S. sclarea have the highest concentration of rosmarinic acid. The amount of rosmarinic acid in borage leaves is similar compared with Salvia officinalis (15 mg/g). The HPLC-DPPH* system is calibrated for quantitative DPPH* scavenging assessment of rosmarinic acid. The results reveal excellent correlation (r2 = 0.98) between the rosmarinic acid concentration and antiradical activity.

  1. Okadaic Acid Meet and Greet: An Insight into Detection Methods, Response Strategies and Genotoxic Effects in Marine Invertebrates

    PubMed Central

    Prego-Faraldo, María Verónica; Valdiglesias, Vanessa; Méndez, Josefina; Eirín-López, José M.

    2013-01-01

    Harmful Algal Blooms (HABs) constitute one of the most important sources of contamination in the oceans, producing high concentrations of potentially harmful biotoxins that are accumulated across the food chains. One such biotoxin, Okadaic Acid (OA), is produced by marine dinoflagellates and subsequently accumulated within the tissues of filtering marine organisms feeding on HABs, rapidly spreading to their predators in the food chain and eventually reaching human consumers causing Diarrhetic Shellfish Poisoning (DSP) syndrome. While numerous studies have thoroughly evaluated the effects of OA in mammals, the attention drawn to marine organisms in this regard has been scarce, even though they constitute primary targets for this biotoxin. With this in mind, the present work aimed to provide a timely and comprehensive insight into the current literature on the effect of OA in marine invertebrates, along with the strategies developed by these organisms to respond to its toxic effect together with the most important methods and techniques used for OA detection and evaluation. PMID:23939476

  2. Comparison of automated nucleic acid extraction methods for the detection of cytomegalovirus DNA in fluids and tissues

    PubMed Central

    Waggoner, Jesse J.

    2014-01-01

    Testing for cytomegalovirus (CMV) DNA is increasingly being used for specimen types other than plasma or whole blood. However, few studies have investigated the performance of different nucleic acid extraction protocols in such specimens. In this study, CMV extraction using the Cell-free 1000 and Pathogen Complex 400 protocols on the QIAsymphony Sample Processing (SP) system were compared using bronchoalveolar lavage fluid (BAL), tissue samples, and urine. The QIAsymphonyAssay Set-up (AS) system was used to assemble reactions using artus CMV PCR reagents and amplification was carried out on the Rotor-Gene Q. Samples from 93 patients previously tested for CMV DNA and negative samples spiked with CMV AD-169 were used to evaluate assay performance. The Pathogen Complex 400 protocol yielded the following results: BAL, sensitivity 100% (33/33), specificity 87% (20/23); tissue, sensitivity 100% (25/25), specificity 100% (20/20); urine, sensitivity 100% (21/21), specificity 100% (20/20). Cell-free 1000 extraction gave comparable results for BAL and tissue, however, for urine, the sensitivity was 86% (18/21) and specimen quantitation was inaccurate. Comparative studies of different extraction protocols and DNA detection methods in body fluids and tissues are needed, as assays optimized for blood or plasma will not necessarily perform well on other specimen types. PMID:24765569

  3. Method for detecting biological toxins

    SciTech Connect

    Ligler, F.S.; Campbell, J.R.

    1992-01-01

    Biological toxins are indirectly detected by using polymerase chain reaction to amplify unique nucleic acid sequences coding for the toxins or enzymes unique to toxin synthesis. Buffer, primers coding for the unique nucleic acid sequences and an amplifying enzyme are added to a sample suspected of containing the toxin. The mixture is then cycled thermally to exponentially amplify any of these unique nucleic acid sequences present in the sample. The amplified sequences can be detected by various means, including fluorescence. Detection of the amplified sequences is indicative of the presence of toxin in the original sample. By using more than one set of labeled primers, the method can be used to simultaneously detect several toxins in a sample.

  4. Ambient formic acid in southern California air: A comparison of two methods, Fourier transform infrared spectroscopy and alkaline trap-liquid chromatography with UV detection

    SciTech Connect

    Grosjean, D. ); Tuazon, E.C. ); Fujita, E. )

    1990-01-01

    Formic acid is an ubiquitous component of urban smog. Sources of formic acid in urban air include direct emissions from vehicles and in situ reaction of ozone with olefins. Ambient levels of formic acid in southern California air were first measured some 15 years ago by Hanst et al. using long-path Fourier transform infrared spectroscopy (FTIR). All subsequent studies of formic acid in the Los Angeles area have involved the use of two methods, either FTIR or collection on alkaline traps followed by gas chromatography, ion chromatography, or liquid chromatography analysis with UV detection, ATLC-UV. The Carbon Species Methods Comparison Study (CSMCS), a multilaboratory air quality study carried out in August 1986 at a southern California smog receptor site, provided an opportunity for direct field comparison of the FTIR and alkaline trap methods. The results of the comparison are presented in this brief report.

  5. Solid Phase Micro-extraction (SPME) with In Situ Transesterification: An Easy Method for the Detection of Non-volatile Fatty Acid Derivatives on the Insect Cuticle.

    PubMed

    Kühbandner, Stephan; Ruther, Joachim

    2015-06-01

    Triacylglycerides (TAGs) and other non-volatile fatty acid derivatives (NFADs) occur in large amounts in the internal tissues of insects, but their presence on the insect cuticle is controversially discussed. Most studies investigating cuticular lipids of insects involve solvent extraction, which implies the risk of extracting lipids from internal tissues. Here, we present a new method that overcomes this problem. The method employs solid phase micro-extraction (SPME) to sample NFADs by rubbing the SPME fiber over the insect cuticle. Subsequently, the sampled NFADs are transesterified in situ with trimethyl sulfonium hydroxide (TMSH) into more volatile fatty acid methyl esters (FAMEs), which can be analyzed by standard GC/MS. We performed two types of control experiments to enable significant conclusions: (1) to rule out contamination of the GC/MS system with NFADs, and (2) to exclude the presence of free fatty acids on the insect cuticle, which would also furnish FAMEs after TMSH treatment, and thus might simulate the presence of NFADs. In combination with these two essential control experiments, the described SPME technique can be used to detect TAGs and/or other NFADs on the insect cuticle. We analyzed six insect species from four insect orders with our method and compared the results with conventional solvent extraction followed by ex situ transesterification. Several fatty acids typically found as constituents of TAGs were detected by the SPME method on the cuticle of all species analyzed. A comparison of the two methods revealed differences in the fatty acid compositions of the samples. Saturated fatty acids showed by trend higher relative abundances when sampled with the SPME method, while several minor FAMEs were detected only in the solvent extracts. Our study suggests that TAGs and maybe other NFADs are far more common on the insect cuticle than usually thought.

  6. Limits of detections for the determination of mono- and dicarboxylic acids using gas and liquid chromatographic methods coupled with mass spectrometry

    PubMed Central

    Št’ávová, Jana; Beránek, Josef; Nelson, Eric P.; Diep, Bonnie A.; Kubátová, Alena

    2011-01-01

    The chromatographic separation and instrumental limits of detection (LODs) were obtained for a broad range of C1-C18 monocarboxylic (MCAs) and C2-C14 dicarboxylic acids (DCAs) employing either chemical derivatization followed by gas chromatography-mass spectrometry and flame ionization detection (GC-MS/FID) or direct analysis with liquid chromatography high resolution MS and tandem MS (LC-MS). Suitability, efficiency and stability of reaction products for several derivatization agents used for esterification (BF3/butanol), and trimethysilylation, including trimethylsilyl-N-N-dimethylcarbamate (TMSDMC) and N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) were evaluated. The lowest limits of detection for the majority of compounds below 10 pg (with the exception of acetic acid) were obtained for derivatization with BF3/butanol followed by GC-MS in the total ion current (TIC) mode. Further improvements were achieved when applying either selected ion monitoring (SIM), which decreased the LODs to 1–4 pg or a combination of SIM and TIC (SITI) (2–5 pg). GC-FID provided LODs comparable to those obtained by GC-MS TIC. Both trimethylsilylation (followed by GC-MS) and direct LC-MS/MS analysis yielded LODs of 5– 40 pg for most of the acids. For volatile acids the LODs were higher, e.g., 25 and 590 ng for TMSDMC and BSTFA derivatized formic acid, respectively whereas the LC-MS methods did not allow for the analysis of formic acid at all. PMID:21185238

  7. Sulfenic acid chemistry, detection and cellular lifetime☆

    PubMed Central

    Gupta, Vinayak; Carroll, Kate S.

    2014-01-01

    Background Reactive oxygen species-mediated cysteine sulfenic acid modification has emerged as an important regulatory mechanism in cell signaling. The stability of sulfenic acid in proteins is dictated by the local microenvironment and ability of antioxidants to reduce this modification. Several techniques for detecting this cysteine modification have been developed, including direct and in situ methods. Scope of review This review presents a historical discussion of sulfenic acid chemistry and highlights key examples of this modification in proteins. A comprehensive survey of available detection techniques with advantages and limitations is discussed. Finally, issues pertaining to rates of sulfenic acid formation, reduction, and chemical trapping methods are also covered. Major conclusions Early chemical models of sulfenic acid yielded important insights into the unique reactivity of this species. Subsequent pioneering studies led to the characterization of sulfenic acid formation in proteins. In parallel, the discovery of oxidant-mediated cell signaling pathways and pathological oxidative stress has led to significant interest in methods to detect these modifications. Advanced methods allow for direct chemical trapping of protein sulfenic acids directly in cells and tissues. At the same time, many sulfenic acids are short-lived and the reactivity of current probes must be improved to sample these species, while at the same time, preserving their chemical selectivity. Inhibitors with binding scaffolds can be rationally designed to target sulfenic acid modifications in specific proteins. General significance Ever increasing roles for protein sulfenic acids have been uncovered in physiology and pathology. A more complete understanding of sulfenic acid-mediated regulatory mechanisms will continue to require rigorous and new chemical insights. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and

  8. Rapid and sensitive method for determining free amino acids in honey by gas chromatography with flame ionization or mass spectrometric detection.

    PubMed

    Nozal, Ma J; Bernal, J L; Toribio, M L; Diego, J C; Ruiz, A

    2004-08-20

    This paper describes a rapid, sensitive and specific method for determination of free amino acids in honey involving a new reaction of derivatization and gas chromatography (GC) with flame ionization (FID) and mass spectrometric (MS) detection. The method allows the determination of 22 free amino acids in honey samples in a short time: 8 and 5 min for GC-FID and GC-MS, respectively. Quantitation was performed using Norvaline as internal standard, with detection limits ranging between 0.112 and 1.795 mg/L by GC-FID and between 0.001 and 0.291 mg/L by GC-MS in the selected-ion monitoring mode. The method was validated and applied to a set of 74 honey samples belonging to four different botanical origins: eucaliptus, rosemary, orange and heather. The statistical treatment of data shows a correct classification of different origins over 90%.

  9. Discovery of the antibiotic phosacetamycin via a new mass spectrometry-based method for phosphonic acid detection.

    PubMed

    Evans, Bradley S; Zhao, Changming; Gao, Jiangtao; Evans, Courtney M; Ju, Kou-San; Doroghazi, James R; van der Donk, Wilfred A; Kelleher, Neil L; Metcalf, William W

    2013-05-17

    Naturally occurring phosphonates such as phosphinothricin (Glufosinate, a commercially used herbicide) and fosfomycin (Monurol, a clinically used antibiotic) have proved to be potent and useful biocides. Yet this class of natural products is still an under explored family of secondary metabolites. Discovery of the biosynthetic pathways responsible for the production of these compounds has been simplified by using gene based screening approaches, but detection and identification of the natural products the genes produce have been hampered by a lack of high-throughput methods for screening potential producers under various culture conditions. Here, we present an efficient mass-spectrometric method for the selective detection of natural products containing phosphonate and phosphinate functional groups. We have used this method to identify a new phosphonate metabolite, phosacetamycin, whose structure, biological activity, and biosynthetic gene cluster are reported.

  10. Discovery of the antibiotic phosacetamycin via a new mass spectrometry-based method for phosphonic acid detection

    PubMed Central

    Evans, Bradley S.; Zhao, Changming; Gao, Jiangtao; Evans, Courtney M.; Ju, Kou-San; Doroghazi, James R.; van der Donk, Wilfred A.; Kelleher, Neil L.; Metcalf, William W.

    2013-01-01

    Naturally occurring phosphonates such as phosphinothricin (Glufosinate, a commercially used herbicide) and fosfomycin (Monurol, a clinically used antibiotic) have proved to be potent and useful biocides. Yet this class of natural products is still an under explored family of secondary metabolites. Discovery of the biosynthetic pathways responsible for the production of these compounds has been simplified by using gene based screening approaches, but detection and identification of the natural products the genes produce has been hampered by a lack of high-throughput methods for screening potential producers under various culture conditions. Here we present an efficient mass-spectrometric method for the selective detection of natural products containing phosphonate and phosphinate functional groups. We have used this method to identify a new phosphonate metabolite, phosacetamycin, whose structure, biological activity, and biosynthetic gene cluster are reported. PMID:23474169

  11. Multiplex PCR method for the simultaneous detection of histamine-, tyramine-, and putrescine-producing lactic acid bacteria in foods.

    PubMed

    Marcobal, Angela; de las Rivas, Blanca; Moreno-Arribas, M Victoria; Muñoz, Rosario

    2005-04-01

    In a screening of primers, we have selected three pairs of primers for a multiplex PCR assay for the simultaneous detection of lactic acid bacteria (LAB) strains, which potentially produce histamine, tyramine, and putrescine on fermented foods. These primers were based on sequences from histidine, tyrosine, and ornithine decarboxylases from LAB. Under the optimized conditions, the assay yielded a 367-bp DNA fragment from histidine decarboxylases, a 924-bp fragment from tyrosine decarboxylases, and a 1,446-bp fragment from ornithine decarboxylases. When the DNAs of several target organisms were included in the same reaction, two or three corresponding amplicons of different sizes were observed. This assay was useful for the detection of amine-producing bacteria in control collection strains and in a LAB collection. No amplification was observed with DNA from nonproducing LAB strains. This article is the first describing a multiplex PCR approach for the simultaneous detection of potentially amine-producing LAB in foods. It can be easily incorporated into the routine screening for the accurate selection of starter LAB and in food control laboratories.

  12. A HPLC-fluorescence detection method for determination of phosphatidic acid phosphohydrolase activity: application in human myocardium.

    PubMed

    Burgdorf, Christof; Prey, Antje; Richardt, Gert; Kurz, Thomas

    2008-03-15

    Phosphatidic acid phosphohydrolase (PAP) catalyzes the dephosphorylation of phosphatidic acid (PA) to diacylglycerol, the second messenger responsible for activation of protein kinase C. Despite the crucial role of PAP lipid signaling, there are no data on PAP signaling function in the human heart. Here we present a nonradioactive assay for the investigation of PAP activity in human myocardium using a fluorescent derivative of PA, 2-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)-1-hexadecanoyl-sn-glycero-3-phosphate (BODIPY-PA), as substrate in an in vitro PAP-catalyzed reaction. Unreacted BODIPY-PA was resolved from the PAP products by a binary gradient HPLC system and BODIPY-diacylglycerol was detected by fluorimetry. The reaction proceeded at a linear rate for up to 60 min and increased linearly with increasing amounts of cardiac protein in a range of 0.25 to 8.0 microg. This assay proved to be sensitive for accurate quantitation of total PAP activity, PAP-1 activity, and PAP-2 activity in human atrial tissue and right ventricular endomyocardial biopsies. Total PAP activity was approximately fourfold higher in ventricular myocardium than in atrial tissue. There was negligible PAP-1 activity in atrial myocardium compared with ventricular myocardium, indicating regional differences in activities and distribution pattern of PAP-1 and PAP-2 in the human heart.

  13. Method for isolating nucleic acids

    SciTech Connect

    Hurt, Jr., Richard Ashley; Elias, Dwayne A.

    2015-09-29

    The current disclosure provides methods and kits for isolating nucleic acid from an environmental sample. The current methods and compositions further provide methods for isolating nucleic acids by reducing adsorption of nucleic acids by charged ions and particles within an environmental sample. The methods of the current disclosure provide methods for isolating nucleic acids by releasing adsorbed nucleic acids from charged particles during the nucleic acid isolation process. The current disclosure facilitates the isolation of nucleic acids of sufficient quality and quantity to enable one of ordinary skill in the art to utilize or analyze the isolated nucleic acids for a wide variety of applications including, sequencing or species population analysis.

  14. Simple and Sensitive High-Performance Liquid Chromatography (HPLC) Method with UV Detection for Mycophenolic Acid Assay in Human Plasma. Application to a Bioequivalence Study

    PubMed Central

    Danafar, Hossein; Hamidi, Mehrdad

    2015-01-01

    Purpose: A simple and available reversed-phase high performance liquid chromatography (HPLC) method with UV detection has been developed and validated for mycophenolic acid (MPA) assay in human plasma. Methods: MPA was extracted from plasma with protein precipitation method by acetonitrile: percholeric acid: methanol (75:5:20 v/v/v). The drug separation was achieved using a C8 analytical column and a mobile phase of 0.1M triethylammonium phosphate (pH=5.4)-acetonitril (65:35, v/v), with a flow rate of 1.5 ml/min. The detection wavelength was 304 nm. Limit of detection (LOD) of the method was determined as the lowest MPA concentration producing a signal-to-noise (S/N) ratio of about 3. Limit of quantitation (LOQ) was determined as the lowest MPA concentration capable of being quantitated with enough accuracy and precision. Results: The method showed significant linear response-concentration relationship throughout the MPA concentration range of 0.2-10 µg/ml. A typical linear regression equation of the method was: y = 8.5523 x + 0.094, with x and y representing MPA concentration (in µg/ml) and peak height respectively, and the regression coefficient (r) of 0.9816. The average within-run and between-run variations of 7.81 and 4.78 percent. The average drug recovery from plasma was 95.24 percent throughout the linear concentration range. The limits of detection (LOD) and quantitation (LOQ) of the method were 0.05 and 0.2 µg/ml, respectively. The practical applicability of the method was proven throughout a bioequivalence study. Conclusion: The results showed the acceptable degree of linearity, sensitivity, precision, accuracy and recovery for the method. The method was used successfully for quantitation of MPA in plasma samples of healthy volunteers throughout a bioequivalence study. PMID:26819930

  15. Methods of Melanoma Detection.

    PubMed

    Leachman, Sancy A; Cassidy, Pamela B; Chen, Suephy C; Curiel, Clara; Geller, Alan; Gareau, Daniel; Pellacani, Giovanni; Grichnik, James M; Malvehy, Josep; North, Jeffrey; Jacques, Steven L; Petrie, Tracy; Puig, Susana; Swetter, Susan M; Tofte, Susan; Weinstock, Martin A

    2016-01-01

    Detection and removal of melanoma, before it has metastasized, dramatically improves prognosis and survival. The purpose of this chapter is to (1) summarize current methods of melanoma detection and (2) review state-of-the-art detection methods and technologies that have the potential to reduce melanoma mortality. Current strategies for the detection of melanoma range from population-based educational campaigns and screening to the use of algorithm-driven imaging technologies and performance of assays that identify markers of transformation. This chapter will begin by describing state-of-the-art methods for educating and increasing awareness of at-risk individuals and for performing comprehensive screening examinations. Standard and advanced photographic methods designed to improve reliability and reproducibility of the clinical examination will also be reviewed. Devices that magnify and/or enhance malignant features of individual melanocytic lesions (and algorithms that are available to interpret the results obtained from these devices) will be compared and contrasted. In vivo confocal microscopy and other cellular-level in vivo technologies will be compared to traditional tissue biopsy, and the role of a noninvasive "optical biopsy" in the clinical setting will be discussed. Finally, cellular and molecular methods that have been applied to the diagnosis of melanoma, such as comparative genomic hybridization (CGH), fluorescent in situ hybridization (FISH), and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), will be discussed.

  16. Methods of Endotoxin Detection.

    PubMed

    Su, Wenqiong; Ding, Xianting

    2015-08-01

    Endotoxin, present in the outer membrane of all gram-negative bacteria, can pose serious risks to human health, from irreversible shock to death. Therefore, it is essential to develop sensitive, accurate, and rapid methods for its detection. The rabbit pyrogen test is the first standard technique for endotoxin detection and, nowadays, has been replaced by the Limulus Amoebocyte Lysate test, which is the most popular detection technique for endotoxin. With in-depth understanding of endotoxin, biosensors based on endotoxin-sensing components are promising alternatives to pursue in developing low-cost, easy-operation, and fast-response endotoxin detection techniques. This article summarizes the recent advances of endotoxin detection methods with a particular emphasis on optical and electrochemical biosensors based on various sensing elements ranging from nature biomolecules to artificial materials. As the research and technological revolution continues, the highly integrated and miniaturized commercial devices for sensitively and reliably detecting endotoxin will provide a wide range of applications in people's daily life.

  17. [Clinical usefulness of urine-formed elements' information obtained from bacteria detection by flow cytometry method that uses nucleic acid staining].

    PubMed

    Nakagawa, Hiroko; Yuno, Tomoji; Itho, Kiichi

    2009-03-01

    Recently, specific detection method for Bacteria, by flow cytometry method using nucleic acid staining, was developed as a function of automated urine formed elements analyzer for routine urine testing. Here, we performed a basic study on this bacteria analysis method. In addition, we also have a comparison among urine sediment analysis, urine Gram staining and urine quantitative cultivation, the conventional methods performed up to now. As a result, the bacteria analysis with flow cytometry method that uses nucleic acid staining was excellent in reproducibility, and higher sensitivity compared with microscopic urinary sediment analysis. Based on the ROC curve analysis, which settled urine culture method as standard, cut-off level of 120/microL was defined and its sensitivity = 85.7%, specificity = 88.2%. In the analysis of scattergram, accompanied with urine culture method, among 90% of rod positive samples, 80% of dots were appeared in the area of 30 degrees from axis X. In addition, one case even indicated that analysis of bacteria by flow cytometry and scattergram of time series analysis might be helpful to trace the progress of causative bacteria therefore the information supposed to be clinically significant. Reporting bacteria information with nucleic acid staining flow cytometry method is expected to contribute to a rapid diagnostics and treatment of urinary tract infections. Besides, the contribution to screening examination of microbiology and clinical chemistry, will deliver a more efficient solution to urine analysis.

  18. Comparison and validation of 2 analytical methods for the determination of free fatty acids in dairy products by gas chromatography with flame ionization detection.

    PubMed

    Mannion, David T; Furey, Ambrose; Kilcawley, Kieran N

    2016-07-01

    Accurate quantification of free fatty acids (FFA) in dairy products is important for quality control, nutritional, antimicrobial, authenticity, legislative, and flavor purposes. In this study, the performance of 2 widely used gas chromatographic flame ionization detection methods for determination of FFA in dairy products differing in lipid content and degree of lipolysis were evaluated. We used a direct on-column approach where the isolated FFA extract was injected directly and a derivatization approach where the FFA were esterified in the injector to methyl esters using tetramethylammonium hydroxide as a catalyst. A comprehensive validation was undertaken to establish method linearity, limits of detection, limits of quantification, accuracy, and precision. Linear calibrations of 3 to 700mg/L (R(2)>0.999) and 20 to 700mg/L (R(2)>0.997), and limits of detection and limits of quantification of 0.7 and 3mg/L and 5 and 20mg/L were obtained for the direct injection on-column and the derivatization method, respectively. Intraday precision of 1.5 to 7.2% was obtained for both methods. The direct injection on-column method had the lower levels of limits of detection and quantification, because FFA are directly injected onto the GC as opposed to the split injection used in the derivatization method. However, the direct injection on-column method experienced accumulative column phase deterioration and irreversible FFA absorption because of the acidic nature of the injection extract, which adversely affected method robustness and the quantification of some longer chain FFA. The derivatization method experienced issues with quantification of butyric acid at low concentrations because of coelution with the injection solvent peak, loss of polyunsaturated FFA due to degradation by tetramethylammonium hydroxide, and the periodic emergence of by-product peaks of the tetramethylammonium hydroxide reaction that interfered with the quantification of some short-chain FFA. The

  19. Error detection method

    DOEpatents

    Olson, Eric J.

    2013-06-11

    An apparatus, program product, and method that run an algorithm on a hardware based processor, generate a hardware error as a result of running the algorithm, generate an algorithm output for the algorithm, compare the algorithm output to another output for the algorithm, and detect the hardware error from the comparison. The algorithm is designed to cause the hardware based processor to heat to a degree that increases the likelihood of hardware errors to manifest, and the hardware error is observable in the algorithm output. As such, electronic components may be sufficiently heated and/or sufficiently stressed to create better conditions for generating hardware errors, and the output of the algorithm may be compared at the end of the run to detect a hardware error that occurred anywhere during the run that may otherwise not be detected by traditional methodologies (e.g., due to cooling, insufficient heat and/or stress, etc.).

  20. Method for detecting biomolecules

    DOEpatents

    Huo, Qisheng; Liu, Jun

    2008-08-12

    A method for detecting and measuring the concentration of biomolecules in solution, utilizing a conducting electrode in contact with a solution containing target biomolecules, with a film with controllable pore size distribution characteristics applied to at least one surface of the conducting electrode. The film is functionalized with probe molecules that chemically interact with the target biomolecules at the film surface, blocking indicator molecules present in solution from diffusing from the solution to the electrode, thereby changing the electrochemical response of the electrode

  1. [Viral safety of biologicals: evaluation of hepatitis C virus (HCV) nucleic acid amplification test (NAT) assay and development of concentration method of HCV for sensitive detection by NAT].

    PubMed

    Uchida, Eriko; Yamaguchi, Teruhide

    2010-02-01

    The most important issue for the safety of biological products and blood products derived from human sources is how to prevent transmission of infectious agents. The hepatitis C virus (HCV) is a major public health problem due to its high prevalence. HCV is mainly transmitted by exposure to blood and highly infectious during the early window period with extremely low viral loads. Therefore it is important to develop more sensitive detection methods for HCV. In the case of blood products, both serological test and nucleic acid amplification test (NAT) are required to detect HCV. Since NAT is highly sensitive, establishment of a new standard is required for validation of NAT assay. NAT guideline and establishment of the standard for HCV RNA and HCV genotype panel is introduced in this review. On the other hand, to enhance the sensitivity of virus detection by NAT, a novel viral concentration method using polyethyleneimine (PEI)-conjugated magnetic beads (PEI beads) was developed. PEI beads concentration method is applicable to a wide range of viruses including HCV. Studies using the national standard for HCV RNA, HCV genotype panel and seroconversion panel, suggest that virus concentration method using PEI-beads is useful for improvement of the sensitivity of HCV detection by NAT and applicable to donor screening for HCV.

  2. Detection of KPC Carbapenemase in Pseudomonas aeruginosa Isolated From Clinical Samples Using Modified Hodge Test and Boronic Acid Phenotypic Methods and Their Comparison With the Polymerase Chain Reaction

    PubMed Central

    Falahat, Saeed; Shojapour, Mana; Sadeghi, Abdorrahim

    2016-01-01

    Background Bacterial resistance to antibiotics has become a major source of concern for public health. Pseudomonas aeruginosa strains are important opportunistic pathogens. These bacteria have a high resistance to a wide range of existing antimicrobials and antibiotics. Objectives The present study was performed to evaluate the frequency of KPC in P. aeruginosa isolated from clinical samples of educational hospitals of Arak University of Medical Sciences, using the mentioned phenotypic and genotypic methods. Materials and Methods One hundred and eight non-duplicate clinical isolates of P. aeruginosa were collected from hospitals of Arak University of Medical Sciences, Arak, Iran. Antibacterial susceptibility was determined by the disk diffusion method. KPC production was confirmed by the Modified Hodge Test (MHT), which is a phenotypic test, and combined-disk test with boronic acid and the Polymerase Chain Reaction (PCR). Results In the present study, 13 isolates (12%) of P. aeruginosa were positive for KPC, using PCR. Comparison of the two phenotypic methods used in this study showed that boronic acid is more sensitive than MHT in identification of KPC-producing strains (84.6% vs. 77%). Conclusions Utilization of reliable methods for identifying carbapenemase-producing strains and determining their antibiotic resistance pattern could have a very important role in treatment of infections caused by these strains. A substantial amount of P. aeruginosa isolated from clinical samples of hospitals in Arak (Iran) produce KPC carbapenemase. Due to their low specificity, MHT and boronic acid phenotypic methods could not completely identify KPC-producing P. aeruginosa. However, the sensitivity of boronic acid phenotypic method in detection of KPC was higher than MHT. PMID:27800140

  3. Detection of trace amounts of target DNA from massive background of nucleic acids by using LM-PCR-based pre-amplification method.

    PubMed

    Pan, Xiaoming; Wang, Jing; Zhang, Yanfang; Dong, Ping; Li, Chunchuan; Liang, Xingguo

    2016-11-08

    The sensitivity and specificity of DNA detection may decrease when the target DNA is in very low abundance. To effectively detect trace amounts of target DNA from massive background of nucleic acids, we have developed a powerful multiplex pre-amplification method based on ligation-mediated PCR (LM-PCR) that can greatly enrich multiple target DNAs from massive backgrounds. By employing type IIS restriction endonuclease (REase) and specifically designed oligonucleotide adapters, target DNA can be pre-amplified with high efficiency and sensitivity. Combining with normal PCR, ten copies of target DNA was effectively detected from over 10(8) times more excessive backgrounds with high specificity and ten times more effectively than conventional PCR. In particular, the usage of universal primer in the pre-amplification PCR (pre-amp PCR) ensured that multiple targets could be equivalently amplified, which was confirmed by quantitative PCR (qPCR), indicating it could meet the demands of high-throughput detection. The flexibility and applicability of pre-amp PCR was validated by using different microorganisms DNA as targets and employing two different type IIS REases. The results suggest that the pre-amp PCR method has broad application prospects in various gene detection fields. This article is protected by copyright. All rights reserved.

  4. Simple and rapid analytical method for detection of amino acids in blood using blood spot on filter paper, fast-GC/MS and isotope dilution technique.

    PubMed

    Kawana, Shuichi; Nakagawa, Katsuhiro; Hasegawa, Yuki; Yamaguchi, Seiji

    2010-11-15

    A simple and rapid method for quantitative analysis of amino acids, including valine (Val), leucine (Leu), isoleucine (Ile), methionine (Met) and phenylalanine (Phe), in whole blood has been developed using GC/MS. In this method, whole blood was collected using a filter paper technique, and a 1/8 in. blood spot punch was used for sample preparation. Amino acids were extracted from the sample, and the extracts were purified using cation-exchange resins. The isotope dilution method using ²H₈-Val, ²H₃-Leu, ²H₃-Met and ²H₅-Phe as internal standards was applied. Following propyl chloroformate derivatization, the derivatives were analyzed using fast-GC/MS. The extraction recoveries using these techniques ranged from 69.8% to 87.9%, and analysis time for each sample was approximately 26 min. Calibration curves at concentrations from 0.0 to 1666.7 μmol/l for Val, Leu, Ile and Phe and from 0.0 to 333.3 μmol/l for Met showed good linearity with regression coefficients=1. The method detection limits for Val, Leu, Ile, Met and Phe were 24.2, 16.7, 8.7, 1.5 and 12.9 μmol/l, respectively. This method was applied to blood spot samples obtained from patients with phenylketonuria (PKU), maple syrup urine disease (MSUD), hypermethionine and neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), and the analysis results showed that the concentrations of amino acids that characterize these diseases were increased. These results indicate that this method provides a simple and rapid procedure for precise determination of amino acids in whole blood.

  5. Studies in lipid histochemistry. XIII. The OPA (osmiumtetroxide-periodic acid-alpha-naphthylamine) method for the detection of apolar lipids.

    PubMed

    Elleder, M

    1975-09-29

    A new procedure for the detection of apolar lipids is described. It is a modification of the OTAN method (Adams, 1959) using periodic acid which oxidatively removes lower osmium derivatives from polar sites only, leaving those in apolar lipids intact and demonstrable with alpha-naphthylamine. Control steps for the exclusion of the possible interference of some less polar complex lipids and of lipopigments are described. The described technic is superior to the conventionally used sudan dyes due partly to the fact that only aqueous solutions are employed thus excluding any extraction of lipids, partly to the more distinct coloration.

  6. RAPID AND SIMPLIFIED HPLC METHOD WITH UV DETECTION, PH CONTROL AND SELECTIVE DECHLORINATOR FOR CYANURIC ACID ANALYSIS IN WATER

    EPA Science Inventory

    Cyanuric acid (CA) and chloroisocyanurates are commonly used as standard ingredients in formulations for household bleaches, industrial cleansers, dishwasher compounds, general sanitizers, and chlorine stabilizers. They are very well known for preventing the photolytic decomposi...

  7. Merging a sensitive capillary electrophoresis-ultraviolet detection method with chemometric exploratory data analysis for the determination of phenolic acids and subsequent characterization of avocado fruit.

    PubMed

    Hurtado-Fernández, Elena; Contreras-Gutiérrez, Paulina K; Cuadros-Rodríguez, Luis; Carrasco-Pancorbo, Alegría; Fernández-Gutiérrez, Alberto

    2013-12-15

    Herein we present the development of a powerful CE-UV method able to detect and quantify an important number of phenolic acids in 13 varieties of avocado fruits at 2 ripening stages. All the variables involved in CE separation were exhaustively optimized and the best results were obtained with a capillary of 50 μm i.d. × 50 cm effective length, sodium tetraborate 40 mM at a pH of 9.4, 30 kV, 25 °C, 10s of hydrodynamic injection (0.5 psi) and UV detection at 254 nm. This optimal methodology was fully validated and then applied to different avocado samples. The number of phenolic acids determined varied from 8 to 14 compounds; in general, they were in concentrations ranging from 0.13 ppm to 3.82 ppm, except p-coumaric, benzoic and protocatechuic acids, which were found at higher concentrations. Principal component analysis (PCA) was applied to highlight the differences between varieties and ripening degrees, looking for the most influential analytes.

  8. Applications of a Novel Nucleic Acid Detection Method in Breast Cancer: Analysis of Overexpression of HER-2/neu and FAK

    DTIC Science & Technology

    2002-07-01

    arrays. The ability of nucleic acids to store and transfer information / through Watson- Crick base pairing has intriguing parallels with H ,OOCH3...York, 1988. (54) Glass, A E. G.; Davi". G.; Francis , G. D.; Hill, H. A. 0.; Aston, W. J.; Higgins, L J.; Plotkln, E. V.; Scott, L D. L; Turner, A. P. F

  9. Waveguide disturbance detection method

    DOEpatents

    Korneev, Valeri A.; Nihei, Kurt T.; Myer, Larry R.

    2000-01-01

    A method for detection of a disturbance in a waveguide comprising transmitting a wavefield having symmetric and antisymmetric components from a horizontally and/or vertically polarized source and/or pressure source disposed symmetrically with respect to the longitudinal central axis of the waveguide at one end of the waveguide, recording the horizontal and/or vertical component or a pressure of the wavefield with a vertical array of receivers disposed at the opposite end of the waveguide, separating the wavenumber transform of the wavefield into the symmetric and antisymmetric components, integrating the symmetric and antisymmetric components over a broad frequency range, and comparing the magnitude of the symmetric components and the antisymmetric components to an expected magnitude for the symmetric components and the antisymmetric components for a waveguide of uniform thickness and properties thereby determining whether or not a disturbance is present inside the waveguide.

  10. Satellite detection of volcanic sulphuric acid aerosol

    SciTech Connect

    Baran, A.J.; Foot, J.S.; Dibben, P.C.

    1993-09-03

    This paper presents a new method for detecting sulfuric acid aerosols in the stratosphere. The method is based upon brightness temperature measurements made at two thermal wavelengths. Such measurements can be extracted from polar orbiting satellites. Such clouds are the result of the conversion of sulfur dioxide emissions from large volcanic eruptions, and when formed can have lifetimes of years, and can cause a significant increase in the albedo of the planet. This new method is applied to look at the impact of the Mt Pinatubo eruption on the earths albedo.

  11. Detection of nucleic acids by multiple sequential invasive cleavages

    SciTech Connect

    Hall, Jeff G; Lyamichev, Victor I; Mast, Andrea L; Brow, Mary Ann D

    2012-10-16

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based on charge. The present invention also provides methods for the detection of non-target cleavage products via the formation of a complete and activated protein binding region. The invention further provides sensitive and specific methods for the detection of human cytomegalovirus nucleic acid in a sample.

  12. Detection of nucleic acids by multiple sequential invasive cleavages 02

    DOEpatents

    Hall, Jeff G.; Lyamichev, Victor I.; Mast, Andrea L.; Brow, Mary Ann D.

    2002-01-01

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based on charge. The present invention also provides methods for the detection of non-target cleavage products via the formation of a complete and activated protein binding region. The invention further provides sensitive and specific methods for the detection of human cytomegalovirus nucleic acid in a sample.

  13. Detection of nucleic acids by multiple sequential invasive cleavages

    DOEpatents

    Hall, Jeff G.; Lyamichev, Victor I.; Mast, Andrea L.; Brow, Mary Ann D.

    1999-01-01

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based on charge. The present invention also provides methods for the detection of non-target cleavage products via the formation of a complete and activated protein binding region. The invention further provides sensitive and specific methods for the detection of human cytomegalovirus nucleic acid in a sample.

  14. The Development of a Specific and Sensitive LC-MS-Based Method for the Detection and Quantification of Hydroperoxy- and Hydroxydocosahexaenoic Acids as a Tool for Lipidomic Analysis

    PubMed Central

    Derogis, Priscilla B. M. C.; Freitas, Florêncio P.; Marques, Anna S. F.; Cunha, Daniela; Appolinário, Patricia P.; de Paula, Fernando; Lourenço, Tiago C.; Murgu, Michael; Di Mascio, Paolo; Medeiros, Marisa H. G.; Miyamoto, Sayuri

    2013-01-01

    Docosahexaenoic acid (DHA) is an n-3 polyunsaturated fatty acid that is highly enriched in the brain, and the oxidation products of DHA are present or increased during neurodegenerative disease progression. The characterization of the oxidation products of DHA is critical to understanding the roles that these products play in the development of such diseases. In this study, we developed a sensitive and specific analytical tool for the detection and quantification of twelve major DHA hydroperoxide (HpDoHE) and hydroxide (HDoHE) isomers (isomers at positions 4, 5, 7, 8, 10, 11, 13, 14, 16, 17, 19 and 20) in biological systems. In this study, HpDoHE were synthesized by photooxidation, and the corresponding hydroxides were obtained by reduction with NaBH4. The isolated isomers were characterized by LC-MS/MS, and unique and specific fragment ions were chosen to construct a selected reaction monitoring (SRM) method for the targeted quantitative analysis of each HpDoHE and HDoHE isomer. The detection limits for the LC-MS/MS-SRM assay were 1−670 pg for HpDoHE and 0.5−8.5 pg for HDoHE injected onto a column. Using this method, it was possible to detect the basal levels of HDoHE isomers in both rat plasma and brain samples. Therefore, the developed LC-MS/MS-SRM can be used as an important tool to identify and quantify the hydro(pero)xy derivatives of DHA in biological system and may be helpful for the oxidative lipidomic studies. PMID:24204871

  15. A simple method for the determination of glyphosate and aminomethylphosphonic acid in seawater matrix with high performance liquid chromatography and fluorescence detection.

    PubMed

    Wang, Shu; Liu, Baomin; Yuan, Dongxing; Ma, Jian

    2016-12-01

    Glyphosate (GLYP) is an important herbicide which is also used as the phosphorus source for marine organisms. The wide applications of GLYP can lead to its accumulation in oceans and coastal waters, thus creating environmental issues. However, there is limited methods for detection of GLYP and its degradation product, aminomethylphosphonic acid (AMPA) in saline samples. Therefore, a simple and fast method for the quantification of GLYP and AMPA in seawater matrix has been developed based on the derivatization with 9-fluorenylmethylchloroformate (FMOC-Cl), separation with high performance liquid chromatography (HPLC) and detection with fluorescence detector (FLD). In order to maximize sensitivity, the derivatization procedure was carefully optimized regarding concentration of FMOC-Cl, volume of borate buffer, pH of borate buffer, mixing and derivatization time. The derivatization reaction could be completed within 30min in seawater samples without any additional clean-up or desalting steps. Under the optimized conditions, the developed HPLC method showed a wide linear response (up to several mg/L, R(2)>0.99). The limits of detection were 0.60μg/L and 0.30μg/L for GLYP and AMPA in seawater matrix, respectively. The relative standard deviation was 14.0% for GLYP (1.00mg/L) and 3.1% for AMPA (100μg/L) in saline samples with three different operators (n=24). This method was applied to determine the concentration of GLYP and AMPA in seawater culture media and the recovery data indicated minimal matrix interference. Due to its simplicity, high reproducibility and successful application in seawater culture media analysis, this method is a potentially useful analytical technique for both marine research and environmental science.

  16. Exhaled breath condensate appears to be an unsuitable specimen type for the detection of influenza viruses with nucleic acid-based methods

    PubMed Central

    St. George, Kirsten; Fuschino, Meghan E.; Mokhiber, Katharine; Triner, Wayne; Spivack, Simon D.

    2013-01-01

    Exhaled breath condensate is an airway-derived specimen type that has shown significant promise in the diagnosis of asthma, cancer, and other disorders. The presence of human genomic DNA in this sample type has been proven, but there have been no reports on its utility for the detection of respiratory pathogens. The suitability of exhaled breath condensate for the detection of influenza virus was investigated, as an indication of its potential as a specimen type for respiratory pathogen discovery work. Matched exhaled condensates and nasopharyngeal swabs were collected from 18 adult volunteers. Eleven cases were positive for influenza A virus, and one was positive for influenza B virus. All swab samples tested positive in real-time amplification assays, but only one exhaled condensate, an influenza A positive sample with a very high viral load, tested positive in the real-time RT-PCR assay. Most of the positive nasopharyngeal swab samples inoculated for virus culture also tested positive, whereas influenza virus was not grown from any of the exhaled condensate specimens. It was concluded that influenza viruses are not readily detectable with culture or nucleic acid-based techniques in this sample type, and that exhaled breath condensate may not be suitable for respiratory pathogen investigations with molecular methods. PMID:19733195

  17. Exhaled breath condensate appears to be an unsuitable specimen type for the detection of influenza viruses with nucleic acid-based methods.

    PubMed

    St George, Kirsten; Fuschino, Meghan E; Mokhiber, Katharine; Triner, Wayne; Spivack, Simon D

    2010-01-01

    Exhaled breath condensate is an airway-derived specimen type that has shown significant promise in the diagnosis of asthma, cancer, and other disorders. The presence of human genomic DNA in this sample type has been proven, but there have been no reports on its utility for the detection of respiratory pathogens. The suitability of exhaled breath condensate for the detection of influenza virus was investigated, as an indication of its potential as a specimen type for respiratory pathogen discovery work. Matched exhaled condensates and nasopharyngeal swabs were collected from 18 adult volunteers. Eleven cases were positive for influenza A virus, and one was positive for influenza B virus. All swab samples tested positive in real-time amplification assays, but only one exhaled condensate, an influenza A positive sample with a very high viral load, tested positive in the real-time RT-PCR assay. Most of the positive nasopharyngeal swab samples inoculated for virus culture also tested positive, whereas influenza virus was not grown from any of the exhaled condensate specimens. It was concluded that influenza viruses are not readily detectable with culture or nucleic acid-based techniques in this sample type, and that exhaled breath condensate may not be suitable for respiratory pathogen investigations with molecular methods.

  18. Ultrastructural detection of nucleic acids within heat shock-induced perichromatin granules of HeLa cells by cytochemical and immunocytological methods.

    PubMed

    Charlier, Christine; Lamaye, Françoise; Thelen, Nicolas; Thiry, Marc

    2009-06-01

    The perichromatin granules (PGs) are enigmatic structures of the cell nucleus. The major drawbacks for a biological study are their rare occurrence and their small size in normal conditions. As heat shock has been shown to increase their number, we applied a hyperthermal shock on HeLa cells to investigate the nucleic acid content of PGs by means of cytochemical and immunocytological approaches. These heat shock-induced PGs (hsiPGs) appeared as clusters organized in the form of honeycomb structures and were always associated with some blocks of condensed chromatin, such as the perinucleolar chromatin shell. A stalk connecting the hsiPG to the chromatin could be observed. For the detection of RNA, we applied an immunocytological method involving two anti-RNA antibodies and quantified the gold labelling obtained. The results clearly revealed that hsiPGs contained RNA. Regarding to the detection of DNA, we used three different methods followed by quantitative analyses. The results seemed to indicate that a small amount of DNA was present in hsiPGs. Together, these findings suggest that hsiPGs might be RNP structures associated with particular regions of DNA.

  19. Detection of nucleic acid sequences by invader-directed cleavage

    DOEpatents

    Brow, Mary Ann D.; Hall, Jeff Steven Grotelueschen; Lyamichev, Victor; Olive, David Michael; Prudent, James Robert

    1999-01-01

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The 5' nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based by charge.

  20. Electrochemical Sensors for Detection of Acetylsalicylic Acid

    PubMed Central

    Supalkova, Veronika; Petrek, Jiri; Havel, Ladislav; Krizkova, Sona; Petrlova, Jitka; Adam, Vojtech; Potesil, David; Babula, Petr; Beklova, Miroslava; Horna, Ales; Kizek, Rene

    2006-01-01

    Acetylsalicylic acid (AcSA), or aspirin, was introduced in the late 1890s and has been used to treat a variety of inflammatory conditions. The aim of this work was to suggest electrochemical sensor for acetylsalicylic detection. Primarily, we utilized square wave voltammetry (SWV) using both carbon paste electrode (CPE) and of graphite pencil electrode (GPE) as working ones to indirect determination of AcSA. The principle of indirect determination of AcSA bases in its hydrolysis on salicylic acid (SA), which is consequently detected. Thus, we optimized both determination of SA and conditions for AcSA hydrolysis and found out that the most suitable frequency, amplitude, step potential and the composition and pH of the supporting electrolyte for the determination of SA was 260 Hz, 50 mV, 10 mV and Britton-Robinson buffer (pH 1.81), respectively. The detection limit (S/N = 3) of the SA was 1.3 ng/ml. After that, we aimed on indirect determination of AcSA by SWV CPE. We tested the influence of pH of Britton-Robinson buffer and temperature on yield of hydrolysis, and found out that 100% hydrolysis of AcSA was reached after 80 minutes at pH 1.81 and 90°C. The method for indirect determination of AcSA has been utilized to analyse pharmaceutical drug. The determined amount of AcSA in the pharmaceutical drug was in good agreement with the declared amounts. Moreover, we used GPE for determination of AcSA in a pharmaceutical drug. Base of the results obtained from stationary electrochemical instrument we used flow injection analysis with electrochemical detection to determine of salicylates (SA, AcSA, thiosalicylic acid, 3,5-dinitrosalicylic acid and 5-sulfosalicylic acid – SuSA). We found out that we are able to determine all of detected salicylates directly without any pre-treatment, hydrolysis and so on at units of femtomoles per injection (5 μl).

  1. Biological methods for marine toxin detection.

    PubMed

    Vilariño, Natalia; Louzao, M Carmen; Vieytes, Mercedes R; Botana, Luis M

    2010-07-01

    The presence of marine toxins in seafood poses a health risk to human consumers which has prompted the regulation of the maximum content of marine toxins in seafood in the legislations of many countries. Most marine toxin groups are detected by animal bioassays worldwide. Although this method has well known ethical and technical drawbacks, it is the official detection method for all regulated phycotoxins except domoic acid. Much effort by the scientific and regulatory communities has been focused on the development of alternative techniques that enable the substitution or reduction of bioassays; some of these have recently been included in the official detection method list. During the last two decades several biological methods including use of biosensors have been adapted for detection of marine toxins. The main advances in marine toxin detection using this kind of technique are reviewed. Biological methods offer interesting possibilities for reduction of the number of biosassays and a very promising future of new developments.

  2. Solvent accessible surface area-based hot-spot detection methods for protein-protein and protein-nucleic acid interfaces.

    PubMed

    Munteanu, Cristian R; Pimenta, António C; Fernandez-Lozano, Carlos; Melo, André; Cordeiro, Maria N D S; Moreira, Irina S

    2015-05-26

    Due to the importance of hot-spots (HS) detection and the efficiency of computational methodologies, several HS detecting approaches have been developed. The current paper presents new models to predict HS for protein-protein and protein-nucleic acid interactions with better statistics compared with the ones currently reported in literature. These models are based on solvent accessible surface area (SASA) and genetic conservation features subjected to simple Bayes networks (protein-protein systems) and a more complex multi-objective genetic algorithm-support vector machine algorithms (protein-nucleic acid systems). The best models for these interactions have been implemented in two free Web tools.

  3. Be an acid rain detective

    SciTech Connect

    Atwill, L.

    1982-07-01

    Acid rain is discussed in a question and answer format. The article is aimed at educating sport fishermen on the subject, and also to encourage them to write their congressmen, senators, and the President about the acid rain problem. The article also announces the availability of an acid rain test kit available through the magazine, ''Sports Afield.'' The kit consists of pH-test paper that turns different shades of pink and blue according to the pH of the water tested. The color of the test paper is then compared to a color chart furnished in the kit and an approximate pH can be determined.

  4. Method for nucleic acid isolation using supercritical fluids

    SciTech Connect

    Nivens, David E.; Applegate, Bruce M.

    1999-01-01

    A method for detecting the presence of a microorganism in an environmental sample involves contacting the sample with a supercritical fluid to isolate nucleic acid from the microorganism, then detecting the presence of a particular sequence within the isolated nucleic acid. The nucleic acid may optionally be subjected to further purification.

  5. Method for nucleic acid isolation using supercritical fluids

    DOEpatents

    Nivens, D.E.; Applegate, B.M.

    1999-07-13

    A method is disclosed for detecting the presence of a microorganism in an environmental sample involves contacting the sample with a supercritical fluid to isolate nucleic acid from the microorganism, then detecting the presence of a particular sequence within the isolated nucleic acid. The nucleic acid may optionally be subjected to further purification. 4 figs.

  6. Methods of DNA methylation detection

    NASA Technical Reports Server (NTRS)

    Maki, Wusi Chen (Inventor); Filanoski, Brian John (Inventor); Mishra, Nirankar (Inventor); Rastogi, Shiva (Inventor)

    2010-01-01

    The present invention provides for methods of DNA methylation detection. The present invention provides for methods of generating and detecting specific electronic signals that report the methylation status of targeted DNA molecules in biological samples.Two methods are described, direct and indirect detection of methylated DNA molecules in a nano transistor based device. In the direct detection, methylated target DNA molecules are captured on the sensing surface resulting in changes in the electrical properties of a nano transistor. These changes generate detectable electronic signals. In the indirect detection, antibody-DNA conjugates are used to identify methylated DNA molecules. RNA signal molecules are generated through an in vitro transcription process. These RNA molecules are captured on the sensing surface change the electrical properties of nano transistor thereby generating detectable electronic signals.

  7. Bacillus Spore Inactivation Methods Affect Detection Assays

    PubMed Central

    Dang, Jessica L.; Heroux, Karen; Kearney, John; Arasteh, Ameneh; Gostomski, Mark; Emanuel, Peter A.

    2001-01-01

    Detection of biological weapons is a primary concern in force protection, treaty verification, and safeguarding civilian populations against domestic terrorism. One great concern is the detection of Bacillus anthracis, the causative agent of anthrax. Assays for detection in the laboratory often employ inactivated preparations of spores or nonpathogenic simulants. This study uses several common biodetection platforms to detect B. anthracis spores that have been inactivated by two methods and compares those data to detection of spores that have not been inactivated. The data demonstrate that inactivation methods can affect the sensitivity of nucleic acid- and antibody-based assays for the detection of B. anthracis spores. These effects should be taken into consideration when comparing laboratory results to data collected and assayed during field deployment. PMID:11472945

  8. Method for identifying and quantifying nucleic acid sequence aberrations

    DOEpatents

    Lucas, J.N.; Straume, T.; Bogen, K.T.

    1998-07-21

    A method is disclosed for detecting nucleic acid sequence aberrations by detecting nucleic acid sequences having both a first and a second nucleic acid sequence type, the presence of the first and second sequence type on the same nucleic acid sequence indicating the presence of a nucleic acid sequence aberration. The method uses a first hybridization probe which includes a nucleic acid sequence that is complementary to a first sequence type and a first complexing agent capable of attaching to a second complexing agent and a second hybridization probe which includes a nucleic acid sequence that selectively hybridizes to the second nucleic acid sequence type over the first sequence type and includes a detectable marker for detecting the second hybridization probe. 11 figs.

  9. Method for identifying and quantifying nucleic acid sequence aberrations

    DOEpatents

    Lucas, Joe N.; Straume, Tore; Bogen, Kenneth T.

    1998-01-01

    A method for detecting nucleic acid sequence aberrations by detecting nucleic acid sequences having both a first and a second nucleic acid sequence type, the presence of the first and second sequence type on the same nucleic acid sequence indicating the presence of a nucleic acid sequence aberration. The method uses a first hybridization probe which includes a nucleic acid sequence that is complementary to a first sequence type and a first complexing agent capable of attaching to a second complexing agent and a second hybridization probe which includes a nucleic acid sequence that selectively hybridizes to the second nucleic acid sequence type over the first sequence type and includes a detectable marker for detecting the second hybridization probe.

  10. Double injection/single detection asymmetric flow injection manifold for spectrophotometric determination of ascorbic acid and uric acid: Selection the optimal conditions by MCDM approach based on different criteria weighting methods

    NASA Astrophysics Data System (ADS)

    Boroumand, Samira; Chamjangali, Mansour Arab; Bagherian, Ghadamali

    2017-03-01

    A simple and sensitive double injection/single detector flow injection analysis (FIA) method is proposed for the simultaneous kinetic determination of ascorbic acid (AA) and uric acid (UA). This method is based upon the difference between the rates of the AA and UA reactions with Fe3 + in the presence of 1, 10-phenanthroline (phen). The absorbance of Fe2 +/1, 10-phenanthroline (Fe-phen) complex obtained as the product was measured spectrophotometrically at 510 nm. To reach a good accuracy in the differential kinetic determination via the mathematical manipulations of the transient signals, different criteria were considered in the selection of the optimum conditions. The multi criteria decision making (MCDM) approach was applied for the selection of the optimum conditions. The importance weights of the evaluation criteria were determined using the analytic hierarchy process, entropy method, and compromised weighting (CW). The experimental conditions (alternatives) were ranked by the technique for order preference by similarity to an ideal solution. Under the selected optimum conditions, the obtained analytical signals were linear in the ranges of 0.50-5.00 and 0.50-4.00 mg L- 1 for AA and UA, respectively. The 3σ detection limits were 0.07 mg L- 1 for AA and 0.12 mg L- 1 for UA. The relative standard deviations for four replicate determinations of AA and UA were 2.03% and 3.30% respectively. The method was also applied for the analysis of analytes in the blood serum, Vitamine C tablets, and tap water with satisfactory results.

  11. Double injection/single detection asymmetric flow injection manifold for spectrophotometric determination of ascorbic acid and uric acid: Selection the optimal conditions by MCDM approach based on different criteria weighting methods.

    PubMed

    Boroumand, Samira; Chamjangali, Mansour Arab; Bagherian, Ghadamali

    2017-03-05

    A simple and sensitive double injection/single detector flow injection analysis (FIA) method is proposed for the simultaneous kinetic determination of ascorbic acid (AA) and uric acid (UA). This method is based upon the difference between the rates of the AA and UA reactions with Fe(3+) in the presence of 1, 10-phenanthroline (phen). The absorbance of Fe(2+)/1, 10-phenanthroline (Fe-phen) complex obtained as the product was measured spectrophotometrically at 510nm. To reach a good accuracy in the differential kinetic determination via the mathematical manipulations of the transient signals, different criteria were considered in the selection of the optimum conditions. The multi criteria decision making (MCDM) approach was applied for the selection of the optimum conditions. The importance weights of the evaluation criteria were determined using the analytic hierarchy process, entropy method, and compromised weighting (CW). The experimental conditions (alternatives) were ranked by the technique for order preference by similarity to an ideal solution. Under the selected optimum conditions, the obtained analytical signals were linear in the ranges of 0.50-5.00 and 0.50-4.00mgL(-1) for AA and UA, respectively. The 3σ detection limits were 0.07mgL(-1) for AA and 0.12mgL(-1) for UA. The relative standard deviations for four replicate determinations of AA and UA were 2.03% and 3.30% respectively. The method was also applied for the analysis of analytes in the blood serum, Vitamine C tablets, and tap water with satisfactory results.

  12. GMDD: a database of GMO detection methods

    PubMed Central

    Dong, Wei; Yang, Litao; Shen, Kailin; Kim, Banghyun; Kleter, Gijs A; Marvin, Hans JP; Guo, Rong; Liang, Wanqi; Zhang, Dabing

    2008-01-01

    Background Since more than one hundred events of genetically modified organisms (GMOs) have been developed and approved for commercialization in global area, the GMO analysis methods are essential for the enforcement of GMO labelling regulations. Protein and nucleic acid-based detection techniques have been developed and utilized for GMOs identification and quantification. However, the information for harmonization and standardization of GMO analysis methods at global level is needed. Results GMO Detection method Database (GMDD) has collected almost all the previous developed and reported GMOs detection methods, which have been grouped by different strategies (screen-, gene-, construct-, and event-specific), and also provide a user-friendly search service of the detection methods by GMO event name, exogenous gene, or protein information, etc. In this database, users can obtain the sequences of exogenous integration, which will facilitate PCR primers and probes design. Also the information on endogenous genes, certified reference materials, reference molecules, and the validation status of developed methods is included in this database. Furthermore, registered users can also submit new detection methods and sequences to this database, and the newly submitted information will be released soon after being checked. Conclusion GMDD contains comprehensive information of GMO detection methods. The database will make the GMOs analysis much easier. PMID:18522755

  13. Multiplexed detection of fungal nucleic acid signatures.

    PubMed

    Diaz, Mara R; Dunbar, Sherry A; Jacobson, James W

    2008-04-01

    Diagnoses of opportunistic mycotic infections constitute an increasing clinical problem. Conventional diagnostic tests are time consuming and lack specificity and sensitivity for accurate and timely prognoses. This unit provides a comprehensive description of a fungal detection method that combines nucleic acid signatures with flow cytometry. The multiplexed assay, which uses xMAP technology, consists of unique fluorescent microspheres covalently bound to species-specific fungal oligonucleotide probes. In the presence of the complementary target sequence, the probe hybridizes to its biotinylated target. Quantification of the reaction is based on the fluorescence signal of the reporter molecule that binds to the biotin moieties of the target. The assay can be expanded to include other microorganisms and has the capability to simultaneously test 100 different fungal probes per tube/well. The speed, flexibility in design, and high-throughput capability makes this assay an attractive diagnostic tool for fungal infections and other related maladies.

  14. Method for detecting an element

    DOEpatents

    Blackwood, Larry G.; Reber, Edward L.; Rohde, Kenneth W.

    2007-02-06

    A method for detecting an element is disclosed and which includes the steps of providing a gamma-ray spectrum which depicts, at least in part, a test region having boundaries, and which has a small amount of the element to be detected; providing a calculation which detects the small amount of the element to be detected; and providing a moving window and performing the calculation within the moving window, and over a range of possible window boundaries within the test region to determine the location of the optimal test region within the gamma-ray spectrum.

