Sample records for beta tagging application

  1. Atomic force microscopy of ionotropic receptors bearing subunit-specific tags provides a method for determining receptor architecture

    NASA Astrophysics Data System (ADS)

    Neish, Calum S.; Martin, Ian L.; Davies, Martin; Henderson, Robert M.; Edwardson, J. Michael

    2003-08-01

    We have developed an atomic force microscopy (AFM)-based method for the determination of the subunit architecture of ionotropic receptors, and tested the method using the GABAA receptor as a model system. The most common form of the GABAA receptor probably consists of 2alpha1-, 2beta2- and 1gamma2-subunits. We show here that the arrangement of subunits around the central Cl- ion channel can be deduced by AFM of receptors tagged with subunit-specific antibodies. Transfection of cells with DNA encoding alpha1-, beta2- and gamma2-subunits resulted in the production of receptors containing all three subunits, as judged by both immunoblot analysis and the binding of [3H]-Ro15-1788, a specific radioligand for the GABAA receptor. A His6-tag on the alpha1-subunit was used to purify the receptor from membrane fractions of transfected cells. After incubation with anti-His6 immunoglobulin G, some receptors became tagged with either one or two antibody molecules. AFM analysis of complexes containing two bound antibodies showed that the most common angle between the two tags was 135°, close to the value of 144° expected if the two alpha-subunits are separated by a third subunit. This method is applicable to the complete elucidation of the subunit arrangement around the GABAA receptor rosette, and can also be applied to other ionotropic receptors.

  2. Interaction with beta-arrestin determines the difference in internalization behavor between beta1- and beta2-adrenergic receptors.

    PubMed

    Shiina, T; Kawasaki, A; Nagao, T; Kurose, H

    2000-09-15

    The beta(1)-adrenergic receptor (beta(1)AR) shows the resistance to agonist-induced internalization. As beta-arrestin is important for internalization, we examine the interaction of beta-arrestin with beta(1)AR with three different methods: intracellular trafficking of beta-arrestin, binding of in vitro translated beta-arrestin to intracellular domains of beta(1)- and beta(2)ARs, and inhibition of betaAR-stimulated adenylyl cyclase activities by beta-arrestin. The green fluorescent protein-tagged beta-arrestin 2 translocates to and stays at the plasma membrane by beta(2)AR stimulation. Although green fluorescent protein-tagged beta-arrestin 2 also translocates to the plasma membrane, it returns to the cytoplasm 10-30 min after beta(1)AR stimulation. The binding of in vitro translated beta-arrestin 1 and beta-arrestin 2 to the third intracellular loop and the carboxyl tail of beta(1)AR is lower than that of beta(2)AR. The fusion protein of beta-arrestin 1 with glutathione S-transferase inhibits the beta(1)- and beta(2)AR-stimulated adenylyl cyclase activities, although inhibition of the beta(1)AR-stimulated activity requires a higher concentration of the fusion protein than that of the beta(2)AR-stimulated activity. These results suggest that weak interaction of beta(1)AR with beta-arrestins explains the resistance to agonist-induced internalization. This is further supported by the finding that beta-arrestin can induce internalization of beta(1)AR when beta-arrestin 1 does not dissociate from beta(1)AR by fusing to the carboxyl tail of beta(1)AR.

  3. Association study of IL10, IL1beta, and IL1RN and schizophrenia using tag SNPs from a comprehensive database: suggestive association with rs16944 at IL1beta.

    PubMed

    Shirts, Brian H; Wood, Joel; Yolken, Robert H; Nimgaonkar, Vishwajit L

    2006-12-01

    Genetic association studies of several candidate cytokine genes have been motivated by evidence of immune dysfunction among patients with schizophrenia. Intriguing but inconsistent associations have been reported with polymorphisms of three positional candidate genes, namely IL1beta, IL1RN, and IL10. We used comprehensive sequencing data from the Seattle SNPs database to select tag SNPs that represent all common polymorphisms in the Caucasian population at these loci. Associations with 28 tag SNPs were evaluated in 478 cases and 501 unscreened control individuals, while accounting for population sub-structure using the genomic control method. The samples were also stratified by gender, diagnostic category, and exposure to infectious agents. Significant association was not detected after correcting for multiple comparisons. However, meta-analysis of our data combined with previously published association studies of rs16944 (IL1beta -511) suggests that the C allele confers modest risk for schizophrenia among individuals reporting Caucasian ancestry, but not Asians (Caucasians, n=819 cases, 1292 controls; p=0.0013, OR=1.24, 95% CI 1.09, 1.41).

  4. Analysis of archaeological triacylglycerols by high resolution nanoESI, FT-ICR MS and IRMPD MS/MS: Application to 5th century BC-4th century AD oil lamps from Olbia (Ukraine)

    NASA Astrophysics Data System (ADS)

    Garnier, Nicolas; Rolando, Christian; Høtje, Jakob Munk; Tokarski, Caroline

    2009-07-01

    This work presents the precise identification of triacylglycerols (TAGs) extracted from archaeological samples using a methodology based on nanoelectrospray and Fourier transform mass spectrometry. The archaeological TAG identification needs adapted sample preparation protocols to trace samples in advanced degradation state. More precisely, the proposed preparation procedure includes extraction of the lipid components from finely grinded ceramic using dichloromethane/methanol mixture with additional ultrasonication treatment, and TAG purification by solid phase extraction on a diol cartridge. Focusing on the analytical approach, the implementation of "in-house" species-dependent TAG database was investigated using MS and InfraRed Multiphoton Dissociation (IRMPD) MS/MS spectra; several vegetal oils, dairy products and animal fats were studied. The high mass accuracy of the Fourier transform analyzer ([Delta]m below 2.5 ppm) provides easier data interpretation, and allows distinction between products of different origins. In details, the IRMPD spectra of the lithiated TAGs reveal fragmentation reactions including loss of free neutral fatty acid and loss of fatty acid as [alpha],[beta]-unsaturated moieties. Based on the developed preparation procedure and on the constituted database, TAG extracts from 5th century BC to 4th century AD Olbia lamps were analyzed. The structural information obtained succeeds in identifying that bovine/ovine fats were used as fuel used in these archaeological Olbia lamps.

  5. Systematic gene tagging using CRISPR/Cas9 in human stem cells to illuminate cell organization

    PubMed Central

    Roberts, Brock; Haupt, Amanda; Tucker, Andrew; Grancharova, Tanya; Arakaki, Joy; Fuqua, Margaret A.; Nelson, Angelique; Hookway, Caroline; Ludmann, Susan A.; Mueller, Irina A.; Yang, Ruian; Horwitz, Rick; Rafelski, Susanne M.; Gunawardane, Ruwanthi N.

    2017-01-01

    We present a CRISPR/Cas9 genome-editing strategy to systematically tag endogenous proteins with fluorescent tags in human induced pluripotent stem cells (hiPSC). To date, we have generated multiple hiPSC lines with monoallelic green fluorescent protein tags labeling 10 proteins representing major cellular structures. The tagged proteins include alpha tubulin, beta actin, desmoplakin, fibrillarin, nuclear lamin B1, nonmuscle myosin heavy chain IIB, paxillin, Sec61 beta, tight junction protein ZO1, and Tom20. Our genome-editing methodology using Cas9/crRNA ribonuclear protein and donor plasmid coelectroporation, followed by fluorescence-based enrichment of edited cells, typically resulted in <0.1–4% homology-directed repair (HDR). Twenty-five percent of clones generated from each edited population were precisely edited. Furthermore, 92% (36/39) of expanded clonal lines displayed robust morphology, genomic stability, expression and localization of the tagged protein to the appropriate subcellular structure, pluripotency-marker expression, and multilineage differentiation. It is our conclusion that, if cell lines are confirmed to harbor an appropriate gene edit, pluripotency, differentiation potential, and genomic stability are typically maintained during the clonal line–generation process. The data described here reveal general trends that emerged from this systematic gene-tagging approach. Final clonal lines corresponding to each of the 10 cellular structures are now available to the research community. PMID:28814507

  6. Immobilisation of enzymes on poly(aniline)-poly(anion) composite films. Preparation of bioanodes for biofuel cell applications.

    PubMed

    Simon, Evelyne; Halliwell, Catherine M; Toh, Chee Seng; Cass, Anthony E G; Bartlett, Philip N

    2002-01-01

    Immobilisation of enzymes is important for applications such as biosensors or biofuel cells. A poly(histidine) tag had been introduced on the C terminus of a lactate dehydrogenase enzyme. This mutant enzyme was then immobilised onto poly(aniline) (PANi)-poly(anion) composite films, PANi-poly(vinylsulfonate) (PVS) or PANi-poly(acrylate) (PAA). The NADH produced by the immobilised enzyme in the presence of beta-nicotinamide adenine dinucleotide (NAD(+)) and lactate is oxidised at the poly(aniline)-coated electrode at 0.05 to 0.1 V vs. saturated calomel electrode (SCE) at 35 degrees C.

  7. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pirastu, M.; Galanello, R.; Doherty, M.A.

    The predominant ..beta..-thalassemia in Sardinia is the ..beta../sup 0/ type in which no ..beta..-globin chains are synthesized in the homozygous state. The authors determined the ..beta..-thalassemia mutations in this population by the oligonucleotide-probe method and defined the chromosome haplotypes on which the mutation resides. The same ..beta../sup 39(CAG..-->..TAG)/ nonsense mutation was found on nine different chromosome haplotypes. Although this mutation may have arisen more than once, the multiple haplotypes could also be generated by crossing over and gene conversion events. These findings underscore the frequency of mutational events in the ..beta..-globin gene region.

  8. Systematic gene tagging using CRISPR/Cas9 in human stem cells to illuminate cell organization.

    PubMed

    Roberts, Brock; Haupt, Amanda; Tucker, Andrew; Grancharova, Tanya; Arakaki, Joy; Fuqua, Margaret A; Nelson, Angelique; Hookway, Caroline; Ludmann, Susan A; Mueller, Irina A; Yang, Ruian; Horwitz, Rick; Rafelski, Susanne M; Gunawardane, Ruwanthi N

    2017-10-15

    We present a CRISPR/Cas9 genome-editing strategy to systematically tag endogenous proteins with fluorescent tags in human induced pluripotent stem cells (hiPSC). To date, we have generated multiple hiPSC lines with monoallelic green fluorescent protein tags labeling 10 proteins representing major cellular structures. The tagged proteins include alpha tubulin, beta actin, desmoplakin, fibrillarin, nuclear lamin B1, nonmuscle myosin heavy chain IIB, paxillin, Sec61 beta, tight junction protein ZO1, and Tom20. Our genome-editing methodology using Cas9/crRNA ribonuclear protein and donor plasmid coelectroporation, followed by fluorescence-based enrichment of edited cells, typically resulted in <0.1-4% homology-directed repair (HDR). Twenty-five percent of clones generated from each edited population were precisely edited. Furthermore, 92% (36/39) of expanded clonal lines displayed robust morphology, genomic stability, expression and localization of the tagged protein to the appropriate subcellular structure, pluripotency-marker expression, and multilineage differentiation. It is our conclusion that, if cell lines are confirmed to harbor an appropriate gene edit, pluripotency, differentiation potential, and genomic stability are typically maintained during the clonal line-generation process. The data described here reveal general trends that emerged from this systematic gene-tagging approach. Final clonal lines corresponding to each of the 10 cellular structures are now available to the research community. © 2017 Roberts, Haupt, et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  9. Structural characterization of acylimine-containing blue and red chromophores in mTagBFP and TagRFP fluorescent proteins.

    PubMed

    Subach, Oksana M; Malashkevich, Vladimir N; Zencheck, Wendy D; Morozova, Kateryna S; Piatkevich, Kiryl D; Almo, Steven C; Verkhusha, Vladislav V

    2010-04-23

    We determined the 2.2 A crystal structures of the red fluorescent protein TagRFP and its derivative, the blue fluorescent protein mTagBFP. The crystallographic analysis is consistent with a model in which TagRFP has the trans coplanar anionic chromophore with the conjugated pi-electron system, similar to that of DsRed-like chromophores. Refined conformation of mTagBFP suggests the presence of an N-acylimine functionality in its chromophore and single C(alpha)-C(beta) bond in the Tyr64 side chain. Mass spectrum of mTagBFP chromophore-bearing peptide indicates a loss of 20 Da upon maturation, whereas tandem mass spectrometry reveals that the C(alpha)-N bond in Leu63 is oxidized. These data indicate that mTagBFP has a new type of the chromophore, N-[(5-hydroxy-1H-imidazole-2-yl)methylidene]acetamide. We propose a chemical mechanism in which the DsRed-like chromophore is formed via the mTagBFP-like blue intermediate. (c) 2010 Elsevier Ltd. All rights reserved.

  10. Pancreatic Beta Cells Synthesize Neuropeptide Y and Can Rapidly Release Peptide Co-Transmitters

    PubMed Central

    Whim, Matthew D.

    2011-01-01

    Background In addition to polypeptide hormones, pancreatic endocrine cells synthesize a variety of bioactive molecules including classical transmitters and neuropeptides. While these co-transmitters are thought to play a role in regulating hormone release little is known about how their secretion is regulated. Here I investigate the synthesis and release of neuropeptide Y from pancreatic beta cells. Methodology/Principal Findings NPY appears to be an authentic co-transmitter in neonatal, but not adult, beta cells because (1) early in mouse post-natal development, many beta cells are NPY-immunoreactive whereas no staining is observed in beta cells from NPY knockout mice; (2) GFP-expressing islet cells from an NPY(GFP) transgenic mouse are insulin-ir; (3) single cell RT-PCR experiments confirm that the NPY(GFP) cells contain insulin mRNA, a marker of beta cells. The NPY-immunoreactivity previously reported in alpha and delta cells is therefore likely to be due to the presence of NPY-related peptides. INS-1 cells, a beta cell line, are also NPY-ir and contain NPY mRNA. Using the FMRFamide tagging technique, NPY secretion was monitored from INS-1 beta cells with high temporal resolution. Peptide release was evoked by brief depolarizations and was potentiated by activators of adenylate cyclase and protein kinase A. Following a transient depolarization, NPY-containing dense core granules fused with the cell membrane and discharged their contents within a few milliseconds. Conclusions These results indicate that after birth, NPY expression in pancreatic islets is restricted to neonatal beta cells. The presence of NPY suggests that peptide co-transmitters could mediate rapid paracrine or autocrine signaling within the endocrine pancreas. The FMRFamide tagging technique may be useful in studying the release of other putative islet co-transmitters in real time. PMID:21559341

  11. The Enriched Xenon Observatory: EXO-200 and Ba+ tagging

    NASA Astrophysics Data System (ADS)

    Dolinski, M. J.; EXO Collaboration

    2012-08-01

    The Enriched Xenon Observatory (EXO) is a proposed ton-scale double beta decay experiment with a tentative design sensitivity to the Majorana mass of ˜10 meV. The first phase of EXO is EXO-200, which uses 200 kg of Xe enriched to 80% in 136Xe to search for neutrinoless double beta decay. EXO-200 is a liquid Xe time projection chamber with the ability to detect both scintillation and ionization signals. The detector is constructed from ultra-low background materials and is currently installed at the Waste Isolation Pilot Plant, a salt mine with a 1600 meter water equivalent overburden. The projected 2 year sensitivity for EXO-200 is T1/20ν>6.4×1025 y at 90% confidence level. Looking toward a ton-scale EXO, one unique feature of the experiment is the proposal to identify the barium daughter produced by 136Xe double beta decay on an event-by-event basis. This technique will allow for the elimination of all backgrounds other than the background from the two-neutrino double beta decay spectrum. The EXO Collaboration is exploring a number of options to implement Ba-daughter tagging in the next generation EXO experiment.

  12. Behavioral tagging of extinction learning.

    PubMed

    de Carvalho Myskiw, Jociane; Benetti, Fernando; Izquierdo, Iván

    2013-01-15

    Extinction of contextual fear in rats is enhanced by exposure to a novel environment at 1-2 h before or 1 h after extinction training. This effect is antagonized by administration of protein synthesis inhibitors anisomycin and rapamycin into the hippocampus, but not into the amygdala, immediately after either novelty or extinction training, as well as by the gene expression blocker 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole administered after novelty training, but not after extinction training. Thus, this effect can be attributed to a mechanism similar to synaptic tagging, through which long-term potentiation can be enhanced by other long-term potentiations or by exposure to a novel environment in a protein synthesis-dependent fashion. Extinction learning produces a tag at the appropriate synapses, whereas novelty learning causes the synthesis of plasticity-related proteins that are captured by the tag, strengthening the synapses that generated this tag.

  13. First flavor-tagged determination of bounds on mixing-induced CP violation in Bs0 --> J/psiphi decays.

    PubMed

    Aaltonen, T; Adelman, J; Akimoto, T; Albrow, M G; Alvarez González, B; Amerio, S; Amidei, D; Anastassov, A; Annovi, A; Antos, J; Aoki, M; Apollinari, G; Apresyan, A; Arisawa, T; Artikov, A; Ashmanskas, W; Attal, A; Aurisano, A; Azfar, F; Azzi-Bacchetta, P; Azzurri, P; Bacchetta, N; Badgett, W; Barbaro-Galtieri, A; Barnes, V E; Barnett, B A; Baroiant, S; Bartsch, V; Bauer, G; Beauchemin, P-H; Bedeschi, F; Bednar, P; Behari, S; Bellettini, G; Bellinger, J; Belloni, A; Benjamin, D; Beretvas, A; Beringer, J; Berry, T; Bhatti, A; Binkley, M; Bisello, D; Bizjak, I; Blair, R E; Blocker, C; Blumenfeld, B; Bocci, A; Bodek, A; Boisvert, V; Bolla, G; Bolshov, A; Bortoletto, D; Boudreau, J; Boveia, A; Brau, B; Bridgeman, A; Brigliadori, L; Bromberg, C; Brubaker, E; Budagov, J; Budd, H S; Budd, S; Burkett, K; Busetto, G; Bussey, P; Buzatu, A; Byrum, K L; Cabrera, S; Campanelli, M; Campbell, M; Canelli, F; Canepa, A; Carlsmith, D; Carosi, R; Carrillo, S; Carron, S; Casal, B; Casarsa, M; Castro, A; Catastini, P; Cauz, D; Cavalli-Sforza, M; Cerri, A; Cerrito, L; Chang, S H; Chen, Y C; Chertok, M; Chiarelli, G; Chlachidze, G; Chlebana, F; Cho, K; Chokheli, D; Chou, J P; Choudalakis, G; Chuang, S H; Chung, K; Chung, W H; Chung, Y S; Ciobanu, C I; Ciocci, M A; Clark, A; Clark, D; Compostella, G; Convery, M E; Conway, J; Cooper, B; Copic, K; Cordelli, M; Cortiana, G; Crescioli, F; Cuenca Almenar, C; Cuevas, J; Culbertson, R; Cully, J C; Dagenhart, D; Datta, M; Davies, T; de Barbaro, P; De Cecco, S; Deisher, A; De Lentdecker, G; De Lorenzo, G; Dell'Orso, M; Demortier, L; Deng, J; Deninno, M; De Pedis, D; Derwent, P F; Di Giovanni, G P; Dionisi, C; Di Ruzza, B; Dittmann, J R; D'Onofrio, M; Donati, S; Dong, P; Donini, J; Dorigo, T; Dube, S; Efron, J; Erbacher, R; Errede, D; Errede, S; Eusebi, R; Fang, H C; Farrington, S; Fedorko, W T; Feild, R G; Feindt, M; Fernandez, J P; Ferrazza, C; Field, R; Flanagan, G; Forrest, R; Forrester, S; Franklin, M; Freeman, J C; Furic, I; Gallinaro, M; Galyardt, J; Garberson, F; Garcia, J E; Garfinkel, A F; Genser, K; Gerberich, H; Gerdes, D; Giagu, S; Giakoumopolou, V; Giannetti, P; Gibson, K; Gimmell, J L; Ginsburg, C M; Giokaris, N; Giordani, M; Giromini, P; Giunta, M; Giurgiu, G; Glagolev, V; Glenzinski, D; Gold, M; Goldschmidt, N; Golossanov, A; Gomez, G; Gomez-Ceballos, G; Goncharov, M; González, O; Gorelov, I; Goshaw, A T; Goulianos, K; Gresele, A; Grinstein, S; Grosso-Pilcher, C; Grundler, U; Guimaraes da Costa, J; Gunay-Unalan, Z; Haber, C; Hahn, K; Hahn, S R; Halkiadakis, E; Hamilton, A; Han, B-Y; Han, J Y; Handler, R; Happacher, F; Hara, K; Hare, D; Hare, M; Harper, S; Harr, R F; Harris, R M; Hartz, M; Hatakeyama, K; Hauser, J; Hays, C; Heck, M; Heijboer, A; Heinemann, B; Heinrich, J; Henderson, C; Herndon, M; Heuser, J; Hewamanage, S; Hidas, D; Hill, C S; Hirschbuehl, D; Hocker, A; Hou, S; Houlden, M; Hsu, S-C; Huffman, B T; Hughes, R E; Husemann, U; Huston, J; Incandela, J; Introzzi, G; Iori, M; Ivanov, A; Iyutin, B; James, E; Jayatilaka, B; Jeans, D; Jeon, E J; Jindariani, S; Johnson, W; Jones, M; Joo, K K; Jun, S Y; Jung, J E; Junk, T R; Kamon, T; Kar, D; Karchin, P E; Kato, Y; Kephart, R; Kerzel, U; Khotilovich, V; Kilminster, B; Kim, D H; Kim, H S; Kim, J E; Kim, M J; Kim, S B; Kim, S H; Kim, Y K; Kimura, N; Kirsch, L; Klimenko, S; Klute, M; Knuteson, B; Ko, B R; Koay, S A; Kondo, K; Kong, D J; Konigsberg, J; Korytov, A; Kotwal, A V; Kraus, J; Kreps, M; Kroll, J; Krumnack, N; Kruse, M; Krutelyov, V; Kubo, T; Kuhlmann, S E; Kuhr, T; Kulkarni, N P; Kusakabe, Y; Kwang, S; Laasanen, A T; Labarga, L; Lai, S; Lami, S; Lammel, S; Lancaster, M; Lander, R L; Lannon, K; Lath, A; Latino, G; Lazzizzera, I; LeCompte, T; Lee, J; Lee, J; Lee, Y J; Lee, S W; Lefèvre, R; Leonardo, N; Leone, S; Levy, S; Lewis, J D; Lin, C; Lin, C S; Linacre, J; Lindgren, M; Lipeles, E; Lister, A; Litvintsev, D O; Liu, C; Liu, T; Lockyer, N S; Loginov, A; Loreti, M; Lovas, L; Lu, R-S; Lucchesi, D; Lueck, J; Luci, C; Lujan, P; Lukens, P; Lungu, G; Lyons, L; Lys, J; Lysak, R; Lytken, E; Mack, P; MacQueen, D; Madrak, R; Maeshima, K; Makhoul, K; Maki, T; Maksimovic, P; Malde, S; Malik, S; Manca, G; Manousakis, A; Margaroli, F; Marino, C; Marino, C P; Martin, A; Martin, M; Martin, V; Martínez, M; Martínez-Ballarín, R; Maruyama, T; Mastrandrea, P; Masubuchi, T; Mattson, M E; Mazzanti, P; McFarland, K S; McIntyre, P; McNulty, R; Mehta, A; Mehtala, P; Menzemer, S; Menzione, A; Merkel, P; Mesropian, C; Messina, A; Miao, T; Miladinovic, N; Miles, J; Miller, R; Mills, C; Milnik, M; Mitra, A; Mitselmakher, G; Miyake, H; Moed, S; Moggi, N; Moon, C S; Moore, R; Morello, M; Movilla Fernandez, P; Mülmenstädt, J; Mukherjee, A; Muller, Th; Mumford, R; Murat, P; Mussini, M; Nachtman, J; Nagai, Y; Nagano, A; Naganoma, J; Nakamura, K; Nakano, I; Napier, A; Necula, V; Neu, C; Neubauer, M S; Nielsen, J; Nodulman, L; Norman, M; Norniella, O; Nurse, E; Oh, S H; Oh, Y D; Oksuzian, I; Okusawa, T; Oldeman, R; Orava, R; Osterberg, K; Pagan Griso, S; Pagliarone, C; Palencia, E; Papadimitriou, V; Papaikonomou, A; Paramonov, A A; Parks, B; Pashapour, S; Patrick, J; Pauletta, G; Paulini, M; Paus, C; Pellett, D E; Penzo, A; Phillips, T J; Piacentino, G; Piedra, J; Pinera, L; Pitts, K; Plager, C; Pondrom, L; Portell, X; Poukhov, O; Pounder, N; Prakoshyn, F; Pronko, A; Proudfoot, J; Ptohos, F; Punzi, G; Pursley, J; Rademacker, J; Rahaman, A; Ramakrishnan, V; Ranjan, N; Redondo, I; Reisert, B; Rekovic, V; Renton, P; Rescigno, M; Richter, S; Rimondi, F; Ristori, L; Robson, A; Rodrigo, T; Rogers, E; Rolli, S; Roser, R; Rossi, M; Rossin, R; Roy, P; Ruiz, A; Russ, J; Rusu, V; Saarikko, H; Safonov, A; Sakumoto, W K; Salamanna, G; Saltó, O; Santi, L; Sarkar, S; Sartori, L; Sato, K; Savoy-Navarro, A; Scheidle, T; Schlabach, P; Schmidt, E E; Schmidt, M A; Schmidt, M P; Schmitt, M; Schwarz, T; Scodellaro, L; Scott, A L; Scribano, A; Scuri, F; Sedov, A; Seidel, S; Seiya, Y; Semenov, A; Sexton-Kennedy, L; Sfyrla, A; Shalhout, S Z; Shapiro, M D; Shears, T; Shepard, P F; Sherman, D; Shimojima, M; Shochet, M; Shon, Y; Shreyber, I; Sidoti, A; Sinervo, P; Sisakyan, A; Slaughter, A J; Slaunwhite, J; Sliwa, K; Smith, J R; Snider, F D; Snihur, R; Soderberg, M; Soha, A; Somalwar, S; Sorin, V; Spalding, J; Spinella, F; Spreitzer, T; Squillacioti, P; Stanitzki, M; St Denis, R; Stelzer, B; Stelzer-Chilton, O; Stentz, D; Strologas, J; Stuart, D; Suh, J S; Sukhanov, A; Sun, H; Suslov, I; Suzuki, T; Taffard, A; Takashima, R; Takeuchi, Y; Tanaka, R; Tecchio, M; Teng, P K; Terashi, K; Thom, J; Thompson, A S; Thompson, G A; Thomson, E; Tipton, P; Tiwari, V; Tkaczyk, S; Toback, D; Tokar, S; Tollefson, K; Tomura, T; Tonelli, D; Torre, S; Torretta, D; Tourneur, S; Trischuk, W; Tu, Y; Turini, N; Ukegawa, F; Uozumi, S; Vallecorsa, S; van Remortel, N; Varganov, A; Vataga, E; Vázquez, F; Velev, G; Vellidis, C; Veszpremi, V; Vidal, M; Vidal, R; Vila, I; Vilar, R; Vine, T; Vogel, M; Volobouev, I; Volpi, G; Würthwein, F; Wagner, P; Wagner, R G; Wagner, R L; Wagner-Kuhr, J; Wagner, W; Wakisaka, T; Wallny, R; Wang, S M; Warburton, A; Waters, D; Weinberger, M; Wester, W C; Whitehouse, B; Whiteson, D; Wicklund, A B; Wicklund, E; Williams, G; Williams, H H; Wilson, P; Winer, B L; Wittich, P; Wolbers, S; Wolfe, C; Wright, T; Wu, X; Wynne, S M; Yagil, A; Yamamoto, K; Yamaoka, J; Yamashita, T; Yang, C; Yang, U K; Yang, Y C; Yao, W M; Yeh, G P; Yoh, J; Yorita, K; Yoshida, T; Yu, G B; Yu, I; Yu, S S; Yun, J C; Zanello, L; Zanetti, A; Zaw, I; Zhang, X; Zheng, Y; Zucchelli, S

    2008-04-25

    This Letter describes the first determination of bounds on the CP-violation parameter 2beta(s) using B(s)(0) decays in which the flavor of the bottom meson at production is identified. The result is based on approximately 2000 B(s)(0)-->J/psiphi decays reconstructed in a 1.35 fb(-1) data sample collected with the CDF II detector using pp collisions produced at the Fermilab Tevatron. We report confidence regions in the two-dimensional space of 2beta(s) and the decay-width difference DeltaGamma. Assuming the standard model predictions of 2beta(s) and DeltaGamma, the probability of a deviation as large as the level of the observed data is 15%, corresponding to 1.5 Gaussian standard deviations.

  14. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pasek, Marta; Boeggeman, Elizabeth; Ramakrishnan, Boopathy

    The expression of recombinant proteins in Escherichia coli often leads to inactive aggregated proteins known as the inclusion bodies. To date, the best available tool has been the use of fusion tags, including the carbohydrate-binding protein; e.g., the maltose-binding protein (MBP) that enhances the solubility of recombinant proteins. However, none of these fusion tags work universally with every partner protein. We hypothesized that galectins, which are also carbohydrate-binding proteins, may help as fusion partners in folding the mammalian proteins in E. coli. Here we show for the first time that a small soluble lectin, human galectin-1, one member of amore » large galectin family, can function as a fusion partner to produce soluble folded recombinant human glycosyltransferase, {beta}-1,4-galactosyltransferase-7 ({beta}4Gal-T7), in E. coli. The enzyme {beta}4Gal-T7 transfers galactose to xylose during the synthesis of the tetrasaccharide linker sequence attached to a Ser residue of proteoglycans. Without a fusion partner, {beta}4Gal-T7 is expressed in E. coli as inclusion bodies. We have designed a new vector construct, pLgals1, from pET-23a that includes the sequence for human galectin-1, followed by the Tev protease cleavage site, a 6x His-coding sequence, and a multi-cloning site where a cloned gene is inserted. After lactose affinity column purification of galectin-1-{beta}4Gal-T7 fusion protein, the unique protease cleavage site allows the protein {beta}4Gal-T7 to be cleaved from galectin-1 that binds and elutes from UDP-agarose column. The eluted protein is enzymatically active, and shows CD spectra comparable to the folded {beta}4Gal-T1. The engineered galectin-1 vector could prove to be a valuable tool for expressing other proteins in E. coli.« less

  15. Single molecule fluorescence imaging as a technique for barium tagging in neutrinoless double beta decay

    NASA Astrophysics Data System (ADS)

    Jones, B. J. P.; McDonald, A. D.; Nygren, D. R.

    2016-12-01

    Background rejection is key to success for future neutrinoless double beta decay experiments. To achieve sensitivity to effective Majorana lifetimes of ~ 1028 years, backgrounds must be controlled to better than 0.1 count per ton per year, beyond the reach of any present technology. In this paper we propose a new method to identify the birth of the barium daughter ion in the neutrinoless double beta decay of 136Xe. The method adapts Single Molecule Fluorescent Imaging, a technique from biochemistry research with demonstrated single ion sensitivity. We explore possible SMFI dyes suitable for the problem of barium ion detection in high pressure xenon gas, and develop a fiber-coupled sensing system with which we can detect the presence of bulk Ba++ ions remotely. We show that our sensor produces signal-to-background ratios as high as 85 in response to Ba++ ions when operated in aqueous solution. We then describe the next stage of this R&D program, which will be to demonstrate chelation and fluorescence in xenon gas. If a successful barium ion tag can be developed using SMFI adapted for high pressure xenon gas detectors, the first essentially zero background, ton-scale neutrinoless double beta decay technology could be realized.

  16. Cloning and expression of antibacterial goat lactoferricin from Escherichia coli AD494(DE3)pLysS expression system.

    PubMed

    Chen, Gen-Hung; Yin, Li-Jung; Chiang, I-Hua; Jiang, Shann-Tzong

    2008-12-01

    Goat lactoferricin (GLfcin), an antibacterial peptide, is released from the N terminus of goat lactoferrin by pepsin digestion. Two GLfcin-related cDNAs, GLfcin L and GLfcin S, encoding Ala20-Ser60 and Ser36-Ser60 of goat lactoferrin, respectively, were cloned into the pET-23a(+) expression vector upstream from (His)6-Tag gene and transformed into Escherichia coli AD494(DE3)pLysS expression host. After being induced by isopropyl-beta-D-thiogalactopyranoside (IPTG), two (His)6-Tag fused recombinant lactoferricins, GLfcin L-His*Tag and GLfcin S-His*Tag, were expressed in soluble form within the E. coli cytoplasm. The GLfcin L-His*Tag and GLfcin S-His*Tag were purified using HisTrap affinity chromatography. According to an antibacterial activity assay using the agar diffusion method, GLfcin L-His*Tag had antibacterial activity against E. coli BCRC 11549, Staphylococcus aureus BCRC 25923, and Propionibacterium acnes BCRC 10723, while GLfcin S-His*Tag was able to inhibit the growth of E. coli BCRC 11549 and P. acnes BCRC 10723. These two recombinant lactoferricins behaved as thermostable peptides, which could retain their activity for up to 30 min of exposure at 100 degrees C.

  17. Cloning, expression, and characterization of a peculiar choline-binding beta-galactosidase from Streptococcus mitis.

    PubMed

    Campuzano, Susana; Serra, Beatriz; Llull, Daniel; García, José L; García, Pedro

    2009-09-01

    A Streptococcus mitis genomic DNA fragment carrying the SMT1224 gene encoding a putative beta-galactosidase was identified, cloned, and expressed in Escherichia coli. This gene encodes a protein 2,411 amino acids long with a predicted molecular mass of 268 kDa. The deduced protein contains an N-terminal signal peptide and a C-terminal choline-binding domain consisting of five consensus repeats, which facilitates the anchoring of the secreted enzyme to the cell wall. The choline-binding capacity of the protein facilitates its purification using DEAE-cellulose affinity chromatography, although its complete purification was achieved by constructing a His-tagged fusion protein. The recombinant protein was characterized as a monomeric beta-galactosidase showing a specific activity of around 2,500 U/mg of protein, with optimum temperature and pH ranges of 30 to 40 degrees C and 6.0 to 6.5, respectively. Enzyme activity is not inhibited by glucose, even at 200 mM, and remains highly stable in solution or immobilized at room temperature in the absence of protein stabilizers. In S. mitis, the enzyme was located attached to the cell surface, but a significant activity was also detected in the culture medium. This novel enzyme represents the first beta-galactosidase having a modular structure with a choline-binding domain, a peculiar property that can also be useful for some biotechnological applications.

  18. A novel expression system for intracellular production and purification of recombinant affinity-tagged proteins in Aspergillus niger.

    PubMed

    Roth, Andreas H F J; Dersch, Petra

    2010-03-01

    A set of different integrative expression vectors for the intracellular production of recombinant proteins with or without affinity tag in Aspergillus niger was developed. Target genes can be expressed under the control of the highly efficient, constitutive pkiA promoter or the novel sucrose-inducible promoter of the beta-fructofuranosidase (sucA) gene of A. niger in the presence or absence of alternative carbon sources. All expression plasmids contain an identical multiple cloning sequence that allows parallel construction of N- or C-terminally His6- and StrepII-tagged versions of the target proteins. Production of two heterologous model proteins, the green fluorescence protein and the Thermobifida fusca hydrolase, proved the functionality of the vector system. Efficient production and easy detection of the target proteins as well as their fast purification by a one-step affinity chromatography, using the His6- or StrepII-tag sequence, was demonstrated.

  19. Demonstration of Single-Barium-Ion Sensitivity for Neutrinoless Double-Beta Decay Using Single-Molecule Fluorescence Imaging.

