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Sample records for acid detection methods

  1. 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.

  2. 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.

  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. Rapid detection method for fusaric acid-producing species of Fusarium by PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fusaric acid is a mycotoxin produced by species of the fungus Fusarium and can act synergistically with other Fusarium toxins. In order to develop a specific detection method for fusaric acid-producing fungus, PCR prim¬ers were designed to amplify FUB10, a transcription factor gene in fusaric acid ...

  6. 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.

  7. New method and detection of high concentrations of monomethylarsonous acid detected in contaminated groundwater.

    PubMed

    McKnight-Whitford, Anthony; Chen, Baowei; Naranmandura, Hua; Zhu, Chen; Le, X Chris

    2010-08-01

    Monomethylarsonous acid (MMAIII) was detected in groundwater from a former herbicide production plant in the USA. The site has total arsenic concentrations up to thousands of mg/L, representing one of the most severe cases of arsenic contamination ever reported. Structure-specific detection of MMAIII, along with arsenite (AsIII), arsenate (AsV), monomethylarsonic acid (MMAV), and dimethylarsinic acid (DMAV), was achieved using liquid chromatography separation with electrospray ionization tandem mass spectrometry detection (HPLC-ESI-MS/MS). To enable the electrospray of MMAIII and AsIII, dimercaptosuccinic acid (DMSA) was used to derivatize these trivalent arsenicals online, so that their complexes with DMSA could be detected using negative ionization ESI-MS/MS. The presence of MMAIII was verified using high resolution mass spectrometry to measure accurate mass, tandem mass spectrometry to monitor fragmentation, and three different separation techniques to resolve arsenic species. The measured accurate mass of the suspected MMAIII compound in a groundwater sample was 122.9607+/-0.0003 amu, which was in good agreement with the theoretical value and that of the MMAIII standard. Simultaneous monitoring of AsO+ at m/z 91 and SO+ at m/z 48 using HPLC-ICPMS operating in dynamic reaction cell mode ruled out possible confounding from any sulfur-containing arsenic compound. The concentrations of MMAIII found in the groundwater samples from a contaminated site were as high as 3.9-274 mg/L, the highest ever observed in the environment.

  8. 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.

  9. 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.

  10. 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. PMID:27507479

  11. 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.

  12. 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.

  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. PMID:19619859

  14. A modified method for the detection of microbial proteases on agar plates using tannic acid.

    PubMed

    Saran, Saurabh; Isar, Jasmine; Saxena, Rajendra Kumar

    2007-06-10

    In routine assay for the screening of microbes producing proteases, 10% trichloroaceticacid (TCA) is flooded on the milk agar plates after inoculation and required incubation to precipitate the protein. However, the clarity of the hydrolyzed zone is not very sharp and distinct. We herein present an improved assay for detecting the presence of extracellular protease from microorganisms on agar plates. In this method 10% tannic acid is flooded on the milk agar plate (in place of, TCA) to observe the zone of hydrolysis. Tannic acid sharply increases the colour intensity of the plate, as it favours the precipitation of the unhydrolyzed protein in the plate, thereby improving the contrast between the intact zones and the enzymatic lyses zones of the substrate. Our results indicate that this method is useful to detect extracellular proteases produced by both fungi as well as bacteria. The method used in the present study is sensitive, and can be easily performed for screening of large number of microbial cultures. This is the first report on the use of tannic acid for the detection of microbial proteases.

  15. 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...

  16. 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...

  17. 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.

  18. 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. PMID:25575759

  19. 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.

  20. A reliable and inexpensive method of nucleic acid extraction for the PCR-based detection of diverse plant pathogens.

    PubMed

    Li, R; Mock, R; Huang, Q; Abad, J; Hartung, J; Kinard, G

    2008-12-01

    A reliable extraction method is described for the preparation of total nucleic acids from at least ten plant genera for subsequent detection of plant pathogens by PCR-based techniques. The method combined a modified CTAB (cetyltrimethylammonium bromide) extraction protocol with a semi-automatic homogenizer (FastPrep) instrument) for rapid sample processing and low potential of cross contamination. The method was applied to sample preparation for PCR-based detection of 28 different RNA and DNA viruses, six viroids, two phytoplasmas and two bacterial pathogens from a range of infected host plants including sweet potato, small fruits and fruit trees. The procedure is cost-effective and the qualities of the nucleic acid preparations are comparable to those prepared by commonly used commercial kits. The efficiency of the procedure permits processing of numerous samples and the use of a single nucleic acid preparation for testing both RNA and DNA genomes by PCR, making this an appealing method for testing multiple pathogens in certification and quarantine programs.

  1. 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). PMID:27393827

  2. 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.

  3. Evaluation of a facile method of template DNA preparation for PCR-based detection and typing of lactic acid bacteria.

    PubMed

    Singh, Atul Kumar; Ramesh, Aiyagari

    2009-08-01

    The objective of our investigation was to develop a convenient and reliable method of generating template DNA for routine PCR-based detection and typing of lactic acid bacteria (LAB). Template DNA extracted from Lactobacillus, Lactococcus, Pediococcus and Leuconostoc using a combination of urea, SDS and NaOH yielded amplicons of expected size in PCR with genus-specific primers. Apart from LAB, the proposed method could also be adopted to generate PCR-compatible template DNA from a number of Gram-positive and Gram-negative bacterial strains. DNA template prepared by the proposed method from various standard strains of Lactobacillus sp. also generated discriminating fingerprints with BOXA1R primer in rep-PCR. A significant finding of the investigation was that a comparable banding profile of LAB strains was obtained in rep-PCR using template DNA prepared by urea-SDS-NaOH method and a commercially available DNA isolation kit. This was further evidenced by high dice coefficient values obtained in the range of 81.8-96.7 when cluster analysis was performed by UPGAMA method. The application potential of this DNA extraction method for PCR-based direct detection of LAB in fermented food samples such as dahi, idli batter and salt-fermented cucumber was validated by detecting specific amplicons of LAB genera in the fermented samples. The applicability of the proposed template DNA extraction method was further substantiated when 29 bacteriocinogenic LAB strains (Bac+) previously detected in salt-fermented cucumber by PCR [Singh, A.K., Ramesh, A., 2008. Succession of dominant and antagonistic lactic acid bacteria in fermented cucumber: Insights from a PCR-based approach. Food. Microbiol. 25, 278-287] generated differentiating fingerprints in BOX element based rep-PCR and formed clusters with reference LAB strains. PMID:19465247

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

    PubMed

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

    2011-01-01

    According to the new KDIGO (Kidney Disease Improving Global Outcomes) 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 (37º, 45º, 60º, 70º and 80ºC) 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 60ºC and 70ºC. For technical reasons and according to the results of the present study, we recommended that, for an incubation time of 30 minutes, the reaction should be carried out at 60

  5. 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

  6. Evaluation of non-radioactive labelling and detection of deoxyribonucleic acids. Part Two: Colorigenic methods and comparison with chemiluminescent methods.

    PubMed

    Rihn, B; Bottin, M C; Coulais, C; Martinet, N

    1995-06-01

    The diagnosis of genetic infections and cancerous diseases is carried out more and more often at a molecular level using Southern's technique which is based on the use of 32P-labelled DNA. In order to circumvent the risks and rapid decrease in radioactivity associated with these latter techniques, new colorigenic methods have been developed. In this work, we describe the use of dTTP analogues (digoxigenin-dUTP and biotin-dUTP) for the labelling of probes and detection of target DNA. Using digoxigenin-11-dUTP, 0.1 aM of a 561 bp target DNA was detected by using a modified Southern procedure. The reliability and the high sensitivity of such methods make them a good tool for DNA investigation in research as well as in testing laboratories.

  7. 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. ...

  8. 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. PMID:24036075

  9. An optimized analytical method for the simultaneous detection of iodoform, iodoacetic acid, and other trihalomethanes and haloacetic acids in drinking water.

    PubMed

    Liu, Xiaolin; Wei, Xiao; Zheng, Weiwei; 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 (R(2) 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.

  10. 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.

  11. Development of HPLC method by UV-VIS detection for the quantification of phenolic acids in different Ocimum sanctum Linn. extracts.

    PubMed

    Shafqatullah; Khan, Rasool; Hassan, Waseem; Hussain, Arshad; Asadullah; Rehman, Khaliqur; Ali, Javid

    2014-09-01

    A simple and rapid chromatographic method has been developed for the simultaneous determination of five phenolic acids including Gallic acid, Chloroganic acid, Syringic acid, Benzoic acid and Vanillic acid by HPLC with UV-VIS detector. These Phenolic acids were separated by analytical column Intersil ODS-3 C18, a gradient elution system of ACN and acidified water solution with 1ml/min flow rate and quantified in a total run of 30 minutes at 210nm wavelength. In the quantitative analysis of these compounds showed good regression (0.995-0.999). The limit of detection [LOD] and limit of quantification [LOQ] of these compounds were in the range of 0.15-0.46 and 0.42-2.47 βg/mL. The average recoveries were between 95.8-103.1% and their RSD values were less than 3.34%. By the proposed method Gallic acid, Chloroganic acid and Syringic acid were found and quantified in Methanolic, Ethanolic and Acetonic extract of Ocimum sanctum Linn. leaves. While the two other phenolic acids benzoic acid and vanillic acid was not found in the extracts of Ocimum sanctum Linn. leaves. PMID:25176382

  12. 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. PMID:16176651

  13. 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.

  14. Fluorescence in situ Hybridization method using Peptide Nucleic Acid probes for rapid detection of Lactobacillus and Gardnerella spp.

    PubMed Central

    2013-01-01

    Background Bacterial vaginosis (BV) is a common vaginal infection occurring in women of reproductive age. It is widely accepted that the microbial switch from normal microflora to BV is characterized by a decrease in vaginal colonization by Lactobacillus species together with an increase of Gardnerella vaginalis and other anaerobes. Our goal was to develop and optimize a novel Peptide Nucleic Acid (PNA) Fluorescence in situ Hybridization assay (PNA FISH) for the detection of Lactobacillus spp. and G. vaginalis in mixed samples. Results Therefore, we evaluated and validated two specific PNA probes by using 36 representative Lactobacillus strains, 22 representative G. vaginalis strains and 27 other taxonomically related or pathogenic bacterial strains commonly found in vaginal samples. The probes were also tested at different concentrations of G. vaginalis and Lactobacillus species in vitro, in the presence of a HeLa cell line. Specificity and sensitivity of the PNA probes were found to be 98.0% (95% confidence interval (CI), from 87.8 to 99.9%) and 100% (95% CI, from 88.0 to 100.0%), for Lactobacillus spp.; and 100% (95% CI, from 92.8 to 100%) and 100% (95% CI, from 81.5 to 100.0%) for G. vaginalis. Moreover, the probes were evaluated in mixed samples mimicking women with BV or normal vaginal microflora, demonstrating efficiency and applicability of our PNA FISH. Conclusions This quick method accurately detects Lactobacillus spp. and G. vaginalis species in mixed samples, thus enabling efficient evaluation of the two bacterial groups, most frequently encountered in the vagina. PMID:23586331

  15. 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.

  16. 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.

  17. High-performance liquid chromatography-ultraviolet detection method for the simultaneous determination of typical biogenic amines and precursor amino acids. Applications in food chemistry.

    PubMed

    Mazzucco, Eleonora; Gosetti, Fabio; Bobba, Marco; Marengo, Emilio; Robotti, Elisa; Gennaro, Maria Carla

    2010-01-13

    A reversed-phase high-performance liquid chromatography (HPLC) method was developed for the simultaneous determination in food of biogenic amines and their precursor amino acids after a precolumn derivatization with dansyl chloride. The chromatographic conditions, selected to be suitable for mass spectrometry detection, were optimized through experimental design and artificial neural networks. The HPLC-UV method was validated by comparing the separation results with those obtained through a HPLC method, working under the same chromatographic conditions but employing mass spectrometry detection. The HPLC-UV method was then applied to the analysis of different food samples, namely, cheese, clams, salami, and beer. For all of the matrices, recoveries (relative standard deviation always <5%) always >92% were obtained. The results are discussed as a function of the total biogenic amine content and of the concentration ratio between amines and precursor amino acids.

  18. 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

  19. 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

  20. Multigrid contact detection method

    NASA Astrophysics Data System (ADS)

    He, Kejing; Dong, Shoubin; Zhou, Zhaoyao

    2007-03-01

    Contact detection is a general problem of many physical simulations. This work presents a O(N) multigrid method for general contact detection problems (MGCD). The multigrid idea is integrated with contact detection problems. Both the time complexity and memory consumption of the MGCD are O(N) . Unlike other methods, whose efficiencies are influenced strongly by the object size distribution, the performance of MGCD is insensitive to the object size distribution. We compare the MGCD with the no binary search (NBS) method and the multilevel boxing method in three dimensions for both time complexity and memory consumption. For objects with similar size, the MGCD is as good as the NBS method, both of which outperform the multilevel boxing method regarding memory consumption. For objects with diverse size, the MGCD outperform both the NBS method and the multilevel boxing method. We use the MGCD to solve the contact detection problem for a granular simulation system based on the discrete element method. From this granular simulation, we get the density property of monosize packing and binary packing with size ratio equal to 10. The packing density for monosize particles is 0.636. For binary packing with size ratio equal to 10, when the number of small particles is 300 times as the number of big particles, the maximal packing density 0.824 is achieved.

  1. 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. PMID:18023403

  2. 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.

  3. 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.

  4. Use of Dimedone-Based Chemical Probes for Sulfenic Acid Detection: Methods to Visualize and Identify Labeled Proteins

    PubMed Central

    Nelson, Kimberly J.; Klomsiri, Chananat; Codreanu, Simona G.; Soito, Laura; Liebler, Daniel C.; Rogers, LeAnn C.; Daniel, Larry W.; Poole, Leslie B.

    2013-01-01

    Reversible thiol modification is a major component of the modulation of cell-signaling pathways by reactive oxygen species. Hydrogen peroxide, peroxynitrite, or lipid hydroperoxides are all able to oxidize cysteines to form cysteine sulfenic acids; this reactive intermediate can be directly reduced to thiol by cellular reductants such as thioredoxin or further participate in disulfide bond formation with glutathione or cysteine residues in the same or another protein. To identify the direct protein targets of cysteine modification and the conditions under which they are oxidized, a series of dimedone-based reagents linked to affinity or fluorescent tags have been developed that specifically alkylate and trap cysteine sulfenic acids. In this chapter, we provide detailed methods using one of our biotin-tagged reagents, DCP-Bio1, to identify and monitor proteins that are oxidized in vitro and in vivo. Using streptavidin-linked agarose beads, this biotin-linked reagent can be used to affinity capture labeled proteins. Stringent washing of the beads prior to elution minimizes the contamination of the enriched material with unlabeled proteins through coimmunoprecipitation or nonspecific binding. In particular, we suggest including DTT in one of the washes to remove proteins covalently linked to biotinylated proteins through a disulfide bond, except in cases where these linked proteins are of interest. We also provide methods for targeted approaches monitoring cysteine oxidation in individual proteins, global approaches to follow total cysteine oxidation in the cell, and guidelines for proteomic analyses to identify novel proteins with redox sensitive cysteines. PMID:20513473

  5. [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.

  6. 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. PMID:26601859

  7. 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

  8. 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. PMID:25720597

  9. 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.

  10. Methods for Doping Detection.

    PubMed

    Ponzetto, Federico; Giraud, Sylvain; Leuenberger, Nicolas; Boccard, Julien; Nicoli, Raul; Baume, Norbert; Rudaz, Serge; Saugy, Martial

    2016-01-01

    Over the past few years, the World Anti-Doping Agency (WADA) has focused its efforts on detecting not only small prohibited molecules, but also larger endogenous molecules such as hormones, in the view of implementing an endocrinological module in the Athlete Biological Passport (ABP). In this chapter, the detection of two major types of hormones used for doping, growth hormone (GH) and endogenous anabolic androgenic steroids (EAASs), will be discussed: a brief historical background followed by a description of state-of-the-art methods applied by accredited anti-doping laboratories will be provided and then current research trends outlined. In addition, microRNAs (miRNAs) will also be presented as a new class of biomarkers for doping detection. PMID:27348309

  11. 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.).

  12. [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.

  13. Disposable terbium (III) salicylate complex imprinted membrane using solid phase surface fluorescence method for fast separation and detection of salicylic acid in pharmaceuticals and human urine.

    PubMed

    Huang, Jianxiang; Hu, Yufei; Hu, Yuling; Li, Gongke

    2013-03-30

    In this work, a simple, low cost, selective and sensitive complex imprinted membrane (CIM) for solid-phase fluorescent detection was developed with terbium (III) salicylate as complex template. Terbium-sensitized luminescence was employed for monitoring salicylic acid (SA) based on the fluorescence enhancement effect of benzoic acid derivatives on lanthanide ion Tb (III). The resulting CIM showed good fluorescent response and high selectivity towards SA with Tb as pivot in protic solvents, while demonstrating better analytical performance than the controlled membranes. The optimized adsorption time was 10 min, indicating rapid kinetics of the imprinted membrane. The linear response of CIM to SA was from 0.20 to 10mg/L with limit of detection (LOD) of 0.040 mg/L. The prepared CIM was successfully applied to the analysis of salicylic acid in pharmaceuticals and spiked human urine with recoveries of 80.6%-88.1%. The analytical results of the proposed method were in good agreement with those obtained by high performance liquid chromatography (HPLC) method, indicating that the developed membrane has acceptable practicability for fast determination of SA in real samples.

  14. 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

  15. 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

  16. Detection of Escherichia coli O157 by Peptide Nucleic Acid Fluorescence In Situ Hybridization (PNA-FISH) and Comparison to a Standard Culture Method

    PubMed Central

    Almeida, C.; Sousa, J. M.; Rocha, R.; Cerqueira, L.; Fanning, S.; Azevedo, N. F.

    2013-01-01

    Despite the emergence of non-O157 Shiga toxin-producing Escherichia coli (STEC) infections, E. coli serotype O157 is still the most commonly identified STEC in the world. It causes high morbidity and mortality and has been responsible for a number of outbreaks in many parts of the world. Various methods have been developed to detect this particular serotype, but standard bacteriological methods remain the gold standard. Here, we propose a new peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) method for the rapid detection of E. coli O157. Testing on 54 representative strains showed that the PNA probe is highly sensitive and specific to E. coli O157. The method then was optimized for detection in food samples. Ground beef and unpasteurized milk samples were artificially contaminated with E. coli O157 concentrations ranging from 1 × 10−2 to 1 × 102 CFU per 25 g or ml of food. Samples were then preenriched and analyzed by both the traditional bacteriological method (ISO 16654:2001) and PNA-FISH. The PNA-FISH method performed well in both types of food matrices with a detection limit of 1 CFU/25 g or ml of food samples. Tests on 60 food samples have shown a specificity value of 100% (95% confidence interval [CI], 82.83 to 100), a sensitivity of 97.22% (95% CI, 83.79 to 99.85%), and an accuracy of 98.33% (CI 95%, 83.41 to 99.91%). Results indicate that PNA-FISH performed as well as the traditional culture methods and can reduce the diagnosis time to 1 day. PMID:23934486

  17. Optimized culturing and nucleic acid-based methods for the detection of Salmonella enterica in poultry environments.

    PubMed

    Schultz, J; Jarquin, R; Ricke, S C; Hanning, I

    2012-11-01

    The main objective of this trial was to set up a series of assays following quantified inoculation of Salmonella samples in 2 initial enrichment methods to ultimately determine the most effective and fastest detection method for recovery of Salmonella in a poultry environment matrix. Samples were randomly split into 2 different containers containing either buffered peptone water (BPW) + yeast extract, or tetrathionate broth (TT) with added iodine and Brilliant Green solution 0.1%. A frozen stock Salmonella culture was thawed and serially diluted 10-fold to inoculate 100 µL of the dilution into the enriched samples. The samples were incubated at 42 and 37°C, respectively, for 24 h and secondarily enriched in modified semi-solid Rappaport Vassiliadis (MSRV) incubated at 42°C. All samples then were reincubated under the same conditions. After secondary enrichment, the samples were streaked onto Chromogenic agar/ XLT4 bi-plates and incubated under the same conditions. After initial inoculation and each 24-h incubation, a portion of the enriched samples was analyzed using a real-time PCR assay. The results of this trial indicate that recovery of Salmonella in a culture-based assay may be enhanced by up to 3 logs by using the TT as the initial enrichment media compared with BPW. The incorporation of MSRV as a secondary cultural selective media after the TT gave the best recovery of Salmonella. These data indicate that considerable time can be saved by using TT as an initial media for Salmonella recovery. PMID:23091129

  18. 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. PMID:23993512

  19. 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.

  20. 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...

  1. A sensitive GC-EIMS method for simultaneous detection and quantification of JWH-018 and JWH-073 carboxylic acid and hydroxy metabolites in urine.

    PubMed

    Paul, Buddha D; Bosy, Thomas

    2015-04-01

    Synthetic cannabinoids, including JWH-018 and JWH-073, belong to a class of aminoalkylindoles (AAIs) that are smoked to produce an effect similar to tetrahydrocannabinol. Compounds in this class are often collectively known as 'Spice'. After ingestion, these compounds are extensively metabolized to their hydroxy and carboxylic acid metabolites. During forensic analysis, detection of these metabolites in urine is an indication of past exposure to the parent compounds. The analytical process involved hydrolysis of conjugated metabolites by glucuronidase, solvent extraction, derivatization by trifluoroacetic anhydride and hexafluoroisopropanol and GC-EIMS detection. Identification of the unknown was based on the criteria of GC retention time within ±2% and mass spectral ion ratio within ±20% of that of a standard. Deuterated internal standards of the carboxylic acid metabolites were used for quantification. The acid (JWH-018-COOH, JWH-073-COOH) and hydroxy (JWH-018-OH, JWH-073-OH) metabolites were linear over the concentration range of 0.1-10 and 0.2-10 ng/mL, respectively, with a correlation coefficient-square, R(2) > 0.999 (N = 5). Extraction recoveries of the metabolites were within 79 and 87%. The method was applied to 17 urine specimens collected as part of a military law enforcement investigation. Nine of the specimens tested positive for one or more of the metabolites. When the procedure was extended to screen other AAI compounds, two of the specimens were found to contain JWH-210, JWH-250 (JWH-302 or JWH-201) and JWH-250 (C4 isomers). The GC-EIMS method presented here was found to be suitable for detecting JWH-018 and JWH-073 metabolites and other AAI compounds in urine.

  2. A convenient HPLC method for detection of okadaic acid analogs as 9-anthrylmethyl esters with automated sample cleanup by column switching.

    PubMed

    Uchida, Hajime; Watanabe, Ryuichi; Matsushima, Ryoji; Uchida, Naoyuki; Nagai, Hiroshi; Kamio, Michiya; Murata, Masakazu; Yasumoto, Takeshi; Suzuki, Toshiyuki

    2014-01-01

    A convenient HPLC-fluorometric detection (FLD) method for okadaic acid (OA) analogs as 9-anthrylmethyl esters was developed with the addition of column switching to simplify and automate cleanup. Methanol extracts of shellfish were first treated to hydrolyze OA esters and then reacted with 9-anthryldiazomethane (ADAM). ADAM derivatives of OA and dinophysistoxin-1 (DTX1) were subsequently determined by HPLC-FLD following automated column-switching cleanup. The LOD (S/N = 3) and LOQ (S/N = 10) of OA and DTX1 obtained from bivalves fortified with toxin in our method were approximately 2.6 and 8.6 ng/g whole meat, respectively. The recoveries of OA and DTX1 at all fortification levels of bivalve extracts ranged from 90 to 113%, with RSD values of 0.9-9.9%. The new method is applicable to the routine monitoring of OA analogs as an inexpensive and convenient alternative to HPLC/MS.

  3. Detection of nucleic acids by multiple sequential invasive cleavages

    SciTech Connect

    Hall, J.G.; Lyamichev, V.I.; Mast, A.L.; Brow, M.A.D.

    1999-11-30

    The present invention relates to methods 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. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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

  9. A rapid and simple method to determine the specific activities of serotonin, 5-hydroxyindoleacetic acid, and 5-hydroxytryptophan in brain by HPLC with electrochemical detection

    SciTech Connect

    Bernstein, M.J.; Shea, P.A.

    1982-01-01

    The specific activities of 5-hydroxytryptophan (5-HTP), serotonin (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) have been determined in the brain of rats by HPLC using electrochemical detection. The method allows, from a single sample, the simultaneous measurement of all three compounds and collection of each peak for radioactivity determinations. Five male Wistar rats were injected i.v. with 2.0 mCi/kg of DL-5-hydroxy-(G-/sup 3/H)tryptophan (2.6 Ci/mmol) and 30 min later the animals were killed by near freezing. Whole brains were removed and homogenized in an acid medium. The content of 5-HTP, 5-HT, and 5-HIAA were determined by HPLC. Each peak of interest was immediately collected after detection in scintillation vials by use of a small dead space detector (TL-9A, Bioanalytical Systems, Inc.). The amounts of radioactivity were determined and specific activities calculated from the results. A second chromatography system (TLC) was used to check the authenticity and purity of compounds separated by the HPLC.

  10. Human nails metabolite analysis: A rapid and simple method for quantification of uric acid in human fingernail by high-performance liquid chromatography with UV-detection.