  15. Reliability of nucleic acid amplification methods for detection of Chlamydia trachomatis in urine: results of the first international collaborative quality control study among 96 laboratories.

    PubMed

    Verkooyen, Roel P; Noordhoek, Gerda T; Klapper, Paul E; Reid, Jim; Schirm, Jurjen; Cleator, Graham M; Ieven, Margareta; Hoddevik, Gunnar

    2003-07-01

    The first European Quality Control Concerted Action study was organized to assess the ability of laboratories to detect Chlamydia trachomatis in a panel of urine samples by nucleic acid amplification tests (NATs). The panel consisted of lyophilized urine samples, including three negative, two strongly positive, and five weakly positive samples. Ninety-six laboratories in 22 countries participated with a total of 102 data sets. Of 204 strongly positive samples 199 (97.5%) were correctly reported, and of 506 weakly positive samples 466 (92.1%) were correctly reported. In 74 (72.5%) data sets correct results were reported on all samples, and 17 data sets (16.7%) showed either one false-negative or one false-positive result. In another 11 data sets, two or more incorrect results were reported, and two data sets reported a false-positive result on one negative sample. The Roche COBAS Amplicor test was performed in 44 (43%) data sets, the Abbott LCx assay was performed in 31 (30%) data sets, the Roche Amplicor manual assay was performed in 9 (9%) data sets, an in-house PCR was performed in 9 (9%) data sets, the Becton Dickinson ProbeTec ET assay was performed in 5 (4.9%) data sets, and the GenProbe TMA assay was performed in 4 (3.9%) data sets. The results of the Roche Amplicor manual (95.6% correct), COBAS Amplicor (97.0%), and Abbott LCx (94.8%) tests were comparable (P = 0.48). The results with the in-house PCR, BD ProbeTec ET, and GenProbe TMA tests were reported correctly in 88.6, 98, and 92.5% of the tests, respectively. Freeze-drying of clinical urine specimens proved to be a successful method for generating standardized, stable, and easy-to-transport samples for the detection of C. trachomatis by using NATs. Although the results, especially the specificity, for this proficiency panel were better than most quality control studies, sensitivity problems occurred frequently, underlining the need for good laboratory practice and reference reagents to monitor the

  16. Outlier detection method in GEEs.

    PubMed

    Pardo, María Del Carmen; Hobza, Tomáš

    2014-09-01

    The generalized estimating equations (GEEs) method has become quite useful in modeling correlated data. However, diagnostic tools to check that the selected final model fits the data as accurately as possible have not been explored intensively. In this paper, an outlier detection technique is developed based on the use of the "working" score test statistic to test an appropriate mean-shift model in the context of longitudinal studies based on GEEs. Through a simulation study it has been shown that this method correctly singled out the outlier when the data set had a known outlier. The method is applied to a set of data to illustrate the outlier detection procedure in GEEs.

  17. Automated Methods for Multiplexed Pathogen Detection

    SciTech Connect

    Straub, Tim M.; Dockendorff, Brian P.; Quinonez-Diaz, Maria D.; Valdez, Catherine O.; Shutthanandan, Janani I.; Tarasevich, Barbara J.; Grate, Jay W.; Bruckner-Lea, Cindy J.

    2005-09-01

    Detection of pathogenic microorganisms in environmental samples is a difficult process. Concentration of the organisms of interest also co-concentrates inhibitors of many end-point detection methods, notably, nucleic acid methods. In addition, sensitive, highly multiplexed pathogen detection continues to be problematic. The primary function of the BEADS (Biodetection Enabling Analyte Delivery System) platform is the automated concentration and purification of target analytes from interfering substances, often present in these samples, via a renewable surface column. In one version of BEADS, automated immunomagnetic separation (IMS) is used to separate cells from their samples. Captured cells are transferred to a flow-through thermal cycler where PCR, using labeled primers, is performed. PCR products are then detected by hybridization to a DNA suspension array. In another version of BEADS, cell lysis is performed, and community RNA is purified and directly labeled. Multiplexed detection is accomplished by direct hybridization of the RNA to a planar microarray. The integrated IMS/PCR version of BEADS can successfully purify and amplify 10 E. coli O157:H7 cells from river water samples. Multiplexed PCR assays for the simultaneous detection of E. coli O157:H7, Salmonella, and Shigella on bead suspension arrays was demonstrated for the detection of as few as 100 cells for each organism. Results for the RNA version of BEADS are also showing promising results. Automation yields highly purified RNA, suitable for multiplexed detection on microarrays, with microarray detection specificity equivalent to PCR. Both versions of the BEADS platform show great promise for automated pathogen detection from environmental samples. Highly multiplexed pathogen detection using PCR continues to be problematic, but may be required for trace detection in large volume samples. The RNA approach solves the issues of highly multiplexed PCR and provides ''live vs. dead'' capabilities. However

  18. Automated methods for multiplexed pathogen detection.

    PubMed

    Straub, Timothy M; Dockendorff, Brian P; Quiñonez-Díaz, Maria D; Valdez, Catherine O; Shutthanandan, Janani I; Tarasevich, Barbara J; Grate, Jay W; Bruckner-Lea, Cynthia J

    2005-09-01

    Detection of pathogenic microorganisms in environmental samples is a difficult process. Concentration of the organisms of interest also co-concentrates inhibitors of many end-point detection methods, notably, nucleic acid methods. In addition, sensitive, highly multiplexed pathogen detection continues to be problematic. The primary function of the BEADS (Biodetection Enabling Analyte Delivery System) platform is the automated concentration and purification of target analytes from interfering substances, often present in these samples, via a renewable surface column. In one version of BEADS, automated immunomagnetic separation (IMS) is used to separate cells from their samples. Captured cells are transferred to a flow-through thermal cycler where PCR, using labeled primers, is performed. PCR products are then detected by hybridization to a DNA suspension array. In another version of BEADS, cell lysis is performed, and community RNA is purified and directly labeled. Multiplexed detection is accomplished by direct hybridization of the RNA to a planar microarray. The integrated IMS/PCR version of BEADS can successfully purify and amplify 10 E. coli O157:H7 cells from river water samples. Multiplexed PCR assays for the simultaneous detection of E. coli O157:H7, Salmonella, and Shigella on bead suspension arrays was demonstrated for the detection of as few as 100 cells for each organism. Results for the RNA version of BEADS are also showing promising results. Automation yields highly purified RNA, suitable for multiplexed detection on microarrays, with microarray detection specificity equivalent to PCR. Both versions of the BEADS platform show great promise for automated pathogen detection from environmental samples. Highly multiplexed pathogen detection using PCR continues to be problematic, but may be required for trace detection in large volume samples. The RNA approach solves the issues of highly multiplexed PCR and provides "live vs. dead" capabilities. However

  19. Development and validation of a high-throughput based on liquid chromatography with ultraviolet absorption and mass spectrometry detection method for quantitation of cichoric acid in Echinacea purpurea aerial-based dietary supplements.

    PubMed

    Chen, Pei

    2006-01-01

    A new, rapid, and reproducible reversed-phased liquid chromatography (LC) method with ultraviolet (UV) absorption and/or mass spectrometry (MS) detection has been developed and validated for quantitation of cichoric acid, a major constituent of Echinacea spp. The method involves the use of a short Phenomenex Hydro-RP C18 column (4 microm, 50 mm x 3.0 mm id) and a simple isocratic mobile phase profile. Both UV (diode array detector) and selective-ion monitoring (SIM) at m/z 472.8 were used for quantitation of cichoric acid. The limit of detection was 0.75 ng for UV and 0.15 ng for MS-SIM, and the limit of quantitation was is 2.5 ng for UV and 0.5 ng for MS-SIM. Water-methanol (1 + 1) soluble extracts of 6 commercially available Echinacea purpurea aerial parts-based dietary supplements (EPADS). EPADS were first profiled using a traditional HPLC-UV method. Their UV chromatograms were compared, and cichoric acid was identified to be a key biomarker for EPADS. Then the samples were analyzed by the fast LC-UV/MS method. The turnaround time for a single analysis was 3 min, compared to 15 to 60 min needed for traditional reported LC methods. The high-throughput method was able to separate the cichoric acid peak from peaks of other components in extracts of complex matrixes of EPADS.

  20. Nucleic acid in-situ hybridization detection of infectious agents

    NASA Astrophysics Data System (ADS)

    Thompson, Curtis T.

    2000-04-01

    Limitations of traditional culture methods and newer polymerase chain reaction (PCR)-based methods for detection and speciation of infectious agents demonstrate the need for more rapid and better diagnostics. Nucleic acid hybridization is a detection technology that has gained wide acceptance in cancer and prenatal cytogenetics. Using a modification of the nucleic acid hybridization technique known as fluorescence in-situ hybridization, infectious agents can be detected in a variety of specimens with high sensitivity and specificity. The specimens derive from all types of human and animal sources including body fluids, tissue aspirates and biopsy material. Nucleic acid hybridization can be performed in less than one hour. The result can be interpreted either using traditional fluorescence microscopy or automated platforms such as micro arrays. This paper demonstrates proof of concept for nucleic acid hybridization detection of different infectious agents. Interpretation within a cytologic and histologic context is possible with fluorescence microscopic analysis, thereby providing confirmatory evidence of hybridization. With careful probe selection, nucleic acid hybridization promises to be a highly sensitive and specific practical diagnostic alternative to culture, traditional staining methods, immunohistochemistry and complicated nucleic acid amplification tests.

  1. High-performance liquid chromatographic method for the simultaneous detection of the adulteration of cereal flours with melamine and related triazine by-products ammeline, ammelide, and cyanuric acid.

    PubMed

    Ehling, S; Tefera, S; Ho, I P

    2007-12-01

    Melamine has been used for the adulteration of cereal flours in order to increase their apparent protein content. Crude melamine may contain several by-products, i.e. ammeline, ammelide, and cyanuric acid. The simultaneous analysis of all four chemicals is difficult because of the formation of an insoluble salt between melamine and cyanuric acid. A simple and convenient high-performance liquid chromatography (HPLC) method for the detection of the adulteration of cereal flours with all four chemicals is proposed herein. The precipitate formation between melamine and cyanuric acid was prevented by using alkaline conditions (pH 11-12) for both standards preparation and sample extraction. The method uses matrix-matching, which involves the construction of a calibration curve on a blank (negative control) matrix, which is then used for the quantitation of melamine and by-products in adulterated (positive) samples. Matrix-matching compensates for analyte losses during sample preparation, and for matrix effects. The method was successfully applied to wheat, corn, and rice flours, and is expected to be applicable (with some modifications) to soy flour as well. The method allows for the detection of melamine, ammeline, and ammelide at approximately 5 microg g(-1), and cyanuric acid at approximately 90 microg g(-1) in wheat flour.

  2. A novel method for detection of apoptosis

    SciTech Connect

    Zagariya, Alexander M.

    2012-04-15

    There are two different Angiotensin II (ANG II) peptides in nature: Human type (ANG II) and Bovine type (ANG II*). These eight amino acid peptides differ only at position 5 where Valine is replaced by Isoleucine in the Bovine type. They are present in all species studied so far. These amino acids are different by only one atom of carbon. This difference is so small, that it will allow any of ANG II, Bovine or Human antibodies to interact with all species and create a universal method for apoptosis detection. ANG II concentrations are found at substantially higher levels in apoptotic, compared to non-apoptotic, tissues. ANG II accumulation can lead to DNA damage, mutations, carcinogenesis and cell death. We demonstrate that Bovine antiserum can be used for universal detection of apoptosis. In 2010, the worldwide market for apoptosis detection reached the $20 billion mark and significantly increases each year. Most commercially available methods are related to Annexin V and TUNNEL. Our new method based on ANG II is more widely known to physicians and scientists compared to previously used methods. Our approach offers a novel alternative for assessing apoptosis activity with enhanced sensitivity, at a lower cost and ease of use.

  3. Method for detecting toxic gases

    DOEpatents

    Stetter, J.R.; Zaromb, S.; Findlay, M.W. Jr.

    1991-10-08

    A method is disclosed which is capable of detecting low concentrations of a pollutant or other component in air or other gas. This method utilizes a combination of a heating filament having a catalytic surface of a noble metal for exposure to the gas and producing a derivative chemical product from the component. An electrochemical sensor responds to the derivative chemical product for providing a signal indicative of the product. At concentrations in the order of about 1-100 ppm of tetrachloroethylene, neither the heating filament nor the electrochemical sensor is individually capable of sensing the pollutant. In the combination, the heating filament converts the benzyl chloride to one or more derivative chemical products which may be detected by the electrochemical sensor. 6 figures.

  4. Survey of Anomaly Detection Methods

    SciTech Connect

    Ng, B

    2006-10-12

    This survey defines the problem of anomaly detection and provides an overview of existing methods. The methods are categorized into two general classes: generative and discriminative. A generative approach involves building a model that represents the joint distribution of the input features and the output labels of system behavior (e.g., normal or anomalous) then applies the model to formulate a decision rule for detecting anomalies. On the other hand, a discriminative approach aims directly to find the decision rule, with the smallest error rate, that distinguishes between normal and anomalous behavior. For each approach, we will give an overview of popular techniques and provide references to state-of-the-art applications.

  5. A rapid and simple method for the determination of 3,4-dihydroxyphenylacetic acid, norepinephrine, dopamine, and serotonin in mouse brain homogenate by HPLC with fluorimetric detection.

    PubMed

    De Benedetto, Giuseppe Egidio; Fico, Daniela; Pennetta, Antonio; Malitesta, Cosimino; Nicolardi, Giuseppe; Lofrumento, Dario Domenico; De Nuccio, Francesco; La Pesa, Velia

    2014-09-01

    A fast and simple isocratic high-performance liquid chromatography method for the determination of 3,4-dihydroxyphenylacetic acid (DOPAC), norepinephrine (NE), dopamine (DA), and serotonin (5-HT) in homogenate samples of mouse striatum employing the direct fluorescence of the neurotransmitters is described. The method has been optimized and validated. The analytes were separated in 15min on a reversed-phase column (C18) with acetate buffer (pH 4.0, 12mM)-methanol (86:14, v/v) as mobile phase; the flow rate was 1ml/min. The fluorescence measurements were carried out at 320nm with excitation at 279nm. The calibration curve for DA was linear up to about 2.5μg/ml, with a coefficient of determination (r(2)) of 0.9995 with a lower limit of quantification of 0.031μg/ml. Since the procedure does not involve sample pre-purification or derivatisation, the recovery ranged from 97% to 102% and relative standard deviation (RSD) was better than 2.9%, the use of the internal standard is not mandatory, further simplifying the method. Similar performance was obtained for the other analytes. As a result, thanks to its simplicity, rapidity and adequate working range, the method can be used for the determination of 3,4-dihydroxyphenylacetic acid, dopamine, norepinephrine and serotonin in animal tissues. An experimental 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson-like disease has been used to demonstrate the method is fit-for-purpose.

  6. Development of sample preparation method for isoliquiritigenin, liquiritin, and glycyrrhizic acid analysis in licorice by ionic liquids-ultrasound based extraction and high-performance liquid chromatography detection.

    PubMed

    Yang, Lei; Li, Li-li; Liu, Ting-ting; Zu, Yuan-gang; Yang, Feng-jian; Zhao, Chun-jian; Zhang, Lin; Chen, Xiao-qiang; Zhang, Zhong-hua

    2013-05-01

    An ionic liquid-based ultrasonic-assisted extraction (ILUAE) method had been used for the effective extraction of isoliquiritigenin (IQ), liquiritin (LQ) and glycyrrhizic acid (GA) from licorice. The ionic liquids with different cations and anions were investigated in this work and 0.5 M 1-butyl-3-methylimidazolium bromide solution was selected as solvent. In addition, the technical parameters including soaking time, solid-liquid ratio, ultrasonic power and time were optimized. Compared with the conventional solvent extraction, the proposed approach exhibited higher efficiency, which indicated the ILUAE was an efficient, rapid and simple sample preparation technique. There was no degradation of the target analytes had been observed at the optimum conditions which was evidenced by the stability studies performed with standard of IQ, LQ and GA. The proposed method also showed high reproducibility and was environmental friendly.

  7. Developing nucleic acid-based electrical detection systems

    PubMed Central

    Gabig-Ciminska, Magdalena

    2006-01-01

    Development of nucleic acid-based detection systems is the main focus of many research groups and high technology companies. The enormous work done in this field is particularly due to the broad versatility and variety of these sensing devices. From optical to electrical systems, from label-dependent to label-free approaches, from single to multi-analyte and array formats, this wide range of possibilities makes the research field very diversified and competitive. New challenges and requirements for an ideal detector suitable for nucleic acid analysis include high sensitivity and high specificity protocol that can be completed in a relatively short time offering at the same time low detection limit. Moreover, systems that can be miniaturized and automated present a significant advantage over conventional technology, especially if detection is needed in the field. Electrical system technology for nucleic acid-based detection is an enabling mode for making miniaturized to micro- and nanometer scale bio-monitoring devices via the fusion of modern micro- and nanofabrication technology and molecular biotechnology. The electrical biosensors that rely on the conversion of the Watson-Crick base-pair recognition event into a useful electrical signal are advancing rapidly, and recently are receiving much attention as a valuable tool for microbial pathogen detection. Pathogens may pose a serious threat to humans, animal and plants, thus their detection and analysis is a significant element of public health. Although different conventional methods for detection of pathogenic microorganisms and their toxins exist and are currently being applied, improvements of molecular-based detection methodologies have changed these traditional detection techniques and introduced a new era of rapid, miniaturized and automated electrical chip detection technologies into pathogen identification sector. In this review some developments and current directions in nucleic acid-based electrical

  8. Nucleic acid arrays and methods of synthesis

    DOEpatents

    Sabanayagam, Chandran R.; Sano, Takeshi; Misasi, John; Hatch, Anson; Cantor, Charles

    2001-01-01

    The present invention generally relates to high density nucleic acid arrays and methods of synthesizing nucleic acid sequences on a solid surface. Specifically, the present invention contemplates the use of stabilized nucleic acid primer sequences immobilized on solid surfaces, and circular nucleic acid sequence templates combined with the use of isothermal rolling circle amplification to thereby increase nucleic acid sequence concentrations in a sample or on an array of nucleic acid sequences.

  9. Qualitative detection of avian influenza A (H5N1) viruses: a comparative evaluation of four real-time nucleic acid amplification methods.

    PubMed

    Chantratita, Wasun; Sukasem, Chonlaphat; Kaewpongsri, Supaporn; Srichunrusami, Chutatip; Pairoj, Wantanit; Thitithanyanont, Arunee; Chaichoune, Kridsada; Ratanakron, Parntep; Songserm, Thaweesak; Damrongwatanapokin, Sudarat; Landt, Olfert

    2008-01-01

    The aim of this study was to determine the performance of real-time amplification based methods - NASBA, TaqMan, RT-FRET, and RT-PCR LUXtrade mark formats - for the detection of influenza A (H5N1) virus RNA. In an analysis of 54 samples obtained from a range of animal species in Thailand during the period 2003-2006, results showed that the NASBA (H5=98.2%, N1=96.3%), TaqMan (H5=98.2%, N1=96.3%) and FRET (H5=98.2%, N1=96.3%) had significantly higher rates of positive detection than LUX (H5=94.4%, N1=50.0%; P<0.001) for influenza A, H5 and N1 isolates. There were no false-positive results from any methods used in the negative-control group of samples. The limits of analytical detection were at least 10copies/reaction in real-time NASBA and LUX assays, while FRET and TaqMan assay appeared to be less sensitive at > or =100copies/reaction. The assays were relatively specific without cross-reactivity to a number of other influenza strains or viral pathogens. In conclusion, our study demonstrated that real-time NASBA, TaqMan and FRET assays can be used to detect influenza A (H5N1) from a wide range of hosts, and be specific for H5N1 samples obtained during different outbreaks (2003-2006). All assays provided the benefit of rapid influenza H5N1 identification for early diagnosis, in the range of hours, and they are well suited to high throughput analyses.

  10. Method for detecting toxic gases

    DOEpatents

    Stetter, Joseph R.; Zaromb, Solomon; Findlay, Jr., Melvin W.

    1991-01-01

    A method capable of detecting low concentrations of a pollutant or other component in air or other gas, utilizing a combination of a heating filament having a catalytic surface of a noble metal for exposure to the gas and producing a derivative chemical product from the component, and an electrochemical sensor responsive to the derivative chemical product for providing a signal indicative of the product. At concentrations in the order of about 1-100 ppm of tetrachloroethylene, neither the heating filament nor the electrochemical sensor is individually capable of sensing the pollutant. In the combination, the heating filament converts the benzyl chloride to one or more derivative chemical products which may be detected by the electrochemical sensor.

  11. Bacteria detection instrument and method

    NASA Technical Reports Server (NTRS)

    Renner, W.; Fealey, R. D. (Inventor)

    1972-01-01

    A method and apparatus for screening a sample fluid for bacterial presence are disclosed wherein the fluid sample is mixed with culture media of sufficient quantity to permit bacterial growth in order to obtain a test solution. The concentration of oxygen dissolved in the test solution is then monitored using the potential difference between a reference electrode and a noble metal electrode which are in contact with the test solution. The change in oxygen concentration which occurs during a period of time as indicated by the electrode potential difference is compared with a detection criterion which exceeds the change which would occur absent bacteria.

  12. Efficient, validated method for detection of mycobacterial growth in liquid culture media by use of bead beating, magnetic-particle-based nucleic acid isolation, and quantitative PCR.

    PubMed

    Plain, Karren M; Waldron, Anna M; Begg, Douglas J; de Silva, Kumudika; Purdie, Auriol C; Whittington, Richard J

    2015-04-01

    Pathogenic mycobacteria are difficult to culture, requiring specialized media and a long incubation time, and have complex and exceedingly robust cell walls. Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne's disease, a chronic wasting disease of ruminants, is a typical example. Culture of MAP from the feces and intestinal tissues is a commonly used test for confirmation of infection. Liquid medium offers greater sensitivity than solid medium for detection of MAP; however, support for the BD Bactec 460 system commonly used for this purpose has been discontinued. We previously developed a new liquid culture medium, M7H9C, to replace it, with confirmation of growth reliant on PCR. Here, we report an efficient DNA isolation and quantitative PCR methodology for the specific detection and confirmation of MAP growth in liquid culture media containing egg yolk. The analytical sensitivity was at least 10(4)-fold higher than a commonly used method involving ethanol precipitation of DNA and conventional PCR; this may be partly due to the addition of a bead-beating step to manually disrupt the cell wall of the mycobacteria. The limit of detection, determined using pure cultures of two different MAP strains, was 100 to 1,000 MAP organisms/ml. The diagnostic accuracy was confirmed using a panel of cattle fecal (n=54) and sheep fecal and tissue (n=90) culture samples. This technique is directly relevant for diagnostic laboratories that perform MAP cultures but may also be applicable to the detection of other species, including M. avium and M. tuberculosis.

  13. Efficient, Validated Method for Detection of Mycobacterial Growth in Liquid Culture Media by Use of Bead Beating, Magnetic-Particle-Based Nucleic Acid Isolation, and Quantitative PCR

    PubMed Central

    Waldron, Anna M.; Begg, Douglas J.; de Silva, Kumudika; Purdie, Auriol C.; Whittington, Richard J.

    2015-01-01

    Pathogenic mycobacteria are difficult to culture, requiring specialized media and a long incubation time, and have complex and exceedingly robust cell walls. Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne's disease, a chronic wasting disease of ruminants, is a typical example. Culture of MAP from the feces and intestinal tissues is a commonly used test for confirmation of infection. Liquid medium offers greater sensitivity than solid medium for detection of MAP; however, support for the BD Bactec 460 system commonly used for this purpose has been discontinued. We previously developed a new liquid culture medium, M7H9C, to replace it, with confirmation of growth reliant on PCR. Here, we report an efficient DNA isolation and quantitative PCR methodology for the specific detection and confirmation of MAP growth in liquid culture media containing egg yolk. The analytical sensitivity was at least 104-fold higher than a commonly used method involving ethanol precipitation of DNA and conventional PCR; this may be partly due to the addition of a bead-beating step to manually disrupt the cell wall of the mycobacteria. The limit of detection, determined using pure cultures of two different MAP strains, was 100 to 1,000 MAP organisms/ml. The diagnostic accuracy was confirmed using a panel of cattle fecal (n = 54) and sheep fecal and tissue (n = 90) culture samples. This technique is directly relevant for diagnostic laboratories that perform MAP cultures but may also be applicable to the detection of other species, including M. avium and M. tuberculosis. PMID:25609725

  14. New method for ethephon ((2-chloroethyl)phosphonic acid) residue analysis, and detection of residual levels in the fruit and vegetables of Western Japan.

    PubMed

    Takenaka, Shigeyuki

    2002-12-18

    A new method for the detection and quantification of ethephon residues in fruit and vegetables was developed. The present study indicates that fruit and vegetables require a rapid and simple cleanup step before using gas chromatograph/mass spectrometry. The recovery and precision of the new method were evaluated by spiking the fruit and vegetable samples with 0.01-0.1 microg/g of ethephon. The amount of ethephon residue can be determined with good accuracy (recovery, 78.6-109%; coefficient variation, 2.65-6.41%), and the detection limit, defined as the amount of ethephon equivalent to three standard deviations (SD) of the noise level in observations at the baseline level of the selected ion (m/z 110), was 4 pg. The determination limit, defined as the equivalent to 8 SD of the noise level, was 11 pg. The working range was between 10 and 1000 ng/mL, and the correlation coefficient was 0.999 in the five experiments. Ethephon residues were determined between <2 and 97 ng/g in commercial pineapples from Western Japan.

  15. Explosives detection system and method

    DOEpatents

    Reber, Edward L.; Jewell, James K.; Rohde, Kenneth W.; Seabury, Edward H.; Blackwood, Larry G.; Edwards, Andrew J.; Derr, Kurt W.

    2007-12-11

    A method of detecting explosives in a vehicle includes providing a first rack on one side of the vehicle, the rack including a neutron generator and a plurality of gamma ray detectors; providing a second rack on another side of the vehicle, the second rack including a neutron generator and a plurality of gamma ray detectors; providing a control system, remote from the first and second racks, coupled to the neutron generators and gamma ray detectors; using the control system, causing the neutron generators to generate neutrons; and performing gamma ray spectroscopy on spectra read by the gamma ray detectors to look for a signature indicative of presence of an explosive. Various apparatus and other methods are also provided.

  16. Phenol-Sulfuric Acid Method for Total Carbohydrates

    NASA Astrophysics Data System (ADS)

    Nielsen, S. Suzanne

    The phenol-sulfuric acid method is a simple and rapid colorimetric method to determine total carbohydrates in a sample. The method detects virtually all classes of carbohydrates, including mono-, di-, oligo-, and polysaccharides. Although the method detects almost all carbohydrates, the absorptivity of the different carbohydrates varies. Thus, unless a sample is known to contain only one carbohydrate, the results must be expressed arbitrarily in terms of one carbohydrate.

  17. Structure-property study of the Raman spectroscopy detection of fusaric acid and analogs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Food security can benefit from the development of selective methods to detect toxins. Fusaric acid is a mycotoxin produced by certain fungi occasionally found in agricultural commodities. Raman spectroscopy allows selective detection of analytes associated with certain spectral characteristics relat...

  18. Compositions and methods for detecting single nucleotide polymorphisms

    SciTech Connect

    Yeh, Hsin-Chih; Werner, James; Martinez, Jennifer S.

    2016-11-22

    Described herein are nucleic acid based probes and methods for discriminating and detecting single nucleotide variants in nucleic acid molecules (e.g., DNA). The methods include use of a pair of probes can be used to detect and identify polymorphisms, for example single nucleotide polymorphism in DNA. The pair of probes emit a different fluorescent wavelength of light depending on the association and alignment of the probes when hybridized to a target nucleic acid molecule. Each pair of probes is capable of discriminating at least two different nucleic acid molecules that differ by at least a single nucleotide difference. The methods can probes can be used, for example, for detection of DNA polymorphisms that are indicative of a particular disease or condition.

  19. Fluorescent hybridization probes for nucleic acid detection.

    PubMed

    Guo, Jia; Ju, Jingyue; Turro, Nicholas J

    2012-04-01

    Due to their high sensitivity and selectivity, minimum interference with living biological systems, and ease of design and synthesis, fluorescent hybridization probes have been widely used to detect nucleic acids both in vivo and in vitro. Molecular beacons (MBs) and binary probes (BPs) are two very important hybridization probes that are designed based on well-established photophysical principles. These probes have shown particular applicability in a variety of studies, such as mRNA tracking, single nucleotide polymorphism (SNP) detection, polymerase chain reaction (PCR) monitoring, and microorganism identification. Molecular beacons are hairpin oligonucleotide probes that present distinctive fluorescent signatures in the presence and absence of their target. Binary probes consist of two fluorescently labeled oligonucleotide strands that can hybridize to adjacent regions of their target and generate distinctive fluorescence signals. These probes have been extensively studied and modified for different applications by modulating their structures or using various combinations of fluorophores, excimer-forming molecules, and metal complexes. This review describes the applicability and advantages of various hybridization probes that utilize novel and creative design to enhance their target detection sensitivity and specificity.

  20. Development of an ion-pairing reversed-phase liquid chromatography method using a double detection analysis (UV and evaporative light scattering detection) to monitor the stability of Alimta(®)-pemetrexed preparations: identification and quantification of L-glutamic acid as a potential degradation product.

    PubMed

    Respaud, Renaud; Tournamille, Jean-François; Croix, Cécile; Laborie, Hélène; Elfakir, Claire; Viaud-Massuard, Marie-Claude

    2011-01-25

    A new method based on high-performance liquid chromatography coupled to ultraviolet and evaporative light scattering detection (HPLC-UV-ELSD) was developed for the determination of L-glutamic acid, a potential degradation product of pemetrexed, and for the quantification of pemetrexed itself. This is an ion-pairing, reversed-phase method. The column was a Synergi MAX-RP C12 4 μm (150 mm x 4.6 mm). The mobile phase was 1 mM tridecafluoroheptanoic acid in aqueous solution and acetonitrile under gradient elution mode. L-Glutamic acid was detected by ELSD, and pemetrexed by UV at 254 nm. Good resolution was achieved between pemetrexed and L-glutamic acid. The HPLC method was validated according to SFSTP and ICH guidelines, and applied the accuracy profile procedure with a five-level validation experimental design. For pemetrexed, the decision criteria selected consisted of the acceptability limits (±3%) and the proportion of results within the calculated tolerance intervals (95%). In conclusion, the proposed analytical procedures were validated over the selected validation domains for L-glutamic acid (0.005-0.025 mg/mL) and pemetrexed (0.4-0.6 mg/mL) and shown to provide a very effective method.

  1. Method of detecting genetic deletions identified with chromosomal abnormalities

    DOEpatents

    Gray, Joe W; Pinkel, Daniel; Tkachuk, Douglas

    2013-11-26

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyzes. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acids probes are typically of a complexity greater tha 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particlularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML) and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar ut genetically different diseases, and for many prognostic and diagnostic applications.

  2. Method of detecting genetic translocations identified with chromosomal abnormalities

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel; Tkachuk, Douglas

    2001-01-01

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML) and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar but genetically different diseases, and for many prognostic and diagnostic applications.

  3. Well acidizing compositions and method

    SciTech Connect

    Gardener, T.R.; Dill, W.R.; Ford, W.G.F.; King, K.L.

    1991-07-23

    This patent describes a concentrate which forms an acid internal microemulsion well treatment composition when added to an acid treatment fluid. It comprises in the range of from about 20% to about 98% by weight of a hydrocarbon carrier fluid; in the range of from about 1% to about 50% by weight of an alkyl alcohol having in the range of from about 4 to 18 carbon atoms; and in the range of from about 1% to about 50% by weight of an emulsifying agent comprising at least one compound selected from the group consisting of amine salts having ester or amide linkages and propoxylated alcohols, each of the components being different compounds or different mixtures of compounds.

  4. Particle detection systems and methods

    DOEpatents

    Morris, Christopher L.; Makela, Mark F.

    2010-05-11

    Techniques, apparatus and systems for detecting particles such as muons and neutrons. In one implementation, a particle detection system employs a plurality of drift cells, which can be for example sealed gas-filled drift tubes, arranged on sides of a volume to be scanned to track incoming and outgoing charged particles, such as cosmic ray-produced muons. The drift cells can include a neutron sensitive medium to enable concurrent counting of neutrons. The system can selectively detect devices or materials, such as iron, lead, gold, uranium, plutonium, and/or tungsten, occupying the volume from multiple scattering of the charged particles passing through the volume and can concurrently detect any unshielded neutron sources occupying the volume from neutrons emitted therefrom. If necessary, the drift cells can be used to also detect gamma rays. The system can be employed to inspect occupied vehicles at border crossings for nuclear threat objects.

  5. Variation-tolerant capture and multiplex detection of nucleic acids: application to detection of microbes.

    PubMed

    Ohrmalm, Christina; Eriksson, Ronnie; Jobs, Magnus; Simonson, Magnus; Strømme, Maria; Bondeson, Kåre; Herrmann, Björn; Melhus, Asa; Blomberg, Jonas

    2012-10-01

    In contrast to ordinary PCRs, which have a limited multiplex capacity and often return false-negative results due to target variation or inhibition, our new detection strategy, VOCMA (variation-tolerant capture multiplex assay), allows variation-tolerant, target-specific capture and detection of many nucleic acids in one test. Here we demonstrate the use of a single-tube, dual-step amplification strategy that overcomes the usual limitations of PCR multiplexing, allowing at least a 22-plex format with retained sensitivity. Variation tolerance was achieved using long primers and probes designed to withstand variation at known sites and a judicious mix of degeneration and universal bases. We tested VOCMA in situations where enrichment from a large sample volume with high sensitivity and multiplexity is important (sepsis; streptococci, enterococci, and staphylococci, several enterobacteria, candida, and the most important antibiotic resistance genes) and where variation tolerance and high multiplexity is important (gastroenteritis; astrovirus, adenovirus, rotavirus, norovirus genogroups I and II, and sapovirus, as well as enteroviruses, which are not associated with gastroenteritis). Detection sensitivities of 10 to 1,000 copies per reaction were achieved for many targets. VOCMA is a highly multiplex, variation-tolerant, general purpose nucleic acid detection concept. It is a specific and sensitive method for simultaneous detection of nucleic acids from viruses, bacteria, fungi, and protozoa, as well as host nucleic acid, in the same test. It can be run on an ordinary PCR and a Luminex machine and is suitable for both clinical diagnoses and microbial surveillance.

  6. Liquid crystal based biosensors for bile acid detection

    NASA Astrophysics Data System (ADS)

    He, Sihui; Liang, Wenlang; Tanner, Colleen; Fang, Jiyu; Wu, Shin-Tson

    2013-03-01

    The concentration level of bile acids is a useful indicator for early diagnosis of liver diseases. The prevalent measurement method in detecting bile acids is the chromatography coupled with mass spectrometry, which is precise yet expensive. Here we present a biosensor platform based on liquid crystal (LC) films for the detection of cholic acid (CA). This platform has the advantage of low cost, label-free, solution phase detection and simple analysis. In this platform, LC film of 4-Cyano-4'-pentylbiphenyl (5CB) was hosted by a copper grid supported with a polyimide-coated glass substrate. By immersing into sodium dodecyl sulfate (SDS) solution, the LC film was coated with SDS which induced a homeotropic anchoring of 5CB. Addition of CA introduced competitive adsorption between CA and SDS at the interface, triggering a transition from homeotropic to homogeneous anchoring. The detection limit can be tuned by changing the pH value of the solution from 12uM to 170uM.

  7. Method For Detecting Biological Agents

    DOEpatents

    Chen, Liaohai; McBranch, Duncan W.; Wang, Hsing-Lin; Whitten, David G.

    2005-12-27

    A sensor is provided including a polymer capable of having an alterable measurable property from the group of luminescence and electrical conductivity, the polymer having an intermediate combination of a recognition element, a tethering element and a property-altering element bound thereto and capable of altering the measurable property, the intermediate combination adapted for subsequent separation from the polymer upon exposure to an agent having an affinity for binding to the recognition element whereupon the separation of the intermediate combination from the polymer results in a detectable change in the alterable measurable property, and, detecting said detectable change in the alterable measurable property.

  8. Human jagged polypeptide, encoding nucleic acids and methods of use

    DOEpatents

    Li, Linheng; Hood, Leroy

    2000-01-01

    The present invention provides an isolated polypeptide exhibiting substantially the same amino acid sequence as JAGGED, or an active fragment thereof, provided that the polypeptide does not have the amino acid sequence of SEQ ID NO:5 or SEQ ID NO:6. The invention further provides an isolated nucleic acid molecule containing a nucleotide sequence encoding substantially the same amino acid sequence as JAGGED, or an active fragment thereof, provided that the nucleotide sequence does not encode the amino acid sequence of SEQ ID NO:5 or SEQ ID NO:6. Also provided herein is a method of inhibiting differentiation of hematopoietic progenitor cells by contacting the progenitor cells with an isolated JAGGED polypeptide, or active fragment thereof. The invention additionally provides a method of diagnosing Alagille Syndrome in an individual. The method consists of detecting an Alagille Syndrome disease-associated mutation linked to a JAGGED locus.

  9. Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis

    PubMed Central

    Waggoner, Jesse J.; Balassiano, Ilana; Abeynayake, Janaki; Sahoo, Malaya K.; Mohamed-Hadley, Alisha; Liu, Yuanyuan; Vital-Brazil, Juliana Magalhães; Pinsky, Benjamin A.

    2014-01-01

    Background Bacteria of the genus Leptospira, the causative agents of leptospirosis, are categorized into pathogenic and non-pathogenic species. However, the benefit of using a clinical diagnostic that is specific for pathogenic species remains unclear. In this study, we present the development of a real-time PCR (rtPCR) for the detection of pathogenic Leptospira (the pathogenic rtPCR), and we perform a comparison of the pathogenic rtPCR with a published assay that detects all Leptospira species [the undifferentiated febrile illness (UFI) assay] and a reference 16S Leptospira rtPCR, which was originally designed to detect pathogenic species. Methodology/Principal Findings For the pathogenic rtPCR, a new hydrolysis probe was designed for use with primers from the UFI assay, which targets the 16S gene. The pathogenic rtPCR detected Leptospira DNA in 37/37 cultured isolates from 5 pathogenic and one intermediate species. Two strains of the non-pathogenic L. biflexa produced no signal. Clinical samples from 65 patients with suspected leptospirosis were then tested using the pathogenic rtPCR and a reference Leptospira 16S rtPCR. All 65 samples had tested positive for Leptospira using the UFI assay; 62 (95.4%) samples tested positive using the pathogenic rtPCR (p = 0.24). Only 24 (36.9%) samples tested positive in the reference 16S rtPCR (p<0.0001 for comparison with the pathogenic rtPCR and UFI assays). Amplicon sequencing confirmed the detection of pathogenic Leptospira species in 49/50 cases, including 3 cases that were only detected using the UFI assay. Conclusions/Significance The pathogenic rtPCR displayed similar sensitivity to the UFI assay when testing clinical specimens with no difference in specificity. Both assays proved significantly more sensitive than a real-time molecular test used for comparison. Future studies are needed to investigate the clinical and epidemiologic significance of more sensitive Leptospira detection using these tests. PMID:25379890

  10. Detection and isolation of nucleic acid sequences using a bifunctional hybridization probe

    DOEpatents

    Lucas, Joe N.; Straume, Tore; Bogen, Kenneth T.

    2000-01-01

    A method for detecting and isolating a target sequence in a sample of nucleic acids is provided using a bifunctional hybridization probe capable of hybridizing to the target sequence that includes a detectable marker and a first complexing agent capable of forming a binding pair with a second complexing agent. A kit is also provided for detecting a target sequence in a sample of nucleic acids using a bifunctional hybridization probe according to this method.

  11. Nucleic acid sequence detection using multiplexed oligonucleotide PCR

    DOEpatents

    Nolan, John P.; White, P. Scott

    2006-12-26

    Methods for rapidly detecting single or multiple sequence alleles in a sample nucleic acid are described. Provided are all of the oligonucleotide pairs capable of annealing specifically to a target allele and discriminating among possible sequences thereof, and ligating to each other to form an oligonucleotide complex when a particular sequence feature is present (or, alternatively, absent) in the sample nucleic acid. The design of each oligonucleotide pair permits the subsequent high-level PCR amplification of a specific amplicon when the oligonucleotide complex is formed, but not when the oligonucleotide complex is not formed. The presence or absence of the specific amplicon is used to detect the allele. Detection of the specific amplicon may be achieved using a variety of methods well known in the art, including without limitation, oligonucleotide capture onto DNA chips or microarrays, oligonucleotide capture onto beads or microspheres, electrophoresis, and mass spectrometry. Various labels and address-capture tags may be employed in the amplicon detection step of multiplexed assays, as further described herein.

  12. Method for sequencing nucleic acid molecules

    DOEpatents

    Korlach, Jonas; Webb, Watt W.; Levene, Michael; Turner, Stephen; Craighead, Harold G.; Foquet, Mathieu

    2006-05-30

    The present invention is directed to a method of sequencing a target nucleic acid molecule having a plurality of bases. In its principle, the temporal order of base additions during the polymerization reaction is measured on a molecule of nucleic acid, i.e. the activity of a nucleic acid polymerizing enzyme on the template nucleic acid molecule to be sequenced is followed in real time. The sequence is deduced by identifying which base is being incorporated into the growing complementary strand of the target nucleic acid by the catalytic activity of the nucleic acid polymerizing enzyme at each step in the sequence of base additions. A polymerase on the target nucleic acid molecule complex is provided in a position suitable to move along the target nucleic acid molecule and extend the oligonucleotide primer at an active site. A plurality of labelled types of nucleotide analogs are provided proximate to the active site, with each distinguishable type of nucleotide analog being complementary to a different nucleotide in the target nucleic acid sequence. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand at the active site, where the nucleotide analog being added is complementary to the nucleotide of the target nucleic acid at the active site. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The steps of providing labelled nucleotide analogs, polymerizing the growing nucleic acid strand, and identifying the added nucleotide analog are repeated so that the nucleic acid strand is further extended and the sequence of the target nucleic acid is determined.

  13. Method for sequencing nucleic acid molecules

    DOEpatents

    Korlach, Jonas; Webb, Watt W.; Levene, Michael; Turner, Stephen; Craighead, Harold G.; Foquet, Mathieu

    2006-06-06

    The present invention is directed to a method of sequencing a target nucleic acid molecule having a plurality of bases. In its principle, the temporal order of base additions during the polymerization reaction is measured on a molecule of nucleic acid, i.e. the activity of a nucleic acid polymerizing enzyme on the template nucleic acid molecule to be sequenced is followed in real time. The sequence is deduced by identifying which base is being incorporated into the growing complementary strand of the target nucleic acid by the catalytic activity of the nucleic acid polymerizing enzyme at each step in the sequence of base additions. A polymerase on the target nucleic acid molecule complex is provided in a position suitable to move along the target nucleic acid molecule and extend the oligonucleotide primer at an active site. A plurality of labelled types of nucleotide analogs are provided proximate to the active site, with each distinguishable type of nucleotide analog being complementary to a different nucleotide in the target nucleic acid sequence. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand at the active site, where the nucleotide analog being added is complementary to the nucleotide of the target nucleic acid at the active site. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The steps of providing labelled nucleotide analogs, polymerizing the growing nucleic acid strand, and identifying the added nucleotide analog are repeated so that the nucleic acid strand is further extended and the sequence of the target nucleic acid is determined.

  14. A rapid method for preparation of nucleic acid extracts from potato psyllids for detection of 'Candidatus Liberibacter solancearum' and molecular analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A rapid method has been developed and validated for PCR analysis of potato psyllids for Candidatus Liberibacter solanacearum (Lso), the causal agent of zebra chip disease of potatoes. The method is also suitable for PCR amplification and high resolution melting analysis of the cytochrome oxidase I ...

  15. Photodynamic Detection of Peritoneal Metastases Using 5-Aminolevulinic Acid (ALA)

    PubMed Central

    Yonemura, Yutaka; Endo, Yoshio; Canbay, Emel; Liu, Yang; Ishibashi, Haruaki; Mizumoto, Akiyoshi; Hirano, Masamitu; Imazato, Yuuki; Takao, Nobuyuki; Ichinose, Masumi; Noguchi, Kousuke; Li, Yan; Wakama, Satoshi; Yamada, Kazuhiro; Hatano, Koutarou; Shintani, Hiroshi; Yoshitake, Hiroyuki; Ogura, Shun-ichiro

    2017-01-01

    In the past, peritoneal metastasis (PM) was considered as a terminal stage of cancer. From the early 1990s, however, a new comprehensive treatment consisting of cytoreductive surgery and perioperative chemotherapy has been established to improve long-term survival for selected patients with PM. Among prognostic indicators after the treatment, completeness of cytoreduction is the most independent predictors of survival. However, peritoneal recurrence is a main cause of recurrence, even after complete cytoreduction. As a cause of peritoneal recurrence, small PM may be overlooked at the time of cytoreductive surgery (CRS), therefore, development of a new method to detect small PM is desired. Recently, photodynamic diagnosis (PDD) was developed for detection of PM. The objectives of this review were to evaluate whether PDD using 5-aminolevulinic acid (ALA) could improve detection of small PM. PMID:28257041

  16. Photodynamic Detection of Peritoneal Metastases Using 5-Aminolevulinic Acid (ALA).

    PubMed

    Yonemura, Yutaka; Endo, Yoshio; Canbay, Emel; Liu, Yang; Ishibashi, Haruaki; Mizumoto, Akiyoshi; Hirano, Masamitu; Imazato, Yuuki; Takao, Nobuyuki; Ichinose, Masumi; Noguchi, Kousuke; Li, Yan; Wakama, Satoshi; Yamada, Kazuhiro; Hatano, Koutarou; Shintani, Hiroshi; Yoshitake, Hiroyuki; Ogura, Shun-Ichiro

    2017-03-01

    In the past, peritoneal metastasis (PM) was considered as a terminal stage of cancer. From the early 1990s, however, a new comprehensive treatment consisting of cytoreductive surgery and perioperative chemotherapy has been established to improve long-term survival for selected patients with PM. Among prognostic indicators after the treatment, completeness of cytoreduction is the most independent predictors of survival. However, peritoneal recurrence is a main cause of recurrence, even after complete cytoreduction. As a cause of peritoneal recurrence, small PM may be overlooked at the time of cytoreductive surgery (CRS), therefore, development of a new method to detect small PM is desired. Recently, photodynamic diagnosis (PDD) was developed for detection of PM. The objectives of this review were to evaluate whether PDD using 5-aminolevulinic acid (ALA) could improve detection of small PM.

  17. Enhanced Resolution of Chiral Amino Acids with Capillary Electrophoresis for Biosignature Detection in Extraterrestrial Samples.

    PubMed

    Creamer, Jessica S; Mora, Maria F; Willis, Peter A

    2017-01-17

    Amino acids are fundamental building blocks of terrestrial life as well as ubiquitous byproducts of abiotic reactions. In order to distinguish between amino acids formed by abiotic versus biotic processes it is possible to use chemical distributions to identify patterns unique to life. This article describes two capillary electrophoresis methods capable of resolving 17 amino acids found in high abundance in both biotic and abiotic samples (seven enantiomer pairs d/l-Ala, -Asp, -Glu, -His, -Leu, -Ser, -Val and the three achiral amino acids Gly, β-Ala, and GABA). To resolve the 13 neutral amino acids one method utilizes a background electrolyte containing γ-cyclodextrin and sodium taurocholate micelles. The acidic amino acid enantiomers were resolved with γ-cyclodextrin alone. These methods allow detection limits down to 5 nM for the neutral amino acids and 500 nM for acidic amino acids and were used to analyze samples collected from Mono Lake with minimal sample preparation.

  18. Advances in rapid detection methods for foodborne pathogens.

    PubMed

    Zhao, Xihong; Lin, Chii-Wann; Wang, Jun; Oh, Deog Hwan

    2014-03-28

    Food safety is increasingly becoming an important public health issue, as foodborne diseases present a widespread and growing public health problem in both developed and developing countries. The rapid and precise monitoring and detection of foodborne pathogens are some of the most effective ways to control and prevent human foodborne infections. Traditional microbiological detection and identification methods for foodborne pathogens are well known to be time consuming and laborious as they are increasingly being perceived as insufficient to meet the demands of rapid food testing. Recently, various kinds of rapid detection, identification, and monitoring methods have been developed for foodborne pathogens, including nucleic-acid-based methods, immunological methods, and biosensor-based methods, etc. This article reviews the principles, characteristics, and applications of recent rapid detection methods for foodborne pathogens.

  19. Optimization of a peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) method for the detection of bacteria and disclosure of a formamide effect.

    PubMed

    Santos, Rita S; Guimarães, Nuno; Madureira, Pedro; Azevedo, Nuno F

    2014-10-10

    Despite the fact that fluorescence in situ hybridization (FISH) is a well-established technique to identify microorganisms, there is a lack of understanding concerning the interaction of the different factors affecting the obtained fluorescence. In here, we used flow cytometry to study the influence of three essential factors in hybridization - temperature, time and formamide concentration - in an effort to optimize the performance of a Peptide Nucleic Acid (PNA) probe targeting bacteria (EUB338). The PNA-FISH optimization was performed with bacteria representing different families employing response surface methodology. Surprisingly, the optimum concentration of formamide varied according to the bacterium tested. While hybridization on the bacteria possessing the thickest peptidoglycan was more successful at nearly 50% (v/v) formamide, hybridization on all other microorganisms appeared to improve with much lower formamide concentrations. Gram staining and transmission electron microscopy allowed us to confirm that the overall effect of formamide concentration on the fluorescence intensity is a balance between a harmful effect on the bacterial cell envelope, affecting cellular integrity, and the beneficial denaturant effect in the hybridization process. We also conclude that microorganisms belonging to different families will require different hybridization parameters for the same FISH probe, meaning that an optimum universal PNA-FISH procedure is non-existent for these situations.

  20. Enzyme-assisted target recycling (EATR) for nucleic acid detection.

    PubMed

    Gerasimova, Yulia V; Kolpashchikov, Dmitry M

    2014-09-07

    Fast, reliable and sensitive methods for nucleic acid detection are of growing practical interest with respect to molecular diagnostics of cancer, infectious and genetic diseases. Currently, PCR-based and other target amplification strategies are most extensively used in practice. At the same time, such assays have limitations that can be overcome by alternative approaches. There is a recent explosion in the design of methods that amplify the signal produced by a nucleic acid target, without changing its copy number. This review aims at systematization and critical analysis of the enzyme-assisted target recycling (EATR) signal amplification technique. The approach uses nucleases to recognize and cleave the probe-target complex. Cleavage reactions produce a detectable signal. The advantages of such techniques are potentially low sensitivity to contamination and lack of the requirement of a thermal cycler. Nucleases used for EATR include sequence-dependent restriction or nicking endonucleases or sequence independent exonuclease III, lambda exonuclease, RNase H, RNase HII, AP endonuclease, duplex-specific nuclease, DNase I, or T7 exonuclease. EATR-based assays are potentially useful for point-of-care diagnostics, single nucleotide polymorphisms genotyping and microRNA analysis. Specificity, limit of detection and the potential impact of EATR strategies on molecular diagnostics are discussed.

  1. Apparatus for point-of-care detection of nucleic acid in a sample

    DOEpatents

    Bearinger, Jane P.; Dugan, Lawrence C.

    2016-04-19

    Provided herein are methods and apparatus for detecting a target nucleic acid in a sample and related methods and apparatus for diagnosing a condition in an individual. The condition is associated with presence of nucleic acid produced by certain pathogens in the individual.

  2. Method of detecting sulfur dioxide

    DOEpatents

    Spicer, Leonard D.; Bennett, Dennis W.; Davis, Jon F.

    1985-01-01

    (CH.sub.3).sub.3 SiNSO is produced by the reaction of ((CH.sub.3).sub.3 Si).sub.2 NH with SO.sub.2. Also produced in the reaction are ((CH.sub.3).sub.3 Si).sub.2 O and a new solid compound [NH.sub.4 ][(CH.sub.3).sub.3 SiOSO.sub.2 ]. Both (CH.sub.3).sub.3 SiNSO and [NH.sub.4 ][(CH.sub.3).sub.3 SiOSO.sub.2 ] have fluorescent properties. The reaction of the subject invention is used in a method of measuring the concentration of SO.sub.2 pollutants in gases. By the method, a sample of gas is bubbled through a solution of ((CH.sub.3).sub.3 Si).sub.2 NH, whereby any SO.sub.2 present in the gas will react to produce the two fluorescent products. The measured fluorescence of these products can then be used to calculate the concentration of SO.sub.2 in the original gas sample. The solid product [NH.sub.4][(CH.sub.3).sub.3 SiOSO.sub.2 ] may be used as a standard in solid state NMR spectroscopy.

  3. Robust statistical methods for automated outlier detection

    NASA Technical Reports Server (NTRS)

    Jee, J. R.

    1987-01-01

    The computational challenge of automating outlier, or blunder point, detection in radio metric data requires the use of nonstandard statistical methods because the outliers have a deleterious effect on standard least squares methods. The particular nonstandard methods most applicable to the task are the robust statistical techniques that have undergone intense development since the 1960s. These new methods are by design more resistant to the effects of outliers than standard methods. Because the topic may be unfamiliar, a brief introduction to the philosophy and methods of robust statistics is presented. Then the application of these methods to the automated outlier detection problem is detailed for some specific examples encountered in practice.

  4. Methods for analyzing nucleic acid sequences

    DOEpatents

    Korlach, Jonas; Webb, Watt W.; Levene, Michael; Turner, Stephen; Craighead, Harold G.; Foquet, Mathieu

    2011-05-17

    The present invention is directed to a method of sequencing a target nucleic acid. The method provides a complex comprising a polymerase enzyme, a target nucleic acid molecule, and a primer, wherein the complex is immobilized on a support Fluorescent label is attached to a terminal phosphate group of the nucleotide or nucleotide analog. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The time duration of the signal from labeled nucleotides or nucleotide analogs that become incorporated is distinguished from freely diffusing labels by a longer retention in the observation volume for the nucleotides or nucleotide analogs that become incorporated than for the freely diffusing labels.

  5. Phenolic acid esterases, coding sequences and methods

    DOEpatents

    Blum, David L.; Kataeva, Irina; Li, Xin-Liang; Ljungdahl, Lars G.

    2002-01-01

    Described herein are four phenolic acid esterases, three of which correspond to domains of previously unknown function within bacterial xylanases, from XynY and XynZ of Clostridium thermocellum and from a xylanase of Ruminococcus. The fourth specifically exemplified xylanase is a protein encoded within the genome of Orpinomyces PC-2. The amino acids of these polypeptides and nucleotide sequences encoding them are provided. Recombinant host cells, expression vectors and methods for the recombinant production of phenolic acid esterases are also provided.

  6. Method for detecting coliform organisms

    NASA Technical Reports Server (NTRS)

    Nishioka, K.; Nibley, D. A.; Jeffers, E. L.; Brooks, R. L. (Inventor)

    1983-01-01

    A method and apparatus are disclosed for determining the concentration of coliform bacteria in a sample. The sample containing the coliform bacteria is cultured in a liquid growth medium. The cultured bacteria produce hydrogen and the hydrogen is vented to a second cell containing a buffer solution in which the hydrogen dissolves. By measuring the potential change in the buffer solution caused by the hydrogen, as a function of time, the initial concentration of bacteria in the sample is determined. Alternatively, the potential change in the buffer solution can be compared with the potential change in the liquid growth medium to verify that the potential change in the liquid growth medium is produced primarily by the hydrogen gas produced by the coliform bacteria.

  7. Improved moving object detection and tracking method

    NASA Astrophysics Data System (ADS)

    Li, Zhanli; Yang, Fang; Li, Hong-an

    2016-07-01

    At present, the detection and tracking for video moving object have been used in many fields. Aimed at the limitation of the traditional adjacent frame difference method, this paper presented the three-frame difference method to detect objects. For moving object tracking, this paper proposed a method combining Kalman filter and Mean-Shift, and used the prediction function of Kalman filter to overcome the defect of Mean-Shift in selecting the initial position of the candidate object. Experimental results showed that the detection and tracking method proposed in this paper are simple, precise, and perform a good result.

  8. Amino Acid Detection in Cometary Matter?

    NASA Astrophysics Data System (ADS)

    Meierhenrich, U. J.; Munoz Caro, G. M.; Thiemann, W.; Goesmann, F.; Rosenbauer, H.

    2003-04-01

    The recent identification of amino acid structures in interstellar ice analogues [1, 2] strongly supports the assumption that amino acids are abundant in cometary matter too. Cometary matter is assumed to be built up of aggregates of interstellar dust particles. Amino acids are the molecular building blocks of proteins in living organisms. These results amplified the scientific interest in the ESA cometary mission Rosetta. The Rosetta Lander includes the Cosac experiment dedicated to the identification of chiral organic molecules in cometary matter itshape in situ \\upshape by multi column gas chromatography coupled with a reflectron time-of-flight mass spectrometer. However, the envisaged itshape in situ \\upshape amino acid analysis on the cometary surface requires special technical emphasis of the COSAC instrumentation. The context in which the amino acid identification in cometary matter is of interest will be outlined and the analytical solutions that make amino acids accessible to the COSAC instrument will be presented. A succesful identification of amino acid structures in cometary matter would help to understand the beginnings of the biomolecular evolution and the origin of the biomolecular asymmetry. [1] G.M. Muñoz Caro, U.J. Meierhenrich, W.A. Schutte, B. Barbier, A. Arcones Sergovia, H. Rosenbauer, W.H.-P. Thiemann, A. Brack, J.M. Greenberg: itshape Nature \\upshape 416 (2002), 403-406. [2] M.P. Bernstein, J.P. Dworkin, S.A. Sandford, G.W. Cooper, L.J. Allamandola: itshape Nature \\upshape 416 (2002), 401-403.

  9. Spectral analysis method for detecting an element

    DOEpatents

    Blackwood, Larry G [Idaho Falls, ID; Edwards, Andrew J [Idaho Falls, ID; Jewell, James K [Idaho Falls, ID; Reber, Edward L [Idaho Falls, ID; Seabury, Edward H [Idaho Falls, ID

    2008-02-12

    A method for detecting an element is described and which includes the steps of providing a gamma-ray spectrum which has a region of interest which corresponds with a small amount of an element to be detected; providing nonparametric assumptions about a shape of the gamma-ray spectrum in the region of interest, and which would indicate the presence of the element to be detected; and applying a statistical test to the shape of the gamma-ray spectrum based upon the nonparametric assumptions to detect the small amount of the element to be detected.