    PubMed

    McDonald, A D; Jones, B J P; Nygren, D R; Adams, C; Álvarez, V; Azevedo, C D R; Benlloch-Rodríguez, J M; Borges, F I G M; Botas, A; Cárcel, S; Carrión, J V; Cebrián, S; Conde, C A N; Díaz, J; Diesburg, M; Escada, J; Esteve, R; Felkai, R; Fernandes, L M P; Ferrario, P; Ferreira, A L; Freitas, E D C; Goldschmidt, A; Gómez-Cadenas, J J; González-Díaz, D; Gutiérrez, R M; Guenette, R; Hafidi, K; Hauptman, J; Henriques, C A O; Hernandez, A I; Hernando Morata, J A; Herrero, V; Johnston, S; Labarga, L; Laing, A; Lebrun, P; Liubarsky, I; López-March, N; Losada, M; Martín-Albo, J; Martínez-Lema, G; Martínez, A; Monrabal, F; Monteiro, C M B; Mora, F J; Moutinho, L M; Muñoz Vidal, J; Musti, M; Nebot-Guinot, M; Novella, P; Palmeiro, B; Para, A; Pérez, J; Querol, M; Repond, J; Renner, J; Riordan, S; Ripoll, L; Rodríguez, J; Rogers, L; Santos, F P; Dos Santos, J M F; Simón, A; Sofka, C; Sorel, M; Stiegler, T; Toledo, J F; Torrent, J; Tsamalaidze, Z; Veloso, J F C A; Webb, R; White, J T; Yahlali, N

    2018-03-30

    A new method to tag the barium daughter in the double-beta decay of ^{136}Xe is reported. Using the technique of single molecule fluorescent imaging (SMFI), individual barium dication (Ba^{++}) resolution at a transparent scanning surface is demonstrated. A single-step photobleach confirms the single ion interpretation. Individual ions are localized with superresolution (∼2  nm), and detected with a statistical significance of 12.9σ over backgrounds. This lays the foundation for a new and potentially background-free neutrinoless double-beta decay technology, based on SMFI coupled to high pressure xenon gas time projection chambers.

  20. Demonstration of Single-Barium-Ion Sensitivity for Neutrinoless Double-Beta Decay Using Single-Molecule Fluorescence Imaging

    NASA Astrophysics Data System (ADS)

    McDonald, A. D.; Jones, B. J. P.; Nygren, D. R.; Adams, C.; Álvarez, V.; Azevedo, C. D. R.; Benlloch-Rodríguez, J. M.; Borges, F. I. G. M.; Botas, A.; Cárcel, S.; Carrión, J. V.; Cebrián, S.; Conde, C. A. N.; Díaz, J.; Diesburg, M.; Escada, J.; Esteve, R.; Felkai, R.; Fernandes, L. M. P.; Ferrario, P.; Ferreira, A. L.; Freitas, E. D. C.; Goldschmidt, A.; Gómez-Cadenas, J. J.; González-Díaz, D.; Gutiérrez, R. M.; Guenette, R.; Hafidi, K.; Hauptman, J.; Henriques, C. A. O.; Hernandez, A. I.; Hernando Morata, J. A.; Herrero, V.; Johnston, S.; Labarga, L.; Laing, A.; Lebrun, P.; Liubarsky, I.; López-March, N.; Losada, M.; Martín-Albo, J.; Martínez-Lema, G.; Martínez, A.; Monrabal, F.; Monteiro, C. M. B.; Mora, F. J.; Moutinho, L. M.; Muñoz Vidal, J.; Musti, M.; Nebot-Guinot, M.; Novella, P.; Palmeiro, B.; Para, A.; Pérez, J.; Querol, M.; Repond, J.; Renner, J.; Riordan, S.; Ripoll, L.; Rodríguez, J.; Rogers, L.; Santos, F. P.; dos Santos, J. M. F.; Simón, A.; Sofka, C.; Sorel, M.; Stiegler, T.; Toledo, J. F.; Torrent, J.; Tsamalaidze, Z.; Veloso, J. F. C. A.; Webb, R.; White, J. T.; Yahlali, N.; NEXT Collaboration

    2018-03-01

    A new method to tag the barium daughter in the double-beta decay of Xe 136 is reported. Using the technique of single molecule fluorescent imaging (SMFI), individual barium dication (Ba++ ) resolution at a transparent scanning surface is demonstrated. A single-step photobleach confirms the single ion interpretation. Individual ions are localized with superresolution (˜2 nm ), and detected with a statistical significance of 12.9 σ over backgrounds. This lays the foundation for a new and potentially background-free neutrinoless double-beta decay technology, based on SMFI coupled to high pressure xenon gas time projection chambers.

  1. Reliability of an ordinal rating system for assessing the amount of mud and feces (tag) on cattle hides at slaughter.

    PubMed

    Jordan, D; McEwen, S A; Wilson, J B; McNab, W B; Lammerding, A M

    1999-05-01

    A study was conducted to provide a quantitative description of the amount of tag (mud, soil, and bedding) adhered to the hides of feedlot beef cattle and to appraise the statistical reliability of a subjective rating system for assessing this trait. Initially, a single rater obtained baseline data by assessing 2,417 cattle for 1 month at an Ontario beef processing plant. Analysis revealed that there was a strong tendency for animals within sale-lots to have a similar total tag score (intralot correlation = 0.42). Baseline data were summarized by fitting a linear model describing an individual's total tag score as the sum of their lot mean tag score (LMTS) plus an amount representing normal variation within the lot. LMTSs predicted by the linear model were adequately described by a beta distribution with parameters nu = 3.12 and omega = 5.82 scaled to fit on the 0-to-9 interval. Five raters, trained in use of the tag scoring system, made 1,334 tag score observations in a commercial abattoir, allowing reliability to be assessed at the individual level and at the lot level. High values for reliability were obtained for individual total tag score (0.84) and lot total tag score (0.83); these values suggest that the tag scoring system could be used in the marketing and slaughter of Ontario beef cattle to improve the cleanliness of animals presented for slaughter in an effort to control the entry of microbial contamination into abattoirs. Implications for the use of the tag scoring system in research are discussed.

  2. Current test results for the Athena radar responsive tag

    NASA Astrophysics Data System (ADS)

    Ormesher, Richard C.; Martinez, Ana; Plummer, Kenneth W.; Erlandson, David; Delaware, Sheri; Clark, David R.

    2006-05-01

    Sandia National Laboratories has teamed with General Atomics and Sierra Monolithics to develop the Athena tag for the Army's Radar Tag Engagement (RaTE) program. The radar-responsive Athena tag can be used for Blue Force tracking and Combat Identification (CID) as well as data collection, identification, and geolocation applications. The Athena tag is small (~4.5" x 2.4" x 4.2"), battery-powered, and has an integral antenna. Once remotely activated by a Synthetic Aperture Radar (SAR) or Moving Target Indicator (MTI) radar, the tag transponds modulated pulses to the radar at a low transmit power. The Athena tag can operate Ku-band and X-band airborne SAR and MTI radars. This paper presents results from current tag development testing activities. Topics covered include recent field tests results from the AN/APY-8 Lynx, F16/APG-66, and F15E/APG-63 V(1) radars and other Fire Control radars. Results show that the Athena tag successfully works with multiple radar platforms, in multiple radar modes, and for multiple applications. Radar-responsive tags such as Athena have numerous applications in military and government arenas. Military applications include battlefield situational awareness, combat identification, targeting, personnel recovery, and unattended ground sensors. Government applications exist in nonproliferation, counter-drug, search-and-rescue, and land-mapping activities.

  3. Multiple Object Based RFID System Using Security Level

    NASA Astrophysics Data System (ADS)

    Kim, Jiyeon; Jung, Jongjin; Ryu, Ukjae; Ko, Hoon; Joe, Susan; Lee, Yongjun; Kim, Boyeon; Chang, Yunseok; Lee, Kyoonha

    2007-12-01

    RFID systems are increasingly applied for operational convenience in wide range of industries and individual life. However, it is uneasy for a person to control many tags because common RFID systems have the restriction that a tag used to identify just a single object. In addition, RFID systems can make some serious problems in violation of privacy and security because of their radio frequency communication. In this paper, we propose a multiple object RFID tag which can keep multiple object identifiers for different applications in a same tag. The proposed tag allows simultaneous access for their pair applications. We also propose an authentication protocol for multiple object tag to prevent serious problems of security and privacy in RFID applications. Especially, we focus on efficiency of the authentication protocol by considering security levels of applications. In the proposed protocol, the applications go through different authentication procedures according to security level of the object identifier stored in the tag. We implemented the proposed RFID scheme and made experimental results about efficiency and stability for the scheme.

  4. Expression of beta-expansins is correlated with internodal elongation in deepwater rice.

    PubMed

    Lee, Y; Kende, H

    2001-10-01

    Fourteen putative rice (Oryza sativa) beta-expansin genes, Os-EXPB1 through Os-EXPB14, were identified in the expressed sequence tag and genomic databases. The DNA and deduced amino acid sequences are highly conserved in all 14 beta-expansins. They have a series of conserved C (cysteine) residues in the N-terminal half of the protein, an HFD (histidine-phenylalanine-aspartate) motif in the central region, and a series of W (tryptophan) residues near the carboxyl terminus. Five beta-expansin genes are expressed in deepwater rice internodes, with especially high transcript levels in the growing region. Expression of four beta-expansin genes in the internode was induced by treatment with gibberellin and by wounding. The wound response resulted from excising stem sections or from piercing pinholes into the stem of intact plants. The level of wound-induced beta-expansin transcripts declined rapidly 5 h after cutting of stem sections. We conclude that the expression of beta-expansin genes is correlated with rapid elongation of deepwater rice internodes, it is induced by gibberellin and wounding, and wound-induced beta-expansin mRNA appears to turn over rapidly.

  5. A study of time-dependent CP-violating asymmetries in B0->J/psiK0S and B0->psi(2S)K0S decays

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Barrera, Barbara

    We present a preliminary measurement of time-dependent CP-violating asymmetries in B{sup 0} {yields} J/{psi} K{sub S}{sup 0} and B{sup 0} {yields} {psi}(2S)K{sub S}{sup 0} decays recorded by the BABAR detector at the PEP-II asymmetric-energy B Factory at SLAC. The data sample consists of 9.0 fb{sup -1} collected at the {Upsilon}(4S) resonance and 0.8 fb{sup -1} off-resonance. One of the neutral B mesons, produced in pairs at the {Upsilon}(4S), is fully reconstructed. The flavor of the other neutral B meson is tagged at the time of its decay, mainly with the charge of identified leptons and kaons. A neural network taggingmore » algorithm is used to recover events without a clear lepton or kaon tag. The time difference between the decays is determined by measuring the distance between the decay vertices. Wrong-tag probabilities and the time resolution function are measured with samples of fully-reconstructed semileptonic and hadronic neutral B final states. The value of the asymmetry amplitude, sin2{beta}, is determined from a maximum likelihood fit to the time distribution of 120 tagged B{sup 0} {yields} J/{psi} K{sub S}{sup 0} and B{sup 0} {yields} {psi}(2S) K{sub S}{sup 0} candidates to be sin2{beta} = 0.12 {+-} 0.37(stat) {+-} 0.09(syst) (preliminary).« less

  6. Metabolic engineering of Saccharomyces cerevisiae for overproduction of triacylglycerols.

    PubMed

    Ferreira, Raphael; Teixeira, Paulo Gonçalves; Gossing, Michael; David, Florian; Siewers, Verena; Nielsen, Jens

    2018-06-01

    Triacylglycerols (TAGs) are valuable versatile compounds that can be used as metabolites for nutrition and health, as well as feedstocks for biofuel production. Although Saccharomyces cerevisiae is the favored microbial cell factory for industrial production of biochemicals, it does not produce large amounts of lipids and TAGs comprise only ~1% of its cell dry weight. Here, we engineered S. cerevisiae to reorient its metabolism for overproduction of TAGs, by regulating lipid droplet associated-proteins involved in TAG synthesis and hydrolysis. We implemented a push-and-pull strategy by overexpressing genes encoding a deregulated acetyl-CoA carboxylase, ACC1 S659A/S1157A (ACC1**) , as well as the last two steps of TAG formation: phosphatidic phosphatase ( PAH1 ) and diacylglycerol acyltransferase ( DGA1 ), ultimately leading to 129 mg∙gCDW -1 of TAGs. Disruption of TAG lipase genes TGL3 , TGL4 , TGL5 and sterol acyltransferase gene ARE1 increased the TAG content to 218 mg∙gCDW -1 . Further disruption of the beta-oxidation by deletion of POX1 , as well as glycerol-3-phosphate utilization through deletion of GUT2 , did not affect TAGs levels. Finally, disruption of the peroxisomal fatty acyl-CoA transporter PXA1 led to accumulation of 254 mg∙gCDW -1 . The TAG levels achieved here are the highest titer reported in S. cerevisiae , reaching 27.4% of the maximum theoretical yield in minimal medium with 2% glucose. This work shows the potential of using an industrially established and robust yeast species for high level lipid production.

  7. Demonstration of Single-Barium-Ion Sensitivity for Neutrinoless Double-Beta Decay Using Single-Molecule Fluorescence Imaging

    DOE PAGES

    McDonald, A. D.; Jones, B. J. P.; Nygren, D. R.; ...

    2018-03-26

    A new method to tag the barium daughter in the double beta decay ofmore » $$^{136}$$Xe is reported. Using the technique of single molecule fluorescent imaging (SMFI), individual barium dication (Ba$$^{++}$$) resolution at a transparent scanning surface has been demonstrated. A single-step photo-bleach confirms the single ion interpretation. Individual ions are localized with super-resolution ($$\\sim$$2~nm), and detected with a statistical significance of 12.9~$$\\sigma$$ over backgrounds. This lays the foundation for a new and potentially background-free neutrinoless double beta decay technology, based on SMFI coupled to high pressure xenon gas time projection chambers.« less

  8. HaloTag Technology: A Versatile Platform for Biomedical Applications

    PubMed Central

    2015-01-01

    Exploration of protein function and interaction is critical for discovering links among genomics, proteomics, and disease state; yet, the immense complexity of proteomics found in biological systems currently limits our investigational capacity. Although affinity and autofluorescent tags are widely employed for protein analysis, these methods have been met with limited success because they lack specificity and require multiple fusion tags and genetic constructs. As an alternative approach, the innovative HaloTag protein fusion platform allows protein function and interaction to be comprehensively analyzed using a single genetic construct with multiple capabilities. This is accomplished using a simplified process, in which a variable HaloTag ligand binds rapidly to the HaloTag protein (usually linked to the protein of interest) with high affinity and specificity. In this review, we examine all current applications of the HaloTag technology platform for biomedical applications, such as the study of protein isolation and purification, protein function, protein–protein and protein–DNA interactions, biological assays, in vitro cellular imaging, and in vivo molecular imaging. In addition, novel uses of the HaloTag platform are briefly discussed along with potential future applications. PMID:25974629

  9. Serum retinol, alpha-tocopherol, and beta-carotene levels are not altered by excess ingestion of diacylglycerol-containing plant sterol esters.

    PubMed

    Saito, Shinichiro; Tomonobu, Kazuichi; Kudo, Naoto; Shiiba, Daisuke; Hase, Tadashi; Tokimitsu, Ichiro

    2006-01-01

    Diacylglycerol (DAG) suppresses the postprandial increase in serum triglycerides, and has antiobesity effects. On the other hand, plant sterol esters (PSE) lower serum cholesterol levels in hypercholesterolemia. Thus, DAG-containing PSE (PSE/DAG) would be expected to maintain an appropriate serum cholesterol level and decrease the risk of arteriosclerotic disorders. Several recent studies, however, report negative effects of PSE on serum fat-soluble (pro)vitamin levels. The objective of this study was to investigate the effect of PSE/DAG on serum retinol, beta-carotene, and alpha-tocopherol levels using a threefold excess of the effective dose obtained in our previous study. A randomized placebo-controlled double-blind parallel study was performed in healthy and mildly hypercholesterolemic subjects, in which the subjects ingested 1.2 g PSE/30 g DAG for 2 weeks in the form of mayonnaise-type products. Triacylglycerol (TAG) mayonnaise was used as a control. There were no subjective adverse effects or changes in serum retinol, alpha-tocopherol, and beta-carotene levels, abdominal symptoms, hematologic values, or blood biochemical values. Ingestion of a threefold excess of PSE/DAG for 2 weeks had no adverse effects compared to ingestion of conventional TAG mayonnaise. Copyright 2006 S. Karger AG, Basel.

  10. Direct detection of ligand binding to Sepharose-immobilised protein using saturation transfer double difference (STDD) NMR spectroscopy

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Haselhorst, Thomas; Muenster-Kuehnel, Anja K.; Oschlies, Melanie

    2007-08-10

    We report an easy and direct application of 'Saturation Transfer Double Difference' (STDD) NMR spectroscopy to identify ligands that bind to a Sepharose-immobilised target protein. The model protein, cytidine 5'-monophosphate sialic acid (CMP-Sia) synthetase, was expressed as a Strep-Tag II fusion protein and immobilised on Strep-Tactin Sepharose. STD NMR experiments of the protein-enriched Sepharose matrix in the presence of a binding ligand (cytidine 5'-triphosphate, CTP) and a non-binding ligand ({alpha}/{beta}-glucose) clearly show that CTP binds to the immobilised enzyme, whereas glucose has no affinity. This approach has three major advantages: (a) only low quantities of protein are required, (b) nomore » specialised NMR technology or the application of additional data analysis by non-routine methods is required, and (c) easy multiple use of the immobilised protein is available.« less

  11. R(-)-4-(3-Isothiocyanatopyrrolidin-1-yl)-7-(N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole, a fluorescent chiral tagging reagent: sensitive resolution of chiral amines and amino acids by reversed-phase liquid chromatography.

    PubMed

    Toyo'oka, T; Jin, D; Tomoi, N; Oe, T; Hiranuma, H

    2001-02-01

    The usefulness of R(-)-4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole [R(-)-DBD-PyNCS], a fluorescent chiral tagging reagent, for the determination of racemic amines and amino acids, was studied. The reagent reacted with beta-blockers selected as representative secondary amines to produce corresponding fluorescent diastereomers (excitation at 460 nm and emission at 550 nm). The yields of the derivatization reaction were dependent on the stereostructure arround the NH group in beta-blockers. The resulting diastereomers were completely separated with single chromatographic run using linear gradient elutions by reversed-phase chromatography. R(-)-DBD-PyNCS was also applied to the determination of DL-amino acid, considered to be one of the primary amines, in human urine and foodstuffs. DL-amino acids tested equally reacted with the reagent, and the thiocarbamoyl derivatives were separated with an ODS column. The epimerization during the derivatization reaction was negligible judging from the resolution of opposite diastereomers on the chromatogram. The occurence of D-amino acids (D-Ala, D-Ser, D-Asp and/or D-Glu) was identified in the samples tested. The structures and the purities were elucidated with on-line HPLC-MS. The chiral reagent possessing an isothiocyanate group (-NCS) in the structure seems to be applicable to continuous sequential analysis of peptides containing D-amino acids. The thiocarbamoyl derivatives obtained from the reaction with DL-amino acids were converted to thiohydantoins via thiazolinones in acidic medium. The thiohydantoins produced from acidic, basic, neutral, hydroxyl and aromatic amino acids were completely separated with isocratic elutions using acidic mobile phase containing 0.1% TFA. The separations were sufficient for the identification of DL-amino acid in peptide sequences. Although the epimerization during the conversion reaction to thiohydantoins was not avoidable, the descrimination of D- and L-configuration was demonstrated with some commercially available peptides such as beta-lipotropin and [D-Ala2]-deltorphin II. The Edman degaradation method using R(-)-DBD-PyNCS was also adopted to autoanlaysis by gas-phase sequencer. The separation and the detection (UV 254 nm) conditions of the derivatives were used without any change from those for the Edman degradation method using PITC as the tagging reagent. The three DL-amino acid residues (Tyr, Ala and Gly) in [L-Ala2]-leucine-enkephalin and [D-Ala2]-leucine-enkephalin were perfectly identidied with the autoanalysis.

  12. High-throughput purification of recombinant proteins using self-cleaving intein tags.

    PubMed

    Coolbaugh, M J; Shakalli Tang, M J; Wood, D W

    2017-01-01

    High throughput methods for recombinant protein production using E. coli typically involve the use of affinity tags for simple purification of the protein of interest. One drawback of these techniques is the occasional need for tag removal before study, which can be hard to predict. In this work, we demonstrate two high throughput purification methods for untagged protein targets based on simple and cost-effective self-cleaving intein tags. Two model proteins, E. coli beta-galactosidase (βGal) and superfolder green fluorescent protein (sfGFP), were purified using self-cleaving versions of the conventional chitin-binding domain (CBD) affinity tag and the nonchromatographic elastin-like-polypeptide (ELP) precipitation tag in a 96-well filter plate format. Initial tests with shake flask cultures confirmed that the intein purification scheme could be scaled down, with >90% pure product generated in a single step using both methods. The scheme was then validated in a high throughput expression platform using 24-well plate cultures followed by purification in 96-well plates. For both tags and with both target proteins, the purified product was consistently obtained in a single-step, with low well-to-well and plate-to-plate variability. This simple method thus allows the reproducible production of highly pure untagged recombinant proteins in a convenient microtiter plate format. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Chemical rescue of cleft palate and midline defects in conditional GSK-3beta mice.

    PubMed

    Liu, Karen J; Arron, Joseph R; Stankunas, Kryn; Crabtree, Gerald R; Longaker, Michael T

    2007-03-01

    Glycogen synthase kinase-3beta (GSK-3beta) has integral roles in a variety of biological processes, including development, diabetes, and the progression of Alzheimer's disease. As such, a thorough understanding of GSK-3beta function will have a broad impact on human biology and therapeutics. Because GSK-3beta interacts with many different pathways, its specific developmental roles remain unclear. We have discovered a genetic requirement for GSK-3beta in midline development. Homozygous null mice display cleft palate, incomplete fusion of the ribs at the midline and bifid sternum as well as delayed sternal ossification. Using a chemically regulated allele of GSK-3beta (ref. 6), we have defined requirements for GSK-3beta activity during discrete temporal windows in palatogenesis and skeletogenesis. The rapamycin-dependent allele of GSK-3beta produces GSK-3beta fused to a tag, FRB* (FKBP/rapamycin binding), resulting in a rapidly destabilized chimaeric protein. In the absence of drug, GSK-3beta(FRB)*(/FRB)* mutants appear phenotypically identical to GSK-3beta-/- mutants. In the presence of drug, GSK-3betaFRB* is rapidly stabilized, restoring protein levels and activity. Using this system, mutant phenotypes were rescued by restoring endogenous GSK-3beta activity during two distinct periods in gestation. This technology provides a powerful tool for defining windows of protein function during development.

  14. Immunoassay for Visualization of Protein-Protein Interactions on Ni-Nitrilotriacetate Support: Example of a Laboratory Exercise with Recombinant Heterotrimeric G[alpha][subscript i2][beta][subscript 1[gamma]2] Tagged by Hexahistidine from sf9 Cells

    ERIC Educational Resources Information Center

    Bavec, Aljosa

    2004-01-01

    We have developed an "in vitro assay" for following the interaction between the [alpha][subscript i2] subunit and [beta][subscript 1[gamma]2] dimer from sf9 cells. This method is suitable for education purposes because it is easy, reliable, nonexpensive, can be applied for a big class of 20 students, and avoid the commonly used kinetic approach,…

  15. Analysis of association of clinical aspects and IL1B tagSNPs with severe preeclampsia.

    PubMed

    Leme Galvão, Larissa Paes; Menezes, Filipe Emanuel; Mendonca, Caio; Barreto, Ikaro; Alvim-Pereira, Claudia; Alvim-Pereira, Fabiano; Gurgel, Ricardo

    2016-01-01

    This study investigates the association between IL1B genotypes using a tag SNP (single polymorphism) approach, maternal and environmental factors in Brazilian women with severe preeclampsia. A case-control study with a total of 456 patients (169 preeclamptic women and 287 controls) was conducted in the two reference maternity hospitals of Sergipe state, Northeast Brazil. A questionnaire was administered and DNA was extracted to genotype the population for four tag SNPs of the IL1Beta: rs 1143643, rs 1143633, rs 1143634 and rs 1143630. Haplotype association analysis and p-values were calculated using the THESIAS test. Odds ratio (OR) estimation, confidence interval (CI) and multivariate logistic regression were performed. High pregestational body mass index (pre-BMI), first gestation, cesarean section, more than six medical visits, low level of consciousness on admission and TC and TT genotype in rs1143630 of IL1Beta showed association with the preeclamptic group in univariate analysis. After multivariate logistic regression pre-BMI, first gestation and low level of consciousness on admission remained associated. We identified an association between clinical variables and preeclampsia. Univariate analysis suggested that inflammatory process-related genes, such as IL1B, may be involved and should be targeted in further studies. The identification of the genetic background involved in preeclampsia host response modulation is mandatory in order to understand the preeclampsia process.

  16. Security techniques for prevention of rank manipulation in social tagging services including robotic domains.

    PubMed

    Choi, Okkyung; Jung, Hanyoung; Moon, Seungbin

    2014-01-01

    With smartphone distribution becoming common and robotic applications on the rise, social tagging services for various applications including robotic domains have advanced significantly. Though social tagging plays an important role when users are finding the exact information through web search, reliability and semantic relation between web contents and tags are not considered. Spams are making ill use of this aspect and put irrelevant tags deliberately on contents and induce users to advertise contents when they click items of search results. Therefore, this study proposes a detection method for tag-ranking manipulation to solve the problem of the existing methods which cannot guarantee the reliability of tagging. Similarity is measured for ranking the grade of registered tag on the contents, and weighted values of each tag are measured by means of synonym relevance, frequency, and semantic distances between tags. Lastly, experimental evaluation results are provided and its efficiency and accuracy are verified through them.

  17. DESIGN, SYNTHESIS, AND APPLICATION OF THE TRIMETHOPRIM-BASED CHEMICAL TAG FOR LIVE CELL IMAGING

    PubMed Central

    Jing, Chaoran; Cornish, Virginia W.

    2013-01-01

    Over the past decade chemical tags have been developed to complement the use of fluorescent proteins in live cell imaging. Chemical tags retain the specificity of protein labeling achieved with fluorescent proteins through genetic encoding, but provide smaller, more robust tags and modular use of organic fluorophores with high photon-output and tailored functionalities. The trimethoprim-based chemical tag (TMP-tag) was initially developed based on the high affinity interaction between E.coli dihydrofolatereductase and the antibiotic trimethoprim and subsequently rendered covalent and fluorogenic via proximity-induced protein labeling reactions. To date, the TMP-tag is one of the few chemical tags that enable intracellular protein labeling and high-resolution live cell imaging. Here we describe the general design, chemical synthesis, and application of TMP-tag for live cell imaging. Alternative protocols for synthesizing and using the covalent and the fluorogenic TMP-tags are also included. PMID:23839994

  18. Mocr: A novel fusion tag for enhancing solubility that is compatible with structural biology applications

    PubMed Central

    DelProposto, James; Majmudar, Chinmay Y.; Smith, Janet L.; Brown, William Clay

    2010-01-01

    A persistent problem in heterologous protein production is insolubility of the target protein when expressed to high level in the host cell. A widely employed strategy for overcoming this problem is the use of fusion tags. The best fusion tags promote solubility, may function as purification handles and either do not interfere with downstream applications or may be removed from the passenger protein preparation. A novel fusion tag is identified that meets these criteria. This fusion tag is a monomeric mutant of the Ocr protein (0.3 gene product) of bacteriophage T7. This fusion tag displays solubilizing activity with a variety of different passenger proteins. We show that it may be used as a purification handle similar to other fusion tags. Its small size and compact structure are compatible with its use in downstream applications of the passenger protein or it may be removed and purified away from the passenger protein. The use of monomeric Ocr (Mocr) as a complement to other fusion tags such as maltose-binding protein will provide greater flexibility in protein production and processing for a wide variety of protein applications. PMID:18824232

  19. Mocr: a novel fusion tag for enhancing solubility that is compatible with structural biology applications.

    PubMed

    DelProposto, James; Majmudar, Chinmay Y; Smith, Janet L; Brown, William Clay

    2009-01-01

    A persistent problem in heterologous protein production is insolubility of the target protein when expressed to high level in the host cell. A widely employed strategy for overcoming this problem is the use of fusion tags. The best fusion tags promote solubility, may function as purification handles and either do not interfere with downstream applications or may be removed from the passenger protein preparation. A novel fusion tag is identified that meets these criteria. This fusion tag is a monomeric mutant of the Ocr protein (0.3 gene product) of bacteriophage T7. This fusion tag displays solubilizing activity with a variety of different passenger proteins. We show that it may be used as a purification handle similar to other fusion tags. Its small size and compact structure are compatible with its use in downstream applications of the passenger protein or it may be removed and purified away from the passenger protein. The use of monomeric Ocr (Mocr) as a complement to other fusion tags such as maltose-binding protein will provide greater flexibility in protein production and processing for a wide variety of protein applications.

  20. Muon capture on light isotopes in Double Chooz

    NASA Astrophysics Data System (ADS)

    Strait, M.; Double Chooz Collaboration

    2017-09-01

    Using the Double Chooz reactor neutrino detector, we have measured the products of µ - capture on 12C, 13C, 14N and 16O. Over a period of 490 days, we collected 2.3 × 106 stopping cosmic µ -, of which 1.8 × 105 captured on these nuclei in the inner detector. The resulting isotopes were tagged using prompt neutron emission (when applicable), the subsequent beta decays, and, in some cases, β-delayed neutrons. Production of these βn isotopes, primarily 9Li, which are {{{ν _e}} \\over {{ν _μ }}} backgrounds, was found at a significance of 5.5σ. The probability of 9Li production per capture on natC is (2.4 ± 0.9(stat) ± 0.1(syst)) × 10-4. We have made the most precise measurement of the rate of 12C(µ -, ν)12B to date, 6.57 - 0.21 + 0.11 × {10^3}{{ }}{{{s}} - 1},{{ or }}≤ft( {17.35 - 0.59 + 0.35} \\right)% of nuclear captures. By tagging excited states emitting gammas, the ground state transition rate to 12B is found to be 5.68 - 0.23 + 0.14 × {10^3}{{ }}{{{s}} - 1}.

  1. Open semantic annotation of scientific publications using DOMEO.

    PubMed

    Ciccarese, Paolo; Ocana, Marco; Clark, Tim

    2012-04-24

    Our group has developed a useful shared software framework for performing, versioning, sharing and viewing Web annotations of a number of kinds, using an open representation model. The Domeo Annotation Tool was developed in tandem with this open model, the Annotation Ontology (AO). Development of both the Annotation Framework and the open model was driven by requirements of several different types of alpha users, including bench scientists and biomedical curators from university research labs, online scientific communities, publishing and pharmaceutical companies.Several use cases were incrementally implemented by the toolkit. These use cases in biomedical communications include personal note-taking, group document annotation, semantic tagging, claim-evidence-context extraction, reagent tagging, and curation of textmining results from entity extraction algorithms. We report on the Domeo user interface here. Domeo has been deployed in beta release as part of the NIH Neuroscience Information Framework (NIF, http://www.neuinfo.org) and is scheduled for production deployment in the NIF's next full release.Future papers will describe other aspects of this work in detail, including Annotation Framework Services and components for integrating with external textmining services, such as the NCBO Annotator web service, and with other textmining applications using the Apache UIMA framework.

  2. Open semantic annotation of scientific publications using DOMEO

    PubMed Central

    2012-01-01

    Background Our group has developed a useful shared software framework for performing, versioning, sharing and viewing Web annotations of a number of kinds, using an open representation model. Methods The Domeo Annotation Tool was developed in tandem with this open model, the Annotation Ontology (AO). Development of both the Annotation Framework and the open model was driven by requirements of several different types of alpha users, including bench scientists and biomedical curators from university research labs, online scientific communities, publishing and pharmaceutical companies. Several use cases were incrementally implemented by the toolkit. These use cases in biomedical communications include personal note-taking, group document annotation, semantic tagging, claim-evidence-context extraction, reagent tagging, and curation of textmining results from entity extraction algorithms. Results We report on the Domeo user interface here. Domeo has been deployed in beta release as part of the NIH Neuroscience Information Framework (NIF, http://www.neuinfo.org) and is scheduled for production deployment in the NIF’s next full release. Future papers will describe other aspects of this work in detail, including Annotation Framework Services and components for integrating with external textmining services, such as the NCBO Annotator web service, and with other textmining applications using the Apache UIMA framework. PMID:22541592

  3. Analysis of Tobyhanna Army Depot’s Radio Frequency Identification (RFID) Pilot Program: RFID as an Asset Management Tool

    DTIC Science & Technology

    2005-06-01

    each has advantages and disadvantages. 1. Active Tags The main difference between the two types of RFID tags is that active tags have an internal...battery source and passive tags do not. The advantage of the active tag is that it transmits its own signal, which increases the read range of the...applications provide a stand-alone solution that employs a specific usage application of RFID and has clear economic advantages over supply chain

  4. Technique for the application of a streamer-type fish tag

    USGS Publications Warehouse

    Joeris, Leonard S.

    1953-01-01

    Principal features of the technique are: attachment of the plastic tag by means of a nylon-thread loop prepared in advance of field work; use of a curved surgical needle with cutting edge and a split eye for application of the tag. The procedures for splitting the needle's eye and for applying the tag are described and illustrated by a series of photographs.

  5. Efficient recombinant expression and secretion of a thermostable GH26 mannan endo-1,4-beta-mannosidase from Bacillus licheniformis in Escherichia coli.

    PubMed

    Songsiriritthigul, Chomphunuch; Buranabanyat, Bancha; Haltrich, Dietmar; Yamabhai, Montarop

    2010-04-11

    Mannans are one of the key polymers in hemicellulose, a major component of lignocellulose. The Mannan endo-1,4-beta-mannosidase or 1,4-beta-D-mannanase (EC 3.2.1.78), commonly named beta-mannanase, is an enzyme that can catalyze random hydrolysis of beta-1,4-mannosidic linkages in the main chain of mannans, glucomannans and galactomannans. The enzyme has found a number of applications in different industries, including food, feed, pharmaceutical, pulp/paper industries, as well as gas well stimulation and pretreatment of lignocellulosic biomass for the production of second generation biofuel. Bacillus licheniformis is a Gram-positive endospore-forming microorganism that is generally non-pathogenic and has been used extensively for large-scale industrial production of various enzymes; however, there has been no previous report on the cloning and expression of mannan endo-1,4-beta-mannosidase gene (manB) from B. licheniformis. The mannan endo-1,4-beta-mannosidase gene (manB), commonly known as beta-mannanase, from Bacillus licheniformis strain DSM13 was cloned and overexpressed in Escherichia coli. The enzyme can be harvested from the cell lysate, periplasmic extract, or culture supernatant when using the pFLAG expression system. A total activity of approximately 50,000 units could be obtained from 1-l shake flask cultures. The recombinant enzyme was 6 x His-tagged at its C-terminus, and could be purified by one-step immobilized metal affinity chromatography (IMAC) to apparent homogeneity. The specific activity of the purified enzyme when using locust bean gum as substrate was 1672 +/- 96 units/mg. The optimal pH of the enzyme was between pH 6.0 - 7.0; whereas the optimal temperature was at 50 - 60 degrees C. The recombinant beta-mannanase was stable within pH 5 - 12 after incubation for 30 min at 50 degrees C, and within pH 6 - 9 after incubation at 50 degrees C for 24 h. The enzyme was stable at temperatures up to 50 degrees C with a half-life time of activity (tau1/2) of approximately 80 h at 50 degrees C and pH 6.0. Analysis of hydrolytic products by thin layer chromatography revealed that the main products from the bioconversion of locus bean gum and mannan were various manno-oligosaccharide products (M2 - M6) and mannose. Our study demonstrates an efficient expression and secretion system for the production of a relatively thermo- and alkali-stable recombinant beta-mannanase from B. licheniformis strain DSM13, suitable for various biotechnological applications.