    PubMed

    Li, Xi-Ling; Li, Gao; Jiang, Ying-Zi; Kang, Dongzhou; Jin, Cheng Hua; Shi, Qing; Jin, Toufeng; Inoue, Koichi; Todoroki, Kenichiro; Toyo'oka, Toshimasa; Min, Jun Zhe

    2015-10-01

    A rapid and simple analytical method for the quantification of uric acid (UA) in human fingernails by high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection is described. UA was extracted from human fingernail samples at 90°C for 20min, then separated on an Inertsil ODS-2 column (250×4.6mm I.D., 5.0μm, GL Sciences) by isocratic elution using methanol: 74mM phosphate buffer (pH 2.2) 2:98 (v/v). An UV detector was used to monitor at 284nm. The results indicated that under optimized measurement conditions results were achieved within 8.0min, and a good linearity was achieved from the calibration curves (r(2)>0.9999) in the range of 1.0-10000ng; the limit of detection (S/N=3) was 2.0pg, the inter-day and intra-day assay precisions were all less than 0.46% and the mean recoveries (%) of the uric acid spiked in the human fingernail were 101.95%. The amounts of UA in the fingernails of healthy volunteers were determined.

  11. Broth Microdilution Method To Detect Extended-Spectrum β-Lactamases and AmpC β-Lactamases in Enterobacteriaceae Isolates by Use of Clavulanic Acid and Boronic Acid as Inhibitors ▿

    PubMed Central

    Jeong, Seok Hoon; Song, Wonkeun; Kim, Jae-Seok; Kim, Han-Sung; Lee, Kyu Man

    2009-01-01

    This study was designed to evaluate the performance of the broth microdilution (BMD) method to detect production of extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamases in Enterobacteriaceae by using clavulanic acid (CA) and boronic acid (BA) as ESBL and AmpC β-lactamase inhibitors, respectively. A total of 100 clinical isolates of Enterobacteriaceae were analyzed. Mueller-Hinton broth containing serial twofold dilutions of cefotaxime (CTX), ceftazidime (CAZ), aztreonam (ATM), or cefepime (FEP) with or without either or both CA and BA was prepared. An eightfold or greater decrease in the MIC of CTX, CAZ, ATM, or FEP in the presence of CA and BA was considered a positive result for ESBL and plasmid-mediated AmpC β-lactamase (PABL), respectively. In tests with CA, expanded-spectrum β-lactams containing BA (CTX-BA, CAZ-BA, ATM-BA, and FEP-BA) showed higher positive rates in detecting ESBL producers than those without BA. The combination of CTX- and CAZ-based BMD tests with CA and BA showed sensitivity and specificity of 100% for the detection of ESBLs and PABLs. The BMD testing could be applicable for routine use in commercially available semiautomated systems for the detection of ESBLs and PABLs in Enterobacteriaceae. PMID:19710269

  12. 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. PMID:19733195

  13. Validation of a Fluorescence In Situ Hybridization Method Using Peptide Nucleic Acid Probes for Detection of Helicobacter pylori Clarithromycin Resistance in Gastric Biopsy Specimens

    PubMed Central

    Cerqueira, Laura; Fernandes, Ricardo M.; Ferreira, Rui M.; Oleastro, Mónica; Carneiro, Fátima; Brandão, Catarina; Pimentel-Nunes, Pedro; Dinis-Ribeiro, Mário; Figueiredo, Céu; Keevil, Charles W.; Vieira, Maria J.

    2013-01-01

    Here, we evaluated a previously established peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) method as a new diagnostic test for Helicobacter pylori clarithromycin resistance detection in paraffin-embedded gastric biopsy specimens. Both a retrospective study and a prospective cohort study were conducted to evaluate the specificity and sensitivity of a PNA-FISH method to determine H. pylori clarithromycin resistance. In the retrospective study (n = 30 patients), full agreement between PNA-FISH and PCR-sequencing was observed. Compared to the reference method (culture followed by Etest), the specificity and sensitivity of PNA-FISH were 90.9% (95% confidence interval [CI], 57.1% to 99.5%) and 84.2% (95% CI, 59.5% to 95.8%), respectively. In the prospective cohort (n = 93 patients), 21 cases were positive by culture. For the patients harboring clarithromycin-resistant H. pylori, the method showed sensitivity of 80.0% (95% CI, 29.9% to 98.9%) and specificity of 93.8% (95% CI, 67.7% to 99.7%). These values likely represent underestimations, as some of the discrepant results corresponded to patients infected by more than one strain. PNA-FISH appears to be a simple, quick, and accurate method for detecting H. pylori clarithromycin resistance in paraffin-embedded biopsy specimens. It is also the only one of the methods assessed here that allows direct and specific visualization of this microorganism within the biopsy specimens, a characteristic that allowed the observation that cells of different H. pylori strains can subsist in very close proximity in the stomach. PMID:23596234

  14. 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.

  15. 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.

  16. 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.

  17. Method for nucleic acid isolation using supercritical fluids

    DOEpatents

    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.

  18. 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.

  19. 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.

  20. Method for detecting leaks

    SciTech Connect

    Baillie, L.A.; Senese, F.J.

    1988-09-27

    This patent describes a method for determining leaks in a storage tank containing a liquid. The method consists of: locating a float means initially at a depth in the liquid approximately equal to V/sub L//S/sub L/ where V/sub L/ is the volume of the liquid in the tank and S/sub L/ is free surface area of the liquid in the tank; saturating vapor above the liquid in the tank with vapor of the liquid by atomizing so as to fill the vapor above the liquid with small drops of liquid or coating surfaces of the tank above the liquid with a thin layer of the liquid so as to hasten saturation of the vapor above the liquid, so that temperature variation of the liquid in the tank does not change the depth at which the float means floats in the liquid; and determining changes of location of the float means in the tank.

  1. 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.

  2. 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.

  3. Local method for detecting communities

    NASA Astrophysics Data System (ADS)

    Bagrow, James P.; Bollt, Erik M.

    2005-10-01

    We propose a method of community detection that is computationally inexpensive and possesses physical significance to a member of a social network. This method is unlike many divisive and agglomerative techniques and is local in the sense that a community can be detected within a network without requiring knowledge of the entire network. A global application of this method is also introduced. Several artificial and real-world networks, including the famous Zachary karate club, are analyzed.

  4. 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

  5. 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.

  6. 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

  7. Well acidizing compositions and methods

    SciTech Connect

    Swanson, B. L.

    1980-12-23

    Gelled acidic compositions suitable for matrix acidizing or fracture acidizing of subterranean formations are provided comprising water, a water-dispersible polymeric viscosifier such as a polymer of acrylamide, an acid, and a polyphenolic material such as lignite.

  8. 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.

  9. A Rapid Method for Determination of the Main Conjugated Linoleic Acid Precursors (C18:2 n-6 and C18:3 n-3) in Forage by Capillary Zone Electrophoresis with Ultraviolet Detection Using Gas Chromatography with Flame Ionization Detection as a Comparative Method.

    PubMed

    de Jesus Coelho Castro, Renata; Sobrinho, Fausto Souza; Sundfeld da Gama, Marco Antônio; Takabayashi Sato, Renata; Lavorato Lima, Larissa; Leal de Oliveira, Marcone Augusto

    2015-01-01

    A rapid method has been proposed for determination of the main conjugated linoleic acid precursors such as linoleic (C18:2 n-6) and linolenic (C18:3 n-3) acids in forages by capillary zone electrophoresis (CZE) with direct UV detection at 200 nm. Among the fatty acids found in forages, C18:2 n-6 and C18:3 n-3 have received particular attention due to their roles as precursors for the synthesis of conjugated linoleic acid, a class of health-enhancing compounds that is predominantly found in dairy products. The electrolyte background consisted of 12.0 mmol/L tetraborate buffer (pH 9.2) added to 12.0 mmol/L Brij 35®, 17% acetonitrile, and 33% methanol. Under the optimized conditions, the baseline separation of C18:2 n-6 and C18:3 n-3 was achieved within 4 min. The CZE-UV method was compared to GC with a flame ionization detector, which is the American Oil Chemists' Society (AOCS 996.06) official method for fatty acid analysis. The methods did not show any evidence of significant differences within 95% confidence interval (P>0.05). The CZE-UV method was successfully applied to the analysis of 80 genotypes of Brachiaria ruzizienses clones submitted to a genetic improvement program in agricultural research. PMID:26651572

  10. 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.

  11. Nucleic acid detection using G-quadruplex amplification methodologies.

    PubMed

    Roembke, Benjamin T; Nakayama, Shizuka; Sintim, Herman O

    2013-12-15

    In the last decade, there has been an explosion in the use of G-quadruplex labels to detect various analytes, including DNA/RNA, proteins, metals and other metabolites. In this review, we focus on strategies for the detection of nucleic acids, using G-quadruplexes as detection labels or as enzyme labels that amplify detection signals. Methods to detect other analytes are briefly mentioned. We highlight various strategies, including split G-quadruplex, hemin-G-quadruplex conjugates, molecular beacon G-quadruplex or inhibited G-quadruplex probes. The tandem use of G-quadruplex labels with various DNA-modifying enzymes, such as polymerases (used for rolling circle amplification), exonucleases and endonucleases, is also discussed. Some of the detection modalities that are discussed in this review include fluorescence, colorimetric, chemiluminescence, and electrochemical methods.

  12. 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.

  13. 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.

  14. 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.

  15. Detection of Respiratory Viruses by Molecular Methods

    PubMed Central

    Mahony, James B.

    2008-01-01

    Summary: Clinical laboratories historically diagnose seven or eight respiratory virus infections using a combination of techniques including enzyme immunoassay, direct fluorescent antibody staining, cell culture, and nucleic acid amplification tests. With the discovery of six new respiratory viruses since 2000, laboratories are faced with the challenge of detecting up to 19 different viruses that cause acute respiratory disease of both the upper and lower respiratory tracts. The application of nucleic acid amplification technology, particularly multiplex PCR coupled with fluidic or fixed microarrays, provides an important new approach for the detection of multiple respiratory viruses in a single test. These multiplex amplification tests provide a sensitive and comprehensive approach for the diagnosis of respiratory tract infections in individual hospitalized patients and the identification of the etiological agent in outbreaks of respiratory tract infection in the community. This review describes the molecular methods used to detect respiratory viruses and discusses the contribution that molecular testing, especially multiplex PCR, has made to our ability to detect respiratory viruses and to increase our understanding of the roles of various viral agents in acute respiratory disease. PMID:18854489

  16. 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.

  17. 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.

  18. 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.

  19. Detecting acid precipitation impacts on lake water quality

    NASA Astrophysics Data System (ADS)

    Loftis, Jim C.; Taylor, Charles H.

    1989-09-01

    The United States Environmental Protection Agency is planning to expand its long-term monitoring of lakes that are sensitive to acid deposition effects. Effective use of resources will require a careful definition of the statistical objectives of monitoring, a network design which balances spatial and temporal coverage, and a sound approach to data analysis. This study examines the monitoring objective of detecting trends in water quality for individual lakes and small groups of lakes. Appropriate methods of trend analysis are suggested, and the power of trend detection under seasonal (quarterly) sampling is compared to that of annual sampling. The effects of both temporal and spatial correlation on trend detection ability are described.

  20. 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.

  1. Nucleic acid detection systems for enteroviruses.

    PubMed Central

    Rotbart, H A

    1991-01-01

    The enteroviruses comprise nearly 70 human pathogens responsible for a wide array of diseases including poliomyelitis, meningitis, myocarditis, and neonatal sepsis. Current diagnostic tests for the enteroviruses are limited in their use by the slow growth, or failure to grow, of certain serotypes in culture, the antigenic diversity among the serotypes, and the low titer of virus in certain clinical specimens. Within the past 6 years, applications of molecular cloning techniques, in vitro transcription vectors, automated nucleic acid synthesis, and the polymerase chain reaction have resulted in significant progress toward nucleic acid-based detection systems for the enteroviruses that take advantage of conserved genomic sequences across many, if not all, serotypes. Similar approaches to the study of enteroviral pathogenesis have already produced dramatic advances in our understanding of how these important viruses cause their diverse clinical spectra. PMID:1649002

  2. 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...

  3. Immunofluorescence detection methods using microspheres

    NASA Astrophysics Data System (ADS)

    Szurdoki, Ferenc; Michael, Karri L.; Agrawal, Divya; Taylor, Laura C.; Schultz, Sandra L.; Walt, David R.

    1999-01-01

    Microsphere-based immunoassays were devised for compounds of agricultural and biomedical interest (e.g., digoxin, theophylline, and zearalenone). Commercially available microspheres with surface functional groups for chemical derivatization were used as solid carriers. After immobilizing the target substances, the surface of the haptenized microspheres was blocked by a protein to reduce aspecific binding. Competitive immunoassays were performed using the functionalized microspheres and antibodies labeled with horseradish peroxidase. Immunofluorescence signal amplification was achieved by enzyme-catalyzed reporter deposition (CARD). An epifluorescence microscope, a CCD camera interfaced with a computer, and microscopy image analysis software were employed for quantitative detection of fluorescent light emitted from individual microspheres. Integration of several such immunoassays and application of an optical encoding method enabled multianalyte determination. These immunoassays can also be utilized in an immunosensor array format. This immunoarray format could facilitate miniaturization and automation of multianalyte immunoassays.

  4. LABORATORY DETECTION OF THIOCYANIC ACID HSCN

    SciTech Connect

    Bruenken, S.; Yu, Z.; Gottlieb, C. A.; McCarthy, M. C.; Thaddeus, P. E-mail: cgottlieb@cfa.harvard.ed E-mail: pthaddeus@cfa.harvard.ed

    2009-12-01

    The rotational spectrum of thiocyanic acid HSCN, a highly polar isomer of the well-known astronomical molecule isothiocyanic acid HNCS, has been measured in two radio bands: in the centimeter-wave band by Fourier transform microwave spectroscopy in a molecular beam, and in the millimeter-wave band by long-path absorption spectroscopy in a low-pressure glow discharge. Twelve spectroscopic constants were derived from more than 60 a-type rotational transitions between 11 and 346 GHz with J up to 30 and K{sub a} <= 6, including seven centimeter-wave transitions with resolved hyperfine structure. With these constants the rotational spectrum in the K{sub a} = 0 and K{sub a} = 1 ladders-those most likely to be observed in space-can now be calculated up to 400 GHz with formal uncertainties of less than 0.2 km s{sup -1} in equivalent radial velocity. Thiocyanic acid was recently identified in Sgr B2 by Halfen et al. following the laboratory measurements, and there is possible evidence for it in cold dark clouds, with the implication that HSCN may be detectable in many galactic sources.

  5. Methods for preparation of deuterated amino acids

    SciTech Connect

    Pshenichnikova, A.B.; Karnaukhova, E.N.; Zvonkova, E.N.

    1995-03-01

    The current state and prospects for the use of amino acids labeled with stable isotopes are considered. Methods for the preparation of deuterated amino acids, including synthetic, chemicoenzymatic, and biosynthetic ones, and deuterium exchange reactions are summarized. Problems in the preparation of optically pure amino acids are discussed. 120 refs., 15 figs.

  6. Safer staining method for acid fast bacilli.

    PubMed Central

    Ellis, R C; Zabrowarny, L A

    1993-01-01

    To develop a method for staining acid fast bacilli which excluded highly toxic phenol from the staining solution. A lipophilic agent, a liquid organic detergent, LOC High Studs, distributed by Amway, was substituted. The acid fast bacilli stained red; nuclei, cytoplasm, and cytoplasmic elements stained blue on a clear background. These results compare very favourably with acid fast bacilli stained by the traditional method. Detergents are efficient lipophilic agents and safer to handle than phenol. The method described here stains acid fast bacilli as efficiently as traditional carbol fuchsin methods. LOC High Suds is considerably cheaper than phenol. Images PMID:7687254

  7. Safer staining method for acid fast bacilli.

    PubMed

    Ellis, R C; Zabrowarny, L A

    1993-06-01

    To develop a method for staining acid fast bacilli which excluded highly toxic phenol from the staining solution. A lipophilic agent, a liquid organic detergent, LOC High Studs, distributed by Amway, was substituted. The acid fast bacilli stained red; nuclei, cytoplasm, and cytoplasmic elements stained blue on a clear background. These results compare very favourably with acid fast bacilli stained by the traditional method. Detergents are efficient lipophilic agents and safer to handle than phenol. The method described here stains acid fast bacilli as efficiently as traditional carbol fuchsin methods. LOC High Suds is considerably cheaper than phenol.

  8. Optimized Dispersive Liquid-Liquid Microextraction Method and High Performance Liquid Chromatography with Ultraviolet Detection for Simultaneous Determination of Sorbic and Benzoic Acids and Evaluation of Contamination of These Preservatives in Iranian Foods.

    PubMed

    Javanmardi, Fardin; Arefhosseini, Seyyed Rafie; Ansarin, Masood; Nemati, Mahboob

    2015-01-01

    A rapid, simple, and sensitive dispersive liquid-liquid microextraction procedure followed by HPLC-UV was applied to determine the benzoate and sorbate in foods. The method was optimized for some variables including extraction solvent type and volume, dispersing solvent type and volume, and the effects of salt and pH. Optimum conditions were determined as follows: sample volume, 5 mL; extraction solvent (chloroform) volume, 250 μL; disperser solvent (acetone) volume, 1.2 mL; NaCl amount, 0.75 g/5 mL at pH 4. Sixty samples were analyzed, including 15 doogh, 15 fruit juice, 15 cookie, and 15 tomato paste; benzoic acid was detected in 57 samples (95%) at levels up to 448.1 μg/mL and sorbic acid in 31 samples (51.6%) at levels up to 1369 μg/mL. Under the optimum experimental conditions, the LOD and LOQ were determined as 0.1 and 0.5 μg/mL for benzoate and 0.08 and 0.3 μg/mL for sorbate, respectively. The results showed that these preservatives are commonly used at high levels in yogurt drinks (dooghs) and cookies. Also, the concentration of benzoic acid that was detected in the tomato paste and fruit juice samples was low but may affect children and sensitive persons.

  9. Affinity sensor using 3-aminophenylboronic acid for bacteria detection.

    PubMed

    Wannapob, Rodtichoti; Kanatharana, Proespichaya; Limbut, Warakorn; Numnuam, Apon; Asawatreratanakul, Punnee; Thammakhet, Chongdee; Thavarungkul, Panote

    2010-10-15

    Boronic acid that can reversibly bind to diols was used to detect bacteria through its affinity binding reaction with diol-groups on bacterial cell walls. 3-aminophenylboronic acid (3-APBA) was immobilized on a gold electrode via a self-assembled monolayer. The change in capacitance of the sensing surface caused by the binding between 3-APBA and bacteria in a flow system was detected by a potentiostatic step method. Under optimal conditions the linear range of 1.5×10(2)-1.5×10(6) CFU ml(-1) and the detection limit of 1.0×10(2) CFU ml(-1) was obtained. The sensing surface can be regenerated and reused up to 58 times. The method was used for the analysis of bacteria in several types of water, i.e., bottled, well, tap, reservoir and wastewater. Compared with the standard plate count method, the results were within one standard deviation of each other. The proposed method can save both time and cost of analysis. The electrode modified with 3-APBA would also be applicable to the detection of other cis-diol-containing analytes. The concept could be extended to other chemoselective ligands, offering less expensive and more robust affinity sensors for a wide range of compounds.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. A sensitive and efficient method for determination of N-acetylhexosamines and N-acetylneuraminic acid in breast milk and milk-based products by high-performance liquid chromatography via UV detection and mass spectrometry identification.

    PubMed

    Chuanxiang, Wu; Lian, Xia; Lijie, Liu; Fengli, Qu; Zhiwei, Sun; Xianen, Zhao; Jinmao, You

    2016-02-01

    A sensitive and efficient method of high performance liquid chromatography using 1-(2-naphthyl)-3-methyl-5-pyrazolone (NMP) as pre-column derivatization reagent coupled with UV detection (HPLC-UV) and online mass spectrometry identification was established for determination of the most common N-Acetylhexosamines (N-acetyl-d-glucosamine (GlcNAc) and N-acetyl-d-galactosamine (GalNAc)) and N-acetylneuraminic acid (Neu5Ac). In order to obtain the highest liberation level of the three monosaccharides without destruction of Neu5Ac or conversion of GlcNAc/GalNAc to GlcN/GalN in the hydrolysis procedure, the pivotal parameters affecting the liberation of N-acetylhexosamines/Neu5Ac from sample were investigated with response surface methodology (RSM). Under the optimized condition, maximum yield was obtained. The effects of key parameters on derivatization, separation and detection were also investigated. At optimized conditions, three monosaccharides were labeled fast and entirely, and all derivatives exhibited a good baseline resolution and high detection sensitivity. The developed method was linear over the calibration range 0.25-12μM, with R(2)>0.9991. The detection limits of the method were between 0.48 and 2.01pmol. Intra- and inter-day precisions for the three monosaccharides (GlcNAc, GalNAc and Neu5Ac) were found to be in the range of 3.07-4.02% and 3.69-4.67%, respectively. Individual monosaccharide recovery from spiked milk was in the range of 81%-97%. The sensitivity of the method, the facility of the derivatization procedure and the reliability of the hydrolysis conditions suggest the proposed method has a high potential for utilization in routine trace N-acetylhexosamines and Neu5Ac analysis in biological samples.

  15. 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.

  16. Nucleic acid detection technologies and marker molecules in bacterial diagnostics.

    PubMed

    Scheler, Ott; Glynn, Barry; Kurg, Ants

    2014-05-01

    There is a growing need for quick and reliable methods for microorganism detection and identification worldwide. Although traditional culture-based technologies are trustworthy and accurate at a relatively low cost, they are also time- and labor-consuming and are limited to culturable bacteria. Those weaknesses have created a necessity for alternative technologies that are capable for faster and more precise bacterial identification from medical, food or environmental samples. The most common current approach is to analyze the nucleic acid component of analyte solution and determine the bacterial composition according to the specific nucleic acid profiles that are present. This review aims to give an up-to-date overview of different nucleic acid target sequences and respective analytical technologies.

  17. 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.

  18. 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.

  19. 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.

  20. Nucleic acid sequence detection using multiplexed oligonucleotide PCR

    SciTech Connect

    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.

  1. 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

  2. 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.

  3. 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.

  4. Challenges with fats and fatty acid methods.

    PubMed

    Palmquist, D L; Jenkins, T C

    2003-12-01

    The content and chemical nature of lipids in feedstuffs is heterogeneous. It has long been known that ether extraction by the Weende procedure inadequately characterizes the fat content of feedstuffs, yet it remains the official method. Diethyl ether (or hexanes that are often used) extracts significant amounts of nonnutritive, nonsaponifiable lipids from forages, and often incompletely extracts lipids of nutritional value, especially fatty acids present as salts of divalent cations. Preextraction hydrolysis of insoluble fatty acid salts with acid releases these fatty acids, and this step is included in the official procedure for certain feedstuffs in the United Kingdom; however, acid hydrolysis increases analysis time and decreases precision. Acid hydrolysis also causes confusion as to the proper definition of the fat content of feedstuffs. A preferred method of fat analysis determines the total fatty acid concentration in feed samples by converting fatty acid salts, as well as the acyl components in all lipid classes, such as triacylglycerols, phospholipids, and sphingolipids, to methyl esters using a simple, direct one-step esterification procedure. Fatty acid methyl esters are then quantified by GLC, which provides information on both fatty acid quantity and profile in a single analysis. Adjustments in conditions and reagents may be necessary to overcome difficulty in quantitatively preparing esters from certain types of fatty acids and their derivatives in commercial fat supplements. After correction for glycerol content, analysis of oils by this procedure provides information on the content of nonsaponifiable material, such as chlorophyll, waxes, and indigestible polymers formed from heat- or oxidatively damaged fats. The correct description of feedstuffs for energy value of fats is the content of total fatty acids. PMID:14677882

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

    PubMed

    Gerasimova, Yulia V; Kolpashchikov, Dmitry M

    2014-09-01

    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.

  6. 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.

  7. Origin of D-amino acids detected in the acid hydrolysates of purified Escherichia coli β-galactosidase.

    PubMed

    Miyamoto, Tetsuya; Sekine, Masae; Ogawa, Tetsuhiro; Hidaka, Makoto; Homma, Hiroshi; Masaki, Haruhiko

    2015-12-10

    In previous report, we detected D-amino acids in the acid hydrolysates of purified recombinant β-galactosidase. Here, we employed a deuterium-hydrogen exchange method to discriminate innate D-amino acids from those generated during hydrolytic incubation. After hydrolysis of β-galactosidase in DCl/D2O, amino acids were derivatized with NBD-F and separated on a reverse-phase column, followed by liquid chromatography-tandem mass spectrometry equipped with a chiral column. Our results show an absence of innate D-amino acid residues in the protein and suggest that the protein undergoes isomerization during a very early stage of hydrolytic incubation.