  10. Rapid qualitative method for detecting staphylococcal nuclease in foods.

    PubMed Central

    Koupal, A; Deibel, R H

    1978-01-01

    A rapid method for the detection of heat-stable staphylococcal nuclease in foods is described. The procedure consists of an acid precipitation, boiling, and centrifugation followed by enzyme detection in an agar plate containing deoxyribonucleic acid. To test the efficacy of the procedure, purified Staphylococcus aureus nuclease was added to various foods and recovery experiments were performed. Additionally, foods were inoculated and incubated with S. aureus, and the staphylococcal counts were compared with nuclease activity. The results indicate that the procedure possesses merit for a rapid method that can be incorporated into quality control programs. The procedure requires approximately 2.5 h, and it will detect nuclease levels as low as 10 ng/g of food. Images PMID:677882

  11. Acid detection by taste receptor cells.

    PubMed

    DeSimone, J A; Lyall, V; Heck, G L; Feldman, G M

    2001-12-01

    Sourness is a primary taste quality that evokes an innate rejection response in humans and many other animals. Acidic stimuli are the unique sources of sour taste so a rejection response may serve to discourage ingestion of foods spoiled by acid producing microorganisms. The investigation of mechanisms by which acids excite taste receptor cells (TRCs) is complicated by wide species variability and within a species, apparently different mechanisms for strong and weak acids. The problem is further complicated by the fact that the receptor cells are polarized epithelial cells with different apical and basolateral membrane properties. The cellular mechanisms proposed for acid sensing in taste cells include, the direct blockage of apical K(+) channels by protons, an H(+)-gated Ca(2+) channel, proton conduction through apical amiloride-blockable Na(+) channels, a Cl(-) conductance blocked by NPPB, the activation of the proton-gated channel, BNC-1, a member of the Na(+) channel/degenerin super family, and by stimulus-evoked changes in intracellular pH. Acid-induced intracellular pH changes appear to be similar to those reported in other mammalian acid-sensing cells, such as type-I cells of the carotid body, and neurons found in the ventrolateral medulla, nucleus of the solitary tract, the medullary raphe, and the locus coceuleus. Like type-I carotid body cells and brainstem neurons, isolated TRCs demonstrate a linear relationship between intracellular pH (pH(i)) and extracellular pH (pH(o)) with slope, DeltapH(i)/DeltapH(o) near unity. Acid-sensing cells also appear to regulate pH(i) when intracellular pH changes occur under iso-extracellular pH conditions, but fail to regulate their pH when pH(i) changes are induced by decreasing extracellular pH. We shall discuss the current status of proposed acid-sensing taste mechanisms, emphasizing pH-tracking in receptor cells.

  12. Detecting sulphate aerosol geoengineering with different methods

    NASA Astrophysics Data System (ADS)

    Lo, Y. T. Eunice; Charlton-Perez, Andrew J.; Lott, Fraser C.; Highwood, Eleanor J.

    2016-12-01

    Sulphate aerosol injection has been widely discussed as a possible way to engineer future climate. Monitoring it would require detecting its effects amidst internal variability and in the presence of other external forcings. We investigate how the use of different detection methods and filtering techniques affects the detectability of sulphate aerosol geoengineering in annual-mean global-mean near-surface air temperature. This is done by assuming a future scenario that injects 5 Tg yr‑1 of sulphur dioxide into the stratosphere and cross-comparing simulations from 5 climate models. 64% of the studied comparisons would require 25 years or more for detection when no filter and the multi-variate method that has been extensively used for attributing climate change are used, while 66% of the same comparisons would require fewer than 10 years for detection using a trend-based filter. This highlights the high sensitivity of sulphate aerosol geoengineering detectability to the choice of filter. With the same trend-based filter but a non-stationary method, 80% of the comparisons would require fewer than 10 years for detection. This does not imply sulphate aerosol geoengineering should be deployed, but suggests that both detection methods could be used for monitoring geoengineering in global, annual mean temperature should it be needed.

  13. Detecting sulphate aerosol geoengineering with different methods.

    PubMed

    Lo, Y T Eunice; Charlton-Perez, Andrew J; Lott, Fraser C; Highwood, Eleanor J

    2016-12-15

    Sulphate aerosol injection has been widely discussed as a possible way to engineer future climate. Monitoring it would require detecting its effects amidst internal variability and in the presence of other external forcings. We investigate how the use of different detection methods and filtering techniques affects the detectability of sulphate aerosol geoengineering in annual-mean global-mean near-surface air temperature. This is done by assuming a future scenario that injects 5 Tg yr(-1) of sulphur dioxide into the stratosphere and cross-comparing simulations from 5 climate models. 64% of the studied comparisons would require 25 years or more for detection when no filter and the multi-variate method that has been extensively used for attributing climate change are used, while 66% of the same comparisons would require fewer than 10 years for detection using a trend-based filter. This highlights the high sensitivity of sulphate aerosol geoengineering detectability to the choice of filter. With the same trend-based filter but a non-stationary method, 80% of the comparisons would require fewer than 10 years for detection. This does not imply sulphate aerosol geoengineering should be deployed, but suggests that both detection methods could be used for monitoring geoengineering in global, annual mean temperature should it be needed.

  14. Detecting sulphate aerosol geoengineering with different methods

    PubMed Central

    Lo, Y. T. Eunice; Charlton-Perez, Andrew J.; Lott, Fraser C.; Highwood, Eleanor J.

    2016-01-01

    Sulphate aerosol injection has been widely discussed as a possible way to engineer future climate. Monitoring it would require detecting its effects amidst internal variability and in the presence of other external forcings. We investigate how the use of different detection methods and filtering techniques affects the detectability of sulphate aerosol geoengineering in annual-mean global-mean near-surface air temperature. This is done by assuming a future scenario that injects 5 Tg yr−1 of sulphur dioxide into the stratosphere and cross-comparing simulations from 5 climate models. 64% of the studied comparisons would require 25 years or more for detection when no filter and the multi-variate method that has been extensively used for attributing climate change are used, while 66% of the same comparisons would require fewer than 10 years for detection using a trend-based filter. This highlights the high sensitivity of sulphate aerosol geoengineering detectability to the choice of filter. With the same trend-based filter but a non-stationary method, 80% of the comparisons would require fewer than 10 years for detection. This does not imply sulphate aerosol geoengineering should be deployed, but suggests that both detection methods could be used for monitoring geoengineering in global, annual mean temperature should it be needed. PMID:27976697

  15. Bioluminescent bioreporter integrated circuit detection methods

    SciTech Connect

    Simpson, Michael L.; Paulus, Michael J.; Sayler, Gary S.; Applegate, Bruce M.; Ripp, Steven A.

    2005-06-14

    Disclosed are monolithic bioelectronic devices comprising a bioreporter and an OASIC. These bioluminescent bioreporter integrated circuit are useful in detecting substances such as pollutants, explosives, and heavy-metals residing in inhospitable areas such as groundwater, industrial process vessels, and battlefields. Also disclosed are methods and apparatus for detection of particular analytes, including ammonia and estrogen compounds.

  16. A GC-ECD method for estimation of free and bound amino acids, gamma-aminobutyric acid, salicylic acid, and acetyl salicylic acid from Solanum lycopersicum (L.).

    PubMed

    Meher, Hari Charan; Gajbhiye, Vijay T; Singh, Ghanendra

    2011-01-01

    A gas chromatograph with electron capture detection method for estimation of selected metabolites--amino acids (free and bound), gamma-aminobutyric acid (GABA), salicylic acid (SA), and acetyl salicylic acid (ASA) from tomato--is reported. The method is based on nitrophenylation of the metabolites by 1-fluoro-2, 4-dinitrobenzene under aqueous alkaline conditions to form dinitophenyl derivatives. The derivatives were stable under the operating conditions of GC. Analysis of bound amino acids comprised perchloric acid precipitation of protein, alkylation (carboxymethylation) with iodoacetic acid, vapor-phase hydrolysis, and derivatization with 1-fluoro-2,4-dinitrobenzene in that order. The metabolites were resolved in 35 min, using a temperature-programmed run. The method is rapid, sensitive, and precise. It easily measured the typical amino acids (aspartate, asparagine, glutamate, glutamine, alanine, leucine, lysine, and phenylalanine) used for identification and quantification of a protein, resolved amino acids of the same mass (leucine and isoleucine), satisfactorily measured sulfur amino acid (methionine, cystine, and cysteine), and quantified GABA, SA, and ASA, as well. The developed method was validated for specificity, linearity, and precision. It has been applied and recommended for estimation of 25 metabolites from Solanum lycopersicum (L.).

  17. CURRENT METHODS FOR DETECTION OF CRYPTOSPORIDIUM SPECIES

    EPA Science Inventory

    Current methods for detecting protozoa in water produce results that are highly variable. It is difficult to determine if the methods themselves, or the procedures for testing these methods, are the source of the variability. If testing procedures are responsible for high varia...

  18. Method and apparatus for detecting halogenated hydrocarbons

    DOEpatents

    Monagle, Matthew; Coogan, John J.

    1997-01-01

    A halogenated hydrocarbon (HHC) detector is formed from a silent discharge (also called a dielectric barrier discharge) plasma generator. A silent discharge plasma device receives a gas sample that may contain one or more HHCs and produces free radicals and excited electrons for oxidizing the HHCs in the gas sample to produce water, carbon dioxide, and an acid including halogens in the HHCs. A detector is used to sensitively detect the presence of the acid. A conductivity cell detector combines the oxidation products with a solvent where dissociation of the acid increases the conductivity of the solvent. The conductivity cell output signal is then functionally related to the presence of HHCs in the gas sample. Other detectors include electrochemical cells, infrared spectrometers, and negative ion mobility spectrometers.

  19. Methodology for detecting residual phosphoric acid in polybenzoxazole fibers.

    PubMed

    Park, Eun Su; Sieber, John; Guttman, Charles; Rice, Kirk; Flynn, Kathleen; Watson, Stephanie; Holmes, Gale

    2009-12-01

    Because of the premature failure of in-service soft-body armor containing the ballistic fiber poly[(benzo-[1,2-d:5,4-d']-benzoxazole-2,6-diyl)-1,4-phenylene] (PBO), the Office of Law Enforcement Standards (OLES) at the National Institute of Standards and Technology (NIST) initiated a research program to investigate the reasons for this failure and to develop testing methodologies and protocols to ensure that these types of failures do not reoccur. In a report that focused on the stability of the benzoxazole ring that is characteristic of PBO fibers, Holmes, G. A.; Rice, K.; Snyder, C. R. J. Mater. Sci. 2006, 41, 4105-4116, showed that the benzoxazole ring was susceptible to hydrolytic degradation under acid conditions. Because of the processing conditions for the fibers, it is suspected by many researchers that residual phosphoric acid may cause degradation of the benzoxazole ring resulting in a reduction of ballistic performance. Prior to this work, no definitive data have indicated the presence of phosphoric acid since the residual phosphorus is not easily extracted and the processed fibers are known to incorporate phosphorus containing processing aids. Methods to efficiently extract phosphorus from PBO are described in this article. Further, characterization determined that the majority of the extractable phosphorus in PBO was attributed to the octyldecyl phosphate processing aid with some phosphoric acid being detected. Analysis by matrix assisted laser desorption ionization of model PBO oligomers indicates that the nonextractable phosphorus is attached to the PBO polymer chain as a monoaryl phosphate ester. The response of model aryl phosphates to NaOH exposure indicates that monoaryl phosphate ester is stable to NaOH washes used in the manufacturing process to neutralize the phosphoric acid reaction medium and to extract residual phosphorus impurities.

  20. Colorimetric Method for Beryllium Surface Contamination Detection

    SciTech Connect

    MCWHORTER, CHRISTOPHER

    2004-03-11

    To address the need for real-time accurate total beryllium analyses, Savannah River Technology Center Analytical Development Section personnel evaluated and modified a colorimetric screening method developed at Los Alamos National Lab to measure beryllium on surfaces. This method was based on a color complex formed by beryllium and chromium azurol s . SRTC converted this visual method to a quantitative analysis method using spectrophotometric detection. The addition of a cationic surfactant (hexadecyltrimethylammonium bromide, CTAB) to the Be-CAS system shifted the complex absorbance away from the CAS absorbance and allowed for the detection. Assuming complete dissolution and a 10 mL rinse solution volume to remove the beryllium from the wipe, the detection limit was calculated comfortably below the free release limit. The spectrophotometric method was rugged and simple enough that it could be used as a field method.

  1. Apparatus and methods for detecting chemical permeation

    DOEpatents

    Vo-Dinh, Tuan

    1994-01-01

    Apparatus and methods for detecting the permeation of hazardous or toxic chemicals through protective clothing are disclosed. The hazardous or toxic chemicals of interest do not possess the spectral characteristic of luminescence. The apparatus and methods utilize a spectrochemical modification technique to detect the luminescence quenching of an indicator compound which upon permeation of the chemical through the protective clothing, the indicator is exposed to the chemical, thus indicating chemical permeation.

  2. Radionuclide detection devices and associated methods

    DOEpatents

    Mann, Nicholas R.; Lister, Tedd E.; Tranter, Troy J.

    2011-03-08

    Radionuclide detection devices comprise a fluid cell comprising a flow channel for a fluid stream. A radionuclide collector is positioned within the flow channel and configured to concentrate one or more radionuclides from the fluid stream onto at least a portion of the radionuclide collector. A scintillator for generating scintillation pulses responsive to an occurrence of a decay event is positioned proximate at least a portion of the radionuclide collector and adjacent to a detection system for detecting the scintillation pulses. Methods of selectively detecting a radionuclide are also provided.

  3. High sensitivity leak detection method and apparatus

    DOEpatents

    Myneni, G.R.

    1994-09-06

    An improved leak detection method is provided that utilizes the cyclic adsorption and desorption of accumulated helium on a non-porous metallic surface. The method provides reliable leak detection at superfluid helium temperatures. The zero drift that is associated with residual gas analyzers in common leak detectors is virtually eliminated by utilizing a time integration technique. The sensitivity of the apparatus of this disclosure is capable of detecting leaks as small as 1 [times] 10[sup [minus]18] atm cc sec[sup [minus]1]. 2 figs.

  4. High sensitivity leak detection method and apparatus

    DOEpatents

    Myneni, Ganapatic R.

    1994-01-01

    An improved leak detection method is provided that utilizes the cyclic adsorption and desorption of accumulated helium on a non-porous metallic surface. The method provides reliable leak detection at superfluid helium temperatures. The zero drift that is associated with residual gas analyzers in common leak detectors is virtually eliminated by utilizing a time integration technique. The sensitivity of the apparatus of this disclosure is capable of detecting leaks as small as 1.times.10.sup.-18 atm cc sec.sup.-1.

  5. Method for remote detection of trace contaminants

    DOEpatents

    Simonson, Robert J.; Hance, Bradley G.

    2003-09-09

    A method for remote detection of trace contaminants in a target area comprises applying sensor particles that preconcentrate the trace contaminant to the target area and detecting the contaminant-sensitive fluorescence from the sensor particles. The sensor particles can have contaminant-sensitive and contaminant-insensitive fluorescent compounds to enable the determination of the amount of trace contaminant present in the target are by relative comparison of the emission of the fluorescent compounds by a local or remote fluorescence detector. The method can be used to remotely detect buried minefields.

  6. Method for predicting peptide detection in mass spectrometry

    DOEpatents

    Kangas, Lars [West Richland, WA; Smith, Richard D [Richland, WA; Petritis, Konstantinos [Richland, WA

    2010-07-13

    A method of predicting whether a peptide present in a biological sample will be detected by analysis with a mass spectrometer. The method uses at least one mass spectrometer to perform repeated analysis of a sample containing peptides from proteins with known amino acids. The method then generates a data set of peptides identified as contained within the sample by the repeated analysis. The method then calculates the probability that a specific peptide in the data set was detected in the repeated analysis. The method then creates a plurality of vectors, where each vector has a plurality of dimensions, and each dimension represents a property of one or more of the amino acids present in each peptide and adjacent peptides in the data set. Using these vectors, the method then generates an algorithm from the plurality of vectors and the calculated probabilities that specific peptides in the data set were detected in the repeated analysis. The algorithm is thus capable of calculating the probability that a hypothetical peptide represented as a vector will be detected by a mass spectrometry based proteomic platform, given that the peptide is present in a sample introduced into a mass spectrometer.

  7. Method for the isolation of citric acid and malic acid in Japanese apricot liqueur for carbon stable isotope analysis.

    PubMed

    Akamatsu, Fumikazu; Hashiguchi, Tomokazu; Hisatsune, Yuri; Oe, Takaaki; Kawao, Takafumi; Fujii, Tsutomu

    2017-02-15

    A method for detecting the undeclared addition of acidic ingredients is required to control the authenticity of Japanese apricot liqueur. We developed an analytical procedure that minimizes carbon isotope discrimination for measurement of the δ(13)C values of citric and malic acid isolated from Japanese apricot liqueur. Our results demonstrated that freeze-drying is preferable to nitrogen spray-drying, because it does not significantly affect the δ(13)C values of citric acid and results in smaller isotope discrimination for malic acid. Both 0.1% formic acid and 0.2% phosphoric acid are acceptable HPLC mobile phases for the isolation of citric and malic acid, although the δ(13)C values of malic acid exhibited relatively large variation compared with citric acid following isolation using either mobile phase. The developed procedure allows precise δ(13)C measurements of citric and malic acid isolated from Japanese apricot liqueur.

  8. Detecting Microbial Nucleic Acids within Nematode Bodies: A Photo Essay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We developed a taxa-specific, fluorescence in situ hybridization (FISH) technique to localize microbial nucleic acids within nematode bodies. This technique involves hybridization of a nucleic acid probe to target microbial sequences. Hybridization is detected microscopically, as the probes have f...

  9. Fluorescence upconversion microbarcodes for multiplexed biological detection: nucleic acid encoding.

    PubMed

    Zhang, Fan; Shi, Qihui; Zhang, Yichi; Shi, Yifeng; Ding, Kunlun; Zhao, Dongyuan; Stucky, Galen D

    2011-09-01

    Fluoride rare-earth-doped upconversion microbarcodes have been successfully developed for multiplexed signaling and nucleic-acid encoding. This kind of novel barcode material can be used for rapid and sensitive analysis of nucleic acids and antigens, which would have many potential applications in clinical, food, and environment detection.

  10. Method and apparatus for staining immobilized nucleic acids

    DOEpatents

    Ramsey, J. Michael; Foote, Robert S.; Jacobson, Stephen C.

    2000-01-01

    A method for staining immobilized nucleic acids includes the steps of affixing DNA probes to a solid substrate, moving target DNA material into proximity with the DNA probes, whereby the target DNA hybridized with specific ones of the DNA probes, and moving a fluorescent dye into proximity with the hybridized target DNA, whereby the fluorescent dye binds to the hybridized DNA to enable subsequent detection of fluorescence.

  11. [Detection method for blurred regions in radiographs].

    PubMed

    Muroi, Tomoya; Lee, Yongbum; Tsai, Du-Yih; Tsurumaki, Masaki

    2014-03-01

    In this paper, we propose a detection method for blurred regions in radiographs. The method involves edge detection using a Sobel filter, manually determining the region of interest (ROI), feature calculation, and classification using a support vector machine. We applied our method to 14 phantom images (7 normal images, 7 blurred images) and 14 clinical images (12 normal images, 2 blurred images). As a result, the average classification accuracies of ROIs with blurring and ROIs without blurring were 98% and 90% for phantom images and clinical images, respectively.

  12. An enhanced Monte Carlo outlier detection method.

    PubMed

    Zhang, Liangxiao; Li, Peiwu; Mao, Jin; Ma, Fei; Ding, Xiaoxia; Zhang, Qi

    2015-09-30

    Outlier detection is crucial in building a highly predictive model. In this study, we proposed an enhanced Monte Carlo outlier detection method by establishing cross-prediction models based on determinate normal samples and analyzing the distribution of prediction errors individually for dubious samples. One simulated and three real datasets were used to illustrate and validate the performance of our method, and the results indicated that this method outperformed Monte Carlo outlier detection in outlier diagnosis. After these outliers were removed, the value of validation by Kovats retention indices and the root mean square error of prediction decreased from 3.195 to 1.655, and the average cross-validation prediction error decreased from 2.0341 to 1.2780. This method helps establish a good model by eliminating outliers. © 2015 Wiley Periodicals, Inc.

  13. A method to attenuate U(VI) mobility in acidic waste plumes using humic acids

    SciTech Connect

    Wan, J.; Dong, W.; Tokunaga, T.K.

    2011-02-01

    Acidic uranium (U) contaminated plumes have resulted from acid-extraction of plutonium during the Cold War and from U mining and milling operations. A sustainable method for in-situ immobilization of U under acidic conditions is not yet available. Here, we propose to use humic acids (HAs) for in-situ U immobilization in acidic waste plumes. Our laboratory batch experiments show that HA can adsorb onto aquifer sediments rapidly, strongly and practically irreversibly. Adding HA greatly enhanced U adsorption capacity to sediments at pH below 5.0. Our column experiments using historically contaminated sediments from the Savannah River Site under slow flow rates (120 and 12 m/y) show that desorption of U and HA were non-detectable over 100 pore-volumes of leaching with simulated acidic groundwaters. Upon HA-treatment, 99% of the contaminant [U] was immobilized at pH < 4.5, compared to 5% and 58% immobilized in the control columns at pH 3.5 and 4.5, respectively. These results demonstrated that HA-treatment is a promising in-situ remediation method for acidic U waste plumes. As a remediation reagent, HAs are resistant to biodegradation, cost effective, nontoxic, and easily introducible to the subsurface.

  14. Nucleic Acid-based Detection of Bacterial Pathogens Using Integrated Microfluidic Platform Systems

    PubMed Central

    Lui, Clarissa; Cady, Nathaniel C.; Batt, Carl A.

    2009-01-01

    The advent of nucleic acid-based pathogen detection methods offers increased sensitivity and specificity over traditional microbiological techniques, driving the development of portable, integrated biosensors. The miniaturization and automation of integrated detection systems presents a significant advantage for rapid, portable field-based testing. In this review, we highlight current developments and directions in nucleic acid-based micro total analysis systems for the detection of bacterial pathogens. Recent progress in the miniaturization of microfluidic processing steps for cell capture, DNA extraction and purification, polymerase chain reaction, and product detection are detailed. Discussions include strategies and challenges for implementation of an integrated portable platform. PMID:22412335

  15. Acetic acid chromoendoscopy: Improving neoplasia detection in Barrett's esophagus

    PubMed Central

    Chedgy, Fergus J Q; Subramaniam, Sharmila; Kandiah, Kesavan; Thayalasekaran, Sreedhari; Bhandari, Pradeep

    2016-01-01

    Barrett’s esophagus (BE) is an important condition given its significant premalignant potential and dismal five-year survival outcomes of advanced esophageal adenocarcinoma. It is therefore suggested that patients with a diagnosis of BE undergo regular surveillance in order to pick up dysplasia at an earlier stage to improve survival. Current “gold-standard” surveillance protocols suggest targeted biopsy of visible lesions followed by four quadrant random biopsies every 2 cm. However, this method of Barrett’s surveillance is fraught with poor endoscopist compliance as the procedures are time consuming and poorly tolerated by patients. There are also significant miss-rates with this technique for the detection of neoplasia as only 13% of early neoplastic lesions appear as visible nodules. Despite improvements in endoscope resolution these problems persist. Chromoendoscopy is an extremely useful adjunct to enhance mucosal visualization and characterization of Barrett’s mucosa. Acetic acid chromoendoscopy (AAC) is a simple, non-proprietary technique that can significantly improve neoplasia detection rates. This topic highlight summarizes the current evidence base behind AAC for the detection of neoplasia in BE and provides an insight into the direction of travel for further research in this area. PMID:27433088

  16. Novel Methods for Detecting Buried Explosive Devices

    DTIC Science & Technology

    2007-04-10

    NQR ), and semiotic data fusion. Bioreporter bacteria look promising for third-world humanitarian applications; they are inexpensive, and...demining, NQR is a promising method for detecting explosive substances; of 50,000 substances that have been tested, none has an NQR signature that can be...approach to a cheap mine detector for humanitarian use. Real-time wavelet processing appears to be a key to extending NQR bomb detection into mine

  17. Method of Fault Detection and Rerouting

    NASA Technical Reports Server (NTRS)

    Medelius, Pedro J. (Inventor); Gibson, Tracy L. (Inventor); Lewis, Mark E. (Inventor)

    2013-01-01

    A system and method for detecting damage in an electrical wire, including delivering at least one test electrical signal to an outer electrically conductive material in a continuous or non-continuous layer covering an electrically insulative material layer that covers an electrically conductive wire core. Detecting the test electrical signals in the outer conductive material layer to obtain data that is processed to identify damage in the outer electrically conductive material layer.

  18. Automated macromolecular crystal detection system and method

    DOEpatents

    Christian, Allen T.; Segelke, Brent; Rupp, Bernard; Toppani, Dominique

    2007-06-05

    An automated macromolecular method and system for detecting crystals in two-dimensional images, such as light microscopy images obtained from an array of crystallization screens. Edges are detected from the images by identifying local maxima of a phase congruency-based function associated with each image. The detected edges are segmented into discrete line segments, which are subsequently geometrically evaluated with respect to each other to identify any crystal-like qualities such as, for example, parallel lines, facing each other, similarity in length, and relative proximity. And from the evaluation a determination is made as to whether crystals are present in each image.

  19. Detection methods for autologous blood doping.

    PubMed

    Segura, J; Monfort, N; Ventura, R

    2012-11-01

    The use of blood doping is forbidden by the World Anti-Doping Agency. Several practices, such as blood transfusions are used to increase oxygen delivery to muscles and all of them are highly pursued. In this regard, the development of accurate methodologies for detecting these prohibited practices is one of the current aims of the anti-doping control laboratories. Flow cytometry methods are able to detect allogeneic blood transfusions but there is no official methodology available to detect autologous blood transfusions. This paper reviews protocols, including the Athlete Biological Passport, that use indirect markers to detect misuse of blood transfusions, especially autologous blood transfusions. The methods of total haemoglobin mass measurements and the detection of metabolites of blood bags plasticizers in urine are reviewed. The latter seems to be an important step forward because it is a fast screening method and it is based on urine, a fluid widely available for doping control. Other innovative approaches to blood transfusion detection are also mentioned. A combination of the reported methodologies and the implementation of the Athlete Biological Passport is becoming a promising approach.

  20. Method for analyzing nucleic acids by means of a substrate having a microchannel structure containing immobilized nucleic acid probes

    DOEpatents

    Ramsey, J. Michael; Foote, Robert S.

    2002-01-01

    A method and apparatus for analyzing nucleic acids includes immobilizing nucleic probes at specific sites within a microchannel structure and moving target nucleic acids into proximity to the probes in order to allow hybridization and fluorescence detection of specific target sequences.

  1. Method for analyzing nucleic acids by means of a substrate having a microchannel structure containing immobilized nucleic acid probes

    DOEpatents

    Ramsey, J. Michael; Foote, Robert S.

    2003-12-09

    A method and apparatus for analyzing nucleic acids includes immobilizing nucleic probes at specific sites within a microchannel structure and moving target nucleic acids into proximity to the probes in order to allow hybridization and fluorescence detection of specific target sequences.

  2. Detection of folic acid protein in human serum using reduced graphene oxide electrodes modified by folic-acid.

    PubMed

    He, Lijie; Wang, Qian; Mandler, Daniel; Li, Musen; Boukherroub, Rabah; Szunerits, Sabine

    2016-01-15

    The detection of disease markers is considered an important step for early diagnosis of cancer. We design in this work a novel electrochemical sensing platform for the sensitive and selective detection of folic acid protein (FP). The platform is fabricated by electrophoretic deposition (EPD) of reduced graphene oxide (rGO) onto a gold electrode and post-functionalization of rGO with folic acid. Upon FP binding, a significant current decrease can be measured using differential pulse voltammetry (DPV). Using this scheme, a detection limit of 1pM is achieved. Importantly, the method also allows the detection of FP in serum being thus an appealing approach for the sensitive detection of biomarkers in clinical samples.

  3. Three Methods of Detection of Hydrazines

    NASA Technical Reports Server (NTRS)

    Griffin, Timothy; Berger, Cristina

    2010-01-01

    Three proposed methods for measuring trace quantities of hydrazines involve ionization and detection of hydrazine derivatives. These methods are intended to overcome the limitations of prior hydrazine- detection methods. Hydrazine (Hz), monomethylhydrazine (MMH), and unsymmetrical dimethylhydrazine (UDMH) are hypergolic fuels and are highly reactive, toxic, and corrosive. A capability to measure concentrations of hydrazines is desirable for detecting leaks and ensuring safety in aerospace settings and in some industrial settings in which these compounds are used. One of the properties (high reactivity) that make it desirable to detect trace amounts of hydrazines also makes it difficult to detect hydrazines and measure their concentrations accurately using prior methods: significant amounts are lost to thermal and catalytic decomposition prior to detection. Further complications arise from the sticky nature of hydrazines: Sample hydrazine molecules tend to become irreversibly adsorbed onto solid surfaces with which they come into contact during transport to detectors, giving rise to drift in detector responses. In each proposed method, the reactive, sticky nature of hydrazines would be turned to advantage by providing a suitably doped substrate surface with which the hydrazines would react. The resulting hydrazine derivatives would be sufficiently less sticky and sufficiently more stable so that fewer molecules would be lost to decomposition or adsorption during transport. Consequently, it would be possible to measure concentration with more sensitivity and less error than in prior techniques. The first proposed method calls for the use of a recently developed technique known as desorption electrospray ionization (DESI), in which a pneumatically assisted micro -electrospray at ambient pressure is directed at a surface of interest. In this case, the surface of interest would be that of a substrate described above.

  4. Apparatus and methods for detecting chemical permeation

    DOEpatents

    Vo-Dinh, T.

    1994-12-27

    Apparatus and methods for detecting the permeation of hazardous or toxic chemicals through protective clothing are disclosed. The hazardous or toxic chemicals of interest do not possess the spectral characteristic of luminescence. The apparatus and methods utilize a spectrochemical modification technique to detect the luminescence quenching of an indicator compound which upon permeation of the chemical through the protective clothing, the indicator is exposed to the chemical, thus indicating chemical permeation. The invention also relates to the fabrication of protective clothing materials. 13 figures.

  5. Method for the production of dicarboxylic acids

    DOEpatents

    Nghiem, Nhuan Phu; Donnelly, Mark; Millard, Cynthia S.; Stols, Lucy

    1999-01-01

    The present invention is an economical fermentation method for the production of carboxylic acids comprising the steps of a) inoculating a medium having a carbon source with a carboxylic acid-producing organism; b) incubating the carboxylic acid-producing organism in an aerobic atmosphere to promote rapid growth of the organism thereby increasing the biomass of the organism; c) controllably releasing oxygen to maintain the aerobic atmosphere; d) controllably feeding the organism having increased biomass with a solution containing the carbon source to maintain the concentration of the carbon source within the medium of about 0.5 g/L up to about 1 g/L; e) depriving the aerobic atmosphere of oxygen to produce an anaerobic atmosphere to cause the organism to undergo anaerobic metabolism; f) controllably feeding the organism having increased biomass a solution containing the carbon source to maintain the concentration of the carbon source within the medium of .gtoreq.1 g/L; and g) converting the carbon source to carboxylic acids using the anaerobic metabolism of the organism.

  6. Method for the production of dicarboxylic acids

    DOEpatents

    Nghiem, N.P.; Donnelly, M.; Millard, C.S.; Stols, L.

    1999-02-09

    The present invention is an economical fermentation method for the production of carboxylic acids comprising the steps of (a) inoculating a medium having a carbon source with a carboxylic acid-producing organism; (b) incubating the carboxylic acid-producing organism in an aerobic atmosphere to promote rapid growth of the organism thereby increasing the biomass of the organism; (c) controllably releasing oxygen to maintain the aerobic atmosphere; (d) controllably feeding the organism having increased biomass with a solution containing the carbon source to maintain the concentration of the carbon source within the medium of about 0.5 g/l up to about 1 g/l; (e) depriving the aerobic atmosphere of oxygen to produce an anaerobic atmosphere to cause the organism to undergo anaerobic metabolism; (f) controllably feeding the organism having increased biomass a solution containing the carbon source to maintain the concentration of the carbon source within the medium of {>=}1 g/l; and (g) converting the carbon source to carboxylic acids using the anaerobic metabolism of the organism. 7 figs.

  7. Determination of selected fatty acids in dried sweat spot using gas chromatography with flame ionization detection.

    PubMed

    Kanďár, Roman; Drábková, Petra; Andrlová, Lenka; Kostelník, Adam; Čegan, Alexander

    2016-11-01

    A method is described for the determination of fatty acids in dried sweat spot and plasma samples using gas chromatography with flame ionization detection. Plasma and dried sweat spot samples were obtained from a group of blood donors. The sweat was collected from each volunteer during exercise. Sweat was spotted onto collection paper containing butylated hydroxytoluene. Fatty acids were derivatized with acetyl chloride in methanol to form methyl esters of fatty acids. The fatty acids in dried sweat spot samples treated with butylated hydroxytoluene and stored at -20°C were stable for 3 months. Our results indicate that sweat contains, among fatty acids with short chain, also fatty acids with long chain and unsaturated fatty acids. Linear relationships between percentage content of selected fatty acids in dried sweat spot and plasma were observed.

  8. An Optical Test Strip for the Detection of Benzoic Acid in Food

    PubMed Central

    Hamzah, Hairul Hisham; Yusof, Nor Azah; Salleh, Abu Bakar; Bakar, Fatimah Abu

    2011-01-01

    Fabrication of a test strip for detection of benzoic acid was successfully implemented by immobilizing tyrosinase, phenol and 3-methyl-2-benzothiazolinone hydrazone (MBTH) onto filter paper using polystyrene as polymeric support. The sensing scheme was based on the decreasing intensity of the maroon colour of the test strip when introduced into benzoic acid solution. The test strip was characterized using optical fiber reflectance and has maximum reflectance at 375 nm. It has shown a highly reproducible measurement of benzoic acid with a calculated RSD of 0.47% (n = 10). The detection was optimized at pH 7. A linear response of the biosensor was obtained in 100 to 700 ppm of benzoic acid with a detection limit (LOD) of 73.6 ppm. At 1:1 ratio of benzoic acid to interfering substances, the main interfering substance is boric acid. The kinetic analyses show that, the inhibition of benzoic is competitive inhibitor and the inhibition constant (Ki) is 52.9 ppm. The activity of immobilized tyrosinase, phenol, and MBTH in the test strip was fairly sustained during 20 days when stored at 3 °C. The developed test strip was used for detection of benzoic acid in food samples and was observed to have comparable results to the HPLC method, hence the developed test strip can be used as an alternative to HPLC in detecting benzoic acid in food products. PMID:22164018

  9. Novel methods for detecting buried explosive devices

    SciTech Connect

    Kercel, S.W.; Burlage, R.S.; Patek, D.R.; Smith, C.M.; Hibbs, A.D.; Rayner, T.J.

    1997-04-01

    Oak Ridge National Laboratory (ORNL) and Quantum Magnetics, Inc. (QM) are exploring novel landmine detection technologies. Technologies considered here include bioreporter bacteria, swept acoustic resonance, nuclear quadrupole resonance (NQR), and semiotic data fusion. Bioreporter bacteria look promising for third-world humanitarian applications; they are inexpensive, and deployment does not require high-tech methods. Swept acoustic resonance may be a useful adjunct to magnetometers in humanitarian demining. For military demining, NQR is a promising method for detecting explosive substances; of 50,000 substances that have been tested, none has an NQR signature that can be mistaken for RDX or TNT. For both military and commercial demining, sensor fusion entails two daunting tasks, identifying fusible features in both present-day and emerging technologies, and devising a fusion algorithm that runs in real-time on cheap hardware. Preliminary research in these areas is encouraging. A bioreporter bacterium for TNT detection is under development. Investigation has just started in swept acoustic resonance as an approach to a cheap mine detector for humanitarian use. Real-time wavelet processing appears to be a key to extending NQR bomb detection into mine detection, including TNT-based mines. Recent discoveries in semiotics may be the breakthrough that will lead to a robust fused detection scheme.

  10. Methods for detection of irradiation of spices.

    PubMed

    Sjöberg, A M; Manninen, M; Härmälä, P; Pinnioja, S

    1990-02-01

    Three types of methods for the identification of irradiation of spices were tested as potential control methods. The methods were microbiological, combining a direct epifluorescent filter technique (DEFT) with a total aerobic plate count (APC), a chemiluminescence method and chemical gas-chromatographic (GC) and GC mass-spectrometric (MS) methods for analysis of volatile oils of spices isolated by steam distillation. Twelve samples of spices, mainly peppers, were analysed before and after gamma-irradiation with doses of 10 and 50 kGy. The chemiluminescence measurements were performed before the irradiation and 10 and 100 days after the irradiation. The best methods for control purposes were the microbiological (DEFT + APC) methods combined with chemiluminescence measurements. No differences were detected between the irradiated and non-irradiated samples with the chemical methods.

  11. Antibodies specific for nucleic acids and applications in genomic detection and clinical diagnostics.

    PubMed

    Hu, Zonglin; Leppla, Stephen H; Li, Baoguang; Elkins, Christopher A

    2014-09-01

    Detection of nucleic acids using antibodies is uncommon. This is in part because nucleic acids are poor immunogens and it is difficult to elicit antibodies having high affinity to each type of nucleic acid while lacking cross-reactivity to others. We describe the origins and applications of a variety of anti-nucleic acid antibodies, including ones reacting with modified nucleosides and nucleotides, single-stranded DNA, double-stranded DNA, RNA, DNA:RNA hybrids, locked-nucleic acids or peptide nucleic acid:nucleic acid hybrids. Carefully selected antibodies can be excellent reagents for detecting bacteria, viruses, small RNAs, microRNAs, R-loops, cancer cells, stem cells, apoptotic cells and so on. The detection may be sensitive, simple, rapid, specific, reproducible, quantitative and cost-effective. Current microarray and diagnostic methods that depend on cDNA or cRNA can be replaced by using antibody detection of nucleic acids. Therefore, development should be encouraged to explore new utilities and create a robust arsenal of new anti-nucleic acid antibodies.

  12. Detection and isolation of nucleic acid sequences using competitive hybridization probes

    DOEpatents

    Lucas, J.N.; Straume, T.; Bogen, K.T.

    1997-04-01

    A method for detecting a target nucleic acid sequence in a sample is provided using hybridization probes which competitively hybridize to a target nucleic acid. According to the method, a target nucleic acid sequence is hybridized to first and second hybridization probes which are complementary to overlapping portions of the target nucleic acid sequence, the first hybridization probe including a first complexing agent capable of forming a binding pair with a second complexing agent and the second hybridization probe including a detectable marker. The first complexing agent attached to the first hybridization probe is contacted with a second complexing agent, the second complexing agent being attached to a solid support such that when the first and second complexing agents are attached, target nucleic acid sequences hybridized to the first hybridization probe become immobilized on to the solid support. The immobilized target nucleic acids are then separated and detected by detecting the detectable marker attached to the second hybridization probe. A kit for performing the method is also provided. 7 figs.

  13. Detection and isolation of nucleic acid sequences using competitive hybridization probes

    DOEpatents

    Lucas, Joe N.; Straume, Tore; Bogen, Kenneth T.

    1997-01-01

    A method for detecting a target nucleic acid sequence in a sample is provided using hybridization probes which competitively hybridize to a target nucleic acid. According to the method, a target nucleic acid sequence is hybridized to first and second hybridization probes which are complementary to overlapping portions of the target nucleic acid sequence, the first hybridization probe including a first complexing agent capable of forming a binding pair with a second complexing agent and the second hybridization probe including a detectable marker. The first complexing agent attached to the first hybridization probe is contacted with a second complexing agent, the second complexing agent being attached to a solid support such that when the first and second complexing agents are attached, target nucleic acid sequences hybridized to the first hybridization probe become immobilized on to the solid support. The immobilized target nucleic acids are then separated and detected by detecting the detectable marker attached to the second hybridization probe. A kit for performing the method is also provided.

  14. Flow cytometric detection method for DNA samples

    DOEpatents

    Nasarabadi,Shanavaz; Langlois, Richard G.; Venkateswaran, Kodumudi S.

    2011-07-05

    Disclosed herein are two methods for rapid multiplex analysis to determine the presence and identity of target DNA sequences within a DNA sample. Both methods use reporting DNA sequences, e.g., modified conventional Taqman.RTM. probes, to combine multiplex PCR amplification with microsphere-based hybridization using flow cytometry means of detection. Real-time PCR detection can also be incorporated. The first method uses a cyanine dye, such as, Cy3.TM., as the reporter linked to the 5' end of a reporting DNA sequence. The second method positions a reporter dye, e.g., FAM.TM. on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA.TM., on the 5' end.

  15. Flow cytometric detection method for DNA samples

    DOEpatents

    Nasarabadi, Shanavaz; Langlois, Richard G.; Venkateswaran, Kodumudi S.

    2006-08-01

    Disclosed herein are two methods for rapid multiplex analysis to determine the presence and identity of target DNA sequences within a DNA sample. Both methods use reporting DNA sequences, e.g., modified conventional Taqman.RTM. probes, to combine multiplex PCR amplification with microsphere-based hybridization using flow cytometry means of detection. Real-time PCR detection can also be incorporated. The first method uses a cyanine dye, such as, Cy3.TM., as the reporter linked to the 5' end of a reporting DNA sequence. The second method positions a reporter dye, e.g., FAM, on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA, on the 5' end.

  16. Objective Method for Pain Detection/Diagnosis

    DTIC Science & Technology

    2013-11-01

    Diagnosis PRINCIPAL INVESTIGATOR: Bor-rong Chen, PhD CONTRACTING ORGANIZATION...REPORT TYPE Final 3. DATES COVERED 15 May 2013 - 15 November 2013 4. TITLE AND SUBTITLE Objective Method for Pain Detection/ Diagnosis 5a...Design of MoPASS System Architecture MoPASS Sensor Platform Development Streaming Firmware MoPASS PC Software MoPASS Desktop Application

  17. Method and apparatus for detecting an analyte

    DOEpatents

    Allendorf, Mark D [Pleasanton, CA; Hesketh, Peter J [Atlanta, GA

    2011-11-29

    We describe the use of coordination polymers (CP) as coatings on microcantilevers for the detection of chemical analytes. CP exhibit changes in unit cell parameters upon adsorption of analytes, which will induce a stress in a static microcantilever upon which a CP layer is deposited. We also describe fabrication methods for depositing CP layers on surfaces.

  18. Method for detecting gas turbine engine flashback

    DOEpatents

    Singh, Kapil Kumar; Varatharajan, Balachandar; Kraemer, Gilbert Otto; Yilmaz, Ertan; Lacy, Benjamin Paul

    2012-09-04

    A method for monitoring and controlling a gas turbine, comprises predicting frequencies of combustion dynamics in a combustor using operating conditions of a gas turbine, receiving a signal from a sensor that is indicative of combustion dynamics in the combustor, and detecting a flashback if a frequency of the received signal does not correspond to the predicted frequencies.

  19. Morphological detection method with girdle structure

    NASA Astrophysics Data System (ADS)

    Chen, Yong; Yan, Gao-shi; Huo, Yu

    2013-09-01

    A flat structure element is usually used in the traditional morphological detection. In this processing method, the infrared target is regarded as a single point without considering the imaging characteristics of infrared dim and small target. There is a gray transition zone between target and background. It is unreasonable that the transition region processed as background in the traditional morphological method. So, the infrared dim target detection results are not ideal by using the traditional morphological processing method. Aimed at this problem, the imaging characteristic of infrared dim and small target is analyzed. The Spatial distribution of infrared target gray scale is calculated. The result shows it is a tip package structure. The top peak is the target. Based on theoretical research on the morphological detection, the girdle structure elements are designed. This structure is composed of two circles. The target neighborhood zones are protected in this structure. They do not participate in the morphological calculation. The sixteen external zones are only participated in the calculation. The morphology of infrared target detection method is established based on this neighborhood zoned structure. The designed girdle structure is used in the opening operation and the traditional flat structure is used in the closing operation. The traditional Top-Hat algorithm is improved according to the girdle structure morphology method. And used the real infrared target image, the improved algorithm is simulated. The processed result shows that the girdle structure morphology method is effective in the background noise restraining. In order to evaluate the image processed result quantitatively, the signal noise ratio and signal noise ratio gain factor are used. Accordingly to the calculated result, the improved algorithm compared with the traditional morphological methods, inhibition of complex background with better.

  20. Morphological boundary detecting method in cineangiogram image

    NASA Astrophysics Data System (ADS)

    Gui, Feng; Lin, QiWei

    2002-04-01

    Atherosclerotic coronary heart disease (CHD) is one of the commonest diseases that is heavily hazardous to people's health. Wall motion abnormalities of L.V. due to myocardia ischeamia caused by coronary atherosclerosis is a significant feature of CHD. This paper was designed to build up a foundation for automatic detection of L.V. contours according to the features of L.V. cineangiograms, for a further study of L.V. wall motion abnormalities. An algorithm that based on morphology for L.V. contours extracting was developed in this paper. As we know morphology is a kind of technique based upon set theory and it can be used for binary image and gray image processing. The principle and the geometrical meaning of morphological boundary detecting for image were discussed in this paper, and the selection of structuring element was analyzed. Comparison was made between morphological boundary detecting and traditional boundary detecting method, conclusion that morphological boundary detecting method has better compatibility and anti-interference capability was reached.

  1. Fault detection with principal component pursuit method

    NASA Astrophysics Data System (ADS)

    Pan, Yijun; Yang, Chunjie; Sun, Youxian; An, Ruqiao; Wang, Lin

    2015-11-01

    Data-driven approaches are widely applied for fault detection in industrial process. Recently, a new method for fault detection called principal component pursuit(PCP) is introduced. PCP is not only robust to outliers, but also can accomplish the objectives of model building, fault detection, fault isolation and process reconstruction simultaneously. PCP divides the data matrix into two parts: a fault-free low rank matrix and a sparse matrix with sensor noise and process fault. The statistics presented in this paper fully utilize the information in data matrix. Since the low rank matrix in PCP is similar to principal components matrix in PCA, a T2 statistic is proposed for fault detection in low rank matrix. And this statistic can illustrate that PCP is more sensitive to small variations in variables than PCA. In addition, in sparse matrix, a new monitored statistic performing the online fault detection with PCP-based method is introduced. This statistic uses the mean and the correlation coefficient of variables. Monte Carlo simulation and Tennessee Eastman (TE) benchmark process are provided to illustrate the effectiveness of monitored statistics.

  2. Method and system for detecting an explosive

    DOEpatents

    Reber, Edward L.; Rohde, Kenneth W.; Blackwood, Larry G.

    2010-12-07

    A method and system for detecting at least one explosive in a vehicle using a neutron generator and a plurality of NaI detectors. Spectra read from the detectors is calibrated by performing Gaussian peak fitting to define peak regions, locating a Na peak and an annihilation peak doublet, assigning a predetermined energy level to one peak in the doublet, and predicting a hydrogen peak location based on a location of at least one peak of the doublet. The spectra are gain shifted to a common calibration, summed for respective groups of NaI detectors, and nitrogen detection analysis performed on the summed spectra for each group.

  3. Detection methods for centrifugal microfluidic platforms.

    PubMed

    Burger, Robert; Amato, Letizia; Boisen, Anja

    2016-02-15

    Centrifugal microfluidics has attracted much interest from academia as well as industry, since it potentially offers solutions for affordable, user-friendly and portable biosensing. A wide range of so-called fluidic unit operations, e.g. mixing, metering, liquid routing, and particle separation, have been developed and allow automation and integration of complex assay protocols in lab-on-a-disc systems. Besides liquid handling, the detection strategy for reading out the assay is crucial for developing a fully integrated system. In this review, we focus on biosensors and readout methods for the centrifugal microfluidics platform and cover optical as well as mechanical and electrical detection principles.

  4. Isothermal amplification detection of nucleic acids by a double-nicked beacon.

    PubMed

    Shi, Chao; Zhou, Meiling; Pan, Mei; Zhong, Guilin; Ma, Cuiping

    2016-03-01

    Isothermal and rapid amplification detection of nucleic acids is an important technology in environmental monitoring, foodborne pathogen detection, and point-of-care clinical diagnostics. Here we have developed a novel method of isothermal signal amplification for single-stranded DNA (ssDNA) detection. The ssDNA target could be used as an initiator, coupled with a double-nicked molecular beacon, to originate amplification cycles, achieving cascade signal amplification. In addition, the method showed good specificity and strong anti-jamming capability. Overall, it is a one-pot and isothermal strand displacement amplification method without the requirement of a stepwise procedure, which greatly simplifies the experimental procedure and decreases the probability of contamination of samples. With its advantages, the method would be very useful to detect nucleic acids in point-of-care or field use.

  5. Validation of a multi-analyte HPLC-DAD method for determination of uric acid, creatinine, homovanillic acid, niacinamide, hippuric acid, indole-3-acetic acid and 2-methylhippuric acid in human urine.

    PubMed

    Remane, Daniela; Grunwald, Soeren; Hoeke, Henrike; Mueller, Andrea; Roeder, Stefan; von Bergen, Martin; Wissenbach, Dirk K

    2015-08-15

    During the last decades exposure sciences and epidemiological studies attracts more attention to unravel the mechanisms for the development of chronic diseases. According to this an existing HPLC-DAD method for determination of creatinine in urine samples was expended for seven analytes and validated. Creatinine, uric acid, homovanillic acid, niacinamide, hippuric acid, indole-3-acetic acid, and 2-methylhippuric acid were separated by gradient elution (formate buffer/methanol) using an Eclipse Plus C18 Rapid Resolution column (4.6mm×100mm). No interfering signals were detected in mobile phase. After injection of blank urine samples signals for the endogenous compounds but no interferences were detected. All analytes were linear in the selected calibration range and a non weighted calibration model was chosen. Bias, intra-day and inter-day precision for all analytes were below 20% for quality control (QC) low and below 10% for QC medium and high. The limits of quantification in mobile phase were in line with reported reference values but had to be adjusted in urine for homovanillic acid (45mg/L), niacinamide 58.5(mg/L), and indole-3-acetic acid (63mg/L). Comparison of creatinine data obtained by the existing method with those of the developed method showing differences from -120mg/L to +110mg/L with a mean of differences of 29.0mg/L for 50 authentic urine samples. Analyzing 50 authentic urine samples, uric acid, creatinine, hippuric acid, and 2-methylhippuric acid were detected in (nearly) all samples. However, homovanillic acid was detected in 40%, niacinamide in 4% and indole-3-acetic acid was never detected within the selected samples.

  6. DimaSense™: A Novel Nucleic Acid Detection System

    SciTech Connect

    Stadler, A.

    2011-05-18

    Recently, we developed a suite of methods for the rational design and fabrication of well-defined nanoparticle architectures, including clusters using bio-encoded nanoscale building blocks and layer-by-layer stepwise assembly on a solid support. In particular, the Nano-Assembly platform using Encoded Solid Supports (NAESS) allows for controlled interactions, purification of side products, modularity of design, and the construction of complex nanoparticle architectures. This approach offers several advantages over the current art of designing nanoparticle clusters, which include the high-yield synthesis of desired architectures, a 'plug-and-play' design allowing for the introduction of a variety of sensing modalities, and ease of scalability in high-throughput and synthesis yield. As a utility proof of concept, we implemented our unique cluster fabrication platform to design gold nanoparticle dimers which are linked via a single-stranded DNA oligonucleotide recognition motif. The design of this motif is such that binding of complementary nucleic acids results in specific, selective and rapid dimer dissociation, which can be monitored by dynamic light scattering (DLS). We demonstrated single level mismatch selectivity using this approach. The limit of detection was determined to be 1011 molecules of synthetic target RNA or DNA within 30 minutes of incubation at 33 C. This detection limit is determined by the dimer's concentration which can be probed by currently used standard DLS instruments. We also demonstrated a specific detection of target RNA in a solution containing competing 1,000-fold excess of non-complementary DNA fragments, 10% BSA, and endonucleases. Molecular diagnostic companies, RNA-based technology developers, and personalized medicine companies have applications that could benefit from using DimaSense{trademark}. The technology represents a platform which enables the simple and reasonably inexpensive design and fabrication of highly selective genetic

  7. Need for new caries detection methods

    NASA Astrophysics Data System (ADS)

    Young, Douglas A.; Featherstone, John D. B.

    1999-05-01

    Dental caries (tooth decay) continues to be a major problems for adults as well as children, even though great advances have been made in preventive methods in the last 20 years. New methods for the management of caries will work best if lesions can be detected at an early stage and chemical rather than physical intervention can take place, thereby preserving the natural tooth structure and helping the saliva to heal, or remineralize, the areas of early decay. Clinical detection of caries in the US relies on visual examination, tactile with hand held explorer, and conventional radiographs, all of which are inadequate for the occlusal (biting) surfaces of the teeth where most of the decay now occurs. The dentist often has to explore by drilling with a dental bur to confirm early decay in these areas. New method that can determine the extent and degree of subsurface lesions in these surfaces non-destructively are essential for further advances in the clinical management of dental caries. Optical methods, which exploit the differences between sound and carious enamel and dentin, show great promise for the accurate detection of these lesions. Two or three- dimensional images, which include a measure of severity will be needed.

  8. Method of Identifying a Base in a Nucleic Acid

    DOEpatents

    Fodor, Stephen P. A.; Lipshutz, Robert J.; Huang, Xiaohua

    1999-01-01

    Devices and techniques for hybridization of nucleic acids and for determining the sequence of nucleic acids. Arrays of nucleic acids are formed by techniques, preferably high resolution, light-directed techniques. Positions of hybridization of a target nucleic acid are determined by, e.g., epifluorescence microscopy. Devices and techniques are proposed to determine the sequence of a target nucleic acid more efficiently and more quickly through such synthesis and detection techniques.

  9. Improving acid-fast fluorescent staining for the detection of mycobacteria using a new nucleic acid staining approach.

    PubMed

    Ryan, Gavin J; Shapiro, Howard M; Lenaerts, Anne J

    2014-09-01

    Acid fast staining of sputum smears by microscopy remains the prevalent method for detecting Mycobacterium tuberculosis. The sensitivity of microscopy using acid fast stains requires 10(4) bacilli per ml of sputum. Although fluorescent acid fast stains, such as Auramine-O, show improved sensitivity, almost half of culture-positive TB cases are currently estimated to remain smear-negative. These current diagnosis problems provide impetus for improving staining procedures. We evaluated a novel fluorescent acid-fast staining approach using the nucleic acid-binding dye SYBR(®) Gold on mycobacterial in vitro cultures. The SYBR(®) Gold stain detected 99% of MTB in both actively replicating aerobic and non-replicating hypoxic cultures. Transmission light microscopy with Ziehl-Neelsen fuchsin, and fluorescence microscopy with Auramine-O or Auramine-rhodamine detected only 54%-86% of MTB bacilli. SYBR(®) Gold fluoresces more intensely than Auramine-O, and is highly resistant to fading. The signal to noise ratio is exceptionally high due to a >1000-fold enhanced fluorescence after binding to DNA/RNA, thereby reducing most background fluorescence. Although cost and stability of the dye may perhaps limit its clinical use at this time, these results warrant further research into more nucleic acid dye variants. In the meantime, SYBR(®) Gold staining shows great promise for use in numerous research applications.

  10. Analytical Methods for Exoplanet Imaging Detection Metrics

    NASA Astrophysics Data System (ADS)

    Garrett, Daniel; Savransky, Dmitry

    2017-01-01

    When designing or simulating exoplanet-finding missions, a selection metric must be used to choose which target stars will be observed. For direct imaging missions, the metric is a function of the planet-star separation and flux ratio as constrained by the instrument's inner and outer working angles and contrast. We present analytical methods for the calculation of two detection metrics: completeness and depth of search. While Monte Carlo methods have typically been used for determining each of these detection metrics, implementing analytical methods in simulation or early stage design yields quicker, more accurate calculations.Completeness is the probability of detecting a planet belonging to the planet population of interest. This metric requires assumptions to be made about the planet population. Probability density functions are assumed for the planetary parameters of semi-major axis, eccentricity, geometric albedo, and planetary radius. Planet-star separation and difference in brightness magnitude or contrast are written as functions of these parameters. A change of variables is performed to get a joint probability density function of planet-star separation and difference in brightness magnitude or contrast. This joint probability density function is marginalized subject to the constraints of the instrument to yield the probability of detecting a planet belonging to the population of interest.Depth of search for direct imaging is the sum of the probability of detecting a planet of given semi-major axis and planetary radius by a given instrument for a target list. This metric does not depend on assumed planet population parameter distributions. A two-dimensional grid of probabilities is generated for each star in the target list. The probability at each point in the grid is found by marginalizing a probability density function of contrast given constant values of semi-major axis and planetary radius subject to the constraints of the instrument.

  11. Research and Design of Rootkit Detection Method

    NASA Astrophysics Data System (ADS)

    Liu, Leian; Yin, Zuanxing; Shen, Yuli; Lin, Haitao; Wang, Hongjiang

    Rootkit is one of the most important issues of network communication systems, which is related to the security and privacy of Internet users. Because of the existence of the back door of the operating system, a hacker can use rootkit to attack and invade other people's computers and thus he can capture passwords and message traffic to and from these computers easily. With the development of the rootkit technology, its applications are more and more extensive and it becomes increasingly difficult to detect it. In addition, for various reasons such as trade secrets, being difficult to be developed, and so on, the rootkit detection technology information and effective tools are still relatively scarce. In this paper, based on the in-depth analysis of the rootkit detection technology, a new kind of the rootkit detection structure is designed and a new method (software), X-Anti, is proposed. Test results show that software designed based on structure proposed is much more efficient than any other rootkit detection software.

  12. [Determination of amino acids in honey by capillary electrophoresis with indirect ultraviolet detection].

    PubMed

    Zhou, Xianjing; Shi, Yanping

    2013-07-01

    A method of capillary electrophoresis with indirect ultraviolet (UV) detection was developed for the separation and determination of nine amino acids such as lysine, tryptophan, glutamic acid, etc. The effects of sodium dihydrogen phosphates concentration, pH of buffer and sample injection type and time on the reproducibility and efficiency were investigated. The optimum injection time was 5 s at 5 kPa. The optimum electrophoretic conditions were as follow: 10 mmol/L sodium dihydrogen phosphates (pH 10. 2) containing 0. 5 mmol/L cetrimonium bromide, 20 mmol/L nicotinic acid and 10% (v/v) methanol as running buffer, applied voltage of - 15 kV, detection wavelength of 220 nm. The base line separation of the nine amino acids was achieved successfully within 11 min. The lowest detection limit was 0. 3 mg/L. All of the nine analytes showed good linearities within 1. 0 - 1000 mg/L. The relative standard deviations of migration time and peak area were 0. 64% - 5. 83%. The recoveries of the eight amino acids spiked in a real sample were between 60. 00% and 118.37%. The method was applied in the determination of the amino acids in honey samples from different nectar plants and origins. Prolin, serine and aspartic acid were found in five honey samples, and tryptophan was only found in a litchi honey sample. This method can provide good reference to the evaluation of the quality and nectar origin of honey.

  13. System and method for anomaly detection

    DOEpatents

    Scherrer, Chad

    2010-06-15

    A system and method for detecting one or more anomalies in a plurality of observations is provided. In one illustrative embodiment, the observations are real-time network observations collected from a stream of network traffic. The method includes performing a discrete decomposition of the observations, and introducing derived variables to increase storage and query efficiencies. A mathematical model, such as a conditional independence model, is then generated from the formatted data. The formatted data is also used to construct frequency tables which maintain an accurate count of specific variable occurrence as indicated by the model generation process. The formatted data is then applied to the mathematical model to generate scored data. The scored data is then analyzed to detect anomalies.