  6. Research advances based on mass spectrometry for profiling of triacylglycerols in oils and fats and their applications.

    PubMed

    Xu, Shu-Ling; Wei, Fang; Xie, Ya; Lv, Xin; Dong, Xu-Yan; Chen, Hong

    2018-03-23

    Vegetable oils and animal fats are dietary source of lipids that play critical and multiple roles in biological function. Triacylglycerols (TAGs) are the principal component of oils and fats with significant difference in profile among different oils and fats. TAG profiling is essential for nutritional evaluation, quality control and assurance of safety in oils and fats. However, analysis of TAGs is a challenging task because of the complicated composition of TAGs and their similar physicochemical properties in oils and fats. The rapid development of mass spectrometry (MS) technology in recent years makes it possible to analyze the composition, content and structure of TAGs in the study of the physical, chemical and nutritional properties of oils, fats and related products. This review described the research advancement based on MS for profiling of TAGs in oil, fat and their applications in food. The application of MS, including direct infusion strategies, and its combination with chromatography, gas chromatography-MS (GC-MS) and liquid chromatography-MS (LC-MS), in the analysis of TAGs were reviewed. The advantages and disadvantages of these analytical methods with relevant applications for TAGs analysis in food were also described. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Design, synthesis, and application of the trimethoprim-based chemical tag for live-cell imaging.

    PubMed

    Jing, Chaoran; Cornish, Virginia W

    2013-01-01

    Over the past decade, chemical tags have been developed to complement the use of fluorescent proteins in live-cell imaging. Chemical tags retain the specificity of protein labeling achieved with fluorescent proteins through genetic encoding, but provide smaller, more robust tags and modular use of organic fluorophores with high photon output and tailored functionalities. The trimethoprim-based chemical tag (TMP-tag) was initially developed based on the high affinity interaction between E. coli dihydrofolate reductase and the antibiotic trimethoprim and was subsequently rendered covalent and fluorogenic via proximity-induced protein labeling reactions. To date, the TMP-tag is one of the few chemical tags that enable intracellular protein labeling and high-resolution live-cell imaging. Here we describe the general design, chemical synthesis, and application of TMP-tag for live-cell imaging. Alternate protocols for synthesizing and using the covalent and the fluorogenic TMP-tags are also included. © 2013 by John Wiley & Sons, Inc.

  8. Security Techniques for Prevention of Rank Manipulation in Social Tagging Services including Robotic Domains

    PubMed Central

    2014-01-01

    With smartphone distribution becoming common and robotic applications on the rise, social tagging services for various applications including robotic domains have advanced significantly. Though social tagging plays an important role when users are finding the exact information through web search, reliability and semantic relation between web contents and tags are not considered. Spams are making ill use of this aspect and put irrelevant tags deliberately on contents and induce users to advertise contents when they click items of search results. Therefore, this study proposes a detection method for tag-ranking manipulation to solve the problem of the existing methods which cannot guarantee the reliability of tagging. Similarity is measured for ranking the grade of registered tag on the contents, and weighted values of each tag are measured by means of synonym relevance, frequency, and semantic distances between tags. Lastly, experimental evaluation results are provided and its efficiency and accuracy are verified through them. PMID:25114975

  9. SAGE III/ISS L2 Solar Event Species Profiles (Native) V5 (g3bsspb)

    Atmospheric Science Data Center

    2017-12-21

    SAGE III/ISS L2 Solar Event Species Profiles (Native) V5 (g3bsspb)   Project ... present Temporal Resolution:  1 file per event File Format:  BINARY Tools:  Earthdata ... Radiation Longwave Radiation Shortwave Radiation Event Tag Event Type Obs Beta Angle Order Data:  ...

  10. SAGE III/ISS L2 Lunar Event Species Profiles (Native) V5 (g3blspb)

    Atmospheric Science Data Center

    2018-01-08

    SAGE III/ISS L2 Lunar Event Species Profiles (Native) V5 (g3blspb)   Project ... present Temporal Resolution:  1 file per event File Format:  BINARY Tools:  Earthdata ... Radiation Longwave Radiation Shortwave Radiation Event Tag Event Type Obs Beta Angle Order Data:  ...

  11. TeO$$_2$$ bolometers with Cherenkov signal tagging: towards next-generation neutrinoless double-beta decay experiments

    DOE PAGES

    Casali, N.; Vignati, Marco; Beeman, J. W.; ...

    2015-01-14

    CUORE, an array of 988 TeOmore » $$_2$$ bolometers, is about to be one of the most sensitive experiments searching for neutrinoless double-beta decay. Its sensitivity could be further improved by removing the background from α radioactivity. A few years ago it was pointed out that the signal from βs can be tagged by detecting the emitted Cherenkov light, which is not produced by αs. In this paper we confirm this possibility. For the first time we measured the Cherenkov light emitted by a CUORE crystal, and found it to be 100 eV at the Q-value of the decay. To completely reject the α background, we compute that one needs light detectors with baseline noise below 20 eV RMS, a value which is 3–4 times smaller than the average noise of the bolometric light detectors we are using. We point out that an improved light detector technology must be developed to obtain TeO$$_2$$ bolometric experiments able to probe the inverted hierarchy of neutrino masses.« less

  12. DOE Office of Scientific and Technical Information (OSTI.GOV)

    McDonald, A. D.; Jones, B. J. P.; Nygren, D. R.

    A new method to tag the barium daughter in the double beta decay ofmore » $$^{136}$$Xe is reported. Using the technique of single molecule fluorescent imaging (SMFI), individual barium dication (Ba$$^{++}$$) resolution at a transparent scanning surface has been demonstrated. A single-step photo-bleach confirms the single ion interpretation. Individual ions are localized with super-resolution ($$\\sim$$2~nm), and detected with a statistical significance of 12.9~$$\\sigma$$ over backgrounds. This lays the foundation for a new and potentially background-free neutrinoless double beta decay technology, based on SMFI coupled to high pressure xenon gas time projection chambers.« less

  13. Phospholipase D¿ enhances diacylglycerol flux into triacylglycerol

    USDA-ARS?s Scientific Manuscript database

    Plant seeds are the primary source of triacylglycerols (TAG) for food, feed, fuel, and industrial applications. As TAG is produced from diacylglycerol (DAG) successful engineering strategies to enhance TAG levels have focused on the conversion of DAG to TAG. However, the production of TAG can be lim...

  14. InkTag: Secure Applications on an Untrusted Operating System

    PubMed Central

    Hofmann, Owen S.; Kim, Sangman; Dunn, Alan M.; Lee, Michael Z.; Witchel, Emmett

    2014-01-01

    InkTag is a virtualization-based architecture that gives strong safety guarantees to high-assurance processes even in the presence of a malicious operating system. InkTag advances the state of the art in untrusted operating systems in both the design of its hypervisor and in the ability to run useful applications without trusting the operating system. We introduce paraverification, a technique that simplifies the InkTag hypervisor by forcing the untrusted operating system to participate in its own verification. Attribute-based access control allows trusted applications to create decentralized access control policies. InkTag is also the first system of its kind to ensure consistency between secure data and metadata, ensuring recoverability in the face of system crashes. PMID:24429939

  15. InkTag: Secure Applications on an Untrusted Operating System.

    PubMed

    Hofmann, Owen S; Kim, Sangman; Dunn, Alan M; Lee, Michael Z; Witchel, Emmett

    2013-01-01

    InkTag is a virtualization-based architecture that gives strong safety guarantees to high-assurance processes even in the presence of a malicious operating system. InkTag advances the state of the art in untrusted operating systems in both the design of its hypervisor and in the ability to run useful applications without trusting the operating system. We introduce paraverification , a technique that simplifies the InkTag hypervisor by forcing the untrusted operating system to participate in its own verification. Attribute-based access control allows trusted applications to create decentralized access control policies. InkTag is also the first system of its kind to ensure consistency between secure data and metadata, ensuring recoverability in the face of system crashes.

  16. Secure Integration of Radio Frequency Identification (RFID) Technology into a Supply Chain

    DTIC Science & Technology

    2005-09-01

    serves as the rough equivalent of a license plate on an automobile . Figure 1 (below) illustrates the typical construction of an RFID tag. An antenna...writable passive tags (RW) Reprogrammable Class 3 Semi-active tags Reprogrammable Class 4 Active tags Reprogrammable Class 5 Readers... Reprogrammable Table 1. EPC Tag Classes[3]. Table 2 summarizes the advantages, disadvantages and applications of each type of tag. Tag Type Advantages

  17. Barium Tagging n Solid Xenon for nEXO Neutrinoless Double Beta Decay

    NASA Astrophysics Data System (ADS)

    Walton, Tim; Chambers, Chris; Craycraft, Adam; Fairbank, William; nEXO Collaboration

    2015-04-01

    nEXO is a next-generation experiment designed to search for neutrinoless double beta decay of the isotope Xe136 in a liquid xenon time projection chamber. Positive observation of this decay would determine the nature of the neutrino to be a Majorana particle. Since the daughter of this decay is barium (Ba136), detecting the presence of Ba136 at a decay site (called ``barium tagging'') would provide strong rejection of backgrounds in the search for this decay. This would involve detecting a single barium ion from within a macroscopic volume of liquid xenon. This technique may be available for a second phase of the nEXO detector and sensitivity beyond the inverted hierarchy to neutrino oscillations. Several methods of barium tagging are being explored by the nEXO collaboration, but here we present a method of trapping the barium ion/atom (it may neutralize) in solid xenon (SXe) at the end of a cold probe, and then detecting the ion/atom by its fluorescence in the SXe. Our group at CSU has been studying the fluorescence of Ba in SXe by laser excitation, in order to ultimately detect a single Ba +/Ba in a SXe sample. We present studies of fluorescence signals, as well as recent results on imaging small numbers of Ba atoms in SXe, in a focused laser region. This work is supported by grants from the National Science Foundation and the Department of Energy.

  18. RFID-Based Vehicle Positioning and Its Applications in Connected Vehicles

    PubMed Central

    Wang, Jianqiang; Ni, Daiheng; Li, Keqiang

    2014-01-01

    This paper proposed an RFID-based vehicle positioning approach to facilitate connected vehicles applications. When a vehicle passes over an RFID tag, the vehicle position is given by the accurate position stored in the tag. At locations without RFID coverage, the vehicle position is estimated from the most recent tag location using a kinematics integration algorithm till updates from the next tag. The accuracy of RFID positioning is verified empirically in two independent ways with one using radar and the other a photoelectric switch. The former is designed to verify whether the dynamic position obtained from RFID tags matches the position measured by radar that is regarded as accurate. The latter aims to verify whether the position estimated from the kinematics integration matches the position obtained from RFID tags. Both means supports the accuracy of RFID-based positioning. As a supplement to GPS which suffers from issues such as inaccuracy and loss of signal, RFID positioning is promising in facilitating connected vehicles applications. Two conceptual applications are provided here with one in vehicle operational control and the other in Level IV intersection control. PMID:24599188

  19. RFID-based vehicle positioning and its applications in connected vehicles.

    PubMed

    Wang, Jianqiang; Ni, Daiheng; Li, Keqiang

    2014-03-04

    This paper proposed an RFID-based vehicle positioning approach to facilitate connected vehicles applications. When a vehicle passes over an RFID tag, the vehicle position is given by the accurate position stored in the tag. At locations without RFID coverage, the vehicle position is estimated from the most recent tag location using a kinematics integration algorithm till updates from the next tag. The accuracy of RFID positioning is verified empirically in two independent ways with one using radar and the other a photoelectric switch. The former is designed to verify whether the dynamic position obtained from RFID tags matches the position measured by radar that is regarded as accurate. The latter aims to verify whether the position estimated from the kinematics integration matches the position obtained from RFID tags. Both means supports the accuracy of RFID-based positioning. As a supplement to GPS which suffers from issues such as inaccuracy and loss of signal, RFID positioning is promising in facilitating connected vehicles applications. Two conceptual applications are provided here with one in vehicle operational control and the other in Level IV intersection control.

  20. The alpha/beta hydrolase CGI-58 and peroxisomal transport protein PXA1 coregulate lipid homeostasis and signaling in Arabidopsis

    USDA-ARS?s Scientific Manuscript database

    COMPARATIVE GENE IDENTIFICATION-58 (CGI-58) is a key regulator of lipid metabolism and signaling in mammals, but its underlying mechanisms are unclear. Disruption of CGI-58 in either mammals or plants results in a significant increase in triacylglycerol (TAG), suggesting that CGI-58 activity is evol...

  1. SAGE III/ISS L1B Solar Event Transmission Data (Native) V5 (g3btb)

    Atmospheric Science Data Center

    2017-12-21

    SAGE III/ISS L1B Solar Event Transmission Data (Native) V5 (g3btb)   Project Title:  ... present Temporal Resolution:  1 file per event File Format:  BINARY Tools:  Earthdata ... Radiation Longwave Radiation Shortwave Radiation Event Tag Event Type Obs Beta Angle Order Data:  ...

  2. SAGE III/ISS L2 Solar Event Species Profiles (HDF-EOS) V5 (g3bssp)

    Atmospheric Science Data Center

    2017-12-21

    SAGE III/ISS L2 Solar Event Species Profiles (HDF-EOS) V5 (g3bssp)   Project ... present Temporal Resolution:  1 file per event File Format:  HDF-4 Tools:  Earthdata ... Radiation Longwave Radiation Shortwave Radiation Event Tag Event Type Obs Beta Angle Order Data:  ...

  3. SAGE III/ISS L2 Lunar Event Species Profiles (HDF-EOS) V5 (g3blsp)

    Atmospheric Science Data Center

    2018-01-04

    SAGE III/ISS L2 Lunar Event Species Profiles (HDF-EOS) V5 (g3blsp)   Project ... present Temporal Resolution:  1 file per event File Format:  HDF-4 Tools:  Earthdata ... Radiation Longwave Radiation Shortwave Radiation Event Tag Event Type Obs Beta Angle Order Data:  ...

  4. Use of a purified and functional recombinant calcium-channel beta4 subunit in surface-plasmon resonance studies.

    PubMed Central

    Geib, Sandrine; Sandoz, Guillaume; Mabrouk, Kamel; Matavel, Alessandra; Marchot, Pascale; Hoshi, Toshinori; Villaz, Michel; Ronjat, Michel; Miquelis, Raymond; Lévêque, Christian; de Waard, Michel

    2002-01-01

    Native high-voltage-gated calcium channels are multi-subunit complexes comprising a pore-forming subunit Ca(v) and at least two auxiliary subunits alpha(2)delta and beta. The beta subunit facilitates cell-surface expression of the channel and contributes significantly to its biophysical properties. In spite of its importance, detailed structural and functional studies are hampered by the limited availability of native beta subunit. Here, we report the purification of a recombinant calcium-channel beta(4) subunit from bacterial extracts by using a polyhistidine tag. The purified protein is fully functional since it binds on the alpha1 interaction domain, its main Ca(v)-binding site, and regulates the activity of P/Q calcium channel expressed in Xenopus oocytes in a similar way to the beta(4) subunit produced by cRNA injection. We took advantage of the functionality of the purified material to (i) develop an efficient surface-plasmon resonance assay of the interaction between two calcium channel subunits and (ii) measure, for the first time, the affinity of the recombinant His-beta(4) subunit for the full-length Ca(v)2.1 channel. The availability of this purified material and the development of a surface-plasmon resonance assay opens two immediate research perspectives: (i) drug screening programmes applied to the Ca(v)/beta interaction and (ii) crystallographic studies of the calcium-channel beta(4) subunit. PMID:11988102

  5. Hypertrophic stimulation increases beta-actin dynamics in adult feline cardiomyocytes.

    PubMed

    Balasubramanian, Sundaravadivel; Mani, Santhosh K; Kasiganesan, Harinath; Baicu, Catalin C; Kuppuswamy, Dhandapani

    2010-07-12

    The myocardium responds to hemodynamic stress through cellular growth and organ hypertrophy. The impact of cytoskeletal elements on this process, however, is not fully understood. While alpha-actin in cardiomyocytes governs muscle contraction in combination with the myosin motor, the exact role of beta-actin has not been established. We hypothesized that in adult cardiomyocytes, as in non-myocytes, beta-actin can facilitate cytoskeletal rearrangement within cytoskeletal structures such as Z-discs. Using a feline right ventricular pressure overload (RVPO) model, we measured the level and distribution of beta-actin in normal and pressure overloaded myocardium. Resulting data demonstrated enriched levels of beta-actin and enhanced translocation to the Triton-insoluble cytoskeletal and membrane skeletal complexes. In addition, RVPO in vivo and in vitro hypertrophic stimulation with endothelin (ET) or insulin in isolated adult cardiomyocytes enhanced the content of polymerized fraction (F-actin) of beta-actin. To determine the localization and dynamics of beta-actin, we adenovirally expressed GFP-tagged beta-actin in isolated adult cardiomyocytes. The ectopically expressed beta-actin-GFP localized to the Z-discs, costameres, and cell termini. Fluorescence recovery after photobleaching (FRAP) measurements of beta-actin dynamics revealed that beta-actin at the Z-discs is constantly being exchanged with beta-actin from cytoplasmic pools and that this exchange is faster upon hypertrophic stimulation with ET or insulin. In addition, in electrically stimulated isolated adult cardiomyocytes, while beta-actin overexpression improved cardiomyocyte contractility, immunoneutralization of beta-actin resulted in a reduced contractility suggesting that beta-actin could be important for the contractile function of adult cardiomyocytes. These studies demonstrate the presence and dynamics of beta-actin in the adult cardiomyocyte and reinforce its usefulness in measuring cardiac cytoskeletal rearrangement during hypertrophic stimulation.

  6. Characterisation and confirmation of rare beta-thalassaemia mutations in the Malay, Chinese and Indian ethnic groups in Malaysia.

    PubMed

    Tan, Jin Ai Mary Anne; Chin, Pui See; Wong, Yean Ching; Tan, Kim Lian; Chan, Lee Lee; George, Elizabeth

    2006-10-01

    In Malaysia, about 4.5% of the Malay and Chinese populations are heterozygous carriers of beta-thalassaemia. The initial identification of rare beta-globin gene mutations by genomic sequencing will allow the development of simpler and cost-effective PCR-based techniques to complement the existing amplification refractory mutation system (ARMS) and gap-PCR used for the identification of beta-thalassaemia mutations. DNA from 173 beta-thalassaemia carriers and five beta-thalassaemia major patients from the Malay, Chinese and Indian ethnic groups were first analysed by ARMS and gap-PCR. Ninety-five per cent (174/183) of the 183 beta-globin genes studied were characterised using these two techiques. The remaining nine uncharacterised beta-globin genes (4.9%) were analysed using genomic sequencing of a 904 bp amplified PCR product consisting of the promoter region, exon 1, intervening sequence (IVS) 1, exon 2 and the 5' IVS2 regions of the beta-globin gene. The rare beta-globin mutations detected in the Chinese patients were CD27/28 (+C) and CD43 (GAG-TAG), and -88 (C-T) in an Indian patient. Beta-globin mutations at CD16 (-C), IVS1-1 (G-A), IVS2-1 (G-A), -86 (C-G) and Haemoglobin South Florida (CD1, GTG-ATG) were confirmed in the Malay patients. The seven rare beta-globin mutations and a rare haemoglobin variant confirmed in this study have been described in other populations but have not been previously described in Malaysian beta-thalassemia patients.

  7. Molecular phenotype of zebrafish ovarian follicle by serial analysis of gene expression and proteomic profiling, and comparison with the transcriptomes of other animals

    PubMed Central

    Knoll-Gellida, Anja; André, Michèle; Gattegno, Tamar; Forgue, Jean; Admon, Arie; Babin, Patrick J

    2006-01-01

    Background The ability of an oocyte to develop into a viable embryo depends on the accumulation of specific maternal information and molecules, such as RNAs and proteins. A serial analysis of gene expression (SAGE) was carried out in parallel with proteomic analysis on fully-grown ovarian follicles from zebrafish (Danio rerio). The data obtained were compared with ovary/follicle/egg molecular phenotypes of other animals, published or available in public sequence databases. Results Sequencing of 27,486 SAGE tags identified 11,399 different ones, including 3,329 tags with an occurrence superior to one. Fifty-eight genes were expressed at over 0.15% of the total population and represented 17.34% of the mRNA population identified. The three most expressed transcripts were a rhamnose-binding lectin, beta-actin 2, and a transcribed locus similar to the H2B histone family. Comparison with the large-scale expressed sequence tags sequencing approach revealed highly expressed transcripts that were not previously known to be expressed at high levels in fish ovaries, like the short-sized polarized metallothionein 2 transcript. A higher sensitivity for the detection of transcripts with a characterized maternal genetic contribution was also demonstrated compared to large-scale sequencing of cDNA libraries. Ferritin heavy polypeptide 1, heat shock protein 90-beta, lactate dehydrogenase B4, beta-actin isoforms, tubulin beta 2, ATP synthase subunit 9, together with 40 S ribosomal protein S27a, were common highly-expressed transcripts of vertebrate ovary/unfertilized egg. Comparison of transcriptome and proteome data revealed that transcript levels provide little predictive value with respect to the extent of protein abundance. All the proteins identified by proteomic analysis of fully-grown zebrafish follicles had at least one transcript counterpart, with two exceptions: eosinophil chemotactic cytokine and nothepsin. Conclusion This study provides a complete sequence data set of maternal mRNA stored in zebrafish germ cells at the end of oogenesis. This catalogue contains highly-expressed transcripts that are part of a vertebrate ovarian expressed gene signature. Comparison of transcriptome and proteome data identified downregulated transcripts or proteins potentially incorporated in the oocyte by endocytosis. The molecular phenotype described provides groundwork for future experimental approaches aimed at identifying functionally important stored maternal transcripts and proteins involved in oogenesis and early stages of embryo development. PMID:16526958

  8. Characterization of cellulolytic activity from digestive fluids of Dissosteira carolina (Orthoptera: Acrididae).

    PubMed

    Willis, Jonathan D; Klingeman, William E; Oppert, Cris; Oppert, Brenda; Jurat-Fuentes, Juan L

    2010-11-01

    Previous screening of head-derived and gut fluid extracts of Carolina grasshoppers, Dissosteira carolina (L.) revealed relatively high activity against cellulase substrates when compared to other insect groups. In this work we report on the characterization and identification of enzymes involved in cellulolytic activity in digestive fluids of D. carolina. In zymograms using carboxymethylcellulose (CMC) as substrate, we detected four distinct cellulolytic protein bands in D. carolina gut fluids, common to all developmental stages. These cellulolytic enzymes were localized to foregut and midgut regions of the D. carolina digestive tract. Cellulases were purified from D. carolina head and gut fluid extracts by liquid chromatography to obtain N-terminal amino acid sequence tags. Database searches with sequence tags from head fluids indicated high similarity with invertebrate, bacterial and plant beta1,4-endoglucanases, while no homologues were identified for the gut-derived protein. Our data demonstrate the presence of cellulolytic activity in the digestive system of D. carolina and suggest that cellulases of endogenous origin are present in this organism. Considering that this grasshopper species is a pest of grasses, including switchgrass that has been suggested bioethanol feedstock, characterization of insect cellulolytic systems may aid in developing applications for plant biomass biodegradation for biofuel production. Copyright 2010 Elsevier Inc. All rights reserved.

  9. The 17 beta-oestradiol dehydrogenase of pig endometrial cells is localized in specialized vesicles.

    PubMed Central

    Adamski, J; Husen, B; Marks, F; Jungblut, P W

    1993-01-01

    Two monoclonal antibodies against the 17 beta-oestradiol dehydrogenase of pig endometrial cells have been used in localization studies with immunogold electron microscopy. The antibodies attach both to a fraction of dehydrogenase-rich cytoplasmic vesicles isolated from homogenates and to vesicles of similar appearance in cells. The vesicles are filled with electron-dense material. Their tagging intensity indicates a high degree of specialization. Endometrial cells from mature animals contain a host of dehydrogenase vesicles, and cells from prepubertal animals only a few. Functional aspects of the novel organelle are discussed. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:8457206

  10. Barium Tagging from nEXO Using Resonance Ionization Spectroscopy

    NASA Astrophysics Data System (ADS)

    Twelker, K.; Kravitz, S.

    nEXO is a 5-ton liquid enriched-xenon time projection chamber (TPC) to search for neutrinoless double-beta decay, designed to have the sensitivity to completely probe the inverted mass hierarchy of Majorana neutrinos. The detector will accommodate-as a background reduction technique-a system to recover and identify the barium decay product. This upgrade will allow a background-free measurement of neutrinoless double-beta decay and increase the half-life sensitivity of the experiment by at least one order of magnitude. Ongoing research and development includes a system to test barium extraction from liquid xenon using surface adsorption and Resonance Ionization Spectroscopy (RIS).

  11. Cherenkov light identification in TeO2 crystals with Si low-temperature detectors

    NASA Astrophysics Data System (ADS)

    Gironi, L.; Biassoni, M.; Brofferio, C.; Capelli, S.; Carniti, P.; Cassina, L.; Clemenza, M.; Cremonesi, O.; Faverzani, M.; Ferri, E.; Giachero, A.; Gotti, C.; Maino, M.; Margesin, B.; Nucciotti, A.; Pavan, M.; Pessina, G.; Pozzi, S.; Previtali, E.; Puiu, A.; Sisti, M.; Terranova, F.

    2017-09-01

    Low temperature thermal detectors with particle identification capabilities are among the best detectors for next generation experiments for the search of neutrinoless double beta decay. Thermal detectors allow to reach excellent energy resolution and to optimize the detection efficiency, while the possibility to identify the interacting particle allows to greatly reduce the background. Tellurium dioxide is one of the favourite compounds since it has long demonstrated the first two features and could reach the third through Cherenkov emission tagging [1]. A new generation of cryogenic light detectors are however required to detect the few Cherenkov photons emitted by electrons of few MeV energy. Preliminary measurements with new Si light detectors demonstrated a clear event-by-event discrimination between alpha and beta/gamma interactions at the 130Te neutrinoless double beta decay Q-value (2528 keV).

  12. SAGE III/ISS L1B Solar Event Transmission Data (HDF-EOS) V5 (g3bt)

    Atmospheric Science Data Center

    2017-12-21

    SAGE III/ISS L1B Solar Event Transmission Data (HDF-EOS) V5 (g3bt)   Project Title:  ... present Temporal Resolution:  1 file per event File Format:  HDF-4 Tools:  Earthdata ... Radiation Longwave Radiation Shortwave Radiation Event Tag Event Type Obs Beta Angle Order Data:  ...

  13. Application safety evaluation of the radio frequency identification tag under magnetic resonance imaging.

    PubMed

    Fei, Xiaolu; Li, Shanshan; Gao, Shan; Wei, Lan; Wang, Lihong

    2014-09-04

    Radio Frequency Identification(RFID) has been widely used in healthcare facilities, but it has been paid little attention whether RFID applications are safe enough under healthcare environment. The purpose of this study is to assess the effects of RFID tags on Magnetic Resonance (MR) imaging in a typical electromagnetic environment in hospitals, and to evaluate the safety of their applications. A Magphan phantom was used to simulate the imaging objects, while active RFID tags were placed at different distances (0, 4, 8, 10 cm) from the phantom border. The phantom was scanned by using three typical sequences including spin-echo (SE) sequence, gradient-echo (GRE) sequence and inversion-recovery (IR) sequence. The quality of the image was quantitatively evaluated by using signal-to-noise ratio (SNR), uniformity, high-contrast resolution, and geometric distortion. RFID tags were read by an RFID reader to calculate their usable rate. RFID tags can be read properly after being placed in high magnetic field for up to 30 minutes. SNR: There were no differences between the group with RFID tags and the group without RFID tags using SE and IR sequence, but it was lower when using GRE sequence.Uniformity: There was a significant difference between the group with RFID tags and the group without RFID tags using SE and GRE sequence. Geometric distortion and high-contrast resolution: There were no obvious differences found. Active RFID tags can affect MR imaging quality, especially using the GRE sequence. Increasing the distance from the RFID tags to the imaging objects can reduce that influence. When the distance was longer than 8 cm, MR imaging quality were almost unaffected. However, the Gradient Echo related sequence is not recommended when patients wear a RFID wristband.

  14. Total absorption spectroscopy of fission fragments relevant for reactor antineutrino spectra

    DOE PAGES

    Fallot, M.; Porta, A.; Meur, L. Le; ...

    2017-09-13

    Here, the accurate determination of reactor antineutrino spectra remains a very active research topic for which new methods of study have emerged in recent years. Indeed, following the long-recognized reactor anomaly (measured antineutrino deficit in short baseline reactor experiments when compared with spectral predictions), the three international reactor neutrino experiments Double Chooz, Daya Bay and Reno have recently demonstrated the existence of spectral distortions in their measurements with respect to the same predictions. These spectral predictions were obtained through the conversion of integral beta-energy spectra obtained at the ILL research reactor. Several studies have shown that the underlying nuclear physicsmore » required for the conversion of these spectra into antineutrino spectra is not totally understood. An alternative to such converted spectra is a complementary approach that consists of determining the antineutrino spectrum by means of the measurement and processing of nuclear data. The beta properties of some key fission products suffer from the pandemonium effect which can be circumvented by the use of the Total Absorption Gamma-ray Spectroscopy technique (TAGS). The two main contributors to the Pressurized Water Reactor antineutrino spectrum in the region where the spectral distortion has been observed are 92Rb and 142Cs, which have been measured at the radioactive beam facility of the University of Jyvaskyla in two TAGS experiments. We present the results of the analysis of the TAGS measurements of the β-decay properties of 92Rb along with preliminary results on 142Cs and report on the measurements already performed.« less

  15. Total absorption spectroscopy of fission fragments relevant for reactor antineutrino spectra

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fallot, M.; Porta, A.; Meur, L. Le

    Here, the accurate determination of reactor antineutrino spectra remains a very active research topic for which new methods of study have emerged in recent years. Indeed, following the long-recognized reactor anomaly (measured antineutrino deficit in short baseline reactor experiments when compared with spectral predictions), the three international reactor neutrino experiments Double Chooz, Daya Bay and Reno have recently demonstrated the existence of spectral distortions in their measurements with respect to the same predictions. These spectral predictions were obtained through the conversion of integral beta-energy spectra obtained at the ILL research reactor. Several studies have shown that the underlying nuclear physicsmore » required for the conversion of these spectra into antineutrino spectra is not totally understood. An alternative to such converted spectra is a complementary approach that consists of determining the antineutrino spectrum by means of the measurement and processing of nuclear data. The beta properties of some key fission products suffer from the pandemonium effect which can be circumvented by the use of the Total Absorption Gamma-ray Spectroscopy technique (TAGS). The two main contributors to the Pressurized Water Reactor antineutrino spectrum in the region where the spectral distortion has been observed are 92Rb and 142Cs, which have been measured at the radioactive beam facility of the University of Jyvaskyla in two TAGS experiments. We present the results of the analysis of the TAGS measurements of the β-decay properties of 92Rb along with preliminary results on 142Cs and report on the measurements already performed.« less

  16. Targeted Multiplex Imaging Mass Spectrometry in Transmission Geometry for Subcellular Spatial Resolution

    PubMed Central

    Lavenant, Gwendoline Thiery; Zavalin, Andrey I.; Caprioli, Richard M.

    2013-01-01

    Targeted multiplex Imaging Mass Spectrometry utilizes several different antigen-specific primary antibodies, each directly labeled with a unique photocleavable mass tag, to detect multiple antigens in a single tissue section. Each photocleavable mass tag bound to an antibody has a unique molecular weight and can be readily ionized by laser desorption ionization mass spectrometry. This manuscript describes a mass spectrometry method that allows imaging of targeted single cells within tissue using transmission geometry laser desorption ionization mass spectrometry. Transmission geometry focuses the laser beam on the back side of the tissue placed on a glass slide, providing a 2 μm diameter laser spot irradiating the biological specimen. This matrix-free method enables simultaneous localization at the sub-cellular level of multiple antigens using specific tagged antibodies. We have used this technology to visualize the co-expression of synaptophysin and two major hormones peptides, insulin and somatostatin, in duplex assays in beta and delta cells contained in a human pancreatic islet. PMID:23397138

  17. An internal affinity-tag for purification and crystallization of the siderophore receptor FhuA, integral outer membrane protein from Escherichia coli K-12.

    PubMed

    Ferguson, A D; Breed, J; Diederichs, K; Welte, W; Coulton, J W

    1998-07-01

    FhuA (Mr 78,992, 714 amino acids), siderophore receptor for ferrichrome-iron in the outer membrane of Escherichia coli, was affinity tagged, rapidly purified, and crystallized. To obtain FhuA in quantities sufficient for crystallization, a hexahistidine tag was genetically inserted into the fhuA gene after amino acid 405, which resides in a known surface-exposed loop. Recombinant FhuA405.H6 was overexpressed in an E. coli strain that is devoid of several major porins and using metal-chelate chromatography was purified in large amounts to homogeneity. FhuA crystals were grown using the hanging drop vapor diffusion technique and were suitable for X-ray diffraction analysis. On a rotating anode X-ray source, diffraction was observed to 3.0 A resolution. The crystals belong to space group P6(1) or P6(5) with unit cell dimensions of a=b=174 A, c=88 A (alpha=beta=90 degrees, gamma=120 degrees).

  18. Targeted Multiplex Imaging Mass Spectrometry in Transmission Geometry for Subcellular Spatial Resolution

    NASA Astrophysics Data System (ADS)

    Thiery-Lavenant, Gwendoline; Zavalin, Andre I.; Caprioli, Richard M.

    2013-04-01

    Targeted multiplex imaging mass spectrometry utilizes several different antigen-specific primary antibodies, each directly labeled with a unique photocleavable mass tag, to detect multiple antigens in a single tissue section. Each photocleavable mass tag bound to an antibody has a unique molecular weight and can be readily ionized by laser desorption ionization mass spectrometry. This article describes a mass spectrometry method that allows imaging of targeted single cells within tissue using transmission geometry laser desorption ionization mass spectrometry. Transmission geometry focuses the laser beam on the back side of the tissue placed on a glass slide, providing a 2 μm diameter laser spot irradiating the biological specimen. This matrix-free method enables simultaneous localization at the sub-cellular level of multiple antigens using specific tagged antibodies. We have used this technology to visualize the co-expression of synaptophysin and two major hormones peptides, insulin and somatostatin, in duplex assays in beta and delta cells contained in a human pancreatic islet.

  19. Anti-collision radio-frequency identification system using passive SAW tags

    NASA Astrophysics Data System (ADS)

    Sorokin, A. V.; Shepeta, A. P.

    2017-06-01

    Modern multi sensor systems should have high operating speed and resistance to climate impacts. Radiofrequency systems use passive SAW tags for identification items and vehicles. These tags find application in industry, traffic remote control systems, and railway remote traffic control systems for identification and speed measuring. However, collision of the passive SAW RFID tags hinders development passive RFID SAW technology in Industry. The collision problem for passive SAW tags leads for incorrect identification and encoding each tag. In our researching, we suggest approach for identification of several passive SAW tags in collision case.

  20. iTAG: Integrating a Cloud Based, Collaborative Animal Tracking Network into the GCOOS data portal in the Gulf of Mexico

    NASA Astrophysics Data System (ADS)

    Kirkpatrick, B. A.; Currier, R. D.; Simoniello, C.