  8. Nucleic Acids for Ultra-Sensitive Protein Detection

    PubMed Central

    Janssen, Kris P. F.; Knez, Karel; Spasic, Dragana; Lammertyn, Jeroen

    2013-01-01

    Major advancements in molecular biology and clinical diagnostics cannot be brought about strictly through the use of genomics based methods. Improved methods for protein detection and proteomic screening are an absolute necessity to complement to wealth of information offered by novel, high-throughput sequencing technologies. Only then will it be possible to advance insights into clinical processes and to characterize the importance of specific protein biomarkers for disease detection or the realization of “personalized medicine”. Currently however, large-scale proteomic information is still not as easily obtained as its genomic counterpart, mainly because traditional antibody-based technologies struggle to meet the stringent sensitivity and throughput requirements that are required whereas mass-spectrometry based methods might be burdened by significant costs involved. However, recent years have seen the development of new biodetection strategies linking nucleic acids with existing antibody technology or replacing antibodies with oligonucleotide recognition elements altogether. These advancements have unlocked many new strategies to lower detection limits and dramatically increase throughput of protein detection assays. In this review, an overview of these new strategies will be given. PMID:23337338

  9. 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.

  10. Variation-Tolerant Capture and Multiplex Detection of Nucleic Acids: Application to Detection of Microbes

    PubMed Central

    Öhrmalm, Christina; Eriksson, Ronnie; Jobs, Magnus; Simonson, Magnus; Strømme, Maria; Bondeson, Kåre; Herrmann, Björn; Melhus, Åsa

    2012-01-01

    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. PMID:22814465

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. Method for Trace Oxygen Detection

    NASA Technical Reports Server (NTRS)

    Man, Kim Fung (Inventor); Boumsellek, Said (Inventor); Chutjian, Ara (Inventor)

    1997-01-01

    Trace levels of molecular oxygen are measured by introducing a gas containing the molecular oxygen into a target zone, and impacting the molecular oxygen in the target zone with electrons at the O(-) resonant energy level for dissociative electron attachment to produce O(-) ions. Preferably, the electrons have an energy of about 4 to about 10 eV. The amount of O(-) ions produced is measured, and is correlated with the molecular oxygen content in the target zone. The technique is effective for measuring levels of oxygen below 50 ppb. and even less than 1 ppb. The amount of O(-) can be measured in a quadrupole mass analyzer. Best results are obtained when the electrons have an energy of about 6 to about 8 eV. and preferably about 6.8 eV. The method can be used for other species by selecting the appropriate electron energy level.

  17. 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.

  18. 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.).

  19. 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.). PMID:21391500

  20. 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. PMID:19899783

  1. 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.

  2. Bioluminescent bioreporter integrated circuit detection methods

    DOEpatents

    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.

  3. 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...

  4. 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.

  5. Methods for the detection of marine toxins

    SciTech Connect

    Wekell, M.M.; Manger, R.M.; Hadley, S.W.; Hungerford, J.M.

    1995-12-01

    Toxic materials have been dumped into the seas from waste streams and other pollutant sources such as runoff, etc. For protection of public health, it is essential that rapid, reliable and simple methods exist to detect marine toxins in seafoods. In addition, it is necessary to develop methods requiring a minimum of test material. Pure standards for many of the marine toxins are scarce. Reduced sample requirements extend the utility of detection methods in research and forensic applications as well. In the past, there was much reliance on the animal bioassay; however, this dependence hopefully will be reduced as newer instrumental techniques (chromatographic, mass spectrometric, electrophoretic), biochemical (immunochemical, receptor site assay), and cell bioassay methods are developed with a higher degree of precision and specificity. It is beneficial that a multiplicity of methods be available to detect marine toxins in seafoods. Each method has unique advantages and disadvantages.

  6. Analytic sequential methods for detecting network intrusions

    NASA Astrophysics Data System (ADS)

    Chen, Xinjia; Walker, Ernest

    2014-05-01

    In this paper, we propose an analytic sequential methods for detecting port-scan attackers which routinely perform random "portscans" of IP addresses to find vulnerable servers to compromise. In addition to rigorously control the probability of falsely implicating benign remote hosts as malicious, our method performs significantly faster than other current solutions. We have developed explicit formulae for quick determination of the parameters of the new detection algorithm.

  7. 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.

  8. 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. PMID:27664615

  9. 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.

  10. 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...

  11. Coverage methods for early groundwater contamination detection.

    PubMed

    Nunes, Luís Miguel; da Conceição Cunha, Maria; Ribeiro, Luís

    2013-05-01

    A method based on space-filling coverage designs to optimize groundwater monitoring networks for plume detection and quantification is proposed. Space-filling objective functions are then compared with more classical functions. The method was applied to a hypothetical case-study with 160 candidate locations, resulting in final optimal design monitoring networks with 40 locations. Results show that the method is superior to those based strictly on the probability of contamination detection for quantifying maximum and mean values. In the light of these results fractal properties of space-filling coverage methods and of simulated annealing are also discussed.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. Adaptive sequential methods for detecting network intrusions

    NASA Astrophysics Data System (ADS)

    Chen, Xinjia; Walker, Ernest

    2013-06-01

    In this paper, we propose new sequential methods for detecting port-scan attackers which routinely perform random "portscans" of IP addresses to find vulnerable servers to compromise. In addition to rigorously control the probability of falsely implicating benign remote hosts as malicious, our method performs significantly faster than other current solutions. Moreover, our method guarantees that the maximum amount of observational time is bounded. In contrast to the previous most effective method, Threshold Random Walk Algorithm, which is explicit and analytical in nature, our proposed algorithm involve parameters to be determined by numerical methods. We have introduced computational techniques such as iterative minimax optimization for quick determination of the parameters of the new detection algorithm. A framework of multi-valued decision for detecting portscanners and DoS attacks is also proposed.

  17. 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.

  18. 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.

  19. 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

  20. 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

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

    PubMed

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

    2016-07-01

    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

  2. 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.

  3. Inner Crack Detection Method for Cantilever Beams

    NASA Astrophysics Data System (ADS)

    Li, Zheng; Zhang, Wei; Li, Yixuan; Su, Xianyue

    2008-02-01

    In this paper, continuous wavelet transform has been performed to extract the inner crack information from the guided waves in cantilever beams, and the location and size of crack can be detected exactly. Considering its best time-frequency property, Gabor continuous wavelet transform is employed to analyze the complicated flexible wave signals in cantilever beam, which is inspirited by an impact on the free end. Otherwise, in order to enhance the sensitivity of detection for some small cracks, an improved method is discussed. Here, both computational and experimental methods are carried out for comparing the influence of different crack location in beam. Therefore, the method proposed can be expected to expand to a powerful damage detection method in a broad engineering application.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. 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.

  13. 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.

  14. Detection of exogenous citric acid in fruit juices by stable isotope ratio analysis.

    PubMed

    Jamin, Eric; Martin, Frédérique; Santamaria-Fernandez, Rebeca; Lees, Michèle

    2005-06-29

    A new method has been developed for measuring the D/H ratio of the nonexchangeable sites of citric acid by isotope ratio mass spectrometry (IRMS). Pure citric acid is transformed into its calcium salt and subsequently analyzed by pyrolysis-IRMS. The citric acid isolated from authentic fruit juices (citrus, pineapple, and red fruits) systematically shows higher D/H values than its nonfruit counterpart produced by fermentation of various sugar sources. The discrimination obtained with this simplified method is similar to that obtained previously by applying site specific isotopic fractionation-nuclear magnetic resonance (SNIF-NMR) to an ester derivative of citric acid. The combination of carbon 13 and deuterium measurements of extracted citric acid is proposed as a routine method for an optimum detection of exogenous citric acid in all kinds of fruit juices.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. Novel methods for detecting buried explosive devices

    NASA Astrophysics Data System (ADS)

    Kercel, Stephen W.; Burlage, Robert S.; Patek, David R.; Smith, Cyrus M.; Hibbs, Andrew D.; Rayner, Timothy J.

    1997-07-01

    Oak Ridge National Laboratory and Quantum Magnetics, Inc. 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, one 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.

  20. A Locked Nucleic Acid Probe Based on Selective Salt-Induced Effect Detects Single Nucleotide Polymorphisms

    PubMed Central

    Zhang, Jing; Wu, Huizhe; Chen, Qiuchen; Zhao, Pengfei; Zhao, Haishan; Yao, Weifan; Wei, Minjie

    2015-01-01

    Detection of single based genetic mutation by using oligonucleotide probes is one of the common methods of detecting single nucleotide polymorphisms at known loci. In this paper, we demonstrated a hybridization system which included a buffer solution that produced selective salt-induced effect and a locked nucleic acid modified 12 nt oligonucleotide probe. The hybridization system is suitable for hybridization under room temperature. By using magnetic nanoparticles as carriers for PCR products, the SNPs (MDR1 C3435T/A) from 45 volunteers were analyzed, and the results were consistent with the results from pyrophosphoric acid sequencing. The method presented in this paper differs from the traditional method of using molecular beacons to detect SNPs in that it is suitable for research institutions lacking real-time quantitative PCR detecting systems, to detect PCR products at room temperature. PMID:26347880

  1. 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.

  2. 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.

  3. 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.

  4. Method and apparatus for detecting an analyte

    DOEpatents

    Allendorf, Mark D.; Hesketh, Peter J.

    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.

  5. Improved astigmatic focus error detection method

    NASA Technical Reports Server (NTRS)

    Bernacki, Bruce E.

    1992-01-01

    All easy-to-implement focus- and track-error detection methods presently used in magneto-optical (MO) disk drives using pre-grooved media suffer from a side effect known as feedthrough. Feedthrough is the unwanted focus error signal (FES) produced when the optical head is seeking a new track, and light refracted from the pre-grooved disk produces an erroneous FES. Some focus and track-error detection methods are more resistant to feedthrough, but tend to be complicated and/or difficult to keep in alignment as a result of environmental insults. The astigmatic focus/push-pull tracking method is an elegant, easy-to-align focus- and track-error detection method. Unfortunately, it is also highly susceptible to feedthrough when astigmatism is present, with the worst effects caused by astigmatism oriented such that the tangential and sagittal foci are at 45 deg to the track direction. This disclosure outlines a method to nearly completely eliminate the worst-case form of feedthrough due to astigmatism oriented 45 deg to the track direction. Feedthrough due to other primary aberrations is not improved, but performance is identical to the unimproved astigmatic method.

  6. 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.

  7. 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. PMID:26706801

  8. 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.

  9. Robert Feulgen Prize Lecture 1995. New approaches to in situ detection of nucleic acids.

    PubMed

    Thiry, M

    1995-08-01

    The present paper reviews recent results obtained by different molecular biology-based, immunocytological approaches to the localization and identification of nucleic acids in sections of biological material. Examples of sensitive, high-resolution detection methods for RNA, DNA or specialized DNA regions are presented. Special emphasis is placed on the potential values and limitations of these new methods.

  10. Robert Feulgen Prize Lecture 1995. New approaches to in situ detection of nucleic acids.

    PubMed

    Thiry, M

    1995-08-01

    The present paper reviews recent results obtained by different molecular biology-based, immunocytological approaches to the localization and identification of nucleic acids in sections of biological material. Examples of sensitive, high-resolution detection methods for RNA, DNA or specialized DNA regions are presented. Special emphasis is placed on the potential values and limitations of these new methods. PMID:8536076

  11. 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

  12. Intrusion detection using pattern recognition methods

    NASA Astrophysics Data System (ADS)

    Jiang, Nan; Yu, Li

    2007-09-01

    Today, cyber attacks such as worms, scanning, active attackers are pervasive in Internet. A number of security approaches are proposed to address this problem, among which the intrusion detection system (IDS) appears to be one of the major and most effective solutions for defending against malicious users. Essentially, intrusion detection problem can be generalized as a classification problem, whose goal is to distinguish normal behaviors and anomalies. There are many well-known pattern recognition algorithms for classification purpose. In this paper we describe the details of applying pattern recognition methods to the intrusion detection research field. Experimenting on the KDDCUP 99 data set, we first use information gain metric to reduce the dimensionality of the original feature space. Two supervised methods, the support vector machine as well as the multi-layer neural network have been tested and the results display high detection rate and low false alarm rate, which is promising for real world applications. In addition, three unsupervised methods, Single-Linkage, K-Means, and CLIQUE, are also implemented and evaluated in the paper. The low computational complexity reveals their application in initial data reduction process.

  13. 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.

  14. 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.

  15. HPLC-methods for determination of lipoic acid and its reduced form in human plasma.

    PubMed

    Teichert, J; Preiss, R

    1992-11-01

    A method has been developed for the HPLC analysis of lipoic acid and its reduced form (dihydrolipoic acid) in biological samples. Both substances are released from the samples by enzymatic hydrolysis and extracted by solid phase column. The extracts, after evaporation, were chromatographed and quantified by electrochemical detection. The basic level was in the range 1-25 ng/ml for lipoic acid and 33-145 ng/ml for dihydrolipoic acid (6 healthy volunteers).

  16. 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.

  17. [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.

  18. 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.

  19. Graphdiyne as a promising material for detecting amino acids

    PubMed Central

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

    2015-01-01

    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. PMID:26568200

  20. 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.

  1. 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.

  2. Lifting wavelet method of target detection

    NASA Astrophysics Data System (ADS)

    Han, Jun; Zhang, Chi; Jiang, Xu; Wang, Fang; Zhang, Jin

    2009-11-01

    Image target recognition plays a very important role in the areas of scientific exploration, aeronautics and space-to-ground observation, photography and topographic mapping. Complex environment of the image noise, fuzzy, all kinds of interference has always been to affect the stability of recognition algorithm. In this paper, the existence of target detection in real-time, accuracy problems, as well as anti-interference ability, using lifting wavelet image target detection methods. First of all, the use of histogram equalization, the goal difference method to obtain the region, on the basis of adaptive threshold and mathematical morphology operations to deal with the elimination of the background error. Secondly, the use of multi-channel wavelet filter wavelet transform of the original image de-noising and enhancement, to overcome the general algorithm of the noise caused by the sensitive issue of reducing the rate of miscarriage of justice will be the multi-resolution characteristics of wavelet and promotion of the framework can be designed directly in the benefits of space-time region used in target detection, feature extraction of targets. The experimental results show that the design of lifting wavelet has solved the movement of the target due to the complexity of the context of the difficulties caused by testing, which can effectively suppress noise, and improve the efficiency and speed of detection.

  3. Chemical approaches to detect and analyze protein sulfenic acids

    PubMed Central

    Furdui, Cristina M.; Poole, Leslie B.

    2013-01-01

    Orchestration of many processes relying on intracellular signal transduction is recognized to require the generation of hydrogen peroxide as a second messenger, yet relatively few molecular details of how this oxidant acts to regulate protein function are currently understood. This review describes emerging chemical tools and approaches that can be applied to study protein oxidation in biological systems, with a particular emphasis on a key player in protein redox regulation, cysteine sulfenic acid. While sulfenic acids (within purified proteins or simple mixtures) are detectable by physical approaches like X-ray crystallography, nuclear magnetic resonance and mass spectrometry, the propensity of these moieties to undergo further modification in complex biological systems has necessitated the development of chemical probes, reporter groups and analytical approaches to allow for their selective detection and quantification. Provided is an overview of techniques that are currently available for the study of sulfenic acids, and some of the biologically meaningful data that have been collected using such approaches. PMID:24105931

  4. Chemical approaches to detect and analyze protein sulfenic acids.

    PubMed

    Furdui, Cristina M; Poole, Leslie B

    2014-01-01

    Orchestration of many processes relying on intracellular signal transduction is recognized to require the generation of hydrogen peroxide as a second messenger, yet relatively few molecular details of how this oxidant acts to regulate protein function are currently understood. This review describes emerging chemical tools and approaches that can be applied to study protein oxidation in biological systems, with a particular emphasis on a key player in protein redox regulation, cysteine sulfenic acid. While sulfenic acids (within purified proteins or simple mixtures) are detectable by physical approaches like X-ray crystallography, nuclear magnetic resonance and mass spectrometry, the propensity of these moieties to undergo further modification in complex biological systems has necessitated the development of chemical probes, reporter groups and analytical approaches to allow for their selective detection and quantification. Provided is an overview of techniques that are currently available for the study of sulfenic acids, and some of the biologically meaningful data that have been collected using such approaches.

  5. 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

  6. Detecting data anomalies methods in distributed systems

    NASA Astrophysics Data System (ADS)

    Mosiej, Lukasz

    2009-06-01

    Distributed systems became most popular systems in big companies. Nowadays many telecommunications companies want to hold large volumes of data about all customers. Obviously, those data cannot be stored in single database because of many technical difficulties, such as data access efficiency, security reasons, etc. On the other hand there is no need to hold all data in one place, because companies already have dedicated systems to perform specific tasks. In the distributed systems there is a redundancy of data and each system holds only interesting data in appropriate form. Data updated in one system should be also updated in the rest of systems, which hold that data. There are technical problems to update those data in all systems in transactional way. This article is about data anomalies in distributed systems. Avail data anomalies detection methods are shown. Furthermore, a new initial concept of new data anomalies detection methods is described on the last section.

  7. 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.

  8. A molecular beacon, bead-based assay for the detection of nucleic acids by flow cytometry.

    PubMed

    Horejsh, Douglas; Martini, Federico; Poccia, Fabrizio; Ippolito, Giuseppe; Di Caro, Antonino; Capobianchi, Maria R

    2005-01-01

    Molecular beacons are dual-labelled probes that are typically used in real-time PCR assays, but have also been conjugated with solid matrices for use in microarrays or biosensors. We have developed a fluid array system using microsphere-conjugated molecular beacons and the flow cytometer for the specific, multiplexed detection of unlabelled nucleic acids in solution. For this array system, molecular beacons were conjugated with microspheres using a biotin-streptavidin linkage. A bridged conjugation method using streptavidin increased the signal-to-noise ratio, allowing for further discrimination of target quantitation. Using beads of different sizes and molecular beacons in two fluorophore colours, synthetic nucleic acid control sequences were specifically detected for three respiratory pathogens, including the SARS coronavirus in proof-of-concept experiments. Considering that routine flow cytometers are able to detect up to four fluorescent channels, this novel assay may allow for the specific multiplex detection of a nucleic acid panel in a single tube.

  9. 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.

  10. 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.

  11. 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

  12. Analytical method for determination of benzene-arsenic acids

    SciTech Connect

    Mitchell, G.L.; Bayse, G.S.

    1988-01-01

    A sensitive analytical method has been modified for use in determination of several benzenearsonic acids, including arsanilic acid (p-aminobenzenearsonic acid), Roxarsone (3-nitro-4-hydroxybenzenearsonic acid), and p-ureidobenzene arsonic acid. Controlled acid hydrolysis of these compounds produces a quantitative yield of arsenate, which is measured colorimetrically as the molybdenum blue complex at 865 nm. The method obeys Beer's Law over the micromolar concentration range. These benzenearsonic acids are routinely used as feed additives in poultry and swine. This method should be useful in assessing tissue levels of the arsenicals in appropriate extracts.

  13. 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. PMID:27336160

  14. 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

  15. 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).

  16. 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).

  17. 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.

  18. A new IQ detection method for LLRF

    NASA Astrophysics Data System (ADS)

    Qiu, Feng; Gao, Jie; Lin, Hai-ying; Liu, Rong; Ma, Xin-peng; Sha, Peng; Sun, Yi; Wang, Guang-wei; Wang, Qun-yao; Xu, Bo; Zeng, Ri-hua

    2012-05-01

    Digital LLRF technology has been widely used in new generation particle accelerators. IF quadrature sampling is a common method for amplitude and phase detection. Many strategies, which obey the same rule of fsample=(M/N)fIF (M/N is a rational number), have been proposed to reduce the effects of spectrum aliasing. However, we found that M/N does not need to be a rational number according to Shannon's theorem. Therefore, we propose a new IQ detection method in this paper. This method is based on a special IIR filter which is derived from an RLC circuit. The unique characteristic of the method is that the value of fIF is independent of the value of fsample. We have set up an experimental platform to verify our method. A 122.88 MHz sampling clock is used to sample a 3 MHz IF signal. The DDS and PI control techniques are used to realize the closed-loop control. Results show that the stability of the system is within ± 0.05% (peak to peak) for the amplitude, and with ±0.03° (peak to peak) for the phase in 5 h.

  19. 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.

  20. Fluorogenic, catalytic, photochemical reaction for amplified detection of nucleic acids.

    PubMed

    Dutta, Subrata; Fülöp, Annabelle; Mokhir, Andriy

    2013-09-18

    Photochemical, nucleic acid-induced reactions, which are controlled by nontoxic red light, are well-suited for detection of nucleic acids in live cells, since they do not require any additives and can be spatially and temporally regulated. We have recently described the first reaction of this type, in which a phenylselenyl derivative of thymidine (5'-PhSeT-ODNa) is cleaved in the presence of singlet oxygen (Fülöp, A., Peng, X., Greenberg, M. M., Mokhir, A. (2010) A nucleic acid directed, red light-induced chemical reaction. Chem. Commun. 46, 5659-5661). The latter reagent is produced upon exposure of a photosensitizer 3'-PS-ODNb (PS = Indium(III)-pyropheophorbide-a-chloride: InPPa) to >630 nm light. In 2012 we reported on a fluorogenic version of this reaction (Dutta, S., Flottmann, B., Heilemann, M., Mokhir, A. (2012) Hybridization and reaction-based, fluorogenic nucleic acid probes. Chem. Commun. 47, 9664-9666), which is potentially applicable for the detection of nucleic acids in cells. Unfortunately, its yield does not exceed 25% and no catalytic turnover could be observed in the presence of substrate excess. This problem occurs due to the efficient, competing oxidation of the substrate containing an electron rich carbon-carbon double bonds (SCH═CHS) in the presence of singlet oxygen with formation of a noncleavable product (SCH═CHSO). Herein we describe a related, but substantially improved photochemical, catalytic transformation of a fluorogenic, organic substrate, which consists of 9,10-dialkoxyanthracene linked to fluorescein, with formation of a bright fluorescent dye. In highly dilute solution this reaction occurs only in the presence of a nucleic acid template. We developed three types of such a reaction and demonstrated that they are high yielding and generate over 7.7 catalytic turnovers, are sensitive to single mismatches in nucleic acid targets, and can be applied for determination of both the amount of nucleic acids and potentially their

  1. 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.

  2. 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

  3. Waterborne pathogens: detection methods and challenges.

    PubMed

    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

  4. Waterborne pathogens: detection methods and challenges.

    PubMed

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

    2015-05-21

    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.

  5. 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

  6. Sensing Methods for Detecting Analog Television Signals

    NASA Astrophysics Data System (ADS)

    Rahman, Mohammad Azizur; Song, Chunyi; Harada, Hiroshi

    This paper introduces a unified method of spectrum sensing for all existing analog television (TV) signals including NTSC, PAL and SECAM. We propose a correlation based method (CBM) with a single reference signal for sensing any analog TV signals. In addition we also propose an improved energy detection method. The CBM approach has been implemented in a hardware prototype specially designed for participating in Singapore TV white space (WS) test trial conducted by Infocomm Development Authority (IDA) of the Singapore government. Analytical and simulation results of the CBM method will be presented in the paper, as well as hardware testing results for sensing various analog TV signals. Both AWGN and fading channels will be considered. It is shown that the theoretical results closely match with those from simulations. Sensing performance of the hardware prototype will also be presented in fading environment by using a fading simulator. We present performance of the proposed techniques in terms of probability of false alarm, probability of detection, sensing time etc. We also present a comparative study of the various techniques.

  7. Novelty detection using level set methods.

    PubMed

    Ding, Xuemei; Li, Yuhua; Belatreche, Ammar; Maguire, Liam P

    2015-03-01

    This paper presents a level set boundary description (LSBD) approach for novelty detection that treats the nonlinear boundary directly in the input space. The proposed approach consists of level set function (LSF) construction, boundary evolution, and termination of the training process. It employs kernel density estimation to construct the LSF of the initial boundary for the training data set. Then, a sign of the LSF-based algorithm is proposed to evolve the boundary and make it fit more tightly in the data distribution. The training process terminates when an expected fraction of rejected normal data is reached. The evolution process utilizes the signs of the LSF values at all training data points to decide whether to expand or shrink the boundary. Extensive experiments are conducted on benchmark data sets to evaluate the proposed LSBD method and compare it against four representative novelty detection methods. The experimental results demonstrate that the novelty detector modeled with the proposed LSBD can effectively detect anomalies. PMID:25720011

  8. 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)

  9. 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.

  10. 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

    2015-12-01

    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.

  11. 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

    2016-06-01

    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. PMID:27170090

  12. 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

    2016-06-01

    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.

  13. 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.

  14. 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.

  15. 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.

  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. 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

  18. 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.

  19. Dielectric detection by an electromagnetic cavity method

    NASA Astrophysics Data System (ADS)

    Weatherall, James C.; Beckley, Howard F.; Gatto, Joseph A.

    2004-09-01

    A method to screen for flammable and explosive materials in bottles by electromagnetic measurement is described. The technique makes use of an aluminum cavity having strong electromagnetic resonances in the radio wave band. An object inserted into the cavity changes the internal field configuration, and causes small, but measurable shifts in the resonant frequencies. The response depends on the electrical permittivity, or dielectric constant, of the material. By measuring the frequency change of many different cavity modes, the dielectric constant of the perturbing object can be inferred independently of the object's size and shape. We demonstrate the detection of materials in bottles using an experimental device.

  20. 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.

  1. Detection of Acetic Acid in wine by means of an electronic nose

    NASA Astrophysics Data System (ADS)

    Lozano, Jesús; Álvarez, Fernando; Santos, José Pedro; Horrillo, Carmen

    2011-09-01

    A portable electronic nose (see Fig.1) based on metal oxide semiconductor thin-film sensors has been developed to detect acetic acid present in four types of wines. The wines analyzed are from the same cellar but are made with different varieties of grapes. Data analysis was performed by two pattern recognition methods: principal component analysis (PCA) and Artificial Neural Networks (ANN).