  14. Surface property detection apparatus and method

    DOEpatents

    Martens, Jon S.; Ginley, David S.; Hietala, Vincent M.; Sorensen, Neil R.

    1995-01-01

    Apparatus and method for detecting, determining, and imaging surface resistance corrosion, thin film growth, and oxide formation on the surface of conductors or other electrical surface modification. The invention comprises a modified confocal resonator structure with the sample remote from the radiating mirror. Surface resistance is determined by analyzing and imaging reflected microwaves; imaging reveals anomalies due to surface impurities, non-stoichiometry, and the like, in the surface of the superconductor, conductor, dielectric, or semiconductor.

  15. Surface property detection apparatus and method

    DOEpatents

    Martens, J.S.; Ginley, D.S.; Hietala, V.M.; Sorensen, N.R.

    1995-08-08

    Apparatus and method for detecting, determining, and imaging surface resistance corrosion, thin film growth, and oxide formation on the surface of conductors or other electrical surface modification. The invention comprises a modified confocal resonator structure with the sample remote from the radiating mirror. Surface resistance is determined by analyzing and imaging reflected microwaves; imaging reveals anomalies due to surface impurities, non-stoichiometry, and the like, in the surface of the superconductor, conductor, dielectric, or semiconductor. 4 figs.

  16. Graphdiyne as a promising material for detecting amino acids

    NASA Astrophysics Data System (ADS)

    Chen, Xi; Gao, Pengfei; Guo, Lei; Zhang, Shengli

    2015-11-01

    The adsorption of glycine, glutamic acid, histidine and phenylalanine on single-layer graphdiyne/ graphene is investigated by ab initio calculations. The results show that for each amino acid molecule, the adsorption energy on graphdiyne is larger than the adsorption energy on graphene and dispersion interactions predominate in the adsorption. Molecular dynamics simulations reveal that at room temperature the amino acid molecules keep migrating and rotating on graphdiyne surface and induce fluctuation in graphdiyne bandgap. Additionally, the photon absorption spectra of graphdiyne-amino-acid systems are investigated. We uncover that the presence of amino acid molecules makes the photon absorption peaks of graphdiyne significantly depressed and shifted. Finally, quantum electronic transport properties of graphdiyne-amino-acid systems are compared with the transport properties of pure graphdiyne. We reveal that the amino acid molecules induce distinct changes in the electronic conductivity of graphdiyne. The results in this paper reveal that graphdiyne is a promising two-dimensional material for sensitively detecting amino acids and may potentially be used in biosensors.

  17. Seismic Anomaly Detection Using Symbolic Representation Methods

    NASA Astrophysics Data System (ADS)

    Christodoulou, Vyron; Bi, Yaxin; Wilkie, George; Zhao, Guoze

    2016-08-01

    In this work we investigate the use of symbolic representation methods for Anomaly Detection in different electromagnetic sequential time series datasets. An issue that is often overlooked regarding symbolic representation and its performance in Anomaly Detection is the use of a quantitative accuracy metric. Until recently only visual representations have been used to show the efficiency of an algorithm to detect anomalies. In this respect we propose an novel accuracy metric that takes into account the length of the sliding window of such symbolic representation algorithms and we present its utility. For the evaluation of the accuracy metric, HOT-SAX is used, a method that aggregates data points by use of sliding windows. A HOT-SAX variant, with the use of overlapping windows, is also introduced that achieves better results based on the newly defined accuracy metric. Both methods are evaluated on ten different benchmark datasets and based on the empirical evidence we use Earth's geomagnetic data gathered by the SWARM satellites and terrestrial sources around the epicenter of two seismic events in the Yunnan region of China.

  18. Detection Method of TOXOPLASMA GONDII Tachyzoites

    NASA Astrophysics Data System (ADS)

    Eassa, Souzan; Bose, Chhanda; Alusta, Pierre; Tarasenko, Olga

    2011-06-01

    Tachyzoites are considered to be the most important stage of Toxoplasma gondii which causes toxoplasmosis. T. gondii is, an obligate intracellular parasite which infects a wide range of cells. The present study was designed to develop a method for an early detection of T. gondii tachyzoites. The method comprised of a binding assay which was analyzed using principal component and cluster analysis. Our data showed that glycoconjugates GC1, GC2, GC3 and GC10 exhibit a significantly higher binding affinity for T. gondii tachyzoites as compared to controls (T. gondii only, PAA only, GC 1, 2, 3, and 10 only).

  19. Improved method for detection of starch hydrolysis

    SciTech Connect

    Ohawale, M.R.; Wilson, J.J.; Khachatourians, G.G.; Ingledew, W.M.

    1982-09-01

    A new starch hydrolysis detection method which does not rely on iodine staining or the use of color-complexed starch is described. A linear relationship was obtained with agar-starch plates when net clearing zones around colonies of yeasts were plotted against enzyme levels (semilogarithm scale) produced by the same yeast strains in liquid medium. A similar relationship between starch clearing zones and alpha-amylase levels from three different sources was observed. These observations suggest that the method is useful in mutant isolations, strain improvement programs, and the prediction of alpha-amylase activities in culture filtrates or column effluents. (Refs. 18).

  20. Optimization of Quantitative PCR Methods for Enteropathogen Detection.

    PubMed

    Liu, Jie; Gratz, Jean; Amour, Caroline; Nshama, Rosemary; Walongo, Thomas; Maro, Athanasia; Mduma, Esto; Platts-Mills, James; Boisen, Nadia; Nataro, James; Haverstick, Doris M; Kabir, Furqan; Lertsethtakarn, Paphavee; Silapong, Sasikorn; Jeamwattanalert, Pimmada; Bodhidatta, Ladaporn; Mason, Carl; Begum, Sharmin; Haque, Rashidul; Praharaj, Ira; Kang, Gagandeep; Houpt, Eric R

    2016-01-01

    Detection and quantification of enteropathogens in stool specimens is useful for diagnosing the cause of diarrhea but is technically challenging. Here we evaluate several important determinants of quantification: specimen collection, nucleic acid extraction, and extraction and amplification efficiency. First, we evaluate the molecular detection and quantification of pathogens in rectal swabs versus stool, using paired flocked rectal swabs and whole stool collected from 129 children hospitalized with diarrhea in Tanzania. Swabs generally yielded a higher quantification cycle (Cq) (average 29.7, standard deviation 3.5 vs. 25.3 ± 2.9 from stool, P<0.001) but were still able to detect 80% of pathogens with a Cq < 30 in stool. Second, a simplified total nucleic acid (TNA) extraction procedure was compared to separate DNA and RNA extractions and showed 92% (318/344) sensitivity and 98% (951/968) specificity, with no difference in Cq value for the positive results (ΔCq(DNA+RNA-TNA) = -0.01 ± 1.17, P = 0.972, N = 318). Third, we devised a quantification scheme that adjusts pathogen quantity to the specimen's extraction and amplification efficiency, and show that this better estimates the quantity of spiked specimens than the raw target Cq. In sum, these methods for enteropathogen quantification, stool sample collection, and nucleic acid extraction will be useful for laboratories studying enteric disease.

  1. Optimization of Quantitative PCR Methods for Enteropathogen Detection

    PubMed Central

    Liu, Jie; Gratz, Jean; Amour, Caroline; Nshama, Rosemary; Walongo, Thomas; Maro, Athanasia; Mduma, Esto; Platts-Mills, James; Boisen, Nadia; Nataro, James; Haverstick, Doris M.; Kabir, Furqan; Lertsethtakarn, Paphavee; Silapong, Sasikorn; Jeamwattanalert, Pimmada; Bodhidatta, Ladaporn; Mason, Carl; Begum, Sharmin; Haque, Rashidul; Praharaj, Ira; Kang, Gagandeep; Houpt, Eric R.

    2016-01-01

    Detection and quantification of enteropathogens in stool specimens is useful for diagnosing the cause of diarrhea but is technically challenging. Here we evaluate several important determinants of quantification: specimen collection, nucleic acid extraction, and extraction and amplification efficiency. First, we evaluate the molecular detection and quantification of pathogens in rectal swabs versus stool, using paired flocked rectal swabs and whole stool collected from 129 children hospitalized with diarrhea in Tanzania. Swabs generally yielded a higher quantification cycle (Cq) (average 29.7, standard deviation 3.5 vs. 25.3 ± 2.9 from stool, P<0.001) but were still able to detect 80% of pathogens with a Cq < 30 in stool. Second, a simplified total nucleic acid (TNA) extraction procedure was compared to separate DNA and RNA extractions and showed 92% (318/344) sensitivity and 98% (951/968) specificity, with no difference in Cq value for the positive results (ΔCq(DNA+RNA-TNA) = -0.01 ± 1.17, P = 0.972, N = 318). Third, we devised a quantification scheme that adjusts pathogen quantity to the specimen’s extraction and amplification efficiency, and show that this better estimates the quantity of spiked specimens than the raw target Cq. In sum, these methods for enteropathogen quantification, stool sample collection, and nucleic acid extraction will be useful for laboratories studying enteric disease. PMID:27336160

  2. A graphene-based electrochemical competitive immunosensor for the sensitive detection of okadaic acid in shellfish

    NASA Astrophysics Data System (ADS)

    Eissa, Shimaa; Zourob, Mohammed

    2012-11-01

    A novel graphene-based voltammetric immunosensor for sensitive detection of okadaic acid (OA) was developed. A simple and efficient electrografting method was utilized to functionalize graphene-modified screen-printed carbon electrodes (GSPE) by the electrochemical reduction of in situ generated 4-carboxyphenyl diazonium salt in acidic aqueous solution. Next, the okadaic acid antibody was covalently immobilized on the carboxyphenyl modified graphene electrodes via carbodiimide chemistry. Square wave voltammetry (SWV) was used to investigate the stepwise assembly of the immunosensor. A competitive assay between OA and a fixed concentration of okadaic acid-ovalbumin conjugate (OA-OVA) for the immobilized antibodies was employed for the detection of okadaic acid. The decrease of the [Fe(CN)6]3-/4- reduction peak current in the square wave voltammetry for various concentrations of okadaic acid was used for establishing the calibration curve. A linear relationship between the SWV peak current difference and OA concentration was obtained up to ~5000 ng L-1. The developed immunosensor allowed a detection limit of 19 ng L-1 of OA in PBS buffer. The matrix effect studied with spiked shellfish tissue extracts showed a good percentage of recovery and the method was also validated with certified reference mussel samples.A novel graphene-based voltammetric immunosensor for sensitive detection of okadaic acid (OA) was developed. A simple and efficient electrografting method was utilized to functionalize graphene-modified screen-printed carbon electrodes (GSPE) by the electrochemical reduction of in situ generated 4-carboxyphenyl diazonium salt in acidic aqueous solution. Next, the okadaic acid antibody was covalently immobilized on the carboxyphenyl modified graphene electrodes via carbodiimide chemistry. Square wave voltammetry (SWV) was used to investigate the stepwise assembly of the immunosensor. A competitive assay between OA and a fixed concentration of okadaic acid

  3. Kit for detecting nucleic acid sequences using competitive hybridization probes

    DOEpatents

    Lucas, Joe N.; Straume, Tore; Bogen, Kenneth T.

    2001-01-01

    A kit is provided for detecting a target nucleic acid sequence in a sample, the kit comprising: a first hybridization probe which includes a nucleic acid sequence that is sufficiently complementary to selectively hybridize to a first portion of the target sequence, the first hybridization probe including a first complexing agent for forming a binding pair with a second complexing agent; and a second hybridization probe which includes a nucleic acid sequence that is sufficiently complementary to selectively hybridize to a second portion of the target sequence to which the first hybridization probe does not selectively hybridize, the second hybridization probe including a detectable marker; a third hybridization probe which includes a nucleic acid sequence that is sufficiently complementary to selectively hybridize to a first portion of the target sequence, the third hybridization probe including the same detectable marker as the second hybridization probe; and a fourth hybridization probe which includes a nucleic acid sequence that is sufficiently complementary to selectively hybridize to a second portion of the target sequence to which the third hybridization probe does not selectively hybridize, the fourth hybridization probe including the first complexing agent for forming a binding pair with the second complexing agent; wherein the first and second hybridization probes are capable of simultaneously hybridizing to the target sequence and the third and fourth hybridization probes are capable of simultaneously hybridizing to the target sequence, the detectable marker is not present on the first or fourth hybridization probes and the first, second, third, and fourth hybridization probes each include a competitive nucleic acid sequence which is sufficiently complementary to a third portion of the target sequence that the competitive sequences of the first, second, third, and fourth hybridization probes compete with each other to hybridize to the third portion of the

  4. Fast hybridization solution for the detection of immobilized nucleic acids.

    PubMed

    Yang, T T; Kain, S R

    1995-03-01

    We have developed a fast hybridization solution, termed ExpressHyb, for the rapid and sensitive detection of nucleic acids immobilized on membrane supports. This solution reduces typical hybridization times of 12-24 h to as little as 1 h while simultaneously increasing the sensitivity of detection in many applications. Using ExpressHyb, human beta-actin mRNA was detected on a human multiple tissue Northern (MTN) blot following a 30-min hybridization, with optimal detection occurring with a 1-h hybridization interval. The moderately abundant human glyceraldehyde-3-phosphate dehydrogenase (G3PDH) mRNA was detected using similar hybridization conditions and yielded improved signal-to-background characteristics relative to overnight hybridizations in conventional solutions. ExpressHyb can be used with either 32P- or digoxigenin-labeled probes and works effectively with both cDNA and oligonucleotide probes. For non-isotopic detection in particular, ExpressHyb reduces the nonspecific background commonly encountered with this technique. In cDNA library screening, ExpressHyb was found to both reduce the time required for effective hybridizations and to increase the number of positive colonies obtained relative to conventional overnight procedures. Taken together, these results illustrate the broad capability of ExpressHyb Hybridization Solution to improve nucleic acid detection in a variety of important techniques.

  5. Determination of L- Ascorbic Acid in Plasma by Voltammetric Method

    PubMed Central

    Behfar, Abdol Azim; Sadeghi, Nafiseh; Jannat, Behrooz; Oveisi, Mohammad Reza

    2010-01-01

    Voltammetric techniques have been considered as important methods among the analytical techniques used for the identification and determination of trace concentrations of many biological molecules such as L-ascorbic acid (AA). L-ascorbic acid is an electro-active molecule, though it is difficult to determine its value directly with a majority of electrodes made of carbon and transition metals, because of electrode surface problems. The present study is based on I-E curves for AA analysis at various pH. Furthermore, the effects of the presence of other electro-active substances; such as copper, as well as the effect of the sweep rate of potential will be studied. The present study is based on analysis of the current-voltage curves for L-ascorbic acid at varying pH and sweep rate scan values. An analysis was also carried out to measure the influence of the concentration of some electro active species. The peak height of the first oxidation wave is used for L-ascorbic acid assay. L-ascorbic acid was determined in aqueous media by linear-scan voltammetry on a gold electrode; ranging between (1-175 μg/mL). In biologic samples, for elimination of uric acid or some sugars and effects, a significant interference of copper ions whose presence reduces the height of the L-ascorbic acid oxidation peak was used. The optimum pH and sweep rate were 3.2 and 7500mV/s, respectively. Under these conditions, the detection limit of the method was 0.3 μg/mL. Repeatability of the method based on relative standard deviation (RSD) 50, 10 and 1 μg/mL concentrations was 0.83, 2.1 and 10.3%, respectively. The calibration curve was linear over the range 1-175μg/mL (r2 = 0.9977, p < 0.001). The advantage of this method lies in the fact that the use of copper eliminates the interference of different substances such as uric acid. PMID:24363717

  6. Method of increasing conversion of a fatty acid to its corresponding dicarboxylic acid

    DOEpatents

    Craft, David L.; Wilson, C. Ron; Eirich, Dudley; Zhang, Yeyan

    2004-09-14

    A nucleic acid sequence including a CYP promoter operably linked to nucleic acid encoding a heterologous protein is provided to increase transcription of the nucleic acid. Expression vectors and host cells containing the nucleic acid sequence are also provided. The methods and compositions described herein are especially useful in the production of polycarboxylic acids by yeast cells.

  7. Ultrasensitive nucleic acid sequence detection by single-molecule electrophoresis

    SciTech Connect

    Castro, A; Shera, E.B.

    1996-09-01

    This is the final report of a one-year laboratory-directed research and development project at Los Alamos National Laboratory. There has been considerable interest in the development of very sensitive clinical diagnostic techniques over the last few years. Many pathogenic agents are often present in extremely small concentrations in clinical samples, especially at the initial stages of infection, making their detection very difficult. This project sought to develop a new technique for the detection and accurate quantification of specific bacterial and viral nucleic acid sequences in clinical samples. The scheme involved the use of novel hybridization probes for the detection of nucleic acids combined with our recently developed technique of single-molecule electrophoresis. This project is directly relevant to the DOE`s Defense Programs strategic directions in the area of biological warfare counter-proliferation.

  8. Waterborne Pathogens: Detection Methods and Challenges

    PubMed Central

    Ramírez-Castillo, Flor Yazmín; Loera-Muro, Abraham; Jacques, Mario; Garneau, Philippe; Avelar-González, Francisco Javier; Harel, Josée; Guerrero-Barrera, Alma Lilián

    2015-01-01

    Waterborne pathogens and related diseases are a major public health concern worldwide, not only by the morbidity and mortality that they cause, but by the high cost that represents their prevention and treatment. These diseases are directly related to environmental deterioration and pollution. Despite the continued efforts to maintain water safety, waterborne outbreaks are still reported globally. Proper assessment of pathogens on water and water quality monitoring are key factors for decision-making regarding water distribution systems’ infrastructure, the choice of best water treatment and prevention waterborne outbreaks. Powerful, sensitive and reproducible diagnostic tools are developed to monitor pathogen contamination in water and be able to detect not only cultivable pathogens but also to detect the occurrence of viable but non-culturable microorganisms as well as the presence of pathogens on biofilms. Quantitative microbial risk assessment (QMRA) is a helpful tool to evaluate the scenarios for pathogen contamination that involve surveillance, detection methods, analysis and decision-making. This review aims to present a research outlook on waterborne outbreaks that have occurred in recent years. This review also focuses in the main molecular techniques for detection of waterborne pathogens and the use of QMRA approach to protect public health. PMID:26011827

  9. Method for nucleic acid hybridization using single-stranded DNA binding protein

    DOEpatents

    Tabor, Stanley; Richardson, Charles C.

    1996-01-01

    Method of nucleic acid hybridization for detecting the presence of a specific nucleic acid sequence in a population of different nucleic acid sequences using a nucleic acid probe. The nucleic acid probe hybridizes with the specific nucleic acid sequence but not with other nucleic acid sequences in the population. The method includes contacting a sample (potentially including the nucleic acid sequence) with the nucleic acid probe under hybridizing conditions in the presence of a single-stranded DNA binding protein provided in an amount which stimulates renaturation of a dilute solution (i.e., one in which the t.sub.1/2 of renaturation is longer than 3 weeks) of single-stranded DNA greater than 500 fold (i.e., to a t.sub.1/2 less than 60 min, preferably less than 5 min, and most preferably about 1 min.) in the absence of nucleotide triphosphates.

  10. Current methods for detecting ethylene in plants

    PubMed Central

    Cristescu, Simona M.; Mandon, Julien; Arslanov, Denis; De Pessemier, Jérôme; Hermans, Christian; Harren, Frans J. M.

    2013-01-01

    Background In view of ethylene's critical developmental and physiological roles the gaseous hormone remains an active research topic for plant biologists. Progress has been made to understand the ethylene biosynthesis pathway and the mechanisms of perception and action. Still numerous questions need to be answered and findings to be validated. Monitoring gas production will very often complete the picture of any ethylene research topic. Therefore the search for suitable ethylene measuring methods for various plant samples either in the field, greenhouses, laboratories or storage facilities is strongly motivated. Scope This review presents an update of the current methods for ethylene monitoring in plants. It focuses on the three most-used methods – gas chromatography detection, electrochemical sensing and optical detection – and compares them in terms of sensitivity, selectivity, time response and price. Guidelines are provided for proper selection and application of the described sensor methodologies and some specific applications are illustrated of laser-based detector for monitoring ethylene given off by Arabidopsis thaliana upon various nutritional treatments. Conclusions Each method has its advantages and limitations. The choice for the suitable ethylene sensor needs careful consideration and is driven by the requirements for a specific application. PMID:23243188

  11. A new method for FMRI activation detection

    NASA Astrophysics Data System (ADS)

    Wei, Jianing; Talavage, Thomas M.; Pollak, Ilya

    2009-02-01

    The objective of fMRI data analysis is to detect the region of the brain that gets activated in response to a specific stimulus presented to the subject. We develop a new algorithm for activation detection in event-related fMRI data. We utilize a forward model for fMRI data acquisition which explicitly incorporates physiological noise, scanner noise and the spatial blurring introduced by the scanner. After slice-by-slice image restoration procedure that independently restores each data slice corresponding to each time index, we estimate the parameters of the hemodynamic response function (HRF) model for each pixel of the restored data. In order to enforce spatial regularity in our estimates, we model the prior distribution of the HRF parameters as a generalized Gaussian Markov random field (GGMRF) model. We develop an algorithm to compute the maximum a posteriori (MAP) estimates of the parameters. We then threshold the amplitude parameters to obtain the final activation map. We illustrate our algorithm by comparing it with the widely used general linear model (GLM) method. In synthetic data experiments, under the same probability of false alarm, the probability of correct detection for our method is up to 15% higher than GLM. In real data experiments, through anatomical analysis and benchmark testing using block paradigm results, we demonstrate that our algorithm produces fewer false alarms than GLM.

  12. Design of acid-lead battery stage-of-charge detection system based on refractive index detection technology

    NASA Astrophysics Data System (ADS)

    Chen, Junyao; Yang, Kecheng; Xia, Min; Li, Lei; Zeng, Xianjiang

    2015-10-01

    Based on optical total reflection critical Angle method, we have designed a refractive index measurement system. It adopted a divergent light source and a CCD camera as the occurrence and receiver of the signal. The divergent light source sent out a bunch of tapered beam, exposure to the interface of optical medium and sulfuric acid solution. Light intensity reflected from the interface could be detected by the CCD camera and then sent to the embedded system. In the DSP embedded system, we could obtain the critical edge position through the light intensity distribution curve and converted it to critical angle. Through experiment, we concluded the relation between liquid refractive index and the critical angle edge position. In this system, the detecting precision of the refractive index of sulfuric acid solution reached 10-4. Finally, through the conversion of the refractive index and density, we achieved high accuracy online measurement of electrolyte density in lead-acid battery.

  13. Halocarbon refrigerant detection methods. Final report

    SciTech Connect

    Tapscott, R.E.; Sohn, C.W.

    1996-01-01

    The Montreal Protocol and the U.S. Clean Air Act limit the production of ozone-depleting substances, including many refrigerants. Three options for cost-effectively phasing out these refrigerants from Army installations are: (1) refrigerant containment, (2) retrofit conversion to accommodate alternative refrigerant, and (3) replacement with cooling systems using alternative refrigerant. This report contributes to the first option by identifying and assessing methods to detect chlorofluorocarbon (CFC), hydrochlorofluorocarbon (HCFC) and hydrofluorocarbon (HFC) refrigerants that leak from air-conditioning and refrigeration systems. As background, the report describes the relevant sections of the Montreal Protocol and the Clean Air Act, and gives an overview of refrigerants. This is followed by a description of the technologies used in refrigerant leak detection, and a survey of detector types available and their price ranges. Appendixes provide an extensive list of detector products and their specifications, plus manufacturer addresses and phone numbers.

  14. Correlation fluorescence method of amine detection

    NASA Astrophysics Data System (ADS)

    Myslitsky, Valentin F.; Tkachuk, Svetlana S.; Rudeichuk, Volodimir M.; Strinadko, Miroslav T.; Slyotov, Mikhail M.; Strinadko, Marina M.

    1997-12-01

    The amines fluorescence spectra stimulated by UV laser radiation are investigated in this paper. The fluorescence is stimulated by the coherent laser beam with the wavelength 0.337 micrometers . At the sufficient energy of laser stimulation the narrow peaks of the fluorescence spectra are detected besides the wide maximum. The relationship between the fluorescence intensity and the concentration of amines solutions are investigated. The fluorescence intensity temporal dependence on wavelength 0.363 micrometers of the norepinephrine solution preliminarily radiated by UV laser with wavelength 0.337 micrometers was found. The computer stimulated and experimental investigations of adrenaline and norepinephrine mixtures fluorescence spectra were done. The correlation fluorescent method of amines detection is proposed.

  15. ULTRASONIC FLAW DETECTION METHOD AND MEANS

    DOEpatents

    Worlton, D.C.

    1961-08-15

    A method of detecting subsurface flaws in an object using ultrasonic waves is described. An ultnasonic wave of predetermined velocity and frequency is transmitted to engage the surface of the object at a predetermined angle of inci dence thereto. The incident angle of the wave to the surface is determined with respect to phase velocity, incident wave velocity, incident wave frequency, and the estimated depth of the flaw so that Lamb waves of a particular type and mode are induced only in the portion of the object between the flaw and the surface. These Lamb waves are then detected as they leave the object at an angle of exit equal to the angle of incidence. No waves wlll be generated in the object and hence received if no flaw exists beneath the surface. (AEC)

  16. Bayesian Methods for Radiation Detection and Dosimetry

    SciTech Connect

    Peter G. Groer

    2002-09-29

    We performed work in three areas: radiation detection, external and internal radiation dosimetry. In radiation detection we developed Bayesian techniques to estimate the net activity of high and low activity radioactive samples. These techniques have the advantage that the remaining uncertainty about the net activity is described by probability densities. Graphs of the densities show the uncertainty in pictorial form. Figure 1 below demonstrates this point. We applied stochastic processes for a method to obtain Bayesian estimates of 222Rn-daughter products from observed counting rates. In external radiation dosimetry we studied and developed Bayesian methods to estimate radiation doses to an individual with radiation induced chromosome aberrations. We analyzed chromosome aberrations after exposure to gammas and neutrons and developed a method for dose-estimation after criticality accidents. The research in internal radiation dosimetry focused on parameter estimation for compartmental models from observed compartmental activities. From the estimated probability densities of the model parameters we were able to derive the densities for compartmental activities for a two compartment catenary model at different times. We also calculated the average activities and their standard deviation for a simple two compartment model.

  17. Method and apparatus for detecting neutrons

    DOEpatents

    Perkins, Richard W.; Reeder, Paul L.; Wogman, Ned A.; Warner, Ray A.; Brite, Daniel W.; Richey, Wayne C.; Goldman, Don S.

    1997-01-01

    The instant invention is a method for making and using an apparatus for detecting neutrons. Scintillating optical fibers are fabricated by melting SiO.sub.2 with a thermal neutron capturing substance and a scintillating material in a reducing atmosphere. The melt is then drawn into fibers in an anoxic atmosphere. The fibers may then be coated and used directly in a neutron detection apparatus, or assembled into a geometrical array in a second, hydrogen-rich, scintillating material such as a polymer. Photons generated by interaction with thermal neutrons are trapped within the coated fibers and are directed to photoelectric converters. A measurable electronic signal is generated for each thermal neutron interaction within the fiber. These electronic signals are then manipulated, stored, and interpreted by normal methods to infer the quality and quantity of incident radiation. When the fibers are arranged in an array within a second scintillating material, photons generated by kinetic neutrons interacting with the second scintillating material and photons generated by thermal neutron capture within the fiber can both be directed to photoelectric converters. These electronic signals are then manipulated, stored, and interpreted by normal methods to infer the quality and quantity of incident radiation.

  18. Method and apparatus for detecting neutrons

    DOEpatents

    Perkins, R.W.; Reeder, P.L.; Wogman, N.A.; Warner, R.A.; Brite, D.W.; Richey, W.C.; Goldman, D.S.

    1997-10-21

    The instant invention is a method for making and using an apparatus for detecting neutrons. Scintillating optical fibers are fabricated by melting SiO{sub 2} with a thermal neutron capturing substance and a scintillating material in a reducing atmosphere. The melt is then drawn into fibers in an anoxic atmosphere. The fibers may then be coated and used directly in a neutron detection apparatus, or assembled into a geometrical array in a second, hydrogen-rich, scintillating material such as a polymer. Photons generated by interaction with thermal neutrons are trapped within the coated fibers and are directed to photoelectric converters. A measurable electronic signal is generated for each thermal neutron interaction within the fiber. These electronic signals are then manipulated, stored, and interpreted by normal methods to infer the quality and quantity of incident radiation. When the fibers are arranged in an array within a second scintillating material, photons generated by kinetic neutrons interacting with the second scintillating material and photons generated by thermal neutron capture within the fiber can both be directed to photoelectric converters. These electronic signals are then manipulated, stored, and interpreted by normal methods to infer the quality and quantity of incident radiation. 5 figs.

  19. Detection and isolation of circulating tumor cells: principles and methods.

    PubMed

    Esmaeilsabzali, Hadi; Beischlag, Timothy V; Cox, Michael E; Parameswaran, Ash M; Park, Edward J

    2013-11-15

    Efforts to improve the clinical management of several cancers include finding better methods for the quantitative and qualitative analysis of circulating tumor cells (CTCs). However, detection and isolation of CTCs from the blood circulation is not a trivial task given their scarcity and the lack of reliable markers to identify these cells. With a variety of emerging technologies, a thorough review of the exploited principles and techniques as well as the trends observed in the development of these technologies can assist researchers to recognize the potential improvements and alternative approaches. To help better understand the related biological concepts, a simplified framework explaining cancer formation and its spread to other organs as well as how CTCs contribute to this process has been presented first. Then, based on their basic working-principles, the existing methods for detection and isolation of CTCs have been classified and reviewed as nucleic acid-based, physical properties-based and antibody-based methods. The review of literature suggests that antibody-based methods, particularly in conjunction with a microfluidic lab-on-a-chip setting, offer the highest overall performance for detection and isolation of CTCs. Further biological and engineering-related research is required to improve the existing methods. These include finding more specific markers for CTCs as well as enhancing the throughput, sensitivity, and analytic functionality of current devices.

  20. Method and apparatus for detecting explosives

    DOEpatents

    Moore, David Steven

    2011-05-10

    A method and apparatus is provided for detecting explosives by thermal imaging. The explosive material is subjected to a high energy wave which can be either a sound wave or an electromagnetic wave which will initiate a chemical reaction in the explosive material which chemical reaction will produce heat. The heat is then sensed by a thermal imaging device which will provide a signal to a computing device which will alert a user of the apparatus to the possibility of an explosive device being present.

  1. Systems and methods for detecting and processing

    DOEpatents

    Johnson, Michael M.; Yoshimura, Ann S.

    2006-03-28

    Embodiments of the present invention provides systems and method for detecting. Sensing modules are provided in communication with one or more detectors. In some embodiments, detectors are provided that are sensitive to chemical, biological, or radiological agents. Embodiments of sensing modules include processing capabilities to analyze, perform computations on, and/or run models to predict or interpret data received from one or more detectors. Embodiments of sensing modules form various network configurations with one another and/or with one or more data aggregation devices. Some embodiments of sensing modules include power management functionalities.

  2. Visual detection of nucleic acids based on Mie scattering and the magnetophoretic effect.

    PubMed

    Zhao, Zichen; Chen, Shan; Ho, John Kin Lim; Chieng, Ching-Chang; Chen, Ting-Hsuan

    2015-12-07

    Visual detection of nucleic acid biomarkers is a simple and convenient approach to point-of-care applications. However, issues of sensitivity and the handling of complex bio-fluids have posed challenges. Here we report on a visual method detecting nucleic acids using Mie scattering of polystyrene microparticles and the magnetophoretic effect. Magnetic microparticles (MMPs) and polystyrene microparticles (PMPs) were surface-functionalised with oligonucleotide probes, which can hybridise with target oligonucleotides in juxtaposition and lead to the formation of MMPs-targets-PMPs sandwich structures. Using an externally applied magnetic field, the magnetophoretic effect attracts the sandwich structure to the sidewall, which reduces the suspended PMPs and leads to a change in the light transmission via the Mie scattering. Based on the high extinction coefficient of the Mie scattering (∼3 orders of magnitude greater than that of the commonly used gold nanoparticles), our results showed the limit of detection to be 4 pM using a UV-Vis spectrometer or 10 pM by direct visual inspection. Meanwhile, we also demonstrated that this method is compatible with multiplex assays and detection in complex bio-fluids, such as whole blood or a pool of nucleic acids, without purification in advance. With a simplified operation procedure, low instrumentation requirement, high sensitivity and compatibility with complex bio-fluids, this method provides an ideal solution for visual detection of nucleic acids in resource-limited settings.

  3. DNA tetrahedron and star trigon nanostructures for target recycling detection of nucleic acid.

    PubMed

    Li, Yueran; Chen, Xifeng; Wang, Bidou; Liu, Guangxing; Tang, Yuguo; Miao, Peng

    2016-06-07

    Human immunodeficiency virus (HIV) is a retrovirus which attacks the human body's immune system and further leads to acquired immunodeficiency syndrome (AIDS). Nucleic acid detection is of great importance in the medical diagnosis of such diseases. Herein, we develop a simple and enzyme-free electrochemical method for the target recycling detection of nuclei acid. DNA tetrahedron and star trigon nanostructures are designed and constructed on the electrode interface for target capture and signal enrichment. This strategy is convenient and sensitive, with a limit of detection as low as 1 fM, and can also successfully distinguish single-base mismatched DNA. Therefore, the proposed method has a promising potential application for HIV DNA detection.

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

    PubMed Central

    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

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

    PubMed

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

    2014-01-01

    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.

  6. Traumatic Brain Injury Detection Using Electrophysiological Methods

    PubMed Central

    Rapp, Paul E.; Keyser, David O.; Albano, Alfonso; Hernandez, Rene; Gibson, Douglas B.; Zambon, Robert A.; Hairston, W. David; Hughes, John D.; Krystal, Andrew; Nichols, Andrew S.

    2015-01-01

    Measuring neuronal activity with electrophysiological methods may be useful in detecting neurological dysfunctions, such as mild traumatic brain injury (mTBI). This approach may be particularly valuable for rapid detection in at-risk populations including military service members and athletes. Electrophysiological methods, such as quantitative electroencephalography (qEEG) and recording event-related potentials (ERPs) may be promising; however, the field is nascent and significant controversy exists on the efficacy and accuracy of the approaches as diagnostic tools. For example, the specific measures derived from an electroencephalogram (EEG) that are most suitable as markers of dysfunction have not been clearly established. A study was conducted to summarize and evaluate the statistical rigor of evidence on the overall utility of qEEG as an mTBI detection tool. The analysis evaluated qEEG measures/parameters that may be most suitable as fieldable diagnostic tools, identified other types of EEG measures and analysis methods of promise, recommended specific measures and analysis methods for further development as mTBI detection tools, identified research gaps in the field, and recommended future research and development thrust areas. The qEEG study group formed the following conclusions: (1) Individual qEEG measures provide limited diagnostic utility for mTBI. However, many measures can be important features of qEEG discriminant functions, which do show significant promise as mTBI detection tools. (2) ERPs offer utility in mTBI detection. In fact, evidence indicates that ERPs can identify abnormalities in cases where EEGs alone are non-disclosing. (3) The standard mathematical procedures used in the characterization of mTBI EEGs should be expanded to incorporate newer methods of analysis including non-linear dynamical analysis, complexity measures, analysis of causal interactions, graph theory, and information dynamics. (4) Reports of high specificity in q

  7. Radiation sensitive area detection device and method

    NASA Technical Reports Server (NTRS)

    Carter, Daniel C. (Inventor); Hecht, Diana L. (Inventor); Witherow, William K. (Inventor)

    1991-01-01

    A radiation sensitive area detection device for use in conjunction with an X ray, ultraviolet or other radiation source is provided which comprises a phosphor containing film which releases a stored diffraction pattern image in response to incoming light or other electromagnetic wave. A light source such as a helium-neon laser, an optical fiber capable of directing light from the laser source onto the phosphor film and also capable of channelling the fluoresced light from the phosphor film to an integrating sphere which directs the light to a signal processing means including a light receiving means such as a photomultiplier tube. The signal processing means allows translation of the fluoresced light in order to detect the original pattern caused by the diffraction of the radiation by the original sample. The optical fiber is retained directly in front of the phosphor screen by a thin metal holder which moves up and down across the phosphor screen and which features a replaceable pinhole which allows easy adjustment of the resolution of the light projected onto the phosphor film. The device produces near real time images with high spatial resolution and without the distortion that accompanies prior art devices employing photomultiplier tubes. A method is also provided for carrying out radiation area detection using the device of the invention.

  8. Molecular detection methods of human papillomavirus (HPV).

    PubMed

    Zaravinos, Apostolos; Mammas, Ioannis N; Sourvinos, George; Spandidos, Demetrios A

    2009-01-01

    Human papillomavirus (HPV) testing can identify women at risk of cervical cancer. Currently, molecular detection methods are the gold standard for identification of HPV. The three categories of molecular assays that are available are based on the detection of HPV DNA and include (1) non-amplified hybridization assays, such as Southern transfer hybridization (STH), dot blot hybridization (DB) and in situ hybridization (ISH); (2) signal amplified hybridization assays, such as hybrid capture assays (HC2); (3) target amplification assays, such as polymerase chain reaction (PCR) and in situ PCR. STH requires large amounts of DNA, is laborious and not reproducible, while ISH has only moderate sensitivity for HPV. The sensitivity of the HC2 assay is similar to that of PCR-based assays, with high sensitivity being achieved by signal rather than target amplification. PCR-based detection is both highly sensitive and specific. Since PCR can be performed on very small amounts of DNA, it is ideal for use on specimens with low DNA content. In the future, with the advance of technology, viral DNA extraction and amplification systems will become more rapid, more sensitive, and more automated.

  9. Optical biosensor based on liquid crystal droplets for detection of cholic acid

    NASA Astrophysics Data System (ADS)

    Niu, Xiaofang; Luo, Dan; Chen, Rui; Wang, Fei; Sun, Xiaowei; Dai, Haitao

    2016-12-01

    A highly sensitive cholic acid biosensor based on 4-cyano-4‧-penthlbiphenyl (5CB) Liquid crystal droplets in phosphate buffer saline solution was reported. A radial-to-bipolar transition of 5CB droplet would be triggered during competitive reaction of CA at the sodium dodecyl sulfate surfactant-laden 5CB droplet surface. Our liquid crystal droplet sensor is a low-cost, simple and fast method for CA detection. The detection limit (5 μM) of our method is 2.4 times lower than previously report by using liquid crystal film to detection of CA.

  10. Lagrangian based methods for coherent structure detection

    SciTech Connect

    Allshouse, Michael R.; Peacock, Thomas

    2015-09-15

    There has been a proliferation in the development of Lagrangian analytical methods for detecting coherent structures in fluid flow transport, yielding a variety of qualitatively different approaches. We present a review of four approaches and demonstrate the utility of these methods via their application to the same sample analytic model, the canonical double-gyre flow, highlighting the pros and cons of each approach. Two of the methods, the geometric and probabilistic approaches, are well established and require velocity field data over the time interval of interest to identify particularly important material lines and surfaces, and influential regions, respectively. The other two approaches, implementing tools from cluster and braid theory, seek coherent structures based on limited trajectory data, attempting to partition the flow transport into distinct regions. All four of these approaches share the common trait that they are objective methods, meaning that their results do not depend on the frame of reference used. For each method, we also present a number of example applications ranging from blood flow and chemical reactions to ocean and atmospheric flows.

  11. Platelet antibody: review of detection methods

    SciTech Connect

    Schwartz, K.A.

    1988-10-01

    The driving force behind development of in vitro methods for platelet antibodies is identification of plasma factors causing platelet destruction. Early methods relied on measurement of platelet activation. Current methods are more specific and use a purified antibody against immunoglobulin or complement, which is usually labeled with /sup 125/I or tagged with an enzyme or fluorescein. Comparisons of quantitation of platelet-associated IgG show wide variability between different methods. The disparate results can be related both to differences in binding of secondary antibodies to immunoglobulin in solution compared to immunoglobulins attached to platelets and to the improper assumption that the binding ratio between the secondary detecting and primary antiplatelet antibody is one. Most assays can 1) identify neonatal isoimmune thrombocytopenia and posttransfusion purpura, 2) help to differentiate between immune and nonimmune thrombocytopenias, 3) help to sort out the offending drug when drug-induced thrombocytopenia is suspected, and 4) identify platelet alloantibodies and potential platelet donors via a cross match assay for refractory patients. However, the advantages of quantitative assays over qualitative methods with respect to predictions of patients clinical course and response to different treatments remain to be investigated. 61 references.

  12. Method and system for detecting explosives

    DOEpatents

    Reber, Edward L.; Jewell, James K.; Rohde, Kenneth W.; Seabury, Edward H.; Blackwood, Larry G.; Edwards, Andrew J.; Derr, Kurt W.

    2009-03-10

    A method of detecting explosives in a vehicle includes providing a first rack on one side of the vehicle, the rack including a neutron generator and a plurality of gamma ray detectors; providing a second rack on another side of the vehicle, the second rack including a neutron generator and a plurality of gamma ray detectors; providing a control system, remote from the first and second racks, coupled to the neutron generators and gamma ray detectors; using the control system, causing the neutron generators to generate neutrons; and performing gamma ray spectroscopy on spectra read by the gamma ray detectors to look for a signature indicative of presence of an explosive. Various apparatus and other methods are also provided.

  13. Battery control and fault detection method

    SciTech Connect

    Bishop, W.S.

    1984-07-11

    This is a method for control, fault detection, fault isolation, and state-of-health monitoring of batteries and battery arrays. The method consists of measuring all of the battery, well, or cell group voltages, using statistics to determine a mean voltage and a standard deviation voltage, then comparing all of the measured voltages to the mean voltage. If the measured voltage deviates from the mean voltage by an arbitrary amount (number of standard deviations) corrective action can be implemented or an alarm signal given. The measurements need to be made rapidly enough to eliminate battery or cell voltage changes due to state of charge or temperature changes and, in most cases, require a computerized data collection/reduction system. Absolute high and/or low voltage limits can be included to prevent catastrophic events. The concept can be expanded to include similar temperature, pressure and/or battery current measurements in an array.

  14. Nuclease stability of boron-modified nucleic acids: application to label-free mismatch detection.

    PubMed

    Reverte, Maëva; Vasseur, Jean-Jacques; Smietana, Michael

    2015-11-21

    5'-End boronic acid-modified oligonucleotides were evaluated against various nucleases at single and double stranded levels. The results show that these modifications induce a high resistance to degradation by calf-spleen and snake venom phosphodiesterases. More importantly, this eventually led to the development of a new label-free enzyme-assisted fluorescence-based method for single mismatch detection.

  15. Covalent immobilization of ascorbate oxidase onto polycarbonate strip for L-ascorbic acid detection.

    PubMed

    Kannoujia, Dileep Kumar; Kumar, Saroj; Nahar, Pradip

    2012-10-01

    Herein, a simple and rapid method is described for detection of L-ascorbic acid by ascorbate oxidase immobilized onto polycarbonate strip pre-activated by 1-fluoro-2-nitro-4-azidobenzene in photochemical reaction. Covalent attachment of ascorbate oxidase was confirmed by XPS studies. The immobilized-ascorbate oxidase shows higher pH, thermal and storage stability in comparison to free enzyme.

  16. Methods for detecting teratogenic agents in man.

    PubMed Central

    Shepard, T H; Miller, J R

    1976-01-01

    At a multidiscipline international meeting sponsored by L'Institut de la Vie held at Guadeloupe in January 1974, current methods for detecting teratogenic agents were outlined and discussed. Recommendations of the participants of the conference were: recognize the limitations of the present defenses against teratogenic agents; educate the public and medical profession about the known human teratogenic agents; select for animal teratogenicity screening among new and existing agents by emphasizing substances to which the entire population will be exposed, agents to which pregnant women are exposed, viruses which are found to persist in the human fetus, and agents which have become suspect from clinical experience; recognize that nearly all compounds have a fetotoxic dose but that this does not imply teratogenicity; encourage the development of new, quick in vitro testing methods for detecting teratogenic agents; monitor for sudden increases in the frequency of specific malformations in newborn infants and in aborted fetuses; assure that expert multidiscipline committees are available to evaluate the threat when suspected teratogens are reported; improve teratology information storage and retrieval systems by record linkage of clinical data, linkage between computer systems, and universal identifier system for chemical compounds and congenical malformations; foster the exchange of data, particularly those held by the pharmaceutical industry. PMID:1269501

  17. New method to detect caries via fluorescence

    NASA Astrophysics Data System (ADS)

    Eberhart, J.; Frentzen, M.; Thoms, M.

    2007-07-01

    Caries, a common and widespread infectious disease, has to be detected as early as possible. Based on the need for an easy and handy tool for preventing invasive treatment a new fluorescence camera system has been developed. Using this camera the so-called porphyrins, metabolic products of oral pathogenic bacteria can be visualized. Thereby fluorophores are excited at a wavelength of 405nm by the built-in GaN-LEDs. Healthy and diseased dental hard tissues fluoresce in the green and in the red spectral range, respectively, thus allowing differentiation by coulor. To prove the reliability of this fluorescence camera system, freshly extracted teeth were examined. Three different methods of analysis were verified and compared to give information about the lesions (sensitivity & selectivity): The extent of the fluorescence area, the integral of the red/green ratio of the lesion and the maximum red/green ratio in the area of interest. Histological sections of the teeth served as reference. In addition, the camera was compared to a tip probe sensor already available on the market. In total, our results show that regarding the three different algorithms of analysis, the maximum of the red/green ratio is a preferential method to evaluate carious lesions. Sound tissue, enamel caries and dentin caries can be clearly distinguished. The new fluorescence camera is a handy, efficient and fast device in order to detect lesions and seems to be superior to the tip probe sensor regarding the positioning. Further studies are required.

  18. Method and apparatus for vapor detection

    NASA Technical Reports Server (NTRS)

    Lerner, Melvin (Inventor); Hood, Lyal V. (Inventor); Rommel, Marjorie A. (Inventor); Pettitt, Bruce C. (Inventor); Erikson, Charles M. (Inventor)

    1980-01-01

    The method disclosed herein may be practiced by passing the vapors to be sampled along a path with halogen vapor, preferably chlorine vapor, heating the mixed vapors to halogenate those of the sampled vapors subject to halogenation, removing unreacted halogen vapor, and then sensing the vapors for organic halogenated compounds. The apparatus disclosed herein comprises means for flowing the vapors, both sample and halogen vapors, into a common path, means for heating the mixed vapors to effect the halogenation reaction, means for removing unreacted halogen vapor, and a sensing device for sensing halogenated compounds. By such a method and means, the vapors of low molecular weight hydrocarbons, ketones and alcohols, when present, such as methane, ethane, acetone, ethanol, and the like are converted, at least in part, to halogenated compounds, then the excess halogen removed or trapped, and the resultant vapors of the halogenated compounds sensed or detected. The system is highly sensitive. For example, acetone in a concentration of 30 parts per billion (volume) is readily detected.

  19. Mid-ultraviolet light-emitting diode detects dipicolinic acid.

    SciTech Connect

    Bogart, Katherine Huderle Andersen; Lee, Stephen Roger; Temkin, Henryk; Crawford, Mary Hagerott; Dasgupta, Purnendu K.; Li, Qingyang; Allerman, Andrew Alan; Fischer, Arthur Joseph

    2005-06-01

    Dipicolinic acid (DPA, 2,6-pyridinedicarboxylic acid) is a substance uniquely present in bacterial spores such as that from anthrax (B. anthracis). It is known that DPA can be detected by the long-lived fluorescence of its terbium chelate; the best limit of detection (LOD) reported thus far using a large benchtop gated fluorescence instrument using a pulsed Xe lamp is 2 nM. We use a novel AlGaN light-emitting diode (LED) fabricated on a sapphire substrate that has peak emission at 291 nm. Although the overlap of the emission band of this LED with the absorption band of Tb-DPA ({lambda}{sub max} doublet: 273, 279 nm) is not ideal, we demonstrate that a compact detector based on this LED and an off-the-shelf gated photodetection module can provide an LOD of 0.4 nM, thus providing a basis for convenient early warning detectors.

  20. Highly sensitive detection of dipicolinic acid with a water-dispersible terbium-metal organic framework.

    PubMed

    Bhardwaj, Neha; Bhardwaj, Sanjeev; Mehta, Jyotsana; Kim, Ki-Hyun; Deep, Akash

    2016-12-15

    The sensitive detection of dipicolinic acid (DPA) is strongly associated with the sensing of bacterial organisms in food and many types of environmental samples. To date, the demand for a sensitive detection method for bacterial toxicity has increased remarkably. Herein, we investigated the DPA detection potential of a water-dispersible terbium-metal organic framework (Tb-MOF) based on the fluorescence quenching mechanism. The Tb-MOF showed a highly sensitive ability to detect DPA at a limit of detection of 0.04nM (linear range of detection: 1nM to 5µM) and also offered enhanced selectivity from other commonly associated organic molecules. The present study provides a basis for the application of Tb-MOF for direct, convenient, highly sensitive, and specific detection of DPA in the actual samples.

  1. Label-free functional nucleic acid sensors for detecting target agents

    DOEpatents

    Lu, Yi; Xiang, Yu

    2015-01-13

    A general methodology to design label-free fluorescent functional nucleic acid sensors using a vacant site approach and an abasic site approach is described. In one example, a method for designing label-free fluorescent functional nucleic acid sensors (e.g., those that include a DNAzyme, aptamer or aptazyme) that have a tunable dynamic range through the introduction of an abasic site (e.g., dSpacer) or a vacant site into the functional nucleic acids. Also provided is a general method for designing label-free fluorescent aptamer sensors based on the regulation of malachite green (MG) fluorescence. A general method for designing label-free fluorescent catalytic and molecular beacons (CAMBs) is also provided. The methods demonstrated here can be used to design many other label-free fluorescent sensors to detect a wide range of analytes. Sensors and methods of using the disclosed sensors are also provided.

  2. Application of carbon and hydrogen stable isotope analyses to detect exogenous citric acid in Japanese apricot liqueur.

    PubMed

    Akamatsu, Fumikazu; Oe, Takaaki; Hashiguchi, Tomokazu; Hisatsune, Yuri; Kawao, Takafumi; Fujii, Tsutomu

    2017-08-01

    Japanese apricot liqueur manufacturers are required to control the quality and authenticity of their liqueur products. Citric acid made from corn is the main acidulant used in commercial liqueurs. In this study, we conducted spiking experiments and carbon and hydrogen stable isotope analyses to detect exogenous citric acid used as an acidulant in Japanese apricot liqueurs. Our results showed that the δ(13)C values detected exogenous citric acid originating from C4 plants but not from C3 plants. The δ(2)H values of citric acid decreased as the amount of citric acid added increased, whether the citric acid originated from C3 or C4 plants. Commercial liqueurs with declared added acidulant provided higher δ(13)C values and lower δ(2)H values than did authentic liqueurs and commercial liqueurs with no declared added acidulant. Carbon and hydrogen stable isotope analyses are suitable as routine methods for detecting exogenous citric acid in Japanese apricot liqueur.

  3. Development of a bacterial bioassay for atrazine and cyanuric acid detection

    PubMed Central

    Hua, Anna; Gueuné, Hervé; Cregut, Mickaël; Thouand, Gérald; Durand, Marie-José

    2015-01-01

    The s-triazine herbicides are compounds which can disseminate into soils and water. Due to their toxic effects on living organisms, their concentrations in drinking water are legislated by WHO recommendations. Here we have developed for the first time, to the best of our knowledge, an alternative method for physicochemical quantification using two bioluminescent bacterial biosensors: E. coli SM003 for cyanuric acid detection and E. coli SM004 for both atrazine and cyanuric acid detection. The concentration of cyanuric acid detection for E. coli SM003 ranges from 7.83 μM to 2.89 mM, and for E. coli SM004 ranges from 0.22 to 15 μM. Moreover, atrazine detection by E. coli SM004 ranges from 1.08 to 15 μM. According to WHO recommendations, the cyanuric acid detection range is sensitive enough to discriminate between polluted and drinking water. Nevertheless, the detection of atrazine by E. coli SM004 is only applicable for high concentrations of contaminants. PMID:25852669

  4. Rapid Methods for High-Throughput Detection of Sulfoxides▿

    PubMed Central

    Shainsky, Janna; Derry, Netta-Lee; Leichtmann-Bardoogo, Yael; Wood, Thomas K.; Fishman, Ayelet

    2009-01-01

    Enantiopure sulfoxides are prevalent in drugs and are useful chiral auxiliaries in organic synthesis. The biocatalytic enantioselective oxidation of prochiral sulfides is a direct and economical approach for the synthesis of optically pure sulfoxides. The selection of suitable biocatalysts requires rapid and reliable high-throughput screening methods. Here we present four different methods for detecting sulfoxides produced via whole-cell biocatalysis, three of which were exploited for high-throughput screening. Fluorescence detection based on the acid activation of omeprazole was utilized for high-throughput screening of mutant libraries of toluene monooxygenases, but no active variants have been discovered yet. The second method is based on the reduction of sulfoxides to sulfides, with the coupled release and measurement of iodine. The availability of solvent-resistant microtiter plates enabled us to modify the method to a high-throughput format. The third method, selective inhibition of horse liver alcohol dehydrogenase, was used to rapidly screen highly active and/or enantioselective variants at position V106 of toluene ortho-monooxygenase in a saturation mutagenesis library, using methyl-p-tolyl sulfide as the substrate. A success rate of 89% (i.e., 11% false positives) was obtained, and two new mutants were selected. The fourth method is based on the colorimetric detection of adrenochrome, a back-titration procedure which measures the concentration of the periodate-sensitive sulfide. Due to low sensitivity during whole-cell screening, this method was found to be useful only for determining the presence or absence of sulfoxide in the reaction. The methods described in the present work are simple and inexpensive and do not require special equipment. PMID:19465532

  5. Odour Detection Methods: Olfactometry and Chemical Sensors

    PubMed Central

    Brattoli, Magda; de Gennaro, Gianluigi; de Pinto, Valentina; Loiotile, Annamaria Demarinis; Lovascio, Sara; Penza, Michele

    2011-01-01

    The complexity of the odours issue arises from the sensory nature of smell. From the evolutionary point of view olfaction is one of the oldest senses, allowing for seeking food, recognizing danger or communication: human olfaction is a protective sense as it allows the detection of potential illnesses or infections by taking into account the odour pleasantness/unpleasantness. Odours are mixtures of light and small molecules that, coming in contact with various human sensory systems, also at very low concentrations in the inhaled air, are able to stimulate an anatomical response: the experienced perception is the odour. Odour assessment is a key point in some industrial production processes (i.e., food, beverages, etc.) and it is acquiring steady importance in unusual technological fields (i.e., indoor air quality); this issue mainly concerns the environmental impact of various industrial activities (i.e., tanneries, refineries, slaughterhouses, distilleries, civil and industrial wastewater treatment plants, landfills and composting plants) as sources of olfactory nuisances, the top air pollution complaint. Although the human olfactory system is still regarded as the most important and effective “analytical instrument” for odour evaluation, the demand for more objective analytical methods, along with the discovery of materials with chemo-electronic properties, has boosted the development of sensor-based machine olfaction potentially imitating the biological system. This review examines the state of the art of both human and instrumental sensing currently used for the detection of odours. The olfactometric techniques employing a panel of trained experts are discussed and the strong and weak points of odour assessment through human detection are highlighted. The main features and the working principles of modern electronic noses (E-Noses) are then described, focusing on their better performances for environmental analysis. Odour emission monitoring carried out

  6. Odour detection methods: olfactometry and chemical sensors.

    PubMed

    Brattoli, Magda; de Gennaro, Gianluigi; de Pinto, Valentina; Loiotile, Annamaria Demarinis; Lovascio, Sara; Penza, Michele

    2011-01-01

    The complexity of the odours issue arises from the sensory nature of smell. From the evolutionary point of view olfaction is one of the oldest senses, allowing for seeking food, recognizing danger or communication: human olfaction is a protective sense as it allows the detection of potential illnesses or infections by taking into account the odour pleasantness/unpleasantness. Odours are mixtures of light and small molecules that, coming in contact with various human sensory systems, also at very low concentrations in the inhaled air, are able to stimulate an anatomical response: the experienced perception is the odour. Odour assessment is a key point in some industrial production processes (i.e., food, beverages, etc.) and it is acquiring steady importance in unusual technological fields (i.e., indoor air quality); this issue mainly concerns the environmental impact of various industrial activities (i.e., tanneries, refineries, slaughterhouses, distilleries, civil and industrial wastewater treatment plants, landfills and composting plants) as sources of olfactory nuisances, the top air pollution complaint. Although the human olfactory system is still regarded as the most important and effective "analytical instrument" for odour evaluation, the demand for more objective analytical methods, along with the discovery of materials with chemo-electronic properties, has boosted the development of sensor-based machine olfaction potentially imitating the biological system. This review examines the state of the art of both human and instrumental sensing currently used for the detection of odours. The olfactometric techniques employing a panel of trained experts are discussed and the strong and weak points of odour assessment through human detection are highlighted. The main features and the working principles of modern electronic noses (E-Noses) are then described, focusing on their better performances for environmental analysis. Odour emission monitoring carried out through

  7. Nucleic acid detection and quantification in the developing world.

    PubMed

    Huggett, Jim; Green, Clare; Zumla, Alimuddin

    2009-04-01

    Techniques using nucleic acid amplification have not had the same amount of impact on research and clinical diagnosis in the developing world as that observed in the West. This is unsurprising when the costs and infrastructure required to perform nucleic acid amplification are considered. Despite this, nucleic acid amplification is being increasingly used in both research and diagnosis in countries such as Zambia and Tanzania. Scientific research in the developing world is made possible through the support and development of the necessary laboratory infrastructure and the establishment of special transport for the reagents and samples. This has enabled world-leading country-relevant research to be performed by local scientists on subjects ranging from rapid diagnosis of infectious diseases to measuring the RNA gene expression in an immune response. Concomitantly, the challenge presented by the need for tests that are more appropriate for a resource-poor setting has led to a number of newer methodologies for nucleic acid detection, which can be tailored to be performed in the field without the need for training in molecular biology. As nucleic acid amplification techniques become both simpler and cheaper, their impact is likely to play an increasingly crucial role in research and diagnosis in the developing world.

  8. BeadCons: detection of nucleic acid sequences by flow cytometry.

    PubMed

    Horejsh, Douglas; Martini, Federico; Capobianchi, Maria Rosaria

    2005-11-01

    Molecular beacons are single-stranded nucleic acid structures with a terminal fluorophore and a distal, terminal quencher. These molecules are typically used in real-time PCR assays, but have also been conjugated with solid matrices. This unit describes protocols related to molecular beacon-conjugated beads (BeadCons), whose specific hybridization with complementary target sequences can be resolved by cytometry. Assay sensitivity is achieved through the concentration of fluorescence signal on discrete particles. By using molecular beacons with different fluorophores and microspheres of different sizes, it is possible to construct a fluid array system with each bead corresponding to a specific target nucleic acid. Methods are presented for the design, construction, and use of BeadCons for the specific, multiplexed detection of unlabeled nucleic acids in solution. The use of bead-based detection methods will likely lead to the design of new multiplex molecular diagnostic tools.