    2016-02-01

    The tagging and tracking of aquatic animals using acoustic telemetry hardware has traditionally been the purview of individual researchers that specialize in single species. Concerns over data privacy and unauthorized use of receiver arrays have prevented the construction of large-scale, multi-species, multi-institution, multi-researcher collaborative acoustic arrays. We have developed a toolset to build the new portal using the Flask microframework, Python language, and Twitter bootstrap. Initial feedback has been overwhelmingly positive. The privacy policy has been praised for its granularity: principal investigators can choose between three levels of privacy for all data and hardware: Completely private - viewable only by the PI Visible to iTAG members Visible to the general public At the time of this writing iTAG is still in the beta stage, but the feedback received to date indicates that with the proper design and security features, and an iterative cycle of feedback from potential members, constructing a collaborative acoustic tracking network system is possible. Initial usage will be limited to the entry and searching for `orphan/mystery' tags, with the integration of historical array deployments and data following shortly thereafter. We have also been working with staff from the Ocean Tracking Network to allow for integration of the two systems. The database schema of iTAG is based on the marine metadata convention for acoustic telemetry. This should permit machine-to-machine data exchange between iTAG and OTN. The integration of animal telemetry data into the GCOOS portal will allow researchers to easily access the physiochemical oceanography data, thus allowing for a more in depth understanding of animal response and usage patterns.

  1. Reduction of high-affinity beta2-adrenergic receptor binding by hyperforin and hyperoside on rat C6 glioblastoma cells measured by fluorescence correlation spectroscopy.

    PubMed

    Prenner, Lars; Sieben, Anne; Zeller, Karin; Weiser, Dieter; Häberlein, Hanns

    2007-05-01

    Beta-adrenergic receptors (beta-AR) are potential targets for antidepressants. Desensitization and downregulation of beta-AR are discussed as possible modes of action for antidepressants. We have investigated the effects of hyperforin and hyperoside, compounds with potentially antidepressant activity from St. John's Wort, on the binding behavior and dynamics of beta2-AR in living rat C6 glioblastoma cells, compared to desipramine (desmethylimipramine; DMI) by means of fluorescence correlation spectroscopy (FCS) and fluorescence microscopy. FCS-binding studies with the fluorescently labeled ligand Alexa532-noradrenaline (Alexa532-NA) binding to beta2-AR of C6 cells showed a significant reduction in total beta2-AR binding after preincubation with hyperforin and hyperoside for 3 days, respectively, which was also found for DMI. This was mainly observed in high-affinity receptor-ligand complexes with hindered lateral mobility (D2 = 1.1 (+/-0.4) microm2/s) in the biomembrane. However, internalization of beta2-AR was found neither in z-scans of these C6 cells nor in HEK 293 cells stably transfected with GFP-tagged beta2-adrenergic receptors (beta2AR-GFP) after incubation up to 6 days with either DMI, hyperforin, or hyperoside. Thus, under these conditions reduction of beta2-AR binding was not mediated by receptor internalization. Additionally, preincubation of C6 cells with DMI, hyperforin, and hyperoside led to a loss of second messenger cAMP after beta2-adrenergic stimulating conditions with terbutaline. Our current results indicate that hyperforin and hyperoside from St. John's Wort, as well as DMI, reduce beta2-adrenergic sensitivity in C6 cells, emphasizing the potential usefulness of St. John's Wort dry extracts in clinical treatment of depressive symptoms.

  2. Radon-related Backgrounds in the LUX Dark Matter Search

    NASA Astrophysics Data System (ADS)

    Bradley, A.; Akerib, D. S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Bernard, E.; Bernstein, A.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chapman, J. J.; Chiller, A. A.; Chiller, C.; Coffey, T.; Currie, A.; de Viveiros, L.; Dobi, A.; Dobson, J.; Druszkiewicz, E.; Edwards, B.; Faham, C. H.; Fiorucci, S.; Flores, C.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C.; Hertel, S. A.; Horn, M.; Huang, D. Q.; Ihm, M.; Jacobsen, R. G.; Kazkaz, K.; Knoche, R.; Larsen, N. A.; Lee, C.; Lindote, A.; Lopes, M. I.; Malling, D. C.; Mannino, R.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H.; Neves, F.; Ott, R. A.; Pangilinan, M.; Parker, P. D.; Pease, E. K.; Pech, K.; Phelps, P.; Reichhart, L.; Shutt, T.; Silva, C.; Solovov, V. N.; Sorensen, P.; O'Sullivan, K.; Sumner, T. J.; Szydagis, M.; Taylor, D.; Tennyson, B.; Tiedt, D. R.; Tripathi, M.; Uvarov, S.; Verbus, J. R.; Walsh, N.; Webb, R.; White, J. T.; Witherell, M. S.; Wolfs, F. L. H.; Woods, M.; Zhang, C.

    The LUX detector is currently in operation at the Davis Campus at the 4850' level of the Sanford Underground Research Facility (SURF) in Lead, SD to directly search for WIMP dark matter. Knowing the type and rate of backgrounds is critical in a rare, low energy event search, and LUX was designed, constructed, and deployed to mitigate backgrounds, both internal and external. An important internal background are decays of radon and its daughters. These consist of alpha decays, which are easily tagged and are a tracer of certain backgrounds, and beta decays, some of which are not as readily tagged and present a background for the WIMP search. We report on studies of alpha decay and discuss implications for the WIMP search.

  3. Design and testing of RFID sensor tag fabricated using inkjet-printing and electrodeposition

    NASA Astrophysics Data System (ADS)

    Chien Dang, Mau; Son Nguyen, Dat; Dung Dang, Thi My; Tedjini, Smail; Fribourg-Blanc, Eric

    2014-06-01

    The passive RFID tag with an added sensing function is of interest to many applications. In particular, applications where RFID tagging is already considered to be the next step, such as food items, are a specific target. This paper demonstrates a flexible RFID tag sensor fabricated using a low cost technique with an added zero-cost sensing function. It is more specifically applied to the sensing of degradable food, in particular beef meat in our demonstrated example. To reach this, the antenna is designed in such a way to be sensitive to the variation of the dielectric permittivity of the meat over time. The design of the sensing tag as well as its fabrication process are described. The fabrication involves inkjet printing of a silver nanoparticle based ink on a commercial low cost PET film to create a seed layer. It is followed by a copper electrodeposition step on top of the silver pattern to complete the tag to obtain the desired thickness and conductivity of the tag antenna. The results of the electrical tests showed that with the inkjet printing-electrodeposition combination it is possible to produce flexible electrically conductive patterns for practical RFID applications. The tag was then tested in close-to-real-world conditions and it is demonstrated that it can provide a sensing function to detect the consumption limit of the packaged beef.

  4. A universal TagModule collection for parallel genetic analysis of microorganisms

    PubMed Central

    Oh, Julia; Fung, Eula; Price, Morgan N.; Dehal, Paramvir S.; Davis, Ronald W.; Giaever, Guri; Nislow, Corey; Arkin, Adam P.; Deutschbauer, Adam

    2010-01-01

    Systems-level analyses of non-model microorganisms are limited by the existence of numerous uncharacterized genes and a corresponding over-reliance on automated computational annotations. One solution to this challenge is to disrupt gene function using DNA tag technology, which has been highly successful in parallelizing reverse genetics in Saccharomyces cerevisiae and has led to discoveries in gene function, genetic interactions and drug mechanism of action. To extend the yeast DNA tag methodology to a wide variety of microorganisms and applications, we have created a universal, sequence-verified TagModule collection. A hallmark of the 4280 TagModules is that they are cloned into a Gateway entry vector, thus facilitating rapid transfer to any compatible genetic system. Here, we describe the application of the TagModules to rapidly generate tagged mutants by transposon mutagenesis in the metal-reducing bacterium Shewanella oneidensis MR-1 and the pathogenic yeast Candida albicans. Our results demonstrate the optimal hybridization properties of the TagModule collection, the flexibility in applying the strategy to diverse microorganisms and the biological insights that can be gained from fitness profiling tagged mutant collections. The publicly available TagModule collection is a platform-independent resource for the functional genomics of a wide range of microbial systems in the post-genome era. PMID:20494978

  5. Ultrasound-assisted synthesis of santalbic acid and a study of triacylglycerol species in Santalum album (Linn.) seed oil.

    PubMed

    Lie Ken Jie, M S; Pasha, M K; Ahmad, F

    1996-10-01

    Methyl ricinoleate (1) was treated with bromine and the dibromo derivative (2) was reacted with ethanolic KOH under ultrasonic irradiation to give 12-hydroxy-octadec-9-ynoic acid upon acidification with dil. HCI. The latter compound was methylated with BF3/methanol to give methyl 12-hydroxy-octadec-9-ynoate (3). Compound 3 was treated with methanesulfonyl chloride in the presence of triethylamine in CH2Cl2 to give methyl 12-mesyloxy-octadec-9-ynoate (4). Reaction of methyl 12-mesyloxy-octadec-9-ynoate with aqueous KOH under ultrasonic irradiation (20 kHz) gave (11E)-octadecen-9-ynoic acid (5, santalbic acid, 40%) and (11Z)-octadecen-9-ynoic acid (6, 60%) on acidification with dil. HCI. These isomers were separated by urea fractionation. The 13C nuclear magnetic resonance (NMR) spectroscopic properties of the methyl ester and the triacylglycerol (TAG) esters of these enynoic fatty acid isomers were studied. The carbon shifts of the unsaturated carbon nuclei of the methyl ester of the E-isomer were unambiguously assigned as 88.547 (C-9), 79.287 (C-10), 109.760 (C-11), and 143.450 (C-12) ppm, while the unsaturated carbon shifts of the (Z)-enynoate isomer appeared at 94.277 (C-9), 77.561 (C-10), 109.297 (C-11), and 142.668 (C-12) ppm. In the 13C NMR spectral analysis of the TAG molecules of type AAA containing either the (Z)- or (E)-enyne fatty acid, the C-1 to C-6 carbon atoms on the alpha- and beta-acyl positions were differentiated. The unsaturated carbon atoms in the alpha- and beta-acyl chains were also resolved into two signals except that of the C-11 olefinic carbon. Sandal (Santalum album) wood seed oil (a source of santalbic acid) was separated by silica chromatography into three fractions. The least polar fraction (7.2 wt%) contained TAG which had a random distribution of saturated and unsaturated fatty acids, of which oleic acid (69%) was the predominant component. The second fraction (3.8 wt%) contained santalbic acid (58%) and oleic acid (28%) together with some other normal fatty acids. Santalbic acid in this fraction was found in both the alpha- and beta-acyl positions of the glycerol "backbone." The most polar fraction (89 wt%) consisted of TAG containing santalbic acid only. The distribution of the various fatty acids on the glycerol "backbone" was supported by the results from the 13C NMR spectroscopic analysis.

  6. Electrophoretically mediated microanalysis of leucine aminopeptidase using two-photon excited fluorescence detection on a microchip.

    PubMed

    Zugel, S A; Burke, B J; Regnier, F E; Lytle, F E

    2000-11-15

    Two-photon excited fluorescence detection was performed on a microfabricated electrophoresis chip. A calibration curve of the fluorescent tag beta-naphthylamine was performed, resulting in a sensitivity of 2.5 x 10(9) counts M(-1) corresponding to a detection limit of 60 nM. Additionally, leucine aminopeptidase was assayed on the chip using electrophoretically mediated microanalysis. The differential electroosmotic mobilities of the enzyme and substrate, L-leucine beta-naphthylamide, allowed for efficient mixing in an open channel, resulting in the detection of a 30 nM enzyme solution under constant potential. A zero potential incubation for 1 min yielded a calculated detection limit of 4 nM enzyme.

  7. RAR Beta: Actions in Prostate Cancer

    DTIC Science & Technology

    2005-04-01

    spermine Nl-acetyltransferase Although prostate cancer can be clinically managed in its (SSAT) regulates the catabolism and export of intracellu- early...in anticancer strategies for spermidine and spermine pools were minimally de- some time (4). Various antagonists such as the ODC inhibitor creased due...Spm, spermine ; SSAT, spermidine/ spermine N’-acetyl- dressed: Dept. of Pharmacology and Therapeutics, Roswell Park Cancer transferase; Tag, SV40 large T

  8. Power Electronics Design of a Solar Powered In-car Wireless Tag for Asset Tracking and Parking Applications

    NASA Astrophysics Data System (ADS)

    Zhu, D.; Henaut, J.; Beeby, S. P.

    2014-11-01

    This paper reports the design and testing of a power conditioning circuit for a solar powered in-car wireless tag for asset tracking and parking application. Existing long range asset tracking is based on the GSM/GPRS network, which requires expensive subscriptions. The EU FP7 project CEWITT aims at developing a credit card sized autonomous wireless tag with GNSS geo-positioning capabilities to ensure the integrity and cost effectiveness for parking applications. It was found in previous research that solar cells are the most suitable energy sources for this application. This study focused on the power electronics design for the wireless tag. A suitable solar cell was chosen for its high power density. Charging circuit, hysteresis control circuit and LDO were designed and integrated to meet the system requirement. Test results showed that charging efficiency of 80 % had been achieved.

  9. Analysis and expression of the alpha-expansin and beta-expansin gene families in maize

    NASA Technical Reports Server (NTRS)

    Wu, Y.; Meeley, R. B.; Cosgrove, D. J.

    2001-01-01

    Expansins comprise a multigene family of proteins in maize (Zea mays). We isolated and characterized 13 different maize expansin cDNAs, five of which are alpha-expansins and eight of which are beta-expansins. This paper presents an analysis of these 13 expansins, as well as an expression analysis by northern blotting with materials from young and mature maize plants. Some expansins were expressed in restricted regions, such as the beta-expansins ExpB1 (specifically expressed in maize pollen) and ExpB4 (expressed principally in young husks). Other expansins such as alpha-expansin Exp1 and beta-expansin ExpB2 were expressed in several organs. The expression of yet a third group was not detected in the selected organs and tissues. An analysis of expansin sequences from the maize expressed sequence tag collection is also presented. Our results indicate that expansin genes may have general, overlapping expression in some instances, whereas in other cases the expression may be highly specific and limited to a single organ or cell type. In contrast to the situation in Arabidopsis, beta-expansins in maize seem to be more numerous and more highly expressed than are alpha-expansins. The results support the concept that beta-expansins multiplied and evolved special functions in the grasses.

  10. Cloning and characterization of ginsenoside Ra1-hydrolyzing beta-D-xylosidase from Bifidobacterium breve K-110.

    PubMed

    Hyun, Yang-Jin; Kim, Bomi; Kim, Dong-Hyun

    2012-04-01

    beta-D-Xylosidase (E.C. 3.2.1.37) from Bifidobacterium breve K-110, which hydrolyzes ginsenoside Ra1 to ginsenoside Rb2, was cloned and expressed in Escherichia coli. The (His6)-tagged recombinant enzyme, designated as XlyBK- 110, was efficiently purified using Ni²⁺-affinity chromatography (109.9-fold, 84% yield). The molecular mass of XylBK- 100 was found to be 55.7 kDa by SDS-PAGE. Its sequence revealed a 1,347 bp open reading frame (ORF) encoding a protein containing 448 amino acids, which showed 82% identity (DNA) to the previously reported glycosyl hydrolase family 30 of Bifidobacterium adolescentis ATCC 15703. The Km and Vmax values toward p-nitrophenyl-beta-D-xylopyranoside (pNPX) were 1.45mM and 10.75 micromol/min/mg, respectively. This enzyme had pH and temperature optima at 6.0 and 45 degrees C, respectively. XylBK-110 acted to the greatest extent on xyloglucosyl kakkalide, followed by pNPX and ginsenoside Ra1, but did not act on p-nitrophenyl-alpha-Larabinofuranoside, p-nitrophenyl-beta-D-glucopyranoside, or p-nitrophenyl-beta-D-fucopyranoside. In conclusion, this is the first report on the cloning and expression of beta-Dxylosidase- hydrolyzing ginsenoside Ra1 and kakkalide from human intestinal microflora.

  11. Exercise and Beta-Glucan Consumption (Saccharomyces cerevisiae) Improve the Metabolic Profile and Reduce the Atherogenic Index in Type 2 Diabetic Rats (HFD/STZ)

    PubMed Central

    Andrade, Eric Francelino; Lima, Andressa Ribeiro Veiga; Nunes, Ingrid Edwiges; Orlando, Débora Ribeiro; Gondim, Paula Novato; Zangeronimo, Márcio Gilberto; Alves, Fernando Henrique Ferrari; Pereira, Luciano José

    2016-01-01

    Physical activity and the ingestion of dietary fiber are non-drug alternatives commonly used as adjuvants to glycemic control in diabetic individuals. Among these fibers, we can highlight beta-glucans. However, few studies have compared isolated and synergic effects of physical exercise and beta-glucan ingestion, especially in type 2 diabetic rats. Therefore, we evaluated the effects beta-glucan (Saccharomyces cerevisiae) consumption, associated or not to exercise, on metabolic parameters of diabetic Wistar rats. The diabetes mellitus (DM) was induced by high-fat diet (HFD) associated with a low dose of streptozotocin (STZ—35 mg/kg). Trained groups were submitted to eight weeks of exercise in aquatic environment. In the last 28 days of experiment, animals received 30 mg/kg/day of beta-glucan by gavage. Isolated use of beta-glucan decreased glucose levels in fasting, Glycated hemoglobin (HbA1c), triglycerides (TAG), total cholesterol (TC), low-density lipoprotein (LDL-C), the atherogenic index of plasma. Exercise alone also decreased blood glucose levels, HbA1c, and renal lesions. An additive effect for reducing the atherogenic index of plasma and renal lesions was observed when both treatments were combined. It was concluded that both beta-glucan and exercise improved metabolic parameters in type 2 (HFD/STZ) diabetic rats. PMID:27999319

  12. Analysis of beta-carotene hydroxylase gene cDNA isolated from the American oil-palm (Elaeis oleifera) mesocarp tissue cDNA library

    PubMed Central

    Bhore, Subhash J; Kassim, Amelia; Loh, Chye Ying; Shah, Farida H

    2010-01-01

    It is well known that the nutritional quality of the American oil-palm (Elaeis oleifera) mesocarp oil is superior to that of African oil-palm (Elaeis guineensis Jacq. Tenera) mesocarp oil. Therefore, it is of important to identify the genetic features for its superior value. This could be achieved through the genome sequencing of the oil-palm. However, the genome sequence is not available in the public domain due to commercial secrecy. Hence, we constructed a cDNA library and generated expressed sequence tags (3,205) from the mesocarp tissue of the American oil-palm. We continued to annotate each of these cDNAs after submitting to GenBank/DDBJ/EMBL. A rough analysis turned our attention to the beta-carotene hydroxylase (Chyb) enzyme encoding cDNA. Then, we completed the full sequencing of cDNA clone for its both strands using M13 forward and reverse primers. The full nucleotide and protein sequence was further analyzed and annotated using various Bioinformatics tools. The analysis results showed the presence of fatty acid hydroxylase superfamily domain in the protein sequence. The multiple sequence alignment of selected Chyb amino acid sequences from other plant species and algal members with E. oleifera Chyb using ClustalW and its phylogenetic analysis suggest that Chyb from monocotyledonous plant species, Lilium hubrid, Crocus sativus and Zea mays are the most evolutionary related with E. oleifera Chyb. This study reports the annotation of E. oleifera Chyb. Abbreviations ESTs - expressed sequence tags, EoChyb - Elaeis oleifera beta-carotene hydroxylase, MC - main cluster PMID:21364789

  13. Ontologies and tag-statistics

    NASA Astrophysics Data System (ADS)

    Tibély, Gergely; Pollner, Péter; Vicsek, Tamás; Palla, Gergely

    2012-05-01

    Due to the increasing popularity of collaborative tagging systems, the research on tagged networks, hypergraphs, ontologies, folksonomies and other related concepts is becoming an important interdisciplinary area with great potential and relevance for practical applications. In most collaborative tagging systems the tagging by the users is completely ‘flat’, while in some cases they are allowed to define a shallow hierarchy for their own tags. However, usually no overall hierarchical organization of the tags is given, and one of the interesting challenges of this area is to provide an algorithm generating the ontology of the tags from the available data. In contrast, there are also other types of tagged networks available for research, where the tags are already organized into a directed acyclic graph (DAG), encapsulating the ‘is a sub-category of’ type of hierarchy between each other. In this paper, we study how this DAG affects the statistical distribution of tags on the nodes marked by the tags in various real networks. The motivation for this research was the fact that understanding the tagging based on a known hierarchy can help in revealing the hidden hierarchy of tags in collaborative tagging systems. We analyse the relation between the tag-frequency and the position of the tag in the DAG in two large sub-networks of the English Wikipedia and a protein-protein interaction network. We also study the tag co-occurrence statistics by introducing a two-dimensional (2D) tag-distance distribution preserving both the difference in the levels and the absolute distance in the DAG for the co-occurring pairs of tags. Our most interesting finding is that the local relevance of tags in the DAG (i.e. their rank or significance as characterized by, e.g., the length of the branches starting from them) is much more important than their global distance from the root. Furthermore, we also introduce a simple tagging model based on random walks on the DAG, capable of reproducing the main statistical features of tag co-occurrence. This model has high potential for further practical applications, e.g., it can provide the starting point for a benchmark system in ontology retrieval or it may help pinpoint unusual correlations in the co-occurrence of tags.

  14. Recombinant protein expression and purification: a comprehensive review of affinity tags and microbial applications.

    PubMed

    Young, Carissa L; Britton, Zachary T; Robinson, Anne S

    2012-05-01

    Protein fusion tags are indispensible tools used to improve recombinant protein expression yields, enable protein purification, and accelerate the characterization of protein structure and function. Solubility-enhancing tags, genetically engineered epitopes, and recombinant endoproteases have resulted in a versatile array of combinatorial elements that facilitate protein detection and purification in microbial hosts. In this comprehensive review, we evaluate the most frequently used solubility-enhancing and affinity tags. Furthermore, we provide summaries of well-characterized purification strategies that have been used to increase product yields and have widespread application in many areas of biotechnology including drug discovery, therapeutics, and pharmacology. This review serves as an excellent literature reference for those working on protein fusion tags. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. An RF-only ion-funnel for extraction from high-pressure gases

    DOE PAGES

    Brunner, T.; Fudenberg, D.; Varentsov, V.; ...

    2015-01-27

    An RF ion-funnel technique has been developed to extract ions from a high-pressure (10 bar) noble-gas environment into a vacuum (10 -6 mbar). Detailed simulations have been performed and a prototype has been developed for the purpose of extracting 136Ba ions from Xe gas with high efficiency. With this prototype, ions have been extracted for the first time from high-pressure xenon gas and argon gas. Systematic studies have been carried out and compared to simulations. This demonstration of extraction of ions, with mass comparable to that of the gas generating the high-pressure, has applications to Ba tagging from a Xe-gasmore » time-projection chamber for double-beta decay, as well as to the general problem of recovering trace amounts of an ionized element in a heavy (m > 40 u) carrier gas.« less

  16. IM-TORNADO: a tool for comparison of 16S reads from paired-end libraries.

    PubMed

    Jeraldo, Patricio; Kalari, Krishna; Chen, Xianfeng; Bhavsar, Jaysheel; Mangalam, Ashutosh; White, Bryan; Nelson, Heidi; Kocher, Jean-Pierre; Chia, Nicholas

    2014-01-01

    16S rDNA hypervariable tag sequencing has become the de facto method for accessing microbial diversity. Illumina paired-end sequencing, which produces two separate reads for each DNA fragment, has become the platform of choice for this application. However, when the two reads do not overlap, existing computational pipelines analyze data from read separately and underutilize the information contained in the paired-end reads. We created a workflow known as Illinois Mayo Taxon Organization from RNA Dataset Operations (IM-TORNADO) for processing non-overlapping reads while retaining maximal information content. Using synthetic mock datasets, we show that the use of both reads produced answers with greater correlation to those from full length 16S rDNA when looking at taxonomy, phylogeny, and beta-diversity. IM-TORNADO is freely available at http://sourceforge.net/projects/imtornado and produces BIOM format output for cross compatibility with other pipelines such as QIIME, mothur, and phyloseq.

  17. Reliable location-based services from radio navigation systems.

    PubMed

    Qiu, Di; Boneh, Dan; Lo, Sherman; Enge, Per

    2010-01-01

    Loran is a radio-based navigation system originally designed for naval applications. We show that Loran-C's high-power and high repeatable accuracy are fantastic for security applications. First, we show how to derive a precise location tag--with a sensitivity of about 20 meters--that is difficult to project to an exact location. A device can use our location tag to block or allow certain actions, without knowing its precise location. To ensure that our tag is reproducible we make use of fuzzy extractors, a mechanism originally designed for biometric authentication. We build a fuzzy extractor specifically designed for radio-type errors and give experimental evidence to show its effectiveness. Second, we show that our location tag is difficult to predict from a distance. For example, an observer cannot predict the location tag inside a guarded data center from a few hundreds of meters away. As an application, consider a location-aware disk drive that will only work inside the data center. An attacker who steals the device and is capable of spoofing Loran-C signals, still cannot make the device work since he does not know what location tag to spoof. We provide experimental data supporting our unpredictability claim.

  18. Social Tagging of Mission Data

    NASA Technical Reports Server (NTRS)

    Norris, Jeffrey S.; Wallick, Michael N.; Joswig, Joseph C.; Powell, Mark W.; Torres, Recaredo J.; Mittman, David S.; Abramyan, Lucy; Crockett, Thomas M.; Shams, Khawaja S.; Fox, Jason M.; hide

    2010-01-01

    Mars missions will generate a large amount of data in various forms, such as daily plans, images, and scientific information. Often, there is a semantic linkage between images that cannot be captured automatically. Software is needed that will provide a method for creating arbitrary tags for this mission data so that items with a similar tag can be related to each other. The tags should be visible and searchable for all users. A new routine was written to offer a new and more flexible search option over previous applications. This software allows users of the MSLICE program to apply any number of arbitrary tags to a piece of mission data through a MSLICE search interface. The application of tags creates relationships between data that did not previously exist. These tags can be easily removed and changed, and contain enough flexibility to be specifically configured for any mission. This gives users the ability to quickly recall or draw attention to particular pieces of mission data, for example: Give a semantic and meaningful description to mission data; for example, tag all images with a rock in them with the tag "rock." Rapidly recall specific and useful pieces of data; for example, tag a plan as"driving template." Call specific data to a user s attention; for example, tag a plan as "for:User." This software is part of the MSLICE release, which was written in Java. It will run on any current Windows, Macintosh, or Linux system.

  19. Integrated Management and Visualization of Electronic Tag Data with Tagbase

    PubMed Central

    Lam, Chi Hin; Tsontos, Vardis M.

    2011-01-01

    Electronic tags have been used widely for more than a decade in studies of diverse marine species. However, despite significant investment in tagging programs and hardware, data management aspects have received insufficient attention, leaving researchers without a comprehensive toolset to manage their data easily. The growing volume of these data holdings, the large diversity of tag types and data formats, and the general lack of data management resources are not only complicating integration and synthesis of electronic tagging data in support of resource management applications but potentially threatening the integrity and longer-term access to these valuable datasets. To address this critical gap, Tagbase has been developed as a well-rounded, yet accessible data management solution for electronic tagging applications. It is based on a unified relational model that accommodates a suite of manufacturer tag data formats in addition to deployment metadata and reprocessed geopositions. Tagbase includes an integrated set of tools for importing tag datasets into the system effortlessly, and provides reporting utilities to interactively view standard outputs in graphical and tabular form. Data from the system can also be easily exported or dynamically coupled to GIS and other analysis packages. Tagbase is scalable and has been ported to a range of database management systems to support the needs of the tagging community, from individual investigators to large scale tagging programs. Tagbase represents a mature initiative with users at several institutions involved in marine electronic tagging research. PMID:21750734

  20. Near-Field Chipless Radio-Frequency Identification (RFID) Sensing and Identification System with Switching Reading.

    PubMed

    Paredes, Ferran; Herrojo, Cristian; Mata-Contreras, Javier; Moras, Miquel; Núñez, Alba; Ramon, Eloi; Martín, Ferran

    2018-04-09

    A chipless radio-frequency identification (chipless-RFID) and sensing system, where tags are read by proximity (near-field) through a switch, is presented. The tags consist of a set of identical resonant elements (split-ring resonators or SRRs), printed or etched at predefined and equidistant positions, forming a linear chain, each SRR providing a bit of information. The logic state ('1' or '0') associated with each resonator depends on whether it is present or not in the predefined position. The reader is an array of power splitters used to feed a set of SRR-loaded transmission lines (in equal number to the number of resonant elements, or bits, of the tag). The feeding (interrogation) signal is a harmonic (single-tone) signal tuned to a frequency in the vicinity of the fundamental resonance of the SRRs. The set of SRR-loaded lines must be designed so that the corresponding SRRs are in perfect alignment with the SRRs of the tag, provided the tag is positioned on top of the reader. Thus, in a reading operation, as long as the tag is very close to the reader, the SRRs of the tag modify (decrease) the transmission coefficient of the corresponding reader line (through electromagnetic coupling between both SRRs), and the amplitude of the output signal is severely reduced. Therefore, the identification (ID) code of the tag is contained in the amplitudes of the output signals of the SRR-loaded lines, which can be inferred sequentially by means of a switching system. Unlike previous chipless-RFID systems based on near-field and sequential bit reading, the tags in the proposed system can be merely positioned on top of the reader, conveniently aligned, without the need to mechanically place them across the reader. Since tag reading is only possible if the tag is very close to the reader, this system can be also used as a proximity sensor with applications such as target identification. The proposed chipless-RFID and sensing approach is validated by reading a designed 4-bit tag. For identification purposes, this system is of special interest in applications where a low number of bits suffice, and tag reading by proximity is acceptable (or even convenient). Applications mostly related to secure paper, particularly involving a limited number of items (e.g., exams, ballots, etc.), in order to provide authenticity and avoid counterfeiting, are envisaged. As a proximity sensor, the system may be of use in detecting and distinguishing different targets in applications such as smart packaging.

  1. Near-Field Chipless Radio-Frequency Identification (RFID) Sensing and Identification System with Switching Reading

    PubMed Central

    Mata-Contreras, Javier; Moras, Miquel; Ramon, Eloi; Martín, Ferran

    2018-01-01

    A chipless radio-frequency identification (chipless-RFID) and sensing system, where tags are read by proximity (near-field) through a switch, is presented. The tags consist of a set of identical resonant elements (split-ring resonators or SRRs), printed or etched at predefined and equidistant positions, forming a linear chain, each SRR providing a bit of information. The logic state (‘1’ or ‘0’) associated with each resonator depends on whether it is present or not in the predefined position. The reader is an array of power splitters used to feed a set of SRR-loaded transmission lines (in equal number to the number of resonant elements, or bits, of the tag). The feeding (interrogation) signal is a harmonic (single-tone) signal tuned to a frequency in the vicinity of the fundamental resonance of the SRRs. The set of SRR-loaded lines must be designed so that the corresponding SRRs are in perfect alignment with the SRRs of the tag, provided the tag is positioned on top of the reader. Thus, in a reading operation, as long as the tag is very close to the reader, the SRRs of the tag modify (decrease) the transmission coefficient of the corresponding reader line (through electromagnetic coupling between both SRRs), and the amplitude of the output signal is severely reduced. Therefore, the identification (ID) code of the tag is contained in the amplitudes of the output signals of the SRR-loaded lines, which can be inferred sequentially by means of a switching system. Unlike previous chipless-RFID systems based on near-field and sequential bit reading, the tags in the proposed system can be merely positioned on top of the reader, conveniently aligned, without the need to mechanically place them across the reader. Since tag reading is only possible if the tag is very close to the reader, this system can be also used as a proximity sensor with applications such as target identification. The proposed chipless-RFID and sensing approach is validated by reading a designed 4-bit tag. For identification purposes, this system is of special interest in applications where a low number of bits suffice, and tag reading by proximity is acceptable (or even convenient). Applications mostly related to secure paper, particularly involving a limited number of items (e.g., exams, ballots, etc.), in order to provide authenticity and avoid counterfeiting, are envisaged. As a proximity sensor, the system may be of use in detecting and distinguishing different targets in applications such as smart packaging. PMID:29642560

  2. PPARgamma agonists inhibit TGF-beta-PKA signaling in glomerulosclerosis.

    PubMed

    Zou, Rong; Xu, Gang; Liu, Xiao-cheng; Han, Min; Jiang, Jing-jing; Huang, Qian; He, Yong; Yao, Ying

    2010-01-01

    To study the probable mechanisms of the anti-glomerulosclerosis effects induced by peroxisome proliferator-activated receptor gamma (PPARgamma) agonists in rat intraglomerular mesangial cells (MCs). Cells were transfected with the pTAL-PPRE-tk-Luc(+) plasmid and then treated with different concentrations of PPARgamma agonist, either troglitazone or telmisartan, for the indicated times. Promega luciferase assays were subsequently used for the detection of PPARgamma activation. Protein expression levels were assessed by Western blot, and PepTag assays were used for the non-radioactive detection of protein kinase A (PKA) activity. The deposition of alpha-smooth muscle actin (alpha-SMA) and p-cyclic AMP responsive element binding protein (pCREB) were analyzed by confocal laser scanning. Both troglitazone and telmisartan remarkably inhibit the PKA activation and pCREB expression that is stimulated by TGF-beta. The PPARgamma agonists also inhibited alpha-SMA and collagen IV protein expression by blocking PKA activation. PPARgamma ligands effectively suppress the activation of MCs and the accumulation of collagen IV stimulated by TGF-beta in vitro. The renal protection provided by PPARgamma agonists is partly mediated via their blockade of TGF-beta/PKA signaling.

  3. PNNL Fish Telemetry: Improving the Sustainability of Hydropower One Tag at a Time

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    None

    Evaluating the environmental impact of hydropower is critical to the growth, development, and maintenance of this vital energy source. Operators, developers, and regulators often turn to animal telemetry to measure the effect of dams on fish populations. PNNL’s tagging portfolio consists of a range of tags suitable for a variety of applications from active tags slightly larger than a grain of rice to larger tags that power themselves via a fish’s natural movement.

  4. Application of restoration scenarios to basin-scale demographics of coho salmon inferred from pit-tags

    EPA Science Inventory

    Effective habitat restoration planning requires correctly anticipating demographic responses to altered habitats. New applications of Passive Integrated Transponder (PIT) tag technology to fish-habitat research have provided critical insights into fish movement, growth, and surv...

  5. Retention of coded wire tags, and their effect on maturation and survival of yellow mealworms (Coleoptera: Tenebrionidae)

    USGS Publications Warehouse

    Schaffler, James J.; Isely, J.J.

    2001-01-01

    This study demonstrates that coded wire tags can be used to mark certain insect larvae without adverse effects on maturation, and that tags are retained through the adult phase in high enough proportion for practical application. Coded wire tags also offer the benefit that marked organisms can be identified to the batch or individual level.

  6. Achievable Performance and Effective Interrogator Design for SAW RFID Sensor Tags

    NASA Technical Reports Server (NTRS)

    Barton Richard J.