  2. 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.

  3. 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

  4. Leak and Pipe Detection Method and System

    NASA Technical Reports Server (NTRS)

    Youngquist, Robert C. (Inventor)

    2003-01-01

    A method and system for locating leaks of conductive fluids from non-conductive pipes and other structures or for locat- ing non-conductive pipes or structures having conductive fluid contained therein, employ a charge generator to apply a time varying charge to the conductive fluid, and a capaci- tive type detector that can detect the variable charge that is induced in the fluid. The capacitive detector, which prefer- ably includes a handheld housing, employs a large conduc- tive pickup plate that is used to locate the pipe or leak by scanning the plate over the ground and detecting the induced charge that is generated when the plate comes in close proximity to the pipe or leak. If a leak is encountered, the resulting signal will appear over an area larger than expected for a buried pipe, assuming the leak provides an electrically conductive path between the flow and the wet surrounding ground. The detector uses any suitable type of indicator device, such as a pair of headphones that enable an operator to hear the detected signal as a chirping sound, for example.

  5. Methods and Instruments for Fast Neutron Detection

    SciTech Connect

    Jordan, David V.; Reeder, Paul L.; Cooper, Matthew W.; McCormick, Kathleen R.; Peurrung, Anthony J.; Warren, Glen A.

    2005-05-01

    Pacific Northwest National Laboratory evaluated the performance of a large-area (~0.7 m2) plastic scintillator time-of-flight (TOF) sensor for direct detection of fast neutrons. This type of sensor is a readily area-scalable technology that provides broad-area geometrical coverage at a reasonably low cost. It can yield intrinsic detection efficiencies that compare favorably with moderator-based detection methods. The timing resolution achievable should permit substantially more precise time windowing of return neutron flux than would otherwise be possible with moderated detectors. The energy-deposition threshold imposed on each scintillator contributing to the event-definition trigger in a TOF system can be set to blind the sensor to direct emission from the neutron generator. The primary technical challenge addressed in the project was to understand the capabilities of a neutron TOF sensor in the limit of large scintillator area and small scintillator separation, a size regime in which the neutral particle’s flight path between the two scintillators is not tightly constrained.

  6. 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.

  7. 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.

  8. 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.

  9. 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. PMID:27479046

  10. Lagrangian based methods for coherent structure detection.

    PubMed

    Allshouse, Michael R; Peacock, Thomas

    2015-09-01

    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. PMID:26428570

  11. Comparison of detection methods for vaginal lactobacilli.

    PubMed

    Smidt, I; Kiiker, R; Oopkaup, H; Lapp, E; Rööp, T; Truusalu, K; Štšepetova, J; Truu, J; Mändar, R

    2015-01-01

    Vaginal lactobacilli offer protection against microbiota imbalance and genitourinary tract infections. We compared vaginal lactobacilli in 50 Estonian women of child-bearing age applying culture-based methods, quantitative PCR and next-generation sequencing (NGS). The culture-based methods found three different lactobacilli: Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus gasseri. Using NGS revealed the presence of L. crispatus in 76%, Lactobacillus iners in 52%, L. jensenii in 47% and L. gasseri in 33% of the samples. According to qPCR, L. iners was present in 67% and L. crispatus in 64% of the samples. The proportions of L. crispatus revealed by qPCR and NGS were in good correlation (R=0.79, P<0.001), while that of L. iners correlated poorly (R=0.13, P>0.05). Good concordance for L. crispatus was also found between the results of the culture-based method and qPCR. Finally, good overlap between the results of the culture-based method and NGS was revealed: in case of a positive NGS result for L. crispatus, the same species was isolated in 95% of samples. The corresponding percentages were 82% for L. jensenii and 86% for L. gasseri. Our data indicate fairly general concordance of the three methods for detecting vaginal lactobacilli, except for L. iners. This points out the importance of standardisation of techniques, and the respective studies should involve cultures applying a medium suitable for the fastidious L. iners. PMID:25869280

  12. 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.

  13. Comparison of detection methods for vaginal lactobacilli.

    PubMed

    Smidt, I; Kiiker, R; Oopkaup, H; Lapp, E; Rööp, T; Truusalu, K; Štšepetova, J; Truu, J; Mändar, R

    2015-01-01

    Vaginal lactobacilli offer protection against microbiota imbalance and genitourinary tract infections. We compared vaginal lactobacilli in 50 Estonian women of child-bearing age applying culture-based methods, quantitative PCR and next-generation sequencing (NGS). The culture-based methods found three different lactobacilli: Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus gasseri. Using NGS revealed the presence of L. crispatus in 76%, Lactobacillus iners in 52%, L. jensenii in 47% and L. gasseri in 33% of the samples. According to qPCR, L. iners was present in 67% and L. crispatus in 64% of the samples. The proportions of L. crispatus revealed by qPCR and NGS were in good correlation (R=0.79, P<0.001), while that of L. iners correlated poorly (R=0.13, P>0.05). Good concordance for L. crispatus was also found between the results of the culture-based method and qPCR. Finally, good overlap between the results of the culture-based method and NGS was revealed: in case of a positive NGS result for L. crispatus, the same species was isolated in 95% of samples. The corresponding percentages were 82% for L. jensenii and 86% for L. gasseri. Our data indicate fairly general concordance of the three methods for detecting vaginal lactobacilli, except for L. iners. This points out the importance of standardisation of techniques, and the respective studies should involve cultures applying a medium suitable for the fastidious L. iners.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

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

    SciTech Connect

    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.

  19. NAIL: Nucleic Acid detection using Isotachophoresis and Loop-mediated isothermal amplification.

    PubMed

    Borysiak, Mark D; Kimura, Kevin W; Posner, Jonathan D

    2015-04-01

    Nucleic acid amplification tests are the gold standard for many infectious disease diagnoses due to high sensitivity and specificity, rapid operation, and low limits of detection. Despite the advantages of nucleic acid amplification tests, they currently offer limited point-of-care (POC) utility due to the need for complex instruments and laborious sample preparation. We report the development of the Nucleic Acid Isotachophoresis LAMP (NAIL) diagnostic device. NAIL uses isotachophoresis (ITP) and loop-mediated isothermal amplification (LAMP) to extract and amplify nucleic acids from complex matrices in less than one hour inside of an integrated chip. ITP is an electrokinetic separation technique that uses an electric field and two buffers to extract and purify nucleic acids in a single step. LAMP amplifies nucleic acids at constant temperature and produces large amounts of DNA that can be easily detected. A mobile phone images the amplification results to eliminate the need for laser fluorescent detection. The device requires minimal user intervention because capillary valves and heated air chambers act as passive valves and pumps for automated fluid actuation. In this paper, we describe NAIL device design and operation, and demonstrate the extraction and detection of pathogenic E. coli O157:H7 cells from whole milk samples. We use the Clinical and Laboratory Standards Institute (CLSI) limit of detection (LoD) definitions that take into account the variance from both positive and negative samples to determine the diagnostic LoD. According to the CLSI definition, the NAIL device has a limit of detection (LoD) of 1000 CFU mL(-1) for E. coli cells artificially inoculated into whole milk, which is two orders of magnitude improvement to standard tube-LAMP reactions with diluted milk samples and comparable to lab-based methods. The NAIL device potentially offers significant reductions in the complexity and cost of traditional nucleic acid diagnostics for POC applications

  20. NAIL: Nucleic Acid detection using Isotachophoresis and Loop-mediated isothermal amplification.

    PubMed

    Borysiak, Mark D; Kimura, Kevin W; Posner, Jonathan D

    2015-04-01

    Nucleic acid amplification tests are the gold standard for many infectious disease diagnoses due to high sensitivity and specificity, rapid operation, and low limits of detection. Despite the advantages of nucleic acid amplification tests, they currently offer limited point-of-care (POC) utility due to the need for complex instruments and laborious sample preparation. We report the development of the Nucleic Acid Isotachophoresis LAMP (NAIL) diagnostic device. NAIL uses isotachophoresis (ITP) and loop-mediated isothermal amplification (LAMP) to extract and amplify nucleic acids from complex matrices in less than one hour inside of an integrated chip. ITP is an electrokinetic separation technique that uses an electric field and two buffers to extract and purify nucleic acids in a single step. LAMP amplifies nucleic acids at constant temperature and produces large amounts of DNA that can be easily detected. A mobile phone images the amplification results to eliminate the need for laser fluorescent detection. The device requires minimal user intervention because capillary valves and heated air chambers act as passive valves and pumps for automated fluid actuation. In this paper, we describe NAIL device design and operation, and demonstrate the extraction and detection of pathogenic E. coli O157:H7 cells from whole milk samples. We use the Clinical and Laboratory Standards Institute (CLSI) limit of detection (LoD) definitions that take into account the variance from both positive and negative samples to determine the diagnostic LoD. According to the CLSI definition, the NAIL device has a limit of detection (LoD) of 1000 CFU mL(-1) for E. coli cells artificially inoculated into whole milk, which is two orders of magnitude improvement to standard tube-LAMP reactions with diluted milk samples and comparable to lab-based methods. The NAIL device potentially offers significant reductions in the complexity and cost of traditional nucleic acid diagnostics for POC applications.

  1. [Detection of cyclopiazonic acid and its producers in food].

    PubMed

    Ostrý, V; Polster, M

    1989-07-01

    In the course of six months, 60 samples of foods were examined for their contents of cyclopiazonic acid. These samples were subjected to a basal mycological screening aimed at Aspergillus flavus and Penicillium sp. strains. Cyclopiazonic acid contents in samples of Hermelín cheese, peanuts, rice, peeled barley grains, Folican salami, and packaged meat did not exceed the value of 0.5 mg.kg-1. When using a modification of the method of cyclopiazonic acid isolation described by Dorner et al. (1983), 521 mg of this mycotoxin were isolated from a culture of Penicillium griseofulvum CCM 8006 strain grown in liquid medium containing 2% yeast autolysate and 2.5% sucrose. About 47% of the isolated Aspergillus flavus strains were bitoxicogenic (produced both cyclopiazonic acid and aflatoxin). Cyclopiazonic acid was produced by 23.5% of the isolated Penicillium sp. strains. No cyclopiazonic acid was produced in vitro by Penicillium nalgoviensis strains from the Czechoslovak collection on sweet wort agar containing peptone from soybean. Penicillium commune F-426 and Penicillium aurantiogriseum F-708 strains are efficient producers of this acid. PMID:2508296

  2. [Detection of cyclopiazonic acid and its producers in food].

    PubMed

    Ostrý, V; Polster, M

    1989-07-01

    In the course of six months, 60 samples of foods were examined for their contents of cyclopiazonic acid. These samples were subjected to a basal mycological screening aimed at Aspergillus flavus and Penicillium sp. strains. Cyclopiazonic acid contents in samples of Hermelín cheese, peanuts, rice, peeled barley grains, Folican salami, and packaged meat did not exceed the value of 0.5 mg.kg-1. When using a modification of the method of cyclopiazonic acid isolation described by Dorner et al. (1983), 521 mg of this mycotoxin were isolated from a culture of Penicillium griseofulvum CCM 8006 strain grown in liquid medium containing 2% yeast autolysate and 2.5% sucrose. About 47% of the isolated Aspergillus flavus strains were bitoxicogenic (produced both cyclopiazonic acid and aflatoxin). Cyclopiazonic acid was produced by 23.5% of the isolated Penicillium sp. strains. No cyclopiazonic acid was produced in vitro by Penicillium nalgoviensis strains from the Czechoslovak collection on sweet wort agar containing peptone from soybean. Penicillium commune F-426 and Penicillium aurantiogriseum F-708 strains are efficient producers of this acid.

  3. Analysis of neurotransmitter amino acids by CE-LIF detection in biological fluids.

    PubMed

    Zinellu, Angelo; Sotgia, Salvatore; Deiana, Luca; Carru, Ciriaco

    2013-01-01

    Neurotransmitter amino acids like Alanine (Ala), Glutamic acid (Glu), Aspartic acid (Asp), Serine (Ser), Taurine (Tau), and Glycine (Gly) are widely distributed in biological fluids and tissues and they have important physiological functions, so that the evaluation of their levels in organisms plays an essential role in physiology. We here report a new capillary electrophoresis (CE) method with laser-induced fluorescence detection by which analytes are resolved and detected in less than 12 min both in human plasma and in other samples types, such as red blood cells, urine, cultured cells, cerebrospinal fluid, saliva, and vitreous humor, thus avoiding the typical limitations of the other methods normally suitable only in one or two matrix types.

  4. 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.

  5. Efficient Computer Network Anomaly Detection by Changepoint Detection Methods

    NASA Astrophysics Data System (ADS)

    Tartakovsky, Alexander G.; Polunchenko, Aleksey S.; Sokolov, Grigory

    2013-02-01

    We consider the problem of efficient on-line anomaly detection in computer network traffic. The problem is approached statistically, as that of sequential (quickest) changepoint detection. A multi-cyclic setting of quickest change detection is a natural fit for this problem. We propose a novel score-based multi-cyclic detection algorithm. The algorithm is based on the so-called Shiryaev-Roberts procedure. This procedure is as easy to employ in practice and as computationally inexpensive as the popular Cumulative Sum chart and the Exponentially Weighted Moving Average scheme. The likelihood ratio based Shiryaev-Roberts procedure has appealing optimality properties, particularly it is exactly optimal in a multi-cyclic setting geared to detect a change occurring at a far time horizon. It is therefore expected that an intrusion detection algorithm based on the Shiryaev-Roberts procedure will perform better than other detection schemes. This is confirmed experimentally for real traces. We also discuss the possibility of complementing our anomaly detection algorithm with a spectral-signature intrusion detection system with false alarm filtering and true attack confirmation capability, so as to obtain a synergistic system.

  6. Luminescent Probes for Ultrasensitive Detection of Nucleic Acids

    PubMed Central

    Krasnoperov, Lev N.; Marras, Salvatore A.E.; Kozlov, Maxim; Wirpsza, Laura; Mustaev, Arkady

    2010-01-01

    Novel amino-reactive derivatives of lanthanide-based luminescent labels of enhanced brightness and metal retention were synthesized and used for the detection of complementary DNA oligonucleotides by molecular beacons. Time-resolved acquisition of the luminescent signal that occurs upon hybridization of the probe to the target enabled the avoidance of short-lived background fluorescence, markedly enhancing the sensitivity of detection, which was less than 1 pM. This value is about 50 to 100 times more sensitive than the level achieved with conventional fluorescence-based molecular beacons, and is 10 to 60 times more sensitive than previously reported for other lanthanide-based hybridization probes. These novel luminescent labels should significantly enhance the sensitivity of all type of nucleic acid hybridization probes, and could dramatically improve the detection limit of other biopolymers and small compounds that are used in a variety of biological applications. PMID:20085336

  7. 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

  8. 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).

  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. Collection and analysis of organic acids in exhaust gas. Comparison of different methods

    NASA Astrophysics Data System (ADS)

    Zervas, E.; Montagne, X.; Lahaye, J.

    This paper reports the development of a specific method to identify organic acids in exhaust gases. The organic acids are collected in two impingers containing liquids (pure water or Na 2CO 3 1% aqueous solution) and four cartridges containing solids (silica, fluorisil, alumina B and alumina N). Once collected, the acids are eluted of the solids by a hot water stream. These traps performances, in terms of organic acids collection and elution efficiency, are evaluated and compared. Two sources are used to produce the gas flow containing organic acids: one generates a flow whose concentration is known and stable, the other produces organic acids among other combustion products. For eluted solutions analysis, two methods are used: isocratic ionic chromatography/conductivity detection and GC/FID. Their efficiency in separating 10 aliphatic acids are compared. Their characteristics such as detection limits, detection linearity, repeatability and possible interferences with other components found in exhaust gases are determined. The stability of the organic acids solutions is also studied. Lastly, the use of these methods is illustrated by the analysis of the gas-phase organic acids exhausted by a spark ignition and by a diesel engine.

  11. 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.

  12. Liquid chromatography detection unit, system, and method

    SciTech Connect

    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.

  13. 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.

  14. 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.

  15. Electrophysiological methods in early glaucoma detection.

    PubMed

    Fernández-Tirado, F J; Uclés, P; Pablo, L; Honrubia, F M

    1994-04-01

    Electrophysiological methods like pattern electroretinogram and pattern visual evoked potentials may be the best approach for early glaucoma detection, as they have demonstrated to be sensitive to the ganglion cells functional changes. Reported values of both P50 and N95 pattern electroretinogram components differ widely, a fact perhaps related to a heterogeneous grouping of patients suffering from ocular hypertension and glaucoma, rather than to the type of technique utilized. This study is based on a total of 42 subjects: 14 normals, 16 subjects suffering from ocular hypertension, and 12 patients with glaucoma, with 79 eyes examined. Pattern electroretinogram and pattern visual evoked potentials were used as successive techniques. Setting condition for pattern electroretinogram, such as low temporal frequency (2 Hz), 30 min check size, high contrast (99%) and luminance (93 cd/m2), identified the N95 component as the best index for early glaucoma detection. In the control group N95 mean amplitude was 1.62 muV +/- 0.59 SD and showed almost significant difference with ocular hypertension (p = 0.07) and highly significant difference with the glaucoma group (p < 0.01), with decrement of 58.6% in the glaucoma group. P50 mean amplitude, on the contrary, did not show significant differences among the groups (Newman-Keul test), its reduction in glaucoma being 28%. The mean pattern visual evoked potentials latency was alos highly significant between glaucoma and control groups, but not between ocular hypertension and control groups; the mean amplitude did not show significant differences. A proportion of 26.6% abnormal pattern electroretinogram was found in the group suspected of having glaucoma where conventional methods had proved normal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8079620

  16. 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

  17. 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.

  18. Detection of COL III in parchment by amino acid analysis.

    PubMed

    Sommer, Dorte V P; Larsen, René

    2016-01-01

    Cultural heritage parchments made from the reticular dermis of animals have been subject to studies of deterioration and conservation by amino acid analysis. The reticular dermis contains a varying mixture of collagen I and III (COL I and III). When dealing with the results of the amino acid analyses, till now the COL III content has not been taken into account. Based on the available amino acid sequences, we present a method for determining the amount of COL III in the reticular dermis of new and historical parchments calculated from the ratio of Ile/Val. We find COL III contents between 7 and 32 % in new parchments and between 0.2 and 40 % in the historical parchments. This is consistent with results in the literature. The varying content of COL III has a significant influence on the uncertainty of the amino acid analysis. Although we have not found a simple correlation between the COL III content and the degree of deterioration, our results show that this question must be taken into consideration in future studies of the chemical and physical deterioration of parchment measured by amino acid analysis and other analytical methods.

  19. [Determination of organic acids in cane vinasse by micellar electrokinetic capillary chromatography with indirect ultraviolet detection].

    PubMed

    Xu, Yuanjin; Xu, Guiping; Wei, Yuanan

    2006-01-01

    Micellar electrokinetic capillary chromatography (MECC) with indirect ultraviolet (UV) detection method for the separation and determination of several organic acids in cane vinasse, including malonic, formic, tartaric, malic, succinic, glutaric, acetic, lactic and glutamic acids, were developed. Electrophoretic conditions were as follows: uncoated fused silica capillary (56 cm/ 64 cm (effective/total length), 50 microm i. d. ), 7.5 mmol/L potassium acid phthalate-1. 5 mmol/L cetyltrimethyl-ammonium bromide (CTAB) at pH = 6.50 as buffer solution, applied voltage -25 kV, temperature 25 degrees C, detection wavelength 300 nm, reference wavelength 210 nm. Good linearities were obtained for nine organic acids, and the detection limits were 0.5 mg/L, 0.3 mg/L, 1.5 mg/L, 1.5 mg/L, 0.3 mg/L, 0.3 mg/L, 0.4 mg/L, 0.4 mg/L, 0.4 mg/L for malonic, formic, tartaric, malic, succinic, glutaric, acetic, lactic and glutamic acid, respectively. The relative standard deviations (RSDs) for migration times and peak areas of nine organic acids within a day were 0.4% - 0.6% and 2.3% - 4.8%, respectively. The corresponding data for five days were 0.5% -0.7% and 3.3% - 5.2%. The recoveries of acid standards were above 93%. The method can be applied to determine the organic acids in cane vinasse with satisfactory results. PMID:16827307

  20. 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

  1. DETECTION OF A NEW INTERSTELLAR MOLECULE: THIOCYANIC ACID HSCN

    SciTech Connect

    Halfen, D. T.; Ziurys, L. M.; Bruenken, S.; Gottlieb, C. A.; McCarthy, M. C.; Thaddeus, P. E-mail: lziurys@as.arizona.edu E-mail: cgottlieb@cfa.harvard.edu E-mail: pthaddeus@cfa.harvard.edu

    2009-09-10

    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 K{sub a} = 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 x 10{sup 13} cm{sup -2} in Sgr B2(N) is only three times lower than that of HNCS. The fractional abundances of HSCN and HNCS relative to H{sub 2} are 4.5 x 10{sup -12} and 1.1 x 10{sup -11}. By analogy with the isomeric pair HCN and HNC, these two sulfur-bearing isomers are plausibly formed from a common cation precursor.

  2. 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.

  3. Spectrophotometric method for determining gibberellic acid in fermentation broths.

    PubMed

    Berríos, Julio; Illanes, Andrés; Aroca, Germán

    2004-01-01

    A novel method for the quantitative determination of gibberellic acid in fermentation broths has been developed. It is based on the kinetic of the reaction of conversion of gibberellic acid to gibberellenic acid. The method is simple, reliable, faster than most of methods known, and free of the interferences which commonly affect spectrophotometric methods currently in use. Its threshold sensitivity is 0.1 g and its accuracy is greater than 97% for concentrations of gibberellic acid ranging from 0.1 to 1 g l(-1).

  4. Comparison of Rapid Methods for Analysis of Bacterial Fatty Acids

    PubMed Central

    Moss, C. Wayne; Lambert, M. A.; Merwin, W. H.

    1974-01-01

    When rapid gas-liquid chromatography methods for determination of bacterial fatty acids were compared, results showed that saponification was required for total fatty acid analysis. Transesterification with boron-trihalide reagents (BF3-CH3OH, BCl3-CH3OH) caused extensive degradation of cyclopropane acids and was less effective than saponification in releasing cellular hydroxy fatty acids. Digestion of cells with tetramethylammonium hydroxide was unsatisfactory because of extraneous gas-liquid chromatography peaks and because of lower recovery of branched-chain and hydroxy fatty acids. A simple, rapid saponification procedure which can be used for total cellular fatty acid analysis of freshly grown cells is described. PMID:4844271

  5. 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.

  6. Sensitive and selective electrochemical detection of artemisinin based on its reaction with p-aminophenylboronic acid.

    PubMed

    Wang, Chao; Zholudov, Yuriy T; Nsabimana, Anaclet; Xu, Guobao; Li, Jianping

    2016-09-21

    The electrochemical detection of artemisinin generally requires high oxidation potential or the use of complex electrode modification. We find that artemisinin can react with p-aminophenylboronic acid to produce easily electrochemically detectable aminophenol for the first time. By making use of the new reaction, we report an alternative method to detect artemisinin through the determination of p-aminophenol. The calibration curve for the determination of artemisinin is linear in the range of 2 μmol L(-1) to 200 μmol L(-1) with the detection limit of 0.8 μmol L(-1), which is more sensitive than other reported electrochemical methods. The relative standard deviation is 4.83% for the determination of 10 μM artemisinin. Because the oxidation potential of p-aminophenol is around 0 V, the present method is high selective. When 40 μM, 90 μM and 140 μM of artemisinin were spiked to compound naphthoquine phosphate tablet samples, the recoveries are 107.6%, 105.4% and 101.7%, respectively. This detection strategy is attractive for the detection of artemisinin and its derivatives. The finding that artemisinin can react with aromatic boronic acid has the potential to be exploited for the development of other sensors, such as fluorescence artemisinin sensors. PMID:27590543

  7. 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.

  8. 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.

  9. DOM Based XSS Detecting Method Based on Phantomjs

    NASA Astrophysics Data System (ADS)

    Dong, Ri-Zhan; Ling, Jie; Liu, Yi

    Because malicious code does not appear in html source code, DOM based XSS cannot be detected by traditional methods. By analyzing the causes of DOM based XSS, this paper proposes a detection method of DOM based XSS based on phantomjs. This paper uses function hijacking to detect dangerous operation and achieves a prototype system. Comparing with existing tools shows that the system improves the detection rate and the method is effective to detect DOM based XSS.

  10. 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.