  9. Method and system for turbomachinery surge detection

    DOEpatents

    Faymon, David K.; Mays, Darrell C.; Xiong, Yufei

    2004-11-23

    A method and system for surge detection within a gas turbine engine, comprises: measuring the compressor discharge pressure (CDP) of the gas turbine over a period of time; determining a time derivative (CDP.sub.D ) of the measured (CDP) correcting the CDP.sub.D for altitude, (CDP.sub.DCOR); estimating a short-term average of CDP.sub.DCOR.sup.2 ; estimating a short-term average of CDP.sub.DCOR ; and determining a short-term variance of corrected CDP rate of change (CDP.sub.roc) based upon the short-term average of CDP.sub.DCOR and the short-term average of CDP.sub.DCOR.sup.2. The method and system then compares the short-term variance of corrected CDP rate of change with a pre-determined threshold (CDP.sub.proc) and signals an output when CDP.sub.roc >CDP.sub.proc. The method and system provides a signal of a surge within the gas turbine engine when CDP.sub.roc remains>CDP.sub.proc for pre-determined period of time.

  10. Hazard Detection Methods for Lunar Landing

    NASA Technical Reports Server (NTRS)

    Brady, Tye; Zimpfer, Doug; Robertson, Edward; Epp, Chirold; Paschall, Stephen

    2009-01-01

    The methods and experiences from the Apollo Program are fundamental building blocks for the development of lunar landing strategies for the Constellation Program. Each of the six lunar landing Apollo missions landed under near ideal lighting conditions. The astronauts visually performed terrain relative navigation while looking out of windows, and were greatly aided by external communication and well lit scenes. As the LM approached the landing site, the astronauts performed visual hazard detection and avoidance, also under near-ideal lighting conditions. The astronauts were looking out of the windows trying to the best of their ability to avoid rocks, slopes, and craters and find a safe landing location. NASA has expressed a desire for global lunar access for both crewed and robotic sortie lunar exploration missions (Cook, 2007) (Dale, 2006). Early NASA architecture studies have identified the lunar poles as desirable locations for early lunar missions. These polar missions have less than ideal lighting conditions and will significantly affect the way a crewed vehicle plans to land at such locales. Consequently, a variety of hazard identification methods should be considered for use by the crew to ensure a high degree of safety. This paper discusses such identification methods applicable to the poorly lit polar lunar environment, better ensuring global access for the soon to be designed Lunar Lander Vehicle (LLV).

  11. SKM-SNP: SNP markers detection method.

    PubMed

    Liu, Yang; Li, Mark; Cheung, Yiu M; Sham, Pak C; Ng, Michael K

    2010-04-01

    SKM-SNP, SNP markers detection program, is proposed to identify a set of relevant SNPs for the association between a disease and multiple marker genotypes. We employ a subspace categorical clustering algorithm to compute a weight for each SNP in the group of patient samples and the group of normal samples, and use the weights to identify the subsets of relevant SNPs that categorize these two groups. The experiments on both Schizophrenia and Parkinson Disease data sets containing genome-wide SNPs are reported to demonstrate the program. Results indicate that our method can find some relevant SNPs that categorize the disease samples. The online SKM-SNP program is available at http://www.math.hkbu.edu.hk/~mng/SKM-SNP/SKM-SNP.html.

  12. Material degradation detection by magnetic method

    SciTech Connect

    Yamaguchi, A.; Maeda, N.; Sugibayashi, T.

    1995-08-01

    To be able to evaluate the life of nuclear power plant becomes inevitable as the plant operating period extends. So, magnetic methods using Barkhausen noise (BHN) and B-H curve were applied to detect the degradation by fatigue and thermal aging. Low alloy steel (SA 508 cl.2) was fatigued, and duplex stainless steel (SCS 14A) was aged at 400 C. For the degradation by thermal aging, BHN and B-H curve were measured and good correlations between magnetic properties and aging time were obtained. For fatigue, BHN was measured at predetermined loading cycles and, at each predetermined cycle, the effect of stress or strain condition in the measurement was evaluated. The results showed that BHN was affected by the stress or strain condition in the measurement, the cause of which seemed to be the change of internal stress condition, and by identifying the measuring condition, good correlation between BHN and fatigue damage was obtained.

  13. Systems and methods for detecting neutrons

    DOEpatents

    Bross, Alan D.; Mellott, Kerry L.; Pla-Dalmau, Anna

    2005-08-09

    Systems and methods for detecting neutrons. One or more neutron-sensitive scintillators can be configured from a plurality of nano-sized particles, dopants and an extruded plastic material, such as polystyrene. The nano-sized particles can be compounded into the extruded plastic material with at least one dopant that permits the plastic material to scintillate. One or more plastic light collectors can be associated with a neutron-sensitive scintillator, such that the plastic light collector includes a central hole thereof. A wavelength-shifting fiber can then be located within the hole. The wavelength shifting (WLS) fiber absorbs scintillation light having a wavelength thereof and re-emits the light at a longer wavelength.

  14. Liquid chromatography detection unit, system, and method

    DOEpatents

    Derenzo, Stephen E.; Moses, William W.

    2015-10-27

    An embodiment of a liquid chromatography detection unit includes a fluid channel and a radiation detector. The radiation detector is operable to image a distribution of a radiolabeled compound as the distribution travels along the fluid channel. An embodiment of a liquid chromatography system includes an injector, a separation column, and a radiation detector. The injector is operable to inject a sample that includes a radiolabeled compound into a solvent stream. The position sensitive radiation detector is operable to image a distribution of the radiolabeled compound as the distribution travels along a fluid channel. An embodiment of a method of liquid chromatography includes injecting a sample that comprises radiolabeled compounds into a solvent. The radiolabeled compounds are then separated. A position sensitive radiation detector is employed to image distributions of the radiolabeled compounds as the radiolabeled compounds travel along a fluid channel.

  15. Solar cell anomaly detection method and apparatus

    NASA Technical Reports Server (NTRS)

    Miller, Emmett L. (Inventor); Shumka, Alex (Inventor); Gauthier, Michael K. (Inventor)

    1981-01-01

    A method is provided for detecting cracks and other imperfections in a solar cell, which includes scanning a narrow light beam back and forth across the cell in a raster pattern, while monitoring the electrical output of the cell to find locations where the electrical output varies significantly. The electrical output can be monitored on a television type screen containing a raster pattern with each point on the screen corresponding to a point on the solar cell surface, and with the brightness of each point on the screen corresponding to the electrical output from the cell which was produced when the light beam was at the corresponding point on the cell. The technique can be utilized to scan a large array of interconnected solar cells, to determine which ones are defective.

  16. Total Acid Value Titration of Hydrotreated Biomass Fast Pyrolysis Oil: Determination of Carboxylic Acids and Phenolics with Multiple End-Point Detection

    SciTech Connect

    Christensen, E.; Alleman, T. L.; McCormick, R. L.

    2013-01-01

    Total acid value titration has long been used to estimate corrosive potential of petroleum crude oil and fuel oil products. The method commonly used for this measurement, ASTM D664, utilizes KOH in isopropanol as the titrant with potentiometric end point determination by pH sensing electrode and Ag/AgCl reference electrode with LiCl electrolyte. A natural application of the D664 method is titration of pyrolysis-derived bio-oil, which is a candidate for refinery upgrading to produce drop in fuels. Determining the total acid value of pyrolysis derived bio-oil has proven challenging and not necessarily amenable to the methodology employed for petroleum products due to the different nature of acids present. We presented an acid value titration for bio-oil products in our previous publication which also utilizes potentiometry using tetrabutylammonium hydroxide in place of KOH as the titrant and tetraethylammonium bromide in place of LiCl as the reference electrolyte to improve the detection of these types of acids. This method was shown to detect numerous end points in samples of bio-oil that were not detected by D664. These end points were attributed to carboxylic acids and phenolics based on the results of HPLC and GC-MS studies. Additional work has led to refinement of the method and it has been established that both carboxylic acids and phenolics can be determined accurately. Use of pH buffer calibration to determine half-neutralization potentials of acids in conjunction with the analysis of model compounds has allowed us to conclude that this titration method is suitable for the determination of total acid value of pyrolysis oil and can be used to differentiate and quantify weak acid species. The measurement of phenolics in bio-oil is subject to a relatively high limit of detection, which may limit the utility of titrimetric methodology for characterizing the acidic potential of pyrolysis oil and products.

  17. Detection of cyclopiazonic acid (CPA) in maize by immunoassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cyclopiazonic acid (a-CPA) is a tremorgenic mycotoxin that is commonly produced by certain of the Aspergilli, in particular A. flavus, which is more widely known for production of the aflatoxins. Despite the fact that a-CPA may co-occur with aflatoxins, immunoassay-based methods for monitoring for C...

  18. A PCR assay for detection of acetic acid-tolerant lactic acid bacteria in acidic food products.

    PubMed

    Nakano, Shigeru; Matsumura, Atsushi; Yamada, Toshihiro

    2004-03-01

    A PCR assay for the detection of acetic acid-tolerant lactic acid bacteria in the genera of Lactobacillus and Pediococcus was developed in this study. Primers targeting the bacterial 16S rRNA gene were newly designed and used in this PCR assay. To determine the specificity of the assay, 56 different bacterial strains (of 33 genera), 2 fungi, 3 animals, and 4 plants were tested. Results were positive for most tested bacterial members of 16S rRNA gene-based phylogenetic groups (classified in the Lactobacillus casei and Pediococcus group), including Lactobacillus fructivorans, Lactobacillus brevis, Lactobacillus buchneri, Lactobacillus plantarum, and Lactobacillus paracasei. For all other bacterial strains and eukaryote tested, results were negative. Bacterial DNA for PCR was prepared with a simple procedure with the use of Chelex 100 resin from culture after growth in deMan Rogosa Sharpe broth (pH 6.0). To test this PCR assay for the monitoring of the acetic acid-tolerant lactic acid bacteria, L. fructivorans was inoculated into several acidic food as an indicator. Before the PCR, the inoculation of 10 to 50 CFU of bacteria per g of food was followed by a 28-h enrichment culture step, and the PCR assay allowed the detection of bacterial cells. Including the enrichment culture step, the entire PCR detection process can be completed within 30 h.

  19. Acetic Acid Detection Threshold in Synthetic Wine Samples of a Portable Electronic Nose

    PubMed Central

    Macías, Miguel Macías; Manso, Antonio García; Orellana, Carlos Javier García; Velasco, Horacio Manuel González; Caballero, Ramón Gallardo; Chamizo, Juan Carlos Peguero

    2013-01-01

    Wine quality is related to its intrinsic visual, taste, or aroma characteristics and is reflected in the price paid for that wine. One of the most important wine faults is the excessive concentration of acetic acid which can cause a wine to take on vinegar aromas and reduce its varietal character. Thereby it is very important for the wine industry to have methods, like electronic noses, for real-time monitoring the excessive concentration of acetic acid in wines. However, aroma characterization of alcoholic beverages with sensor array electronic noses is a difficult challenge due to the masking effect of ethanol. In this work, in order to detect the presence of acetic acid in synthetic wine samples (aqueous ethanol solution at 10% v/v) we use a detection unit which consists of a commercial electronic nose and a HSS32 auto sampler, in combination with a neural network classifier (MLP). To find the characteristic vector representative of the sample that we want to classify, first we select the sensors, and the section of the sensors response curves, where the probability of detecting the presence of acetic acid will be higher, and then we apply Principal Component Analysis (PCA) such that each sensor response curve is represented by the coefficients of its first principal components. Results show that the PEN3 electronic nose is able to detect and discriminate wine samples doped with acetic acid in concentrations equal or greater than 2 g/L. PMID:23262483

  20. Acetic acid detection threshold in synthetic wine samples of a portable electronic nose.

    PubMed

    Macías, Miguel Macías; Manso, Antonio García; Orellana, Carlos Javier García; Velasco, Horacio Manuel González; Caballero, Ramón Gallardo; Chamizo, Juan Carlos Peguero

    2012-12-24

    Wine quality is related to its intrinsic visual, taste, or aroma characteristics and is reflected in the price paid for that wine. One of the most important wine faults is the excessive concentration of acetic acid which can cause a wine to take on vinegar aromas and reduce its varietal character. Thereby it is very important for the wine industry to have methods, like electronic noses, for real-time monitoring the excessive concentration of acetic acid in wines. However, aroma characterization of alcoholic beverages with sensor array electronic noses is a difficult challenge due to the masking effect of ethanol. In this work, in order to detect the presence of acetic acid in synthetic wine samples (aqueous ethanol solution at 10% v/v) we use a detection unit which consists of a commercial electronic nose and a HSS32 auto sampler, in combination with a neural network classifier (MLP). To find the characteristic vector representative of the sample that we want to classify, first we select the sensors, and the section of the sensors response curves, where the probability of detecting the presence of acetic acid will be higher, and then we apply Principal Component Analysis (PCA) such that each sensor response curve is represented by the coefficients of its first principal components. Results show that the PEN3 electronic nose is able to detect and discriminate wine samples doped with acetic acid in concentrations equal or greater than 2 g/L.

  1. Sensor And Method For Detecting A Superstrate

    NASA Technical Reports Server (NTRS)

    Arndt, G. Dickey (Inventor); Cari, James R. (Inventor); Ngo, Phong H. (Inventor); Fink, Patrick W. (Inventor); Siekierski, James D. (Inventor)

    2006-01-01

    Method and apparatus are provided for determining a superstrate on or near a sensor, e.g., for detecting the presence of an ice superstrate on an airplane wing or a road. In one preferred embodiment, multiple measurement cells are disposed along a transmission line. While the present invention is operable with different types of transmission lines, construction details for a presently preferred coplanar waveguide and a microstrip waveguide are disclosed. A computer simulation is provided as part of the invention for predicting results of a simulated superstrate detector system. The measurement cells may be physically partitioned, nonphysically partitioned with software or firmware, or include a combination of different types of partitions. In one embodiment, a plurality of transmission lines are utilized wherein each transmission line includes a plurality of measurement cells. The plurality of transmission lines may be multiplexed with the signal from each transmission line being applied to the same phase detector. In one embodiment, an inverse problem method is applied to determine the superstrate dielectric for a transmission line with multiple measurement cells.

  2. A paper based microfluidic device for easy detection of uric acid using positively charged gold nanoparticles.

    PubMed

    Kumar, Anand; Hens, Abhiram; Arun, Ravi Kumar; Chatterjee, Monosree; Mahato, Kuldeep; Layek, Keya; Chanda, Nripen

    2015-03-21

    A paper based microfluidic device is fabricated that can rapidly detect very low concentrations of uric acid (UA) using 3,5,3',5'-tetramethyl benzidine (TMB), H2O2 and positively charged gold nanoparticles ((+)AuNPs). In the presence of (+)AuNPs, H2O2 reacts with TMB to produce a bluish-green colour which becomes colourless on reaction with UA. This colorimetric method can detect as low as 8.1 ppm of UA within <20 minutes on white filter paper. This technique provides an alternative way for UA detection.

  3. Comparison of conventional staining methods and monoclonal antibody-based methods for Cryptosporidium oocyst detection.

    PubMed Central

    Arrowood, M J; Sterling, C R

    1989-01-01

    The sensitivity and specificity of seven microscopy-based Cryptosporidium oocyst detection methods were compared after application to unconcentrated fecal smears. The seven methods were as follows: (i) a commercial acid-fast (AF) stain (VOLU-SOL) method, (ii) Truant auramine-rhodamine (AR) stain method, (iii) fluorescein-conjugated C1B3 monoclonal antibody (MAb) direct fluorescence method, (iv) OW3 MAb indirect fluorescence method, (v) biotinylated OW3 indirect fluorescence method, (vi) biotinylated OW3-indirect diaminobenzidine (DAB) method, and (vii) biotinylated OW3-aminoethylcarbazole (AEC) method. A total of 281 randomly collected Formalin-fixed fecal samples (submitted to the Maricopa County Health Department, Phoenix, Ariz.) and 30 known positives (Formalin-fixed and K2Cr2O7-preserved stools from our laboratory) were examined in a blind test; 32 of 311 samples (10.3%) were confirmed positive. Of the confirmed positives, 40.6% were identified by the AF method, 93.8% were identified by the AR method, 93.8% were identified by the C1B3 method, 81.3% were identified by the OW3-DAB method, 71.9% were identified by the OW3-AEC method, 100% were identified by the OW3 indirect fluorescence method, and 100% were identified by the biotinylated OW3 indirect fluorescence method. False-positives were encountered by the AF and AR methods (52.0 and 85.7% specificity, respectively), while no false-positives encountered by the MAb-based methods. Oocysts in infected tissue sections were easily detected by the MAb-based methods. Images PMID:2475523

  4. Amplified electrochemical detection of nucleic acid hybridization via selective preconcentration of unmodified gold nanoparticles.

    PubMed

    Li, Yuan; Tian, Rui; Zheng, Xingwang; Huang, Rongfu

    2016-08-31

    The common drawback of optical methods for rapid detection of nucleic acid by exploiting the differential affinity of single-/double-stranded nucleic acids for unmodified gold nanoparticles (AuNPs) is its relatively low sensitivity. In this article, on the basis of selective preconcentration of AuNPs unprotected by single-stranded DNA (ssDNA) binding, a novel electrochemical strategy for nucleic acid sequence identification assay has been developed. Through detecting the redox signal mediated by AuNPs on 1, 6-hexanedithiol blocked gold electrode, the proposed method is able to ensure substantial signal amplification and a low background current. This strategy is demonstrated for quantitative analysis of the target microRNA (let-7a) in human breast adenocarcinoma cells, and a detection limit of 16 fM is readily achieved with desirable specificity and sensitivity. These results indicate that the selective preconcentration of AuNPs for electrochemical signal readout can offer a promising platform for the detection of specific nucleic acid sequence.

  5. Excellent storage stability and sensitive detection of neurotoxin quinolinic acid.

    PubMed

    Singh, Ranjana; Kashyap, Sunayana; Kumar, Suveen; Abraham, Shiju; Gupta, Tejendra K; Kayastha, Arvind M; Malhotra, Bansi D; Saxena, Preeti Suman; Srivastava, Anchal; Singh, Ranjan K

    2017-04-15

    Quinolinic acid (QA) is a metabolite of tryptophan degradation obtained through kynurenine pathway, produced naturally in the mammalian brain as well as in the human cerebrospinal fluid. The presence of QA ~10-40µM is a clear indicator of many neurological disorders as well as deficiency of vitamin B6 in human being. In the present work; rapid, sensitive and cost-effective bio-electrodes were prepared to detect the trace amount of endogenous neurotoxin (QA). Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) studies were carried out to measure the electrochemical response of the fabricated bio-electrodes as a function of QA concentrations. These devices were found to exhibit desirable sensitivity of ~7.86mAμM(-1)cm(-2) in wide concentration range (6.5μM-65mM). The lower detection limit of this device is as low as 6.5μM and it has excellent storage stability of ~30 days. The capability of the proposed electrochemical bio-sensor was also checked to detect QA in the real samples (human serum). These results reveal that the use of this electrochemical bio-sensor may provide a potential platform for the detection of QA in the real samples for the prior detection of many diseases.

  6. Detection of a New Interstellar Molecule: Thiocyanic Acid HSCN

    NASA Astrophysics Data System (ADS)

    Halfen, D. T.; Ziurys, L. M.; Brünken, S.; Gottlieb, C. A.; McCarthy, M. C.; Thaddeus, P.

    2009-09-01

    A new interstellar molecule, HSCN (thiocyanic acid), an energetic isomer of the well-known species HNCS, has been detected toward Sgr B2(N) with the Arizona Radio Observatory 12 m telescope. Eight rotational transitions in the Ka = 0 ladder were observed in the 2 mm and 3 mm bands. Five consecutive transitions in the 3 mm band are unblended, but three in the 2 mm band are partially masked by lines of other molecules. The peak intensity of all eight transitions are well described by a rotational temperature that is in very good agreement with that of many other molecules in this source. The line width and radial velocity of HSCN match closely with those of the ground state isomer HNCS (isothiocyanic acid), HNCO (isocyanic acid), and HOCN (cyanic acid); preliminary maps indicate that all four molecules are similarly distributed in Sgr B2. Although HSCN is calculated to lie over 3000 K higher in energy than HNCS, its column density of 1.3 × 1013 cm-2 in Sgr B2(N) is only three times lower than that of HNCS. The fractional abundances of HSCN and HNCS relative to H2 are 4.5 × 10-12 and 1.1 × 10-11. By analogy with the isomeric pair HCN and HNC, these two sulfur-bearing isomers are plausibly formed from a common cation precursor.

  7. Plasmonics-Based Detection of Virus Using Sialic Acid Functionalized Gold Nanoparticles.

    PubMed

    Lee, Changwon; Wang, Peng; Gaston, Marsha A; Weiss, Alison A; Zhang, Peng

    2017-01-01

    Biosensor for the detection of virus was developed by utilizing plasmonic peak shift phenomenon of the gold nanoparticles and viral infection mechanism of hemagglutinin on virus and sialic acid on animal cells. The plasmonic peak of the colloidal gold nanoparticles changes with the aggregation of the particles due to the plasmonic interaction between nearby particles and the color of the colloidal nanoparticle solution changes from wine red to purple. Sialic acid reduced and stabilized colloidal gold nanoparticle aggregation is induced by the addition of viral particles in the solution due to the hemagglutinin-sialic acid interaction. In this work, sialic acid reduced and stabilized gold nanoparticles (d = 20.1 ± 1.8 nm) were synthesized by a simple one-pot, green method without chemically modifying sialic acid. The gold nanoparticles showed target-specific aggregation with viral particles via hemagglutinin-sialic acid binding. A linear correlation was observed between the change in optical density and dilution of chemically inactivated influenza B virus species. The detection limit of the virus dilution (hemagglutinination assay titer, 512) was shown to be 0.156 vol% and the upper limit of the linearity can be extended with the use of more sialic acid-gold nanoparticles.

  8. Detection of saccharides by reactive desorption electrospray ionization (DESI) using modified phenylboronic acids

    NASA Astrophysics Data System (ADS)

    Zhang, Yun; Chen, Hao

    2010-01-01

    We have reported previously a method for the detection of sugars via in-situ derivatization with phenylboronic acid PhB(OH)2 using reactive desorption electrospray ionization (DESI, Chen et al., Chem. Commun. (2006) 597-599). The present study describes an improved method that employs modified phenylboronic acids including 3-nitrophenylboronic acid and N-methyl-4-pyridineboronic acid iodide. In contrast to using PhB(OH)2, enhanced sensitivity of using 3-nitrophenylboronic acid was observed due to the stabilization of the resulting boronate ester anion by the electron-withdrawing nitro group and the limit of detections (LODs) for glucose in water using 3-nitrophenylbornic acid and phenylboronic acid were determined to be 0.11 mM and 0.40 mM, respectively. In the case of N-methyl-4-pyridineboronic acid iodide, the corresponding LOD is 6.9 [mu]M and the higher sensitivity obtained is attributed to the efficient ionization of both the reactive DESI reagent and reaction product since the precursor acid with a quaternary ammonium group is pre-charged. In this case, additional important features are found: (i) unlike using phenylboronic acid or 3-nitrophenylbornic acid, the experiment, performed in the positive ion mode, is applicable to neutral and acidic saccharide solutions, facilitating the analysis of biological fluids without the need to adjust pH; (ii) simply by changing the spray solvent from water to acetonitrile, the method can be used for direct glucose analyses of both urine and serum samples via online desalting, due to the low solubility of salts of these biofluids in the sprayed organic solvent; (iii) in comparison with other sugar derivatizing reagents such as the Girard's reagent T, the N-methyl-4-pyridineboronic acid iodide shows higher reactivity in the reactive DESI; and (iv) the ions of saccharide DESI reaction products undergo extensive ring or glycosidic bond cleavage upon CID, a feature that might be useful in the structure elucidation of

  9. Detection of the Rotational Spectrum of Sulfoxylic Acid (hosoh)

    NASA Astrophysics Data System (ADS)

    Crabtree, Kyle N.; Martinez, Oscar, Jr.; Barreau, Lou; McCarthy, Michael C.; Thorwirth, Sven

    2013-06-01

    Sulfoxylic acid (HOSOH) is a chemical intermediate that falls roughly midway in the oxidation states of sulfur, between its highly reduced (H_2S) and oxidixed (H_2SO_4) forms. It is likely formed during atmospheric oxidation of anthropogenic and natural sulfur emissions, and might also be produced by UV processing of circumstellar ices. Despite considerable theoretical work, no gas-phase spectra of sulfoxylic acid or any of its structural isomers have previously been observed. We report the detection of the rotational spectra of the C_2 and C_s rotamers of HOSOH using a combination of Fourier transform microwave spectroscopy and microwave-microwave double resonance techniques, guided by new high-level quantum chemical calculations of their structures. The present work enables radioastronomical searches for these species, and also lays the groundwork for further chemical studies of its gas-phase formation and spectroscopic studies of other H_2SO_2 isomers.

  10. Methods of staining target chromosomal DNA employing high complexity nucleic acid probes

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel; Kallioniemi, Ol'li-Pekka; Kallioniemi, Anne; Sakamoto, Masaru

    2006-10-03

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML), retinoblastoma, ovarian and uterine cancers, and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar but genetically different diseases, and for many prognostic and diagnostic applications.

  11. Radio frequency detection assembly and method for detecting radio frequencies

    SciTech Connect

    Cown, Steven H.; Derr, Kurt Warren

    2010-03-16

    A radio frequency detection assembly is described and which includes a radio frequency detector which detects a radio frequency emission produced by a radio frequency emitter from a given location which is remote relative to the radio frequency detector; a location assembly electrically coupled with the radio frequency detector and which is operable to estimate the location of the radio frequency emitter from the radio frequency emission which has been received; and a radio frequency transmitter electrically coupled with the radio frequency detector and the location assembly, and which transmits a radio frequency signal which reports the presence of the radio frequency emitter.

  12. Barriers on Breast Cancer Early Detection Methods

    PubMed Central

    Aksoy, Yasemin Erkal; Turfan, Esin Çeber; Sert, Ebru; Mermer, Gülengül

    2015-01-01

    . Conclusion Barriers against implementation of breast cancer screening methods in women were related to level of education and lack of adequate information about breast cancer screening, and symptoms of breast cancer. Women’s lack of information about signs, symptoms and treatment in the early stages of breast cancer needs to be eliminated. Health care providers may have a key role in increasing breast cancer early detection rates.

  13. Continuous-flow free acid monitoring method and system

    DOEpatents

    Strain, James E.; Ross, Harley H.

    1981-01-01

    A free acid monitoring method and apparatus is provided for continuously measuring the excess acid present in a process stream. The disclosed monitoring system and method is based on the relationship of the partial pressure ratio of water and acid in equilibrium with an acid solution at constant temperature. A portion of the process stream is pumped into and flows through the monitor under the influence of gravity and back to the process stream. A continuous flowing sample is vaporized at a constant temperature and the vapor is subsequently condensed. Conductivity measurements of the condensate produces a nonlinear response function from which the free acid molarity of the sample process stream is determined.

  14. Continuous-flow free acid monitoring method and system

    DOEpatents

    Strain, J.E.; Ross, H.H.

    1980-01-11

    A free acid monitoring method and apparatus is provided for continuously measuring the excess acid present in a process stream. The disclosed monitoring system and method is based on the relationship of the partial pressure ratio of water and acid in equilibrium with an acid solution at constant temperature. A portion of the process stream is pumped into and flows through the monitor under the influence of gravity and back to the process stream. A continuous flowing sample is vaporized at a constant temperature and the vapor is subsequently condensed. Conductivity measurements of the condensate produces a nonlinear response function from which the free acid molarity of the sample process stream is determined.

  15. Methods and kits for nucleic acid analysis using fluorescence resonance energy transfer

    DOEpatents

    Kwok, Pui-Yan; Chen, Xiangning

    1999-01-01

    A method for detecting the presence of a target nucleotide or sequence of nucleotides in a nucleic acid is disclosed. The method is comprised of forming an oligonucleotide labeled with two fluorophores on the nucleic acid target site. The doubly labeled oligonucleotide is formed by addition of a singly labeled dideoxynucleoside triphosphate to a singly labeled polynucleotide or by ligation of two singly labeled polynucleotides. Detection of fluorescence resonance energy transfer upon denaturation indicates the presence of the target. Kits are also provided. The method is particularly applicable to genotyping.

  16. Sulfuric acid thermoelectrochemical system and method

    DOEpatents

    Ludwig, Frank A.

    1989-01-01

    A thermoelectrochemical system in which an electrical current is generated between a cathode immersed in a concentrated sulfuric acid solution and an anode immersed in an aqueous buffer solution of sodium bisulfate and sodium sulfate. Reactants consumed at the electrodes during the electrochemical reaction are thermochemically regenerated and recycled to the electrodes to provide continuous operation of the system.

  17. Classification and adulteration detection of vegetable oils based on fatty acid profiles.

    PubMed

    Zhang, Liangxiao; Li, Peiwu; Sun, Xiaoman; Wang, Xuefang; Xu, Baocheng; Wang, Xiupin; Ma, Fei; Zhang, Qi; Ding, Xiaoxia

    2014-08-27

    The detection of adulteration of high priced oils is a particular concern in food quality and safety. Therefore, it is necessary to develop authenticity detection method for protecting the health of customers. In this study, fatty acid profiles of five edible oils were established by gas chromatography coupled with mass spectrometry (GC/MS) in selected ion monitoring mode. Using mass spectral characteristics of selected ions and equivalent chain length (ECL), 28 fatty acids were identified and employed to classify five kinds of edible oils by using unsupervised (principal component analysis and hierarchical clustering analysis), supervised (random forests) multivariate statistical methods. The results indicated that fatty acid profiles of these edible oils could classify five kinds of edible vegetable oils into five groups and are therefore employed to authenticity assessment. Moreover, adulterated oils were simulated by Monte Carlo method to establish simultaneous adulteration detection model for five kinds of edible oils by random forests. As a result, this model could identify five kinds of edible oils and sensitively detect adulteration of edible oil with other vegetable oils about the level of 10%.

  18. Rapid detection of Salmonella in pet food: design and evaluation of integrated methods based on real-time PCR detection.

    PubMed

    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.

  19. Methods of refining and producing isomerized fatty acid esters and fatty acids from natural oil feedstocks

    DOEpatents

    Snead, Thomas E.; Cohen, Steven A.; Gildon, Demond L.; Beltran, Leslie V.; Kunz, Linda A.; Pals, Tessa M.; Quinn, Jordan R; Behrends, Jr., Raymond T.; Bernhardt, Randal J.

    2016-07-05

    Methods are provided for refining natural oil feedstocks and producing isomerized esters and acids. The methods comprise providing a C4-C18 unsaturated fatty ester or acid, and isomerizing the fatty acid ester or acid in the presence of heat or an isomerization catalyst to form an isomerized fatty ester or acid. In some embodiments, the methods comprise forming a dibasic ester or dibasic acid prior to the isomerizing step. In certain embodiments, the methods further comprise hydrolyzing the dibasic ester to form a dibasic acid. In certain embodiments, the olefin is formed by reacting the feedstock in the presence of a metathesis catalyst under conditions sufficient to form a metathesized product comprising olefins and esters, separating the olefins from the esters in the metathesized product, and transesterifying the esters in the presence of an alcohol to form a transesterified product having unsaturated esters.

  20. Surface-crack detection by microwave methods

    NASA Technical Reports Server (NTRS)

    Feinstein, L.; Hruby, R.

    1967-01-01

    Microwave surface-crack detection system examines metallic surfaces with a noncontacting probe. The change in the microwave signal reflected from the surface under investigation is an indication of the existence of surface flaws. This technique can detect flaws and scratches as small as 100 microinches.

  1. An acid-sensing ion channel that detects ischemic pain.

    PubMed

    Naves, L A; McCleskey, E W

    2005-11-01

    Ischemic pain occurs when there is insufficient blood flow for the metabolic needs of an organ. The pain of a heart attack is the prototypical example. Multiple compounds released from ischemic muscle likely contribute to this pain by acting on sensory neurons that innervate muscle. One such compound is lactic acid. Here, we show that ASIC3 (acid-sensing ion channel #3) has the appropriate expression pattern and physical properties to be the detector of this lactic acid. In rats, it is expressed only in sensory neurons and then only on a minority (approximately 40%) of these. Nevertheless, it is expressed at extremely high levels on virtually all dorsal root ganglion sensory neurons that innervate the heart. It is extraordinarily sensitive to protons (Hill slope 4, half-activating pH 6.7), allowing it to readily respond to the small changes in extracellular pH (from 7.4 to 7.0) that occur during muscle ischemia. Moreover, both extracellular lactate and extracellular ATP increase the sensitivity of ASIC3 to protons. This final property makes ASIC3 a "coincidence detector" of three molecules that appear during ischemia, thereby allowing it to better detect acidosis caused by ischemia than other forms of systemic acidosis such as hypercapnia.

  2. Boron containing amino acid compounds and methods for their use

    DOEpatents

    Glass, John D.; Coderre, Jeffrey A.

    2000-01-01

    The present invention provides new boron containing amino acid compounds and methods for making these compounds by contacting melphalan or another nitrogen mustard derivative and sodium borocaptate. The present invention also provides a method of treating a mammal having a tumor by administering to the mammal a therapeutically effective amount of the new boron containing amino acid compounds.

  3. Boron containing amino acid compounds and methods for their use

    SciTech Connect

    Glass, J.D.; Coderre, J.A.

    2000-01-25

    The present invention provides new boron containing amino acid compounds and methods for making these compounds by contacting melphalan or another nitrogen mustard derivative and sodium borocaptate. The present invention also provides a method of treating a mammal having a tumor by administering to the mammal a therapeutically effective amount of the new boron containing amino acid compounds.

  4. Mutant fatty acid desaturase and methods for directed mutagenesis

    DOEpatents

    Shanklin, John; Whittle, Edward J.

    2008-01-29

    The present invention relates to methods for producing fatty acid desaturase mutants having a substantially increased activity towards substrates with fewer than 18 carbon atom chains relative to an unmutagenized precursor desaturase having an 18 carbon chain length specificity, the sequences encoding the desaturases and to the desaturases that are produced by the methods. The present invention further relates to a method for altering a function of a protein, including a fatty acid desaturase, through directed mutagenesis involving identifying candidate amino acid residues, producing a library of mutants of the protein by simultaneously randomizing all amino acid candidates, and selecting for mutants which exhibit the desired alteration of function. Candidate amino acids are identified by a combination of methods. Enzymatic, binding, structural and other functions of proteins can be altered by the method.

  5. Determination of ascorbic acid and carotenoids in food commodities by liquid chromatography with mass spectrometry detection.

    PubMed

    Frenich, A Garrido; Torres, M E Hernández; Vega, A Belmonte; Vidal, J L Martínez; Bolaños, P Plaza

    2005-09-21

    Two methods, one to determine ascorbic acid and one to determine lycopene and beta-carotene, in vegetables and fruits by liquid chromatography coupled with mass spectrometry (LC-MS) have been established. The chromatographic separation of the studied compounds and their MS parameters were optimized to improve selectivity and sensitivity. In both methods, separation was carried out with two coupled columns, first a C(18) and then a dC(18), using as mobile phase 70% methanol (0.005% acetic acid) and 30% acetic acid 0.05% for ascorbic acid determination and a mixture of methanol, tetrahydrofuran, and acetonitrile (60:30:10 v/v/v) for carotenoid analysis in isocratic mode. The molecular ion was selected for the quantification in selective ion monitoring (SIM) mode. Ascorbic acid was detected with electrospray ionization probe (ESI) in negative mode, while chemical ionization atmospheric pressure (APCI) in positive mode was used for the target carotenoids. The methodology for ascorbic acid analysis is based on an extraction with polytron using methanol and a mixture of methaphosphoric acid and acetic acid. Extraction of the carotenoids was carried out with tetrahydrofuran/methanol (1:1) (v/v). The proposed methods were applied, after their corresponding validations, to the analysis of four varieties of tomatoes, tomato in tin enriched and dried tomato, and to the analysis of mango and kiwi fruits, to compare the content in these compounds. Moreover, the influence of the process of freezing and the effect that the manipulation/preservation has in the content of ascorbic acid in tomato have also been studied.

  6. Synthetic oligonucleotide antigens modified with locked nucleic acids detect disease specific antibodies

    NASA Astrophysics Data System (ADS)

    Samuelsen, Simone V.; Solov’Yov, Ilia A.; Balboni, Imelda M.; Mellins, Elizabeth; Nielsen, Christoffer Tandrup; Heegaard, Niels H. H.; Astakhova, Kira

    2016-10-01

    New techniques to detect and quantify antibodies to nucleic acids would provide a significant advance over current methods, which often lack specificity. We investigate the potential of novel antigens containing locked nucleic acids (LNAs) as targets for antibodies. Particularly, employing molecular dynamics we predict optimal nucleotide composition for targeting DNA-binding antibodies. As a proof of concept, we address a problem of detecting anti-DNA antibodies that are characteristic of systemic lupus erythematosus, a chronic autoimmune disease with multiple manifestations. We test the best oligonucleotide binders in surface plasmon resonance studies to analyze binding and kinetic aspects of interactions between antigens and target DNA. These DNA and LNA/DNA sequences showed improved binding in enzyme-linked immunosorbent assay using human samples of pediatric lupus patients. Our results suggest that the novel method is a promising tool to create antigens for research and point-of-care monitoring of anti-DNA antibodies.

  7. Synthetic oligonucleotide antigens modified with locked nucleic acids detect disease specific antibodies

    PubMed Central

    Samuelsen, Simone V.; Solov’yov, Ilia A.; Balboni, Imelda M.; Mellins, Elizabeth; Nielsen, Christoffer Tandrup; Heegaard, Niels H. H.; Astakhova, Kira

    2016-01-01

    New techniques to detect and quantify antibodies to nucleic acids would provide a significant advance over current methods, which often lack specificity. We investigate the potential of novel antigens containing locked nucleic acids (LNAs) as targets for antibodies. Particularly, employing molecular dynamics we predict optimal nucleotide composition for targeting DNA-binding antibodies. As a proof of concept, we address a problem of detecting anti-DNA antibodies that are characteristic of systemic lupus erythematosus, a chronic autoimmune disease with multiple manifestations. We test the best oligonucleotide binders in surface plasmon resonance studies to analyze binding and kinetic aspects of interactions between antigens and target DNA. These DNA and LNA/DNA sequences showed improved binding in enzyme-linked immunosorbent assay using human samples of pediatric lupus patients. Our results suggest that the novel method is a promising tool to create antigens for research and point-of-care monitoring of anti-DNA antibodies. PMID:27775006

  8. Highly sensitive detection of cancer cells with an electrochemical cytosensor based on boronic acid functional polythiophene.

    PubMed

    Dervisevic, Muamer; Senel, Mehmet; Sagir, Tugba; Isik, Sevim

    2017-04-15

    The detection of cancer cells through important molecular recognition target such as sialic acid is significant for the clinical diagnosis and treatment. There are many electrochemical cytosensors developed for cancer cells detection but most of them have complicated fabrication processes which results in poor reproducibility and reliability. In this study, a simple, low-cost, and highly sensitive electrochemical cytosensor was designed based on boronic acid-functionalized polythiophene. In cytosensors fabrication simple single-step procedure was used which includes coating pencil graphite electrode (PGE) by means of electro-polymerization of 3-Thienyl boronic acid and Thiophen. Electrochemical impedance spectroscopy and cyclic voltammetry were used as an analytical methods to optimize and measure analytical performances of PGE/P(TBA0.5Th0.5) based electrode. Cytosensor showed extremely good analytical performances in detection of cancer cells with linear rage of 1×10(1) to 1×10(6) cellsmL(-1) exhibiting low detection limit of 10 cellsmL(-1) and incubation time of 10min. Next to excellent analytical performances, it showed high selectivity towards AGS cancer cells when compared to HEK 293 normal cells and bone marrow mesenchymal stem cells (BM-hMSCs). This method is promising for future applications in early stage cancer diagnosis.

  9. Method of analysis at the U.S. Geological Survey California Water Science Center, Sacramento Laboratory - determination of haloacetic acid formation potential, method validation, and quality-control practices

    USGS Publications Warehouse

    Zazzi, Barbara C.; Crepeau, Kathryn L.; Fram, Miranda S.; Bergamaschi, Brian A.

    2005-01-01

    An analytical method for the determination of haloacetic acid formation potential of water samples has been developed by the U.S. Geological Survey California Water Science Center Sacramento Laboratory. The haloacetic acid formation potential is measured by dosing water samples with chlorine under specified conditions of pH, temperature, incubation time, darkness, and residual-free chlorine. The haloacetic acids formed are bromochloroacetic acid, bromodichloroacetic acid, dibromochloroacetic acid, dibromoacetic acid, dichloroacetic acid, monobromoacetic acid, monochloroacetic acid, tribromoacetic acid, and trichloroacetic acid. They are extracted, methylated, and then analyzed using a gas chromatograph equipped with an electron capture detector. Method validation experiments were performed to determine the method accuracy, precision, and detection limit for each of the compounds. Method detection limits for these nine haloacetic acids ranged from 0.11 to 0.45 microgram per liter. Quality-control practices include the use of blanks, quality-control samples, calibration verification standards, surrogate recovery, internal standard, matrix spikes, and duplicates.

  10. System and method for detecting cells or components thereof

    DOEpatents

    Porter, Marc D.; Lipert, Robert J.; Doyle, Robert T.; Grubisha, Desiree S.; Rahman, Salma

    2009-01-06

    A system and method for detecting a detectably labeled cell or component thereof in a sample comprising one or more cells or components thereof, at least one cell or component thereof of which is detectably labeled with at least two detectable labels. In one embodiment, the method comprises: (i) introducing the sample into one or more flow cells of a flow cytometer, (ii) irradiating the sample with one or more light sources that are absorbed by the at least two detectable labels, the absorption of which is to be detected, and (iii) detecting simultaneously the absorption of light by the at least two detectable labels on the detectably labeled cell or component thereof with an array of photomultiplier tubes, which are operably linked to two or more filters that selectively transmit detectable emissions from the at least two detectable labels.

  11. [Measurement of the transport activities of bile salt export pump using chemiluminescence detection method].

    PubMed

    Yamaguchi, Kana; Murai, Tsuyoshi; Yabuuchi, Hikaru; Hui, Shu-Ping; Kurosawa, Takao

    2010-05-01

    Monovalent bile acids, such as taurine- and glycine-conjugated bile acids, are excreted into bile by bile salt export pumps (BSEP, ABCB11). Human BSEP (hBSEP) is physiologically important because it was identified as the gene responsible for the genetic disease: progressive familial intrahepatic cholestasis type 2 (PFIC-2). The evaluation of the inhibitory effect of hBSEP transport activity provides significant information for predicting toxic potential in the early phase of drug development. The role and function of hBSEP have been investigated by the examination of the ATP-dependent transport of radioactive isotopically (RI)-labeled bile acid such as a tritium labeled taurocholic acid, in membrane vesicles obtained from hBSEP-expressing cells. The chemiluminescence detection method using 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD) had been developed for a simple analysis of bile acids in human biological fluids. This method is extremely sensitive and it may be applicable for the measurements of bile acid transport activities by hBSEP vesicles without using RI-labeled bile acid. The present paper deals with an application of the chemiluminescence detection method using 3alpha-HSD with enzyme cycling method to the measurement of ATP-dependent transport activities of taurocholic acid (T-CA) in membrane vesicles obtained from hBSEP-expressing Sf9 cells. Calibration curves for T-CA was linear over the range from 10 to 400 pmol/ml. The values of the kinetic parameters for hBSEP vesicles obtained by the chemiluminescence detection method were comparable with the values of that obtained by liquid chromatography-mass spectrometry method. This assay method was highly useful for the measurements of bile acid transport activities.

  12. [Capillary electrophoresis analysis for glyphosate, glufosinate and aminomethylphosphonic acid with laser-induced fluorescence detection].

    PubMed

    Cao, Liwei; Liang, Siliu; Tan, Xiaofang; Meng, Jianxin

    2012-12-01

    A sensitive analytical method was developed for the simultaneous determination of glyphosate, glufosinate and aminomethylphosphonic acid by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). 5-(4,6-Dichlorotriazinyl) amino fluorescein (DTAF) was successfully applied to label the herbicides. The optimal derivatization reaction was carried out in boric acid buffer of pH 9.5 at 30 degrees C for 40 min. The baseline separation of the three derivatives could be accomplished using 30 mmol/L boric acid, 15 mmol/L Brij-35 (pH 9.5) as the running buffer. The detection limits (S/N = 3) for the glyphosate, glufosinate and aminomethylphosphonic acid were 3.21, 6.14, 1.99 ng/kg, respectively. Finally, the method was successfully applied to the analysis of environmental samples, and the three compounds were measured without any interference from real samples. The recoveries of the compounds in these samples were 91.3% - 106.0%. The method has the advantages of easiness and sensitivity, and can meet the requirement of the determination of the herbicide and metabolite residues in the environmental samples.

  13. Presence of human immunodeficiency virus nucleic acids in wastewater and their detection by polymerase chain reaction.

    PubMed Central

    Ansari, S A; Farrah, S R; Chaudhry, G R

    1992-01-01

    The human immunodeficiency virus type 1 (HIV-1) released by infected individuals or present in human and hospital wastes can potentially cause contamination problems. The presence of HIV-1 was investigated in 16 environmental samples, including raw wastewater, sludge, final effluent, soil, and pond water, collected from different locations. A method was developed to extract total nucleic acids in intact form directly from the raw samples or from the viral concentrates of the raw samples. The isolated nucleic acids were analyzed for the presence of HIV-1 by using in vitro amplification of the target sequences by the polymerase chain reaction (PCR) method. HIV-1-specific proviral DNA and viral RNA were detected in the extracted nucleic acids obtained from three wastewater samples by this method. The specificity of the PCR-amplified products was determined by Southern blot hybridization with an HIV-1-specific oligonucleotide probe, SK19. The isolated nucleic acids from wastewater samples were also screened for the presence of poliovirus type 1, representing a commonly found enteric virus, and simian immunodeficiency virus, representing, presumably, rare viruses. While poliovirus type 1 viral RNA was found in all of the wastewater samples, none of the samples yielded a simian immunodeficiency virus-specific product. No PCR-amplified product was yielded when wastewater samples were directly used for the detection of HIV-1 and poliovirus type 1. The wastewater constituents appeared to be inhibitory to the enzymes reverse transcriptase and DNA polymerase.(ABSTRACT TRUNCATED AT 250 WORDS) Images PMID:1476440

  14. Nuclear material detection apparatus and method

    DOEpatents

    Jones, James L.; Hoggan, Jerry M.; Harker, Yale D.; Yoon, Woo Y.; Johnson, Larry O.

    2006-11-28

    A device for detecting photonuclear-induced neutrons is described herein. One embodiment of the device may comprise a neutron detector and a detection circuit. The neutron detector may comprise a detector output. The detection circuit may be operatively connected to the detector output and may comprise an amplifier, a low-pass filter, and a high pass filter. The amplifier may comprise an amplifier input and an amplifier output. The amplifier input may be being operatively connected to the detector output. The low-pass filter may comprise a low-pass filter input and a low-pass filter output. The low-pass filter input may be operatively connected to the amplifier output. The high-pass filter may comprise a high-pass filter input and a high-pass filter output. The high-pass filter input may be operatively connected to the amplifier output.

  15. Source detection with the scaling index method.

    NASA Astrophysics Data System (ADS)

    Wiedemann, G.; Scheingraber, H.; Voges, W.

    The purpose of source detection algorithms in general is to distinguish between sources and non-significant background fluctuations. Most algorithms introduce an artificial partition of the event space by use of sliding windows. The procedure proposed here uses a natural partition given by the events themselves. The scaling index formalism assigns a scalar parameter to every event which can be classified by means of distribution theory as belonging to the background or to a source. No assumptions about the source geometry are necessary making the formalism most sensitive to the detection of extended irregular sources.

  16. Visual, base-specific detection of nucleic acid hybridization using polymerization-based amplification.

    PubMed

    Hansen, Ryan R; Johnson, Leah M; Bowman, Christopher N

    2009-03-15

    Polymerization-based signal amplification offers sensitive visualization of biotinylated biomolecules functionalized to glass microarrays in a manner suitable for point-of-care use. Here we report using this method for visual detection of multiplexed nucleic acid hybridizations from complex media and develop an application toward point mutation detection and single nucleotide polymorphism (SNP) typing. Primer extension reactions were employed to label selectively and universally all complementary surface DNA hybrids with photoinitiators, permitting simultaneous and dynamic photopolymerization from positive sites to 0.5-nM target concentrations. Dramatic improvements in signal ratios between complementary and mismatched hybrids enabled visual discrimination of single base differences in KRAS codon-12 biomarkers.

  17. Sequential injection titration with spectrophotometric detection for the assay of acidity in fruit juices.

    PubMed

    Jakmunee, Jaroon; Rujiralai, Thitima; Grudpan, Kate

    2006-01-01

    A simple sequential injection analysis (SIA) with spectrophotometric detection for an assay of acidity in fruit juice was investigated. An alkaline reagent (sodium hydroxide), a sample and an indicator (phenolphthalein) were first aspirated and stacked as adjacent zones in a holding coil. With flow reversal through a reaction coil to the detector, zone penetration occurred, leading to a neutralization reaction that caused a decrease in the color intensity of the indicator being monitored for absorbance at 552 nm. The effects of various parameters were studied. Linear calibration graphs for acidities of 0.2 - 1.0 and 0.5 - 2.5% w/v citric acid as a standard, with a relative standard deviation of 1% (acidity of 0.3 - 0.6% w/v as citric acid, n=11) and a sample throughput of 30 samples h(-1), were achieved. The developed method was validated by a standard titrimetric method for assaying the acidity of fruit juice samples.

  18. Compositions and method for controlling precipitation when acidizing sour wells

    SciTech Connect

    Dill, W.R.; Walker, M.L.

    1990-08-21

    This patent describes a method of treating a sour well penetrating a subterranean formation. It comprises: introducing into the well a treating fluid comprising an acid solution having a pH below 1.9, an iron sequestering agent comprising at least one compound selected from the group consisting of aminopolycarboxylic acids, hydroxycarboxylic acids, cyclic polyethers and derivatives of the acids and ethers, present in an amount of from about 0.25 to about 5 percent by weight of the acid solution, and a sulfide modifier comprising at least one compound selected from the group consisting of an aldehyde, acetal, hemiacetal and any other compound capable of forming aldehydes in the acid solution, present in an amount of from about 0.25 to about 5 percent of the acid solution; and treating the subterranean formation with the treating fluid.

  19. Determination of Cr(III) in chromic acid. Comparison of spectrophotometric and ion chromatographic methods

    SciTech Connect

    Smith, R.E.; Smith, C.H.

    1986-04-01

    Two methods have been developed for determining Cr(III) in chromic acid solutions. Both methods are based on the formation of a Cr-EDTA complex. The ion chromatographic method detects the Cr-EDTA as an anion using chemically suppressed conductivity. The spectrophotometric method detects the Cr-EDTA as a colored complex by measuring the absorbance at 540 nm. The conditions necessary for forming the Cr-EDTA complex are described. The results obtained by the spectrophotometric and ion chromatographic methods are compared. 15 refs., 5 figs., 1 tab.

  20. [Evaluation of phenotypic methods for the detection of KPC carbapenemases in Klebsiella pneumoniae].

    PubMed

    Nicola, Federico G; Nievas, Jimena; Smayevsky, Jorgelina

    2012-01-01

    We evaluated phenotypic methods for the detection of kPc carbapenemases in 44 clinical isolates of K. pneumoniae having reduced susceptibility to carbapenems, 30 of which were kPc-positive and 14 kPc-negative. Both the agar dilution and disk diffusion methods were performed for imipenem, meropenem and ertapenem. The following phenotypic methods were assayed: the double disk synergy test, using boronic acid or clavulanic acid as inhibitors, "combined" disks of carbapenem plus inhibitor (boronic acid, clavulanic acid and both boronic plus clavulanic acid), by using a pre-diffusion technique and the modified Hodge test. The double disk diffusion test using boronic acid could detect all kPc-positive isolates, but adjustment of disk distance was necessary for achieving such performance. The simulation of combined disks by our pre-diffusion technique detected all kPcpositive strains for all 3 carbapenems when using boronic acid as inhibitor, clavulanic acid was less susceptible and specific as compared with boronic acid. The modified Hodge test using any carbapenem was clearly positive for all kPc-producing isolates. This test was negative for all kPc-negative strains when imipenem or meropenem were used, but 2/14 isolates yielded a weak positive result when using ertapenem. By measuring the enhanced growth of E. coli aTcc 25922 observed in this test, we could objectively discriminate true-positive (= 8 mm) from false-positive results (< 5 mm). Our results show that the use of phenotypic methods is effective for the rapid detection of kPc producers in K. pneumoniae isolates with reduced susceptibility to carbapenems.

  1. A method for analysis of vanillic acid in polar ice cores

    NASA Astrophysics Data System (ADS)

    Grieman, M. M.; Greaves, J.; Saltzman, E. S.

    2015-02-01

    Biomass burning generates a wide range of organic compounds that are transported via aerosols to the polar ice sheets. Vanillic acid is a product of conifer lignin combustion, which has previously been observed in laboratory and ambient biomass burning aerosols. In this study a method was developed for analysis of vanillic acid in melted polar ice core samples. Vanillic acid was chromatographically separated using reversed-phase liquid chromatography (HPLC) and detected using electrospray ionization-triple quadrupole mass spectrometry (ESI-MS/MS). Using a 100 μL injection loop and analysis time of 4 min, we obtained a detection limit of 77 ppt (parts per trillion by mass) and an analytical precision of ±10%. Measurements of vanillic acid in Arctic ice core samples from the Siberian Akademii Nauk core are shown as an example application of the method.

  2. A reliable method for detecting complexed DNA in vitro

    NASA Astrophysics Data System (ADS)

    Holladay, C.; Keeney, M.; Newland, B.; Mathew, A.; Wang, W.; Pandit, A.

    2010-12-01

    Quantification of eluted nucleic acids is a critical parameter in characterizing biomaterial based gene-delivery systems. The most commonly used method is to assay samples with an intercalating fluorescent dye such as PicoGreen®. However, this technique was developed for unbound DNA and the current trend in gene delivery is to condense DNA with transfection reagents, which interfere with intercalation. Here, for the first time, the DNA was permanently labeled with the fluorescent dye Cy5 prior to complexation, an alternative technique hypothesized to allow quantification of both bound and unbound DNA. A comparison of the two methods was performed by quantifying the elution of six different varieties of DNA complexes from a model biomaterial (collagen) scaffold. After seven days of elution, the PicoGreen® assay only allowed detection of three types of complexes (those formed using Lipofectin™ and two synthesised copolymers). However, the Cy5 fluorescent labeling technique enabled detection of all six varieties including those formed via common transfection agents poly(ethylene imine), poly-l-lysine and SuperFect™. This allowed reliable quantification of the elution of all these complexes from the collagen scaffold. Thus, while intercalating dyes may be effective and reliable for detecting double-stranded, unbound DNA, the technique described in this work allowed reliable quantification of DNA independent of complexation state.Quantification of eluted nucleic acids is a critical parameter in characterizing biomaterial based gene-delivery systems. The most commonly used method is to assay samples with an intercalating fluorescent dye such as PicoGreen®. However, this technique was developed for unbound DNA and the current trend in gene delivery is to condense DNA with transfection reagents, which interfere with intercalation. Here, for the first time, the DNA was permanently labeled with the fluorescent dye Cy5 prior to complexation, an alternative technique

  3. Acidic Fluids Across Mars: Detections of Magnesium-Nickel Sulfates

    NASA Technical Reports Server (NTRS)

    Yen, A. S.; Ming, D. W.; Gellert, R.; Mittlefehldt, D. W.; Rampe, E. B.; Vaniman, D. T.; Thompson, L. M.; Morris, R. V.; Clark, B. C.; VanBommel, S. J.

    2017-01-01

    Calcium, magnesium and ferric iron sulfates have been detected by the instrument suites on the Mars rovers. A subset of the magnesium sulfates show clear associations with nickel. These associations indicate Ni(2+) co-precipitation with or substitution for Mg(2+) from sulfate-saturated solutions. Nickel is ex-tracted from primary rocks almost exclusively at pH values less than 6, constraining the formation of these Mg-Ni sulfates to mildly to strongly acidic conditions. There is clear evidence for aqueous alteration at the rim of Endeavour Crater (Meridiani Planum), in the Murray formation mudstone (Gale Crater), and near Home Plate (Gusev Crater). The discovery of Mg-Ni sulfates at these locations indicates a history of fluid-rock interactions at low pH.

  4. Lewis acid-assisted detection of nerve agents in water.

    PubMed

    Butala, Rahul R; Creasy, William R; Fry, Roderick A; McKee, Michael L; Atwood, David A

    2015-06-07

    The five-coordinate compound, Salen((t)Bu)Al(Ac), prepared in situ from Salen((t)Bu)AlBr and NH4Ac, forms Lewis acid-base adducts in aqueous solution with the G-type nerve agents, Sarin and Soman, and the VX hydrolysis product, ethylmethylphosphonate (EMPA). The resulting compounds, [Salen((t)Bu)Al(NA)](+)[Ac] (-) (with NA = Sarin, Soman, and EMPA) are sufficiently stable to be identified by ESI-MS. Molecular ion peaks were detected for every compound with little or no fragmentation. The distinctive MS signatures for the [Salen((t)Bu)Al(NA)](+) compounds provide a new technique for identifying nerve agents from aqueous solution. The energetics of the displacement of Ac(-) by the nerve agents to form [Salen((t)Bu)Al(NA)](+)[Ac](-) were determined computationally.

  5. Sensitive detection of nucleic acids with rolling circle amplification and surface-enhanced Raman scattering spectroscopy.

    PubMed

    Hu, Juan; Zhang, Chun-yang

    2010-11-01

    Detection of specific DNA sequences is important to molecular biology research and clinical diagnostics. To improve the sensitivity of surface-enhanced Raman scattering spectroscopy (SERS), a variety of signal amplification methods has been developed, including Raman-active-dye, polymerase chain reaction (PCR) technology, molecular beacon, SERS-active substrates, and SERS-tag. However, the combination of rolling circle amplification (RCA) with SERS for nucleic acid detection has not been reported. Herein, we describe a new approach for nucleic acid detection by the combination of RCA reaction with SERS. Because of the binding of abundance repeated sequences of RCA products with gold nanoparticle (Au NP) and Rox-modified detection probes, SERS signal is significantly amplified and the detection limit of 10.0 pM might be achieved. The sensitivity of RCA-based SERS has increased by as much as 3 orders of magnitude as compared to PCR-based SERS and is also comparable with or even exceeds that of both RCA-based electrochemical and RCA-based fluorescent methods. This RCA-based SERS might discriminate perfect matched target DNA from 1-base mismatched DNA with high selectivity. The high sensitivity and selectivity of RCA-based SERS makes it a potential tool for early diagnosis of gene-related disease and also offers a great promise for multiplexed assays with DNA microarrays.