    2012-01-01

    For many NASA missions, remote sensing is a critical application that supports activities such as environmental monitoring, planetary science, structural shape and health monitoring, non-destructive evaluation, etc. The utility of the remote sensing devices themselves is greatly increased if they are passive V that is, they do not require any on-board power supply such as batteries V and if they can be identified uniquely during the sensor interrogation process. Additional passive sensor characteristics that enable greater utilization in space applications are small size and weight, long read ranges with low interrogator power, ruggedness, and operability in extreme environments (vacuum, extreme high/low temperature, high radiation, etc.) In this paper, we consider one very promising passive sensor technology, called surface acoustic wave (SAW) radio-frequency identification (RFID), that satisfies all of these criteria. In general, RFID is a method of identifying items using radio waves to interrogate tags encoded with a unique identifier that are affixed to the items of interest. In the case of passive tags, only the interrogator, which transmits power to the tags in the form of radio-frequency electromagnetic radiation, requires access to a power supply. Passive RFID technologies are used today in many applications, including asset tracking and management, security and access control, and remote sensing. To date, most of the development and application in RFID technology has focused on either asset/inventory tracking and control or security and access control because these are the largest commercial application areas. Recently however, there has been growing interest in using passive RFID technology for remote sensing applications, and SAW devices are at the forefront of RFID sensing technology development. Although SAW RFID tags have great potential for use in numerous space-based remote sensing applications, the limited collision resolution capability of current generation tags limits the performance in a cluttered sensing environment. That is, as more SAW-based sensors are added to the environment, numerous tag responses are superimposed at the receiver and decoding all or even a subset of the telemetry becomes increasingly difficult. Background clutter generated by reflectors other than the sensors themselves is also a problem, as is multipath interference and signal distortion, but the limiting factor in many remote sensing applications can be expected to be tag mutual interference. This problem may be greatly mitigated by proper design of the SAW tag waveform, but that remains an open research problem, and in the meantime, several other related questions remain to be answered including: (1) What are the fundamental relationships between tag parameters such as bit-rate, time-bandwidth-product, SNR, and achievable collision resolution? (2) What are the differences in optimal or near-optimal interrogator designs between noise-limited environments and interference-limited environments? (3) What are the performance characteristics of different interrogator designs in term of parameters such as transmitter power level, range, and number of interfering tags? In this paper, we will present the results of a research effort aimed at providing at least partial answers to all of these questions.

  7. Bioconjugates of luminescent CdSe-ZnS quantum dots with an engineered two-domain protein G for use in fluoroimmunoassays

    NASA Astrophysics Data System (ADS)

    Tran, Phan T.; Goldman, Ellen R.; Mattoussi, Hedi M.; Anderson, George P.; Mauro, J. Matthew

    2001-06-01

    Colloidal semiconductor quantum dots (QDs) seem suitable for labeling certain biomolecules for use in fluorescent tagging applications, such as fluoro-immunoassays. Compared to organic dye labels, Qds are resistant to photo-degradation, and these luminescent nanoparticles have size-dependent emission spectra spanning a wide range of wavelengths in the visible and near IR. We previously described an electrostatic self-assembly approach for conjugating highly luminescent colloidal CdSe-ZnS core-shell Qds with engineered two-domain recombinant proteins. Here we describe the application of this approach to prepare QD conjugates with the (Beta) 2 immunoglobin G (IgG) binding domain of streptococcal protein G (PG) appended with a basic lucine zipper attachment domain (PG-zb). We also demonstrate that the QD/PG conjugates retain their ability to bind IgG antibodies, and that a specific antibody coupled to QD via the PG functional domain efficiently binds its antigen. These preliminary results indicate that electrostatically self-assembled QD/PG-zb/IgG bioconjugates can be used in fluoro-immunoassays.

  8. Postprandial lipid responses to an alpha-linolenic acid-rich oil, olive oil and butter in women: a randomized crossover trial.

    PubMed

    Svensson, Julia; Rosenquist, Anna; Ohlsson, Lena

    2011-06-28

    Postprandial lipaemia varies with gender and the composition of dietary fat due to the partitioning of fatty acids between beta-oxidation and incorporation into triacylglycerols (TAGs). Increasing evidence highlights the importance of postprandial measurements to evaluate atherogenic risk. Postprandial effects of alpha-linolenic acid (ALA) in women are poorly characterized. We therefore studied the postprandial lipid response of women to an ALA-rich oil in comparison with olive oil and butter, and characterized the fatty acid composition of total lipids, TAGs, and non-esterified fatty acids (NEFAs) in plasma. A randomized crossover design (n = 19) was used to compare the postprandial effects of 3 meals containing 35 g fat. Blood samples were collected at regular intervals for 7 h. Statistical analysis was carried out with ANOVA (significant difference = P < 0.05). No significant difference was seen in incremental area under the curve (iAUC) plasma-TAG between the meals. ALA and oleic acid levels were significantly increased in plasma after ALA-rich oil and olive oil meals, respectively. Palmitic acid was significantly increased in plasma-TAG after the butter meal. The ratios of 18:2 n-6 to18:3 n-3 in plasma-TAGs, three and seven hours after the ALA-rich oil meal, were 1.5 and 2.4, respectively. The corresponding values after the olive oil meal were: 13.8 and 16.9; and after the butter meal: 9.0 and 11.6. The postprandial p-TAG and NEFA response in healthy pre-menopausal women was not significantly different after the intake of an ALA-rich oil, olive oil and butter. The ALA-rich oil significantly affected different plasma lipid fractions and improved the ratio of n-6 to n-3 fatty acids several hours postprandially.

  9. Calibration of LiBaF3: Ce Scintillator for Fission Spectrum Neutrons

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Reeder, Paul L.; Bowyer, Sonya M.

    2002-05-21

    The scintillator LiBaF3 doped with small amounts of Ce+3 has the ability to distinguish heavy charged particles (p, d, t, or a) from beta and/or gamma radiation based on the presence or absence of ns components in the scintillation light output. Because the neutron capture reaction on 6Li produces recoil alphas and tritons, this scintillator also discriminates between neutron induced events and beta or gamma interactions. An experimental technique using a time-tagged 252Cf source has been used to measure the efficiency of this scintillator for neutron capture, the calibration of neutron capture pulse height, and the pulse height resolution -more » all as a function of incident neutron energy.« less

  10. Does coffee consumption impact on heaviness of smoking?

    PubMed Central

    Ware, Jennifer J.; Tanner, Julie‐Anne; Taylor, Amy E.; Bin, Zhao; Haycock, Philip; Bowden, Jack; Rogers, Peter J.; Davey Smith, George; Tyndale, Rachel F.

    2017-01-01

    Abstract Background and Aims Coffee consumption and cigarette smoking are strongly associated, but whether this association is causal remains unclear. We sought to: (1) determine whether coffee consumption influences cigarette smoking causally, (2) estimate the magnitude of any association and (3) explore potential mechanisms. Design We used Mendelian randomization (MR) analyses of observational data, using publicly available summarized data from the Tobacco and Genetics (TAG) consortium, individual‐level data from the UK Biobank and in‐vitro experiments of candidate compounds. Setting The TAG consortium includes data from studies in several countries. The UK Biobank includes data from men and women recruited across England, Wales and Scotland. Participants The TAG consortium provided data on n ≤ 38 181 participants. The UK Biobank provided data on 8072 participants. Measurements In MR analyses, the exposure was coffee consumption (cups/day) and the outcome was heaviness of smoking (cigarettes/day). In our in‐vitro experiments we assessed the effect of caffeic acid, quercetin and p‐coumaric acid on the rate of nicotine metabolism in human liver microsomes and cDNA‐expressed human CYP2A6. Findings Two‐sample MR analyses of TAG consortium data indicated that heavier coffee consumption might lead to reduced heaviness of smoking [beta = −1.49, 95% confidence interval (CI) = −2.88 to −0.09]. However, in‐vitro experiments found that the compounds investigated are unlikely to inhibit significantly the rate of nicotine metabolism following coffee consumption. Further MR analyses in UK Biobank found no evidence of a causal relationship between coffee consumption and heaviness of smoking (beta = 0.20, 95% CI = –1.72 to 2.12). Conclusions Amount of coffee consumption is unlikely to have a major causal impact upon amount of cigarette smoking. If it does influence smoking, this is not likely to operate via effects of caffeic acid, quercetin or p‐coumaric acid on nicotine metabolism. The observational association between coffee consumption and cigarette smoking may be due to smoking impacting on coffee consumption or confounding. PMID:28556459

  11. Does coffee consumption impact on heaviness of smoking?

    PubMed

    Ware, Jennifer J; Tanner, Julie-Anne; Taylor, Amy E; Bin, Zhao; Haycock, Philip; Bowden, Jack; Rogers, Peter J; Davey Smith, George; Tyndale, Rachel F; Munafò, Marcus R

    2017-10-01

    Coffee consumption and cigarette smoking are strongly associated, but whether this association is causal remains unclear. We sought to: (1) determine whether coffee consumption influences cigarette smoking causally, (2) estimate the magnitude of any association and (3) explore potential mechanisms. We used Mendelian randomization (MR) analyses of observational data, using publicly available summarized data from the Tobacco and Genetics (TAG) consortium, individual-level data from the UK Biobank and in-vitro experiments of candidate compounds. The TAG consortium includes data from studies in several countries. The UK Biobank includes data from men and women recruited across England, Wales and Scotland. The TAG consortium provided data on n ≤ 38 181 participants. The UK Biobank provided data on 8072 participants. In MR analyses, the exposure was coffee consumption (cups/day) and the outcome was heaviness of smoking (cigarettes/day). In our in-vitro experiments we assessed the effect of caffeic acid, quercetin and p-coumaric acid on the rate of nicotine metabolism in human liver microsomes and cDNA-expressed human CYP2A6. Two-sample MR analyses of TAG consortium data indicated that heavier coffee consumption might lead to reduced heaviness of smoking [beta = -1.49, 95% confidence interval (CI) = -2.88 to -0.09]. However, in-vitro experiments found that the compounds investigated are unlikely to inhibit significantly the rate of nicotine metabolism following coffee consumption. Further MR analyses in UK Biobank found no evidence of a causal relationship between coffee consumption and heaviness of smoking (beta = 0.20, 95% CI = -1.72 to 2.12). Amount of coffee consumption is unlikely to have a major causal impact upon amount of cigarette smoking. If it does influence smoking, this is not likely to operate via effects of caffeic acid, quercetin or p-coumaric acid on nicotine metabolism. The observational association between coffee consumption and cigarette smoking may be due to smoking impacting on coffee consumption or confounding. © 2017 The Authors. Addiction published by John Wiley & Sons Ltd on behalf of Society for the Study of Addiction.

  12. Amyloid-beta peptide oligomers disrupt axonal transport through an NMDA receptor-dependent mechanism that is mediated by glycogen synthase kinase 3beta in primary cultured hippocampal neurons.

    PubMed

    Decker, Helena; Lo, Karen Y; Unger, Sandra M; Ferreira, Sergio T; Silverman, Michael A

    2010-07-07

    Disruption of axonal transport is a hallmark of several neurodegenerative diseases, including Alzheimer's disease (AD). Even though defective transport is considered an early pathologic event, the mechanisms by which neurodegenerative insults impact transport are poorly understood. We show that soluble oligomers of the amyloid-beta peptide (AbetaOs), increasingly recognized as the proximal neurotoxins in AD pathology, induce disruption of organelle transport in primary hippocampal neurons in culture. Live imaging of fluorescent protein-tagged organelles revealed a marked decrease in axonal trafficking of dense-core vesicles and mitochondria in the presence of 0.5 microm AbetaOs. NMDA receptor (NMDAR) antagonists, including d-AP5, MK-801, and memantine, prevented the disruption of trafficking, thereby identifying signals for AbetaO action at the cell membrane. Significantly, both pharmacological inhibition of glycogen synthase kinase-3beta (GSK-3beta) and transfection of neurons with a kinase-dead form of GSK-3beta prevented the transport defect. Finally, we demonstrate by biochemical and immunocytochemical means that AbetaOs do not affect microtubule stability, indicating that disruption of transport involves a more subtle mechanism than microtubule destabilization, likely the dysregulation of intracellular signaling cascades. Results demonstrate that AbetaOs negatively impact axonal transport by a mechanism that is initiated by NMDARs and mediated by GSK-3beta and establish a new connection between toxic Abeta oligomers and AD pathology.

  13. Envisioning the Future of Aquatic Animal Tracking: Technology, Science, and Application

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lennox, Robert J.; Aarestrup, Kim; Cooke, Steven J.

    Electronic tags have proven to be extremely useful for broadening our understanding of aquatic animals by answering diverse questions about their behaviours, physiologies, and life histories fundamental to ecology. Simultaneously, many applied conservation and management efforts are informed by animals tagged with electronic tags. In spite of the many advances in tracking software and hardware, an uncertain future in the world’s aquatic ecosystems portends great challenges for science. Aquatic animal tracking with electronic tags represents both the present and future of integrative biology and ecology in aquatic ecosystems. Here we identify what we regard as the future of aquatic animalmore » tracking in a horizon scanning exercise. We submit that the future of aquatic animal tracking will include opportunities for multi-platform tracking systems for simultaneously monitoring position, activity, physiology, and microhabitat of animals, improved data collection and accessibility with new infrastructure (e.g. tags, receivers) and cyberinfrastructure, and integrated tagging information with animal traits derived from biopsy during tagging. We discuss parallel needs and opportunities in areas related to the application of animal tracking in the future such as knowledge mobilization and governance.« less

  14. Analysis of aromatic catabolic pathways in Pseudomonas putida KT 2440 using a combined proteomic approach: 2-DE/MS and cleavable isotope-coded affinity tag analysis.

    PubMed

    Kim, Young Hwan; Cho, Kun; Yun, Sung-Ho; Kim, Jin Young; Kwon, Kyung-Hoon; Yoo, Jong Shin; Kim, Seung Il

    2006-02-01

    Proteomic analysis of Pseudomonas putida KT2440 cultured in monocyclic aromatic compounds was performed using 2-DE/MS and cleavable isotope-coded affinity tag (ICAT) to determine whether proteins involved in aromatic compound degradation pathways were altered as predicted by genomic analysis (Jiménez et al., Environ Microbiol. 2002, 4, 824-841). Eighty unique proteins were identified by 2-DE/MS or MS/MS analysis from P. putida KT2440 cultured in the presence of six different organic compounds. Benzoate dioxygenase (BenA, BenD) and catechol 1,2-dioxygenase (CatA) were induced by benzoate. Protocatechuate 3,4-dixoygenase (PcaGH) was induced by p-hydroxybenzoate and vanilline. beta-Ketoadipyl CoA thiolase (PcaF) and 3-oxoadipate enol-lactone hydrolase (PcaD) were induced by benzoate, p-hydroxybenzoate and vanilline, suggesting that benzoate, p-hydroxybenzoate and vanilline were degraded by different dioxygenases and then converged in the same beta-ketoadipate degradation pathway. An additional 110 proteins, including 19 proteins from 2-DE analysis, were identified by cleavable ICAT analysis for benzoate-induced proteomes, which complemented the 2-DE results. Phenylethylamine exposure induced beta-ketoacyl CoA thiolase (PhaD) and ring-opening enzyme (PhaL), both enzymes of the phenylacetate (pha) biodegradation pathway. Phenylalanine induced 4-hydroxyphenyl-pyruvate dioxygenase (Hpd) and homogentisate 1,2-dioxygenase (HmgA), key enzymes in the homogentisate degradation pathway. Alkyl hydroperoxide reductase (AphC) was induced under all aromatic compounds conditions. These results suggest that proteome analysis complements and supports predictive information obtained by genomic sequence analysis.

  15. A statistical method for assessing peptide identification confidence in accurate mass and time tag proteomics

    PubMed Central

    Stanley, Jeffrey R.; Adkins, Joshua N.; Slysz, Gordon W.; Monroe, Matthew E.; Purvine, Samuel O.; Karpievitch, Yuliya V.; Anderson, Gordon A.; Smith, Richard D.; Dabney, Alan R.

    2011-01-01

    Current algorithms for quantifying peptide identification confidence in the accurate mass and time (AMT) tag approach assume that the AMT tags themselves have been correctly identified. However, there is uncertainty in the identification of AMT tags, as this is based on matching LC-MS/MS fragmentation spectra to peptide sequences. In this paper, we incorporate confidence measures for the AMT tag identifications into the calculation of probabilities for correct matches to an AMT tag database, resulting in a more accurate overall measure of identification confidence for the AMT tag approach. The method is referred to as Statistical Tools for AMT tag Confidence (STAC). STAC additionally provides a Uniqueness Probability (UP) to help distinguish between multiple matches to an AMT tag and a method to calculate an overall false discovery rate (FDR). STAC is freely available for download as both a command line and a Windows graphical application. PMID:21692516

  16. Anchor and visible implant elastomer tag retention by hatchery rainbow trout stocked into an Ozark stream

    USGS Publications Warehouse

    Walsh, M.G.; Winkelman, D.L.

    2004-01-01

    As part of a study to evaluate the stocking of rainbow trout Oncorhynchus mykiss in an Oklahoma Ozark stream, we tagged 2,542 hatchery-reared rainbow trout (123-366 mm total length) with individually numbered Floy FD-68B anchor tags and visible implant fluorescent elastomer (VIE) tags. We experimentally stocked double-marked rainbow trout into a small northeastern Oklahoma stream from November 2001 to March 2002 and resampled them monthly from December 2001 to October 2002 by electrofishing. Anchor tag retention was 91% through 6 months, and VIE tag retention was 96% through 6 months despite extensive handling of fish within 24 h of tagging. Based on the ease of application, high visibility, and high retention observed in this study, we recommend the use of VIE tags as a batch mark in similarly sized, similarly pigmented fish. The retention of VIE tags was slightly higher than that of anchor tags, and cost per fish was less for VIE than for anchor tags. However, VIE tags would have limited utility if numerous individual tags are necessary; therefore, we recommend anchor tags as individual marks in similarly sized salmonids. Retention for both tag types was relatively high and could be corrected for when estimating population parameters from tagging data.

  17. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sorensen, Christina M.; Ding, Jie; Zhang, Qibin

    Objectives: To characterize the lipid profile of individuals with newly diagnosed type 1 diabetes mellitus using LC-MS-based lipidomics and the accurate mass and time (AMT) tag approach. Design and methods: Lipids were extracted from plasma and sera of 10 subjects from the Diabetes Antibody Standardization Program (years 2000-2005) and 10 non-diabetic subjects and analyzed by capillary liquid chromatography coupled with a hybrid ion-trap-Fourier transform ion cyclotron resonance mass spectrometer. Lipids were identified and quantified using the AMT tag approach. Results: Five hundred sixty lipid features differentiated (q < 0.05) diabetic from healthy individuals in a partial least-squares analysis, characterizing ofmore » individuals with recently diagnosed type 1 diabetes mellitus. Conclusions: A lipid profile associated with newly diagnosed type 1 diabetes may aid in further characterization of biochemical pathways involved in lipid regulation or mobilization and lipotoxicity of pancreatic beta-cells.« less

  18. Review on SAW RFID tags.

    PubMed

    Plessky, Victor P; Reindl, Leonhard M

    2010-03-01

    SAW tags were invented more than 30 years ago, but only today are the conditions united for mass application of this technology. The devices in the 2.4-GHz ISM band can be routinely produced with optical lithography, high-resolution radar systems can be built up using highly sophisticated, but low-cost RF-chips, and the Internet is available for global access to the tag databases. The "Internet of Things," or I-o-T, will demand trillions of cheap tags and sensors. The SAW tags can overcome semiconductor-based analogs in many aspects: they can be read at a distance of a few meters with readers radiating power levels 2 to 3 orders lower, they are cheap, and they can operate in robust environments. Passive SAW tags are easily combined with sensors. Even the "anti-collision" problem (i.e., the simultaneous reading of many nearby tags) has adequate solutions for many practical applications. In this paper, we discuss the state-of-the-art in the development of SAW tags. The design approaches will be reviewed and optimal tag designs, as well as encoding methods, will be demonstrated. We discuss ways to reduce the size and cost of these devices. A few practical examples of tags using a time-position coding with 10(6) different codes will be demonstrated. Phase-coded devices can additionally increase the number of codes at the expense of a reduction of reading distance. We also discuss new and exciting perspectives of using ultra wide band (UWB) technology for SAW-tag systems. The wide frequency band available for this standard provides a great opportunity for SAW tags to be radically reduced in size to about 1 x 1 mm(2) while keeping a practically infinite number of possible different codes. Finally, the reader technology will be discussed, as well as detailed comparison made between SAW tags and IC-based semiconductor device.

  19. Quenched substrates for live-cell labeling of SNAP-tagged fusion proteins with improved fluorescent background.

    PubMed

    Stöhr, Katharina; Siegberg, Daniel; Ehrhard, Tanja; Lymperopoulos, Konstantinos; Öz, Simin; Schulmeister, Sonja; Pfeifer, Andrea C; Bachmann, Julie; Klingmüller, Ursula; Sourjik, Victor; Herten, Dirk-Peter

    2010-10-01

    Recent developments in fluorescence microscopy raise the demands for bright and photostable fluorescent tags for specific and background free labeling in living cells. Aside from fluorescent proteins and other tagging methods, labeling of SNAP-tagged proteins has become available thereby increasing the pool of potentially applicable fluorescent dyes for specific labeling of proteins. Here, we report on novel conjugates of benzylguanine (BG) which are quenched in their fluorescence and become highly fluorescent upon labeling of the SNAP-tag, the commercial variant of the human O(6)-alkylguanosyltransferase (hAGT). We identified four conjugates showing a strong increase, i.e., >10-fold, in fluorescence intensity upon labeling of SNAP-tag in vitro. Moreover, we screened a subset of nine BG-dye conjugates in living Escherichia coli and found them all suited for labeling of the SNAP-tag. Here, quenched BG-dye conjugates yield a higher specificity due to reduced contribution from excess conjugate to the fluorescence signal. We further extended the application of these conjugates by labeling a SNAP-tag fusion of the Tar chemoreceptor in live E. coli cells and the eukaryotic transcription factor STAT5b in NIH 3T3 mouse fibroblast cells. Aside from the labeling efficiency and specificity in living cells, we discuss possible mechanisms that might be responsible for the changes in fluorescence emission upon labeling of the SNAP-tag, as well as problems we encountered with nonspecific labeling with certain conjugates in eukaryotic cells.

  20. Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging

    PubMed Central

    Provost, Christopher R.; Sun, Luo

    2010-01-01

    SNAP-tag and CLIP-tag protein labeling systems enable the specific, covalent attachment of molecules, including fluorescent dyes, to a protein of interest in live cells. These systems offer a broad selection of fluorescent substrates optimized for a range of imaging instrumentation. Once cloned and expressed, the tagged protein can be used with a variety of substrates for numerous downstream applications without having to clone again. There are two steps to using this system: cloning and expression of the protein of interest as a SNAP-tag fusion, and labeling of the fusion with the SNAP-tag substrate of choice. The SNAP-tag is a small protein based on human O6-alkylguanine-DNA-alkyltransferase (hAGT), a DNA repair protein. SNAP-tag labels are dyes conjugated to guanine or chloropyrimidine leaving groups via a benzyl linker. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag. CLIP-tag is a modified version of SNAP-tag, engineered to react with benzylcytosine rather than benzylguanine derivatives. When used in conjunction with SNAP-tag, CLIP-tag enables the orthogonal and complementary labeling of two proteins simultaneously in the same cells. PMID:20485262

  1. Energy-balanced algorithm for RFID estimation

    NASA Astrophysics Data System (ADS)

    Zhao, Jumin; Wang, Fangyuan; Li, Dengao; Yan, Lijuan

    2016-10-01

    RFID has been widely used in various commercial applications, ranging from inventory control, supply chain management to object tracking. It is necessary for us to estimate the number of RFID tags deployed in a large area periodically and automatically. Most of the prior works use passive tags to estimate and focus on designing time-efficient algorithms that can estimate tens of thousands of tags in seconds. But for a RFID reader to access tags in a large area, active tags are likely to be used due to their longer operational ranges. But these tags use their own battery as energy supplier. Hence, conserving energy for active tags becomes critical. Some prior works have studied how to reduce energy expenditure of a RFID reader when it reads tags IDs. In this paper, we study how to reduce the amount of energy consumed by active tags during the process of estimating the number of tags in a system and make the energy every tag consumed balanced approximately. We design energy-balanced estimation algorithm that can achieve our goal we mentioned above.

  2. Metal Stable Isotope Tagging: Renaissance of Radioimmunoassay for Multiplex and Absolute Quantification of Biomolecules.

    PubMed

    Liu, Rui; Zhang, Shixi; Wei, Chao; Xing, Zhi; Zhang, Sichun; Zhang, Xinrong

    2016-05-17

    The unambiguous quantification of biomolecules is of great significance in fundamental biological research as well as practical clinical diagnosis. Due to the lack of a detectable moiety, the direct and highly sensitive quantification of biomolecules is often a "mission impossible". Consequently, tagging strategies to introduce detectable moieties for labeling target biomolecules were invented, which had a long and significant impact on studies of biomolecules in the past decades. For instance, immunoassays have been developed with radioisotope tagging by Yalow and Berson in the late 1950s. The later languishment of this technology can be almost exclusively ascribed to the use of radioactive isotopes, which led to the development of nonradioactive tagging strategy-based assays such as enzyme-linked immunosorbent assay, fluorescent immunoassay, and chemiluminescent and electrochemiluminescent immunoassay. Despite great success, these strategies suffered from drawbacks such as limited spectral window capacity for multiplex detection and inability to provide absolute quantification of biomolecules. After recalling the sequences of tagging strategies, an apparent question is why not use stable isotopes from the start? A reasonable explanation is the lack of reliable means for accurate and precise quantification of stable isotopes at that time. The situation has changed greatly at present, since several atomic mass spectrometric measures for metal stable isotopes have been developed. Among the newly developed techniques, inductively coupled plasma mass spectrometry is an ideal technique to determine metal stable isotope-tagged biomolecules, for its high sensitivity, wide dynamic linear range, and more importantly multiplex and absolute quantification ability. Since the first published report by our group, metal stable isotope tagging has become a revolutionary technique and gained great success in biomolecule quantification. An exciting research highlight in this area is the development and application of the mass cytometer, which fully exploited the multiplexing potential of metal stable isotope tagging. It realized the simultaneous detection of dozens of parameters in single cells, accurate immunophenotyping in cell populations, through modeling of intracellular signaling network and undoubted discrimination of function and connection of cell subsets. Metal stable isotope tagging has great potential applications in hematopoiesis, immunology, stem cells, cancer, and drug screening related research and opened a post-fluorescence era of cytometry. Herein, we review the development of biomolecule quantification using metal stable isotope tagging. Particularly, the power of multiplex and absolute quantification is demonstrated. We address the advantages, applicable situations, and limitations of metal stable isotope tagging strategies and propose suggestions for future developments. The transfer of enzymatic or fluorescent tagging to metal stable isotope tagging may occur in many aspects of biological and clinical practices in the near future, just as the revolution from radioactive isotope tagging to fluorescent tagging happened in the past.

  3. Utility of biological sensor tags in animal conservation.

    PubMed

    Wilson, A D M; Wikelski, M; Wilson, R P; Cooke, S J

    2015-08-01

    Electronic tags (both biotelemetry and biologging platforms) have informed conservation and resource management policy and practice by providing vital information on the spatial ecology of animals and their environments. However, the extent of the contribution of biological sensors (within electronic tags) that measure an animal's state (e.g., heart rate, body temperature, and details of locomotion and energetics) is less clear. A literature review revealed that, despite a growing number of commercially available state sensor tags and enormous application potential for such devices in animal biology, there are relatively few examples of their application to conservation. Existing applications fell under 4 main themes: quantifying disturbance (e.g., ecotourism, vehicular and aircraft traffic), examining the effects of environmental change (e.g., climate change), understanding the consequences of habitat use and selection, and estimating energy expenditure. We also identified several other ways in which sensor tags could benefit conservation, such as determining the potential efficacy of management interventions. With increasing sensor diversity of commercially available platforms, less invasive attachment techniques, smaller device sizes, and more researchers embracing such technology, we suggest that biological sensor tags be considered a part of the necessary toolbox for conservation. This approach can measure (in real time) the state of free-ranging animals and thus provide managers with objective, timely, relevant, and accurate data to inform policy and decision making. © 2015 Society for Conservation Biology.

  4. Reliable Location-Based Services from Radio Navigation Systems

    PubMed Central

    Qiu, Di; Boneh, Dan; Lo, Sherman; Enge, Per

    2010-01-01

    Loran is a radio-based navigation system originally designed for naval applications. We show that Loran-C’s high-power and high repeatable accuracy are fantastic for security applications. First, we show how to derive a precise location tag—with a sensitivity of about 20 meters—that is difficult to project to an exact location. A device can use our location tag to block or allow certain actions, without knowing its precise location. To ensure that our tag is reproducible we make use of fuzzy extractors, a mechanism originally designed for biometric authentication. We build a fuzzy extractor specifically designed for radio-type errors and give experimental evidence to show its effectiveness. Second, we show that our location tag is difficult to predict from a distance. For example, an observer cannot predict the location tag inside a guarded data center from a few hundreds of meters away. As an application, consider a location-aware disk drive that will only work inside the data center. An attacker who steals the device and is capable of spoofing Loran-C signals, still cannot make the device work since he does not know what location tag to spoof. We provide experimental data supporting our unpredictability claim. PMID:22163532

  5. Comparing the hierarchy of author given tags and repository given tags in a large document archive

    NASA Astrophysics Data System (ADS)

    Tibély, Gergely; Pollner, Péter; Palla, Gergely

    2016-10-01

    Folksonomies - large databases arising from collaborative tagging of items by independent users - are becoming an increasingly important way of categorizing information. In these systems users can tag items with free words, resulting in a tripartite item-tag-user network. Although there are no prescribed relations between tags, the way users think about the different categories presumably has some built in hierarchy, in which more special concepts are descendants of some more general categories. Several applications would benefit from the knowledge of this hierarchy. Here we apply a recent method to check the differences and similarities of hierarchies resulting from tags given by independent individuals and from tags given by a centrally managed repository system. The results from our method showed substantial differences between the lower part of the hierarchies, and in contrast, a relatively high similarity at the top of the hierarchies.

  6. Design considerations and tradeoffs for passive RFID tags

    NASA Astrophysics Data System (ADS)

    Hussien, Faisal A.; Turker, Didem Z.; Srinivasan, Rangakrishnan; Mobarak, Mohamed S.; Cortes, Fernando P.; Sanchez-Sinencio, Edgar

    2005-06-01

    Radio Frequency Identification (RFID) systems are widely used in a variety of tracking, security and tagging applications. Their operation in non line-of-sight environments makes them superior over similar devices such as barcode and infrared tags. RFID systems span a wide range of applications: medical history storage, dental prosthesis tracking, oil drilling pipe and concrete stress monitoring, toll ways services, animal tracking applications, etc. Passive RFID tags generate their power from the incoming signal; therefore, they do not require a power source. Accordingly, minimizing the power consumption and the implementation area are usually the main design considerations. This paper presents a complete analysis on designing a passive RFID tag. A system design methodology is introduced including the main issues and tradeoffs between different design parameters. The uplink modulation techniques used (ASK, PSK, FSK, and PWM) are illustrated showing how to choose the appropriate signaling scheme for a specific data rate, a certain distance of operation and a limited power consumption budget. An antenna system (transmitter and receiver) is proposed providing the maximum distance of operation with the transmitted power stated by FCC regulations. The backscatter modulation scheme used in the downlink is shown whether to be ASK-BM or PSK-BM and the differences between them are discussed. The key building blocks such as the charge pump, voltage reference, and the regulator used to generate the DC supply voltage from the incoming RF signal are discussed along with their design tradeoffs. A complete architecture for a passive RFID tag is provided as an example to illustrate the proposed RFID tag design methodology.

  7. Social tagging in the life sciences: characterizing a new metadata resource for bioinformatics.

    PubMed

    Good, Benjamin M; Tennis, Joseph T; Wilkinson, Mark D

    2009-09-25

    Academic social tagging systems, such as Connotea and CiteULike, provide researchers with a means to organize personal collections of online references with keywords (tags) and to share these collections with others. One of the side-effects of the operation of these systems is the generation of large, publicly accessible metadata repositories describing the resources in the collections. In light of the well-known expansion of information in the life sciences and the need for metadata to enhance its value, these repositories present a potentially valuable new resource for application developers. Here we characterize the current contents of two scientifically relevant metadata repositories created through social tagging. This investigation helps to establish how such socially constructed metadata might be used as it stands currently and to suggest ways that new social tagging systems might be designed that would yield better aggregate products. We assessed the metadata that users of CiteULike and Connotea associated with citations in PubMed with the following metrics: coverage of the document space, density of metadata (tags) per document, rates of inter-annotator agreement, and rates of agreement with MeSH indexing. CiteULike and Connotea were very similar on all of the measurements. In comparison to PubMed, document coverage and per-document metadata density were much lower for the social tagging systems. Inter-annotator agreement within the social tagging systems and the agreement between the aggregated social tagging metadata and MeSH indexing was low though the latter could be increased through voting. The most promising uses of metadata from current academic social tagging repositories will be those that find ways to utilize the novel relationships between users, tags, and documents exposed through these systems. For more traditional kinds of indexing-based applications (such as keyword-based search) to benefit substantially from socially generated metadata in the life sciences, more documents need to be tagged and more tags are needed for each document. These issues may be addressed both by finding ways to attract more users to current systems and by creating new user interfaces that encourage more collectively useful individual tagging behaviour.

  8. Blockade of Tumor Cell TGF-Betas: A Strategy to Reverse Antiestrogen Resistance in Human Breast Cancer

    DTIC Science & Technology

    2002-01-01

    the TM- FKHRL1 construct exhibited exclusive nuclear localization Cell Cycle Analysis by Flow Cytometry of the HA-tagged mutant under any experimental...distribution as measured by flow cytometry (Figure 8A). ALS AND METHODS. Consistent with its antiapoptotic effect, these results, addi- tion of TGFI3... flow cytometry . Under these conditions more than 95% of selected cells expressed GFP at the time of experiments. Immunoblot Analysis. Cells were

  9. Design of a Humidity Sensor Tag for Passive Wireless Applications.

    PubMed

    Wu, Xiang; Deng, Fangming; Hao, Yong; Fu, Zhihui; Zhang, Lihua

    2015-10-07

    This paper presents a wireless humidity sensor tag for low-cost and low-power applications. The proposed humidity sensor tag, based on radio frequency identification (RFID) technology, was fabricated in a standard 0.18 μm complementary metal oxide semiconductor (CMOS) process. The top metal layer was deposited to form the interdigitated electrodes, which were then filled with polyimide as the humidity sensing layer. A two-stage rectifier adopts a dynamic bias-voltage generator to boost the effective gate-source voltage of the switches in differential-drive architecture, resulting in a flat power conversion efficiency curve. The capacitive sensor interface, based on phase-locked loop (PLL) theory, employs a simple architecture and can work with 0.5 V supply voltage. The measurement results show that humidity sensor tag achieves excellent linearity, hysteresis and stability performance. The total power-dissipation of the sensor tag is 2.5 μW, resulting in a maximum operating distance of 23 m under 4 W of radiation power of the RFID reader.

  10. Engineering Protein Hydrogels Using SpyCatcher-SpyTag Chemistry.

    PubMed

    Gao, Xiaoye; Fang, Jie; Xue, Bin; Fu, Linglan; Li, Hongbin

    2016-09-12

    Constructing hydrogels from engineered proteins has attracted significant attention within the material sciences, owing to their myriad potential applications in biomedical engineering. Developing efficient methods to cross-link tailored protein building blocks into hydrogels with desirable mechanical, physical, and functional properties is of paramount importance. By making use of the recently developed SpyCatcher-SpyTag chemistry, we successfully engineered protein hydrogels on the basis of engineered tandem modular elastomeric proteins. Our resultant protein hydrogels are soft but stable, and show excellent biocompatibility. As the first step, we tested the use of these hydrogels as a drug carrier, as well as in encapsulating human lung fibroblast cells. Our results demonstrate the robustness of the SpyCatcher-SpyTag chemistry, even when the SpyTag (or SpyCatcher) is flanked by folded globular domains. These results demonstrate that SpyCatcher-SpyTag chemistry can be used to engineer protein hydrogels from tandem modular elastomeric proteins that can find applications in tissue engineering, in fundamental mechano-biological studies, and as a controlled drug release vehicle.