  11. Optimization of methods for detecting norovirus on various fruit.

    PubMed

    Kim, Hee-Yeon; Kwak, In-Shin; Hwang, In-Gyun; Ko, GwangPyo

    2008-11-01

    Methods for detecting norovirus (NoV) in food are crucial for investigation and prevention of outbreaks caused by NoV-contaminated food. However, current NoV detection methods have not been well examined or optimized. In this study, the effectiveness of various methods for eluting NoV from various fruit, concentrating the virus using polyethylene glycol (PEG), and extracting the viral RNA for subsequent assay by RT-PCR was optimized. First, six different buffers previously described for eluting NoV from fruit surfaces were evaluated. A known amount of NoV was spiked onto the surface of grapes, strawberries, and raspberries, and the virus was recovered with distilled water, 0.05 M glycine-0.14 M NaCl (pH 7.5), 2.9% tryptose phosphate broth-6% glycine, 100 mM Tris-HCl (pH 9.5), 50 mM glycine-50 mM MgCl(2) (pH 9.5), or 3% beef extract. Quantitation of the recovered virus using RT-PCR revealed that the most effective elution buffer was 3% beef extract. Secondly, to optimize a method for concentrating the recovered NoV, the key parameters of PEG precipitation, a typical method for concentrating enteric virus, were investigated. The influence of PEG molecular weight and the duration and temperature of the precipitation procedure were examined. NoV was concentrated most efficiently by precipitation when PEG (10,000) was used for 4h at room temperature. Finally, five different methods for nucleic acid extraction were evaluated. Among RNA extraction methods examined, QIAamp Viral RNA Mini kit showed the best recovery efficiency. Using the optimized method, approximately 6-80% of the seeded NoV was recovered from the various fruit.

  12. Assessment criteria and approaches for rapid detection methods to be used in the food industry.

    PubMed

    Wiedmann, Martin; Wang, Siyun; Post, Laurie; Nightingale, Kendra

    2014-04-01

    The number of commercially available kits and methods for rapid detection of foodborne pathogens continues to increase at a considerable pace, and the diversity of methods and assay formats is reaching a point where it is very difficult even for experts to weigh the advantages and disadvantages of different methods and to decide which methods to choose for a certain testing need. Although a number of documents outline quantitative criteria that can be used to evaluate different detection methods (e.g., exclusivity and inclusivity), a diversity of criteria is typically used by industry to select specific methods that are used for pathogen detection. This article is intended to provide an overall outline of criteria that the food industry can use to evaluate new rapid detection methods, with a specific focus on nucleic acid-based detection methods.

  13. 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%.

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

    DOEpatents

    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.

  15. 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.

  16. Nanostructure Modified Microelectrode for Electrochemical Detection of Dopamine with Ascorbic Acid and Uric Acid.

    PubMed

    Kim, Kyeong-Jun; Choi, Jin-Ha; Pyo, Su-Hyun; Yun, Kwang-Seok; Lee, Ji-Young; Choi, Jeong-Woo; Oh, Byung-Keun

    2016-03-01

    Dopamine (DA) is one kind of neurotransmitter in central nervous system which is indicator of neural disease. For this reason, determination of DA concentration in central nervous system is very important for early diagnosis of neural disease. In this study, we designed micro electrode array and fabricated by MEMS technology. Furthermore, we fabricated 3-D conducting nanostructure on electrode surface for enhanced sensitivity and selectivity due to increased surface area. Compared with macro and normal micro electrode, the 3-D nanostructure modified micro electrode shows better electrical performance. These surface modified pin type electrode was applied to detect low concentration of DA and successfully detect various concentration of DA from 100 μM to 1 μM with linear relationship in the presence of ascorbic acid and uric acid. From these results, our newly designed electrode shows possibility to be applied as brain biosensor for neural disease diagnosis such as Parkinson's diseases. PMID:27455760

  17. Integrated sample-to-detection chip for nucleic acid test assays.

    PubMed

    Prakash, R; Pabbaraju, K; Wong, S; Tellier, R; Kaler, K V I S

    2016-06-01

    Nucleic acid based diagnostic techniques are routinely used for the detection of infectious agents. Most of these assays rely on nucleic acid extraction platforms for the extraction and purification of nucleic acids and a separate real-time PCR platform for quantitative nucleic acid amplification tests (NATs). Several microfluidic lab on chip (LOC) technologies have been developed, where mechanical and chemical methods are used for the extraction and purification of nucleic acids. Microfluidic technologies have also been effectively utilized for chip based real-time PCR assays. However, there are few examples of microfluidic systems which have successfully integrated these two key processes. In this study, we have implemented an electro-actuation based LOC micro-device that leverages multi-frequency actuation of samples and reagents droplets for chip based nucleic acid extraction and real-time, reverse transcription (RT) PCR (qRT-PCR) amplification from clinical samples. Our prototype micro-device combines chemical lysis with electric field assisted isolation of nucleic acid in a four channel parallel processing scheme. Furthermore, a four channel parallel qRT-PCR amplification and detection assay is integrated to deliver the sample-to-detection NAT chip. The NAT chip combines dielectrophoresis and electrostatic/electrowetting actuation methods with resistive micro-heaters and temperature sensors to perform chip based integrated NATs. The two chip modules have been validated using different panels of clinical samples and their performance compared with standard platforms. This study has established that our integrated NAT chip system has a sensitivity and specificity comparable to that of the standard platforms while providing up to 10 fold reduction in sample/reagent volumes.

  18. 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.

  19. Method for the separation of acid from acid-laden vapors

    SciTech Connect

    Hansen, L.J.

    1992-02-11

    This patent describes a method for the removal of hydrochloric or sulfuric acid from vapor laden with the acid. It comprises: contacting the acid-laden vapors with packing materials in a zone containing the packing materials wherein the packing materials are formed of polyester resin containing from about 5 to 40 weight percent aluminum sulfate crystals.

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. Method for Enzyme Design with Genetically Encoded Unnatural Amino Acids.

    PubMed

    Hu, C; Wang, J

    2016-01-01

    We describe the methodologies for the design of artificial enzymes with genetically encoded unnatural amino acids. Genetically encoded unnatural amino acids offer great promise for constructing artificial enzymes with novel activities. In our studies, the designs of artificial enzyme were divided into two steps. First, we considered the unnatural amino acids and the protein scaffold separately. The scaffold is designed by traditional protein design methods. The unnatural amino acids are inspired by natural structure and organic chemistry methods, and synthesized by either organic chemistry methods or enzymatic conversion. With the increasing number of published unnatural amino acids with various functions, we described an unnatural amino acids toolkit containing metal chelators, redox mediators, and click chemistry reagents. These efforts enable a researcher to search the toolkit for appropriate unnatural amino acids for the study, rather than design and synthesize the unnatural amino acids from the beginning. After the first step, the model enzyme was optimized by computational methods and directed evolution. Lastly, we describe a general method for evolving aminoacyl-tRNA synthetase and expressing unnatural amino acids incorporated into a protein. PMID:27586330

  5. Method for Enzyme Design with Genetically Encoded Unnatural Amino Acids.

    PubMed

    Hu, C; Wang, J

    2016-01-01

    We describe the methodologies for the design of artificial enzymes with genetically encoded unnatural amino acids. Genetically encoded unnatural amino acids offer great promise for constructing artificial enzymes with novel activities. In our studies, the designs of artificial enzyme were divided into two steps. First, we considered the unnatural amino acids and the protein scaffold separately. The scaffold is designed by traditional protein design methods. The unnatural amino acids are inspired by natural structure and organic chemistry methods, and synthesized by either organic chemistry methods or enzymatic conversion. With the increasing number of published unnatural amino acids with various functions, we described an unnatural amino acids toolkit containing metal chelators, redox mediators, and click chemistry reagents. These efforts enable a researcher to search the toolkit for appropriate unnatural amino acids for the study, rather than design and synthesize the unnatural amino acids from the beginning. After the first step, the model enzyme was optimized by computational methods and directed evolution. Lastly, we describe a general method for evolving aminoacyl-tRNA synthetase and expressing unnatural amino acids incorporated into a protein.

  6. 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.

  7. Gallic Acid: Review of the Methods of Determination and Quantification.

    PubMed

    Fernandes, Felipe Hugo Alencar; Salgado, Hérida Regina Nunes

    2016-05-01

    Gallic acid (3,4,5 trihydroxybenzoic acid) is a secondary metabolite present in most plants. This metabolite is known to exhibit a range of bioactivities including antioxidant, antimicrobial, anti-inflammatory, and anticancer. There are various methods to analyze gallic acid including spectrometry, chromatography, and capillary electrophoresis, among others. They have been developed to identify and quantify this active ingredient in most biological matrices. The aim of this article is to review the available information on analytical methods for gallic acid, as well as presenting the advantages and limitations of each technique.

  8. Analysis of the oxocarboxylic acid fraction in serum and urine as O-methyloximes by thermionic specific detection.

    PubMed

    Liebich, H M; Pickert, A; Wöll, J

    1981-11-01

    After O-methyloximation of the carbonyl functions, the organic acids in serum and urine are extracted by anion-exchange chromatography, transformed into the methyl esters and pre-fractionated by thin-layer chromatography. In one of the four fractions and on the basis of the nitrogen in the O-methyloxime esters, the profiles of the oxocarboxylic acids are analyzed by gas chromatography with thermionic specific detection. The method has good specificity for the oxocarboxylic acids and is suitable for comparative studies. During diabetic or fasting ketoacidosis the serum concentrations of 3-oxobutyric acid and of the amino acid metabolites 2-oxobutyric, 2-oxoisovaleric and especially 2-oxo-3-methylvaleric and 2-oxoisocaproic acid are increased. In urine mainly 3-oxobutyric acid and only small amounts of the 2-oxocarboxylic acids are excreted.

  9. Β-hydroxymyristic acid as a chemical marker to detect endotoxins in dialysis water.

    PubMed

    Mishra, Rupesh K; Robert-Peillard, Fabien; Ravier, Sylvain; Coulomb, Bruno; Boudenne, Jean-Luc

    2015-02-01

    An analytical chemical method has been developed for determination of β-hydroxymyristic acid (β-HMA), a component of lipopolysaccharides (LPSs/endotoxins) in dialysis water. In our investigation, the β-HMA component was used as a chemical marker for endotoxin presence in dialysis water because it is available in the molecular subunit (lipid A) and responsible for toxicity. It is the most abundant saturated fatty acid in that subunit. The developed method is based on fluorescence derivatization with 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ). A high-performance liquid chromatographic separation of the β-HMA derivative was achieved using an octadecyl silica column in gradient elution. A wide dynamic range of β-HMA was tested and a calibration curve was constructed with accuracy of 90% and variability of less than 10%. The limits of detection and quantification obtained were 2 and 5μM, respectively. The developed method was applied to detect endotoxins in dialysis water by alkaline hydrolysis of LPS using NaOH (0.25M) at 60°C for 2h. After hydrolysis, free acid was detected as its NBD-PZ derivative using high-performance liquid chromatography/mass spectrometry (HPLC/MS). Good recovery rates ranging from 98 to 105% were obtained for β-HMA in dialysis water.

  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. 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.

  12. 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

    2015-06-01

    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.

  13. [Chromatographic determination of bile acids in biological fluids with sensitive and selective detection].

    PubMed

    Goto, J

    1990-11-01

    Separation and determination of bile acids in biological fluids by high-performance liquid chromatography are reviewed. The capacity ratios of bile acids on an ODS column were affected by the number, position and configuration of the hydroxyl group on the steroid nucleus, and the chromatographic behavior was markedly influenced by the pH of a mobile phase according to the conjugated form at C-24. A new pre-column derivatization reagent, 1-anthroyl nitrile, was developed and applied to the analysis of bile acids in biological fluids. Bile acids were derivatized through the 3 alpha-hydroxyl group into the corresponding esters, separated on an ODS column, and monitored by a fluorescence detector with detection limit of 20 fmol. The sensitive method for the determination of bile acids in biological materials by gas chromatography (GC) in combination with negative ion chemical ionization (NICI) mass spectrometry is also described. Of various derivatives for the carboxyl group, the pentafluorobenzyl (PFB) ester provided the highest value of the ratio of the negative to positive ion current. A characteristic carboxylate anion [M-181]- was produced as the most abundant ion by the loss of the PFB group in NICI. PFB esters of bile acids were further derivatized into the dimethylethylsilyl ethers and then separated by GC. The detection limit was 2 fg when the characteristic anion was monitored in the NICI mode. The preparation of 18O-labelled bile acids, as the internal standard for the trace analysis or the tracer for the metabolic study, was developed. Finally, the clean-up procedure for bile acids in biological fluids was investigated. The combined use of solid-phase extraction with a Sep-pak C18 or Bond Elut cartridge and group separation on a lipophilic ion-exchange gel, piperidinohydroxypropyl Sephadex LH-20, was found most effective for this purpose. PMID:2082011

  14. 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.

  15. Molecular detection of harmful algal blooms (HABs) using locked nucleic acids and bead array technology.

    PubMed

    Diaz, Mara R; Jacobson, James W; Goodwin, Kelly D; Dunbar, Sherry A; Fell, Jack W

    2010-06-01

    Harmful algal blooms (HABs) are a serious public health risk in coastal waters. As the intensity and frequency of HABs continue to rise, new methods of detection are needed for reliable identification. Herein, we developed a high-throughput, multiplex, bead array technique for the detection of the dinoflagellates Karenia brevis and Karenia mikimotoi. The method combined the Luminex detection system with two novel technologies: locked nucleic acid-modified oligonucleotides (LNA) and Mirus Label IT(®) nucleic acid technology. To study the feasibility of the method, we evaluated the performance of modified and unmodified LNA probes with amplicon targets that were biotin labeled with two different strategies: direct chemical labeling (Mirus Label IT) versus enzymatic end-labeling (single biotinylated primer). The results illustrated that LNA probes hybridized to complementary single-stranded DNA with better affinity and displayed higher fluorescence intensities than unmodified oligonucleotide DNA probes. The latter effect was more pronounced when the assay was carried out at temperatures above 53°C degree. As opposed to the enzymatic 5' terminal labeling technique, the chemical-labeling method enhanced the level of fluorescence by as much as ~83%. The detection limits of the assay, which were established with LNA probes and Mirus Label IT system, ranged from 0.05 to 46 copies of rRNA. This high-throughput method, which represents the first molecular detection strategy to integrate Luminex technology with LNA probes and Mirus Label IT, can be adapted for the detection of other HABs and is well suited for the monitoring of red tides at pre-blooming and blooming conditions.

  16. [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.

  17. TiO2 optical sensor for amino acid detection

    NASA Astrophysics Data System (ADS)

    Tereshchenko, Alla; Viter, Roman; Konup, Igor; Ivanitsa, Volodymyr; Geveliuk, Sergey; Ishkov, Yuriy; Smyntyna, Valentyn

    2013-11-01

    A novel optical sensor based on TiO2 nanoparticles for Valine detection has been developed. In the presented work, commercial TiO2 nanoparticles (Sigma Aldrich, particle size 32 nm) were used as sensor templates. The sensitive layer was formed by a porphyrin coating on a TiO2 nanostructured surface. As a result, an amorphous layer between the TiO2 nanostructure and porphyrin was formed. Photoluminescence (PL) spectra were measured in the range of 370-900 nm before and after porphyrin application. Porphyrin adsorption led to a decrease of the main TiO2 peak at 510 nm and the emergence of an additional peak of high intensity at 700 nm. Absorption spectra (optical density vs. wavelenght, measured from 300 to 1100 nm) showed IR shift Sorret band of prophiryn after deposition on metal oxide. Adsorption of amino acid quenched PL emission, related to porphyrin and increased the intensity of the TiO2 emission. The interaction between the sensor surface and the amino acid leads to the formation of new complexes on the surface and results in a reduction of the optical activity of porphyrin. Sensitivity of the sensor to different concentrations of Valine was calculated. The developed sensor can determine the concentration of Valine in the range of 0.04 to 0.16 mg/ml.

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

    PubMed

    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

  19. Real-time nucleic acid sequence-based amplification assay for detection of hepatitis A virus.

    PubMed

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

    2005-11-01

    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.

  20. A new acid-fast trichrome stain for simultaneous detection of Cryptosporidium parvum and microsporidial species in stool specimens.

    PubMed

    Ignatius, R; Lehmann, M; Miksits, K; Regnath, T; Arvand, M; Engelmann, E; Futh, U; Hahn, H; Wagner, J

    1997-02-01

    The detection in stool specimens of Cryptosporidium parvum and microsporidia, the most frequent parasitic pathogens causing diarrhea in AIDS patients, until now has depended on two different staining methods. However, since double infections occur and minimization of laboratory costs is mandatory, development of a method for simultaneous detection of these parasites appeared desirable. We report on a new, inexpensive, and easy-to-perform staining procedure to demonstrate both acid-fast oocysts of C. parvum and other coccidia, as well as microsporidial spores. This acid-fast trichrome stain yields results comparable to those obtained by the Kinyoun and modified trichrome methods and considerably reduces the time necessary for microscopic examination.

  1. 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.

  2. 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

  3. Determination of delta-aminolaevulinic acid in biological fluids by gas-liquid chromatography with electron-capture detection.

    PubMed Central

    Gorchein, A

    1984-01-01

    A derivative of delta-aminolaevulinic acid (AmLev), 2-methyl-3-acetyl-4-(3-propionic acid pentafluorobenzyl ester)pyrrole, with favourable properties for g.l.c. with electron-capture detection, was synthesized. Less than 1 pg could be detected on the column. 6-Amino-5-oxohexanoic acid formed the analogous derivative under similar conditions and was used as the internal standard in the development of a highly sensitive and specific assay for AmLev. The method has been applied to peripheral-venous and umbilical-cord plasma and to cerebrospinal fluid of normal and porphyric subjects. PMID:6547602

  4. 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

  5. Immunochemical Assays and Nucleic-Acid Detection Techniques for Clinical Diagnosis of Prostate Cancer

    PubMed Central

    Kanyong, Prosper; Rawlinson, Sean; Davis, James

    2016-01-01

    Prostate cancer (PCa) is a significant cause of morbidity and mortality and the most common cancer in men in Europe, North America, and some parts of Africa. The established methods for detecting PCa are normally based on tests using Prostate Specific Antigen (PSA) in blood, Prostate cancer antigen 3 (PCA3) in urine and tissue Alpha-methylacyl-CoA racemase (AMACR) as tumour markers in patient samples. Prior to the introduction of PSA in clinics, prostatic acid phosphatase (PAP) was the most widely used biomarker. An early diagnosis of PCa through the detection of these biomarkers requires the availability of simple, reliable, cost-effective and robust techniques. Immunoassays and nucleic acid detection techniques have experienced unprecedented growth in recent years and seem to be the most promising analytical tools. This growth has been driven in part by the surge in demand for near-patient-testing systems in clinical diagnosis. This article reviews immunochemical assays, and nucleic-acid detection techniques that have been used to clinically diagnose PCa. PMID:26958088

  6. 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...

  7. 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.

  8. 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.

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

    PubMed

    Cardador, María José; Gallego, Mercedes

    2014-05-01

    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.

  10. Compositions and methods for detecting Noonan syndrome

    DOEpatents

    Gelb, Bruce D.; Tartaglia, Marco; Pennacchio, Len

    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.

  11. [Simultaneous determination of 20 underivatized amino acids by high performance liquid chromatography-evaporative light-scattering detection].

    PubMed

    Wang, Yuhong; Shen, Keyu; Li, Peng; Zhou, Junyi; Chao, Yan

    2011-09-01

    An analytical method for the determination of underivatized amino acids was established with high performance liquid chromatography (HPLC) coupled with evaporative light-scattering detection (ELSD). A domestic evaporative light-scattering detector was used to determine underivatized amino acids. A BISCHOFF C18 AQ PLUS column and a solvent gradient elution with 0.2% heptafluorobutyric acid containing 0.1% trifluoroacetic acid as mobile phase A and methanol as mobile phase B were used. The temperature of the drift tube in ELSD was 40 degrees C and the flow rate of dry air was 2.5 L/min. The logarithm of the peak area and logarithm of the mass of each separated amino acid was in good linearity. The linear ranges of the 20 amino acids were from 30-300 mg/L. The limits of detection (S/N > 3) for the underivatized amino acids were from 24 ng to 100 ng. The average recoveries of the 20 amino acids were between 90.6% and 106.0%. This method and system are simple, rapid and accurate for the determination of underivatized amino acids. It can be used for the determination of underivatized amino acids in pharmaceutical, food and chemical industry fields.

  12. Determination of hippuric acid in human urine by ion chromatography with conductivity detection.

    PubMed

    Zhao, Fuyong; Wang, Zonghua; Wang, Hui; Ding, Mingyu

    2011-02-01

    A simple, rapid, precise and eco-friendly ion chromatography (IC) method for the determination of hippuric acid (HA) in human urine was proposed in this paper. The separation was carried out an anion exchange column with 2.0 mmol L⁻¹ Na₂CO₃ + 2.0 mmol L⁻¹ NaHCO₃ as mobile phase at the flow-rate 0.7 mL min⁻¹. A suppressed conductivity detector was used and the detection limit was 1.0 μg L⁻¹ (S/N=3) for hippuric acid. The analysis time for one run was 30 min under the optimized IC condition. The recovery of hippuric acid was 93.2-98.0% while the relative standard deviation (RSD) was 1.4-2.3% by seven measurements.

  13. [Bacteriocins: criteria, classification, characteristics, methods of detection].

    PubMed

    Blinkova, L P

    2003-01-01

    The review on bacteriocins of Gram negative and Gram positive bacteria. Criteria making it possible to regard antagonistic substances as bateriocins or bacteriocin-like substances and on their classification are presented. Examples of bacteriocins naming depending on the taxonomic position of the producer culture are given. Information on the physico-chemical and biological properties of bacteriocins and their purification is presented as well as on detection tools of bacteriocins in microorganisms and evaluation of the producer activity of the bacteriological culture.

  14. 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.

  15. 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.

  16. [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 PMID:26999859

  17. 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.

  18. High throughput automated colorimetric method for the screening of l-lactic acid producing microorganisms.

    PubMed

    Liaud, Nadège; Navarro, David; Vidal, Nicolas; Sigoillot, Jean-Claude; Raouche, Sana

    2014-01-01

    Lactic acid is a valuable and fully degradable organic acid with promising applications in poly-lactic acid production (Taskila S and Ojamo, 2013 [1]). Despite their efficiency, the cost of the current lactic acid bio-processes is still an obstacle to this application (Miller et al., 2011 [2]). To ameliorate lactic acid producing strains, researchers are using mutations and metabolic engineering techniques, as well as medium optimization. All these studies necessitate a good and high throughput screening method. Currently, researchers mostly use HPLC methods which often necessitate sample preparation, are not stereospecific and do not allow high throughput. To help optimizing l-lactic acid production, we developed a high throughput colorimetric method inspired by the blood l-lactic acid detection method used for diagnosis (Lin et al., 1999 [3]).•Two sequential enzymatic reactions using l-lactate oxidase, peroxidase and ABTS (2,2'-azino-di-[3-ethylbenzthiazoine-sulfonate]), a chromogenic peroxidase substrate, are used to quantify l-lactate between 13.8 and 90 mg/l.•The accuracy of the method was ascertained before automation.•The method was successfully applied for the direct determination of l-lactate content in fungal culture supernatants.

  19. Label-free visual detection of nucleic acids in biological samples with single-base mismatch detection capability.

    PubMed

    Song, Yanling; Zhang, Weiting; An, Yuan; Cui, Liang; Yu, Chundong; Zhu, Zhi; Yang, Chaoyong James

    2012-01-14

    We have combined an allosteric molecular beacon for target recognition and guanine-rich DNAzyme for signal amplification to develop a new platform for visual detection of nucleic acids with single-base mismatch detection capability. The fully DNA-structured platform can undergo color change in response to target DNA/RNA, which enables sensitive and selective visual detection in biological samples.

  20. A new method of pulse edge detection in low SNR

    NASA Astrophysics Data System (ADS)

    Fan, Xiaolei; Li, Tao; Su, Shaoying; Chen, Zengping

    2015-12-01

    In this paper, we propose a new pulse edge detection method based on short time Fourier transform (STFT) and difference of boxes (DOB) filter. Firstly, detect the coarse starting and ending positions in frequency domain after STFT. Then obtain the precise pulse edge through DOB filter. It achieves a better performance than the classical energy detection (ED) method, especially when signal to noise ratio (SNR) is low. Simulation results and real data application validate the effectiveness of the proposed method.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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

  7. 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.

  8. Abnormality degree detection method using negative potential field group detectors

    NASA Astrophysics Data System (ADS)

    Zhang, Hongli; Liu, Shulin; Li, Dong; Shi, Kunju; Wang, Bo; Cui, Jiqiang

    2015-09-01

    Online monitoring methods have been widely used in many major devices, however the normal and abnormal states of equipment are estimated mainly based on the monitoring results whether monitored parameters exceed the setting thresholds. Using these monitoring methods may cause serious false positive or false negative results. In order to precisely monitor the state of equipment, the problem of abnormality degree detection without fault sample is studied with a new detection method called negative potential field group detectors(NPFG-detectors). This method achieves the quantitative expression of abnormality degree and provides the better detection results compared with other methods. In the process of Iris data set simulation, the new algorithm obtains the successful results in abnormal detection. The detection rates for 3 types of Iris data set respectively reach 100%, 91.6%, and 95.24% with 50% training samples. The problem of Bearing abnormality degree detection via an abnormality degree curve is successfully solved.

  9. 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.

  10. 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. PMID:27265004

  11. 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.

  12. Rapid determination of nonaromatic organic acids in honey by capillary zone electrophoresis with direct ultraviolet detection.