  6. Fluorometric Method for Determination of Uric Acid in Flour

    DTIC Science & Technology

    1980-07-01

    of the excreta of adult Tribolium used in this study for natural infestation, accumulates in infested products providing a quantitative measure of past...by Adult Tribolium 8 4 FLUOROMETRIC METHOD FOR DETERMINATION OF URIC ACID IN FLOUR Introduction Chemical and microanalytical techniques have been...determining uric acid content.’ Approximately 18% of the total excreta from Tribolium confusum is uric acid.2 Various investigators have established

  7. Rapid Methods for the Detection of General Fecal Indicators

    EPA Science Inventory

    Specified that EPA should develop: appropriate and effective indicators for improving detection in a timely manner of pathogens in coastal waters appropriate, accurate, expeditious and cost-effective methods for the timely detection of pathogens in coastal waters

  8. A comparison of moving object detection methods for real-time moving object detection

    NASA Astrophysics Data System (ADS)

    Roshan, Aditya; Zhang, Yun

    2014-06-01

    Moving object detection has a wide variety of applications from traffic monitoring, site monitoring, automatic theft identification, face detection to military surveillance. Many methods have been developed across the globe for moving object detection, but it is very difficult to find one which can work globally in all situations and with different types of videos. The purpose of this paper is to evaluate existing moving object detection methods which can be implemented in software on a desktop or laptop, for real time object detection. There are several moving object detection methods noted in the literature, but few of them are suitable for real time moving object detection. Most of the methods which provide for real time movement are further limited by the number of objects and the scene complexity. This paper evaluates the four most commonly used moving object detection methods as background subtraction technique, Gaussian mixture model, wavelet based and optical flow based methods. The work is based on evaluation of these four moving object detection methods using two (2) different sets of cameras and two (2) different scenes. The moving object detection methods have been implemented using MatLab and results are compared based on completeness of detected objects, noise, light change sensitivity, processing time etc. After comparison, it is observed that optical flow based method took least processing time and successfully detected boundary of moving objects which also implies that it can be implemented for real-time moving object detection.

  9. Fast and Sensitive Method for Determination of Domoic Acid in Mussel Tissue

    PubMed Central

    Barbaro, Elena; Zangrando, Roberta; Barbante, Carlo; Gambaro, Andrea

    2016-01-01

    Domoic acid (DA), a neurotoxic amino acid produced by diatoms, is the main cause of amnesic shellfish poisoning (ASP). In this work, we propose a very simple and fast analytical method to determine DA in mussel tissue. The method consists of two consecutive extractions and requires no purification steps, due to a reduction of the extraction of the interfering species and the application of very sensitive and selective HILIC-MS/MS method. The procedural method was validated through the estimation of trueness, extract yield, precision, detection, and quantification limits of analytical method. The sample preparation was also evaluated through qualitative and quantitative evaluations of the matrix effect. These evaluations were conducted both on the DA-free matrix spiked with known DA concentration and on the reference certified material (RCM). We developed a very selective LC-MS/MS method with a very low value of method detection limit (9 ng g−1) without cleanup steps. PMID:26904720

  10. Real-Time Nucleic Acid Sequence-Based Amplification Assay for Detection of Hepatitis A Virus

    PubMed Central

    Abd El Galil, Khaled H.; El Sokkary, M. A.; Kheira, S. M.; Salazar, Andre M.; Yates, Marylynn V.; Chen, Wilfred; Mulchandani, Ashok

    2005-01-01

    A nucleic acid sequence-based amplification (NASBA) assay in combination with a molecular beacon was developed for the real-time detection and quantification of hepatitis A virus (HAV). A 202-bp, highly conserved 5′ noncoding region of HAV was targeted. The sensitivity of the real-time NASBA assay was tested with 10-fold dilutions of viral RNA, and a detection limit of 1 PFU was obtained. The specificity of the assay was demonstrated by testing with other environmental pathogens and indicator microorganisms, with only HAV positively identified. When combined with immunomagnetic separation, the NASBA assay successfully detected as few as 10 PFU from seeded lake water samples. Due to its isothermal nature, its speed, and its similar sensitivity compared to the real-time RT-PCR assay, this newly reported real-time NASBA method will have broad applications for the rapid detection of HAV in contaminated food or water. PMID:16269748

  11. Online SERS Detection of the 20 Proteinogenic L-Amino Acids Separated by Capillary Zone Electrophoresis

    PubMed Central

    Negri, Pierre; Schultz, Zachary D.

    2014-01-01

    A sheath-flow surface-enhanced Raman scattering (SERS) detector is demonstrated to provide chemical information enabling identification of the 20 proteinogenic L-amino acids separated by capillary zone electrophoresis (CZE). Amino acids were used to illustrate the chemical specificity of SERS detection from structurally related molecules. Analysis of the SERS electropherograms obtained from the separation and sequential online detection of six groups of structurally related amino acids shows that our sheath-flow SERS detector is able to resolve the characteristic Raman bands attributed to the amine, carboxyl, and side chain constituents. The results demonstrate the chemical information available from our detector and also provide insight into the nature of the analyte interaction with the silver SERS substrate. The spectra extracted from the SERS electropherogram of a mixture containing the 20 proteinogenic L-amino acids show unique signatures characteristic to each amino acid, thus enabling identification. The results presented here demonstrate the potential of this sheath-flow SERS detector as a general purpose method for high throughput characterization and identification following separations of complex biomolecular mixtures. PMID:25268706

  12. Separation and detection of amino acid metabolites of Escherichia coli in microbial fuel cell with CE.

    PubMed

    Wang, Wei; Ma, Lihong; Lin, Ping; Xu, Kaixuan

    2016-07-01

    In this work, CE-LIF was employed to investigate the amino acid metabolites produced by Escherichia coli (E. coli) in microbial fuel cell (MFC). Two peptides, l-carnosine and l-alanyl-glycine, together with six amino acids, cystine, alanine, lysine, methionine, tyrosine, arginine were separated and detected in advance by a CE-LIF system coupled with a homemade spontaneous injection device. The injection device was devised to alleviate the effect of electrical discrimination for analytes during sample injection. All analytes could be completely separated within 8 min with detection limits of 20-300 nmol/L. Then this method was applied to analyze the substrate solution containing amino acid metabolites produced by E. coli. l-carnosine, l-alanyl-glycine, and cystine were used as the carbon, nitrogen, and sulfur source for the E. coli culture in the MFC to investigate the amino acid metabolites during metabolism. Two MFCs were used to compare the activity of metabolism of the bacteria. In the sample collected at the running time 200 h of MFC, the amino acid methionine was discovered as the metabolite with the concentrations 23.3 μg/L.

  13. Compositions and methods for detecting Noonan syndrome

    DOEpatents

    Gelb, Bruce D [New York, NY; Tartaglia, Marco [Rome, IT; Pennacchio, Len [Walnut Creek, CA

    2012-07-17

    Diagnostic and therapeutic applications for Noonan Syndrome are described. The diagnostic and therapeutic applications are based on certain mutations in a RAS-specific guanine nucleotide exchange factor gene SOS1 or its expression product. The diagnostic and therapeutic applications are also based on certain mutations in a serine/threonine protein kinase gene RAF1 or its expression product thereof. Also described are nucleotide sequences, amino acid sequences, probes, and primers related to RAF1 or SOS1, and variants thereof, as well as host cells expressing such variants.

  14. Determination of Dornic Acidity as a Method to Select Donor Milk in a Milk Bank

    PubMed Central

    Garcia-Lara, Nadia Raquel; Escuder-Vieco, Diana; Chaves-Sánchez, Fernando; De la Cruz-Bertolo, Javier; Pallas-Alonso, Carmen Rosa

    2013-01-01

    Abstract Background Dornic acidity may be an indirect measurement of milk's bacteria content and its quality. There are no uniform criteria among different human milk banks on milk acceptance criteria. The main aim of this study is to report the correlation between Dornic acidity and bacterial growth in donor milk in order to validate the Dornic acidity value as an adequate method to select milk prior to its pasteurization. Materials and Methods From 105 pools, 4-mL samples of human milk were collected. Dornic acidity measurement and culture in blood and McConkey's agar cultures were performed. Based on Dornic acidity degrees, we classified milk into three quality categories: top quality (acidity <4°D), intermediate (acidity between 4°D and 7°D), and milk unsuitable to be consumed (acidity ≥8°D). Spearman's correlation coefficient was used to perform statistical analysis. Results Seventy percent of the samples had Dornic acidity under 4°D, and 88% had a value under 8°D. A weak positive correlation was observed between the bacterial growth in milk and Dornic acidity. The overall discrimination performance of Dornic acidity was higher for predicting growth of Gram-negative organisms. In milk with Dornic acidity of ≥4°D, such a measurement has a sensitivity of 100% for detecting all the samples with bacterial growth with Gram-negative bacteria of over 105 colony-forming units/mL. Conclusions The correlation between Dornic acidity and bacterial growth in donor milk is weak but positive. The measurement of Dornic acidity could be considered as a simple and economical method to select milk to pasteurize in a human milk bank based in quality and safety criteria. PMID:23373435

  15. Methods and systems for detection of radionuclides

    DOEpatents

    Coates, Jr., John T.; DeVol, Timothy A.

    2010-05-25

    Disclosed are materials and systems useful in determining the existence of radionuclides in an aqueous sample. The materials provide the dual function of both extraction and scintillation to the systems. The systems can be both portable and simple to use, and as such can beneficially be utilized to determine presence and optionally concentration of radionuclide contamination in an aqueous sample at any desired location and according to a relatively simple process without the necessity of complicated sample handling techniques. The disclosed systems include a one-step process, providing simultaneous extraction and detection capability, and a two-step process, providing a first extraction step that can be carried out in a remote field location, followed by a second detection step that can be carried out in a different location.

  16. Standardized Methods for Detection of Poliovirus Antibodies.

    PubMed

    Weldon, William C; Oberste, M Steven; Pallansch, Mark A

    2016-01-01

    Testing for neutralizing antibodies against polioviruses has been an established gold standard for assessing individual protection from disease, population immunity, vaccine efficacy studies, and other vaccine clinical trials. Detecting poliovirus specific IgM and IgA in sera and mucosal specimens has been proposed for evaluating the status of population mucosal immunity. More recently, there has been a renewed interest in using dried blood spot cards as a medium for sample collection to enhance surveillance of poliovirus immunity. Here, we describe the modified poliovirus microneutralization assay, poliovirus capture IgM and IgA ELISA assays, and dried blood spot polio serology procedures for the detection of antibodies against poliovirus serotypes 1, 2, and 3.

  17. Comparison of GC-MS and FTIR methods for quantifying naphthenic acids in water samples.

    PubMed

    Scott, Angela C; Young, Rozlyn F; Fedorak, Phillip M

    2008-11-01

    The extraction of bitumen from the oil sands in Canada releases toxic naphthenic acids into the process-affected waters. The development of an ideal analytical method for quantifying naphthenic acids (general formula C(n)H(2n+Z)O(2)) has been impeded by the complexity of these mixtures and the challenges of differentiating naphthenic acids from other naturally-occurring organic acids. The oil sands industry standard FTIR method was compared with a newly-developed GC-MS method. Naphthenic acids concentrations were measured in extracts of surface and ground waters from locations within the vicinity of and away from the oil sands deposits and in extracts of process-affected waters. In all but one case, FTIR measurements of naphthenic acids concentrations were greater than those determined by GC-MS. The detection limit of the GC-MS method was 0.01 mg L(-1) compared to 1 mg L(-1) for the FTIR method. The results indicated that the GC-MS method is more selective for naphthenic acids, and that the FTIR method overestimates their concentrations.

  18. Detection of Fatty Acids from Intact Microorganisms by Molecular Beam Static Secondary Ion Mass Spectrometry

    SciTech Connect

    Ingram, Jani Cheri; Lehman, Richard Michael; Bauer, William Francis; O'Connell, Sean Patrick; Colwell, Frederick Scott; Shaw, Andrew D.

    2003-06-01

    We report the use of a surface analysis approach, static secondary ion mass spectrometry (SIMS) equipped with a molecular (ReO4-) ion primary beam, to analyze the surface of intact microbial cells. SIMS spectra of 28 microorganisms were compared to fatty acid profiles determined by gas chromatographic analysis of transesterfied fatty acids extracted from the same organisms. The results indicate that surface bombardment using the molecular primary beam cleaved the ester linkage characteristic of bacteria at the glycerophosphate backbone of the phospholipid components of the cell membrane. This cleavage enables direct detection of the fatty acid conjugate base of intact microorganisms by static SIMS. The limit of detection for this approach is approximately 107 bacterial cells/cm2. Multivariate statistical methods were applied in a graded approach to the SIMS microbial data. The results showed that the full data set could initially be statistically grouped based upon major differences in biochemical composition of the cell wall. The gram-positive bacteria were further statistically analyzed, followed by final analysis of a specific bacterial genus that was successfully grouped by species. Additionally, the use of SIMS to detect microbes on mineral surfaces is demonstrated by an analysis of Shewanella oneidensis on crushed hematite. The results of this study provide evidence for the potential of static SIMS to rapidly detect bacterial species based on ion fragments originating from cell membrane lipids directly from sample surfaces.

  19. Method for incorporating radioactive phosphoric acid solutions in concrete

    DOEpatents

    Wolf, G.A.; Smith, J.W.; Ihle, N.C.

    1982-07-08

    A method for incorporating radioactive phosphoric acid solutions in concrete is described wherein the phosphoric acid is reacted with Ca(OH)/sub 2/ to form a precipitate of hydroxyapatite and the hydroxyapatite is mixed with Portland cement to form concrete.

  20. Method for incorporating radioactive phosphoric acid solutions in concrete

    DOEpatents

    Wolf, Gary A [Kennewick, WA; Smith, Jeffrey W [Lancaster, OH; Ihle, Nathan C [Walla Walla, WA

    1984-01-01

    A method for incorporating radioactive phosphoric acid solutions in concrete is described wherein the phosphoric acid is reacted with Ca(OH).sub.2 to form a precipitate of hydroxyapatite and the hydroxyapatite is mixed with portland cement to form concrete.

  1. STATISTICAL EVALUATION OF AN ANALYTICAL GC/MS METHOD FOR THE DETERMINATION OF LONG CHAIN FATTY ACIDS

    EPA Science Inventory

    In-depth evaluation of an analytical method to detect and quantify long chain fatty acids (C8 - C16) at trace level concentrations (25-1000 µg/l) is presented. The method requires derivatization of the acids with methanolic boron trifluoride, separation, and...

  2. Anion-exchange high-performance liquid chromatography with conductivity detection for the analysis of phytic acid in food.

    PubMed

    Talamond, P; Doulbeau, S; Rochette, I; Guyot, J P

    2000-02-25

    A sensitive method for the accurate determination of phytic acid in food samples is described. The proposed procedure involves the anion-exchange liquid chromatography with conductivity detection. Initially, two methods of determination of phytic acid were compared: absorptiometry and high-performance ion chromatography (HPIC) with chemically suppressed conductivity detector. Unlike most conventional methods involving precipitation by FeCl3, the simpler and more reliable HPIC assay avoids the numerous assumptions inherent in the iron precipitation and the accuracy is independent of the phytate content. The protocol was also applied to a survey of phytic acid concentration in some cereal, oil and legume seeds.

  3. Method for detection of extremely low concentration

    DOEpatents

    Andresen, Brian D.; Miller, Fred S.

    2002-01-01

    An ultratrace detector system for hand-held gas chromatography having high sensitivity, for example, to emissions generated during production of weapons, biological compounds, drugs, etc. The detector system is insensitive to water, air, helium, argon, oxygen, and CO.sub.2. The detector system is basically composed of a hand-held capillary gas chromatography (GC), an insulated heated redox-chamber, a detection chamber, and a vapor trap. For example, the detector system may use gas phase redox reactions and spectral absorption of mercury vapor. The gas chromatograph initially separates compounds that percolate through a bed of heated mercuric oxide (HgO) in a silica--or other metal--aerogel material which acts as an insulator. Compounds easily oxidized by HgO liberate atomic mercury that subsequently pass through a detection chamber which includes a detector cell, such as quartz, that is illuminated with a 254 nm ultra-violet (UV) mercury discharge lamp which generates the exact mercury absorption bands that are used to detect the liberated mercury atoms. Atomic mercury strongly absorbs 254 nm energy is therefore a specific signal for reducing compounds eluting from the capillary GC, whereafter the atomic mercury is trapped for example, in a silicon-aerogel trap.

  4. Using Amino Acid Physicochemical Distance Transformation for Fast Protein Remote Homology Detection

    PubMed Central

    Liu, Bin; Wang, Xiaolong; Chen, Qingcai; Dong, Qiwen; Lan, Xun

    2012-01-01

    Protein remote homology detection is one of the most important problems in bioinformatics. Discriminative methods such as support vector machines (SVM) have shown superior performance. However, the performance of SVM-based methods depends on the vector representations of the protein sequences. Prior works have demonstrated that sequence-order effects are relevant for discrimination, but little work has explored how to incorporate the sequence-order information along with the amino acid physicochemical properties into the prediction. In order to incorporate the sequence-order effects into the protein remote homology detection, the physicochemical distance transformation (PDT) method is proposed. Each protein sequence is converted into a series of numbers by using the physicochemical property scores in the amino acid index (AAIndex), and then the sequence is converted into a fixed length vector by PDT. The sequence-order information can be efficiently included into the feature vector with little computational cost by this approach. Finally, the feature vectors are input into a support vector machine classifier to detect the protein remote homologies. Our experiments on a well-known benchmark show the proposed method SVM-PDT achieves superior or comparable performance with current state-of-the-art methods and its computational cost is considerably superior to those of other methods. When the evolutionary information extracted from the frequency profiles is combined with the PDT method, the profile-based PDT approach can improve the performance by 3.4% and 11.4% in terms of ROC score and ROC50 score respectively. The local sequence-order information of the protein can be efficiently captured by the proposed PDT and the physicochemical properties extracted from the amino acid index are incorporated into the prediction. The physicochemical distance transformation provides a general framework, which would be a valuable tool for protein-level study. PMID:23029559

  5. Method of detecting and counting bacteria

    NASA Technical Reports Server (NTRS)

    Picciolo, G. L.; Chappelle, E. W. (Inventor)

    1976-01-01

    An improved method is provided for determining bacterial levels, especially in samples of aqueous physiological fluids. The method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of nonbacterial ATP. The bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay. A concentration technique is included to make the method more sensitive. It is particularly useful where the fluid to be measured contains an unknown or low bacteria count.

  6. Eco-friendly microextraction method for the quantitative speciation of 13 haloacetic acids in water.

    PubMed

    Cardador, María José; Gallego, Mercedes

    2014-05-02

    This paper describes the first micro liquid-liquid extraction (MLLE) gas chromatography-mass spectrometry (GC-MS) method for the speciation of emerging iodinated acetic acids, along with conventional chlorinated and brominated acids in water. The haloacetic acids (HAAs) were derivatised using 3 reagents for their methylation, both in aqueous and organic media. The acidic methanol derivatisation in aqueous medium provided the best efficiency, requiring minimal sample manipulation. The derivatisation yield was improved through the use of microwave energy that drastically reduced reaction time (2 min). The HAA methyl esters were finally extracted using 250 μL of methyl tert-butyl ether. This MLLE combined with the use of a large-volume sample injection coupled to a programmed temperature vaporiser-GC-MS improved the sensitivity of the method and minimised the generation of hazardous residues in accordance with the principles of "Green Chemistry". Detection and quantification limits (excepting tribromoacetic acid) within the range of 0.01-0.15 μg/L and 0.03-0.5 μg/L, respectively, were obtained and the relative standard deviation was lower than 10%. The eco-friendly method was applied to the speciation of the 13 HAAs in treated (chlorinated and chloraminated water) and untreated water. Up to 8 HAAs were found at detectable levels in treated water. The highly toxic monoiodoacetic acid was detected in almost all the chloraminated water.

  7. High-speed separation and detection of amino acids in laver using a short capillary electrophoresis system.

    PubMed

    Wang, Wei; Ma, Lihong; Yao, Fenzeng; Lin, Xiuli; Xu, Kaixuan

    2015-01-01

    A high-speed separation method of capillary MEKC with LIF detection had been developed for separation and determination of amino acids in laver. The CE system comprised a manual slotted-vial array (SVA) for sample introduction that could improve the separation efficiency by reducing injection volume. Using a capillary with 80 mm effective separation length, the separation conditions for amino acids were optimized. Applied with the separation electric field strength of 300 V/cm, the ten amino acids could be completely separated within 2.5 min with 10 mol/L Na2HPO4-NaOH buffer (pH = 11.5) including 30 mmol/L SDS. Theoretical plates for amino acids ranged from 72,000 to 40,000 (corresponding to 1.1-2.0 μm plate heights) and the detection limits were between 25 and 80 nmol/L. Finally, this method was applied to analyze the composition of amino acids in laver and eight known amino acids could be found in the sample. The contents of five amino acids, tyrosine, glutamic acid, glycine, lysine, and aspartic acid that could be completely separated in real sample were determined. The recoveries ranged from 82.3% to 123% that indicated the good reliability for this method in laver sample analysis.

  8. Method for early detection of infectious mononucleosis

    DOEpatents

    Willard, K.E.

    1982-08-10

    Early detection of infectious mononucleosis is carried out using a sample of human blood by isolating and identifying the presence of Inmono proteins in the sample from a two-dimensional protein map with the proteins being characterized by having isoelectric banding as measured in urea of about -16 to -17 with respect to certain isoelectric point standards and molecular mass of about 70 to 75 K daltons as measured in the presence of sodium dodecylsulfate containing polyacrylamide gels, the presence of the Inmono proteins being correlated with the existence of infectious mononucleosis.

  9. System and method for detecting gas

    DOEpatents

    Chow, Oscar Ken; Moulthrop, Lawrence Clinton; Dreier, Ken Wayne; Miller, Jacob Andrew

    2010-03-16

    A system to detect a presence of a specific gas in a mixture of gaseous byproducts comprising moisture vapor is disclosed. The system includes an electrochemical cell, a transport to deliver the mixture of gaseous byproducts from the electrochemical cell, a gas sensor in fluid communication with the transport, the sensor responsive to a presence of the specific gas to generate a signal corresponding to a concentration of the specific gas, and a membrane to prevent transmission of liquid moisture, the membrane disposed between the transport and the gas sensor.

  10. A novel method for isolating and analyzing organic acids in biological cultures

    SciTech Connect

    Polman, K.

    1995-12-31

    Fermentatively produced organic acids have significant potential as chemical feedstocks for the production of various commodity materials. Such acids include acetic and succinic acids. Fermentations frequently result in the simultaneous production of two or more organic acids, and often other fermentation products as well. This necessitates separation of these products from each other, so that quantification and purification can be achieved. A multitude of methodologies for the identification, purification, and quantitation of organic acids has been developed and described; both liquid and gas chromatography have been used for such separations. High-performance liquid chromatography (HPLC) media used for the separation of organic acids have included C18 columns, Aminex HPX-87H (ion-moderated partition resin), TEAP-Si 100 Polyol (strongly basic anion-exchange resin), Dowex 1 (cation-exchange column), Shodex Ionpak KC811, and others. Methodologies for HPLC analysis of organic acids also vary in these aspects: (1) Sample pretreatment (e.g., pretreatment with Sep-Pak C18 cartridges or with DEAE-Sephadex); (2) Mobile-phase composition (e.g., dilute sulfuric acid or formic acid); and (3) Method of organic acid detection (e.g., refractive index or light absorption). In this study, we present a methodology for isolating and quantifying organic acids found in fermentation broths. The methodology is simple, utilizes dual separation chemistries to effect an enhanced separation capacity, and is durable in terms of HPLC column life.

  11. Correlation studies on surface particle detection methods

    NASA Technical Reports Server (NTRS)

    Peterson, Ronald V.; White, James C.

    1988-01-01

    The accurate determination of dust levels on optical surfaces is necessary to assess sensor system performance. A comparison study was made on several particle measurement methods including those based on direct imaging and light scattering. The effectiveness of removing the particles from the surface prior to determining particle size distributions was also assessed. These studies revealed that some methods, especially those requiring particle removal before analysis, are subject to large systematic errors affecting particle size distributions. Thus, an understanding of the particle measurement methods employed is necessary before any surface cleanliness or obstruction value assignments are accepted as true representations of an optical surface contamination condition.

  12. [THE DETECTION OF CONTENT OF DIAGNOSTICALLY SIGNIFICANT FATTY ACIDS AND INDIVIDUAL TRIGLYCERIDES IN BIOLOGICAL MEDIUMS BASED ON INFRARED SPECTROMETRY].

    PubMed

    Kalinin, A V; Krasheninnikov, V N; Sviridov, A P; Titov, V N

    2015-11-01

    The content of clinically important fatty acids and individual triglycerides in food and biological mediums are traditionally detected by gas and fluid chromatography in various methodical modifications. The techniques are hard-to-get in laboratories of clinical biochemistry. The study was carried out to develop procedures and equipment for operative quantitative detection of concentration of fatty acids, primarily palmitic saturated fatty acid and oleic mono unsaturated fatty acid. Also detection was applied to sums ofpolyenoic (eicosapentaenoic and docosahexaenoic acid) fatty acids in biological mediums (cod-liver oil, tissues, blood plasma) using spectrometers of short-range infrared band of different types: with Fourier transform, diffraction and combined scattering. The evidences of reliable and reproducible quantitative detection offatty acids were received on the basis of technique of calibration (regression) by projection on latent structures using standard samples of mixtures of oils and fats. The evaluation is implemented concerning possibility of separate detection of content of palmitic and oleic triglycerides in mediums with presence of water The choice of technical conditions and mode of application of certain types of infrared spectrometers and techniques of their calibration is substantiated

  13. Human immunodeficiency virus trans-activator of transcription peptide detection via ribonucleic acid aptamer on aminated diamond biosensor

    NASA Astrophysics Data System (ADS)

    Rahim Ruslinda, A.; Wang, Xianfen; Ishii, Yoko; Ishiyama, Yuichiro; Tanabe, Kyosuke; Kawarada, Hiroshi

    2011-09-01

    The potential of ribonucleic acid (RNA) as both informational and ligand binding molecule have opened a scenario in the development of biosensors. An aminated diamond-based RNA aptasensor is presented for human immunodeficiency virus (HIV) trans-activator of transcription (Tat) peptide protein detection that not only gives a labeled or label-free detection method but also provides a reusable platform for a simple, sensitive, and selective detection of proteins. The immobilized procedure was based on the binding interaction between positively charged amine terminated diamond and the RNA aptamer probe molecules with the negatively charged surface carboxylic compound linker molecule such as terephthalic acid.

  14. Endotoxemia: methods of detection and clinical correlates.

    PubMed Central

    Hurley, J C

    1995-01-01

    As an assay for endotoxin, the Limulus amebocyte lysate assay has several desirable properties: sensitivity, specificity, and potential for adaptation to a quantitative format. Several modifications have been developed to enhance its potential for clinical application. The modifications that allow quantitative measurement of endotoxin and also improve its application to blood samples are described in this review. In fluids other than blood, the detection of endotoxin with the Limulus amebocyte lysate assay can be used as an aid to identify the presence of gram-negative bacteria, and the assay has established utility. With blood, however, there are a range of factors that interfere with the detection of endotoxemia and there are disparate views with respect to the diagnostic and prognostic significance of the test results. In general, the clinical significance of the finding of endotoxemia broadly parallels the frequency and importance of gram-negative sepsis in the patient groups studied and a decline in endotoxin levels accompanies clinical improvement. However, with therapies designed to reduce levels of endotoxin, or to antagonize its effects, it is unclear whether clinical improvement occurs as a consequence of changes in the levels of endotoxemia. PMID:7621402

  15. Polarization sensitive optical coherence tomography detection method

    SciTech Connect

    Everett, M J; Sathyam, U S; Colston, B W; DaSilva, L B; Fried, D; Ragadio, J N; Featherstone, J D B

    1999-05-12

    This study demonstrates the potential of polarization sensitive optical coherence tomography (PS-OCT) for non-invasive in vivo detection and characterization of early, incipient caries lesions. PS-OCT generates cross-sectional images of biological tissue while measuring the effect of the tissue on the polarization state of incident light. Clear discrimination between regions of normal and demineralized enamel is first shown in PS-OCT images of bovine enamel blocks containing well-characterized artificial lesions. High-resolution, cross-sectional images of extracted human teeth are then generated that clearly discriminate between the normal and carious regions on both the smooth and occlusal surfaces. Regions of the teeth that appeared to be demineralized in the PS-OCT images were verified using histological thin sections examined under polarized light microscopy. The PS-OCT system discriminates between normal and carious regions by measuring the polarization state of the back-scattered 1310 nm light, which is affected by the state of demineralization of the enamel. Demineralization of enamel increases the scattereing coefficient, thus depolarizing the incident light. This study shows that PS-OCT has great potential for the detection, characterization, and monitoring of incipient caries lesions.

  16. Methods and compositions for efficient nucleic acid sequencing

    DOEpatents

    Drmanac, Radoje

    2006-07-04

    Disclosed are novel methods and compositions for rapid and highly efficient nucleic acid sequencing based upon hybridization with two sets of small oligonucleotide probes of known sequences. Extremely large nucleic acid molecules, including chromosomes and non-amplified RNA, may be sequenced without prior cloning or subcloning steps. The methods of the invention also solve various current problems associated with sequencing technology such as, for example, high noise to signal ratios and difficult discrimination, attaching many nucleic acid fragments to a surface, preparing many, longer or more complex probes and labelling more species.

  17. Methods and compositions for efficient nucleic acid sequencing

    DOEpatents

    Drmanac, Radoje

    2002-01-01

    Disclosed are novel methods and compositions for rapid and highly efficient nucleic acid sequencing based upon hybridization with two sets of small oligonucleotide probes of known sequences. Extremely large nucleic acid molecules, including chromosomes and non-amplified RNA, may be sequenced without prior cloning or subcloning steps. The methods of the invention also solve various current problems associated with sequencing technology such as, for example, high noise to signal ratios and difficult discrimination, attaching many nucleic acid fragments to a surface, preparing many, longer or more complex probes and labelling more species.

  18. Detection of the in vivo conversion of 2-pyrrolidinone to gamma-aminobutyric acid in mouse brain.

    PubMed

    Callery, P S; Stogniew, M; Geelhaar, L A

    1979-01-01

    Labeled gamma-aminobutyric acid was detected in mouse brain following intravenous injections of deuterium labeled 2-pyrrolidinone. [2H6]Pyrrolidinone was prepared by the reduction of [2H4]succinimide with lithium aluminum deuteride. Quantification was accomplished by a gas chromatography mass spectrometry assay method. gamma-Aminobutyric acid and internal standard, 5-aminovaleric acid, were converted to volatile derivatives by treatment with N,N-dimethylformamide dimethyl acetal. Quantitative estimates were derived from peak area measurements obtained from monitoring the parent ions of the gamma-aminobutyric acid and internal standard derivatives by repetitive scanning during the GC run. The conversion of pyrrolidinone to gamma-aminobutyric acid may provide a method for labeling central gamma-aminobutyric acid pools.

  19. Fe-nitrilotriacetic acid coordination polymer nanowires: an effective sensing platform for fluorescence-enhanced nucleic acid detection

    NASA Astrophysics Data System (ADS)

    Zhou, Yunchun; Liu, Qian; Sun, Xuping; Kong, Rongmei

    2017-02-01

    The determination of specific nucleic acid sequences is key in identifying disease-causing pathogens and genetic diseases. In this paper we report the utilization of Fe-nitrilotriacetic acid coordination polymer nanowires as an effective nanoquencher for fluorescence-enhanced nucleic acid detection. The detection is fast and the whole process can be completed within 15 min. This nanosensor shows a low detection limit of 0.2 nM with selectivity down to single-base mismatch. This work provides us with an attractive sensing platform for applications.

  20. Detection methods and performance criteria for genetically modified organisms.

    PubMed

    Bertheau, Yves; Diolez, Annick; Kobilinsky, André; Magin, Kimberly

    2002-01-01

    Detection methods for genetically modified organisms (GMOs) are necessary for many applications, from seed purity assessment to compliance of food labeling in several countries. Numerous analytical methods are currently used or under development to support these needs. The currently used methods are bioassays and protein- and DNA-based detection protocols. To avoid discrepancy of results between such largely different methods and, for instance, the potential resulting legal actions, compatibility of the methods is urgently needed. Performance criteria of methods allow evaluation against a common standard. The more-common performance criteria for detection methods are precision, accuracy, sensitivity, and specificity, which together specifically address other terms used to describe the performance of a method, such as applicability, selectivity, calibration, trueness, precision, recovery, operating range, limit of quantitation, limit of detection, and ruggedness. Performance criteria should provide objective tools to accept or reject specific methods, to validate them, to ensure compatibility between validated methods, and be used on a routine basis to reject data outside an acceptable range of variability. When selecting a method of detection, it is also important to consider its applicability, its field of applications, and its limitations, by including factors such as its ability to detect the target analyte in a given matrix, the duration of the analyses, its cost effectiveness, and the necessary sample sizes for testing. Thus, the current GMO detection methods should be evaluated against a common set of performance criteria.

  1. Sciatica: Detection and Confirmation by New Method

    PubMed Central

    Nadkarni, Sunil

    2014-01-01

    We need to overcome limitations of present assessment and also integrate newer research in our work about sciatica. Inflammation induces changes in the DRG and nerve root. It sensitizes the axons. Nociceptor is a unique axon. It is pseudo unipolar: both its ends, central and peripheral, behave in similar fashion. The nerve in periphery which carries these axons may selectively become sensitive to mechanical pressure--“mechanosensitized,” as we coin the phrase. Many pain questionnaires are used and are effective in identifying neuropathic pain solely on basis of descriptors but they do not directly physically correlate nerve root and pain. A thorough neurological evaluation is always needed. Physical examination is not direct pain assessment but testing mobility of nerve root and its effect on pain generation. There is a dogmatic dominance of dermatomes in assessment of leg pain. They are unreliable. Images may not correlate with symptoms and pathology in about 28% of cases. Electrophysiology may be normal in purely inflamed nerve root. Palpation may help in such inflammatory setting to refine our assessment further. Confirmation of sciatica is done by selective nerve root block (SNRB) today but it is fraught with several complications and needs elaborate inpatient and operating room set up. We have used the unique property of the pseudo unipolar axon that both its ends have similar functional properties and so inject along its peripheral end sodium channel blockers to block the basic cause of the mechanosensitization namely upregulated sodium channels in the root or DRG. Thus using palpation we may be able to detect symptomatic nerve in stage of inflammation and with distal end injection, along same inflamed nerve we may be able to abolish and so confirm sciatica. Discussions of sciatica pain diagnosis tend to immediately shift and centre on the affected disc rather than the nerve. Theoretically it may be possible to detect the affected nerve by palpating the

  2. Label-free amino acid detection based on nanocomposites of graphene oxide hybridized with gold nanoparticles.

    PubMed

    Zhang, Qian; Zhang, Diming; Lu, Yanli; Xu, Gang; Yao, Yao; Li, Shuang; Liu, Qingjun

    2016-03-15

    Nanocomposites of graphene oxide and gold nanoparticles (GO/GNPs) were synthesized for label-free detections of amino acids. Interactions between the composites and amino acids were investigated by both naked-eye observation and optical absorption spectroscopy. The GO/GNPs composites displayed apparent color changes and absorption spectra changes in presences of amino acids including glutamate, aspartate, and cysteine. The interaction mechanisms of the composites and amino acids were discussed and explored with sulfhydryl groups and non-α-carboxylic groups on the amino acids. Sensing properties of the composites were tested, while pure gold particles were used as the control. The results suggested that the GO/GNPs composites had better linearity and stability in dose-dependent responses to the amino acids than those of the particles, especially in detections for acidic amino acids. Therefore, the nanocomposites platform can provide a convenient and efficient approach for label-free optical detections of important molecules such as amino acids.

  3. Methods for detection of ataxia telangiectasia mutations

    DOEpatents

    Gatti, Richard A.

    2005-10-04

    The present invention is directed to a method of screening large, complex, polyexonic eukaryotic genes such as the ATM gene for mutations and polymorphisms by an improved version of single strand conformation polymorphism (SSCP) electrophoresis that allows electrophoresis of two or three amplified segments in a single lane. The present invention also is directed to new mutations and polymorphisms in the ATM gene that are useful in performing more accurate screening of human DNA samples for mutations and in distinguishing mutations from polymorphisms, thereby improving the efficiency of automated screening methods.

  4. Validation of two methods for fatty acids analysis in eggs.

    PubMed

    Mazalli, Mônica R; Bragagnolo, Neura

    2007-05-01

    A comparative study between two methods (lipid extraction followed by saponification and methylation, and direct methylation) to determine the fatty acids in egg yolk was evaluated. Direct methylation of the samples resulted in lower fatty acid content and greater variation in the results than the lipid extraction followed by saponification and methylation. The low repeatability observed for the direct HCl methylation method was probably due to a less efficient extraction and conversion of the fatty acids into their methyl esters as compared to the same procedure starting with the lipid extract. As the lipid extraction followed by esterification method was shown to be more precise it was validated using powdered egg certified as reference material (RM 8415, NIST) and applied to samples of egg, egg enriched with polyunsaturated omega-3 fatty acids (n-3 PUFA), and commercial spray-dried whole egg powder.

  5. Fatty acids determination in Bronte pistachios by gas chromatographic method.

    PubMed

    Pantano, Licia; Lo Cascio, Giovanni; Alongi, Angelina; Cammilleri, Gaetano; Vella, Antonio; Macaluso, Andrea; Cicero, Nicola; Migliazzo, Aldo; Ferrantelli, Vincenzo

    2016-10-01

    A gas chromatographic with flame ionization detector (GC-MS FID) method for the identification and quantification of fatty acids based on the extraction of lipids and derivatisation of free acids to form methyl esters was developed and validated. The proposed method was evaluated to a number of standard FAs, and Bronte pistachios samples were used for that purpose and to demonstrate the applicability of the proposed method. In this regard, repeatability, mean and standard deviation of the analytical procedure were calculated. The results obtained have demonstrated oleic acid as the main component of Bronte pistachios (72.2%) followed by linoleic acid (13.4%) and showed some differences in composition with respect to Tunisian, Turkish and Iranian pistachios.

  6. A numerical method of detecting singularity

    NASA Technical Reports Server (NTRS)

    Laporte, M.; Vignes, J.

    1978-01-01

    A numerical method is reported which determines a value C for the degree of conditioning of a matrix. This value is C = 0 for a singular matrix and has progressively larger values for matrices which are increasingly well-conditioned. This value is C sub = C max sub max (C defined by the precision of the computer) when the matrix is perfectly well conditioned.

  7. Comparison of methods for acid quantification: impact of resist components on acid-generating efficiency

    NASA Astrophysics Data System (ADS)

    Cameron, James F.; Fradkin, Leslie; Moore, Kathryn; Pohlers, Gerd

    2000-06-01

    Chemically amplified deep UV (CA-DUV) positive resists are the enabling materials for manufacture of devices at and below 0.18 micrometer design rules in the semiconductor industry. CA-DUV resists are typically based on a combination of an acid labile polymer and a photoacid generator (PAG). Upon UV exposure, a catalytic amount of a strong Bronsted acid is released and is subsequently used in a post-exposure bake step to deprotect the acid labile polymer. Deprotection transforms the acid labile polymer into a base soluble polymer and ultimately enables positive tone image development in dilute aqueous base. As CA-DUV resist systems continue to mature and are used in increasingly demanding situations, it is critical to develop a fundamental understanding of how robust these materials are. One of the most important factors to quantify is how much acid is photogenerated in these systems at key exposure doses. For the purpose of quantifying photoacid generation several methods have been devised. These include spectrophotometric methods, ion conductivity methods and most recently an acid-base type titration similar to the standard addition method. This paper compares many of these techniques. First, comparisons between the most commonly used acid sensitive dye, tetrabromophenol blue sodium salt (TBPB) and a less common acid sensitive dye, Rhodamine B base (RB) are made in several resist systems. Second, the novel acid-base type titration based on the standard addition method is compared to the spectrophotometric titration method. During these studies, the make up of the resist system is probed as follows: the photoacid generator and resist additives are varied to understand the impact of each of these resist components on the acid generation process.

  8. An edge detection method for strong noisy image using shearlets

    NASA Astrophysics Data System (ADS)

    Li, Yuming; Cao, Hanqiang; Xu, Zijian

    2011-11-01

    Numerous edge detection methods have been proposed to detect image edges. However, these methods are not very effective in detecting edges in strong noisy images. Recent years, multiscale analysis has been introduced to the realm of image processing. As the third generation wavelet, shearlets have their own superiority. Anisotropic dilation operator and shear operator are introduced to overcome the shortcomings of traditional wavelets. Because of their sensitivity to directions, shearlets are apt to do the job of edge detection. Based on shearlets, in this paper, a new edge detection method is proposed. The main idea about this new method is combining the shearlet denoising method with the edge detecting method based on shearlets. Analyzing results show that edges are characterized as zerocrossing points in shearlet domain and can be extracted from shearlet transform coefficients by detecting zero crossing points and using boundary tracking method. Many experiments are conducted to test this novel approach and we also compare Sobel, Log and Canny operators with this new method. Experiments demonstrate that when an image existing high deviation Gaussian noise, this method are much better than ordinary edge detection operators in time domain.

  9. Ruthenium oxide modified nickel electrode for ascorbic acid detection.

    PubMed

    Lee, Yuan-Gee; Liao, Bo-Xuan; Weng, Yu-Ching

    2017-04-01

    Electrodes of ruthenium oxide modified nickel were prepared by a thermal decomposition method. The stoichiometry of the modifier, RuOx, was quantitatively determined to be a meta-stable phase, RuO5. The electrodes were employed to sense ascorbic acid in alkaline solution with a high sensitivity, 296 μAcm(-2) mM(-1), and good selectivity for eight kinds of disturbing reagents. We found that the ascorbic acid was oxidized irreversibly in solution. To match with the variation of the morphology, the sensitivity reached a maximum when the RuOx segregated with a nano-crystalline feature. We find that the substrate oxidized as the deposited RuOx grew thicker. The feature of the deposited RuOx changed from nano-particles to small islands resulting from the wetting effect of the substrate oxide, NiO; meanwhile the sensitivity decreased dramatically. The endurance of the RuOx/Ni electrode also showed a good performance after 38 days of successive test.

  10. Method for detection of antibodies for metallic elements

    DOEpatents

    Barrick, Charles W.; Clarke, Sara M.; Nordin, Carl W.

    1993-11-30

    An apparatus and method for detecting antibodies specific to non-protein antigens. The apparatus is an immunological plate containing a plurality of plastic projections coated with a non-protein material. Assays utilizing the plate are capable of stabilizing the non-protein antigens with detection levels for antibodies specific to the antigens on a nanogram level. A screening assay with the apparatus allows for early detection of exposure to non-protein materials. Specifically metallic elements are detected.

  11. Method for detection of antibodies for metallic elements

    DOEpatents

    Barrick, C.W.; Clarke, S.M.; Nordin, C.W.

    1993-11-30

    An apparatus and method for detecting antibodies specific to non-protein antigens. The apparatus is an immunological plate containing a plurality of plastic projections coated with a non-protein material. Assays utilizing the plate are capable of stabilizing the non-protein antigens with detection levels for antibodies specific to the antigens on a nanogram level. A screening assay with the apparatus allows for early detection of exposure to non-protein materials. Specifically metallic elements are detected. 10 figures.

  12. Method of Detecting Coliform Bacteria from Reflected Light

    NASA Technical Reports Server (NTRS)

    Vincent, Robert K. (Inventor)

    2014-01-01

    The present invention relates to a method of detecting coliform bacteria in water from reflected light, and also includes devices for the measurement, calculation and transmission of data relating to that method.

  13. Method and compositions for detecting of bloodstains using fluorescin-fluorescein reaction

    DOEpatents

    Di Benedetto, John; Kyle, Kevin; Boan, Terry; Marie, Charlene

    2004-02-17

    A method, compositions and kit are set forth for detecting blood stains. A reactant solution includes fluorescin solubilized (reduced) in acetic acid in ethanol. The solution may be buffered to a pH of approximately 9. After spraying the reactant solution on the suspected area an oxidizer is applied to promote the fluorescin to fluorescein reaction with the blood. The reacted fluorescein is then detected through luminescence for capture by photography.

  14. Heterodyne method for high specificity gas detection.

    NASA Technical Reports Server (NTRS)

    Dimeff, J.; Donaldson, R. W.; Gunter, W. D., Jr.; Jaynes, D. N.; Margozzi, A. P.; Deboo, G. J.; Mcclatchie, E. A.; Williams, K. G.

    1971-01-01

    This paper describes a new technique for measuring trace quantities of gases. The technique involves the use of a reference cell (containing a known amount of the gas being sought) and a sample cell (containing an unknown amount of the same gas) wherein the gas densities are modulated. Light passing through the two cells in sequence is modulated in intensity at the vibrational-rotational lines characteristic of the absorption spectrum for the gas of interest. Since the absorption process is nonlinear, modulating the two absorption cells at two different frequencies gives rise to a heterodyning effect, which in turn introduces sum and difference frequencies in the detected signal. Measuring the ratio of the difference frequency signal for example, to the signal introduced by the reference cell provides a normalized measure of the amount of the gas in the sample cell. The readings produced are thereby independent of source intensity, window transparency, and detector sensitivity. Experimental evaluation of the technique suggests that it should be applicable to a wide range of gases, that it should be able to reject spurious signals due to unwanted gases, and that it should be sensitive to concentrations of the order of 10 to the minus 8th power when used with a sample cell of only 20 cm length.

  15. Laser pulse detection method and apparatus

    NASA Technical Reports Server (NTRS)

    Goss, W.; Janesick, J. R. (Inventor)

    1984-01-01

    A sensor is described for detecting the difference in phase of a pair of returned light pulse components, such as two components of a light pulse of an optical gyro. In an optic gyro, the two light components have passed in opposite directions through a coil of optical fiber, with the difference in phase of the returned light components determining the intensity of light shining on the sensor. The sensor includes a CCD (charge coupled device) that receives the pair of returned light components to generate a charge proportional to the number of photons in the received light. The amount of the charge represents the phase difference between the two light components. At a time after the transmission of the light pulse and before the expected time of arrival of the interfering light components, charge accumulating in the CCD as a result of reflections from components in the system, are repeatedly removed from the CCD, by transferring out charges in the CCD and dumping these charges.

  16. Laser pulse detection method and apparatus

    NASA Technical Reports Server (NTRS)

    Goss, Willis C. (Inventor); Janesick, James R. (Inventor)

    1987-01-01

    A sensor is described for detecting the difference in phase of a pair of returned light pulse components, such as the two components of a light pulse of an optical gyro. In an optic gyro, the two light components have passed in opposite directions through a coil of optical fiber, with the difference in phase of the returned light components determining the intensity of light shining on the sensor. The sensor includes a CCD (charge coupled device) that receives the pair of returned light components to generate a charge proportional to the number of photons in the received light. The amount of the charge represents the phase difference between the two light components. At a time after the transmission of the light pulse and before the expected time of arrival of the interfering light components, charge accumulating in the CCD as a result of reflections from optical components in the system, are repeatedly removed from the CCD, by transferring out charges in the CCD and dumping these charges.

  17. Phenylboronic acid functionalized gold nanoparticles for highly sensitive detection of Staphylococcus aureus

    NASA Astrophysics Data System (ADS)

    Wang, Jine; Gao, Jingqing; Liu, Dianjun; Han, Dongxue; Wang, Zhenxin

    2012-01-01

    Herein, we report a phenylboronic acid functionalized gold nanoparticle (GNP)-based colorimetric assay for rapid detection of Staphylococcus aureus (S. aureus) with high sensitivity. In this approach, GNPs can bind to S. aureus by the reaction of phenylboronic acid with the cis-diol configuration in glycans on the bacterial surface, providing a colorimetric readout of the binding event. Using this strategy, we have been able to quantify S. aureus at a concentration of 50 cells per mL (three times the standard deviation divided by the slope of the working curve) in aqueous solution.Herein, we report a phenylboronic acid functionalized gold nanoparticle (GNP)-based colorimetric assay for rapid detection of Staphylococcus aureus (S. aureus) with high sensitivity. In this approach, GNPs can bind to S. aureus by the reaction of phenylboronic acid with the cis-diol configuration in glycans on the bacterial surface, providing a colorimetric readout of the binding event. Using this strategy, we have been able to quantify S. aureus at a concentration of 50 cells per mL (three times the standard deviation divided by the slope of the working curve) in aqueous solution. Electronic supplementary information (ESI) available: Details of experimental method and additional figures are available. See DOI: 10.1039/c2nr11657j

  18. Hypochlorite scavenging activity of hydroxycinnamic acids evaluated by a rapid microplate method based on the measurement of chloramines.

    PubMed

    Firuzi, Omidreza; Giansanti, Luisa; Vento, Roberta; Seibert, Cathrin; Petrucci, Rita; Marrosu, Giancarlo; Agostino, Roberta; Saso, Luciano

    2003-07-01

    Scavengers of hypochlorite (XOCl) could have beneficial effects in diseases in which this oxidant plays a pathogenic role. It has been reported that ferulic acid and chlorogenic acid, the quinic ester of caffeic acid, are good hypochlorite scavengers, but a systematic evaluation of the naturally occurring hydroxycinnamic acids (HCAs), which these substances belong to, has not been performed yet. Thus, in this work we studied, by two different in-vitro methods, the antioxidant activity of five HCAs: p-coumaric acid, ferulic acid, sinapinic acid, caffeic acid and chlorogenic acid. The methods applied in this study were based on the oxidation of human serum albumin (HSA) by XOCl, a new microplate method based on the measurement of chloramines and a previously described carbonyl assay. Firstly, lysine-derived chloramines, in the presence or absence of the HCAs, were detected using 5-thio-2-nitrobenzoic acid (TNB), measuring the absorbance at 415 nm by a microplate reader. To remove excess XOCl, Trolox, a known XOCl scavenger, was added before TNB. Secondly, lysine-derived carbonyls, in the presence or absence of the HCAs, were detected by using 2,4-dinitrophenylhydrazine. Hydroxycinnamic acids appeared active (caffeic >/= sinapinic > chlorogenic congruent with ferulic > p-coumaric acid) by both methods, suggesting possible pharmacological applications for these compounds, which are present at high concentrations in the plant kingdom.

  19. An HPLC method for the determination of selected amino acids in human embryo culture medium.

    PubMed

    Drábková, Petra; Andrlová, Lenka; Kanďár, Roman

    2017-02-01

    A method for the determination of selected amino acids in culture medium using HPLC with fluorescence detection is described. Twenty hours after intra-cytoplasmic sperm injection, one randomly selected zygote was transferred to the culture medium. After incubation (72 h after fertilization), the culture medium in which the embryo was incubated and blank medium was immediately stored at -80°C. Filtered medium samples were derivatized with ortho-phthalaldehyde (naphthalene-2,3-dicarboxaldehyde), forming highly fluorescent amino acids derivatives. Reverse-phase columns (LichroCART, Purospher STAR RP18e or Ascentis Express C18 ) were used for the separation. The derivatives were analyzed by gradient elution with a mobile phase containing ethanol and sodium dihydrogen phosphate. The analytical performance of this method is satisfactory for all amino acids; the intra-assay coefficients of variation were <10% and quantitative recoveries were between 95.5 and 104.4%. Changes in the levels of selected amino acids before and after human embryo cultivation were observed. After embryo incubation, the levels of all amino acids in the medium were increased, apart from aspartate and asparagine. After the cultivation of some embryos, amino acids which were not part of the medium were detected. Low amino acids turnover was observed in some embryos.

  20. Localized surface plasmon resonance mercury detection system and methods

    DOEpatents

    James, Jay; Lucas, Donald; Crosby, Jeffrey Scott; Koshland, Catherine P.

    2016-03-22

    A mercury detection system that includes a flow cell having a mercury sensor, a light source and a light detector is provided. The mercury sensor includes a transparent substrate and a submonolayer of mercury absorbing nanoparticles, e.g., gold nanoparticles, on a surface of the substrate. Methods of determining whether mercury is present in a sample using the mercury sensors are also provided. The subject mercury detection systems and methods find use in a variety of different applications, including mercury detecting applications.

  1. [Detection of erucic acid and glucosinolate in intact rapeseed by near-infrared diffuse reflectance spectroscopy].

    PubMed

    Riu, Yu-kui; Huang, Kun-lun; Wang, Wei-min; Guo, Jing; Jin, Yin-hua; Luo, Yun-bo

    2006-12-01

    With the rapid development of transgenic food, more and more transgenic food has been pouring into the market, raising great concern about transgenic food' s edible safety. To analyze the content of erucic acid and glucosinolate in transgenic rapeseed and its parents, all the seeds were scanned intact by continuous wave of near infrared diffuse reflectance spectrometry ranging from 12 000 to 4 000 cm(-1) with a resolution of 4 cm(-1) and 64 times of scanning. Bruker OPUS software package was applied for quantification, while the results were compared with the standard methods. The results showed that the method of NIRS was very precise, which proved that infrared diffuse reflectance spectroscopy can be applied to detect the toxins in transgenic food. On the other hand, the results also showed that the content of erucic acid in transgenic rapeseeds is 0. 5-1. 0 times

  2. Sensitive life detection: extraction of nucleic acids sorbing to Mars analogue minerals

    NASA Astrophysics Data System (ADS)

    Direito, S. O. L.; Marees, A.; Röling, W. F. M.

    2011-10-01

    The main goal of space missions to Mars is to find irrefutable proof of life. Consequently, the development, evaluation and optimization of sensitive extraction and detection methods for biomarkers are of extreme importance. Our aim consisted in the optimization of sensitive extraction techniques for molecules storing hereditary information (nucleic acids such as DNA), since these are common in life forms. However, adsorption of nucleic acids to mineral matrixes and soils can generate low extraction yields. Therefore, a second aim was to determine adsorption and identify 'problematic' Mars analogue minerals. In addition, the development of a method for quantification of DNA recovery by the use of an internal control was proved to be essential, since sensitive extraction needs information on recovery.

  3. Criteria For Evaluation of Proposed Protozoan Detection Methods

    EPA Science Inventory

    Currently, the only EPA approved method for detection and quantitation of protozoan cysts and oöcysts in source and drinking water, is the “ICR Protozoan Method for Detecting Giardia Cysts and Cryptosporidium Oöcysts in Water by a Fluorescent Antibody Procedure (ICR Microbial La...

  4. Ultra-high sensitivity radiation detection apparatus and method

    DOEpatents

    Gross, Kenneth C.; Valentine, John D.; Markum, Francis; Zawadzki, Mary; Dickerman, Charles

    1999-01-01

    A method and apparatus are provided to concentrate and detect very low levels of radioactive noble gases from the atmosphere. More specifically the invention provides a method and apparatus to concentrate xenon, krypton and radon in an organic fluid and to detect these gases by the radioactive emissions.

  5. Use of molecular methods for the detection of fungal spores.

    PubMed

    Ward, Elaine

    2009-01-01

    Traditional methods for the isolation and identification of fungal spores can be time-consuming and laborious. DNA-based methods for fungal detection can be used to detect the spores of plant-pathogenic fungi. Air borne spores can be collected and identified by PCR allowing identification of the species.

  6. Method for detecting trace impurities in gases

    DOEpatents

    Freund, Samuel M.; Maier, II, William B.; Holland, Redus F.; Beattie, Willard H.

    1981-01-01

    A technique for considerably improving the sensitivity and specificity of infrared spectrometry as applied to quantitative determination of trace impurities in various carrier or solvent gases is presented. A gas to be examined for impurities is liquefied and infrared absorption spectra of the liquid are obtained. Spectral simplification and number densities of impurities in the optical path are substantially higher than are obtainable in similar gas-phase analyses. Carbon dioxide impurity (.about.2 ppm) present in commercial Xe and ppm levels of Freon 12 and vinyl chloride added to liquefied air are used to illustrate the method.

  7. Method for detecting trace impurities in gases

    DOEpatents

    Freund, S.M.; Maier, W.B. II; Holland, R.F.; Beattie, W.H.

    A technique for considerably improving the sensitivity and specificity of infrared spectrometry as applied to quantitative determination of trace impurities in various carrier or solvent gases is presented. A gas to be examined for impurities is liquefied and infrared absorption spectra of the liquid are obtained. Spectral simplification and number densities of impurities in the optical path are substantially higher than are obtainable in similar gas-phase analyses. Carbon dioxide impurity (approx. 2 ppM) present in commercial Xe and ppM levels of Freon 12 and vinyl chloride added to liquefied air are used to illustrate the method.

  8. Evaluation of spectrophotometric and HPLC methods for shikimic acid determination in plants: models in glyphosate-resistant and -susceptible crops.

    PubMed

    Zelaya, Ian A; Anderson, Jennifer A H; Owen, Micheal D K; Landes, Reid D

    2011-03-23

    Endogenous shikimic acid determinations are routinely used to assess the efficacy of glyphosate in plants. Numerous analytical methods exist in the public domain for the detection of shikimic acid, yet the most commonly cited comprise spectrophotometric and high-pressure liquid chromatography (HPLC) methods. This paper compares an HPLC and two spectrophotometric methods (Spec 1 and Spec 2) and assesses the effectiveness in the detection of shikimic acid in the tissues of glyphosate-treated plants. Furthermore, the study evaluates the versatility of two acid-based shikimic acid extraction methods and assesses the longevity of plant extract samples under different storage conditions. Finally, Spec 1 and Spec 2 are further characterized with respect to (1) the capacity to discern between shikimic acid and chemically related alicyclic hydroxy acids, (2) the stability of the chromophore (t1/2), (3) the detection limits, and (4) the cost and simplicity of undertaking the analytical procedure. Overall, spectrophotometric methods were more cost-effective and simpler to execute yet provided a narrower detection limit compared to HPLC. All three methods were specific to shikimic acid and detected the compound in the tissues of glyphosate-susceptible crops, increasing exponentially in concentration within 24 h of glyphosate application and plateauing at approximately 72 h. Spec 1 estimated more shikimic acid in identical plant extract samples compared to Spec 2 and, likewise, HPLC detection was more effective than spectrophotometric determinations. Given the unprecedented global adoption of glyphosate-resistant crops and concomitant use of glyphosate, an effective and accurate assessment of glyphosate efficacy is important. Endogenous shikimic acid determinations are instrumental in corroborating the efficacy of glyphosate and therefore have numerous applications in herbicide research and related areas of science as well as resolving many commercial issues as a consequence of

  9. Delamination detection using methods of computational intelligence

    NASA Astrophysics Data System (ADS)

    Ihesiulor, Obinna K.; Shankar, Krishna; Zhang, Zhifang; Ray, Tapabrata

    2012-11-01

    Abstract Reliable delamination prediction scheme is indispensable in order to prevent potential risks of catastrophic failures in composite structures. The existence of delaminations changes the vibration characteristics of composite laminates and hence such indicators can be used to quantify the health characteristics of laminates. An approach for online health monitoring of in-service composite laminates is presented in this paper that relies on methods based on computational intelligence. Typical changes in the observed vibration characteristics (i.e. change in natural frequencies) are considered as inputs to identify the existence, location and magnitude of delaminations. The performance of the proposed approach is demonstrated using numerical models of composite laminates. Since this identification problem essentially involves the solution of an optimization problem, the use of finite element (FE) methods as the underlying tool for analysis turns out to be computationally expensive. A surrogate assisted optimization approach is hence introduced to contain the computational time within affordable limits. An artificial neural network (ANN) model with Bayesian regularization is used as the underlying approximation scheme while an improved rate of convergence is achieved using a memetic algorithm. However, building of ANN surrogate models usually requires large training datasets. K-means clustering is effectively employed to reduce the size of datasets. ANN is also used via inverse modeling to determine the position, size and location of delaminations using changes in measured natural frequencies. The results clearly highlight the efficiency and the robustness of the approach.