  11. Design of a Humidity Sensor Tag for Passive Wireless Applications

    PubMed Central

    Wu, Xiang; Deng, Fangming; Hao, Yong; Fu, Zhihui; Zhang, Lihua

    2015-01-01

    This paper presents a wireless humidity sensor tag for low-cost and low-power applications. The proposed humidity sensor tag, based on radio frequency identification (RFID) technology, was fabricated in a standard 0.18 μm complementary metal oxide semiconductor (CMOS) process. The top metal layer was deposited to form the interdigitated electrodes, which were then filled with polyimide as the humidity sensing layer. A two-stage rectifier adopts a dynamic bias-voltage generator to boost the effective gate-source voltage of the switches in differential-drive architecture, resulting in a flat power conversion efficiency curve. The capacitive sensor interface, based on phase-locked loop (PLL) theory, employs a simple architecture and can work with 0.5 V supply voltage. The measurement results show that humidity sensor tag achieves excellent linearity, hysteresis and stability performance. The total power-dissipation of the sensor tag is 2.5 μW, resulting in a maximum operating distance of 23 m under 4 W of radiation power of the RFID reader. PMID:26457707

  12. Regulation of calcium-permeable TRPV2 channel by insulin in pancreatic beta-cells.

    PubMed

    Hisanaga, Etsuko; Nagasawa, Masahiro; Ueki, Kohjiro; Kulkarni, Rohit N; Mori, Masatomo; Kojima, Itaru

    2009-01-01

    Calcium-permeable cation channel TRPV2 is expressed in pancreatic beta-cells. We investigated regulation and function of TRPV2 in beta-cells. Translocation of TRPV2 was assessed in MIN6 cells and cultured mouse beta-cells by transfecting TRPV2 fused to green fluorescent protein or TRPV2 containing c-Myc tag in the extracellular domain. Calcium entry was assessed by monitoring fura-2 fluorescence. In MIN6 cells, TRPV2 was observed mainly in cytoplasm in an unstimulated condition. Addition of exogenous insulin induced translocation and insertion of TRPV2 to the plasma membrane. Consistent with these observations, insulin increased calcium entry, which was inhibited by tranilast, an inhibitor of TRPV2, or by knockdown of TRPV2 using shRNA. A high concentration of glucose also induced translocation of TRPV2, which was blocked by nefedipine, diazoxide, and somatostatin, agents blocking glucose-induced insulin secretion. Knockdown of the insulin receptor attenuated insulin-induced translocation of TRPV2. Similarly, the effect of insulin on TRPV2 translocation was not observed in a beta-cell line derived from islets obtained from a beta-cell-specific insulin receptor knockout mouse. Knockdown of TRPV2 or addition of tranilast significantly inhibited insulin secretion induced by a high concentration of glucose. Likewise, cell growth induced by serum and glucose was inhibited by tranilast or by knockdown of TRPV2. Finally, insulin-induced translocation of TRPV2 was observed in cultured mouse beta-cells, and knockdown of TRPV2 reduced insulin secretion induced by glucose. TRPV2 is regulated by insulin and is involved in the autocrine action of this hormone on beta-cells.

  13. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dowla, F

    Long-range, ultra-wideband (UWB), passive radio frequency (RF) tags are key components in Radio Frequency IDentification (RFID) system that will revolutionize inventory control and tracking applications. Unlike conventional, battery-operated (active) RFID tags, LLNL's small UWB tags, called 'UTag', operate at long range (up to 20 meters) in harsh, cluttered environments. Because they are battery-less (that is, passive), they have practically infinite lifetimes without human intervention, and they are lower in cost to manufacture and maintain than active RFID tags. These robust, energy-efficient passive tags are remotely powered by UWB radio signals, which are much more difficult to detect, intercept, and jammore » than conventional narrowband frequencies. The features of long range, battery-less, and low cost give UTag significant advantage over other existing RFID tags.« less

  14. Visualizing Sweetness: Increasingly Diverse Applications for Fluorescent-Tagged Glucose Bioprobes and Their Recent Structural Modifications

    PubMed Central

    Kim, Woong Hee; Lee, Jinho; Jung, Da-Woon; Williams, Darren R.

    2012-01-01

    Glucose homeostasis is a fundamental aspect of life and its dysregulation is associated with important diseases, such as cancer and diabetes. Traditionally, glucose radioisotopes have been used to monitor glucose utilization in biological systems. Fluorescent-tagged glucose analogues were initially developed in the 1980s, but it is only in the past decade that their use as a glucose sensor has increased significantly. These analogues were developed for monitoring glucose uptake in blood cells, but their recent applications include tracking glucose uptake by tumor cells and imaging brain cell metabolism. This review outlines the development of fluorescent-tagged glucose analogues, describes their recent structural modifications and discusses their increasingly diverse biological applications. PMID:22666073

  15. A method for the analysis of sugars in biological systems using reductive amination in combination with hydrophilic interaction chromatography and high resolution mass spectrometry.

    PubMed

    Bawazeer, Sami; Muhsen Ali, Ali; Alhawiti, Aliyah; Khalaf, Abedawn; Gibson, Colin; Tusiimire, Jonans; Watson, David G

    2017-05-01

    Separation of sugar isomers extracted from biological samples is challenging because of their natural occurrence as alpha and beta anomers and, in the case of hexoses, in their pyranose and furanose forms. A reductive amination method was developed for the tagging of sugars with the aim of it becoming part of a metabolomics work flow. The best separation of the common hexoses (glucose, fructose, mannose and galactose) was achieved when 2 H 5 -aniline was used as the tagging reagent in combination with separation on a ZICHILIC column. The method was used to tag a range of sugars including pentoses and uronic acids. The method was simple to perform and was able to improve both the separation of sugars and their response to electrospray ionisation. The method was applied to the profiling of sugars in urine where a number of hexose and pentose isomers could be observed. It was also applied to the quantification of sugars in post-mortem brain samples from three control samples and three samples from individuals who had suffered from bipolar disorder. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Direct and Indirect Visualization of Bacterial Effector Delivery into Diverse Plant Cell Types during Infection[OPEN

    PubMed Central

    Henry, Elizabeth; Jauneau, Alain; Deslandes, Laurent

    2017-01-01

    To cause disease, diverse pathogens deliver effector proteins into host cells. Pathogen effectors can inhibit defense responses, alter host physiology, and represent important cellular probes to investigate plant biology. However, effector function and localization have primarily been investigated after overexpression in planta. Visualizing effector delivery during infection is challenging due to the plant cell wall, autofluorescence, and low effector abundance. Here, we used a GFP strand system to directly visualize bacterial effectors delivered into plant cells through the type III secretion system. GFP is a beta barrel that can be divided into 11 strands. We generated transgenic Arabidopsis thaliana plants expressing GFP1-10 (strands 1 to 10). Multiple bacterial effectors tagged with the complementary strand 11 epitope retained their biological function in Arabidopsis and tomato (Solanum lycopersicum). Infection of plants expressing GFP1-10 with bacteria delivering GFP11-tagged effectors enabled direct effector detection in planta. We investigated the temporal and spatial delivery of GFP11-tagged effectors during infection with the foliar pathogen Pseudomonas syringae and the vascular pathogen Ralstonia solanacearum. Thus, the GFP strand system can be broadly used to investigate effector biology in planta. PMID:28600390

  17. Achievable Performance and Effective Interrogator Design for SAW RFID Sensor Tags

    NASA Technical Reports Server (NTRS)

    Barton, Richard J.

    2011-01-01

    For many NASA missions, remote sensing is a critical application that supports activities such as environmental monitoring, planetary science, structural shape and health monitoring, non-destructive evaluation, etc. The utility of the remote sensing devices themselves is greatly increased if they are passive that is, they do not require any on-board power supply such as batteries and if they can be identified uniquely during the sensor interrogation process. Additional passive sensor characteristics that enable greater utilization in space applications are small size and weight, long read ranges with low interrogator power, ruggedness, and operability in extreme environments (vacuum, extreme high/low temperature, high radiation, etc.) In this paper, we consider one very promising passive sensor technology, called surface acoustic wave (SAW) radio-frequency identification (RFID), that satisfies all of these criteria. Although SAW RFID tags have great potential for use in numerous space-based remote sensing applications, the limited collision resolution capability of current generation tags limits the performance in a cluttered sensing environment. That is, as more SAW-based sensors are added to the environment, numerous tag responses are superimposed at the receiver and decoding all or even a subset of the telemetry becomes increasingly difficult. Background clutter generated by reflectors other than the sensors themselves is also a problem, as is multipath interference and signal distortion, but the limiting factor in many remote sensing applications can be expected to be tag mutual interference. This problem may be greatly mitigated by proper design of the SAW tag waveform, but that remains an open research problem, and in the meantime, several other related questions remain to be answered including: What are the fundamental relationships between tag parameters such as bit-rate, time-bandwidth-product, SNR, and achievable collision resolution? What are the differences in optimal or near-optimal interrogator designs between noise-limited environments and interference-limited environments? What are the performance characteristics of different interrogator designs in term of parameters such as transmitter power level, range, and number of interfering tags? In this paper, we present the results of a research effort aimed at providing at least partial answers to all of these questions.

  18. Design and Optimization of Passive UHF RFID Tag Antenna for Mounting on or inside Material Layers

    NASA Astrophysics Data System (ADS)

    Shao, Shuai

    There is great desire to employ passive UHF RFID tags for inventory tracking and sensing in a diversity of applications and environments. Owing to its battery-free operation, non-line-of sight detection, low cost, long read range and small form factor, each year billions of RFID tags are being deployed in retail, logistics, manufacturing, biomedical inventories, among many other applications. However, the performance of these RFID systems has not met expectations. This is because a tag's performance deteriorates significantly when mounted on or inside arbitrary materials. The tag antenna is optimized only for a given type of material at a certain location of placement, and detuning takes place when attached to or embedded in materials with dielectric properties outside the design range. Thereby, different customized tags may be needed for identifying objects even within the same class of products. This increases the overall cost of the system. Furthermore, conventional copper foil-based RFID tag antennas are prone to metal fatigue and wear, and cannot survive hostile environments where antennas could be deformed by external forces and failures occur. Therefore, it is essential to understand the interaction between the antenna and the material in the vicinity of the tag, and design general purpose RFID tag antennas possessing excellent electrical performance as well as robust mechanical structure. A particularly challenging application addressed here is designing passive RFID tag antennas for automotive tires. Tires are composed of multiple layers of rubber with different dielectric properties and thicknesses. Furthermore, metallic plies are embedded in the sidewalls and steel belts lie beneath the tread to enforce mechanical integrity. To complicate matters even more, a typical tire experiences a 10% stretching during the construction process. This dissertation focuses on intuitively understanding the interaction between the antenna and the material in the proximity and designing broad band and mechanically robust RFID tag antennas for elastic materials. As a first step, the effects of dielectric materials on an antenna's impedance match and radiation pattern are investigated. The detuning effect is quantified based on the theoretical frequency scaling and effective permittivity of a dielectric material of finite thickness. Using simple formulas, the operational range of a tag can be predicted without intensive full-wave simulations of different materials. Next, a spectral domain Green's function is applied to compute the antenna pattern when the tag is mounted on or inside a layered medium. The optimal placement of the tag is found based on the focusing effect that the material has on the gain pattern of the antenna. For tires, the steel ply in the sidewall of a tire looks like a periodic wire grating. The performance of an antenna placed close to a wire grating is predicted using Floquet theory. The results indicate that steel plies embedded in the tire can be utilized as a reflector to further focus the gain pattern and increase the read range of a tag. Using these design tools and theoretical analysis, several broadband RFID tag antennas are designed for multi-layered materials. A novel stretchable conductive textile (E-fiber) based tag antenna is also developed for placement in elastic materials. Prototype antennas are fabricated and embedded in a tire during the tire manufacturing process. Experimental results indicate that tags with the new antennas achieve significant improvement compared with commercially available tags.

  19. An Ultra-Low-Power RFID/NFC Frontend IC Using 0.18 μm CMOS Technology for Passive Tag Applications.

    PubMed

    Bhattacharyya, Mayukh; Gruenwald, Waldemar; Jansen, Dirk; Reindl, Leonhard; Aghassi-Hagmann, Jasmin

    2018-05-07

    Battery-less passive sensor tags based on RFID or NFC technology have achieved much popularity in recent times. Passive tags are widely used for various applications like inventory control or in biotelemetry. In this paper, we present a new RFID/NFC frontend IC (integrated circuit) for 13.56 MHz passive tag applications. The design of the frontend IC is compatible with the standard ISO 15693/NFC 5. The paper discusses the analog design part in details with a brief overview of the digital interface and some of the critical measured parameters. A novel approach is adopted for the demodulator design, to demodulate the 10% ASK (amplitude shift keying) signal. The demodulator circuit consists of a comparator designed with a preset offset voltage. The comparator circuit design is discussed in detail. The power consumption of the bandgap reference circuit is used as the load for the envelope detection of the ASK modulated signal. The sub-threshold operation and low-supply-voltage are used extensively in the analog design—to keep the power consumption low. The IC was fabricated using 0.18 μ m CMOS technology in a die area of 1.5 mm × 1.5 mm and an effective area of 0.7 m m 2 . The minimum supply voltage desired is 1.2 V, for which the total power consumption is 107 μ W. The analog part of the design consumes only 36 μ W, which is low in comparison to other contemporary passive tags ICs. Eventually, a passive tag is developed using the frontend IC, a microcontroller, a temperature and a pressure sensor. A smart NFC device is used to readout the sensor data from the tag employing an Android-based application software. The measurement results demonstrate the full passive operational capability. The IC is suitable for low-power and low-cost industrial or biomedical battery-less sensor applications. A figure-of-merit (FOM) is proposed in this paper which is taken as a reference for comparison with other related state-of-the-art researches.

  20. An Ultra-Low-Power RFID/NFC Frontend IC Using 0.18 μm CMOS Technology for Passive Tag Applications

    PubMed Central

    Gruenwald, Waldemar; Jansen, Dirk; Aghassi-Hagmann, Jasmin

    2018-01-01

    Battery-less passive sensor tags based on RFID or NFC technology have achieved much popularity in recent times. Passive tags are widely used for various applications like inventory control or in biotelemetry. In this paper, we present a new RFID/NFC frontend IC (integrated circuit) for 13.56 MHz passive tag applications. The design of the frontend IC is compatible with the standard ISO 15693/NFC 5. The paper discusses the analog design part in details with a brief overview of the digital interface and some of the critical measured parameters. A novel approach is adopted for the demodulator design, to demodulate the 10% ASK (amplitude shift keying) signal. The demodulator circuit consists of a comparator designed with a preset offset voltage. The comparator circuit design is discussed in detail. The power consumption of the bandgap reference circuit is used as the load for the envelope detection of the ASK modulated signal. The sub-threshold operation and low-supply-voltage are used extensively in the analog design—to keep the power consumption low. The IC was fabricated using 0.18 μm CMOS technology in a die area of 1.5 mm × 1.5 mm and an effective area of 0.7 mm2. The minimum supply voltage desired is 1.2 V, for which the total power consumption is 107 μW. The analog part of the design consumes only 36 μW, which is low in comparison to other contemporary passive tags ICs. Eventually, a passive tag is developed using the frontend IC, a microcontroller, a temperature and a pressure sensor. A smart NFC device is used to readout the sensor data from the tag employing an Android-based application software. The measurement results demonstrate the full passive operational capability. The IC is suitable for low-power and low-cost industrial or biomedical battery-less sensor applications. A figure-of-merit (FOM) is proposed in this paper which is taken as a reference for comparison with other related state-of-the-art researches. PMID:29735939

  1. Subcellular glucose exposure biases the spatial distribution of insulin granules in single pancreatic beta cells.

    PubMed

    Terao, Kyohei; Gel, Murat; Okonogi, Atsuhito; Fuke, Ariko; Okitsu, Teru; Tada, Takashi; Suzuki, Takaaki; Nagamatsu, Shinya; Washizu, Masao; Kotera, Hidetoshi

    2014-02-18

    In living tissues, a cell is exposed to chemical substances delivered partially to its surface. Such a heterogeneous chemical environment potentially induces cell polarity. To evaluate this effect, we developed a microfluidic device that realizes spatially confined delivery of chemical substances at subcellular resolution. Our microfluidic device allows simple setup and stable operation for over 4 h to deliver chemicals partially to a single cell. Using the device, we showed that subcellular glucose exposure triggers an intracellular [Ca(2+)] change in the β-cells. In addition, the imaging of a cell expressing GFP-tagged insulin showed that continuous subcellular exposure to glucose biased the spatial distribution of insulin granules toward the site where the glucose was delivered. Our approach illustrates an experimental technique that will be applicable to many biological experiments for imaging the response to subcellular chemical exposure and will also provide new insights about the development of polarity of β-cells.

  2. Subcellular glucose exposure biases the spatial distribution of insulin granules in single pancreatic beta cells

    PubMed Central

    Terao, Kyohei; Gel, Murat; Okonogi, Atsuhito; Fuke, Ariko; Okitsu, Teru; Tada, Takashi; Suzuki, Takaaki; Nagamatsu, Shinya; Washizu, Masao; Kotera, Hidetoshi

    2014-01-01

    In living tissues, a cell is exposed to chemical substances delivered partially to its surface. Such a heterogeneous chemical environment potentially induces cell polarity. To evaluate this effect, we developed a microfluidic device that realizes spatially confined delivery of chemical substances at subcellular resolution. Our microfluidic device allows simple setup and stable operation for over 4 h to deliver chemicals partially to a single cell. Using the device, we showed that subcellular glucose exposure triggers an intracellular [Ca2+] change in the β-cells. In addition, the imaging of a cell expressing GFP-tagged insulin showed that continuous subcellular exposure to glucose biased the spatial distribution of insulin granules toward the site where the glucose was delivered. Our approach illustrates an experimental technique that will be applicable to many biological experiments for imaging the response to subcellular chemical exposure and will also provide new insights about the development of polarity of β-cells. PMID:24535122

  3. IM-TORNADO: A Tool for Comparison of 16S Reads from Paired-End Libraries

    PubMed Central

    Jeraldo, Patricio; Kalari, Krishna; Chen, Xianfeng; Bhavsar, Jaysheel; Mangalam, Ashutosh; White, Bryan; Nelson, Heidi; Kocher, Jean-Pierre; Chia, Nicholas

    2014-01-01

    Motivation 16S rDNA hypervariable tag sequencing has become the de facto method for accessing microbial diversity. Illumina paired-end sequencing, which produces two separate reads for each DNA fragment, has become the platform of choice for this application. However, when the two reads do not overlap, existing computational pipelines analyze data from read separately and underutilize the information contained in the paired-end reads. Results We created a workflow known as Illinois Mayo Taxon Organization from RNA Dataset Operations (IM-TORNADO) for processing non-overlapping reads while retaining maximal information content. Using synthetic mock datasets, we show that the use of both reads produced answers with greater correlation to those from full length 16S rDNA when looking at taxonomy, phylogeny, and beta-diversity. Availability and Implementation IM-TORNADO is freely available at http://sourceforge.net/projects/imtornado and produces BIOM format output for cross compatibility with other pipelines such as QIIME, mothur, and phyloseq. PMID:25506826

  4. Store operation with conditional push of a tag value to a queue

    DOEpatents

    Heidelberger, Philip; Steinmacher-Burow, Burkhard

    2015-07-28

    According to one embodiment, a method for a store operation with a conditional push of a tag value to a queue is provided. The method includes configuring a queue that is accessible by an application, setting a value at an address in a memory device including a memory and a controller, receiving a request for an operation using the value at the address and performing the operation. The method also includes the controller writing a result of the operation to the address, thus changing the value at the address, the controller determining if the result of the operation meets a condition and the controller pushing a tag value to the queue based on the condition being met, where the tag value in the queue indicates to the application that the condition is met.

  5. Postprandial lipid responses to an alpha-linolenic acid-rich oil, olive oil and butter in women: A randomized crossover trial

    PubMed Central

    2011-01-01

    Background Postprandial lipaemia varies with gender and the composition of dietary fat due to the partitioning of fatty acids between beta-oxidation and incorporation into triacylglycerols (TAGs). Increasing evidence highlights the importance of postprandial measurements to evaluate atherogenic risk. Postprandial effects of alpha-linolenic acid (ALA) in women are poorly characterized. We therefore studied the postprandial lipid response of women to an ALA-rich oil in comparison with olive oil and butter, and characterized the fatty acid composition of total lipids, TAGs, and non-esterified fatty acids (NEFAs) in plasma. Methods A randomized crossover design (n = 19) was used to compare the postprandial effects of 3 meals containing 35 g fat. Blood samples were collected at regular intervals for 7 h. Statistical analysis was carried out with ANOVA (significant difference = P < 0.05). Results No significant difference was seen in incremental area under the curve (iAUC) plasma-TAG between the meals. ALA and oleic acid levels were significantly increased in plasma after ALA-rich oil and olive oil meals, respectively. Palmitic acid was significantly increased in plasma-TAG after the butter meal. The ratios of 18:2 n-6 to18:3 n-3 in plasma-TAGs, three and seven hours after the ALA-rich oil meal, were 1.5 and 2.4, respectively. The corresponding values after the olive oil meal were: 13.8 and 16.9; and after the butter meal: 9.0 and 11.6. Conclusions The postprandial p-TAG and NEFA response in healthy pre-menopausal women was not significantly different after the intake of an ALA-rich oil, olive oil and butter. The ALA-rich oil significantly affected different plasma lipid fractions and improved the ratio of n-6 to n-3 fatty acids several hours postprandially. PMID:21711508

  6. Purification of FLAG-tagged Secreted Proteins from Mammalian Cells

    PubMed Central

    Itakura, Eisuke; Chen, Changchun; de Bono, Mario

    2017-01-01

    This protocol describes a method for purifying glycosylated FLAG-tagged secreted proteins with disulfide bonds from mammalian cells. The purified products can be used for various applications, such as ligand binding assays. PMID:29075655

  7. A scale space based algorithm for automated segmentation of single shot tagged MRI of shearing deformation.

    PubMed

    Sprengers, Andre M J; Caan, Matthan W A; Moerman, Kevin M; Nederveen, Aart J; Lamerichs, Rolf M; Stoker, Jaap

    2013-04-01

    This study proposes a scale space based algorithm for automated segmentation of single-shot tagged images of modest SNR. Furthermore the algorithm was designed for analysis of discontinuous or shearing types of motion, i.e. segmentation of broken tag patterns. The proposed algorithm utilises non-linear scale space for automatic segmentation of single-shot tagged images. The algorithm's ability to automatically segment tagged shearing motion was evaluated in a numerical simulation and in vivo. A typical shearing deformation was simulated in a Shepp-Logan phantom allowing for quantitative evaluation of the algorithm's success rate as a function of both SNR and the amount of deformation. For a qualitative in vivo evaluation tagged images showing deformations in the calf muscles and eye movement in a healthy volunteer were acquired. Both the numerical simulation and the in vivo tagged data demonstrated the algorithm's ability for automated segmentation of single-shot tagged MR provided that SNR of the images is above 10 and the amount of deformation does not exceed the tag spacing. The latter constraint can be met by adjusting the tag delay or the tag spacing. The scale space based algorithm for automatic segmentation of single-shot tagged MR enables the application of tagged MR to complex (shearing) deformation and the processing of datasets with relatively low SNR.

  8. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rowe, Nathan C; Younkin, James R; Pickett, Chris A

    Radio-Frequency (RF) devices have revolutionized many aspects of modern industrial processes. RF technology can enable wireless communication for tag identification, sensor communication, and asset tracking. Radio-frequency identification (RFID) is a technology that utilizes wireless communication to interrogate and identify an electronic tag attached to an item in order to identify the item. The technology can come in many forms: passive or active tags, low to ultra-wideband frequencies, small paper-thin tags to brick-sized units, and simple tags or highly integrated sensor packages. RF technology, and specifically RFID, has been applied widely in commercial markets for inventory, supply chain management, and assetmore » tracking. Several recent studies have demonstrated the safeguards benefits of utilizing RFID versus conventional inventory tagging methods for tracking nuclear material. These studies have indicated that the RF requirements for safeguards functions are more stringent than the RF requirements for other inventory tracking and accounting applications. Additionally, other requirements must be addressed, including environmental and operating conditions, authentication, and tag location and attachment. Facility restrictions on radio spectrum, method of tag attachment, and sensitivity of the data collected impact the tag selection and system design. More important, the intended use of the system must be considered. The requirements for using RF to simply replace or supplement container identifiers such as bar codes that facilitate the inventory function will differ greatly from the requirements for deploying RF for unattended monitoring applications. Several studies have investigated these considerations to advance commercial RF devices for safeguards use, and a number of system concepts have been developed. This paper will provide an overview of past studies and current technologies, and will investigate the requirements, existing gaps, and several potential next steps for advancing RF techniques for safeguards use.« less

  9. Biotin-tagged proteins: Reagents for efficient ELISA-based serodiagnosis and phage display-based affinity selection

    PubMed Central

    Verma, Vaishali; Kaur, Charanpreet; Grover, Payal; Gupta, Amita

    2018-01-01

    The high-affinity interaction between biotin and streptavidin has opened avenues for using recombinant proteins with site-specific biotinylation to achieve efficient and directional immobilization. The site-specific biotinylation of proteins carrying a 15 amino acid long Biotin Acceptor Peptide tag (BAP; also known as AviTag) is effected on a specific lysine either by co-expressing the E. coli BirA enzyme in vivo or by using purified recombinant E. coli BirA enzyme in the presence of ATP and biotin in vitro. In this paper, we have designed a T7 promoter-lac operator-based expression vector for rapid and efficient cloning, and high-level cytosolic expression of proteins carrying a C-terminal BAP tag in E. coli with TEV protease cleavable N-terminal deca-histidine tag, useful for initial purification. Furthermore, a robust three-step purification pipeline integrated with well-optimized protocols for TEV protease-based H10 tag removal, and recombinant BirA enzyme-based site-specific in vitro biotinylation is described to obtain highly pure biotinylated proteins. Most importantly, the paper demonstrates superior sensitivities in indirect ELISA with directional and efficient immobilization of biotin-tagged proteins on streptavidin-coated surfaces in comparison to passive immobilization. The use of biotin-tagged proteins through specific immobilization also allows more efficient selection of binders from a phage-displayed naïve antibody library. In addition, for both these applications, specific immobilization requires much less amount of protein as compared to passive immobilization and can be easily multiplexed. The simplified strategy described here for the production of highly pure biotin-tagged proteins will find use in numerous applications, including those, which may require immobilization of multiple proteins simultaneously on a solid surface. PMID:29360877

  10. Biotin-tagged proteins: Reagents for efficient ELISA-based serodiagnosis and phage display-based affinity selection.

    PubMed

    Verma, Vaishali; Kaur, Charanpreet; Grover, Payal; Gupta, Amita; Chaudhary, Vijay K

    2018-01-01

    The high-affinity interaction between biotin and streptavidin has opened avenues for using recombinant proteins with site-specific biotinylation to achieve efficient and directional immobilization. The site-specific biotinylation of proteins carrying a 15 amino acid long Biotin Acceptor Peptide tag (BAP; also known as AviTag) is effected on a specific lysine either by co-expressing the E. coli BirA enzyme in vivo or by using purified recombinant E. coli BirA enzyme in the presence of ATP and biotin in vitro. In this paper, we have designed a T7 promoter-lac operator-based expression vector for rapid and efficient cloning, and high-level cytosolic expression of proteins carrying a C-terminal BAP tag in E. coli with TEV protease cleavable N-terminal deca-histidine tag, useful for initial purification. Furthermore, a robust three-step purification pipeline integrated with well-optimized protocols for TEV protease-based H10 tag removal, and recombinant BirA enzyme-based site-specific in vitro biotinylation is described to obtain highly pure biotinylated proteins. Most importantly, the paper demonstrates superior sensitivities in indirect ELISA with directional and efficient immobilization of biotin-tagged proteins on streptavidin-coated surfaces in comparison to passive immobilization. The use of biotin-tagged proteins through specific immobilization also allows more efficient selection of binders from a phage-displayed naïve antibody library. In addition, for both these applications, specific immobilization requires much less amount of protein as compared to passive immobilization and can be easily multiplexed. The simplified strategy described here for the production of highly pure biotin-tagged proteins will find use in numerous applications, including those, which may require immobilization of multiple proteins simultaneously on a solid surface.

  11. Mobilization of lipid reserves during germination of oat (Avena sativa L.), a cereal rich in endosperm oil.

    PubMed

    Leonova, Svetlana; Grimberg, Asa; Marttila, Salla; Stymne, Sten; Carlsson, Anders S

    2010-06-01

    Since the cereal endosperm is a dead tissue in the mature grain, beta-oxidation is not possible there. This raises the question about the use of the endosperm oil in cereal grains during germination. In this study, mobilization of lipids in different tissues of germinating oat grains was analysed using thin-layer and gas chromatography. The data imply that the oat endosperm oil [triacylglycerol (TAG)] is not a dead-end product as it was absorbed by the scutellum, either as free fatty acids (FFAs) released from TAG or as intact TAG immediately degraded to FFAs. These data were supported by light and transmission electron microscopy (LM and TEM) studies where close contact between endosperm lipid droplets and the scutellum was observed. The appearance of the fused oil in the oat endosperm changed into oil droplets during germination in areas close to the aleurone and the scutellar epithelium. However, according to the data obtained by TEM these oil droplets are unlikely to be oil bodies surrounded by oleosins. Accumulation of FFA pools in the embryo suggested further transport of FFAs from the scutellum. Noticeably high levels of TAG were also accumulated in the embryo but were not synthesized by re-esterification from imported FFAs. Comparison between two oat cultivars with different amounts of oil and starch in the endosperm suggests that an increased oil to starch ratio in oat grains does not significantly impact the germination process.

  12. Tag-to-Tag Interference Suppression Technique Based on Time Division for RFID.

    PubMed

    Khadka, Grishma; Hwang, Suk-Seung

    2017-01-01

    Radio-frequency identification (RFID) is a tracking technology that enables immediate automatic object identification and rapid data sharing for a wide variety of modern applications using radio waves for data transmission from a tag to a reader. RFID is already well established in technical areas, and many companies have developed corresponding standards and measurement techniques. In the construction industry, effective monitoring of materials and equipment is an important task, and RFID helps to improve monitoring and controlling capabilities, in addition to enabling automation for construction projects. However, on construction sites, there are many tagged objects and multiple RFID tags that may interfere with each other's communications. This reduces the reliability and efficiency of the RFID system. In this paper, we propose an anti-collision algorithm for communication between multiple tags and a reader. In order to suppress interference signals from multiple neighboring tags, the proposed algorithm employs the time-division (TD) technique, where tags in the interrogation zone are assigned a specific time slot so that at every instance in time, a reader communicates with tags using the specific time slot. We present representative computer simulation examples to illustrate the performance of the proposed anti-collision technique for multiple RFID tags.

  13. New Technique for Barium Daughter Ion Identification in a Liquid Xe-136 Double Beta Decay Experiment

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fairbank, William

    2016-06-08

    This work addresses long-standing issues of fundamental interest in elementary particle physics. The most important outcome of this work is a new limit on neutrinoless double beta decay. This is an extremely rare and long-sought-after type of radioactive decay. If discovered, it would require changes in the standard model of the elementary constituents of matter, and would prove that neutrinos and antineutrinos are the same, a revolutionary concept in particle physics. Neutrinos are major components of the matter in the universe that are so small and so weakly interacting with other matter that their masses have not yet been discovered.more » A discovery of neutrinoless double beta decay could help determine the neutrino masses. An important outcome of the work on this project was the Colorado State University role in operating the EXO-200 neutrinoless double beta decay experiment and in analysis of the data from this experiment. One type of double beta decay of the isotope 136Xe, the two-neutrino variety, was discovered in this work. Although the other type of double beta decay, the neutrinoless variety, was not yet discovered in this work, a world’s best sensitivity of 1.9x10 25 year half-life was obtained. This result rules out a previous claim of a positive result in a different isotope. This work also establishes that the masses of the neutrinos are less than one millionth of that of electrons. A unique EXO-200 analysis, in which the CSU group had a leading role, has established for the first time ever in a liquid noble gas the fraction of daughter atoms from alpha and beta decay that are ionized. This result has important impact on other pending studies, including nucleon decay and barium tagging. Novel additional discoveries include multiphoton ionization of liquid xenon with UV pulsed lasers, which may find application in calibration of future noble liquid detectors, and studies of association and dissociation reactions of Ba + ions in gaseous xenon. Through this project, we are substantially closer to demonstrating “barium tagging”, i.e., detection of single daughter 136Ba atoms from 136Xe double beta decay. Milestones achieved include obtaining spectra of small numbers of Ba atoms and cryoprobe advances toward trapping single 136Ba atoms in solid xenon and probe extraction for detection. One of the other benefits to society is the training of six Ph.D. students in a variety of state-of-the-art technologies, half under primary support of this grant and half with partial support, with four finishing their Ph.D. degrees and two well on their way.« less

  14. Search for new physics in a precise 20F beta spectrum shape measurement

    NASA Astrophysics Data System (ADS)

    George, Elizabeth; Voytas, Paul; Chuna, Thomas; Naviliat-Cuncic, Oscar; Gade, Alexandra; Hughes, Max; Huyan, Xueying; Liddick, Sean; Minamisono, Kei; Paulauskas, Stanley; Weisshaar, Dirk; Ban, Gilles; Flechard, Xavier; Lienard, Etienne

    2015-10-01

    We are carrying out a measurement of the shape of the energy spectrum of β particles from 20F decay. We aim to achieve a relative precision below 3%, representing an order of magnitude improvement compared to previous experiments. This level of precision will enable a test of the so-called strong form of the conserved vector current (CVC) hypothesis, and should also enable us to place competitive limits on the contributions of exotic tensor couplings in beta decay. In order to control systematic effects, we are using a technique that takes advantage of high energy radioactive beams at the NSCL to implant the decaying nuclei in a scintillation detector deep enough that the emitted beta particles cannot escape. The β-particle energy is measured with the implantation detector after switching off the beam implantation. Ancillary detectors are used to tag the 1.633-MeV γ-rays following the β decay for coincidence measurements in order to reduce backgrounds. We will give an overview and report on the status of the experiment.

  15. A review of tags anti-collision and localization protocols in RFID networks.

    PubMed

    Ullah, S; Alsalih, W; Alsehaim, A; Alsadhan, N

    2012-12-01

    Radio Frequency IDentification (RFID) has allowed the realization of ubiquitous tracking and monitoring of physical objects wirelessly with minimum human interactions. It plays a key role in a wide range of applications including asset tracking, contactless payment, access control, transportation and logistics, and other industrial applications. On the other side, RFID systems face several technical challenges that need to be overcome in order to achieve their potential benefits; tags collisions and localization of tagged objects are two important challenges. Numerous anti-collision and localization protocols have been proposed to address these challenges. This paper reviews the state-of-art tags' anti-collision and localization protocols, and provides a deep insight into technical issues of these protocols. The probabilistic and deterministic anti-collision protocols are critically studied and compared in terms of different parameters. We further review distance estimation, scene analysis, and proximity localization schemes and provide useful suggestions. We also introduce a new hybrid direction that utilizes power control to spatially partition the interrogation range of a reader for more efficient anti-collision and localization. Finally, we present the applications of RFID systems in healthcare sectors.

  16. Antibiotic use during the intracoelomic implantation of electronic tags into fish

    USGS Publications Warehouse

    Mulcahy, D.M.