    PubMed

    Mato, Inés; Huidobro, José F; Simal-Lozano, Jesús; Sancho, M Teresa

    2006-03-01

    A rapid capillary zone electrophoresis (CZE) method with direct ultraviolet (UV) detection has been set up and developed to determine the most important nonaromatic organic acids in honey with a really simple treatment of the sample. The determination of oxalic, formic, malic, succinic, pyruvic, acetic, lactic, citric, and gluconic acids has been carried out in 4 min. The electrolyte composition was phosphate as the carrier buffer (7.5 mM NaH(2)PO(4) and 2.5 mM Na(2)HPO(4)), 2.5 mM tetradecyltrimethylammonium hydroxide (TTAOH) as electroosmotic flow modifier, and 0.24 mM CaCl(2) as selectivity modifier, with the pH adjusted at 6.40 constant value. The running voltage was -25 kV at a thermostated temperature of 25 degrees C. The injections were performed in hydrodynamic mode (30 s), and the detection mode was UV direct at 185 nm. Validation parameters of the method as detection and quantification limits, linearity, precision (repeatability and reproducibility), and recovery were also studied. The advantages related to the technique such as simplicity, short analysis times, and low consumption of chemicals as well as the good validation parameters obtained for this method permit it to be considered as adequate for routine analysis in honey.

  13. 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

  14. 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.

  15. Multimode nondestructive detecting method for high-speed rail defects

    NASA Astrophysics Data System (ADS)

    Sun, Mingjian; Cheng, Xingzhen; Wan, Guangnan; Liu, Ting; Fu, Ying; Wang, Yan

    2015-11-01

    It is very important to detect the surface defects of the high-speed rail for security concerns. A multimode detecting method, which integrates high resolution of optical image, high precision of photoacoustic detection and strong penetration of ultrasound detecting, is proposed for the rail defect detection. Utilizing the surface defect characteristics obtained from optical signal, the photoacoustic and ultrasound scanning region could be determined, and rail shallow and internal defect characteristics can be acquired subsequently. Eventually, fusing three modal signals mentioned above, the information of the entire rail defect, including type, extension trend and depth can be detected. It has been proved that the multimode method can improve the detecting efficiency, and enlarge the detection range in the meantime.

  16. [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

  17. Mass spectrometric detection of the amino acid sequence polymorphism of the hepatitis C virus antigen.

    PubMed

    Kaysheva, A L; Ivanov, Yu D; Frantsuzov, P A; Krohin, N V; Pavlova, T I; Uchaikin, V F; Konev, V А; Kovalev, O B; Ziborov, V S; Archakov, A I

    2016-03-01

    A method for detection and identification of the hepatitis C virus antigen (HCVcoreAg) in human serum with consideration for possible amino acid substitutions is proposed. The method is based on a combination of biospecific capturing and concentrating of the target protein on the surface of the chip for atomic force microscope (AFM chip) with subsequent protein identification by tandem mass spectrometric (MS/MS) analysis. Biospecific AFM-capturing of viral particles containing HCVcoreAg from serum samples was performed by use of AFM chips with monoclonal antibodies (anti-HCVcore) covalently immobilized on the surface. Biospecific complexes were registered and counted by AFM. Further MS/MS analysis allowed to reliably identify the HCVcoreAg in the complexes formed on the AFM chip surface. Analysis of MS/MS spectra, with the account taken of the possible polymorphisms in the amino acid sequence of the HCVcoreAg, enabled us to increase the number of identified peptides.

  18. Formation, reactivity and detection of protein sulfenic acids

    PubMed Central

    Kettenhofen, Nicholas J.; Wood, Matthew J.

    2010-01-01

    It has become clear in recent decades that the post-translational modification of protein cysteine residues is a crucial regulatory event in biology. Evidence supports the reversible oxidation of cysteine thiol groups as a mechanism of redox-based signal transduction while the accumulation of proteins with irreversible thiol oxidations is a hallmark of stress-induced cellular damage. The initial formation of cysteine sulfenic acid (SOH) derivatives, along with the reactive properties of this functional group, serves as a crossroads whereby the local redox environment may dictate the progression of either regulatory or pathological outcomes. Protein-SOH are established as transient intermediates in the formation of more stable cysteine oxidation products both under basal conditions and in response to several redox-active extrinsic compounds. This review details both direct and multi-step chemical routes proposed to generate protein-SOH, the spectrum of secondary reactions that may follow their initial formation and the arsenal of experimental tools available for their detection. Both the pioneering studies that have provided a framework for our current understanding of protein-SOH as well as state-of-the-art proteomic strategies designed for global assessments of this post-translational modification are highlighted. PMID:20845928

  19. 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.

  20. Colorimetric Detection of Cadmium Ions Using DL-Mercaptosuccinic Acid-Modified Gold Nanoparticles.

    PubMed

    Chen, Na; Chen, Jun; Yang, Jing-Hua; Bai, Lian-Yang; Zhang, Yu-Ping

    2016-01-01

    A colorimetric assay has been developed for detection of Cd²⁺ utilizing DL-mercaptosuccinic acid-modified gold nanoparticles (MSA-AuNPs). The method showed good selectivity for Cd²⁺ over other metal ions. As a result, the linear relationships (r > 0.9606) between concentration 0.07 mM and 0.20 mM for cadmium ion were obtained. The detection limit was as low as 0.07 mM by the naked eye. The effect of pH on the aggregation was optimized. The MSA-AuNPs probe could be used to detect Cd²⁺ in an aqueous solution based on the aggregation-induced color change of MSA-AuNPs. PMID:27398533

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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

  7. 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.

  8. Laser pulse detection method and apparatus

    NASA Astrophysics Data System (ADS)

    Goss, W.; Janesick, J. R.

    1984-02-01

    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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. A Sensitive Peptide Nucleic Acid Probe Assay for Detection of BRAF V600 Mutations in Melanoma.

    PubMed

    Chen, Tai-Long; Chang, John Wen-Cheng; Hsieh, Jia-Juan; Cheng, Hsin-Yi; Chiou, Chiuan-Chian

    Mutated v-Raf murine sarcoma viral oncogene homolog B (BRAF) is an important biomarker for the prediction of therapeutic efficacy of several anticancer drugs. The detection of BRAF mutation faces two challenges: Firstly, there are multiple types of mutations, and secondly, tumor samples usually contain various amounts of wild-type, normal tissues. Here, we describe a newly established method for sensitive detection of multiple types of BRAF V600 mutations in excess wild-type background. The method introduced a fluorophore-tagged peptide nucleic acid (PNA) to serve as both polymerase chain reaction (PCR) clamp and sensor probe, which inhibited the amplification of wild-type templates during PCR and revealed multiple types of mutant signals during melting analysis. We demonstrated the design and optimization process of the method, and applied it in the detection of BRAF mutations in 49 melanoma samples. This PNA probe assay method detected three types of mutations in 17 samples, and was much more sensitive than conventional PCR plus Sanger sequencing. PMID:27566656

  14. 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...

  15. 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.

  16. 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.

  17. Molecular detection of harmful algal blooms (HABs) using locked nucleic acids and bead array technology

    PubMed Central

    Diaz, Mara R.; Jacobson, James W.; Goodwin, Kelly D.; Dunbar, Sherry A.; Fell, Jack W.

    2010-01-01

    Harmful algal blooms (HABs) are a serious public health risk in coastal waters. As the intensity and frequency of HABs continue to rise, new methods of detection are needed for reliable identification. Herein, we developed a high-throughput, multiplex, bead array technique for the detection of the dinoflagellates Karenia brevis and Karenia mikimotoi. The method combined the Luminex detection system with two novel technologies: locked nucleic acid–modified oligonucleotides (LNA) and Mirus Label IT® nucleic acid technology. To study the feasibility of the method, we evaluated the performance of modified and unmodified LNA probes with amplicon targets that were biotin labeled with two different strategies: direct chemical labeling (Mirus Label IT) versus enzymatic end-labeling (single biotinylated primer). The results illustrated that LNA probes hybridized to complementary single-stranded DNA with better affinity and displayed higher fluorescence intensities than unmodified oligonucleotide DNA probes. The latter effect was more pronounced when the assay was carried out at temperatures above 53°C degree. As opposed to the enzymatic 5′ terminal labeling technique, the chemical-labeling method enhanced the level of fluorescence by as much as ~83%. The detection limits of the assay, which were established with LNA probes and Mirus Label IT system, ranged from 0.05 to 46 copies of rRNA. This high-throughput method, which represents the first molecular detection strategy to integrate Luminex technology with LNA probes and Mirus Label IT, can be adapted for the detection of other HABs and is well suited for the monitoring of red tides at pre-blooming and blooming conditions. PMID:21165155

  18. 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.

  19. 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.

  20. Analytical Methods for Secondary Metabolite Detection.

    PubMed

    Taibon, Judith; Strasser, Hermann

    2016-01-01

    The entomopathogenic fungi Metarhizium brunneum, Beauveria bassiana, and B. brongniartii are widely applied as biological pest control agent in OECD countries. Consequently, their use has to be flanked by a risk management approach, which includes the need to monitor the fate of their relevant toxic metabolites. There are still data gaps claimed by regulatory authorities pending on their identification and quantification of relevant toxins or secondary metabolites. In this chapter, analytical methods are presented allowing the qualitative and quantitative analysis of the relevant toxic B. brongniartii metabolite oosporein and the three M. brunneum relevant destruxin (dtx) derivatives dtx A, dtx B, and dtx E. PMID:27565501

  1. Analytical Methods for Secondary Metabolite Detection.

    PubMed

    Taibon, Judith; Strasser, Hermann

    2016-01-01

    The entomopathogenic fungi Metarhizium brunneum, Beauveria bassiana, and B. brongniartii are widely applied as biological pest control agent in OECD countries. Consequently, their use has to be flanked by a risk management approach, which includes the need to monitor the fate of their relevant toxic metabolites. There are still data gaps claimed by regulatory authorities pending on their identification and quantification of relevant toxins or secondary metabolites. In this chapter, analytical methods are presented allowing the qualitative and quantitative analysis of the relevant toxic B. brongniartii metabolite oosporein and the three M. brunneum relevant destruxin (dtx) derivatives dtx A, dtx B, and dtx E.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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)

  8. Method for distinctive estimation of stored acidity forms in acid mine wastes.

    PubMed

    Li, Jun; Kawashima, Nobuyuki; Fan, Rong; Schumann, Russell C; Gerson, Andrea R; Smart, Roger St C

    2014-10-01

    Jarosites and schwertmannite can be formed in the unsaturated oxidation zone of sulfide-containing mine waste rock and tailings together with ferrihydrite and goethite. They are also widely found in process wastes from electrometallurgical smelting and metal bioleaching and within drained coastal lowland soils (acid-sulfate soils). These secondary minerals can temporarily store acidity and metals or remove and immobilize contaminants through adsorption, coprecipitation, or structural incorporation, but release both acidity and toxic metals at pH above about 4. Therefore, they have significant relevance to environmental mineralogy through their role in controlling pollutant concentrations and dynamics in contaminated aqueous environments. Most importantly, they have widely different acid release rates at different pHs and strongly affect drainage water acidity dynamics. A procedure for estimation of the amounts of these different forms of nonsulfide stored acidity in mining wastes is required in order to predict acid release rates at any pH. A four-step extraction procedure to quantify jarosite and schwertmannite separately with various soluble sulfate salts has been developed and validated. Corrections to acid potentials and estimation of acid release rates can be reliably based on this method.

  9. 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. PMID:25399119

  10. 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.

  11. Using statistical methods and genotyping to detect tuberculosis outbreaks

    PubMed Central

    2013-01-01

    Background Early identification of outbreaks remains a key component in continuing to reduce the burden of infectious disease in the United States. Previous studies have applied statistical methods to detect unexpected cases of disease in space or time. The objectives of our study were to assess the ability and timeliness of three spatio-temporal methods to detect known outbreaks of tuberculosis. Methods We used routinely available molecular and surveillance data to retrospectively assess the effectiveness of three statistical methods in detecting tuberculosis outbreaks: county-based log-likelihood ratio, cumulative sums, and a spatial scan statistic. Results Our methods identified 8 of the 9 outbreaks, and 6 outbreaks would have been identified 1–52 months (median = 10 months) before local public health authorities identified them. Assuming no delays in data availability, 46 (59.7%) of the 77 patients in the 9 outbreaks were identified after our statistical methods would have detected the outbreak but before local public health authorities became aware of the problem. Conclusions Statistical methods, when applied retrospectively to routinely collected tuberculosis data, can successfully detect known outbreaks, potentially months before local public health authorities become aware of the problem. The three methods showed similar results; no single method was clearly superior to the other two. Further study to elucidate the performance of these methods in detecting tuberculosis outbreaks will be done in a prospective analysis. PMID:23497235

  12. Separation and determination of amino acids by micellar electrokinetic chromatography coupling with novel multiphoton excited fluorescence detection.

    PubMed

    Chen, Sheng; Xu, Youzhi; Xu, Fei; Feng, Xiaojun; Du, Wei; Luo, Qingming; Liu, Bi-Feng

    2007-08-31

    In this article, it was demonstrated that separation and determination of 20 amino acids were accomplished by micellar electrokinetic chromatography (MEKC) coupling with novel multiphoton excited fluorescence (MPEF) detection method. Different from MPEF achieved by expensive fs laser, continuous wave (CW) diode laser of ultra-low cost was uniquely employed in our MPEF system. Amino acids were fluorescently labeled with fluorescein isothiocyanate (FITC), and were subjected to sodium dodecyl sulfate (SDS)-based MEKC separation and CW-based MPEF detection. The result was compared with that by single photon excited fluorescence (SPEF), which indicated that MPEF had the advantages of better mass detectability and higher separation selectivity over SPEF. Quantitative analysis was performed and revealed linear dynamic range of over 2 orders of magnitude, with mass detection limit down to ymole level. To evaluate the reliability, this method was successfully applied for analyzing a commercial nutrition supplement liquid.

  13. 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.

  14. 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. PMID:24591747

  15. Food allergen detection methods: a coordinated approach.

    PubMed

    Goodwin, Philip R

    2004-01-01

    The levels (1-2%) and increasing severity of allergic responses to food in the adult population are well documented, as is the phenomenon of even higher (3-8%) and apparently increasing incidence in children, albeit that susceptibility decreases with age. Problematic foods include peanut, milk, eggs, tree nuts, and sesame, but the list is growing as awareness continues to rise. The amounts of such foods that can cause allergic reactions is difficult to gauge; however, the general consensus is that ingestion of low parts per million is sufficient to cause severe reactions in badly affected individuals. Symptoms can rapidly-within minutes-progress from minor discomfort to severe, even life-threatening anaphylactic shock in those worst affected. Given the combination of high incidence of atopy, potential severity of response, and apparently widespread instances of "hidden" allergens in the food supply, it is not surprising that this issue is increasingly subject to legislative and regulatory scrutiny. In order to assist in the control of allergen levels in foods to acceptable levels, analysts require a combination of test methods, each designed to produce accurate, timely, and cost-effective analytical information. Such information contributes significantly to Hazard Analysis Critical Control Point programs to determine food manufacturers' risk and improves the accuracy of monitoring and surveillance by food industry, commercial, and enforcement laboratories. Analysis thereby facilitates improvements in compliance with labeling laws with concomitant reductions in risks to atopic consumers. This article describes a combination of analytical approaches to fulfill the various needs of these 3 analytical communities.

  16. 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.

  17. 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. PMID:25571382

  18. 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.

  19. 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...

  20. A novel method for determining peroxisomal fatty acid β-oxidation.

    PubMed

    Morita, Masashi; Matsumoto, Shun; Okazaki, Airi; Tomita, Kaito; Watanabe, Shiro; Kawaguchi, Kosuke; Minato, Daishiro; Matsuya, Yuji; Shimozawa, Nobuyuki; Imanaka, Tsuneo

    2016-09-01

    The purpose of this study is to establish an assay method to screen for chemical compounds that stimulate peroxisomal fatty acid β-oxidation activity in X-linked adrenoleukodystropy (X-ALD) fibroblasts. In this investigation, we used 12-(1-pyrene)dodecanoic acid (pyrene-C12:0), a fluorescent fatty acid analog, as a substrate for fatty acid β-oxidation. When human skin fibroblasts were incubated with pyrene-C12:0, β-oxidation products such as pyrene-C10:0 and pyrene-C8:0 were generated time-dependently. These β-oxidation products were scarcely detected in the fibroblasts from patients with Zellweger syndrome, a peroxisomal biogenesis disorder. In contrast, in fibroblasts with mitochondrial carnitine-acylcarnitine translocase deficiency, the β-oxidation products were detected at a level similar to control fibroblasts. These results indicate that the β-oxidation of pyrene-C12:0 takes place in peroxisomes, but not mitochondria, so pyrene-C12:0 is useful for measuring peroxisomal fatty acid β-oxidation activity. In X-ALD fibroblasts, the β-oxidation activity for pyrene-C12:0 was approximately 40 % of control fibroblasts, which is consistent with previous results using [1-(14)C]lignoceric acid as the substrate. The present study provides a convenient procedure for screening chemical compounds that stimulate the peroxisomal fatty acid β-oxidation in X-ALD fibroblasts.

  1. Wheat gluten amino acid composition analysis by high-performance anion-exchange chromatography with integrated pulsed amperometric detection.

    PubMed

    Rombouts, Ine; Lamberts, Lieve; Celus, Inge; Lagrain, Bert; Brijs, Kristof; Delcour, Jan A

    2009-07-17

    A simple accurate method for determining amino acid composition of wheat gluten proteins and their gliadin and glutenin fractions using high-performance anion-exchange chromatography with integrated pulsed amperometric detection is described. In contrast to most conventional methods, the analysis requires neither pre- or post-column derivatization, nor oxidation of the sample. It consists of hydrolysis (6.0M hydrochloric acid solution at 110 degrees C for 24h), evaporation of hydrolyzates (110 degrees C), and chromatographic separation of the liberated amino acids. Correction factors (f) accounted for incomplete cleavage of peptide bonds involving Val (f=1.07) and Ile (f=1.13) after hydrolysis for 24h and for Ser (f=1.32) losses during evaporation. Gradient conditions including an extra eluent (0.1M acetic acid solution) allowed multiple sequential sample analyses without risk of Glu contamination on the anion-exchange column. While gluten amino acid compositions by the present method were mostly comparable to those obtained by a conventional method involving oxidation, acid hydrolysis and post-column ninhydrin derivatization, the latter method underestimated Tyr, Val and Ile levels. Results for the other amino acids obtained by the different methods were linearly correlated (r>0.99, slope=1.03).

  2. 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.

  3. Molecular detection of human adenovirus in sediment using a direct detection method compared to the classical polyethylene glycol precipitation.

    PubMed

    Staggemeier, Rodrigo; Bortoluzzi, Marina; da Silva Heck, Tatiana Moraes; da Silva, Tiago; Spilki, Fernando Rosado; de Matos Almeida, Sabrina Esteves

    2015-03-01

    Various effective methods have been developed to measure the concentration of viruses in sediment samples. However, there is need to standardize less laborious and simpler techniques. The objective of the present study was to compare two different methods to measure the concentration of viruses in soil samples. The use of polyethylene glycol (PEG) was compared with a direct extraction of viral nucleic acids from the samples diluted in modified Eagle's minimal essential medium (E-MEM). The presence of adenovirus in the samples was detected by real-time quantitative polymerase chain reaction (qPCR). Only six samples (30%) were positive for adenovirus when PEG technique was used. The direct method showed 16 (80%) samples positive for adenovirus. Therefore, direct detection (i.e. without previous concentration) demonstrated a higher rate of detection, better effectiveness, and shorter execution time. Furthermore, direct detection uses reagents that are often readily available in virology laboratories. Thus, it is an attractive alternative to other methods of detection of virus particles in sediments.

  4. Detection of Vibrio cholerae by real-time nucleic acid sequence-based amplification.

    PubMed

    Fykse, Else M; Skogan, Gunnar; Davies, William; Olsen, Jaran Strand; Blatny, Janet M

    2007-03-01

    A multitarget molecular beacon-based real-time nucleic acid sequence-based amplification (NASBA) assay for the specific detection of Vibrio cholerae has been developed. The genes encoding the cholera toxin (ctxA), the toxin-coregulated pilus (tcpA; colonization factor), the ctxA toxin regulator (toxR), hemolysin (hlyA), and the 60-kDa chaperonin product (groEL) were selected as target sequences for detection. The beacons for the five different genetic targets were evaluated by serial dilution of RNA from V. cholerae cells. RNase treatment of the nucleic acids eliminated all NASBA, whereas DNase treatment had no effect, showing that RNA and not DNA was amplified. The specificity of the assay was investigated by testing several isolates of V. cholerae, other Vibrio species, and Bacillus cereus, Salmonella enterica, and Escherichia coli strains. The toxR, groEL, and hlyA beacons identified all V. cholerae isolates, whereas the ctxA and tcpA beacons identified the O1 toxigenic clinical isolates. The NASBA assay detected V. cholerae at 50 CFU/ml by using the general marker groEL and tcpA that specifically indicates toxigenic strains. A correlation between cell viability and NASBA was demonstrated for the ctxA, toxR, and hlyA targets. RNA isolated from different environmental water samples spiked with V. cholerae was specifically detected by NASBA. These results indicate that NASBA can be used in the rapid detection of V. cholerae from various environmental water samples. This method has a strong potential for detecting toxigenic strains by using the tcpA and ctxA markers. The entire assay including RNA extraction and NASBA was completed within 3 h.

  5. A novel method for determining target detection thresholds

    NASA Astrophysics Data System (ADS)

    Grossman, S.

    2015-05-01

    Target detection is the act of isolating objects of interest from the surrounding clutter, generally using some form of test to include objects in the found class. However, the method of determining the threshold is overlooked relying on manual determination either through empirical observation or guesswork. The question remains: how does an analyst identify the detection threshold that will produce the optimum results? This work proposes the concept of a target detection sweet spot where the missed detection probability curve crosses the false detection curve; this represents the point at which missed detects are traded for false detects in order to effect positive or negative changes in the detection probability. ROC curves are used to characterize detection probabilities and false alarm rates based on empirically derived data. It identifies the relationship between the empirically derived results and the first moment statistic of the histogram of the pixel target value data and then proposes a new method of applying the histogram results in an automated fashion to predict the target detection sweet spot at which to begin automated target detection.

  6. Development of a rapid and simple method for detection of protein contaminants in carmine.

    PubMed

    Nakayama, Norihisa; Ohtsu, Yutaka; Maezawa-Kase, Daisuke; Sano, Ken-Ichi

    2015-01-01

    Protein contaminants in carmine can cause dyspnea and anaphylactic reactions in users and consumers of products containing this pigment. The method generally used for detection of proteins in carmine has low reproducibility and is time-consuming. In this study, a rapid, simple, and highly reproducible method was developed for the detection of protein contaminants in carmine. This method incorporates acidic protein denaturation conditions and ultrafiltration. To prevent protein aggregation, sodium dodecyl sulfate containing gel electrophoresis running buffer was used for dispersing the carmine before filtration. An ultrafiltration device was used to separate the protein contaminants from carminic acid in the carmine solution. Two ultrafiltration devices were compared, and a cylindrical device containing a modified polyethersulfone membrane gave the best results. The method had high reproducibility. PMID:25892994

  7. Development of a Rapid and Simple Method for Detection of Protein Contaminants in Carmine

    PubMed Central

    Nakayama, Norihisa; Ohtsu, Yutaka; Maezawa-Kase, Daisuke; Sano, Ken-Ichi

    2015-01-01

    Protein contaminants in carmine can cause dyspnea and anaphylactic reactions in users and consumers of products containing this pigment. The method generally used for detection of proteins in carmine has low reproducibility and is time-consuming. In this study, a rapid, simple, and highly reproducible method was developed for the detection of protein contaminants in carmine. This method incorporates acidic protein denaturation conditions and ultrafiltration. To prevent protein aggregation, sodium dodecyl sulfate containing gel electrophoresis running buffer was used for dispersing the carmine before filtration. An ultrafiltration device was used to separate the protein contaminants from carminic acid in the carmine solution. Two ultrafiltration devices were compared, and a cylindrical device containing a modified polyethersulfone membrane gave the best results. The method had high reproducibility. PMID:25892994

  8. Determination of linoleic acid in toothpaste by gas chromatography with flame ionization detection.

    PubMed

    Wejnerowska, Grazyna; Gackowska, Alicja; Gaca, Jerzy

    2008-06-01

    A new method for the determination of linoleic acid (LA) in toothpaste by a routine analysis has been proposed. Studies were based on the ISO 5509 procedure, which was modified for the purpose of LA determination in the toothpaste. Gas chromatography (GC) was employed for the qualitative and quantitative determination of linoleic acid methyl ester. The content of LA (5.31%) in sunflower oil added to the toothpaste composition (0.5%) was determined, and then the optimization studies for the determination of LA in the toothpaste samples were carried out. The relative standard deviation (RSD) of the procedure developed was 9.96% (n = 9). The quantitative analysis showed that the content of LA in the toothpaste samples studied was 0.0258 +/- 0.0011%. The detection limit of LA in toothpaste was approximately 0.001%.

  9. 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).

  10. 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

    2015-01-01

    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). PMID:26198220

  11. 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.

  12. Method to attenuate U(VI) mobility in acidic waste plumes using humic acids.

    PubMed

    Wan, Jiamin; Dong, Wenming; Tokunaga, Tetsu K

    2011-03-15

    Acidic uranium (U) groundwater 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/year) show that desorption of U and HA were nondetectable 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 indicate 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.