  10. Vapor generation methods for explosives detection research

    SciTech Connect

    Grate, Jay W.; Ewing, Robert G.; Atkinson, David A.

    2012-12-01

    The generation of calibrated vapor samples of explosives compounds remains a challenge due to the low vapor pressures of the explosives, adsorption of explosives on container and tubing walls, and the requirement to manage (typically) multiple temperature zones as the vapor is generated, diluted, and delivered. Methods that have been described to generate vapors can be classified as continuous or pulsed flow vapor generators. Vapor sources for continuous flow generators are typically explosives compounds supported on a solid support, or compounds contained in a permeation or diffusion device. Sources are held at elevated isothermal temperatures. Similar sources can be used for pulsed vapor generators; however, pulsed systems may also use injection of solutions onto heated surfaces with generation of both solvent and explosives vapors, transient peaks from a gas chromatograph, or vapors generated by s programmed thermal desorption. This article reviews vapor generator approaches with emphasis on the method of generating the vapors and on practical aspects of vapor dilution and handling. In addition, a gas chromatographic system with two ovens that is configurable with up to four heating ropes is proposed that could serve as a single integrated platform for explosives vapor generation and device testing. Issues related to standards, calibration, and safety are also discussed.

  11. Voltammetric iodometric titration of ascorbic acid with dead-stop end-point detection in fresh vegetables and fruit samples.

    PubMed

    Verdini, R A; Lagier, C M

    2000-07-01

    The present work describes a method for determining ascorbic acid, which combines iodometry with a voltammetric technique to detect the end point of the titration. In addition, the validity of the method applied to natural vegetable or fruit samples was assessed. The results were compared with those obtained by an accurate method such as HPLC using UV detection. Similar values of ascorbic acid for different natural samples were obtained by means of this approach (p > 0.05). The limit of quantification was 0.1 mg. This technique presents the advantage of other electroanalytical methods such as avoiding filtration or ultracentrifugation steps, with the additional benefit of using the platinum electrodes, which are routinely used in the laboratory. These facts allow a rapid and efficient quantification of ascorbic acid with very low cost of reagents and equipment.

  12. Detection of acid moisture in photovoltaic modules using a dual wavelength pH-sensitive fluorescent dye

    NASA Astrophysics Data System (ADS)

    Asaka, Takashi; Iwami, Kentaro; Taguchi, Atsushi; Umeda, Norihiro; Masuda, Atsushi

    2014-01-01

    The formation of acetic acid via the penetration of moisture into ethylene vinyl acetate (EVA) in photovoltaic (PV) modules is cited as the main reason for PV modules’ degradation. Currently, there is no effective method for detecting acetic moisture in PV modules. We proposed a simple method for detecting acid moisture in PV modules using a dual-wavelength pH-sensitive dye that measures pH by the ratio of the intensities of two peaks in the fluorescence spectra of the dye. We detected the pH change caused by acetic acid with the change in the intensity ratio of the fluorescence spectra of the dried dye. Furthermore, we observed that the dry fluorescent dye is heat resistant to withstand the lamination process for the manufacturing of PV modules, and has good long-term durability.

  13. Method And Apparatus For Detecting Chemical Binding

    DOEpatents

    Warner, Benjamin P.; Havrilla, George J.; Miller, Thomasin C.; Wells, Cyndi A.

    2005-02-22

    The method for screening binding between a target binder and potential pharmaceutical chemicals involves sending a solution (preferably an aqueous solution) of the target binder through a conduit to a size exclusion filter, the target binder being too large to pass through the size exclusion filter, and then sending a solution of one or more potential pharmaceutical chemicals (preferably an aqueous solution) through the same conduit to the size exclusion filter after target binder has collected on the filter. The potential pharmaceutical chemicals are small enough to pass through the filter. Afterwards, x-rays are sent from an x-ray source to the size exclusion filter, and if the potential pharmaceutical chemicals form a complex with the target binder, the complex produces an x-ray fluorescence signal having an intensity that indicates that a complex has formed.

  14. Method and apparatus for detecting chemical binding

    DOEpatents

    Warner, Benjamin P.; Havrilla, George J.; Miller, Thomasin C.; Wells, Cyndi A.

    2007-07-10

    The method for screening binding between a target binder and potential pharmaceutical chemicals involves sending a solution (preferably an aqueous solution) of the target binder through a conduit to a size exclusion filter, the target binder being too large to pass through the size exclusion filter, and then sending a solution of one or more potential pharmaceutical chemicals (preferably an aqueous solution) through the same conduit to the size exclusion filter after target binder has collected on the filter. The potential pharmaceutical chemicals are small enough to pass through the filter. Afterwards, x-rays are sent from an x-ray source to the size exclusion filter, and if the potential pharmaceutical chemicals form a complex with the target binder, the complex produces an x-ray fluorescence signal having an intensity that indicates that a complex has formed.

  15. A fully validated GC-TOF-MS method for the quantification of fatty acids revealed alterations in the metabolic profile of fatty acids after smoking cessation.

    PubMed

    Goettel, Michael; Niessner, Reinhard; Pluym, Nikola; Scherer, Gerhard; Scherer, Max

    2017-01-15

    We developed and validated an efficient and robust method for the simultaneous quantification of 44 fatty acid species in human plasma via GC-TOF-MS. The method is characterized by its robustness, accuracy and precision covering a wide range of fatty acid species with various saturation degrees including short chain fatty acids (beginning with FA 4:0) and long chain fatty acids (up to FA 32:0). The fatty acids were methylated prior to analyses and subsequently detected as fatty acid methyl esters by means of GC-TOF-MS. A highly substituted polar column allowed the separation of geometrical and positional isomers of fatty acid species. The method was applied to plasma samples of a strictly diet controlled clinical smoking cessation study including 39 smokers followed over the course of three months after having quit. Statistical significant alterations within the fatty acid profile were observed when comparing the baseline (subjects still smoking) with one week, one month and three months of smoking cessation. After 3 months of smoking cessation, a partial recovery of alterations in the fatty acid profile evoked by smoking was observed. In conclusion, the developed fatty acid profiling method using GC-TOF-MS has proven as a reliable tool for the quantitative determination of 44 individual fatty acid species within clinical studies.

  16. Comparison of multi-mode parallel detection microscopy methods

    NASA Astrophysics Data System (ADS)

    Zhu, Dazhao; Fang, Yue; Chen, Youhua; Hussain, Anwar; Kuang, Cuifang; Ding, Zhihua; Liu, Xu

    2017-03-01

    Four microscopy resolution enhancement methods based on parallel detection were investigated in this study: confocal microscopy with four pinhole sizes, fluorescence emission difference microscopy (FED) based on parallel detection, Airyscan microscopy, and virtual k-vector modulation optical microscopy (Vikmom). These methods use different algorithms to process parallel detection data and achieve resolution improvement. We investigated these methods first by performing simulations and then experimentally. In this report, the basic theories of these methods are briefly introduced. Then, analyses and comparisons of their imaging performances, especially in terms of resolution improvement, imaging speed, and signal-to-noise ratio, are presented. Finally, the results of our comparative study are summarized.

  17. Reliably Detectable Flaw Size for NDE Methods that Use Calibration

    NASA Technical Reports Server (NTRS)

    Koshti, Ajay M.

    2017-01-01

    Probability of detection (POD) analysis is used in assessing reliably detectable flaw size in nondestructive evaluation (NDE). MIL-HDBK-1823 and associated mh1823 POD software gives most common methods of POD analysis. In this paper, POD analysis is applied to an NDE method, such as eddy current testing, where calibration is used. NDE calibration standards have known size artificial flaws such as electro-discharge machined (EDM) notches and flat bottom hole (FBH) reflectors which are used to set instrument sensitivity for detection of real flaws. Real flaws such as cracks and crack-like flaws are desired to be detected using these NDE methods. A reliably detectable crack size is required for safe life analysis of fracture critical parts. Therefore, it is important to correlate signal responses from real flaws with signal responses form artificial flaws used in calibration process to determine reliably detectable flaw size.

  18. Method for enhancing amidohydrolase activity of fatty acid amide hydrolase

    SciTech Connect

    John, George; Nagarajan, Subbiah; Chapman, Kent; Faure, Lionel; Koulen, Peter

    2016-10-25

    A method for enhancing amidohydrolase activity of Fatty Acid Amide Hydrolase (FAAH) is disclosed. The method comprising administering a phenoxyacylethanolamide that causes the enhanced activity. The enhanced activity can have numerous effects on biological organisms including, for example, enhancing the growth of certain seedlings. The subject matter disclosed herein relates to enhancers of amidohydrolase activity.

  19. Method to produce succinic acid from raw hydrolysates

    DOEpatents

    Donnelly, Mark I.; Sanville-Millard, Cynthia Y.; Nghiem, Nhuan Phu

    2004-06-01

    A method for producing succinic acid from industrial-grade hydrolysates is provided, comprising supplying an organism that contains mutations for the genes ptsG, pflB, and ldhA, allowing said organism to accumulate biomass, and allowing said organism to metabolize the hydrolysate. Also provided is a bacteria mutant characterized in that it produces succinic acid from substrate contained in industrial-grade hydrolysate in a ratio of between 0.6:1 and 1.3:1 succinic acid to substrate.

  20. Application of a bioluminescence method for the beer industry: sensitivity of MicroStar-RMDS for detecting beer-spoilage bacteria. Rapid Microbe Detection System.

    PubMed

    Takahashi, T; Nakakita, Y; Watari, J; Shinotsuka, K

    2000-05-01

    We set up the original operating conditions of the MicroStar-Rapid Microbe Detection System (RMDS) to suppress false positives, which have kept this system from practical. The detection limit of our system was between 6.3 x 10(-16) mol and 3.1 x 10(-16) mol in terms of the amount of ATP, which is approximately equal to the ATP content of one yeast cell or 50 lactic acid bacteria cells. The detection time and the detection count were compared between the RMD method and the conventional plate count method (C.P.C. method) using 23 test samples of beer-spoilage Lactobacillus brevis. Judging from the detection time and detection count, 16-24 hours of cultivation for the RMD method corresponded to 40-96 hours of cultivation for the C.P.C. method. The RMD method reached a useful level for our practical use at the point of sensitivity.

  1. Simultaneous determination of acetylsalicylic acid and salicylic acid in human plasma by isocratic high-pressure liquid chromatography with post-column hydrolysis and fluorescence detection.

    PubMed

    Hobl, Eva-Luise; Jilma, Bernd; Ebner, Josef; Schmid, Rainer W

    2013-06-01

    A selective, sensitive and rapid high-performance liquid chromatography method with post-column hydrolysis and fluorescence detection was developed for the simultaneous quantification of acetylsalicylic acid and its metabolite salicylic acid in human plasma. Following the addition of 2-hydroxy-3-methoxybenzoic acid as internal standard and simple protein precipitation with acetonitrile, the analytes were separated on a ProntoSIL 120 C18 ace-EPS column (150 × 2 mm, 3 µm) protected by a C8 guard column (5 µm). The mobile phase, 10 mm formic acid in water (pH 2.9) and acetonitrile (70:30, v/v), was used at a flow rate of 0.35 mL/min. After on-line post-column hydrolysis of acetylsalicylic acid (ASA) to salicylic acid (SA) by addition of alkaline solution, the analytes were measured at 290 nm (λex ) and 400 nm (λem ). The method was linear in the concentration ranges between 0.05 and 20 ng/μL for both ASA and SA with a lower limit of quantification of 25 pg/μL for SA and 50 pg/μL for ASA. The limit of detection was 15 pg/μL for SA and 32.5 pg/μL for ASA. The analysis of ASA and SA can be carried out within 8 min; therefore this method is suitable for measuring plasma concentrations of salicylates in clinical routine.

  2. Determination of organic acids by CE and CEC methods.

    PubMed

    Klampfl, Christian W

    2007-10-01

    A comprehensive overview of the analysis of low-molecular-mass organic acids employing electromigration methods in the capillary format is given. This review includes papers published since 2003 and can be seen as an update of the review paper published by Galli et al. in 2003. Tables included in this review contain application papers describing the determination of organic acids from a variety of fields like the analysis of food and beverages, environmental samples, samples from clinical origin, and from natural products.

  3. Simultaneous analysis of six aristolochic acids and five aristolactams in herbal plants and their preparations by high-performance liquid chromatography-diode array detection-fluorescence detection.

    PubMed

    Yuan, Jinbin; Liu, Qian; Zhu, Weifeng; Ding, Li; Tang, Fei; Yao, Shouzhuo

    2008-02-22

    Aristolochic acid analogues, including aristolochic acids (AAs) and aristolactams (ALs), are known to be nephrotoxic, carcinogenic and mutagenic. In this paper, a high-performance liquid chromatography-diode array detection-fluorescence detection (HPLC-DAD-FLD) method was developed for the simultaneous determination of six AAs together with five ALs. Baseline separation was obtained on an ODS C18 analytical column with 0.2% HAc/methanol gradient elution. The hyphenation of DAD and FLD allows the method to directly meet the analysis requirements of most herbal plants with high sensitivity and selectivity. For trace analysis, aristolochic acids were reduced to their corresponding aritstolactams in acidic solution containing iron powder, and then high sensitive detection and quantification were carried out. The method was successfully validated in the matrices of various Aristolochiaceae plants and their preparations. Linearities of around 3-4 orders of magnitude were obtained with correlation coefficients exceeding 0.9970. The detection limits were decreased to 0.2ng/ml. Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 5.74%, and the average recovery factors were in the range of 94.5-99.2%.

  4. Zinc oxide nanowires-based electrochemical biosensor for L-lactic acid amperometric detection

    NASA Astrophysics Data System (ADS)

    Zhao, Yanguang; Yan, Xiaoqin; Kang, Zhuo; Fang, Xiaofei; Zheng, Xin; Zhao, Lanqing; Du, Hongwu; Zhang, Yue

    2014-05-01

    In this work, zinc oxide (ZnO) nanowires-based electrochemical biosensor is designed and fabricated for the detection of L-lactic acid. ZnO nanowires were successfully synthesized via the chemical vapor deposition method. The morphology and structure of the prepared products were characterized, and the average diameter of synthesized ZnO samples was 500 nm. The fluorescence characterization was performed to verify the immobilization of lactate oxidase onto the ZnO surface. Biosensors based on large-area ZnO nanowires were then constructed, and a series of electrochemical experiments showed that ZnO could provide the efficient electron transfer channel between the enzymic active sites and the electrode surface. The proposed electrochemical biosensor exhibited a sensitivity of 15.6 µA cm-2 mM-1, a wide linear range of 12 µM-1.2 mM with a low-detection limit of 12 µM for L-lactic acid detection. This study has indicated the potential applications for ZnO nanowires to construct the simple and economic nano-bio devices for the detection of biological species.

  5. Effects of Linking Methods on Detection of DIF.

    ERIC Educational Resources Information Center

    Kim, Seock-Ho; Cohen, Allan S.

    1992-01-01

    Effects of the following methods for linking metrics on detection of differential item functioning (DIF) were compared: (1) test characteristic curve method (TCC); (2) weighted mean and sigma method; and (3) minimum chi-square method. With large samples, results were essentially the same. With small samples, TCC was most accurate. (SLD)

  6. [Methods for detection of methylated cytosine residues in DNA].

    PubMed

    Smirnikhina, S A; Lavrov, A V

    2009-01-01

    The article provides analysis of common methods for DNA methylation detection. Advantages and limitations of methods used for different purposes are compared. Clue step for most common methods is bisulfite treatment of DNA samples and its protocol is described in details. Recommendations are formulated for each method best in solving specific problems.

  7. Systems and Methods for Automated Water Detection Using Visible Sensors

    NASA Technical Reports Server (NTRS)

    Rankin, Arturo L. (Inventor); Matthies, Larry H. (Inventor); Bellutta, Paolo (Inventor)

    2016-01-01

    Systems and methods are disclosed that include automated machine vision that can utilize images of scenes captured by a 3D imaging system configured to image light within the visible light spectrum to detect water. One embodiment includes autonomously detecting water bodies within a scene including capturing at least one 3D image of a scene using a sensor system configured to detect visible light and to measure distance from points within the scene to the sensor system, and detecting water within the scene using a processor configured to detect regions within each of the at least one 3D images that possess at least one characteristic indicative of the presence of water.

  8. Amplified detection of nucleic acid by G-quadruplex based hybridization chain reaction.

    PubMed

    Dong, Juan; Cui, Xin; Deng, Yun; Tang, Zhuo

    2012-01-01

    A protein-free, isothermal, self-amplified nucleic acid sensing system which was a G-quadruplex integrated hybridization chain reaction (GQ-HCR) system was developed. The G-quadruplex was closed two-thirds in the loop and one-third in the stem of one of the GQ-HCR hairpin probes. In the absence of the target molecule, the GQ-HCR probes stayed as inactive meta-stable hairpin structures and the G-quadruplex was inert. Reversely, the GQ-HCR probes could be cross-opened to start a hybridization chain reaction and the closed G-quadruplex could be released to be free when the GQ-HCR probes came across the target molecule. The GQ-HCR nucleic acid sensing system could detect as low as 7.5 nM ssDNA or RNA by the colorimetric method and 4 nM ssDNA by the fluorometric method. Less than 10 copies of dsDNA template could also be detected when PCR was combined with the GQ-HCR system (PCR+GQ-HCR). Because of these advantages, the GQ-HCR system was also studied for application in visual chip detection to obtain a satisfactory repeatable and specific result.

  9. Microemulsion electrokinetic chromatography with laser-induced fluorescence detection: as tested with amino acid derivatives.

    PubMed

    Jianping, Xie; Jiyou, Zhang; Huanxiang, Liu; Jiaqin, Liu; Jianniao, Tian; Xingguo, Chen; Zhide, Hu

    2004-10-01

    Over a decade ago, microemulsion electrokinetic chromatography was introduced as a novel mode of capillary electrophoresis. However, there has not been publication on the combination of microemulsion electrokinetic chromatography with laser-induced fluorescence detection. In this paper, a preliminary method using microemulsion eletrokinetic chromatography combined with laser-induced fluorescence detection and second derivative electrophoregram was established as a sensitive and selective assay for separation and determination of nine amino acids after derivatization with 4-chloro-7-nitrobenzo-2-oxa-1, 3-diazol. The derivatization and separation conditions were optimized. In the investigated concentration ranges correlation coefficients were better than 0.995. The relative standard deviation (n = 5) of the migration times and peak heights were 0.56-0.76 and 2.21-7.15%, respectively. The detection limits (S/N = 3) were at a neaomolar level (0.32-2.20 nM). The method was applied for the analysis of compound amino acid injection and a Chinese traditional herbal medicine. The recoveries were 95.9-107.9%.

  10. Round-robin comparison of methods for the detection of human enteric viruses in lettuce.

    PubMed

    Le Guyader, Françoise S; Schultz, Anna-Charlotte; Haugarreau, Larissa; Croci, Luciana; Maunula, Leena; Duizer, Erwin; Lodder-Verschoor, Froukje; von Bonsdorff, Carl-Henrik; Suffredini, Elizabetha; van der Poel, Wim M M; Reymundo, Rosanna; Koopmans, Marion

    2004-10-01

    Five methods that detect human enteric virus contamination in lettuce were compared. To mimic multiple contaminations as observed after sewage contamination, artificial contamination was with human calicivirus and poliovirus and animal calicivirus strains at different concentrations. Nucleic acid extractions were done at the same time in the same laboratory to reduce assay-to-assay variability. Results showed that the two critical steps are the washing step and removal of inhibitors. The more reliable methods (sensitivity, simplicity, low cost) included an elution/concentration step and a commercial kit. Such development of sensitive methods for viral detection in foods other than shellfish is important to improve food safety.

  11. Determination of uric acid in human urine and serum by capillary electrophoresis with chemiluminescence detection.

    PubMed

    Zhao, Shulin; Wang, Jianshi; Ye, Fanggui; Liu, Yi-Ming

    2008-07-15

    A simple and sensitive method based on capillary electrophoresis (CE) with chemiluminescence (CL) detection has been developed for the determination of uric acid (UA). The sensitive detection was based on the enhancement effect of UA on the CL reaction between luminol and potassium ferricyanide (K3[Fe(CN)6]) in alkaline solution. A laboratory-built reaction flow cell and a photon counter were deployed for the CL detection. Experimental conditions for CL detection were studied in detail to achieve a maximum assay sensitivity. Optimal conditions were found to be 1.0 x 10(-4) M luminol added to the CE running buffer and 1.0 x 10(-4) M K3[Fe(CN)6] in 0.2 M NaOH solution introduced postcolumn. The proposed CE-CL assay showed good repeatability (relative standard deviation [RSD]=3.5%, n=11) and a detection limit of 3.5 x 10(-7) M UA (signal/noise ratio [S/N]=3). A linear calibration curve ranging from 6.0 x 10(-7) to 3.0 x 10(-5) M UA was obtained. The method was evaluated by quantifying UA in human urine and serum samples with satisfactory assay results.

  12. Harmonic Motion Microwave Doppler Imaging method for breast tumor detection.

    PubMed

    Top, Can Barıs; Tafreshi, Azadeh Kamali; Gençer, Nevzat G

    2014-01-01

    Harmonic Motion Microwave Doppler Imaging (HMMDI) method is recently proposed as a non-invasive hybrid breast imaging technique for tumor detection. The acquired data depend on acoustic, elastic and electromagnetic properties of the tissue. The potential of the method is analyzed with simulation studies and phantom experiments. In this paper, the results of these studies are summarized. It is shown that HMMDI method has a potential to detect malignancies inside fibro-glandular tissue.

  13. Nucleic acid amplification: Alternative methods of polymerase chain reaction

    PubMed Central

    Fakruddin, Md; Mannan, Khanjada Shahnewaj Bin; Chowdhury, Abhijit; Mazumdar, Reaz Mohammad; Hossain, Md. Nur; Islam, Sumaiya; Chowdhury, Md. Alimuddin

    2013-01-01

    Nucleic acid amplification is a valuable molecular tool not only in basic research but also in application oriented fields, such as clinical medicine development, infectious diseases diagnosis, gene cloning and industrial quality control. A comperehensive review of the literature on the principles, applications, challenges and prospects of different alternative methods of polymerase chain reaction (PCR) was performed. PCR was the first nucleic acid amplification method. With the advancement of research, a no of alternative nucleic acid amplification methods has been developed such as loop mediated isothermal amplification, nucleic acid sequence based amplification, strand displacement amplification, multiple displacement amplification. Most of the alternative methods are isothermal obviating the need for thermal cyclers. Though principles of most of the alternate methods are relatively complex than that of PCR, they offer better applicability and sensitivity in cases where PCR has limitations. Most of the alternate methods still have to prove themselves through extensive validation studies and are not available in commercial form; they pose the potentiality to be used as replacements of PCR. Continuous research is going on in different parts of the world to make these methods viable technically and economically. PMID:24302831

  14. Nucleic acid amplification: Alternative methods of polymerase chain reaction.

    PubMed

    Fakruddin, Md; Mannan, Khanjada Shahnewaj Bin; Chowdhury, Abhijit; Mazumdar, Reaz Mohammad; Hossain, Md Nur; Islam, Sumaiya; Chowdhury, Md Alimuddin

    2013-10-01

    Nucleic acid amplification is a valuable molecular tool not only in basic research but also in application oriented fields, such as clinical medicine development, infectious diseases diagnosis, gene cloning and industrial quality control. A comperehensive review of the literature on the principles, applications, challenges and prospects of different alternative methods of polymerase chain reaction (PCR) was performed. PCR was the first nucleic acid amplification method. With the advancement of research, a no of alternative nucleic acid amplification methods has been developed such as loop mediated isothermal amplification, nucleic acid sequence based amplification, strand displacement amplification, multiple displacement amplification. Most of the alternative methods are isothermal obviating the need for thermal cyclers. Though principles of most of the alternate methods are relatively complex than that of PCR, they offer better applicability and sensitivity in cases where PCR has limitations. Most of the alternate methods still have to prove themselves through extensive validation studies and are not available in commercial form; they pose the potentiality to be used as replacements of PCR. Continuous research is going on in different parts of the world to make these methods viable technically and economically.

  15. Evaluation of acidity estimation methods for mine drainage, Pennsylvania, USA.

    PubMed

    Park, Daeryong; Park, Byungtae; Mendinsky, Justin J; Paksuchon, Benjaphon; Suhataikul, Ratda; Dempsey, Brian A; Cho, Yunchul

    2015-01-01

    Eighteen sites impacted by abandoned mine drainage (AMD) in Pennsylvania were sampled and measured for pH, acidity, alkalinity, metal ions, and sulfate. This study compared the accuracy of four acidity calculation methods with measured hot peroxide acidity and identified the most accurate calculation method for each site as a function of pH and sulfate concentration. Method E1 was the sum of proton and acidity based on total metal concentrations; method E2 added alkalinity; method E3 also accounted for aluminum speciation and temperature effects; and method E4 accounted for sulfate speciation. To evaluate errors between measured and predicted acidity, the Nash-Sutcliffe efficiency (NSE), the coefficient of determination (R (2)), and the root mean square error to standard deviation ratio (RSR) methods were applied. The error evaluation results show that E1, E2, E3, and E4 sites were most accurate at 0, 9, 4, and 5 of the sites, respectively. Sites where E2 was most accurate had pH greater than 4.0 and less than 400 mg/L of sulfate. Sites where E3 was most accurate had pH greater than 4.0 and sulfate greater than 400 mg/L with two exceptions. Sites where E4 was most accurate had pH less than 4.0 and more than 400 mg/L sulfate with one exception. The results indicate that acidity in AMD-affected streams can be accurately predicted by using pH, alkalinity, sulfate, Fe(II), Mn(II), and Al(III) concentrations in one or more of the identified equations, and that the appropriate equation for prediction can be selected based on pH and sulfate concentration.

  16. The evaluation method for antiplatelet effect of acetylsalicylic acid.

    PubMed

    Yokoyama, Haruko; Mastumura, Takashi; Soeda, Shinji; Suzuki, Yuji; Watanabe, Masayuki; Kashiwakura, Emiko; Saso, Takayuki; Ikeda, Noriyuki; Tokuoka, Kentaro; Kitagawa, Yasuhisa; Yamada, Yasuhiko

    2014-12-01

    Reduced platelet aggregation by acetylsalicylic acid administration has been associated with adverse outcomes in patients with thrombotic diseases, thus it is important to determine aspirin resistance in those cases. The antiplatelet effect of acetylsalicylic acid is rarely measured, but it has many problems. The aim of this study was to find the evaluation method for antiplatelet effect after administration of acetylsalicylic acid. We developed a particle counting method based upon laser light scattering, and utilized the platelet aggregation agonists, collagen, at 0.25, 0.5 and 1.0 μg/mL, and adenosine diphosphate (ADP), at 0.5, 1.0 and 2.0 μM, to determine their effective concentrations. Seventeen healthy volunteers were administered acetylsalicylic acid at 162 mg/day, with platelet aggregation determined before and 20 min after administration. In all subjects, the rate of platelet aggregation induced by 1.0 μg/mL of collagen before taking acetylsalicylic acid was the highest value obtained, while 20 min after acetylsalicylic acid administration, aggregation induced by collagen at 1.0 μg/mL was significantly decreased as compared to before administration. As for the other concentrations of collagen and all those of ADP tested, platelet aggregation was either not significantly induced before taking acetylsalicylic acid or the rate of aggregation was not significantly decreased after taking acetylsalicylic acid. Our results indicate that collagen at 1.0 μg/mL is appropriate as a platelet aggregation agonist for evaluating the antiplatelet effect of acetylsalicylic acid. Thus, it is useful that the measurement is performed only once.

  17. A novel multi-object detection method in HSV space

    NASA Astrophysics Data System (ADS)

    Tang, Mingfeng; Li, Hongsong; Wen, Yane; Tang, Liping

    2013-07-01

    In order to solve the adverse effects of strong light and shadow on the test results, a fusion frame difference and background subtraction method in the HSV space is used in this paper. By using frame difference method to solve the effect of strong light, but frame difference method can not detect object when the object do not move, the method of background subtraction can detect it, building Gaussian background model in the HSV space can eliminate shadows. Empirical results show that the method of fusion frame difference and background subtraction in the HSV space can get overcome the effect of strong light and shadows. Fusion background subtraction and frame difference method based on establishing a Gaussian mixture model in HSV space can overcome the disadvantages of the frame difference method, at the same time it can also solve the false detection of object which result from the background subtraction method.

  18. Linking autotrophic activity in environmental samples with specific bacterial taxa by detection of 13C-labelled fatty acids.

    PubMed

    Knief, Claudia; Altendorf, Karlheinz; Lipski, André

    2003-11-01

    A method for the detection of physiologically active autotrophic bacteria in complex microbial communities was developed based on labelling with the stable isotope 13C. Labelling of autotrophic nitrifying, sulphur-oxidizing and iron-oxidizing populations was performed in situ by incubation with NaH[13C]O3. Incorporated label into fatty acid methyl esters (FAMEs) was detected and quantified using gas chromatography-mass spectrometry in single ion monitoring mode. Before the analyses of different environmental samples, the protocol was evaluated in pure culture experiments. In different environmental samples a selective labelling of fatty acids demonstrated which microbial taxa were responsible for the respective chemolithoautotrophic activity. The most strongly labelled fatty acids of a sample from a sulphide treating biofilter from an animal rendering plant were cis-7-hexadecenoic acid (16:1 cis7) and 11-methyl hexadecanoic acid (16:0 11methyl), which are as-yet not known for any sulphide-oxidizing autotroph. The fatty acid labelling pattern of an experimental biotrickling filter sample supplied with dimethyl disulphide clearly indicated the presence and activity of sulphide-oxidizing bacteria of the genus Thiobacillus. For a third environmental sample from an acid mining lake sediment, the assignment of autotrophic activity to bacteria of the genus Leptospirillum but not to Acidithiobacillus could be made by this method, as the fatty acid patterns of these bacteria show clear differences.

  19. 25 Years of Self-organized Criticality: Numerical Detection Methods

    NASA Astrophysics Data System (ADS)

    McAteer, R. T. James; Aschwanden, Markus J.; Dimitropoulou, Michaila; Georgoulis, Manolis K.; Pruessner, Gunnar; Morales, Laura; Ireland, Jack; Abramenko, Valentyna

    2016-01-01

    The detection and characterization of self-organized criticality (SOC), in both real and simulated data, has undergone many significant revisions over the past 25 years. The explosive advances in the many numerical methods available for detecting, discriminating, and ultimately testing, SOC have played a critical role in developing our understanding of how systems experience and exhibit SOC. In this article, methods of detecting SOC are reviewed; from correlations to complexity to critical quantities. A description of the basic autocorrelation method leads into a detailed analysis of application-oriented methods developed in the last 25 years. In the second half of this manuscript space-based, time-based and spatial-temporal methods are reviewed and the prevalence of power laws in nature is described, with an emphasis on event detection and characterization. The search for numerical methods to clearly and unambiguously detect SOC in data often leads us outside the comfort zone of our own disciplines—the answers to these questions are often obtained by studying the advances made in other fields of study. In addition, numerical detection methods often provide the optimum link between simulations and experiments in scientific research. We seek to explore this boundary where the rubber meets the road, to review this expanding field of research of numerical detection of SOC systems over the past 25 years, and to iterate forwards so as to provide some foresight and guidance into developing breakthroughs in this subject over the next quarter of a century.

  20. A RAPID METHOD FOR THE EXTRACTION OF FUNGAL DNA FROM ENVIRONMENTAL SAMPLES: EVALUATION IN THE QUANTITATIVE ANALYSIS OF MEMNONIELLA ECHINATA CONIDIA USING REAL TIME DETECTION OF PCR PRODUCTS

    EPA Science Inventory

    New technologies are creating the potential for using nucleic acid sequence detection to perform routine microbiological analyses of environmental samples. Our laboratory has recently reported on the development of a method for the quantitative detection of Stachybotrys chartarum...

  1. Lyophilized Visually Readable Loop-Mediated Isothermal Reverse Transcriptase Nucleic Acid Amplification Test for Detection Ebola Zaire RNA.

    PubMed

    Carter, Christoph; Akrami, Kevan; Hall, Drew; Smith, Davey; Aronoff-Spencer, Eliah

    2017-02-24

    Recent viral outbreaks highlight the need for reliable, yet broadly deployable diagnostics for detection of epidemic and emerging pathogens. In this study we designed and optimized methods to visually detect viral nucleic acid by isothermal amplification and SYBR dye intercalation. We designed and tested loop-mediated isothermal amplification (LAMP) primers and lyophilized reactions to optimize the detection of Zaire Ebola Virus (ZEBOV) and further evolved the LAMP platform to allow room-temperature storage for deployment in resource limited settings. Our results demonstrated excellent sensitivity and specificity for viral nucleic acid sequences with lower limits of detection of less than 100 copies. Moreover, lyophilized reaction mixtures retained activity for prolonged periods under dry conditions at room temperature. This approach offers a way for detection of emerging viruses in resource limited settings.

  2. Methods And Devices For Characterizing Duplex Nucleic Acid Molecules

    DOEpatents

    Akeson, Mark; Vercoutere, Wenonah; Haussler, David; Winters-Hilt, Stephen

    2005-08-30

    Methods and devices are provided for characterizing a duplex nucleic acid, e.g., a duplex DNA molecule. In the subject methods, a fluid conducting medium that includes a duplex nucleic acid molecule is contacted with a nanopore under the influence of an applied electric field and the resulting changes in current through the nanopore caused by the duplex nucleic acid molecule are monitored. The observed changes in current through the nanopore are then employed as a set of data values to characterize the duplex nucleic acid, where the set of data values may be employed in raw form or manipulated, e.g., into a current blockade profile. Also provided are nanopore devices for practicing the subject methods, where the subject nanopore devices are characterized by the presence of an algorithm which directs a processing means to employ monitored changes in current through a nanopore to characterize a duplex nucleic acid molecule responsible for the current changes. The subject methods and devices find use in a variety of applications, including, among other applications, the identification of an analyte duplex DNA molecule in a sample, the specific base sequence at a single nulceotide polymorphism (SNP), and the sequencing of duplex DNA molecules.

  3. Extract interaction detection methods from the biological literature

    PubMed Central

    Wang, Hongning; Huang, Minlie; Zhu, Xiaoyan

    2009-01-01

    Background Considerable efforts have been made to extract protein-protein interactions from the biological literature, but little work has been done on the extraction of interaction detection methods. It is crucial to annotate the detection methods in the literature, since different detection methods shed different degrees of reliability on the reported interactions. However, the diversity of method mentions in the literature makes the automatic extraction quite challenging. Results In this article, we develop a generative topic model, the Correlated Method-Word model (CMW model) to extract the detection methods from the literature. In the CMW model, we formulate the correlation between the different methods and related words in a probabilistic framework in order to infer the potential methods from the given document. By applying the model on a corpus of 5319 full text documents annotated by the MINT and IntAct databases, we observe promising results, which outperform the best result reported in the BioCreative II challenge evaluation. Conclusion From the promising experiment results, we can see that the CMW model overcomes the issues caused by the diversity in the method mentions and properly captures the in-depth correlations between the detection methods and related words. The performance outperforming the baseline methods confirms that the dependence assumptions of the model are reasonable and the model is competent for the practical processing. PMID:19208158

  4. A Method of Detections' Fusion for GNSS Anti-Spoofing.

    PubMed

    Tao, Huiqi; Li, Hong; Lu, Mingquan

    2016-12-19

    The spoofing attack is one of the security threats of systems depending on the Global Navigation Satellite System (GNSS). There have been many GNSS spoofing detection methods, and each of them focuses on a characteristic of the GNSS signal or a measurement that the receiver has obtained. The method based on a single detector is insufficient against spoofing attacks in some scenarios. How to fuse multiple detections together is a problem that concerns the performance of GNSS anti-spoofing. Scholars have put forward a model to fuse different detection results based on the Dempster-Shafer theory (DST) of evidence combination. However, there are some problems in the application. The main challenge is the valuation of the belief function, which is a key issue in DST. This paper proposes a practical method of detections' fusion based on an approach to assign the belief function for spoofing detections. The frame of discernment is simplified, and the hard decision of hypothesis testing is replaced by the soft decision; then, the belief functions for some detections can be evaluated. The method is discussed in detail, and a performance evaluation is provided, as well. Detections' fusion reduces false alarms of detection and makes the result more reliable. Experimental results based on public test datasets demonstrate the performance of the proposed method.

  5. Development and Validation of a HPTLC Method for Simultaneous Estimation of L-Glutamic Acid and γ-Aminobutyric Acid in Mice Brain.

    PubMed

    Sancheti, J S; Shaikh, M F; Khatwani, P F; Kulkarni, Savita R; Sathaye, Sadhana

    2013-11-01

    A new robust, simple and economic high performance thin layer chromatographic method was developed for simultaneous estimation of L-glutamic acid and γ-amino butyric acid in brain homogenate. The high performance thin layer chromatographic separation of these amino acid was achieved using n-butanol:glacial acetic acid:water (22:3:5 v/v/v) as mobile phase and ninhydrin as a derivatising agent. Quantitation of the method was achieved by densitometric method at 550 nm over the concentration range of 10-100 ng/spot. This method showed good separation of amino acids in the brain homogenate with Rf value of L-glutamic acid and γ-amino butyric acid as 21.67±0.58 and 33.67±0.58, respectively. The limit of detection and limit of quantification for L-glutamic acid was found to be 10 and 20 ng and for γ-amino butyric acid it was 4 and 10 ng, respectively. The method was also validated in terms of accuracy, precision and repeatability. The developed method was found to be precise and accurate with good reproducibility and shows promising applicability for studying pathological status of disease and therapeutic significance of drug treatment.

  6. Method of detecting meter base on image-processing

    NASA Astrophysics Data System (ADS)

    Wang, Hong-ping; Wang, Peng; Yu, Zheng-lin

    2008-03-01

    This paper proposes a new idea of detecting meter using image arithmetic- logic operation and high-precision raster sensor. This method regards the data measured by precision raster as real value, the data obtained by digital image-processing as measuring value, and achieves the aim of detecting meter through the compare of above two datum finally. This method utilizes the dynamic change of meter pointer to complete subtraction processing of image, to realize image segmentation, and to achieve warp-value of image pointer of border time. This method using the technology of image segmentation replaces the traditional method which is low accuracy and low repetition caused by manual operation and ocular reading. Its precision reaches technology index demand according to the arithmetic of nation detecting rules and experiment indicates it is reliable, high accuracy. The paper introduces the total scheme of detecting meter, capturing method of image pointer, and also shows the precision analysis of indicating value error.

  7. A Review of Methods for Detecting Melamine in Food Samples.

    PubMed

    Lu, Yang; Xia, Yinqiang; Liu, Guozhen; Pan, Mingfei; Li, Mengjuan; Lee, Nanju Alice; Wang, Shuo

    2017-01-02

    Melamine is a synthetic chemical used in the manufacture of resins, pigments, and superplasticizers. Human beings can be exposed to melamine through various sources such as migration from related products into foods, pesticide contamination, and illegal addition to foods. Toxicity studies suggest that prolonged consumption of melamine could lead to the formation of kidney stones or even death. Therefore, reliable and accurate detection methods are essential to prevent human exposure to melamine. Sample preparation is of critical importance, since it could directly affect the performance of analytical methods. Some methods for the detection of melamine include instrumental analysis, immunoassays, and sensor methods. In this paper, we have summarized the state-of-the-art methods used for food sample preparation as well as the various detection techniques available for melamine. Combinations of multiple techniques and new materials used in the detection of melamine have also been reviewed. Finally, future perspectives on the applications of microfluidic devices have also been provided.

  8. Sonoclot(®)-based method to detect iron enhanced coagulation.

    PubMed

    Nielsen, Vance G; Henderson, Jon

    2016-07-01

    Thrombelastographic methods have been recently introduced to detect iron mediated hypercoagulability in settings such as sickle cell disease, hemodialysis, mechanical circulatory support, and neuroinflammation. However, these inflammatory situations may have heme oxygenase-derived, coexistent carbon monoxide present, which also enhances coagulation as assessed by the same thrombelastographic variables that are affected by iron. This brief report presents a novel, Sonoclot-based method to detect iron enhanced coagulation that is independent of carbon monoxide influence. Future investigation will be required to assess the sensitivity of this new method to detect iron mediated hypercoagulability in clinical settings compared to results obtained with thrombelastographic techniques.

  9. Systems and methods for detection of blowout precursors in combustors

    DOEpatents

    Lieuwen, Tim C.; Nair, Suraj

    2006-08-15

    The present invention comprises systems and methods for detecting flame blowout precursors in combustors. The blowout precursor detection system comprises a combustor, a pressure measuring device, and blowout precursor detection unit. A combustion controller may also be used to control combustor parameters. The methods of the present invention comprise receiving pressure data measured by an acoustic pressure measuring device, performing one or a combination of spectral analysis, statistical analysis, and wavelet analysis on received pressure data, and determining the existence of a blowout precursor based on such analyses. The spectral analysis, statistical analysis, and wavelet analysis further comprise their respective sub-methods to determine the existence of blowout precursors.

  10. [Detection of amino acids based on terahertz spectroscopy].

    PubMed

    Tang, Zhong-feng; Lin, Hai-tao; Chen, Xiao-wei; Zhang, Zeng-fang

    2009-09-01

    Terahertz (THz) is the frequency region ranging from 0.1 to 2.0 THz, which lies in the far-infrared region. Compared to Fourier transform infrared spectra (FTIR), terahertz time-domain spectra (THz-TDS) has low energy, high signal-to-noise ratio (SNR) and is non-ionizing radiation. Low-frequency vibrational modes of some amino acids, such as torsional and collective vibrational modes and hydrogen-bond modes, exist in the THz region. Amino acids are important organic compounds and are the fundamental components of proteins. Amino acids can exist with a highly ordered crystal structure linked by hydrogen intermolecular bonds in the solid phase. The absorption spectra of amino acids in the THz region show marked differences while mid-infrared absorption spectra usually show very little difference. Up to now, absorption spectra of twenty kinds of amino acids have been studied by many researchers using THz technique; the quantitative analysis of amino acids by THZ-TDS is also included. Investigation of THz spectra of amino acids are of fundamental interests, and will lead to further understanding of low-frequency vibrations of protein/DNA and relevant biological reactions and activities. In the present paper, the latest progress in absorption spectra of amino acids determined by THz spectroscopy is reviewed and a database is built. Some brief remarks on future developments in and prospects for THz application in amino acids are also provided.

  11. Performance evaluation of fault detection methods for wastewater treatment processes.

    PubMed

    Corominas, Lluís; Villez, Kris; Aguado, Daniel; Rieger, Leiv; Rosén, Christian; Vanrolleghem, Peter A

    2011-02-01

    Several methods to detect faults have been developed in various fields, mainly in chemical and process engineering. However, minimal practical guidelines exist for their selection and application. This work presents an index that allows for evaluating monitoring and diagnosis performance of fault detection methods, which takes into account several characteristics, such as false alarms, false acceptance, and undesirable switching from correct detection to non-detection during a fault event. The usefulness of the index to process engineering is demonstrated first by application to a simple example. Then, it is used to compare five univariate fault detection methods (Shewhart, EWMA, and residuals of EWMA) applied to the simulated results of the Benchmark Simulation Model No. 1 long-term (BSM1_LT). The BSM1_LT, provided by the IWA Task Group on Benchmarking of Control Strategies, is a simulation platform that allows for creating sensor and actuator faults and process disturbances in a wastewater treatment plant. The results from the method comparison using BSM1_LT show better performance to detect a sensor measurement shift for adaptive methods (residuals of EWMA) and when monitoring the actuator signals in a control loop (e.g., airflow). Overall, the proposed index is able to screen fault detection methods.

  12. Optical methods for the detection of heavy metal ions

    NASA Astrophysics Data System (ADS)

    Uglov, A. N.; Bessmertnykh-Lemeune, A.; Guilard, R.; Averin, A. D.; Beletskaya, I. P.

    2014-03-01

    The review covers an important area of the modern chemistry, namely, the detection of heavy metal ions using optical molecular detectors. The role of this method in metal ion detection and the physicochemical grounds of operation of chemosensors are discussed, and examples of detection of most abundant heavy metal ions and synthetic approaches to molecular detectors are presented. The immobilization of molecular detectors on solid substrates for the design of analytical sensor devices is described. The bibliography includes 178 references.

  13. Feature matching method in shaped light mode VFD defect detection

    NASA Astrophysics Data System (ADS)

    Jin, Xuanhong; Dai, Shuguang; Mu, Pingan

    2010-08-01

    In recent years, Vacuum Fluorescent Display (VFD) module in the car audio panel has been widely used. However, due to process reasons, VFD display production process will produce defects, not only affect the appearance, but also affect the display correctly. So building a car VFD display panel defect detection system is of great significance. Machine vision technology is introduced into the automotive VFD display defect detection in order to achieve fast and accurate detection of defects. Shaped light mode is a typical flaw detection mode which is based on characteristics of vehicle VFD panel. According to the image features, learning of the gray matching and feature matching method, we integrated use of feature matching method and the gray level matching method to achieve defect detection.

  14. A method for detection and diagnosis on batch fermentations.

    PubMed

    Dondo, Rodolfo G

    2003-01-01

    In this work we present some basic ideas about detection and diagnosis of faults and abrupt dynamic changes in batch fermentations. Our work focuses on the simultaneous use of two detection methods (residual based and balances based) within the estimation procedure. The idea behind the use of both methods is that the weakness of one of them can be compensated by the use of the other one. Thus the simultaneous use of both methods allows detecting and possibly isolating a wide range of faults. Observations such as the effect of nonlinearities on the detection tests and robustness to model uncertainty are discussed. Numerical results on a particular case, the xanthan gum batch fermentation, are presented. Simulated faults and abnormal behaviors were promptly detected but diagnostics showed mixed results.

  15. Microwave-Accelerated Method for Ultra-Rapid Extraction of Neisseria gonorrhoeae DNA for Downstream Detection

    PubMed Central

    Melendez, Johan H.; Santaus, Tonya M.; Brinsley, Gregory; Kiang, Daniel; Mali, Buddha; Hardick, Justin; Gaydos, Charlotte A.; Geddes, Chris D.

    2016-01-01

    Nucleic acid-based detection of gonorrhea infections typically require a two-step process involving isolation of the nucleic acid, followed by the detection of the genomic target often involving PCR-based approaches. In an effort to improve on current detection approaches, we have developed a unique two-step microwave-accelerated approach for rapid extraction and detection of Neisseria gonorrhoeae (GC) DNA. Our approach is based on the use of highly-focused microwave radiation to rapidly lyse bacterial cells, release, and subsequently fragment microbial DNA. The DNA target is then detected by a process known as microwave-accelerated metal-enhanced fluorescence (MAMEF), an ultra-sensitive direct DNA detection analytical technique. In the present study, we show that highly focused microwaves at 2.45 GHz, using 12.3 mm gold film equilateral triangles, are able to rapidly lyse both bacteria cells and fragment DNA in a time- and microwave power-dependent manner. Detection of the extracted DNA can be performed by MAMEF, without the need for DNA amplification in less than 10 minutes total time or by other PCR-based approaches. Collectively, the use of a microwave-accelerated method for the release and detection of DNA represents a significant step forward towards the development of a point-of-care (POC) platform for detection of gonorrhea infections. PMID:27325503

  16. Microwave-accelerated method for ultra-rapid extraction of Neisseria gonorrhoeae DNA for downstream detection.

    PubMed

    Melendez, Johan H; Santaus, Tonya M; Brinsley, Gregory; Kiang, Daniel; Mali, Buddha; Hardick, Justin; Gaydos, Charlotte A; Geddes, Chris D

    2016-10-01

    Nucleic acid-based detection of gonorrhea infections typically require a two-step process involving isolation of the nucleic acid, followed by detection of the genomic target often involving polymerase chain reaction (PCR)-based approaches. In an effort to improve on current detection approaches, we have developed a unique two-step microwave-accelerated approach for rapid extraction and detection of Neisseria gonorrhoeae (gonorrhea, GC) DNA. Our approach is based on the use of highly focused microwave radiation to rapidly lyse bacterial cells, release, and subsequently fragment microbial DNA. The DNA target is then detected by a process known as microwave-accelerated metal-enhanced fluorescence (MAMEF), an ultra-sensitive direct DNA detection analytical technique. In the current study, we show that highly focused microwaves at 2.45 GHz, using 12.3-mm gold film equilateral triangles, are able to rapidly lyse both bacteria cells and fragment DNA in a time- and microwave power-dependent manner. Detection of the extracted DNA can be performed by MAMEF, without the need for DNA amplification, in less than 10 min total time or by other PCR-based approaches. Collectively, the use of a microwave-accelerated method for the release and detection of DNA represents a significant step forward toward the development of a point-of-care (POC) platform for detection of gonorrhea infections.

  17. Colorimetric detection of Cr3+ using gold nanoparticles functionalized with 4-amino hippuric acid

    NASA Astrophysics Data System (ADS)

    Jin, Weiwei; Huang, Pengcheng; Chen, Yueji; Wu, Fangying; Wan, Yiqun

    2015-09-01

    A facile and effective technique for monitoring Cr3+ concentration based on 4-amino hippuric acid (PAH) decorated Au nanoparticles (PAH-AuNPs) is introduced. The modified AuNPs were easily aggregated in the presence of Cr3+, resulting in the color change from red to violet or blue, which is in response to the surface plasmon absorption of dispersed or aggregated nanoparticles. Under the optimized conditions, a good linear relationship (correlation coefficient r = 0.998) was obtained between the ratio of the absorbance at 635 nm to that at 520 nm ( A 635 nm/ A 520 nm), and the concentration of Cr3+ was over the range of 5.0-120 µM with detection limit of 1.17 µM. This method exhibited excellent selectivity for Cr3+ over other tested heavy metal ions. Furthermore, there was no significant difference for the parameters of calibration equation between the presence and absence of ethylenediamine tetraacetic acid (EDTA), which suggests that the method can be applied in various real samples owing to the strong masking ability of EDTA. The assay was used to detect the concentrations of Cr3+ in liquid milk, milk power, and lake water samples with recoveries ranging from 93.5 to 114 %, indicating that the method could be used for extensive practical application.

  18. High-performance liquid chromatography with conductimetric detection of perfluorocarboxylic acids and perfluorosulfonates.

    PubMed

    Hori, Hisao; Hayakawa, Etsuko; Yamashita, Nobuyoshi; Taniyasu, Sachi; Nakata, Fumiya; Kobayashi, Yoshimi

    2004-10-01

    A rapid and simple method for separating and determining various environmentally harmful perfluorocarboxylic acids and perfluorosulfonates was successfully developed using high- performance liquid chromatography with conductimetric detection, for product and waste management of these compounds at manufacturing and processing sites. Compounds having C(3)-C(8) perfluoroalkyl groups were separated using a Tosoh TSKgel Super-ODS column and a mobile phase consisting of a mixture of methanol and aqueous NaH(2)PO(4) at several mixing ratios. The best detection limits for the compounds ranged from 0.12 to 0.66 mg l(-1) (ppm), and linear calibration graphs were obtained up to 87-109 mg l(-1). The combination of this method with concentration of the sample by solid-phase extraction with cartridges based on styrene-divinylbenzene-copolymer enabled the determination of approximately 50 microg l(-1) (ppb) for compounds with C(4)-C(8) perfluoroalkyl groups. This method was successfully used to monitor the artificial decomposition of the perfluorocarboxylic acid n-C(4)F(9)COOH induced by a photocatalyst.

  19. Study on the spectrophotometric detection of free fatty acids in palm oil utilizing enzymatic reactions.

    PubMed

    Azeman, Nur Hidayah; Yusof, Nor Azah; Abdullah, Jaafar; Yunus, Robiah; Hamidon, Mohd Nizar; Hajian, Reza

    2015-07-07

    In this paper, a comprehensive study has been made on the detection of free fatty acids (FFAs) in palm oil via an optical technique based on enzymatic aminolysis reactions. FFAs in crude palm oil (CPO) were converted into fatty hydroxamic acids (FHAs) in a biphasic lipid/aqueous medium in the presence of immobilized lipase. The colored compound formed after complexation between FHA and vanadium (V) ion solution was proportional to the FFA content in the CPO samples and was analyzed using a spectrophotometric method. In order to develop a rapid detection system, the parameters involved in the aminolysis process were studied. The utilization of immobilized lipase as catalyst during the aminolysis process offers simplicity in the product isolation and the possibility of conducting the process under extreme reaction conditions. A good agreement was found between the developed method using immobilized Thermomyces lanuginose lipase as catalyst for the aminolysis process and the Malaysian Palm Oil Board (MPOB) standard titration method (R2 = 0.9453).

  20. Method of enhancing radiation response of radiation detection materials

    DOEpatents

    Miller, Steven D.

    1997-01-01

    The present invention is a method of increasing radiation response of a radiation detection material for a given radiation signal by first pressurizing the radiation detection material. Pressurization may be accomplished by any means including mechanical and/or hydraulic. In this application, the term "pressure" includes fluid pressure and/or mechanical stress.

  1. Method for the detection of Salmonella enterica serovar Enteritidis

    DOEpatents

    Agron, Peter G.; Andersen, Gary L.; Walker, Richard L.

    2008-10-28

    Described herein is the identification of a novel Salmonella enterica serovar Enteritidis locus that serves as a marker for DNA-based identification of this bacterium. In addition, three primer pairs derived from this locus that may be used in a nucleotide detection method to detect the presence of the bacterium are also disclosed herein.

  2. Comparative assessment of the methods for exchangeable acidity measuring

    NASA Astrophysics Data System (ADS)

    Vanchikova, E. V.; Shamrikova, E. V.; Bespyatykh, N. V.; Zaboeva, G. A.; Bobrova, Yu. I.; Kyz"yurova, E. V.; Grishchenko, N. V.

    2016-05-01

    A comparative assessment of the results of measuring the exchangeable acidity and its components by different methods was performed for the main mineral genetic horizons of texturally-differentiated gleyed and nongleyed soddy-podzolic and gley-podzolic soils of the Komi Republic. It was shown that the contents of all the components of exchangeable soil acidity determined by the Russian method (with potassium chloride solution as extractant, c(KCl) = 1 mol/dm3) were significantly higher than those obtained by the international method (with barium chloride solution as extractant, c(BaCl2) = 0.1 mol/dm3). The error of the estimate of the concentration of H+ ions extracted with barium chloride solution equaled 100%, and this allowed only qualitative description of this component of the soil acidity. In the case of the extraction with potassium chloride, the error of measurements was 50%. It was also shown that the use of potentiometric titration suggested by the Russian method overestimates the results of soil acidity measurement caused by the exchangeable metal ions (Al(III), Fe(III), and Mn(II)) in comparison with the atomic emission method.

  3. Preparation of κ-carra-oligosaccharides with microwave assisted acid hydrolysis method

    NASA Astrophysics Data System (ADS)

    Li, Guangsheng; Zhao, Xia; Lv, Youjing; Li, Miaomiao; Yu, Guangli

    2015-04-01

    A rapid method of microwave assisted acid hydrolysis was established to prepare κ-carra-oligosaccharides. The optimal hydrolysis condition was determined by an orthogonal test. The degree of polymerization (DP) of oligosaccharides was detected by high performance thin layer chromatography (HPTLC) and polyacrylamide gel electrophoresis (PAGE). Considering the results of HPTLC and PAGE, the optimum condition of microwave assisted acid hydrolysis was determined. The concentration of κ-carrageenan was 5 mg mL-1; the reaction solution was adjusted to pH 3 with diluted hydrochloric acid; the solution was hydrolyzed under microwave irradiation at 100 for 15 °C min. Oligosaccharides were separated by a Superdex 30 column (2.6 cm × 90 cm) using AKTA Purifier UPC100 and detected with an online refractive index detector. Each fraction was characterized by electrospray ionization mass spectrometry (ESI-MS). The data showed that odd-numbered κ-carra-oligosaccharides with DP ranging from 3 to 21 could be obtained with this method, and the structures of the oligosaccharides were consistent with those obtained by traditional mild acid hydrolysis. The new method was more convenient, efficient and environment-friendly than traditional mild acid hydrolysis. Our results provided a useful reference for the preparation of oligosaccharides from other polysaccharides.

  4. Nucleic acid based fluorescent sensor for mercury detection

    DOEpatents

    Lu, Yi; Liu, Juewen

    2013-02-05

    A nucleic acid enzyme comprises an oligonucleotide containing thymine bases. The nucleic acid enzyme is dependent on both Hg.sup.2+and a second ion as cofactors, to produce a product from a substrate. The substrate comprises a ribonucleotide, a deoxyribonucleotide, or both.

  5. Radionuclide detection by inductively coupled plasma mass spectrometry: A comparison of atomic and radiation detection method

    SciTech Connect

    Smith, M.R.; Wyse, E.J.; Koppenaal, D.W.

    1991-04-01

    Radionuclide detection by mass spectrometric techniques offers inherent advantages over conventional radiation detection methods. Since radionuclides decay at variable rates (half-lives) and via various nuclear transformations (i.e. emission of alpha, beta, and/or gamma radiation) their determination via radiation detection depends not only on decay systematics but also on detector technology. Radionuclide detection by direct atom measurement, however, is dependent only on technique sensitivity and is indifferent to decay mode. Evaluation of inductively coupled plasma mass spectrometry (ICP/MS) indicates this method to be superior conventional radiation detection techniques for many radionuclides. This work discusses factors which influence detection by both methods. Illustrative applications of ICP/MS to the ultra-trace determination of several radionuclides, including {sup 129}I, are presented. 20 refs., 6 figs., 1 tab.

  6. Highly sensitive SERS detection and quantification of sialic acid on single cell using photonic-crystal fiber with gold nanoparticles.

    PubMed

    Gong, Tianxun; Cui, Ying; Goh, Douglas; Voon, Kong Kien; Shum, Perry Ping; Humbert, Georges; Auguste, Jean-Louis; Dinh, Xuan-Quyen; Yong, Ken-Tye; Olivo, Malini

    2015-02-15

    An ultrasensitive surface enhanced Raman spectroscopy (SERS) based sensing platform was developed to detect the mean sialic acid level on the surface of single cell with sensitivity as low as 2 fmol. This platform adopted the use of an interference-free Raman tag, 4-(dihydroxyborophenyl) acetylene (DBA), which selectively binds to sialic acid on the cell membrane. By loading the side channel of a photonic crystal fiber with a mixture of gold nanoparticles and DBA-tagged HeLa cell, and subsequently propagating laser light through the central solid core, strong SERS signal was obtained. This SERS technique achieved accurate detection and quantification of concentration of sialic acid on a single cell, surpassing previously reported methods that required more than 10(5) cells. Moreover, this platform can be developed into a clinical diagnostic tool to potentially analyze sialic acid-related diseases such as tumor malignancy and metastasis in real-time.