    2011-01-01

    The use of antibiotics, in particular, the use of a single dose of antibiotics during electronic tag implantation is of unproven value, and carries with it the potential for the development of antibiotic resistance in bacteria and the alteration of the immune response of the fish. Antibiotic use during electronic tag implantation must conform to relevant drug laws and regulations in the country where work is being done, including the requirements for withdrawal times before human consumption is a possibility. Currently, the choice of antibiotics (most often tetracycline or oxytetracycline) and the use of a single dose of the drug are decisions made without knowledge of the basic need for antibiotic usage and of the bacteria involved in infections that occur following electronic tag implantation. Correct perioperative use of an antibiotic is to apply the drug to the animal before surgery begins, to assure serum and tissue levels of the drug are adequate before the incision is made. However, the most common perioperative application of antibiotics during implantation of an electronic tag is to delay the administration of the drug, injecting it into the coelom after the electronic tag is inserted, just prior to closure of the incision. There is little empirical evidence that the present application of antibiotics in fish being implanted with electronic tags is of value. Improvements should first be made to surgical techniques, especially the use of aseptic techniques and sterilized instruments and electronic tags, before resorting to antibiotics. ?? 2010 Springer Science+Business Media B.V.(outside the USA).

  17. Learning to rank image tags with limited training examples.

    PubMed

    Songhe Feng; Zheyun Feng; Rong Jin

    2015-04-01

    With an increasing number of images that are available in social media, image annotation has emerged as an important research topic due to its application in image matching and retrieval. Most studies cast image annotation into a multilabel classification problem. The main shortcoming of this approach is that it requires a large number of training images with clean and complete annotations in order to learn a reliable model for tag prediction. We address this limitation by developing a novel approach that combines the strength of tag ranking with the power of matrix recovery. Instead of having to make a binary decision for each tag, our approach ranks tags in the descending order of their relevance to the given image, significantly simplifying the problem. In addition, the proposed method aggregates the prediction models for different tags into a matrix, and casts tag ranking into a matrix recovery problem. It introduces the matrix trace norm to explicitly control the model complexity, so that a reliable prediction model can be learned for tag ranking even when the tag space is large and the number of training images is limited. Experiments on multiple well-known image data sets demonstrate the effectiveness of the proposed framework for tag ranking compared with the state-of-the-art approaches for image annotation and tag ranking.

  18. The ATLAS TAGS database distribution and management - Operational challenges of a multi-terabyte distributed database

    NASA Astrophysics Data System (ADS)

    Viegas, F.; Malon, D.; Cranshaw, J.; Dimitrov, G.; Nowak, M.; Nairz, A.; Goossens, L.; Gallas, E.; Gamboa, C.; Wong, A.; Vinek, E.

    2010-04-01

    The TAG files store summary event quantities that allow a quick selection of interesting events. This data will be produced at a nominal rate of 200 Hz, and is uploaded into a relational database for access from websites and other tools. The estimated database volume is 6TB per year, making it the largest application running on the ATLAS relational databases, at CERN and at other voluntary sites. The sheer volume and high rate of production makes this application a challenge to data and resource management, in many aspects. This paper will focus on the operational challenges of this system. These include: uploading the data from files to the CERN's and remote sites' databases; distributing the TAG metadata that is essential to guide the user through event selection; controlling resource usage of the database, from the user query load to the strategy of cleaning and archiving of old TAG data.

  19. Diazirines as Potential Molecular Imaging Tags: Probing the Requirements for Efficient and Long-Lived SABRE-Induced Hyperpolarization.

    PubMed

    Shen, Kun; Logan, Angus W J; Colell, Johannes F P; Bae, Junu; Ortiz, Gerardo X; Theis, Thomas; Warren, Warren S; Malcolmson, Steven J; Wang, Qiu

    2017-09-25

    Diazirines are an attractive class of potential molecular tags for magnetic resonance imaging owing to their biocompatibility and ease of incorporation into a large variety of molecules. As recently reported, 15 N 2 -diazirine can be hyperpolarized by the SABRE-SHEATH method, sustaining both singlet and magnetization states, thus offering a path to long-lived polarization storage. Herein, we show the generality of this approach by illustrating that the diazirine tag alone is sufficient for achieving excellent signal enhancements with long-lasting polarization. Our investigations reveal the critical role of Lewis basic additives, including water, on achieving SABRE-promoted hyperpolarization. The application of this strategy to a 15 N 2 -diazirine-containing choline derivative demonstrates the potential of 15 N 2 -diazirines as molecular imaging tags for biomedical applications. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Production of recombinant human beta2-microglobulin for scintigraphic diagnosis of amyloidosis in uremia and hemodialysis.

    PubMed

    Linke, R P; Schäeffer, J; Gielow, P; Lindner, P; Lottspeich, F; Plückthun, A; Weiss, E H

    2000-02-01

    Amyloid of beta2-microglobulin (beta2m) origin can be diagnosed using 131I-radiolabelled-beta2m scintigraphy in patients with uremia and hemodialysis treatment. As the tracer beta2m is isolated from another patient, it carries the common risks, including viral infections such as Hepatitis B, C and HIV, which are associated with human plasma products. In order to exclude these risks we have produced recombinant human beta2m (rhbeta2m) in Escherichia coli. The expression vector pASK40DeltaLbeta2m(His)5 contains a C-terminal (His)5-tag for purification via immobilized metal ion affinity chromatography (IMAC). Size exclusion chromatography on a Superose 12 column represents the second step of purification. The isolated rhbeta2mH5 reacted in an immunochemically identical manner to native human beta2m, and showed a single band of approximately 11.8 kDa in Western blot analysis and revealed a single spot in two-dimensional gel electrophoresis. Mass spectrometry analysis revealed a single peak at the expected molecular mass of 12 415.8 Da. Uniformity was further proven by crystallization and N-terminal amino-acid sequence analysis. The rhbeta2mH5 protein was then produced under conditions that allow the intravenous use in humans. Intraveneously applied indium-111-labelled rhbeta2mH5 was monitored in hemodialysed patients with and without known beta2m-amyloidosis. The tracer was localized specifically to particular areas known to contain amyloid. Thus, this rhbeta2mH5 preparation is suitable for detecting amyloid-containing organs of the beta2m-class in vivo and fulfils the requirements of a tracer for common use. Finally, the use of indium-111 instead of iodine-131 has reduced the radioactive load and resulted in higher resolution.

  1. 20F beta spectrum shape and weak interaction tests

    NASA Astrophysics Data System (ADS)

    Voytas, Paul; George, Elizabeth; Chuna, Thomas; Naviliat-Cuncic, Oscar; Hughes, Max; Huyan, Xueying; Minamisono, Kei; Paulauskas, Stanley

    2016-09-01

    Precision measurements of the shape of beta spectra can test our understanding of the weak interaction. We are carrying out a measurement of the shape of the energy spectrum of β particles from 20F decay. The primary motivation is to test the so-called strong form of the conserved vector current (CVC) hypothesis. The measurement should also enable us to place competitive limits on the contributions of exotic tensor couplings in beta decay. We aim to achieve a relative precision better than 3% on the linear contribution to the shape. This represents an order of magnitude improvement compared to previous experiments in 20F. In order to control systematic effects, we are using a technique that takes advantage of high energy radioactive beams at the NSCL to implant the decaying nuclei in scintillation detectors deeply enough that the emitted beta particles cannot escape. The β-particle energy is measured with the implantation detector after switching off the implantation beam. Ancillary detectors are used to identify the 1.633-MeV γ-rays following the 20F β decay for coincidence measurements in order to tag the transition of interest and to reduce backgrounds. We report on the status of the analysis. Supported in part with Awards from the NSCL PAC and the National Science Foundation under Grant No. PHY-1506084.

  2. Measurement of the scintillation time spectra and pulse-shape discrimination of low-energy β and nuclear recoils in liquid argon with DEAP-1

    DOE PAGES

    Amaudruz, P. -A.; Batygov, M.; Beltran, B.; ...

    2016-09-17

    The DEAP-1 low-background liquid argon detector has been used to measure scintillation pulse shapes of beta decays and nuclear recoil events and to demonstrate the feasibility of pulse-shape discrimination down to an electron-equivalent energy of 20 keV ee. The relative intensities of singlet/triplet states in liquid argon have been measured as a function of energy between 15 and 500 keVee for both beta and nuclear recoils. Using a triple-coincidence tag we find the fraction of beta events that are misidentified as nuclear recoils to be less than 6 x 10 -8 between 43-86 keV ee and that the discrimination parametermore » agrees with a simple analytic model. The discrimination measurement is currently limited by nuclear recoils induced by cosmic-ray generated neutrons, and is expected to improve by operating the detector underground at SNOLAB. The analytic model predicts a beta misidentification fraction of 10 -10 for an electron-equivalent energy threshold of 20 keV ee. This reduction allows for a sensitive search for spin-independent scattering of WIMPs from 1000 kg of liquid argon with a WIMP-nucleon cross-section sensitivity of 10 -46 cm 2.« less

  3. Measurement of the scintillation time spectra and pulse-shape discrimination of low-energy β and nuclear recoils in liquid argon with DEAP-1

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Amaudruz, P. -A.; Batygov, M.; Beltran, B.

    The DEAP-1 low-background liquid argon detector has been used to measure scintillation pulse shapes of beta decays and nuclear recoil events and to demonstrate the feasibility of pulse-shape discrimination down to an electron-equivalent energy of 20 keV ee. The relative intensities of singlet/triplet states in liquid argon have been measured as a function of energy between 15 and 500 keVee for both beta and nuclear recoils. Using a triple-coincidence tag we find the fraction of beta events that are misidentified as nuclear recoils to be less than 6 x 10 -8 between 43-86 keV ee and that the discrimination parametermore » agrees with a simple analytic model. The discrimination measurement is currently limited by nuclear recoils induced by cosmic-ray generated neutrons, and is expected to improve by operating the detector underground at SNOLAB. The analytic model predicts a beta misidentification fraction of 10 -10 for an electron-equivalent energy threshold of 20 keV ee. This reduction allows for a sensitive search for spin-independent scattering of WIMPs from 1000 kg of liquid argon with a WIMP-nucleon cross-section sensitivity of 10 -46 cm 2.« less

  4. Metabolic engineering of biomass for high energy density: oilseed-like triacylglycerol yields from plant leaves

    PubMed Central

    Vanhercke, Thomas; El Tahchy, Anna; Liu, Qing; Zhou, Xue-Rong; Shrestha, Pushkar; Divi, Uday K; Ral, Jean-Philippe; Mansour, Maged P; Nichols, Peter D; James, Christopher N; Horn, Patrick J; Chapman, Kent D; Beaudoin, Frederic; Ruiz-López, Noemi; Larkin, Philip J; de Feyter, Robert C; Singh, Surinder P; Petrie, James R

    2014-01-01

    High biomass crops have recently attracted significant attention as an alternative platform for the renewable production of high energy storage lipids such as triacylglycerol (TAG). While TAG typically accumulates in seeds as storage compounds fuelling subsequent germination, levels in vegetative tissues are generally low. Here, we report the accumulation of more than 15% TAG (17.7% total lipids) by dry weight in Nicotiana tabacum (tobacco) leaves by the co-expression of three genes involved in different aspects of TAG production without severely impacting plant development. These yields far exceed the levels found in wild-type leaf tissue as well as previously reported engineered TAG yields in vegetative tissues of Arabidopsis thaliana and N. tabacum. When translated to a high biomass crop, the current levels would translate to an oil yield per hectare that exceeds those of most cultivated oilseed crops. Confocal fluorescence microscopy and mass spectrometry imaging confirmed the accumulation of TAG within leaf mesophyll cells. In addition, we explored the applicability of several existing oil-processing methods using fresh leaf tissue. Our results demonstrate the technical feasibility of a vegetative plant oil production platform and provide for a step change in the bioenergy landscape, opening new prospects for sustainable food, high energy forage, biofuel and biomaterial applications. PMID:24151938

  5. Pseudo-orthogonal frequency coded wireless SAW RFID temperature sensor tags.

    PubMed

    Saldanha, Nancy; Malocha, Donald C

    2012-08-01

    SAW sensors are ideal for various wireless, passive multi-sensor applications because they are small, rugged, radiation hard, and offer a wide range of material choices for operation over broad temperature ranges. The readable distance of a tag in a multi-sensor environment is dependent on the insertion loss of the device and the processing gain of the system. Single-frequency code division multiple access (CDMA) tags that are used in high-volume commercial applications must have universal coding schemes and large numbers of codes. The use of a large number of bits at the common center frequency to achieve sufficient code diversity in CDMA tags necessitates reflector banks with >30 dB loss. Orthogonal frequency coding is a spread-spectrum approach that employs frequency and time diversity to achieve enhanced tag properties. The use of orthogonal frequency coded (OFC) SAW tags reduces adjacent reflector interactions for low insertion loss, increased range, complex coding, and system processing gain. This work describes a SAW tag-sensor platform that reduces device loss by implementing long reflector banks with optimized spectral coding. This new pseudo-OFC (POFC) coding is defined and contrasted with the previously defined OFC coding scheme. Auto- and cross-correlation properties of the chips and their relation to reflectivity per strip and reflector length are discussed. Results at 250 MHz of 8-chip OFC and POFC SAW tags will be compared. The key parameters of insertion loss, cross-correlation, and autocorrelation of the two types of frequency-coded tags will be analyzed, contrasted, and discussed. It is shown that coded reflector banks can be achieved with near-zero loss and still maintain good coding properties. Experimental results and results predicted by the coupling of modes model are presented for varying reflector designs and codes. A prototype 915-MHz POFC sensor tag is used as a wireless temperature sensor and the results are shown.

  6. Implementing traceability using particle randomness-based textile printed tags

    NASA Astrophysics Data System (ADS)

    Agrawal, T. K.; Koehl, L.; Campagne, C.

    2017-10-01

    This article introduces a random particle-based traceability tag for textiles. The proposed tag not only act as a unique signature for the corresponding textile product but also possess the features such as easy to manufacture and hard to copy. It seeks applications in brand authentication and traceability in textile and clothing (T&C) supply chain. A prototype has been developed by screen printing process, in which micron-scale particles were mixed with the printing paste and printed on cotton fabrics to attain required randomness. To encode the randomness, the image of the developed tag was taken and analyzed using image processing. The randomness of the particles acts as a product key or unique signature which is required to decode the tag. Finally, washing and abrasion resistance tests were conducted to check the durability of the printed tag.

  7. Application of XML in DICOM

    NASA Astrophysics Data System (ADS)

    You, Xiaozhen; Yao, Zhihong

    2005-04-01

    As a standard of communication and storage for medical digital images, DICOM has been playing a very important role in integration of hospital information. In DICOM, tags are expressed by numbers, and only standard data elements can be shared by looking up Data Dictionary while private tags can not. As such, a DICOM file's readability and extensibility is limited. In addition, reading DICOM files needs special software. In our research, we introduced XML into DICOM, defining an XML-based DICOM special transfer format, XML-DCM, a DICOM storage format, X-DCM, as well as developing a program package to realize format interchange among DICOM, XML-DCM, and X-DCM. XML-DCM is based on the DICOM structure while replacing numeric tags with accessible XML character string tags. The merits are as following: a) every character string tag of XML-DCM has explicit meaning, so users can understand standard data elements and those private data elements easily without looking up the Data Dictionary. In this way, the readability and data sharing of DICOM files are greatly improved; b) According to requirements, users can set new character string tags with explicit meaning to their own system to extend the capacity of data elements; c) User can read the medical image and associated information conveniently through IE, ultimately enlarging the scope of data sharing. The application of storage format X-DCM will reduce data redundancy and save storage memory. The result of practical application shows that XML-DCM does favor integration and share of medical image data among different systems or devices.

  8. Extracting tag hierarchies.

    PubMed

    Tibély, Gergely; Pollner, Péter; Vicsek, Tamás; Palla, Gergely

    2013-01-01

    Tagging items with descriptive annotations or keywords is a very natural way to compress and highlight information about the properties of the given entity. Over the years several methods have been proposed for extracting a hierarchy between the tags for systems with a "flat", egalitarian organization of the tags, which is very common when the tags correspond to free words given by numerous independent people. Here we present a complete framework for automated tag hierarchy extraction based on tag occurrence statistics. Along with proposing new algorithms, we are also introducing different quality measures enabling the detailed comparison of competing approaches from different aspects. Furthermore, we set up a synthetic, computer generated benchmark providing a versatile tool for testing, with a couple of tunable parameters capable of generating a wide range of test beds. Beside the computer generated input we also use real data in our studies, including a biological example with a pre-defined hierarchy between the tags. The encouraging similarity between the pre-defined and reconstructed hierarchy, as well as the seemingly meaningful hierarchies obtained for other real systems indicate that tag hierarchy extraction is a very promising direction for further research with a great potential for practical applications. Tags have become very prevalent nowadays in various online platforms ranging from blogs through scientific publications to protein databases. Furthermore, tagging systems dedicated for voluntary tagging of photos, films, books, etc. with free words are also becoming popular. The emerging large collections of tags associated with different objects are often referred to as folksonomies, highlighting their collaborative origin and the "flat" organization of the tags opposed to traditional hierarchical categorization. Adding a tag hierarchy corresponding to a given folksonomy can very effectively help narrowing or broadening the scope of search. Moreover, recommendation systems could also benefit from a tag hierarchy.

  9. Extracting Tag Hierarchies

    PubMed Central

    Tibély, Gergely; Pollner, Péter; Vicsek, Tamás; Palla, Gergely

    2013-01-01

    Tagging items with descriptive annotations or keywords is a very natural way to compress and highlight information about the properties of the given entity. Over the years several methods have been proposed for extracting a hierarchy between the tags for systems with a "flat", egalitarian organization of the tags, which is very common when the tags correspond to free words given by numerous independent people. Here we present a complete framework for automated tag hierarchy extraction based on tag occurrence statistics. Along with proposing new algorithms, we are also introducing different quality measures enabling the detailed comparison of competing approaches from different aspects. Furthermore, we set up a synthetic, computer generated benchmark providing a versatile tool for testing, with a couple of tunable parameters capable of generating a wide range of test beds. Beside the computer generated input we also use real data in our studies, including a biological example with a pre-defined hierarchy between the tags. The encouraging similarity between the pre-defined and reconstructed hierarchy, as well as the seemingly meaningful hierarchies obtained for other real systems indicate that tag hierarchy extraction is a very promising direction for further research with a great potential for practical applications. Tags have become very prevalent nowadays in various online platforms ranging from blogs through scientific publications to protein databases. Furthermore, tagging systems dedicated for voluntary tagging of photos, films, books, etc. with free words are also becoming popular. The emerging large collections of tags associated with different objects are often referred to as folksonomies, highlighting their collaborative origin and the “flat” organization of the tags opposed to traditional hierarchical categorization. Adding a tag hierarchy corresponding to a given folksonomy can very effectively help narrowing or broadening the scope of search. Moreover, recommendation systems could also benefit from a tag hierarchy. PMID:24391901

  10. Differentiation of tumor from viable myocardium using cardiac tagging with MR imaging.

    PubMed

    Bouton, S; Yang, A; McCrindle, B W; Kidd, L; McVeigh, E R; Zerhouni, E A

    1991-01-01

    We report the application of myocardial tagging by MR to define tissue planes and differentiate contractile from noncontractile tissue in a neonate with congenital cardiac rhabdomyoma. Using custom-written pulse programming software, six 2 mm thick radiofrequency (RF) slice-selective presaturation pulses (tags) were used to label the chest wall and myocardium in a star pattern in diastole, approximately 60 ms before the R-wave gating trigger. This method successfully delineated the myocardium from noncontractile tumor, providing information that influenced clinical management. This RF tagging technique allowed us to confirm the exact intramyocardial location of a congenital cardiac tumor.

  11. Tagging Water Sources in Atmospheric Models

    NASA Technical Reports Server (NTRS)

    Bosilovich, M.

    2003-01-01

    Tagging of water sources in atmospheric models allows for quantitative diagnostics of how water is transported from its source region to its sink region. In this presentation, we review how this methodology is applied to global atmospheric models. We will present several applications of the methodology. In one example, the regional sources of water for the North American Monsoon system are evaluated by tagging the surface evaporation. In another example, the tagged water is used to quantify the global water cycling rate and residence time. We will also discuss the need for more research and the importance of these diagnostics in water cycle studies.

  12. Quantum tagging for tags containing secret classical data

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kent, Adrian

    Various authors have considered schemes for quantum tagging, that is, authenticating the classical location of a classical tagging device by sending and receiving quantum signals from suitably located distant sites, in an environment controlled by an adversary whose quantum information processing and transmitting power is potentially unbounded. All of the schemes proposed elsewhere in the literature assume that the adversary is able to inspect the interior of the tagging device. All of these schemes have been shown to be breakable if the adversary has unbounded predistributed entanglement. We consider here the case in which the tagging device contains a finitemore » key string shared with distant sites but kept secret from the adversary, and show this allows the location of the tagging device to be authenticated securely and indefinitely. Our protocol relies on quantum key distribution between the tagging device and at least one distant site, and demonstrates a new practical application of quantum key distribution. It also illustrates that the attainable security in position-based cryptography can depend crucially on apparently subtle details in the security scenario considered.« less

  13. Measuring and Predicting Tag Importance for Image Retrieval.

    PubMed

    Li, Shangwen; Purushotham, Sanjay; Chen, Chen; Ren, Yuzhuo; Kuo, C-C Jay

    2017-12-01

    Textual data such as tags, sentence descriptions are combined with visual cues to reduce the semantic gap for image retrieval applications in today's Multimodal Image Retrieval (MIR) systems. However, all tags are treated as equally important in these systems, which may result in misalignment between visual and textual modalities during MIR training. This will further lead to degenerated retrieval performance at query time. To address this issue, we investigate the problem of tag importance prediction, where the goal is to automatically predict the tag importance and use it in image retrieval. To achieve this, we first propose a method to measure the relative importance of object and scene tags from image sentence descriptions. Using this as the ground truth, we present a tag importance prediction model to jointly exploit visual, semantic and context cues. The Structural Support Vector Machine (SSVM) formulation is adopted to ensure efficient training of the prediction model. Then, the Canonical Correlation Analysis (CCA) is employed to learn the relation between the image visual feature and tag importance to obtain robust retrieval performance. Experimental results on three real-world datasets show a significant performance improvement of the proposed MIR with Tag Importance Prediction (MIR/TIP) system over other MIR systems.

  14. Effects of beta-phenylethylamine on dopaminergic neurons of the ventral tegmental area in the rat: a combined electrophysiological and microdialysis study.

    PubMed

    Ishida, Kota; Murata, Mikio; Katagiri, Nobuyuki; Ishikawa, Masago; Abe, Kenji; Kato, Masatoshi; Utsunomiya, Iku; Taguchi, Kyoji

    2005-08-01

    The effects of systemic administration of beta-phenylethylamine (beta-PEA) and microiontophoretically applied beta-PEA on the spontaneous discharge of dopamine (DA) neurons in the ventral tegmental area (VTA) of the anesthetized rat were examined. Intravenous administration of beta-PEA (1.0, 2.5, and 5.0 mg/kg) and microiontophoretic applications of beta-PEA caused inhibitory responses in DA neurons. Systemic administration and microiontophoretic applications of beta-PEA induced dose- or current-dependent responses. The systemic beta-PEA-induced inhibitory responses were reversed by pretreatment with the DA D(2) receptor antagonists haloperidol (0.5 mg/kg i.p.) and sulpiride (10 mg/kg i.p). Pretreatment with reserpine (5 mg/kg i.p. 24 h earlier) did not completely block the systemic administration of beta-PEA (2.5 mg/kg) inhibition. A microdialysis study of freely moving rats demonstrated that the extracellular DA level increased significantly in response to local application of beta-PEA (100 muM) in the VTA via a microdialysis probe, and local application of beta-PEA-stimulated somatodendritic DA release in the VTA. The beta-PEA-induced release of DA was calcium ion-independent and was enhanced by pretreatment with pertussis toxin. These findings indicate that beta-phenylethylamine inhibits DA neuron activity via DA D(2) autoreceptors in the rat VTA and that this inhibitory effect is mediated by the somatodendritic DA release.

  15. The Use of Affinity Tags to Overcome Obstacles in Recombinant Protein Expression and Purification.

    PubMed

    Amarasinghe, Chinthaka; Jin, Jian-Ping

    2015-01-01

    Research and industrial demands for recombinant proteins continue to increase over time for their broad applications in structural and functional studies and as therapeutic agents. These applications often require large quantities of recombinant protein at desirable purity, which highlights the importance of developing and improving production approaches that provide high level expression and readily achievable purity of recombinant protein. E. coli is the most widely used host for the expression of a diverse range of proteins at low cost. However, there are common pitfalls that can severely limit the expression of exogenous proteins, such as stability, low solubility and toxicity to the host cell. To overcome these obstacles, one strategy that has found to be promising is the use of affinity tags or carrier peptide to aid in the folding of the target protein, increase solubility, lower toxicity and increase the level of expression. In the meantime, the tags and fusion proteins can be designed to facilitate affinity purification. Since the fusion protein may not exhibit the native conformation of the target protein, various strategies have been developed to remove the tag during or after purification to avoid potential complications in structural and functional studies and to obtain native biological activities. Despite extensive research and rapid development along these lines, there are unsolved problems and imperfect applications. This focused review compares and contrasts various strategies that employ affinity tags to improve bacterial expression and to facilitate purification of recombinant proteins. The pros and cons of the approaches are discussed for more effective applications and new directions of future improvement.

  16. Fully Integrated Passive UHF RFID Tag for Hash-Based Mutual Authentication Protocol.

    PubMed

    Mikami, Shugo; Watanabe, Dai; Li, Yang; Sakiyama, Kazuo

    2015-01-01

    Passive radio-frequency identification (RFID) tag has been used in many applications. While the RFID market is expected to grow, concerns about security and privacy of the RFID tag should be overcome for the future use. To overcome these issues, privacy-preserving authentication protocols based on cryptographic algorithms have been designed. However, to the best of our knowledge, evaluation of the whole tag, which includes an antenna, an analog front end, and a digital processing block, that runs authentication protocols has not been studied. In this paper, we present an implementation and evaluation of a fully integrated passive UHF RFID tag that runs a privacy-preserving mutual authentication protocol based on a hash function. We design a single chip including the analog front end and the digital processing block. We select a lightweight hash function supporting 80-bit security strength and a standard hash function supporting 128-bit security strength. We show that when the lightweight hash function is used, the tag completes the protocol with a reader-tag distance of 10 cm. Similarly, when the standard hash function is used, the tag completes the protocol with the distance of 8.5 cm. We discuss the impact of the peak power consumption of the tag on the distance of the tag due to the hash function.

  17. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Swansiger, W.A.; Shepodd, T.J.; Phillips, M.L.F.

    The ability to identify the manufacturers and distributors of chemicals seized in raids of illicit drug labs would be of great value in controlling the diversion of these chemicals. We developed a tagging scheme based on the addition of sub-ppM concentrations of various combinations of rare-earth elements to the target chemicals and evaluated a number of techniques for detecting the tags. We developed soluble tags for tagging liquids and selected Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) as the preferred detection technique. We developed insoluble tags for tagging solids and developed methods to analyze them and mix them into solid precursors. Wemore » have successfully demonstrated the tagging of several solvents and two of the precursor chemicals used in one of the most popular clandestine methamphetamine syntheses (ephedrine reacting with hydriodic acid/red phosphorus). The tagging scheme is capable of yielding tens of thousands of signatures (using holmium as an internal standard and up to 9 rare-earths at up to 3 concentrations yields 3{sup 9} {minus} 1 = 19,682 signatures) and is applicable to most of the chemicals on the precursor and essential chemicals list. In the concentrations employed, the tags are safe enough to be added to pharmaceuticals and cheap enough to tag tanker loads of chemicals.« less

  18. Monitoring and localization of buried plastic natural gas pipes using passive RF tags

    NASA Astrophysics Data System (ADS)

    Mondal, Saikat; Kumar, Deepak; Ghazali, Mohd. Ifwat; Chahal, Prem; Udpa, Lalita; Deng, Yiming

    2018-04-01

    A passive harmonic radio frequency (RF) tag on the pipe with added sensing capabilities is proposed in this paper. Radio frequency identification (RFID) based tagging has already emerged as a potential solution for chemical sensing, location detection, animal tagging, etc. Harmonic transponders are already quite popular compared to conventional RFIDs due to their improved signal to noise ratio (SNR). However, the operating frequency, transmitted power and tag efficiency become critical issues for underground RFIDs. In this paper, a comprehensive on-tag sensing, power budget and frequency analyses is performed for buried harmonic tag design. Accurate tracking of infrastructure burial depth is proposed to reduce the probability of failure of underground pipelines. Burial depth is estimated using phase information of received signals at different frequencies calculated using genetic algorithm (GA) based optimization for post processing. Suitable frequency range is determined for a variety of soil with different moisture content for small tag-antenna size. Different types of harmonic tags such as 1) Schottky diode, 2) Non-linear Transmission Line (NLTL) were compared for underground applications. In this study, the power, frequency and tag design have been optimized to achieve small antenna size, minimum signal loss and simple reader circuit for underground detection at up to 5 feet depth in different soil medium and moisture contents.

  19. Web servicing the biological office.

    PubMed

    Szugat, Martin; Güttler, Daniel; Fundel, Katrin; Sohler, Florian; Zimmer, Ralf

    2005-09-01

    Biologists routinely use Microsoft Office applications for standard analysis tasks. Despite ubiquitous internet resources, information needed for everyday work is often not directly and seamlessly available. Here we describe a very simple and easily extendable mechanism using Web Services to enrich standard MS Office applications with internet resources. We demonstrate its capabilities by providing a Web-based thesaurus for biological objects, which maps names to database identifiers and vice versa via an appropriate synonym list. The client application ProTag makes these features available in MS Office applications using Smart Tags and Add-Ins. http://services.bio.ifi.lmu.de/prothesaurus/

  20. The first genome-level transcriptome of the wood-degrading fungus Phanerochaete chrysosporium grown on red oak.

    PubMed

    Sato, Shin; Feltus, F Alex; Iyer, Prashanti; Tien, Ming

    2009-06-01

    As part of an effort to determine all the gene products involved in wood degradation, we have performed massively parallel pyrosequencing on an expression library from the white rot fungus Phanerochaete chrysosporium grown in shallow stationary cultures with red oak as the carbon source. Approximately 48,000 high quality sequence tags (246 bp average length) were generated. 53% of the sequence tags aligned to 4,262 P. chrysosporium gene models, and an additional 18.5% of the tags reliably aligned to the P. chrysosporium genome providing evidence for 961 putative novel fragmented gene models. Due to their role in lignocellulose degradation, the secreted proteins were focused upon. Our results show that the four enzymes required for cellulose degradation: endocellulase, exocellulase CBHI, exocellulase CBHII, and beta-glucosidase are all produced. For hemicellulose degradation, not all known enzymes were produced, but endoxylanases, acetyl xylan esterases and mannosidases were detected. For lignin degradation, the role of peroxidases has been questioned; however, our results show that lignin peroxidase is highly expressed along with the H(2)O(2) generating enzyme, alcohol oxidase. The transcriptome snapshot reveals that H(2)O(2) generation and utilization are central in wood degradation. Our results also reveal new transcripts that encode extracellular proteins with no known function.

  1. N-terminal processing of affinity-tagged recombinant proteins purified by IMAC procedures.

    PubMed

    Mooney, Jane T; Fredericks, Dale P; Christensen, Thorkild; Bruun Schiødt, Christine; Hearn, Milton T W

    2015-07-01

    The ability of a new class of metal binding tags to facilitate the purification of recombinant proteins, exemplified by the tagged glutathione S-transferase and human growth hormone, from Escherichia coli fermentation broths and lysates has been further investigated. These histidine-containing tags exhibit high affinity for borderline metal ions chelated to the immobilised ligand, 1,4,7-triazacyclononane (tacn). The use of this tag-tacn immobilised metal ion affinity chromatography (IMAC) system engenders high selectivity with regard to host cell protein removal and permits facile tag removal from the E. coli-expressed recombinant protein. In particular, these tags were specifically designed to enable their efficient removal by the dipeptidyl aminopeptidase 1 (DAP-1), thus capturing the advantages of high substrate specificity and rates of cleavage. MALDI-TOF MS analysis of the cleaved products from the DAP-1 digestion of the recombinant N-terminally tagged proteins confirmed the complete removal of the tag within 4-12 h under mild experimental conditions. Overall, this study demonstrates that the use of tags specifically designed to target tacn-based IMAC resins offers a comprehensive and flexible approach for the purification of E. coli-expressed recombinant proteins, where complete removal of the tag is an essential prerequisite for subsequent application of the purified native proteins in studies aimed at delineating the molecular and cellular basis of specific biological processes. Copyright © 2015 John Wiley & Sons, Ltd.

  2. Sphingomonas paucimobilis beta-glucosidase Bgl1: a member of a new bacterial subfamily in glycoside hydrolase family 1.

    PubMed Central

    Marques, Ana Rita; Coutinho, Pedro M; Videira, Paula; Fialho, Arsénio M; Sá-Correia, Isabel

    2003-01-01

    The Sphingomonas paucimobilis beta-glucosidase Bgl1 is encoded by the bgl1 gene, associated with an 1308 bp open reading frame. The deduced protein has a potential signal peptide of 24 amino acids in the N-terminal region, and experimental evidence is consistent with the processing and export of the Bgl1 protein through the inner membrane to the periplasmic space. A His(6)-tagged 44.3 kDa protein was over-produced in the cytosol of Escherichia coli from a recombinant plasmid, which contained the S. paucimobilis bgl1 gene lacking the region encoding the putative signal peptide. Mature beta-glucosidase Bgl1 is specific for aryl-beta-glucosides and has no apparent activity with oligosaccharides derived from cellulose hydrolysis and other saccharides. A structure-based alignment established structural relations between S. paucimobilis Bgl1 and other members of the glycoside hydrolase (GH) family 1 enzymes. At subsite -1, the conserved residues required for catalysis by GH1 enzymes are present in Bgl1 with only minor differences. Major differences are found at subsite +1, the aglycone binding site. This alignment seeded a sequence-based phylogenetic analysis of GH1 enzymes, revealing an absence of horizontal transfer between phyla. Bootstrap analysis supported the definition of subfamilies and revealed that Bgl1, the first characterized beta-glucosidase from the genus Sphingomonas, represents a very divergent bacterial subfamily, closer to archaeal subfamilies than to others of bacterial origin. PMID:12444924

  3. Hemoglobin Abraham Lincoln, beta32 (B14) leucine leads to proline. An unstable variant producing severe hemolytic disease.

    PubMed

    Honig, G R; Green, D; Shamsuddin, M; Vida, L N; Mason, R G; Gnarra, D J; Maurer, H S

    1973-07-01

    An unstable hemoglobin variant was identified in a Negro woman with hemolytic anemia since infancy. A splenectomy had been performed when the patient was a child. The anemia was accompanied by erythrocyte inclusion bodies and excretion of darkly pigmented urine. Neither parent of the proposita demonstrated any hematologic abnormality, and it appeared that this hemoglobin variant arose as a new mutation. Erythrocyte survival in the patient was greatly reduced: the erythrocyte t(1/2) using radiochromium as a tag was 2.4 days, and a reticulocyte survival study performed after labeling the cells with L-[(14)C]leucine indicated a t(1/2) of 7.2 days. When stroma-free hemolysates were heated at 50 degrees C, 16-20% of the hemoglobin precipitated. The thermolability was prevented by the addition of hemin, carbon monoxide, or dithionite, suggesting an abnormality of heme binding. An increased rate of methemoglobin formation was also observed after incubation of erythrocytes at 37 degrees C. The abnormal hemoglobin could not be separated from hemoglobin A by electrophoresis or chromatography, but it was possible to isolate the variant beta-chain by precipitation with p-hydroxymercuribenzoate. Purification of the beta-chain by column chromatography followed by peptide mapping and amino acid analysis demonstrated a substitution of proline for beta32 leucine. It appears likely that a major effect of this substitution is a disruption of the normal orientation of the adjacent leucine residue at beta31 to impair heme stabilization.