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

    PubMed

    Wan, Jiamin; Dong, Wenming; Tokunaga, Tetsu K

    2011-03-15

    Acidic uranium (U) groundwater 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/year) show that desorption of U and HA were nondetectable 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 indicate 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. PMID:21319737

  14. Two methods for the separation of monounsaturated octadecenoic acid isomers.

    PubMed

    Villegas, C; Zhao, Y; Curtis, J M

    2010-01-29

    The identification and quantification of complex mixtures of cis and trans octadecenoic (18:1) fatty acid isomers presents a major challenge for conventional one-dimensional GC/FID analysis of their methyl esters. We have compared the use of two methods to achieve optimized separations of positional and geometrical octadecenoic fatty acid isomers-comprehensive two-dimensional gas chromatography (GCxGC), and silver ion high performance liquid chromatography interfaced to atmospheric pressure photoionization (APPI) mass spectrometry. Nine isomers of octadecenoic acid methyl ester were well separated on a single silver ion column with a mobile phase of 0.018% acetonitrile and 0.18% isopropanol in hexane. Reproducible retention times were obtained with relative standard deviations of around 1% over 5 injections. The extra selectivity and reproducibility afforded by APPI-MS, together with the wide separation of cis and trans isomers by silver ion chromatography, resulted in a promising method for measurement of octadecenoic acid FAME. The GCxGC separation was performed using various column combinations, and optimal separation was obtained by coupling an ionic liquid column (Supelco SLB-IL100 [1,9-di(3-vinyl-imidazolium) nonane bis(trifluoromethyl) sulfonyl imidate]) in the first dimension with a SGE BPX50 (50% phenyl polysilphenylene-siloxane) in the second dimension. These methods have been applied to the analysis of octadecenoic acid in milk and beef fat. PMID:20022011

  15. 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.

  16. 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.

  17. 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. PMID:27325503

  18. 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.

  19. 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.

  20. Wheat gluten amino acid analysis by high-performance anion-exchange chromatography with integrated pulsed amperometric detection.

    PubMed

    Rombouts, Ine; Lagrain, Bert; Lamberts, Lieve; Celus, Inge; Brijs, Kristof; Delcour, Jan A

    2012-01-01

    This chapter describes an accurate and user-friendly method for determining amino acid composition of wheat gluten proteins and their gliadin and glutenin fractions. The method consists of hydrolysis of the peptide bonds in 6.0 M hydrochloric acid solution at 110°C for 24 h, followed by evaporation of the acid and separation of the free amino acids by high-performance anion-exchange chromatography with integrated pulsed amperometric detection. In contrast to conventional methods, the analysis requires neither pre- or postcolumn derivatization, nor a time-consuming oxidation or derivatization step prior to hydrolysis. Correction factors account for incomplete release of Val and Ile even after hydrolysis for 24 h, and for losses of Ser during evaporation. Gradient conditions including an extra eluent allow multiple sequential sample analyses without risk of Glu accumulation on the anion-exchange column which otherwise would result from high Gln levels in gluten proteins.

  1. Multicommutated stepwise injection determination of ascorbic acid in medicinal plants and food samples by capillary zone electrophoresis ultraviolet detection.

    PubMed

    Falkova, Marina T; Bulatov, Andrey V; Pushina, Maria O; Ekimov, Aleksey A; Alekseeva, Galina M; Moskvin, Leonid N

    2015-02-01

    An automation of the extraction of analytes from solid samples into the aqueous phase based on multicommutated stepwise injection analysis concept has been suggested. The feasibility of the approach has been demonstrated by determination of ascorbic acid as model analyte. The method includes automated extraction of ascorbic acid from solid sample into borate buffer solution pH 8 in mixing chamber during vigorous mixing by nitrogen stream, and subsequent detection by capillary zone electrophoresis at 254 nm. The method has a linear range of 0.1-5.0 mg g(-1) for ascorbic acid with the LOD of 0.03 mg g(-1). The sample throughput was 7 h(-1). This method was applied for determination of ascorbic acid in various medicinal plants and food samples. PMID:25435231

  2. 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.

  3. 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.

  4. Comparison study of seizure detection using stationary and nonstationary methods.

    PubMed

    Li, Ying; Hsin, Yue-Loong; Liu, Wentai

    2014-01-01

    We present an accurate seizure detection algorithm, and make a detailed comparison of two frequency analysis methods: a widely used stationary method - Fast Fourier Transform (FFT) and a relatively new nonstationary method - Hilbert-Huang Transform (HHT). Two public databases and one our own database were tested. The results show that our algorithm has very high accuracy compared with the state-of-the-art. More interestingly, it shows that the nonstationary method HHT offers better performance than the stationary method FFT in seizure detection. Therefore we propose that we should pay attention to the nonstationarity of EEG signal, since the "stationary assumption" may introduce some inaccuracy.

  5. Bioanalytical Application of Amino Acid Detection by Capillary Electrophoresis.

    PubMed

    Fico, Daniela; Pennetta, Antonio; De Benedetto, Giuseppe E

    2016-01-01

    This chapter illustrates the usefulness of capillary electrophoresis (CE) for the analysis of amino acids, and both normal and chiral separations are covered. In order to provide a general description of the main results and challenges in the biomedical field, some relevant applications and reviews on CE of amino acids are tabulated. Furthermore, some detailed experimental procedures are shown, regarding the CE analysis of amino acids in body fluids, in microdialysate, and released upon hydrolysis of proteins. In particular, the protocols will deal with the following compounds: (1) underivatized aminoacids in blood; (2) γ-Aminobutyric acid, glutamate, and L-Aspartate derivatized with Naphthalene-2,3-dicarboxaldehyde; (3) hydrolysate from bovine serum albumine derivatized with phenylisothiocyanate. By examining these applications on real matrices, the capillary electrophoresis efficiency as tool for Amino Acid analysis can be ascertained. PMID:27645741

  6. Bayesian methods for outliers detection in GNSS time series

    NASA Astrophysics Data System (ADS)

    Qianqian, Zhang; Qingming, Gui

    2013-07-01

    This article is concerned with the problem of detecting outliers in GNSS time series based on Bayesian statistical theory. Firstly, a new model is proposed to simultaneously detect different types of outliers based on the conception of introducing different types of classification variables corresponding to the different types of outliers; the problem of outlier detection is converted into the computation of the corresponding posterior probabilities, and the algorithm for computing the posterior probabilities based on standard Gibbs sampler is designed. Secondly, we analyze the reasons of masking and swamping about detecting patches of additive outliers intensively; an unmasking Bayesian method for detecting additive outlier patches is proposed based on an adaptive Gibbs sampler. Thirdly, the correctness of the theories and methods proposed above is illustrated by simulated data and then by analyzing real GNSS observations, such as cycle slips detection in carrier phase data. Examples illustrate that the Bayesian methods for outliers detection in GNSS time series proposed by this paper are not only capable of detecting isolated outliers but also capable of detecting additive outlier patches. Furthermore, it can be successfully used to process cycle slips in phase data, which solves the problem of small cycle slips.

  7. [Standardization of technical methods for apple fluorescence canopy spectral detection].

    PubMed

    Zhu, Xi-Cun; Zhao, Geng-Xing; Lei, Tong; Wang, Ling; Dong, Fang; Wang, Jing-An

    2010-06-01

    Aiming at spectral detection of apple fluorescence canopy, the present paper carried out spectral detection tests under different weather conditions, different detection times, and different detection heights and angles to apple canopy in the two years of 2008 and 2009, so as to analyze impacts of these factors on apple canopy spectral characteristics and explore standardized spectral detection methods for apple fluorescence canopy. The results indicated the regularity in spectral reflectance of apple fluorescence canopy to a certain degree under different conditions, especially in the 760-1 350 nm near-infrared bands. The authors found that canopy spectral reflectance declined along with the decrease in sunshine and it is appropriate to detect canopy spectrum in sunny days with few clouds. In addition, spectral reflectance tended to be stable when the wind scale was below grade 2. The discrepancy of canopy spectra is small during the time period from 10:00 to 15:00 of a day compared to that of other times. For maintaining stable spectral curves, the height of detector to apple canopy needed to be adjusted to cover the whole canopy within the field of view according to detection angle of the detector. The vertical or approximately vertical detection was the best for canopy spectral reflectance acquisition. The standardization of technical methods of spectral detection for apple fluorescence canopy was proposed accordingly, which provided theoretical references for spectral detection and information extraction of apple tree canopy.

  8. 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.

  9. 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.

  10. Entropy Beacon: A Hairpin-Free DNA Amplification Strategy for Efficient Detection of Nucleic Acids.

    PubMed

    Lv, Yifan; Cui, Liang; Peng, Ruizi; Zhao, Zilong; Qiu, Liping; Chen, Huapei; Jin, Cheng; Zhang, Xiao-Bing; Tan, Weihong

    2015-12-01

    Here, we propose an efficient strategy for enzyme- and hairpin-free nucleic acid detection called an entropy beacon (abbreviated as Ebeacon). Different from previously reported DNA hybridization/displacement-based strategies, Ebeacon is driven forward by increases in the entropy of the system, instead of free energy released from new base-pair formation. Ebeacon shows high sensitivity, with a detection limit of 5 pM target DNA in buffer and 50 pM in cellular homogenate. Ebeacon also benefits from the hairpin-free amplification strategy and zero-background, excellent thermostability from 20 °C to 50 °C, as well as good resistance to complex environments. In particular, based on the huge difference between the breathing rate of a single base pair and two adjacent base pairs, Ebeacon also shows high selectivity toward base mutations, such as substitution, insertion, and deletion and, therefore, is an efficient nucleic acid detection method, comparable to most reported enzyme-free strategies.

  11. 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

  12. Quantitative detection of hepatitis B virus DNA by real-time nucleic acid sequence-based amplification with molecular beacon detection.

    PubMed

    Yates, S; Penning, M; Goudsmit, J; Frantzen, I; van de Weijer, B; van Strijp, D; van Gemen, B

    2001-10-01

    We have developed a hepatitis B virus (HBV) DNA detection and quantification system based on amplification with nucleic acid sequence-based amplification (NASBA) technology and real-time detection with molecular beacon technology. NASBA is normally applied to amplify single-stranded target RNA, producing RNA amplicons. In this work we show that with modifications like primer design, sample extraction method, and template denaturation, the NASBA technique can be made suitable for DNA target amplification resulting in RNA amplicons. A major advantage of our assay is the one-tube, isothermal nature of the method, which allows high-throughput applications for nucleic acid detection. The homogeneous real-time detection allows a closed-tube format of the assay, avoiding any postamplification handling of amplified material and therefore minimizing the risk of contamination of subsequent reactions. The assay has a detection range of 10(3) to 10(9) HBV DNA copies/ml of plasma or serum (6 logs), with good reproducibility and precision. Compared with other HBV DNA assays, our assay provides good sensitivity, a wide dynamic range, and high-throughput applicability, making it a viable alternative to those based on other amplification or detection methods.

  13. 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.

  14. 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.

  15. Passive background correction method for spatially resolved detection

    DOEpatents

    Schmitt, Randal L.; Hargis, Jr., Philip J.

    2011-05-10

    A method for passive background correction during spatially or angularly resolved detection of emission that is based on the simultaneous acquisition of both the passive background spectrum and the spectrum of the target of interest.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. Study of TATP: method for determination of residual acids in TATP.

    PubMed

    Matyáš, Robert; Chýlková, Jaromíra

    2013-05-10

    Triacetone triperoxide (3,3,6,6,9,9-hexamethyl-1,2,4,5,7,8-hexoxonane, TATP) is nowadays one of the most commonly used improvised explosives. It is prepared by the action of hydrogen peroxide on acetone in an acidic environment. Easily available mineral acids - hydrochloric, sulfuric, nitric and perchloric - are the most often recommended on the extremist web pages dealing with improvised production of explosives. The various TATP producers' choice of acid mainly depends on the author's experiences and the local availability of the acid. A knowledge of the kind of acid used for TATP production can help in detecting the person who has made the TATP, or who has committed a criminal act using TATP. Therefore, a capillary isotachophoretic method was developed for determination of residual anions (originating from the acid used during TATP synthesis) in the resulting TATP crystals. This analytical method has proved to be reliable; the acid used for TATP synthesis was correctly identified in all samples analyzed. PMID:23542054

  1. Recent developments in detection methods for microfabricated analytical devices.

    PubMed

    Schwarz, M A; Hauser, P C

    2001-09-01

    Sensitive detection in microfluidic analytical devices is a challenge because of the extremely small detection volumes available. Considerable efforts have been made lately to further address this aspect and to investigate techniques other than fluorescence. Among the newly introduced techniques are the optical methods of chemiluminescence, refraction and thermooptics, as well as the electrochemical methods of amperometry, conductimetry and potentiometry. Developments are also in progress to create miniaturized plasma-emission spectrometers and sensitive detectors for gas-chromatographic separations.

  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. 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.

  4. Enhanced Sensitivity for Hydrogen Peroxide Detection: Polydiacetylene Vesicles with Phenylboronic Acid Head Group.

    PubMed

    Jia, Chen; Tang, Jie; Lu, Shengguo; Han, Yuwang; Huang, He

    2016-01-01

    It was recently reported that, besides UV irradiated polymerization, polymerization of diacetylene compounds could also been initiated by radicals generated from enzyme catalyzed hydrogen peroxide (H2O2) decomposition. A new optical sensing method for H2O2 was proposed based on this phenomenon. However, the sensitivity of this method is relatively lower than existed ones. In the present work, phenylboronic acid (PBA) functionalized 10, 12-pentacosadiynoic acid (PDA-PBA) was synthesized and its vesicles were formed successfully as colorimetric sensor for H2O2 detection. It was found that color change during the polymerization of vesicles composed of the PBA modified monomer is much stronger than that of the non-modified one. The response of PDA-PBA vesicles to H2O2 is 16 times more sensitive than that of the PDA. The absorption of PDA-PBA at 650 nm is linearly related to the concentration of H2O2 and a detection limit of ~5 μM could be achieved.

  5. Novel microbial screen for detection of 1,4-butanediol, ethylene glycol, and adipic acid.

    PubMed

    Stieglitz, B; Weimer, P J

    1985-03-01

    A novel microbial-screening procedure was developed for separate detection of 1,4-butanediol, ethylene glycol, and adipic acid, three commercially important oxychemicals potentially derivable from bacterial omega-oxidation of n-butanol, ethanol, and hexanoic acid, respectively. The screening method involved postproduction addition of one of several specific Pseudomonas strains which produce a soluble fluorescent pigment during growth on the product of interest. A mutation and selection procedure was developed for isolation of specific strains with phenotypes for growth and pigment production on the desired product (e.g., 1,4-butanediol), but not on its bioconversion substrate (e.g., n-butanol), common by-products (e.g., n-butyrate), or product isomers. Pigment production was growth associated and required cultivation of the screening strains under limiting Fe3+ concentrations. The pigments resembled well-characterized, iron-chelating siderophores produced by other fluorescent pseudomonads. The sensitivity of the assay for product accumulation was enhanced by (i) conducting the screening in microtiter dishes to permit examination of individual isolates of putative producers and to control product diffusion, (ii) using a wavelength cutoff filter to reduce background source light, and (iii) using adapted screening strains which grew at lower (0.3 mM) concentrations of test compounds. The potential utility of the method for detecting a variety of oxidative catabolic products is discussed.

  6. Determination of Vitamin C (Ascorbic Acid) Using High Performance Liquid Chromatography Coupled with Electrochemical Detection

    PubMed Central

    Gazdik, Zbynek; Zitka, Ondrej; Petrlova, Jitka; Adam, Vojtech; Zehnalek, Josef; Horna, Ales; Reznicek, Vojtech; Beklova, Miroslava; Kizek, Rene

    2008-01-01

    Vitamin C (ascorbic acid, ascorbate, AA) is a water soluble organic compound that participates in many biological processes. The main aim of this paper was to utilize two electrochemical detectors (amperometric – Coulouchem III and coulometric – CoulArray) coupled with flow injection analysis for the detection of ascorbic acid. Primarily, we optimized the experimental conditions. The optimized conditions were as follows: detector potential 100 mV, temperature 25 °C, mobile phase 0.09% TFA:ACN, 3:97 (v/v) and flow rate 0.13 mL·min-1. The tangents of the calibration curves were 0.3788 for the coulometric method and 0.0136 for the amperometric one. The tangent of the calibration curve measured by the coulometric detector was almost 30 times higher than the tangent measured by the amperometric detector. Consequently, we coupled a CoulArray electrochemical detector with high performance liquid chromatography and estimated the detection limit for AA as 90 nM (450 fmol per 5 μL injection). The method was used for the determination of vitamin C in a pharmaceutical preparations (98 ± 2 mg per tablet), in oranges (Citrus aurantium) (varied from 30 to 56 mg/100 g fresh weight), in apples (Malus sp.) (varied from 11 to 19 mg/100 g fresh weight), and in human blood serum (varied from 38 to 78 μM). The recoveries were also determined.

  7. Improved Hidden-Markov-Model Method Of Detecting Faults

    NASA Technical Reports Server (NTRS)

    Smyth, Padhraic J.

    1994-01-01

    Method of automated, continuous monitoring to detect faults in complicated dynamic system based on hidden-Markov-model (HMM) approach. Simpler than another, recently proposed HMM method, but retains advantages of that method, including low susceptibility to false alarms, no need for mathematical model of dynamics of system under normal or faulty conditions, and ability to detect subtle changes in characteristics of monitored signals. Examples of systems monitored by use of this method include motors, turbines, and pumps critical in their applications; chemical-processing plants; powerplants; and biomedical systems.

  8. Chromatographic method for determination of the free amino acid content of chamomile flowers

    PubMed Central

    Ma, Xiaoli; Zhao, Dongsheng; Li, Xinxia; Meng, Lei

    2015-01-01

    Objective: To determine the free amino acid contents of chamomile flowers using reverse-phase high-performance column chromatography preceded by pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC), and to determine the reliability of this method. Materials and Methods: Derivatization with reconstituted AQC was used to prepare the samples and standards for injection into the chromatography column. The peaks were analyzed by fluorescence detection (λ excitation, 250 nm; λ emission, 395 nm. Results: Alanine, proline, and leucine were the most abundant amino acids, whereas tyrosine and methionine were the least abundant. The linearity of the method was found to be good with amino acid concentrations of 0.012-0.36 μM. The precision was 0.05-1.36%; average recovery, 91.12-129.41%; and limit of detection, 0.006-0.058 μM. Conclusion: The method is reliable for determining the free amino acid content of different types of chamomile flowers. PMID:25709230

  9. Method for edge detection in images using fuzzy relation

    NASA Astrophysics Data System (ADS)

    Nieradka, Grzegorz

    2008-01-01

    Edge detection is one of the most important tasks in machine vision and image processing. The paper presents novel approach to construction of algorithm for detection edges in images. In order to solve the problem the concepts of the fuzzy similarity relation and homogeneity region were exploited. The results of application of the proposed edge calculation method are presented in this paper. Additionally, this paper presents an interesting artistic effect obtained by application of the image processing method based on such the detection of edges. Possible artistic effects of application of this algorithm are exemplified.

  10. A Highly Sensitive Kinetic Spectrophotometric Method for the Determination of Ascorbic Acid in Pharmaceutical Samples

    PubMed Central

    Shishehbore, Masoud Reza; Aghamiri, Zahra

    2014-01-01

    In this study, a new reaction system for quantitative determination of ascorbic acid was introduced. The developed method is based on inhibitory effect of ascorbic acid on the Orange G-bromate system. The change in absorbance was followed spectrophotometrically at 478 nm. The dependence of sensitivity on the reaction variables including reagents concentration, temperature and time was investigated. Under optimum experimental conditions, calibration curve was linear over the range 0.7 – 33.5 μg mL-1 of ascorbic acid including two linear segments and the relative standard deviations (n = 6) for 5.0 and 20.0 μg mL–1 of ascorbic acid were 1.08 and 1.02%, respectively. The limit of detection was 0.21 μg mL−1 of ascorbic acid. The effect of diverse species was also investigated. The developed method was successfully applied for the determination of ascorbic acid in pharmaceutical samples. The results were in a good agreement with those of reference method. PMID:25237333

  11. Determination of usnic acid in lichen toxic to elk by liquid chromatography with ultraviolet and tandem mass spectrometry detection.

    PubMed

    Roach, John A G; Musser, Steven M; Morehouse, Kim; Woo, Jason Y J

    2006-04-01

    Usnic acid is unambiguously confirmed by tandem mass spectrometry (MS/MS) in tumbleweed shield lichen, Xanthoparmelia chlorochroa. The lichen contains 2% usnic acid by liquid chromatography with UV quantification at 282 nm. The UV linear range for usnic acid quantification is from its 4 ng limit of detection to 2 microg injected. UV signal saturation is recognized by distortion of the usnic acid UV spectrum. Positive ion electrospray-tandem mass spectrometry offers no similar means to recognize quantification data recorded above the linear range of electrospray. Electrospray ionization capacity and matrix effects limit the reliability of tandem mass spectrometry quantification. The combination of UV quantification and MS confirmation provides a reliable analytical method for measuring usnic acid levels in plant material. PMID:16569032

  12. Determination of usnic acid in lichen toxic to elk by liquid chromatography with ultraviolet and tandem mass spectrometry detection.

    PubMed

    Roach, John A G; Musser, Steven M; Morehouse, Kim; Woo, Jason Y J

    2006-04-01

    Usnic acid is unambiguously confirmed by tandem mass spectrometry (MS/MS) in tumbleweed shield lichen, Xanthoparmelia chlorochroa. The lichen contains 2% usnic acid by liquid chromatography with UV quantification at 282 nm. The UV linear range for usnic acid quantification is from its 4 ng limit of detection to 2 microg injected. UV signal saturation is recognized by distortion of the usnic acid UV spectrum. Positive ion electrospray-tandem mass spectrometry offers no similar means to recognize quantification data recorded above the linear range of electrospray. Electrospray ionization capacity and matrix effects limit the reliability of tandem mass spectrometry quantification. The combination of UV quantification and MS confirmation provides a reliable analytical method for measuring usnic acid levels in plant material.

  13. Other Notable Methods of Membrane Protein Detection: A Brief Review.

    PubMed

    Kurien, Biji T; Scofield, R Hal

    2015-01-01

    Several techniques have been employed to detect proteins on membranes. These include the use of quantum dot luminescent labels, oxyblot immunochemical detection, polymer immunocomplexes, "coupled" probing approach, in situ renaturation of proteins for detecting enzyme activities in crude or purified preparations, immunochromatographic assay, western-phosphatase assay, and use of Congo red dye (a cosmetic color named Alta), Pro-Q Emerald 488 dye, or amine-reactive dye in combination with alkaline phosphatase- and horseradish peroxidase-antibody conjugates for the simultaneous trichromatic fluorescence detection of proteins. Several methods have been used to improve the detection of proteins on membranes, including glutaraldehyde treatment of nitrocellulose blots, elimination of keratin artifacts in immunoblots probed with polyclonal antibodies, and washing of immunoblots with excessive water and manipulation of Tween-20 in wash buffer. These methods are briefly reviewed in this chapter. PMID:26139283

  14. 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.

  15. Immobilization-free electrochemical DNA detection with anthraquinone-labeled pyrrolidinyl peptide nucleic acid probe.

    PubMed

    Kongpeth, Jutatip; Jampasa, Sakda; Chaumpluk, Piyasak; Chailapakul, Orawon; Vilaivan, Tirayut

    2016-01-01

    Electrochemical detection provides a simple, rapid, sensitive and inexpensive method for DNA detection. In traditional electrochemical DNA biosensors, the probe is immobilized onto the electrode. Hybridization with the DNA target causes a change in electrochemical signal, either from the intrinsic signal of the probe/target or through a label or a redox indicator. The major drawback of this approach is the requirement for probe immobilization in a controlled fashion. In this research, we take the advantage of different electrostatic properties between PNA and DNA to develop an immobilization-free approach for highly sequence-specific electrochemical DNA sensing on a screen-printed carbon electrode (SPCE) using a square-wave voltammetric (SWV) technique. Anthraquinone-labeled pyrrolidinyl peptide nucleic acid (AQ-PNA) was employed as a probe together with an SPCE that was modified with a positively-charged polymer (poly quaternized-(dimethylamino-ethyl)methacrylate, PQDMAEMA). The electrostatic attraction between the negatively-charged PNA-DNA duplex and the positively-charged modified SPCE attributes to the higher signal of PNA-DNA duplex than that of the electrostatically neutral PNA probe, resulting in a signal change. The calibration curve of this proposed method exhibited a linear range between 0.35 and 50 nM of DNA target with a limit of detection of 0.13 nM (3SD(blank)/Slope). The sub-nanomolar detection limit together with a small sample volume required (20 μL) allowed detection of <10 fmol (<1 ng) of DNA. With the high specificity of the pyrrolidinyl PNA probe used, excellent discrimination between complementary and various single-mismatched DNA targets was obtained. An application of this new platform for a sensitive and specific detection of isothermally-amplified shrimp's white spot syndrome virus (WSSV) DNA was successfully demonstrated. PMID:26695270

  16. Immobilization-free electrochemical DNA detection with anthraquinone-labeled pyrrolidinyl peptide nucleic acid probe.