  7. A fast automatic target detection method for detecting ships in infrared scenes

    NASA Astrophysics Data System (ADS)

    Özertem, Kemal Arda

    2016-05-01

    Automatic target detection in infrared scenes is a vital task for many application areas like defense, security and border surveillance. For anti-ship missiles, having a fast and robust ship detection algorithm is crucial for overall system performance. In this paper, a straight-forward yet effective ship detection method for infrared scenes is introduced. First, morphological grayscale reconstruction is applied to the input image, followed by an automatic thresholding onto the suppressed image. For the segmentation step, connected component analysis is employed to obtain target candidate regions. At this point, it can be realized that the detection is defenseless to outliers like small objects with relatively high intensity values or the clouds. To deal with this drawback, a post-processing stage is introduced. For the post-processing stage, two different methods are used. First, noisy detection results are rejected with respect to target size. Second, the waterline is detected by using Hough transform and the detection results that are located above the waterline with a small margin are rejected. After post-processing stage, there are still undesired holes remaining, which cause to detect one object as multi objects or not to detect an object as a whole. To improve the detection performance, another automatic thresholding is implemented only to target candidate regions. Finally, two detection results are fused and post-processing stage is repeated to obtain final detection result. The performance of overall methodology is tested with real world infrared test data.

  8. Nested methylation-specific polymerase chain reaction cancer detection method

    DOEpatents

    Belinsky, Steven A.; Palmisano, William A.

    2007-05-08

    A molecular marker-based method for monitoring and detecting cancer in humans. Aberrant methylation of gene promoters is a marker for cancer risk in humans. A two-stage, or "nested" polymerase chain reaction method is disclosed for detecting methylated DNA sequences at sufficiently high levels of sensitivity to permit cancer screening in biological fluid samples, such as sputum, obtained non-invasively. The method is for detecting the aberrant methylation of the p16 gene, O 6-methylguanine-DNA methyltransferase gene, Death-associated protein kinase gene, RAS-associated family 1 gene, or other gene promoters. The method offers a potentially powerful approach to population-based screening for the detection of lung and other cancers.

  9. [Fast Implementation Method of Protein Spots Detection Based on CUDA].

    PubMed

    Xiong, Bangshu; Ye, Yijia; Ou, Qiaofeng; Zhang, Haodong

    2016-02-01

    In order to improve the efficiency of protein spots detection, a fast detection method based on CUDA was proposed. Firstly, the parallel algorithms of the three most time-consuming parts in the protein spots detection algorithm: image preprocessing, coarse protein point detection and overlapping point segmentation were studied. Then, according to single instruction multiple threads executive model of CUDA to adopted data space strategy of separating two-dimensional (2D) images into blocks, various optimizing measures such as shared memory and 2D texture memory are adopted in this study. The results show that the operative efficiency of this method is obviously improved compared to CPU calculation. As the image size increased, this method makes more improvement in efficiency, such as for the image with the size of 2,048 x 2,048, the method of CPU needs 52,641 ms, but the GPU needs only 4,384 ms.

  10. A novel method to detect shadows on multispectral images

    NASA Astrophysics Data System (ADS)

    Daǧlayan Sevim, Hazan; Yardımcı ćetin, Yasemin; Özışık Başkurt, Didem

    2016-10-01

    Shadowing occurs when the direct light coming from a light source is obstructed by high human made structures, mountains or clouds. Since shadow regions are illuminated only by scattered light, true spectral properties of the objects are not observed in such regions. Therefore, many object classification and change detection problems utilize shadow detection as a preprocessing step. Besides, shadows are useful for obtaining 3D information of the objects such as estimating the height of buildings. With pervasiveness of remote sensing images, shadow detection is ever more important. This study aims to develop a shadow detection method on multispectral images based on the transformation of C1C2C3 space and contribution of NIR bands. The proposed method is tested on Worldview-2 images covering Ankara, Turkey at different times. The new index is used on these 8-band multispectral images with two NIR bands. The method is compared with methods in the literature.

  11. An improved PCA method with application to boiler leak detection.

    PubMed

    Sun, Xi; Marquez, Horacio J; Chen, Tongwen; Riaz, Muhammad

    2005-07-01

    Principal component analysis (PCA) is a popular fault detection technique. It has been widely used in process industries, especially in the chemical industry. In industrial applications, achieving a sensitive system capable of detecting incipient faults, which maintains the false alarm rate to a minimum, is a crucial issue. Although a lot of research has been focused on these issues for PCA-based fault detection and diagnosis methods, sensitivity of the fault detection scheme versus false alarm rate continues to be an important issue. In this paper, an improved PCA method is proposed to address this problem. In this method, a new data preprocessing scheme and a new fault detection scheme designed for Hotelling's T2 as well as the squared prediction error are developed. A dynamic PCA model is also developed for boiler leak detection. This new method is applied to boiler water/steam leak detection with real data from Syncrude Canada's utility plant in Fort McMurray, Canada. Our results demonstrate that the proposed method can effectively reduce false alarm rate, provide effective and correct leak alarms, and give early warning to operators.

  12. A new ultrasound based method for rapid microorganism detection

    NASA Astrophysics Data System (ADS)

    Shukla, Shiva Kant; Segura, Luis Elvira; Sánchez, Carlos José Sierra; López, Pablo Resa

    2012-05-01

    A new method for rapid detection of catalase positive microorganisms by using an ultrasonic measuring method is proposed in this work. The developed technique is based on the detection of oxygen bubbles produced by the hydrolysis of hydrogen peroxide induced by the enzyme catalase which is present in many microorganisms. The bubbles are trapped in a media based on agar gel which was especially developed for microbiological evaluation. It is found that microorganism concentrations of the order of 105 c.f.u./ml can be detected by using this method. The results obtained show up that the proposed method is competitive with other modern commercial methods like luminescence by ATP system. The method can also be used for characterization of enzyme activity.

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

    PubMed

    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.

  14. A novel eyelash detection method for iris recognition.

    PubMed

    Yuan, Weiqi; He, Wei

    2005-01-01

    Iris is often affected by the eyelash noise, when captured under unfavorable condition. Not only the iris localization of inner and outer boundaries, but also iris feature extraction can be affected by eyelash. Therefore, in an iris recognition system, eyelash detection is of great importance for accurate iris recognition. Eyelash can be classified into two classes: (1) separable eyelashes (2) multiple eyelashes. The former are detected by local intensity minimum algorithm, and the latter are detected according to the template mean and standard deviation. Experimental results show that the proposed method can detect eyelash accurately.

  15. Cationic poly(lactic-co-glycolic acid) iron oxide microspheres for nucleic acid detection

    NASA Astrophysics Data System (ADS)

    Pandey, Chandra Mouli; Sharma, Aditya; Sumana, Gajjala; Tiwari, Ida; Malhotra, Bansi Dhar

    2013-04-01

    Herein, we envisage the possibility of preparing stable cationic poly(lactic-co-glycolic acid) (PLGA) microspheres encapsulating the iron oxide nanoparticles (IONPs; 8-12 nm). The IONPs are incorporated into PLGA in organic phase followed by microsphere formation and chitosan coating in aqueous medium via nano-emulsion technique. The average size of the microspheres, as determined by dynamic light scattering are about 310 nm, while the zeta potential for the composite remains near 35 mV at pH 4.0. These microspheres are electrophoretically deposited onto indium tin oxide (ITO)-coated glass substrate used as cathode and parallel platinum plate as the counter electrode. This platform is utilized to fabricate a DNA biosensor, by immobilizing a probe sequence specific to Escherichia coli. The bioelectrode shows a surface-controlled electrode reaction with the electron transfer coefficient (α) of 0.64 and charge transfer rate constant (ks) of 61.73 s-1. Under the optimal conditions, this biosensor shows a detection limit of 8.7 × 10-14 M and is found to retain about 81% of the initial activity after 9 cycles of use.Herein, we envisage the possibility of preparing stable cationic poly(lactic-co-glycolic acid) (PLGA) microspheres encapsulating the iron oxide nanoparticles (IONPs; 8-12 nm). The IONPs are incorporated into PLGA in organic phase followed by microsphere formation and chitosan coating in aqueous medium via nano-emulsion technique. The average size of the microspheres, as determined by dynamic light scattering are about 310 nm, while the zeta potential for the composite remains near 35 mV at pH 4.0. These microspheres are electrophoretically deposited onto indium tin oxide (ITO)-coated glass substrate used as cathode and parallel platinum plate as the counter electrode. This platform is utilized to fabricate a DNA biosensor, by immobilizing a probe sequence specific to Escherichia coli. The bioelectrode shows a surface-controlled electrode reaction with the

  16. Seed-mediated grown silver nanoparticles as a colorimetric sensor for detection of ascorbic acid.

    PubMed

    Rostami, Simindokht; Mehdinia, Ali; Jabbari, Ali

    2017-06-05

    A simple and sensitive approach was demonstrated for detection of ascorbic acid (AA) based on seed-mediated growth of silver nanoparticles (Ag NPs). According to the seeding strategy, silver ions existing in the growth solution were reduced to silver atoms on the surface of silver seeds via redox reaction between silver ions and AA. This process -led to appear an absorption band in near 420nm owing to the localized surface plasmon resonance peak of the generated Ag NPs. This change in absorption spectra of Ag NPs caused a change in color of the mixture from colorless to yellow. It was found that the changes in absorption intensity at 420nm have a good relationship with the concentration of AA. Also, detection of AA was achieved through the established colorimetric sensor in the range of 0.25-25μM with detection limit of 0.054μM. Moreover, the selectivity of the method was evaluated with considering potential interferences. The method showed high selectivity toward AA rather than potential interferences and coexisted molecules with AA. It was successfully applied for detection and determination of AA in pharmaceutical tablets and commercial lemonade.

  17. Method for preparing 6-.beta.-halopenicillanic acids

    DOEpatents

    Hansen, Erik I.; Kran-Nielsen, Mogens P.; Von Daehne, Welf

    1989-01-01

    The present invention relates to a new and improved method for the preparation of a compound of the formula I ##STR1## in which R stands for halogen, giving rise to high yields of substantially pure 6.beta.-halopenicillanic acids, obtained in one step.

  18. Food adulteration: Sources, health risks, and detection methods.

    PubMed

    Bansal, Sangita; Singh, Apoorva; Mangal, Manisha; Mangal, Anupam K; Kumar, Sanjiv

    2017-04-13

    Adulteration in food has been a concern since the beginning of civilization, as it not only decreases the quality of food products but also results in a number of ill effects on health. Authentic testing of food and adulterant detection of various food products is required for value assessment and to assure consumer protection against fraudulent activities. Through this review we intend to compile different types of adulterations made in different food items, the health risks imposed by these adulterants and detection methods available for them. Concerns about food safety and regulation have ensured the development of various techniques like physical, biochemical/immunological and molecular techniques, for adulterant detection in food. Molecular methods are more preferable when it comes to detection of biological adulterants in food, although physical and biochemical techniques are preferable for detection of other adulterants in food.

  19. Automatic landslide and mudflow detection method via multichannel sparse representation

    NASA Astrophysics Data System (ADS)

    Chao, Chen; Zhou, Jianjun; Hao, Zhuo; Sun, Bo; He, Jun; Ge, Fengxiang

    2015-10-01

    Landslide and mudflow detection is an important application of aerial images and high resolution remote sensing images, which is crucial for national security and disaster relief. Since the high resolution images are often large in size, it's necessary to develop an efficient algorithm for landslide and mudflow detection. Based on the theory of sparse representation and, we propose a novel automatic landslide and mudflow detection method in this paper, which combines multi-channel sparse representation and eight neighbor judgment methods. The whole process of the detection is totally automatic. We make the experiment on a high resolution image of ZhouQu district of Gansu province in China on August, 2010 and get a promising result which proved the effective of using sparse representation on landslide and mudflow detection.

  20. An analytical method for the measurement of acid metabolites of tryptophan-NAD pathway and related acids in urine.

    PubMed

    Liao, Xiangjun; Zhu, Jiping; Rubab, Mamoona; Feng, Yong-Lai; Poon, Raymond

    2010-04-15

    An analytical method has been developed for the measurements of five urinary acids namely, quinolinic acid, picolinic acid, nicotinic acid, 2-pyridylacetic acid and 3-pyridylacetic acid. The high performance liquid chromatograph-electrospray ionization mass spectrometry was operated in positive polarity under selected ion monitoring mode, with a column flow rate of 0.2 ml/min and an injection volume of 20 microl. The method used isotope-labelled picolinic acid (PA-d(4)) and nicotinic acid (NA-d(4)) as internal standards for the quantification. The sample preparation involved parallel use of two different types of mixed-mode solid phase extraction cartridges (Strata-X-AW for the extraction of quinolinic acid, and Strata-X-C for the remaining acids). Quantitative analysis of five target acids in several human and rat urine samples showed that the levels of acids were relatively uniform among rats while larger variations were observed for human samples.

  1. A simplified method for estimation of jarosite and acid-forming sulfates in acid mine wastes.

    PubMed

    Li, Jun; Smart, Roger St C; Schumann, Russell C; Gerson, Andrea R; Levay, George

    2007-02-01

    In acid base accounting (ABA) estimates of acid mine wastes, the acid potential (AP) estimate can be improved by using the net carbonate value (NCV) reactive sulfide S method rather than total S assay methods but this does not give recovery of potentially acid producing ferrous and ferric sulfates present in many wastes. For more accurate estimation of AP, an effective, site-specific method to quantify acid sulfate salts, such as jarosite and melanterite, in waste rocks has been developed and tested on synthetic and real wastes. The SPOCAS (acid sulfate soils) methods have been modified to an effective, rapid method to speciate sulfate forms in different synthetic waste samples. A three-step sequential extraction procedure has been established. These steps are: (1) argon-purged water extraction (3 min) to extract soluble Fe(II) salts (particularly melanterite), epsomite and gypsum (<10 wt.%), (2) roasting at 550 degrees C (1 h) to remove sulfur from pyrite and other reactive sulfides, (3) HCl extraction (4 M, 30 min) for determination of jarosites. Products (solid and aqueous) have been characterized at each step including the jarosite decomposition process in Step 2 where temperature control is critical to avoid S loss. The sequential extraction procedure was used to quantitatively determine melanterite, epsomite, gypsum, pyrite and jarosite concentrations in a synthetic waste sample containing these mineral phases at 5 wt.% in quartz, and also tested using a tailings waste sample to quantitatively determine epsomite, gypsum and jarosite contents. The method is applicable to most waste samples including those with non-pyrite sulfides but for samples containing significant amounts of sulfur (>1 wt.% S) as copper sulfides, the second step of roasting needs to be excluded from the procedure with an increased time of 4 M HCl extraction to 16 h for jarosite determination.

  2. Capillary electrophoresis with laser-induced fluorescence detection for studying amino acid uptake by yeast during beer fermentation.

    PubMed

    Turkia, Heidi; Sirén, Heli; Penttilä, Merja; Pitkänen, Juha-Pekka

    2015-01-01

    The amino acid composition of cultivation broth is known to affect the biomass accumulation, productivity, and vitality of yeast during cultivation. A separation method based on capillary electrophoresis with laser-induced fluorescence (LIF) detection was developed for the determination of amino acid consumption by Saccharomyces cerevisiae during beer fermentation. Intraday relative standard deviations were less than 2.1% for migration times and between 2.9% and 9.9% for peak areas. Interday relative standard deviations were less than 2.5% for migration times and between 4.4% and 18.9% for peak areas. The quantification limit was even as low as 62.5 pM which equals to below attomole level detection. The method was applied to study the rate of amino acid utilization during beer fermentation.

  3. Efficient method of image edge detection based on FSVM

    NASA Astrophysics Data System (ADS)

    Cai, Aiping; Xiong, Xiaomei

    2013-07-01

    For efficient object cover edge detection in digital images, this paper studied traditional methods and algorithm based on SVM. It analyzed Canny edge detection algorithm existed some pseudo-edge and poor anti-noise capability. In order to provide a reliable edge extraction method, propose a new detection algorithm based on FSVM. Which contains several steps: first, trains classify sample and gives the different membership function to different samples. Then, a new training sample is formed by increase the punishment some wrong sub-sample, and use the new FSVM classification model for train and test them. Finally the edges are extracted of the object image by using the model. Experimental result shows that good edge detection image will be obtained and adding noise experiments results show that this method has good anti-noise.

  4. Methods, systems and devices for detecting and locating ferromagnetic objects

    DOEpatents

    Roybal, Lyle Gene [Idaho Falls, ID; Kotter, Dale Kent [Shelley, ID; Rohrbaugh, David Thomas [Idaho Falls, ID; Spencer, David Frazer [Idaho Falls, ID

    2010-01-26

    Methods for detecting and locating ferromagnetic objects in a security screening system. One method includes a step of acquiring magnetic data that includes magnetic field gradients detected during a period of time. Another step includes representing the magnetic data as a function of the period of time. Another step includes converting the magnetic data to being represented as a function of frequency. Another method includes a step of sensing a magnetic field for a period of time. Another step includes detecting a gradient within the magnetic field during the period of time. Another step includes identifying a peak value of the gradient detected during the period of time. Another step includes identifying a portion of time within the period of time that represents when the peak value occurs. Another step includes configuring the portion of time over the period of time to represent a ratio.

  5. Development of an HPLC-fluorescence determination method for carboxylic acids related to the tricarboxylic acid cycle as a metabolome tool.

    PubMed

    Kubota, Kazuyuki; Fukushima, Takeshi; Yuji, Reiko; Miyano, Hiroshi; Hirayama, Kazuo; Santa, Tomofumi; Imai, Kazuhiro

    2005-12-01

    We report the simultaneous determination of the carboxylic acids related to the tricarboxylic acid (TCA) cycle, which plays an important role in producing adenosine triphosphate (ATP) and generating energy in mitochondria. Seven carboxylic acids from the TCA cycle, and pyruvic acid and 2-methylsuccinic acid, as an internal standard, were derivatized with a fluorescent reagent for carboxyl groups, 4-N,N-dimethylaminosulfonyl-7-piperazino-2,1,3-benzoxadiazole (DBD-PZ), in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 4-N,N-dimethyaminopyridine as the coupling reagents, at 60 degrees C for 120 min. Subsequently, the excess DBD-PZ was removed efficiently using a cation-exchange cartridge, SDB-RPS (Empore). These fluorescent derivatives were separated well from each other on an octadecyl silica column (TSKgel ODS-80Ts, 250 x 4.6 mm, i.d.) with an eluent of acetonitrile-water containing 1% formic acid at a flow rate of 0.8 mL/min, and were detected fluorometrically at 560 nm, with excitation at 450 nm. The validation data were satisfactory in the range of 2.5-100 microm citric acid, isocitric acid, 2-oxoglutaric acid, succinic acid and fumaric acid. The detection limit (S/N = 3) for citric acid was 2 fmol on the column. The structures of these derivatives were confirmed by high-performance liquid chromatography-mass spectrometry, which proved that their carboxylic groups were completely labeled with DBD-PZ, except for oxaloacetic acid. This HPLC method was successfully applied to the analysis of TCA cycle metabolites in rat urine. The method will also be useful for metabolome research, such as for target analyses of metabolites with carboxyl groups, not only in urine but also in cells and organs.

  6. Methods for detecting residues of cleaning agents during cleaning validation.

    PubMed

    Westman, L; Karlsson, G

    2000-01-01

    Cleaning validation procedures are carried out in order to assure that residues of cleaning agents are within acceptable limits after the cleaning process. Cleaning agents often consist of a mixture of various surfactants which are in a highly diluted state after the water rinsing procedure has been completed. This makes it difficult to find appropriate analytical methods that are sensitive enough to detect the cleaning agents. In addition, it is advantageous for the analytical methods to be simple to perform and to give results quickly. In this study, four different analytical methods are compared: visual detection of foam, pH, conductivity measurements, and analysis of total organic carbon (TOC). TOC was used as a reference method when evaluating the other three potential methods. The analyses were performed on different dilutions of the cleaning agents Vips Neutral, RBS-25, Debisan and Perform. The results demonstrated that the most sensitive method for analysis of Vips Neutral, Debisan and Perform is visual detection of foam, by which it is possible to detect concentrations of cleaning agents down to 10 micrograms/mL. RBS-25 was not detected below 200 micrograms/mL, probably because it is formulated with low-foaming surfactants. TOC analysis is less sensitive but has the advantage of being a quantitative analysis, while visual detection of foam is a semi-quantitative method. Visual detection of foam is easy to perform, gives a quick result, and requires no expensive instrumentation. The sensitivity of each method was found to be dependent upon the type of cleaning agent that was analyzed.

  7. Determination of component concentrations in mixtures of weak and strong acids and bases by linear algebraic methods.

    PubMed

    Ivaska, A; Nagypál, I

    1980-09-01

    A general expression for transforming potentiometric titration curves of mixtures of weak acids into a system of linear equations is derived. The solution of the linear equations gives directly the concentrations of the components. This linear transformation method is illustrated by the analysis of mixtures of weak acids with overlapping dissociation equilibria. The possible presence of a strong acid or strong base in the mixture can also be detected and its concentration simultaneously determined. The method can also be used for analysis of an ampholyte and solutions containing a weak acid and its conjugate base. For example a mixture of hydroxyacetic acid (pK approximately 3.6), acetic acid (pK approximately 4.6) and hydroxylamine hydrochloride (pK approximately 6) was analysed in the presence of strong acid with an average relative error of approximately 2%.

  8. Comparison between high-performance liquid chromatography and gas chromatography methods for fatty acid identification and quantification in potato crisps.

    PubMed

    Sanches-Silva, A; Rodríguez-Bernaldo de Quirós, A; López-Hernández, J; Paseiro-Losada, P

    2004-04-02

    A reversed-phase high performance liquid chromatographic (RP-HLPC) method was compared with a gas chromatography-flame ionization detection (GC-FID) method for determining fatty acids in potato crisps. Different extraction procedures were used. Fatty acids were quantified by linear regression. Both methods presented good precision (R.S.D. < or = 5.88%) and recovery (> or = 82.31%). The precision using HPLC method was slightly better than for GC-FID method. There was good agreement between the fatty acid composition of potato crisps analysed by both methods. For most purposes the HPLC method would be better. However, when more fatty acids need to be analysed, GC is a more suitable method.

  9. Estimation of organophosphoric acid triesters in soft polyurethane foam using a concentrated sulfuric acid dissolution technique and gas chromatography with flame photometric detection.

    PubMed

    Nagase, Makoto; Toba, Mineki; Kondo, Hiroyuki; Yasuhara, Akio; Hasebe, Kiyoshi

    2003-12-01

    A concentrated sulfuric acid dissolution technique and a GC method are described for the estimation of tributyl phosphate, tris(2-chloroethyl) phosphate, tris(chloropropyl) phosphate, tris(1,3-dichloro-2-propyl) phosphate, triphenyl phosphate and tris(butoxyethyl) phosphate in soft polyurethane foam. A soft polyurethane foam sample containing organophosphoric acid triesters was dissolved in concentrated sulfuric acid. The solution was added to water, where only the polyurethane was separated out. The pH of the solution was adjusted, and organophosphoric acid triesters were extracted with toluene. After purification, the compounds were determined by GC. The detection limits of the organophosphoric acid triesters were 0.3 - 0.9 microg g(-1). The recoveries of the organophosphoric acid triesters from a 0.05 g sample of soft polyurethane foam were 80.0 - 90.0%, when the spiked amounts were 0.25 - 1 microg. The compounds were detected from soft polyurethane foam at the level of 0.4 - 23.3 microg g(-1).

  10. Spectrophotometric method for fast quantification of ascorbic acid and dehydroascorbic acid in simple matrix for kinetics measurements.

    PubMed

    Gómez Ruiz, Braulio; Roux, Stéphanie; Courtois, Francis; Bonazzi, Catherine

    2016-11-15

    A simple, rapid and reliable method was developed for quantifying ascorbic (AA) and dehydroascorbic (DHAA) acids and validated in 20mM malate buffer (pH 3.8). It consists in a spectrophotometric measurement of AA, either directly on the solution added with metaphosphoric acid or after reduction of DHAA into AA by dithiothreitol. This method was developed with real time measurement of reactions kinetics in bulk reactors in mind, and was checked in terms of linearity, limits of detection and quantification, fidelity and accuracy. The linearity was found satisfactory on the range of 0-6.95mM with limits of detection and quantification of 0.236mM and 0.467mM, respectively. The method was found acceptable in terms of fidelity and accuracy with a coefficient of variation for repeatability and reproducibility below 6% for AA and below 15% for DHAA, and with a recovery range of 97-102% for AA and 88-112% for DHAA.

  11. Method comparison study for weak acid dissociation cyanide analysis.

    PubMed

    Evans, Joseph D; Thompson, Leslie; Clark, Patrick J; Beckman, Scott W

    2003-02-01

    Method comparison studies of two different methods for the analysis of weak acid dissociable (WAD) cyanide revealed analytical flaws and/or matrix interference problems with both procedures. EPA "draft" method 1677 using a Perstorp 3202 CN analyzer was compared to Standard Method 4500 CN I. It was discovered that the Perstorp analyzer produced more precise and more accurate results once appropriate and necessary procedural steps from the EPA draft method were modified. Comparison of these two methods, was based on "real world" samples collected from a mine-tailing solution. The mine-tailing solution contained high concentrations of cyanide and metals. Inconsistencies in method procedures were traced to sulfide interferences and high concentrations of WAD metals. Conclusions were based upon a large sample base collected from a mine site over a 90-day period.

  12. A Method for Detecting Positive Growth Autocorrelation without Marking Individuals

    PubMed Central

    Brooks, Mollie E.; McCoy, Michael W.; Bolker, Benjamin M.

    2013-01-01

    In most ecological studies, within-group variation is a nuisance that obscures patterns of interest and reduces statistical power. However, patterns of within-group variability often contain information about ecological processes. In particular, such patterns can be used to detect positive growth autocorrelation (consistent variation in growth rates among individuals in a cohort across time), even in samples of unmarked individuals. Previous methods for detecting autocorrelated growth required data from marked individuals. We propose a method that requires only estimates of within-cohort variance through time, using maximum likelihood methods to obtain point estimates and confidence intervals of the correlation parameter. We test our method on simulated data sets and determine the loss in statistical power due to the inability to identify individuals. We show how to accommodate nonlinear growth trajectories and test the effects of size-dependent mortality on our method's accuracy. The method can detect significant growth autocorrelation at moderate levels of autocorrelation with moderate-sized cohorts (for example, statistical power of 80% to detect growth autocorrelation ρ2 = 0.5 in a cohort of 100 individuals measured on 16 occasions). We present a case study of growth in the red-eyed tree frog. Better quantification of the processes driving size variation will help ecologists improve predictions of population dynamics. This work will help researchers to detect growth autocorrelation in cases where marking is logistically infeasible or causes unacceptable decreases in the fitness of marked individuals. PMID:24204620

  13. Vadose Zone Sampling Methods for Detection of Preferential Pesticides Transport

    NASA Astrophysics Data System (ADS)

    Peranginangin, N.; Richards, B. K.; Steenhuis, T. S.

    2003-12-01

    Leaching of agricultural applied chemicals through the vadose zone is a major cause for the occurrence of agrichemicals in groundwater. Accurate soil water sampling methods are needed to ensure meaningful monitoring results, especially for soils that have significant preferential flow paths. The purpose of this study was to assess the capability and the effectiveness of various soil water sampling methods in detecting preferential transport of pesticides in a strongly-structured silty clay loam (Hudson series) soil. Soil water sampling devices tested were wick pan and gravity pan lysimeters, tile lines, porous ceramic cups, and pipe lysimeters; all installed at 45 to105 cm depth below the ground surface. A reasonable worse-case scenario was tested by applying a simulated rain storm soon after pesticides were sprayed at agronomic rates. Herbicides atrazine (6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine) and 2,4-D (2,4-dichloro-phenoxyacetic acid) were chosen as model compounds. Chloride (KCl) tracer was used to determine spatial and temporal distribution of non-reactive solute and water as well as a basis for determining the retardation in pesticides movement. Results show that observed pesticide mobility was much greater than would be predicted by uniform flow. Under relatively high soil moisture conditions, gravity and wick pan lysimeters had comparably good collection efficiencies, whereas the wick samplers had an advantage over gravity driven sampler when the soil moisture content was below field capacity. Pipe lysimeters had breakthrough patterns that were similar to pan samplers. At small plot scale, tile line samplers tended to underestimate solute concentration because of water dilution around the samplers. The use of porous cup samplers performed poorly because of their sensitivity to local profile characteristics: only by chance can they intercept and sample the preferential flow paths that are critical to transport. Wick sampler had the least

  14. Acid pre-treatment method for in situ ore leaching

    DOEpatents

    Mallon, R.G.; Braun, R.L.

    1975-10-28

    An acid leaching method is described for the recovery of a desired element from a subterranean rubblized body of primary ore containing the element and also having associated therewith a carbonate mineral wherein the rubblized ore body is flooded with an aqueous acidic solution in order to release carbon dioxide from the associated carbonate mineral. After a substantial portion of the available carbon dioxide is released and removed from the ore body, as by venting to the atmosphere, an oxidizing gas is introduced into the flooded, rubblized ore to oxidize the ore and form an acid leach solution effective in the presence of the dissolved oxidizing gas to dissolve the ore and cause the desired element to go into solution. The leach solution is then circulated to the surface where the metal values are recovered therefrom.

  15. Fluorescent carbon nanodots for sensitive and selective detection of tannic acid in wines.

    PubMed

    Ahmed, Gaber Hashem Gaber; Laíño, Rosana Badía; Calzón, Josefa Angela García; García, Marta Elena Díaz

    2015-01-01

    Herein we describe an easy one step synthesis of carbon nanodots (C-dots) by thermal carbonization of 6-bromohexylboronic acid using two different amine compounds, polyethyleneglycol bis(3-aminopropyl (PEGA) and 1,2-aminopropane (DPA), at 180 °C in atmospheric oxygen. The as-synthesized C-dots were characterized by FTIR, HRTEM, NMR and fluorescence. The C-dots prepared using PEGA showed a strong emission at 440 nm with excitation at 362 nm. These C-dots exhibited analytical potential as sensing probes for tannic acid (TA) determination. pH effect, interferences, and analytical performance of the method were investigated. The method was found effective in the linear concentration range from 0.1 to 10 mg L(-1) TA achieving a limit of detection equal 0.018 mg L(-1) TA. The applicability of the method was demonstrated by direct measurements of TA in red and white wine samples. Validation of the method was achieved by spiking the wine samples with different standard TA concentrations obtaining recoveries in the range (90-112.5%). A probable mechanism by which TA quenched the C-dots fluorescence was proposed.

  16. Methods for detection of GMOs in food and feed.

    PubMed

    Marmiroli, Nelson; Maestri, Elena; Gullì, Mariolina; Malcevschi, Alessio; Peano, Clelia; Bordoni, Roberta; De Bellis, Gianluca

    2008-10-01

    This paper reviews aspects relevant to detection and quantification of genetically modified (GM) material within the feed/food chain. The GM crop regulatory framework at the international level is evaluated with reference to traceability and labelling. Current analytical methods for the detection, identification, and quantification of transgenic DNA in food and feed are reviewed. These methods include quantitative real-time PCR, multiplex PCR, and multiplex real-time PCR. Particular attention is paid to methods able to identify multiple GM events in a single reaction and to the development of microdevices and microsensors, though they have not been fully validated for application.

  17. Analysis of amino acid neurotransmitters from rat and mouse spinal cords by liquid chromatography with fluorescence detection.

    PubMed

    Şanlı, Nurullah; Tague, Sarah E; Lunte, Craig

    2015-03-25

    A RP-LC-FL detection method has been developed to identify and quantitate four amino acid neurotransmitters including glutamic acid, glycine, taurine and γ-aminobutyric acid in rat and mouse spinal cord tissue. 3-(4-carboxybenzoyl)-2-quinolinecarboxaldehyde (CBQCA) was employed for the derivatization of these neurotransmitters prior to RP-LC-FL analysis. Different parameters which influenced separation and derivatization were optimized. Under optimum conditions, linearity was achieved within the concentration ranges of 0.50-50.00 μM for all analytes with correlation coefficients from 0.9912 to 0.9997. The LODs ranged from 0.03 μM to 0.06 μM. The proposed method has been successfully applied to the determination of amino acid neurotransmitters in biological samples such as rat and mouse spinal cord with satisfactory recoveries.

  18. [Molecular Detection Methods for Vibrio parahaemolyticus in Seafood].

    PubMed

    Nishio, Tomohiro; Ohtsuka, Kayoko; Oda, Midori; Sugiyama, Kanji; Hara-Kudo, Yukiko

    2015-07-01

    To detect Vibrio parahaemolyticus in seafood, we evaluated efficient combinations of molecular methods with DNA extraction methods using heat extraction and alkaline heat extraction, and PCR, real-time PCR and loop-mediated isothermal amplification (LAMP) assays were performed targeting V parahaemolyticus species-specific genes (tlh and rpoD) and pathogenic factors genes (tdh and trh). The species-specific genes were detected in all combinations of two strains (a tdh * trh1-positive strain and a trh2-positive strain), two kinds of shellfish (oyster and bloody clams) and molecular methods with tlh-real time PCR or rpoD-LAMP assays with DNA of alkaline heat extraction at 85-145cfu/test level. tdh was detected in both seafoods with real time PCR assay with DNA of heat extraction at 85cfu/test level, and detected with the LAMP and real time PCR assays with DNA of alkaline heat extraction at 85cfu/test level. Detection of both trh1 and trh2 with the PCR assay with DNA of alkaline heat extraction was comparatively high though trh2 was detected with the LAMP assay with DNA of alkaline heat extraction at 145cfu/test level. It, however, is necessary to investigate more sensitive trh-detection methods. In this study, the results indicated that tlh-real time PCR or rpoD-LAMP, tdh-real time PCR and tdh-LAMP assays with DNA of alkaline heat extraction are relatively-sensitive methods to detect V. parahaemolyticus in seafood.

  19. Evaluation of Anomaly Detection Method Based on Pattern Recognition

    NASA Astrophysics Data System (ADS)

    Fontugne, Romain; Himura, Yosuke; Fukuda, Kensuke

    The number of threats on the Internet is rapidly increasing, and anomaly detection has become of increasing importance. High-speed backbone traffic is particularly degraded, but their analysis is a complicated task due to the amount of data, the lack of payload data, the asymmetric routing and the use of sampling techniques. Most anomaly detection schemes focus on the statistical properties of network traffic and highlight anomalous traffic through their singularities. In this paper, we concentrate on unusual traffic distributions, which are easily identifiable in temporal-spatial space (e.g., time/address or port). We present an anomaly detection method that uses a pattern recognition technique to identify anomalies in pictures representing traffic. The main advantage of this method is its ability to detect attacks involving mice flows. We evaluate the parameter set and the effectiveness of this approach by analyzing six years of Internet traffic collected from a trans-Pacific link. We show several examples of detected anomalies and compare our results with those of two other methods. The comparison indicates that the only anomalies detected by the pattern-recognition-based method are mainly malicious traffic with a few packets.

  20. A Bayesian Outbreak Detection Method for Influenza-Like Illness

    PubMed Central

    García, Yury E.; Christen, J. Andrés; Capistrán, Marcos A.

    2015-01-01

    Epidemic outbreak detection is an important problem in public health and the development of reliable methods for outbreak detection remains an active research area. In this paper we introduce a Bayesian method to detect outbreaks of influenza-like illness from surveillance data. The rationale is that, during the early phase of the outbreak, surveillance data changes from autoregressive dynamics to a regime of exponential growth. Our method uses Bayesian model selection and Bayesian regression to identify the breakpoint. No free parameters need to be tuned. However, historical information regarding influenza-like illnesses needs to be incorporated into the model. In order to show and discuss the performance of our method we analyze synthetic, seasonal, and pandemic outbreak data. PMID:26425552

  1. AquaLite, a bioluminescent label for immunoassay and nucleic acid detection: quantitative analyses at the attomol level

    NASA Astrophysics Data System (ADS)

    Smith, David F.; Stults, Nancy L.

    1996-04-01

    AquaLiteR is a direct, bioluminescent label capable of detecting attomol levels of analyte in clinical immunoassays and assays for the quantitative measurement of nucleic acids. Bioluminescent immunoassays (BIAs) require no radioisotopes and avoid complex fluorescent measurements and many of the variables of indirect enzyme immunoassays (EIAs). AquaLite, a recombinant form of the photoprotein aequorin from a bioluminescent jellyfish, is coupled directly to antibodies to prepare bioluminescent conjugates for assay development. When the AquaLite-antibody complex is exposed to a solution containing calcium ions, a flash of blue light ((lambda) max equals 469 nm) is generated. The light signal is measured in commercially available luminometers that simultaneously inject a calcium solution and detect subattomol photoprotein levies in either test tubes or microtiter plates. Immunometric or 'sandwich' type assays are available for the quantitative measurement of human endocrine hormones and nucleic acids. The AquaLite TSH assay can detect 1 attomol of thyroid stimulating hormone (TSH) in 0.2 mL of human serum and is a useful clinical tool for diagnosing hyperthyroid patients. AquaLite-based nucleic acid detection permits quantifying attomol levels of specific nucleic acid markers and represents possible solution to the difficult problem of quantifying the targets of nucleic acid amplification methods.

  2. Ultra-sensitive detection of zinc oxide nanowires using a quartz crystal microbalance and phosphoric acid DNA

    NASA Astrophysics Data System (ADS)

    Jang, Kuewhan; You, Juneseok; Park, Chanhoo; Park, Hyunjun; Choi, Jaeyeong; Choi, Chang-Hwan; Park, Jinsung; Lee, Howon; Na, Sungsoo

    2016-09-01

    Recent advancements of nanomaterials have inspired numerous scientific and industrial applications. Zinc oxide nanowires (ZnO NWs) is one of the most important nanomaterials due to their extraordinary properties. However, studies performed over the past decade have reported toxicity of ZnO NWs. Therefore, there has been increasing demand for effective detection of ZnO NWs. In this study, we propose a method for the detection of ZnO NW using a quartz crystal microbalance (QCM) and DNA probes. The detection method is based on the covalent interaction between ZnO NWs and the phosphoric acid group of single-stranded DNA (i.e., linker DNA), and DNA hybridization between the linker DNA and the probe DNA strand on the QCM electrode. Rapid, high sensitivity, in situ detection of ZnO NWs was demonstrated for the first time. The limit of detection was 10-4 μg ml-1 in deionized water, which represents a sensitivity that is 100000 times higher than the toxic ZnO NW concentration level. Moreover, the selectivity of the ZnO NW detection method was demonstrated by comparison with other types of nanowires and the method was able to detect ZnO NWs in tap water sensitively even after stored for 14 d in a refrigerator. The performance of our proposed method was sufficient to achieve detection of ZnO NW in the ‘real-world’ environment.

  3. Design, synthesis, and characterization of a protein sequencing reagent yielding amino acid derivatives with enhanced detectability by mass spectrometry.

    PubMed Central

    Aebersold, R.; Bures, E. J.; Namchuk, M.; Goghari, M. H.; Shushan, B.; Covey, T. C.

    1992-01-01

    We report the design, chemical synthesis, and structural and functional characterization of a novel reagent for protein sequence analysis by the Edman degradation, yielding amino acid derivatives rapidly detectable at high sensitivity by ion-evaporation mass spectrometry. We demonstrate that the reagent 3-[4'(ethylene-N,N,N-trimethylamino)phenyl]-2-isothiocyanate is chemically stable and shows coupling and cyclization/cleavage yields comparable to phenylisothiocyanate, the standard reagent in chemical sequence analysis, under conditions typically encountered in manual or automated sequence analysis. Amino acid derivatives generated with this reagent were detectable by ion-evaporation mass spectrometry at the subfemtomole sensitivity level at a pace of one sample per minute. Furthermore, derivatives were identified by their mass, thus permitting the rapid and highly sensitive determination of the molecular nature of modified amino acids. Derivatives of amino acids with acidic, basic, polar, or hydrophobic side chains were reproducibly detectable at comparable sensitivities. The polar nature of the reagent required covalent immobilization of polypeptides prior to automated sequence analysis. This reagent, used in automated sequence analysis, has the potential for overcoming the limitations in sensitivity, speed, and the ability to characterize modified amino acid residues inherent in the chemical sequencing methods that are currently used. PMID:1304351

  4. Biosensing platform for the detection of uric acid based on graphene quantum dots and G-quadruplex/hemin DNAzyme.

    PubMed

    Cai, Nan; Tan, Lu; Li, Yan; Xia, Tingting; Hu, Tianyu; Su, Xingguang

    2017-05-01

    In this paper, a label-free biosensing platform for fluorescence detection of uric acid was designed on the peroxidase-mimicking activities of G-quadruplex/hemin DNAzyme and the introduction of caffeic acid. Uric acid could be decomposed by uricase and then produced hydrogen peroxide and allantoin. We thus successfully achieved the indirect detection of uric acid by monitoring the concentration of hydrogen peroxide. The G-quadruplex/hemin DNAzyme could act as peroxidase and decompose the hydrogen peroxide into hydroxyl radicals at room temperature. Due to the strong oxidizing of hydroxyl radicals, caffeic acid was converted to corresponding quinone, thus leading to fluorescence quenching of GQDs. Under the optimized experimental conditions, the quenched fluorescence intensity was linearly relative to the concentration of uric acid, ranging from 2 μM to 300 μM with a detection limit of 500 nM. The applicability of proposed method was further proved with satisfactory results in human serum and urine samples.

  5. [Detection of mycobacteria tuberculosis in patients with urogenital tuberculosis by PCR method].

    PubMed

    Dochviri, T Z; Katsitadze, V A; Khosiashvili, G Z; Chigogidze, T G

    2005-02-01

    The study was carried out in hospital patients as well as in outpatients at the National Centre of Tuberculosis and Lung Diseases of Georgia (2002-2004). The group consisting of 32 patients with tuberculosis of urogenital system has been studied (newly detected forms). Except clinical laboratory, culture and X-ray contrast methods, two additional methods were used in testing of this group of patients. The examination of their urine, at the same time, was carried out by the Polymerase Chain Reaction method in order to detect Kochi bacillus and by three-time bacterioscopy of urine for acid resistant bacteria. Mycobacterium tuberculosis in urine has been detected in 26 (81,25%) patients by PCR method, and by urine bacterioscopy--acid fast bacilli (AFB+) in 18 (56,25%) patients. The histo-morphological investigation of specimens obtained by surgery confirmed the TB diagnosis in all patients. This study on patients suspected of Tuberculosis of genital-urinary system gives us an opportunity to update the diagnostic algorithm by including the modern molecular methods. This algorithm will help in timely detection of Tuberculosis, in selection of adequate therapy and in prevention of the further progression of the disease.

  6. Standardizing methylation method during phospholipid fatty acid analysis to profile soil microbial communities.

    PubMed

    Chowdhury, Taniya Roy; Dick, Richard P

    2012-02-01

    Phospholipid fatty acid (PLFA) as biomarkers, is widely used to profile microbial communities in environmental samples. However, PLFA extraction and derivatization protocols are not standardized and have widely varied among published studies. Specifically investigators have used either HCl/MeOH or KOH/MeOH or both for the methylation step of PLFA analysis, without justification or research to support either one. It seems likely that each method could have very different outcomes and conclusions for PLFA based studies. Therefore, the objective of this study was to determine the effect of catalyst type for methylation on detecting PLFAs and implications for interpreting microbial profiling in soil. Fatty acid samples extracted from soils obtained from a wetland, an intermittently flooded site, and an adjacent upland site were subjected to HCl/MeOH or KOH/MeOH catalyzed methylation procedures during PLFA analyses. The methylation method using HCl/MeOH resulted in significantly higher concentrations of most PLFAs than the KOH/MeOH method. Another important outcome was that fatty acids with a methyl group (18:1ω,7c 11Me, TBSA 10Me 18:0, 10Me 18:0, 17:0 10Me and 16:0 10Me being an actinomycetes biomarker) could not be detected by HCl/MeOH catalyzed methylation but were found in appreciable concentrations with KOH/MeOH method. From our results, because the HCl/MeOH method did not detect the fatty acids containing methyl groups that could strongly influence the microbial community profile, we recommend that the KOH/MeOH catalyzed transesterification method should become the standard procedure for PLFA profiling of soil microbial communities.

  7. Selective fluorescent detection of aspartic acid and glutamic acid employing dansyl hydrazine dextran conjugate.

    PubMed

    Nasomphan, Weerachai; Tangboriboonrat, Pramuan; Tanapongpipat, Sutipa; Smanmoo, Srung

    2014-01-01

    Highly water soluble polymer (DD) was prepared and evaluated for its fluorescence response towards various amino acids. The polymer consists of dansyl hydrazine unit conjugated into dextran template. The conjugation enhances higher water solubility of dansyl hydrazine moiety. Of screened amino acids, DD exhibited selective fluorescence quenching in the presence of aspartic acid (Asp) and glutamic acid (Glu). A plot of fluorescence intensity change of DD against the concentration of corresponding amino acids gave a good linear relationship in the range of 1 × 10(-4) M to 25 × 10(-3) M. This establishes DD as a potential polymeric sensor for selective sensing of Asp and Glu.

  8. Detection of non-protein amino acids in the presence of protein amino acids. II.

    NASA Technical Reports Server (NTRS)

    Shapshak, P.; Okaji, M.

    1972-01-01

    Studies conducted with the JEOL 5AH amino acid analyzer are described. This instrument makes possible the programming of the chromatographic process. Data are presented showing the separations of seventeen non-protein amino acids in the presence of eighteen protein amino acids. It is pointed out that distinct separations could be obtained in the case of a number of chemically similar compounds, such as ornithine and lysine, N-amidino alanine and arginine, and iminodiacetic acid and S-carboxymethyl cysteine and aspartic acid.

  9. Quantitative and discriminative analysis of nucleic acid samples using luminometric nonspecific nanoparticle methods

    NASA Astrophysics Data System (ADS)

    Pihlasalo, S.; Mariani, L.; Härmä, H.

    2016-03-01

    Homogeneous simple assays utilizing luminescence quenching and time-resolved luminescence resonance energy transfer (TR-LRET) were developed for the quantification of nucleic acids without sequence information. Nucleic acids prevent the adsorption of a protein to europium nanoparticles which is detected as a luminescence quenching of europium nanoparticles with a soluble quencher or as a decrease of TR-LRET from europium nanoparticles to the acceptor dye. Contrary to the existing methods based on fluorescent dye binding to nucleic acids, equal sensitivities for both single- (ssDNA) and double-stranded DNA (dsDNA) were measured and a detection limit of 60 pg was calculated for the quenching assay. The average coefficient of variation was 5% for the quenching assay and 8% for the TR-LRET assay. The TR-LRET assay was also combined with a nucleic acid dye selective to dsDNA in a single tube assay to measure the total concentration of DNA and the ratio of ssDNA and dsDNA in the mixture. To our knowledge, such a multiplexed assay is not accomplished with commercially available assays.Homogeneous simple assays utilizing luminescence quenching and time-resolved luminescence resonance energy transfer (TR-LRET) were developed for the quantification of nucleic acids without sequence information. Nucleic acids prevent the adsorption of a protein to europium nanoparticles which is detected as a luminescence quenching of europium nanoparticles with a soluble quencher or as a decrease of TR-LRET from europium nanoparticles to the acceptor dye. Contrary to the existing methods based on fluorescent dye binding to nucleic acids, equal sensitivities for both single- (ssDNA) and double-stranded DNA (dsDNA) were measured and a detection limit of 60 pg was calculated for the quenching assay. The average coefficient of variation was 5% for the quenching assay and 8% for the TR-LRET assay. The TR-LRET assay was also combined with a nucleic acid dye selective to dsDNA in a single tube

  10. Simultaneous determination of 4-hydroxyphenyl lactic acid, 4-hydroxyphenyl acetic acid, and 3-4-hydroxyphenyl propionic acid in human urine by Ultra high performance liquid chromatography with fluorescence detection.

    PubMed

    Yang, Yongli; Liu, Fan; Wan, Yiqun

    2017-03-27

    A simple and reliable method was established for simultaneous determination of 4-hydroxyphenyl acetic acid, 4-hydroxyphenyl lactic acid, and 3-4-hydroxyphenyl propionic acid in human urine by high-performance liquid chromatography with fluorescence detection. Solid-phase extraction was used to eliminate the interferences in urine. The separation of three analytes was achieved using a C18 column and a mobile phase formed by a 95:5 v/v mixture of 50 mmol/L ammonium acetate buffer at pH 6.8 that contained 5 mmol/L tetrabutyl ammonium bromide and acetonitrile. Under the optimized conditions, the detection limits of 4-hydroxyphenyl acetic acid, 4-hydroxyphenyl lactic acid, and 3-4-hydroxyphenyl propionic acid were 4.8 × 10(-3) , 8.80 × 10(-3) , and 9.00 × 10(-3) mg/L, respectively, and the recoveries were in the range of 85.0-120.0% with relative standard deviations of 1.5-3.1%. This method was used to analyze urine samples from breast cancer patients, healthy people and postsurgery breast cancer patients. Significant differences of urinary levels of 4-hydroxyphenyl acetic acid and 4-hydroxyphenyl lactic acid could be found between the breast cancer patients group and other two groups. No effect of age and sex was observed on the urinary levels of 4-hydroxyphenyl acetic acid and 4-hydroxyphenyl lactic acid. This method might be helpful for cancer biomarkers discovery in urine. This article is protected by copyright. All rights reserved.

  11. Methods and systems for remote detection of gases

    DOEpatents

    Johnson, Timothy J

    2012-09-18

    Novel systems and methods for remotely detecting at least one constituent of a gas via infrared detection are provided. A system includes at least one extended source of broadband infrared radiation and a spectrally sensitive receiver positioned remotely from the source. The source and the receiver are oriented such that a surface of the source is in the field of view of the receiver. The source includes a heating component thermally coupled to the surface, and the heating component is configured to heat the surface to a temperature above ambient temperature. The receiver is operable to collect spectral infrared absorption data representative of a gas present between the source and the receiver. The invention advantageously overcomes significant difficulties associated with active infrared detection techniques known in the art, and provides an infrared detection technique with a much greater sensitivity than passive infrared detection techniques known in the art.

  12. Methods and systems for remote detection of gases

    DOEpatents

    Johnson, Timothy J.

    2007-11-27

    Novel systems and methods for remotely detecting at least one constituent of a gas via infrared detection are provided. A system includes at least one extended source of broadband infrared radiation and a spectrally sensitive receiver positioned remotely from the source. The source and the receiver are oriented such that a surface of the source is in the field of view of the receiver. The source includes a heating component thermally coupled to the surface, and the heating component is configured to heat the surface to a temperature above ambient temperature. The receiver is operable to collect spectral infrared absorption data representative of a gas present between the source and the receiver. The invention advantageously overcomes significant difficulties associated with active infrared detection techniques known in the art, and provides an infrared detection technique with a much greater sensitivity than passive infrared detection techniques known in the art.

  13. Transistor-based particle detection systems and methods

    SciTech Connect

    Jain, Ankit; Nair, Pradeep R.; Alam, Muhammad Ashraful

    2015-06-09

    Transistor-based particle detection systems and methods may be configured to detect charged and non-charged particles. Such systems may include a supporting structure contacting a gate of a transistor and separating the gate from a dielectric of the transistor, and the transistor may have a near pull-in bias and a sub-threshold region bias to facilitate particle detection. The transistor may be configured to change current flow through the transistor in response to a change in stiffness of the gate caused by securing of a particle to the gate, and the transistor-based particle detection system may configured to detect the non-charged particle at least from the change in current flow.

  14. A novel homogenous detection method based on the self-assembled DNAzyme labeled DNA probes with SWNT conjugates and its application in detecting pathogen.

    PubMed

    Ding, Xinghua; Li, Hua; Deng, Le; Peng, Zhihui; Chen, Hui; Wang, Dan

    2011-07-15

    In this paper, a novel and cost-effective homogeneous detection method was constructed for the detection of genomic DNA and Staphylococcus aureus (S. aureus), based on the noncovalent assembly of DNAzyme-labeled detection probe and single-walled carbon nanotubes (SWNTs). When the target genomic DNA and hemin was existed in the detection solution, the detection probe wrapped on the SWNTs by π-stacking interactions would keep away from SWNTs and form a DNAzyme-self-assembly construction. This DNAzyme construction could catalyze 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS²⁻) and generate a colored product which could lead to the absorbance changes. Hence, according to its catalyzed capacity, the DNAzyme construction could amplify the detection signal. The concentration of target DNA could be quantified by exploiting their optical absorption changes at 414 nm and the concentration limit of detection of the method was 30 nM. And this detection method detected S. aureus quantitatively. In addition, this work proved that the method obtain higher detection sensitivity compared with the method without SWNTs because of the protection profile of SWNTs towards the detection probe.

  15. A high-throughput multiplex method adapted for GMO detection.

    PubMed

    Chaouachi, Maher; Chupeau, Gaëlle; Berard, Aurélie; McKhann, Heather; Romaniuk, Marcel; Giancola, Sandra; Laval, Valérie; Bertheau, Yves; Brunel, Dominique

    2008-12-24

    A high-throughput multiplex assay for the detection of genetically modified organisms (GMO) was developed on the basis of the existing SNPlex method designed for SNP genotyping. This SNPlex assay allows the simultaneous detection of up to 48 short DNA sequences (approximately 70 bp; "signature sequences") from taxa endogenous reference genes, from GMO constructions, screening targets, construct-specific, and event-specific targets, and finally from donor organisms. This assay avoids certain shortcomings of multiplex PCR-based methods already in widespread use for GMO detection. The assay demonstrated high specificity and sensitivity. The results suggest that this assay is reliable, flexible, and cost- and time-effective for high-throughput GMO detection.

  16. Detection of forced oscillations in power systems with multichannel methods

    SciTech Connect

    Follum, James D.

    2015-09-30

    The increasing availability of high fidelity, geographically dispersed measurements in power systems improves the ability of researchers and engineers to study dynamic behaviors in the grid. One such behavior that is garnering increased attention is the presence of forced oscillations. Power system engineers are interested in forced oscillations because they are often symptomatic of the malfunction or misoperation of equipment. Though the resulting oscillation is not always large in amplitude, the root cause may be serious. In this report, multi-channel forced oscillation detection methods are developed. These methods leverage previously developed detection approaches based on the periodogram and spectral-coherence. Making use of geographically distributed channels of data is shown to improved detection performance and shorten the delay before an oscillation can be detected in the online environment. Results from simulated and measured power system data are presented.

  17. New Optical Methods for Liveness Detection on Fingers

    PubMed Central

    Dolezel, Michal; Vana, Jan; Brezinova, Eva; Yim, Jaegeol; Shim, Kyubark

    2013-01-01

    This paper is devoted to new optical methods, which are supposed to be used for liveness detection on fingers. First we describe the basics about fake finger use in fingerprint recognition process and the possibilities of liveness detection. Then we continue with introducing three new liveness detection methods, which we developed and tested in the scope of our research activities—the first one is based on measurement of the pulse, the second one on variations of optical characteristics caused by pressure change, and the last one is based on reaction of skin to illumination with different wavelengths. The last part deals with the influence of skin diseases on fingerprint recognition, especially on liveness detection. PMID:24151584

  18. Development of a generic microfluidic device for simultaneous detection of antibodies and nucleic acids in oral fluids.

    PubMed

    Chen, Zongyuan; Abrams, William R; Geva, Eran; de Dood, Claudia J; González, Jesús M; Tanke, Hans J; Niedbala, R Sam; Zhou, Peng; Malamud, Daniel; Corstjens, Paul L A M

    2013-01-01

    A prototype dual-path microfluidic device (Rheonix CARD) capable of performing simultaneously screening (antigen or antibody) and confirmatory (nucleic acid) detection of pathogens is described. The device fully integrates sample processing, antigen or antibody detection, and nucleic acid amplification and detection, demonstrating rapid and inexpensive "sample-to-result" diagnosis with performance comparable to benchtop analysis. For the chip design, a modular approach was followed allowing the optimization of individual steps in the sample processing process. This modular design provides great versatility accommodating different disease targets independently of the production method. In the detection module, a lateral flow (LF) protocol utilizing upconverting phosphor (UCP) reporters was employed. The nucleic acid (NA) module incorporates a generic microtube containing dry reagents. Lateral flow strips and PCR primers determine the target or disease that is diagnosed. Diagnosis of HIV infection was used as a model to investigate the simultaneous detection of both human antibodies against the virus and viral RNA. The serological result is available in less than 30 min, and the confirmation by RNA amplification takes another 60 min. This approach combines a core serological portable diagnostic with a nucleic acid-based confirmatory test.

  19. Indirect UV detection-ion-exclusion/cation-exchange chromatography of common inorganic ions with sulfosalicylic acid eluent.

    PubMed

    Kozaki, Daisuke; Mori, Masanobu; Nakatani, Nobutake; Arai, Kaori; Masuno, Tomoe; Koseki, Masakazu; Itabashi, Hideyuki; Tanaka, Kazuhiko

    2013-01-01

    Herein, we describe indirect UV detection-ion-exclusion/cation-exchange chromatography (IEC/CEC) on a weakly acidic cation-exchange resin in the H(+)-form (TSKgel Super IC-A/C) using sulfosalicylic acid as the eluent. The goal of the study was to characterize the peaks detected by UV detector. The peak directions of analyte ions in UV at 315 nm were negative because the molar absorbance coefficients of analyte anions and cations were lower than that of the sulfosalicylic acid eluent. Good chromatographic resolution and high signal-to-noise ratios of analyte ions were obtained for the separations performed using 1.1 mM sulfosalicylic acid and 1.5 mM 18-crown-6 as the eluent. The relative standard deviations (RSDs) of the peak areas ranged from 0.6 to 4.9%. Lower detection limits of the analytes were achieved using indirect UV detection at 315 nm (0.23 - 0.98 μM) than those obtained with conductometric detection (CD) (0.61 - 2.1 μM) under the optimized elution conditions. The calibration curves were linear in the range from 0.01 to 1.0 mM except for Cl(-), which was from 0.02 to 2.0 mM. The present method was successfully applied to determine common inorganic ions in a pond water sample.

  20. A review on detection methods used for foodborne pathogens

    PubMed Central

    Priyanka, B.; Patil, Rajashekhar K.; Dwarakanath, Sulatha

    2016-01-01

    Foodborne pathogens have been a cause of a large number of diseases worldwide and more so in developing countries. This has a major economic impact. It is important to contain them, and to do so, early detection is very crucial. Detection and diagnostics relied on culture-based methods to begin with and have developed in the recent past parallel to the developments towards immunological methods such as enzyme-linked immunosorbent assays (ELISA) and molecular biology-based methods such as polymerase chain reaction (PCR). The aim has always been to find a rapid, sensitive, specific and cost-effective method. Ranging from culturing of microbes to the futuristic biosensor technology, the methods have had this common goal. This review summarizes the recent trends and brings together methods that have been developed over the years. PMID:28139531