    PubMed

    Kongpeth, Jutatip; Jampasa, Sakda; Chaumpluk, Piyasak; Chailapakul, Orawon; Vilaivan, Tirayut

    2016-01-01

    Electrochemical detection provides a simple, rapid, sensitive and inexpensive method for DNA detection. In traditional electrochemical DNA biosensors, the probe is immobilized onto the electrode. Hybridization with the DNA target causes a change in electrochemical signal, either from the intrinsic signal of the probe/target or through a label or a redox indicator. The major drawback of this approach is the requirement for probe immobilization in a controlled fashion. In this research, we take the advantage of different electrostatic properties between PNA and DNA to develop an immobilization-free approach for highly sequence-specific electrochemical DNA sensing on a screen-printed carbon electrode (SPCE) using a square-wave voltammetric (SWV) technique. Anthraquinone-labeled pyrrolidinyl peptide nucleic acid (AQ-PNA) was employed as a probe together with an SPCE that was modified with a positively-charged polymer (poly quaternized-(dimethylamino-ethyl)methacrylate, PQDMAEMA). The electrostatic attraction between the negatively-charged PNA-DNA duplex and the positively-charged modified SPCE attributes to the higher signal of PNA-DNA duplex than that of the electrostatically neutral PNA probe, resulting in a signal change. The calibration curve of this proposed method exhibited a linear range between 0.35 and 50 nM of DNA target with a limit of detection of 0.13 nM (3SD(blank)/Slope). The sub-nanomolar detection limit together with a small sample volume required (20 μL) allowed detection of <10 fmol (<1 ng) of DNA. With the high specificity of the pyrrolidinyl PNA probe used, excellent discrimination between complementary and various single-mismatched DNA targets was obtained. An application of this new platform for a sensitive and specific detection of isothermally-amplified shrimp's white spot syndrome virus (WSSV) DNA was successfully demonstrated.

  17. Improved liquid chromatographic method for determination of organic acids in leaves, pulp, fruits, and rinds of Garcinia.

    PubMed

    Jayaprakasha, Guddadarangavvanahally K; Jena, Bhabani S; Sakariah, Kunnumpurath K

    2003-01-01

    An improved liquid chromatographic (LC) method was developed for determination of organic acids in leaves, pulp, fruits, and rinds of Garcinia. At present, the commonly used LC method for analysis of organic acids in Garcinia extracts uses direct application of the extracts on the column. This practice gradually reduces efficiency of the column and shortens its life. In the improved method, the interfering substances such as pigments and xanthones were effectively removed by passing the aqueous extract through an ODS cartridge. With subsequent injection on a C18 reversed-phase column, using 6.0 mM phosphoric acid as the mobile phase with a flow rate of 1.0 mL/min and UV detection at 210 nm, the organic acids were determined in the extracts. The major organic acid was (-)-hydroxycitric acid at the level of 2.5, 0.8, 3.0, and 20.1% in leaf, pulp, fresh fruit, and dried rinds, respectively. Minor quantities of hydroxycitric acid lactone, oxalic acid, and citric acid were also identified. Limits of detection and recoveries were 0.9-1.5 microg and 93.9-99.8%, respectively. This is the first report on the composition of organic acids from Garcinia pedunculata.

  18. 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.

  19. 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.

  20. Quantitative detection of Aspergillus spp. by real-time nucleic acid sequence-based amplification.

    PubMed

    Zhao, Yanan; Perlin, David S

    2013-01-01

    Rapid and quantitative detection of Aspergillus from clinical samples may facilitate an early diagnosis of invasive pulmonary aspergillosis (IPA). As nucleic acid-based detection is a viable option, we demonstrate that Aspergillus burdens can be rapidly and accurately detected by a novel real-time nucleic acid assay other than qPCR by using the combination of nucleic acid sequence-based amplification (NASBA) and the molecular beacon (MB) technology. Here, we detail a real-time NASBA assay to determine quantitative Aspergillus burdens in lungs and bronchoalveolar lavage (BAL) fluids of rats with experimental IPA.

  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. 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.

  3. 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.

  4. 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.

  5. Ultra-sensitive detection of zinc oxide nanowires using a quartz crystal microbalance and phosphoric acid DNA.

    PubMed

    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.

  6. Ultra-sensitive detection of zinc oxide nanowires using a quartz crystal microbalance and phosphoric acid DNA.

    PubMed

    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. PMID:27479871

  7. 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.

  8. 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

  9. A method for detecting positive growth autocorrelation without marking individuals.

    PubMed

    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 ρ (2) = 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

  10. 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

  11. 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

  12. 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.

  13. Superhydrophilic cotton thread with temperature-dependent pattern for sensitive nucleic acid detection.

    PubMed

    Wu, Tingting; Xu, Tailin; Xu, Li-Ping; Huang, Yan; Shi, Wanxin; Wen, Yongqiang; Zhang, Xueji

    2016-12-15

    Cotton thread is promising in fabricating biosensors for diagnostic application due to its excellent characteristics. However, the enrichment of the capture molecules on a narrow zone of the cotton thread based biosensor is a big challenge because of its superhydrophilicity. Here, we report a simple, low-cost and accurate cotton thread based nucleic acid biosensor with temperature-dependent pattern. Liquid wax is used to fabricate temperature-dependent pattern to restrict the test zone in a narrow area. This biosensor enables visual and quantitative detection of target DNA by accumulation of gold nanoparticles (GNPs) on the test zone with a detection limits of 0.75nM. In addition, the cotton thread based biosensor needs less sample than previous reported lateral flow strip and the sample solution wicks faster at the cotton thread which can lead to a shorter detection time. This simple, low-cost and fast detection method holds great potential to improve healthcare services in the developing regions. PMID:27498320

  14. A Comparison of Satellite-Based Multilayered Cloud Detection Methods

    NASA Technical Reports Server (NTRS)

    Minnis, Patrick; Chang, Fu-Lung; Khaiyer, Mandana M.; Ayers, Jeffrey K.; Palikonda, Rabindra; Nordeen, Michele L.; Spangenberg, Douglas A.

    2007-01-01

    Both techniques show skill in detecting multilayered clouds, but they disagree more than 50% of the time. BTD method tends to detect more ML clouds than CO2 method and has slightly higher detection accuracy. CO2 method might be better for minimizing false positives, but further study is needed. Neither method as been optimized for GOES data. BTD technique developed on AVHRR, better BTD signals & resolution. CO2 developed on MODIS, better resolution & 4 CO2 channels. Many additional comparisons with ARSCL data will be used to optimize both techniques. A combined technique will be examined using MODIS & Meteosat-8 data. After optimization, the techniques will be implemented in the ARM operational satellite cloud processing.

  15. 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

  16. Laser-based optical activity detection of amino acids and proteins

    SciTech Connect

    Reitsma, B.H.

    1987-08-01

    The optical activity detector (OAD) for HPLC is a selective detector for optically active substances including amino acids and proteins. Four free amino acids were resolved using cation-exchange chromatography followed by detection with refractive index detector (RI) for proline and threonine and the OAD to an ultraviolet absorbance detector (uv) for tyrosine and phenylalanine. Amino acid detection by refractive index is not sensitive and uv absorbance detects only three amino acids. Derivatization of amino acids to make them detectable by uv absorbance enhances the applicability of OA/uv for the determination of enantiomeric ratios. The separation of 16 dansyl-L-amino acids by RP-HPLC with detection by OA/uv is illustrated. Calculation of the specific rotation of 22 dansyl-L-amino acids shows that derivatization enhances the OA detectability of some amino acids but degrades that of others. RP-HPLC of proteins is a rapidly developing technique. Several researchers have reported the detection of multiple peaks when a pure protein is subjected to HPLC under certain conditions. These multiple peaks have been determined to be different conformations of the same protein. Since proteins are optically active, OA is a suitable detector. The RP-HPLC separation of conformers of soybean trypsin inhibitor is illustrated. Detection by OA/uv provides insights from the chromatogram unavailable from uv absorbance detection alone. In addition, identification of impurities is simplified with OA/uv. Specific rotations of the separated protein fractions show no significant change accompanying change in conformation. 163 refs., 13 figs., 9 tabs.

  17. Electroactive chitosan nanoparticles for the detection of single-nucleotide polymorphisms using peptide nucleic acids.

    PubMed

    Kerman, Kagan; Saito, Masato; Tamiya, Eiichi

    2008-08-01

    Here we report an electrochemical biosensor that would allow for simple and rapid analysis of nucleic acids in combination with nuclease activity on nucleic acids and electroactive bionanoparticles. The detection of single-nucleotide polymorphisms (SNPs) using PNA probes takes advantage of the significant structural and physicochemical differences between the full hybrids and SNPs in PNA/DNA and DNA/DNA duplexes. Ferrocene-conjugated chitosan nanoparticles (Chi-Fc) were used as the electroactive indicator of hybridization. Chi-Fc had no affinity towards the neutral PNA probe immobilized on a gold electrode (AuE) surface. When the PNA probe on the electrode surface hybridized with a full-complementary target DNA, Chi-Fc electrostatically attached to the negatively-charged phosphate backbone of DNA on the surface and gave rise to a high electrochemical oxidation signal from ferrocene at approximately 0.30 V. Exposing the surface to a single-stranded DNA specific nuclease, Nuclease S1, was found to be very effective for removing the nonspecifically adsorbed SNP DNA. An SNP in the target DNA to PNA made it susceptible to the enzymatic digestion. After the enzymatic digestion and subsequent exposure to Chi-Fc, the presence of SNPs was determined by monitoring the changes in the electrical current response of Chi-Fc. The method provided a detection limit of 1 fM (S/N = 3) for the target DNA oligonucleotide. Additionally, asymmetric PCR was employed to detect the presence of genetically modified organism (GMO) in standard Roundup Ready soybean samples. PNA-mediated PCR amplification of real DNA samples was performed to detect SNPs related to alcohol dehydrogenase (ALDH). Chitosan nanoparticles are promising biomaterials for various analytical and pharmaceutical applications.

  18. Detection of Dengue Viral RNA Using a Nucleic Acid Sequence-Based Amplification Assay

    PubMed Central

    Wu, Shuenn-Jue L.; Lee, Eun Mi; Putvatana, Ravithat; Shurtliff, Roxanne N.; Porter, Kevin R.; Suharyono, Wuryadi; Watts, Douglas M.; King, Chwan-Chuen; Murphy, Gerald S.; Hayes, Curtis G.; Romano, Joseph W.

    2001-01-01

    Faster techniques are needed for the early diagnosis of dengue fever and dengue hemorrhagic fever during the acute viremic phase of infection. An isothermal nucleic acid sequence-based amplification (NASBA) assay was optimized to amplify viral RNA of all four dengue virus serotypes by a set of universal primers and to type the amplified products by serotype-specific capture probes. The NASBA assay involved the use of silica to extract viral nucleic acid, which was amplified without thermocycling. The amplified product was detected by a probe-hybridization method that utilized electrochemiluminescence. Using normal human plasma spiked with dengue viruses, the NASBA assay had a detection threshold of 1 to 10 PFU/ml. The sensitivity and specificity of the assay were determined by testing 67 dengue virus-positive and 21 dengue virus-negative human serum or plasma samples. The “gold standard” used for comparison and evaluation was the mosquito C6/36 cell culture assay followed by an immunofluorescent assay. Viral infectivity titers in test samples were also determined by a direct plaque assay in Vero cells. The NASBA assay was able to detect dengue viral RNA in the clinical samples at plaque titers below 25 PFU/ml (the detection limit of the plaque assay). Of the 67 samples found positive by the C6/36 assay, 66 were found positive by the NASBA assay, for a sensitivity of 98.5%. The NASBA assay had a specificity of 100% based on the negative test results for the 21 normal human serum or plasma samples. These results indicate that the NASBA assay is a promising assay for the early diagnosis of dengue infections. PMID:11473994

  19. DNA Diagnostics: Nanotechnology-enhanced Electrochemical Detection of Nucleic Acids

    PubMed Central

    Wei, Fang; Lillehoj, Peter B.; Ho, Chih-Ming

    2010-01-01

    The detection of mismatched base pairs in DNA plays a crucial role in the diagnosis of genetic-related diseases and conditions, especially for early stage treatment. Among the various biosensors that have been employed for DNA detection, electrochemical sensors show great promise since they are capable of precise DNA recognition and efficient signal transduction. Advancements in micro- and nanotechnologies, specifically fabrication techniques and new nanomaterials, have enabled for the development of highly sensitive, highly specific sensors making them attractive for the detection of small sequence variations. Furthermore, the integration of sensors with sample preparation and fluidic processes enables for rapid, multiplexed DNA detection for point-of-care (POC) clinical diagnostics. PMID:20075759

  20. On the detectability of acid formation in clouds

    SciTech Connect

    Kelly, T.J.; Schwartz, S.E.; Daum, P.H. )

    1987-01-01

    Deposition of acids from the atmosphere to the earth's surface in hydrometeors (precipitation or impacted fog droplets) is a well-documented phenomenon. The acids involved are primarily sulfuric (H{sub 2}SO{sub 4}) and nitric (HNO{sub 3}) acids, derived from sulfur dioxide (SO{sub 2}) and oxides of nitrogen (NO and NO{sub 2}, collectively called NO{sub x}). This so-called wet deposition is an important means of removing sulfur- and nitrogen-containing pollutants from the atmosphere, but is also considered a serious environmental problem because of the potential harmful effects of the deposited materials on vegetation and aquatic life. The chemical composition of precipitation is determined primarily by the composition of cloudwater in the clouds from which the precipitation falls. Two processes which determine the cloudwater composition can be distinguised: dissolution of soluble aerosol and gaseous species at the time of cloud formation, and production of acids in the cloud from gaseous SO{sub 2} and NO{sub x}. These processes are discussed by the authors.

  1. Transistor-based particle detection systems and methods

    DOEpatents

    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.

  2. 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.

  3. 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.

  4. Detecting diversity: emerging methods to estimate species diversity.

    PubMed

    Iknayan, Kelly J; Tingley, Morgan W; Furnas, Brett J; Beissinger, Steven R

    2014-02-01

    Estimates of species richness and diversity are central to community and macroecology and are frequently used in conservation planning. Commonly used diversity metrics account for undetected species primarily by controlling for sampling effort. Yet the probability of detecting an individual can vary among species, observers, survey methods, and sites. We review emerging methods to estimate alpha, beta, gamma, and metacommunity diversity through hierarchical multispecies occupancy models (MSOMs) and multispecies abundance models (MSAMs) that explicitly incorporate observation error in the detection process for species or individuals. We examine advantages, limitations, and assumptions of these detection-based hierarchical models for estimating species diversity. Accounting for imperfect detection using these approaches has influenced conclusions of comparative community studies and creates new opportunities for testing theory. PMID:24315534

  5. Visceral perception versus visceral detection: disentangling methods and assumptions.

    PubMed

    Pennebaker, J W; Hoover, C W

    1984-09-01

    A within-subject experiment compared three paradigms commonly used in visceral perception: self-report, heartbeat tracking, and signal detection. Eighteen undergraduates estimated heart rate using each technique while engaging in a number of separate tasks conducted a week apart. Although all three techniques significantly tapped accuracy of heart rate perception, only the self-report and signal detection methods were reliable over time. Most important, there was no relationship involving any of the methods in measuring accuracy. The findings suggest some fundamental differences in the assumptions and perceptual properties of the various paradigms. A distinction is made between visceral perception and detection. Perception implies the subject's use of both internal physiological and external environmental information in the perception of visceral state. Detection connotes the subject's use of only physiological information--to the exclusion of all other factors. The relevance of these approaches for biofeedback and real-world symptom perception is discussed.

  6. 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

  7. 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.

  8. 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.

  9. Laboratory Detection and Analysis of Organic Compounds in Rocks Using HPLC and XRD Methods

    NASA Technical Reports Server (NTRS)

    Dragoi, D.; Kanik, I.; Bar-Cohen, Y.; Sherrit, S.; Tsapin, A.; Kulleck, J.

    2004-01-01

    In this work we describe an analytical method for determining the presence of organic compounds in rocks, limestone, and other composite materials. Our preliminary laboratory experiments on different rocks/limestone show that the organic component in mineralogical matrices is a minor phase on order of hundreds of ppm and can be better detected using high precision liquid chromatography (HPLC). The matrix, which is the major phase, plays an important role in embedding and protecting the organic molecules from the harsh Martian environment. Some rocks bear significant amounts of amino acids therefore, it is possible to identify these phases using powder x-ray diffraction (XRD) by crystallizing the organic. The method of detection/analysis of organics, in particular amino acids, that have been associated with life will be shown in the next section.

  10. Reverse Transcription Cross-Priming Amplification–Nucleic Acid Test Strip for Rapid Detection of Porcine Epidemic Diarrhea Virus

    PubMed Central

    Wang, Feng-Xue; Yuan, Dan-Yi; Jin, Ya-Nan; Hu, Lin; Sun, Zhi-Yong; He, Qian; Zhao, Shi-Hua; Zhan, Shu-Bai; Wen, Yong-Jun

    2016-01-01

    Porcine epidemic diarrhea virus (PEDV) is a highly transmissible coronavirus that causes a severe enteric disease particularly in neonatal piglets. In this study, a rapid method for detecting PEDV was developed based on cross-priming amplification and nucleic acid test strip(CPA-NATS). Five primers specific for the N gene sequence of PEDV were used for the cross-priming amplification. Detection of amplification products based on labeled probe primers was conducted with strip binding antibody of labeled markers. The CPA method was evaluated and compared with a PCR method. The reverse transcription CPA system was further optimized for detecting PEDV RNA in clinical specimens. Results showed that the method was highly specific for the detection of PEDV, and had the same sensitivity as PCR, with detection limit of 10−6 diluted plasmid containing the target gene of PEDV. It was also successfully applied to detecting PEDV in clinical specimens. The reverse transcription CPA-NATS detection system established in this study offers a specific, sensitive, rapid, and simple detection tool for screening PEDV, which can contribute to strategies in the effective control of PEDV in swine. PMID:27090105

  11. CRISPR Spacer Arrays for Detection of Viral Signatures from Acidic Hot Springs

    NASA Astrophysics Data System (ADS)

    Snyder, J. C.; Bateson, M. M.; Suciu, D.; Young, M. J.

    2010-04-01

    Viruses are the most abundant life-like entities on the planet Earth. Using CRISPR spacer sequences, we have developed a microarray-based approach to detecting viral signatures in the acidic hot springs of Yellowstone.

  12. A room-temperature adenosine-based molecular beacon for highly sensitive detection of nucleic acids.

    PubMed

    Lin, Yen-Hsiu; Tseng, Wei-Lung

    2012-06-25

    This study developed a simple, sensitive, and selective molecular beacon for detecting nucleic acids at room temperature based on coralyne induced conformational change of a MB through A(2)-coralyne-A(2) coordination.

  13. Maximum patch method for directional dark matter detection

    SciTech Connect

    Henderson, Shawn; Monroe, Jocelyn; Fisher, Peter

    2008-07-01

    Present and planned dark matter detection experiments search for WIMP-induced nuclear recoils in poorly known background conditions. In this environment, the maximum gap statistical method provides a way of setting more sensitive cross section upper limits by incorporating known signal information. We give a recipe for the numerical calculation of upper limits for planned directional dark matter detection experiments, that will measure both recoil energy and angle, based on the gaps between events in two-dimensional phase space.

  14. Safety assessment and detection methods of genetically modified organisms.

    PubMed

    Xu, Rong; Zheng, Zhe; Jiao, Guanglian

    2014-01-01

    Genetically modified organisms (GMOs), are gaining importance in agriculture as well as the production of food and feed. Along with the development of GMOs, health and food safety concerns have been raised. These concerns for these new GMOs make it necessary to set up strict system on food safety assessment of GMOs. The food safety assessment of GMOs, current development status of safety and precise transgenic technologies and GMOs detection have been discussed in this review. The recent patents about GMOs and their detection methods are also reviewed. This review can provide elementary introduction on how to assess and detect GMOs. PMID:25342147

  15. A generic nuclei detection method for histopathological breast images

    NASA Astrophysics Data System (ADS)

    Kost, Henning; Homeyer, André; Bult, Peter; Balkenhol, Maschenka C. A.; van der Laak, Jeroen A. W. M.; Hahn, Horst K.

    2016-03-01

    The detection of cell nuclei plays a key role in various histopathological image analysis problems. Considering the high variability of its applications, we propose a novel generic and trainable detection approach. Adaption to specific nuclei detection tasks is done by providing training samples. A trainable deconvolution and classification algorithm is used to generate a probability map indicating the presence of a nucleus. The map is processed by an extended watershed segmentation step to identify the nuclei positions. We have tested our method on data sets with different stains and target nuclear types. We obtained F1-measures between 0.83 and 0.93.

  16. Method and automated apparatus for detecting coliform organisms

    NASA Technical Reports Server (NTRS)

    Dill, W. P.; Taylor, R. E.; Jeffers, E. L. (Inventor)

    1980-01-01

    Method and automated apparatus are disclosed for determining the time of detection of metabolically produced hydrogen by coliform bacteria cultured in an electroanalytical cell from the time the cell is inoculated with the bacteria. The detection time data provides bacteria concentration values. The apparatus is sequenced and controlled by a digital computer to discharge a spent sample, clean and sterilize the culture cell, provide a bacteria nutrient into the cell, control the temperature of the nutrient, inoculate the nutrient with a bacteria sample, measures the electrical potential difference produced by the cell, and measures the time of detection from inoculation.

  17. Safety assessment and detection methods of genetically modified organisms.

    PubMed

    Xu, Rong; Zheng, Zhe; Jiao, Guanglian

    2014-01-01

    Genetically modified organisms (GMOs), are gaining importance in agriculture as well as the production of food and feed. Along with the development of GMOs, health and food safety concerns have been raised. These concerns for these new GMOs make it necessary to set up strict system on food safety assessment of GMOs. The food safety assessment of GMOs, current development status of safety and precise transgenic technologies and GMOs detection have been discussed in this review. The recent patents about GMOs and their detection methods are also reviewed. This review can provide elementary introduction on how to assess and detect GMOs.

  18. Machine Learning Methods for Attack Detection in the Smart Grid.

    PubMed

    Ozay, Mete; Esnaola, Inaki; Yarman Vural, Fatos Tunay; Kulkarni, Sanjeev R; Poor, H Vincent

    2016-08-01

    Attack detection problems in the smart grid are posed as statistical learning problems for different attack scenarios in which the measurements are observed in batch or online settings. In this approach, machine learning algorithms are used to classify measurements as being either secure or attacked. An attack detection framework is provided to exploit any available prior knowledge about the system and surmount constraints arising from the sparse structure of the problem in the proposed approach. Well-known batch and online learning algorithms (supervised and semisupervised) are employed with decision- and feature-level fusion to model the attack detection problem. The relationships between statistical and geometric properties of attack vectors employed in the attack scenarios and learning algorithms are analyzed to detect unobservable attacks using statistical learning methods. The proposed algorithms are examined on various IEEE test systems. Experimental analyses show that machine learning algorithms can detect attacks with performances higher than attack detection algorithms that employ state vector estimation methods in the proposed attack detection framework.

  19. Machine Learning Methods for Attack Detection in the Smart Grid.

    PubMed

    Ozay, Mete; Esnaola, Inaki; Yarman Vural, Fatos Tunay; Kulkarni, Sanjeev R; Poor, H Vincent

    2016-08-01

    Attack detection problems in the smart grid are posed as statistical learning problems for different attack scenarios in which the measurements are observed in batch or online settings. In this approach, machine learning algorithms are used to classify measurements as being either secure or attacked. An attack detection framework is provided to exploit any available prior knowledge about the system and surmount constraints arising from the sparse structure of the problem in the proposed approach. Well-known batch and online learning algorithms (supervised and semisupervised) are employed with decision- and feature-level fusion to model the attack detection problem. The relationships between statistical and geometric properties of attack vectors employed in the attack scenarios and learning algorithms are analyzed to detect unobservable attacks using statistical learning methods. The proposed algorithms are examined on various IEEE test systems. Experimental analyses show that machine learning algorithms can detect attacks with performances higher than attack detection algorithms that employ state vector estimation methods in the proposed attack detection framework. PMID:25807571

  20. An adaptive high and low impedance fault detection method

    SciTech Connect

    Yu, D.C. ); Khan, S.H. )

    1994-10-01

    An integrated high impedance fault (HIF) and low impedance fault (LIF) detection method is proposed in this paper. For a HIF detection, the proposed technique is based on a number of characteristics of the HIF current. These characteristics are: fault current magnitude, magnitude of the 3rd harmonic current, magnitude of the 5th harmonic current, the angle of the third harmonic current, the angle difference between the third harmonics current and the fundamental voltage, negative sequence current of HIF. These characteristics are identified by modeling the distribution feeders in EMTP. Apart from these characteristics, the above ambient (average) negative sequence current is also considered. An adjustable block out region around the average load current is provided. The average load current is calculated at every 18,000 cycles (5 minutes) interval. This adaptive feature will not only make the proposed scheme more sensitive to the low fault current, but it will also prevent the relay from tripping during the normal load current. In this paper, the logic circuit required for implementing the proposed HIF detection methods is also included. With minimal modifications, the logic developed for the HIF detection can be applied for the low impedance fault (LIF) detection. A complete logic circuit which detects both the HIF and LIF is proposed. Using this combined logic, the need of installing separate devices for HIF and LIF detection can be eliminated.