  4. Engineering protein scaffolds for protein separation, biocatalysis and nanotechnology applications

    NASA Astrophysics Data System (ADS)

    Liu, Fang

    Globally, there is growing appreciation for developing a sustainable economy that uses eco-efficient bio-processes. Biotechnology provides an increasing range of tools for industry to help reduce cost and improve environmental performance. Inspired by the naturally evolved machineries of protein scaffolds and their binding ligands, synthetic protein scaffolds were engineered based on cohesin-dockerin interactions and metal chelating peptides to tackle the challenges and make improvements in three specific areas: (1) protein purification, (2) biofuel cells, and (3) nanomaterial synthesis. The first objective was to develop efficient and cost-effective non-chromatographic purification processes to purify recombinant proteins in an effort to meet the dramatically growing market of protein drugs. In our design, the target protein was genetically fused with a dockerin domain from Clostridium thermocellum and direct purification and recovery was achieved using thermo-responsive elastin-like polypeptide (ELP) scaffold containing the cohesin domain from the same species. By exploiting the highly specific interaction between the dockerin and cohesin domain and the reversible aggregation property of ELP, highly purified and active dockerin-tagged proteins, such as endoglucanase CelA, chloramphenicol acetyl transferase (CAT) and enhanced green fluorescence protein (EGFP), were recovered directly from crude cell extracts in a single purification step with yields achieving over 90%. Incorporation of a self-cleaving intein domain enabled rapid removal of the affinity tag from the target proteins by another cycle of thermal precipitation. The purification cost can be further reduced by regenerating and recycling the ELP-cohesin capturing scaffolds. However, due to the high binding affinity between cohesin and dockerin domains, the bound dockerin-intein tag cannot be completely disassociated from ELP-cohesin scaffold after binding. Therefore, a truncated dockerin with the calcium-coordinating function impaired was used in replace of the original full length dockerin domain. The truncated dockerin domain maintained its functionality as an effective affinity tag, and efficient EDTA mediated dissociation of the bound dockerin-intein tag was also realized. The regenerated ELP capturing scaffold was reused for additional purification cycles without any decrease in efficiency. The second objective was to assemble biocatalysts for biofuel cells. Three beta-NAD dependent dehydrogenases, alcohol dehydrogenase (ADH), formaldehyde dehydrogenase (FALDH) and formate dehydrogenase (FDH), were site-specifically co-localized onto the scaffolds displayed on the yeast surface based on the high-affinity interactions between three orthogonal cohesin/dockerin pairs. The assembled multi-enzyme cascades, which can completely convert methanol to CO2, showed improved production yield compared with that of the non-complexed enzyme mixture, indicating efficient substrate channeling among the three enzymes. This strategy can be easily extended to other complex cascade reactions for enzymatic fuel cell applications. To further explore the role of biotechnology toward environmental sustainability, Escherichia coli was engineered to express phytochelatin synthase, which converted glutathione into the metal-binding peptide phytochelatin (PC). PCs served as peptide scaffolds and mediated synthesis of CdS nanocrystals. This approach may be generalized to guide the in vitro self-assembly of a wide range of nanocrystals with different compositions and sizes.

  5. Long range ultra-high frequency (UHF) radio frequency identification (RFID) antenna design

    NASA Astrophysics Data System (ADS)

    Reynolds, Nathan D.

    There is an ever-increasing demand for radio frequency identification (RFID) tags that are passive, long range, and mountable on multiple surfaces. Currently, RFID technology is utilized in numerous applications such as supply chain management, access control, and public transportation. With the combination of sensory systems in recent years, the applications of RFID technology have been extended beyond tracking and identifying. This extension includes applications such as environmental monitoring and healthcare applications. The available sensory systems usually operate in the medium or high frequency bands and have a low read range. However, the range limitations of these systems are being overcome by the development of RFID sensors focused on utilizing tags in the ultra-high frequency (UHF) band. Generally, RFID tags have to be mounted to the object that is being identified. Often the objects requiring identification are metallic. The inherent properties of metallic objects have substantial effects on nearby electromagnetic radiation; therefore, the operation of the tag antenna is affected when mounted on a metallic surface. This outlines one of the most challenging problems for RFID systems today: the optimization of tag antenna performance in a complex environment. In this research, a novel UHF RFID tag antenna, which has a low profile, long range, and is mountable on metallic surfaces, is designed analytically and simulated using a 3-D electromagnetic simulator, ANSYS HFSS. A microstrip patch antenna is selected as the antenna structure, as patch antennas are low profile and suitable for mounting on metallic surfaces. Matching and theoretical models of the microstrip patch antenna are investigated. Once matching and theory of a microstrip patch antenna is thoroughly understood, a unique design technique using electromagnetic band gap (EBG) structures is explored. This research shows that the utilization of an EBG structure in the patch antenna design yields an improvement in gain, or range, and in the ability to be mounted on multiple metallic surfaces.

  6. Colloid-based multiplexed method for screening plant biomass-degrading glycoside hydrolase activities in microbial communities

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Reindl, W.; Deng, K.; Gladden, J.M.

    2011-05-01

    The enzymatic hydrolysis of long-chain polysaccharides is a crucial step in the conversion of biomass to lignocellulosic biofuels. The identification and characterization of optimal glycoside hydrolases is dependent on enzyme activity assays, however existing methods are limited in terms of compatibility with a broad range of reaction conditions, sample complexity, and especially multiplexity. The method we present is a multiplexed approach based on Nanostructure-Initiator Mass Spectrometry (NIMS) that allowed studying several glycolytic activities in parallel under diverse assay conditions. Although the substrate analogs carried a highly hydrophobic perfluorinated tag, assays could be performed in aqueous solutions due colloid formation ofmore » the substrate molecules. We first validated our method by analyzing known {beta}-glucosidase and {beta}-xylosidase activities in single and parallel assay setups, followed by the identification and characterization of yet unknown glycoside hydrolase activities in microbial communities.« less

  7. RFID tags as a direct tracer for water and sediment dynamics

    NASA Astrophysics Data System (ADS)

    Sommerer, Erik; Plate, Simon; Güntner, Andreas

    2014-05-01

    RFID (Radio Frequency IDentification) is a wireless automatic identification system to track objects with widespread application in industrial operations, but also selected applications in ecological research (animal tracking) and for hydro-sedimentological studies (sediment transport with RFID tags embedded in bedload material). In this study, for the first time, we test and apply RFID tags as a direct tracer to track water pathways, erosion patterns and sediment transport on the surface at the hillslope and headwater scale. The RFID system used here consists of tags with a size of 12 x 2 mm and a combination of mobile and stationary antennas. The transport pathways and velocities of the RFID tags can be individually assessed due to their unique identification numbers. The study area is a badland of easily erodible marls and carbonates located in the Villacarli catchment (42 km²) in the Central Spanish Pyrenees. The badlands have been identified as one of the main sediment sources for siltation of the downstream Barasona Reservoir. More than 700 tags were placed in different terrain units using three experimental setups, including lab experiments: (i) intensive feasibility tests ranging from laboratory flume experiments to tracer studies under natural channel and slope conditions to compare the transport of RFID tags relative to colored particles of the natural sediment; (ii) several transects across the badland to investigate sediment transfer characteristics on different morphological units (i.e. channel, rills, slopes); (iii) a raster of 99 RFID tags covering a slope flank with vegetated and unvegetated parts to reveal the influence of vegetation to erosion and transport processes. The detection of transported tags was carried out with a mobile antenna system to map the spatial distribution of tags after selected rainfall events and with two stationary antennas in channel cross-sections for time-continuous observation of tag passage. From the observations, we derived transport distances and velocities for a one year period. The transport behavior of the RFID tags was similar to the micrite limestone material of the badland. These results indicate the important role of terrain position and vegetation patches for erosion patterns, sediment transfer and the associated time scales along the catchment. We assess the potentials and limitations of the RFID technique as a direct tracer for assessing water and sediment connectivity along hillslopes to first order channels.

  8. Live single-cell laser tag.

    PubMed

    Binan, Loïc; Mazzaferri, Javier; Choquet, Karine; Lorenzo, Louis-Etienne; Wang, Yu Chang; Affar, El Bachir; De Koninck, Yves; Ragoussis, Jiannis; Kleinman, Claudia L; Costantino, Santiago

    2016-05-20

    The ability to conduct image-based, non-invasive cell tagging, independent of genetic engineering, is key to cell biology applications. Here we introduce cell labelling via photobleaching (CLaP), a method that enables instant, specific tagging of individual cells based on a wide array of criteria such as shape, behaviour or positional information. CLaP uses laser illumination to crosslink biotin onto the plasma membrane, coupled with streptavidin conjugates to label individual cells for genomic, cell-tracking, flow cytometry or ultra-microscopy applications. We show that the incorporated mark is stable, non-toxic, retained for several days, and transferred by cell division but not to adjacent cells in culture. To demonstrate the potential of CLaP for genomic applications, we combine CLaP with microfluidics-based single-cell capture followed by transcriptome-wide next-generation sequencing. Finally, we show that CLaP can also be exploited for inducing transient cell adhesion to substrates for microengineering cultures with spatially patterned cell types.

  9. Metabolic engineering of biomass for high energy density: oilseed-like triacylglycerol yields from plant leaves.

    PubMed

    Vanhercke, Thomas; El Tahchy, Anna; Liu, Qing; Zhou, Xue-Rong; Shrestha, Pushkar; Divi, Uday K; Ral, Jean-Philippe; Mansour, Maged P; Nichols, Peter D; James, Christopher N; Horn, Patrick J; Chapman, Kent D; Beaudoin, Frederic; Ruiz-López, Noemi; Larkin, Philip J; de Feyter, Robert C; Singh, Surinder P; Petrie, James R

    2014-02-01

    High biomass crops have recently attracted significant attention as an alternative platform for the renewable production of high energy storage lipids such as triacylglycerol (TAG). While TAG typically accumulates in seeds as storage compounds fuelling subsequent germination, levels in vegetative tissues are generally low. Here, we report the accumulation of more than 15% TAG (17.7% total lipids) by dry weight in Nicotiana tabacum (tobacco) leaves by the co-expression of three genes involved in different aspects of TAG production without severely impacting plant development. These yields far exceed the levels found in wild-type leaf tissue as well as previously reported engineered TAG yields in vegetative tissues of Arabidopsis thaliana and N. tabacum. When translated to a high biomass crop, the current levels would translate to an oil yield per hectare that exceeds those of most cultivated oilseed crops. Confocal fluorescence microscopy and mass spectrometry imaging confirmed the accumulation of TAG within leaf mesophyll cells. In addition, we explored the applicability of several existing oil-processing methods using fresh leaf tissue. Our results demonstrate the technical feasibility of a vegetative plant oil production platform and provide for a step change in the bioenergy landscape, opening new prospects for sustainable food, high energy forage, biofuel and biomaterial applications. © 2013 CSIRO. Plant Biotechnology Journal published by Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  10. TaGS5-3A, a grain size gene selected during wheat improvement for larger kernel and yield.

    PubMed

    Ma, Lin; Li, Tian; Hao, Chenyang; Wang, Yuquan; Chen, Xinhong; Zhang, Xueyong

    2016-05-01

    Grain size is a dominant component of grain weight in cereals. Earlier studies have shown that OsGS5 plays a major role in regulating both grain size and weight in rice via promotion of cell division. In this study, we isolated TaGS5 homoeologues in wheat and mapped them on chromosomes 3A, 3B and 3D. Temporal and spatial expression analysis showed that TaGS5 homoeologues were preferentially expressed in young spikes and developing grains. Two alleles of TaGS5-3A, TaGS5-3A-T and TaGS5-3A-G were identified in wheat accessions, and a functional marker was developed to discriminate them. Association analysis revealed that TaGS5-3A-T was significantly correlated with larger grain size and higher thousand kernel weight. Biochemical assays showed that TaGS5-3A-T possesses a higher enzymatic activity than TaGS5-3A-G. Transgenic rice lines overexpressing TaGS5-3A-T also exhibited larger grain size and higher thousand kernel weight than TaGS5-3A-G lines, and the transcript levels of cell cycle-related genes in TaGS5-3A-T lines were higher than those in TaGS5-3A-G lines. Furthermore, systematic evolution analysis in diploid, tetraploid and hexaploid wheat showed that TaGS5-3A underwent strong artificial selection during wheat polyploidization events and the frequency changes of two alleles demonstrated that TaGS5-3A-T was favoured in global modern wheat cultivars. These results suggest that TaGS5-3A is a positive regulator of grain size and its favoured allele TaGS5-3A-T exhibits a larger potential application in wheat high-yield breeding. © 2015 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  11. p38 Mitogen-Activated Protein Kinase in Metastasis Associated with Transforming Growth Factor Beta

    DTIC Science & Technology

    2006-06-01

    Functional Analysis of cells • Expression of HA-tagged TβRI • Signaling (p-Smad2, p- HSP27 , p-p38) • Transcription (SBE-Lux, 3TP-Lux) • Proliferation...when they also affected lls expressing DN-p38alpha phosphorylation of HSP27 was effectively blocked (data not shown). However, cells e studies show...231-ALK5-T204D cells treated with kinase inhibitors U0126, SB202190, SP600125 (JNK). (B) Immunoblot of phospho-Smad2/3, phospho- HSP27 , and alpha

  12. Wnt/Beta-Catenin, Foxa2, and CXCR4 Axis Controls Prostate Cancer Progression

    DTIC Science & Technology

    2013-07-01

    NT1 cells with or without Foxa2 were cultured in 5% strip medium in the presence or absence of androgen. Cell growth curve was measured by WTS...assays. Over-expression of Foxa2 increased NT1 cell growth even in the absence of androgen. Figure 3. knocking-down Foxa2 did not affect PC3 cell ...in NeoTag1 cells decreased AR level; however, AR 6 activity did not change. Foxa2 target genes were up-regulated in NT1 /Foxa2 over-expressing

  13. B(s) Mixing, Delta Gamma(s) and CP Violation at the Tevatron

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Di Giovanni, Gian Piero; /Paris U., VI-VII

    The authors discuss the results from the Tevatron experiments on mixing and CP violation in the B{sub s}{sup 0}-{bar B}{sub s}{sup 0} system, with particular emphasis to the updated measurements of the decay-width difference {Delta}{Lambda}{sub s} and the first measurement of the CP-violating phase {beta}{sub s} using flavor tagging information. They also briefly review the charge asymmetry measurements in semileptonic B{sub s}{sup 0} decays and in B{sup {+-}} {yields} J/{psi}K{sup {+-}} decays.

  14. Weld failure detection

    DOEpatents

    Pennell, William E.; Sutton, Jr., Harry G.

    1981-01-01

    Method and apparatus for detecting failure in a welded connection, particrly applicable to not readily accessible welds such as those joining components within the reactor vessel of a nuclear reactor system. A preselected tag gas is sealed within a chamber which extends through selected portions of the base metal and weld deposit. In the event of a failure, such as development of a crack extending from the chamber to an outer surface, the tag gas is released. The environment about the welded area is directed to an analyzer which, in the event of presence of the tag gas, evidences the failure. A trigger gas can be included with the tag gas to actuate the analyzer.

  15. Application of the Terrestar Satellite Constellation to the Global Initiative for Tracking Special and Nonproliferation Material

    DTIC Science & Technology

    2011-09-01

    A.  EXPERIMENT BACKGROUND ................................................................35  1.  BlackBerry Messenger...in Orbit (From [Terrestar, 2011])..................................................24  Figure 11.  BlackBerry Tag...36  Figure 12.  OV-1 Diagram of BlackBerry Tag

  16. A CMOS Pressure Sensor Tag Chip for Passive Wireless Applications

    PubMed Central

    Deng, Fangming; He, Yigang; Li, Bing; Zuo, Lei; Wu, Xiang; Fu, Zhihui

    2015-01-01

    This paper presents a novel monolithic pressure sensor tag for passive wireless applications. The proposed pressure sensor tag is based on an ultra-high frequency RFID system. The pressure sensor element is implemented in the 0.18 µm CMOS process and the membrane gap is formed by sacrificial layer release, resulting in a sensitivity of 1.2 fF/kPa within the range from 0 to 600 kPa. A three-stage rectifier adopts a chain of auxiliary floating rectifier cells to boost the gate voltage of the switching transistors, resulting in a power conversion efficiency of 53% at the low input power of −20 dBm. The capacitive sensor interface, using phase-locked loop archietcture, employs fully-digital blocks, which results in a 7.4 bits resolution and 0.8 µW power dissipation at 0.8 V supply voltage. The proposed passive wireless pressure sensor tag costs a total 3.2 µW power dissipation. PMID:25806868

  17. A CMOS pressure sensor tag chip for passive wireless applications.

    PubMed

    Deng, Fangming; He, Yigang; Li, Bing; Zuo, Lei; Wu, Xiang; Fu, Zhihui

    2015-03-23

    This paper presents a novel monolithic pressure sensor tag for passive wireless applications. The proposed pressure sensor tag is based on an ultra-high frequency RFID system. The pressure sensor element is implemented in the 0.18 µm CMOS process and the membrane gap is formed by sacrificial layer release, resulting in a sensitivity of 1.2 fF/kPa within the range from 0 to 600 kPa. A three-stage rectifier adopts a chain of auxiliary floating rectifier cells to boost the gate voltage of the switching transistors, resulting in a power conversion efficiency of 53% at the low input power of -20 dBm. The capacitive sensor interface, using phase-locked loop archietcture, employs fully-digital blocks, which results in a 7.4 bits resolution and 0.8 µW power dissipation at 0.8 V supply voltage. The proposed passive wireless pressure sensor tag costs a total 3.2 µW power dissipation.

  18. Completely monodisperse, highly repetitive proteins for bioconjugate capillary electrophoresis: Development and characterization

    PubMed Central

    Lin, Jennifer S.; Albrecht, Jennifer Coyne; Meagher, Robert J.; Wang, Xiaoxiao; Barron, Annelise E.

    2011-01-01

    Protein-based polymers are increasingly being used in biomaterial applications due to their ease of customization and potential monodispersity. These advantages make protein polymers excellent candidates for bioanalytical applications. Here we describe improved methods for producing drag-tags for Free-Solution Conjugate Electrophoresis (FSCE). FSCE utilizes a pure, monodisperse recombinant protein, tethered end-on to a ssDNA molecule, to enable DNA size separation in aqueous buffer. FSCE also provides a highly sensitive method to evaluate the polydispersity of a protein drag-tag and thus its suitability for bioanalytical uses. This method is able to detect slight differences in drag-tag charge or mass. We have devised an improved cloning, expression, and purification strategy that enables us to generate, for the first time, a truly monodisperse 20 kDa protein polymer and a nearly monodisperse 38 kDa protein. These newly produced proteins can be used as drag-tags to enable longer read DNA sequencing by free-solution microchannel electrophoresis. PMID:21553840

  19. Rapid Analysis of Microalgal Triacylglycerols with Direct-Infusion Mass Spectrometry

    DOE PAGES

    Christensen, Earl; Sudasinghe, Nilusha; Dandamudi, Kodanda Phani Raj; ...

    2015-09-01

    Cultivation of microalgae has the potential to provide lipid-derived feedstocks for conversion to liquid transportation fuels. Lipid extracts from microalgae are significantly more complex than those of traditional seed oils, and their composition changes significantly throughout the microalgal growth period. With three acyl side chains per molecule, triglycerides (TAGs) are an important fuel precursor, and the distribution of acyl chain composition for TAGs has a significant impact on fuel properties and processing. Therefore, determination of the distribution of microalgal TAG production is needed to assess the value of algal extracts designed for fuel production and to optimize strain, cultivation, andmore » harvesting practices. Methods utilized for TAG speciation commonly involve complicated and time-consuming chromatographic techniques. Here we present a method for TAG speciation and quantification based on direct-infusion mass spectrometry, which provides rapid characterization of TAG profiles without chromatographic separation. Specifically, we utilize Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) to provide a reference library of TAGs for the microalgae Nannochloropsis sp. that provides the basis for high-throughput TAG quantitation by time-of-flight mass spectrometry (TOF MS). In conclusion, we demonstrate the application of this novel approach for lipid characterization with respect to TAG compound distribution, which informs both immediate and future strain and process optimization strategies.« less

  20. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Christensen, Earl; Sudasinghe, Nilusha; Dandamudi, Kodanda Phani Raj

    Cultivation of microalgae has the potential to provide lipid-derived feedstocks for conversion to liquid transportation fuels. Lipid extracts from microalgae are significantly more complex than those of traditional seed oils, and their composition changes significantly throughout the microalgal growth period. With three acyl side chains per molecule, triglycerides (TAGs) are an important fuel precursor, and the distribution of acyl chain composition for TAGs has a significant impact on fuel properties and processing. Therefore, determination of the distribution of microalgal TAG production is needed to assess the value of algal extracts designed for fuel production and to optimize strain, cultivation, andmore » harvesting practices. Methods utilized for TAG speciation commonly involve complicated and time-consuming chromatographic techniques. Here we present a method for TAG speciation and quantification based on direct-infusion mass spectrometry, which provides rapid characterization of TAG profiles without chromatographic separation. Specifically, we utilize Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) to provide a reference library of TAGs for the microalgae Nannochloropsis sp. that provides the basis for high-throughput TAG quantitation by time-of-flight mass spectrometry (TOF MS). In conclusion, we demonstrate the application of this novel approach for lipid characterization with respect to TAG compound distribution, which informs both immediate and future strain and process optimization strategies.« less

  1. Fully Integrated Passive UHF RFID Tag for Hash-Based Mutual Authentication Protocol

    PubMed Central

    Mikami, Shugo; Watanabe, Dai; Li, Yang; Sakiyama, Kazuo

    2015-01-01

    Passive radio-frequency identification (RFID) tag has been used in many applications. While the RFID market is expected to grow, concerns about security and privacy of the RFID tag should be overcome for the future use. To overcome these issues, privacy-preserving authentication protocols based on cryptographic algorithms have been designed. However, to the best of our knowledge, evaluation of the whole tag, which includes an antenna, an analog front end, and a digital processing block, that runs authentication protocols has not been studied. In this paper, we present an implementation and evaluation of a fully integrated passive UHF RFID tag that runs a privacy-preserving mutual authentication protocol based on a hash function. We design a single chip including the analog front end and the digital processing block. We select a lightweight hash function supporting 80-bit security strength and a standard hash function supporting 128-bit security strength. We show that when the lightweight hash function is used, the tag completes the protocol with a reader-tag distance of 10 cm. Similarly, when the standard hash function is used, the tag completes the protocol with the distance of 8.5 cm. We discuss the impact of the peak power consumption of the tag on the distance of the tag due to the hash function. PMID:26491714

  2. A binding energy study of the Atomic Mass Evaluation 2012 and an updated beta-decay study of neutron-rich 74Cu

    NASA Astrophysics Data System (ADS)

    Tracy, James L., Jr.

    A study of ground state binding energy values listed in the Atomic Mass Evaluation 2012 (AME2012) using an interpretive approach, as opposed to the exploratory methods of previous models, is presented. This model is based on a postulate requiring all protons to pair with available neutrons to form bound alpha clusters as the ground state for an N = Z core upon which excess neutrons are added. For each core, the trend of the binding energy as a function of excess neutrons in the isotopic chain can be fit with a three-term quadratic function. The quadratic parameter reveals a smooth decaying exponential function. By re-envisioning the determination of mass excess, the constant-term fit parameters, representing N = Z nuclei, reveal a near-symmetry around Z = 50. The linear fit parameters exhibit trends which are linear functions of core size. A neutron drip-line prediction is compared against current models. By considering the possibility of an alpha-cluster core, a new ground-state structure grouping scheme is presented; nucleon-nucleon pairing is shown to have a greater role in level filling. This model, referred to as the Alpha-Deuteron-Neutron Model, yields promising first results when considering root-mean-square variances from the AME2012. The beta-decay of the neutron-rich isotope 74Cu has been studied using three high-purity Germanium clover detectors at the Holifield Radioactive Ion Beam Facility at Oak Ridge National Laboratory. A high-resolution mass separator greatly improved the purity of the 74Cu beam by removing isobaric contaminants, thus allowing decay through its isobar chain to the stable 74Ge at the center of the LeRIBSS detector array without any decay chain member dominating. Using coincidence gating techniques, 121 gamma-rays associated with 74Cu were isolated from the collective singles spectrum. Eighty-seven of these were placed in an expanded level scheme, and updated beta-feeding level intensities and log( ft) values are presented based on multiple newly-placed excited states up to 6.8 MeV. The progression of simulated Total Absorption gamma-ray Spectroscopy (TAGS) based on known levels and beta feeding values from previous measurements to this evaluation are presented and demonstrate the need for a TAGS measurement of this isotope to gain a more complete understanding of its decay scheme.

  3. Passive RFID Rotation Dimension Reduction via Aggregation

    NASA Astrophysics Data System (ADS)

    Matthews, Eric

    Radio Frequency IDentification (RFID) has applications in object identification, position, and orientation tracking. RFID technology can be applied in hospitals for patient and equipment tracking, stores and warehouses for product tracking, robots for self-localisation, tracking hazardous materials, or locating any other desired object. Efficient and accurate algorithms that perform localisation are required to extract meaningful data beyond simple identification. A Received Signal Strength Indicator (RSSI) is the strength of a received radio frequency signal used to localise passive and active RFID tags. Many factors affect RSSI such as reflections, tag rotation in 3D space, and obstacles blocking line-of-sight. LANDMARC is a statistical method for estimating tag location based on a target tag's similarity to surrounding reference tags. LANDMARC does not take into account the rotation of the target tag. By either aggregating multiple reference tag positions at various rotations, or by determining a rotation value for a newly read tag, we can perform an expected value calculation based on a comparison to the k-most similar training samples via an algorithm called K-Nearest Neighbours (KNN) more accurately. By choosing the average as the aggregation function, we improve the relative accuracy of single-rotation LANDMARC localisation by 10%, and any-rotation localisation by 20%.

  4. Development of a Dehalogenase-Based Protein Fusion Tag Capable of Rapid, Selective and Covalent Attachment to Customizable Ligands

    PubMed Central

    Encell, Lance P; Friedman Ohana, Rachel; Zimmerman, Kris; Otto, Paul; Vidugiris, Gediminas; Wood, Monika G; Los, Georgyi V; McDougall, Mark G; Zimprich, Chad; Karassina, Natasha; Learish, Randall D; Hurst, Robin; Hartnett, James; Wheeler, Sarah; Stecha, Pete; English, Jami; Zhao, Kate; Mendez, Jacqui; Benink, Hélène A; Murphy, Nancy; Daniels, Danette L; Slater, Michael R; Urh, Marjeta; Darzins, Aldis; Klaubert, Dieter H; Bulleit, Robert F; Wood, Keith V

    2012-01-01

    Our fundamental understanding of proteins and their biological significance has been enhanced by genetic fusion tags, as they provide a convenient method for introducing unique properties to proteins so that they can be examinedin isolation. Commonly used tags satisfy many of the requirements for applications relating to the detection and isolation of proteins from complex samples. However, their utility at low concentration becomes compromised if the binding affinity for a detection or capture reagent is not adequate to produce a stable interaction. Here, we describe HaloTag® (HT7), a genetic fusion tag based on a modified haloalkane dehalogenase designed and engineered to overcome the limitation of affinity tags by forming a high affinity, covalent attachment to a binding ligand. HT7 and its ligand have additional desirable features. The tag is relatively small, monomeric, and structurally compatible with fusion partners, while the ligand is specific, chemically simple, and amenable to modular synthetic design. Taken together, the design features and molecular evolution of HT7 have resulted in a superior alternative to common tags for the overexpression, detection, and isolation of target proteins. PMID:23248739

  5. Construction of a chimeric thermostable pyrophosphatase to facilitate its purification and immobilization by using the choline-binding tag.

    PubMed

    Moldes, Cristina; García, José L; García, Pedro

    2004-08-01

    The thermophilic inorganic pyrophosphatase (Pyr) from Thermus thermophilus has been produced in Escherichia coli fused to the C terminus of the choline-binding tag (ChB tag) derived from the choline-binding domain (ChBD) of pneumococcal LytA autolysin. The chimeric ChBD-Pyr protein retains its thermostable activity and can be purified in a single step by DEAE-cellulose affinity chromatography. Pyr can be further released from the ChBD by thrombin, using the specific protease recognition site incorporated in the C terminus of this tag. Remarkably, the ChB tag provides a selective and very strong thermostable noncovalent immobilization of ChBD-Pyr in the DEAE-cellulose matrix. The binding of choline or choline analogues, such as DEAE, confers a high thermal stability to this tag; therefore, the immobilized chimeric enzyme can be assayed at high temperature without protein leakage, demonstrating the usefulness of the ChB tag for noncovalent immobilization of thermophilic proteins. Moreover, ChBD-Pyr can be purified and immobilized in a single step on commercial DEAE-cellulose paper. The affinity of the ChB tag for this versatile solid support can be very helpful in developing many biotechnological applications.

  6. Evolving effective behaviours to interact with tag-based populations

    NASA Astrophysics Data System (ADS)

    Yucel, Osman; Crawford, Chad; Sen, Sandip

    2015-07-01

    Tags and other characteristics, externally perceptible features that are consistent among groups of animals or humans, can be used by others to determine appropriate response strategies in societies. This usage of tags can be extended to artificial environments, where agents can significantly reduce cognitive effort spent on appropriate strategy choice and behaviour selection by reusing strategies for interacting with new partners based on their tags. Strategy selection mechanisms developed based on this idea have successfully evolved stable cooperation in games such as the Prisoner's Dilemma game but relies upon payoff sharing and matching methods that limit the applicability of the tag framework. Our goal is to develop a general classification and behaviour selection approach based on the tag framework. We propose and evaluate alternative tag matching and adaptation schemes for a new, incoming individual to select appropriate behaviour against any population member of an existing, stable society. Our proposed approach allows agents to evolve both the optimal tag for the environment as well as appropriate strategies for existing agent groups. We show that these mechanisms will allow for robust selection of optimal strategies by agents entering a stable society and analyse the various environments where this approach is effective.

  7. 76 FR 44304 - Submission for OMB Review; Comment Request

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-25

    ... (tunas, sharks, swordfish, and billfish). Archival tags collect location, temperature, and water depth data that is useful for scientists researching the movements and behavior of individual fish. It is... tagged fish are exempted from other normally applicable regulations (i.e., immediate release of the fish...

  8. Exploring the Presence of microDNAs in Prostate Cancer Cell Lines, Tissue, and Sera of Prostate Cancer Patients and its Possible Application as Biomarker

    DTIC Science & Technology

    2016-04-01

    Sequence tags were mapped on the human reference genome using the Novoalign software. Only those...ends of the linear islands to create a novel junctional sequence that does not exist in the genome . Thus the PE- sequence of a fragment that breaks at... genome (Fig. 3b). Those PE-tags where one tag maps uniquely to an island and the other remains unmapped, but passes the sequence quality filter,

  9. RAC-tagging: Recombineering And Cas9-assisted targeting for protein tagging and conditional analyses

    PubMed Central

    Baker, Oliver; Gupta, Ashish; Obst, Mandy; Zhang, Youming; Anastassiadis, Konstantinos; Fu, Jun; Stewart, A. Francis

    2016-01-01

    A fluent method for gene targeting to establish protein tagged and ligand inducible conditional loss-of-function alleles is described. We couple new recombineering applications for one-step cloning of gRNA oligonucleotides and rapid generation of short-arm (~1 kb) targeting constructs with the power of Cas9-assisted targeting to establish protein tagged alleles in embryonic stem cells at high efficiency. RAC (Recombineering And Cas9)-tagging with Venus, BirM, APEX2 and the auxin degron is facilitated by a recombineering-ready plasmid series that permits the reuse of gene-specific reagents to insert different tags. Here we focus on protein tagging with the auxin degron because it is a ligand-regulated loss-of-function strategy that is rapid and reversible. Furthermore it includes the additional challenge of biallelic targeting. Despite high frequencies of monoallelic RAC-targeting, we found that simultaneous biallelic targeting benefits from long-arm (>4 kb) targeting constructs. Consequently an updated recombineering pipeline for fluent generation of long arm targeting constructs is also presented. PMID:27216209

  10. Clone tag detection in distributed RFID systems.

    PubMed

    Kamaludin, Hazalila; Mahdin, Hairulnizam; Abawajy, Jemal H

    2018-01-01

    Although Radio Frequency Identification (RFID) is poised to displace barcodes, security vulnerabilities pose serious challenges for global adoption of the RFID technology. Specifically, RFID tags are prone to basic cloning and counterfeiting security attacks. A successful cloning of the RFID tags in many commercial applications can lead to many serious problems such as financial losses, brand damage, safety and health of the public. With many industries such as pharmaceutical and businesses deploying RFID technology with a variety of products, it is important to tackle RFID tag cloning problem and improve the resistance of the RFID systems. To this end, we propose an approach for detecting cloned RFID tags in RFID systems with high detection accuracy and minimal overhead thus overcoming practical challenges in existing approaches. The proposed approach is based on consistency of dual hash collisions and modified count-min sketch vector. We evaluated the proposed approach through extensive experiments and compared it with existing baseline approaches in terms of execution time and detection accuracy under varying RFID tag cloning ratio. The results of the experiments show that the proposed approach outperforms the baseline approaches in cloned RFID tag detection accuracy.

  11. Use of the Nanofitin Alternative Scaffold as a GFP-Ready Fusion Tag

    PubMed Central

    Huet, Simon; Gorre, Harmony; Perrocheau, Anaëlle; Picot, Justine; Cinier, Mathieu

    2015-01-01

    With the continuous diversification of recombinant DNA technologies, the possibilities for new tailor-made protein engineering have extended on an on-going basis. Among these strategies, the use of the green fluorescent protein (GFP) as a fusion domain has been widely adopted for cellular imaging and protein localization. Following the lead of the direct head-to-tail fusion of GFP, we proposed to provide additional features to recombinant proteins by genetic fusion of artificially derived binders. Thus, we reported a GFP-ready fusion tag consisting of a small and robust fusion-friendly anti-GFP Nanofitin binding domain as a proof-of-concept. While limiting steric effects on the carrier, the GFP-ready tag allows the capture of GFP or its blue (BFP), cyan (CFP) and yellow (YFP) alternatives. Here, we described the generation of the GFP-ready tag from the selection of a Nanofitin variant binding to the GFP and its spectral variants with a nanomolar affinity, while displaying a remarkable folding stability, as demonstrated by its full resistance upon thermal sterilization process or the full chemical synthesis of Nanofitins. To illustrate the potential of the Nanofitin-based tag as a fusion partner, we compared the expression level in Escherichia coli and activity profile of recombinant human tumor necrosis factor alpha (TNFα) constructs, fused to a SUMO or GFP-ready tag. Very similar expression levels were found with the two fusion technologies. Both domains of the GFP-ready tagged TNFα were proved fully active in ELISA and interferometry binding assays, allowing the simultaneous capture by an anti-TNFα antibody and binding to the GFP, and its spectral mutants. The GFP-ready tag was also shown inert in a L929 cell based assay, demonstrating the potent TNFα mediated apoptosis induction by the GFP-ready tagged TNFα. Eventually, we proposed the GFP-ready tag as a versatile capture and labeling system in addition to expected applications of anti-GFP Nanofitins (as illustrated with previously described state-of-the-art anti-GFP binders applied to living cells and in vitro applications). Through a single fusion domain, the GFP-ready tagged proteins benefit from subsequent customization within a wide range of fluorescence spectra upon indirect binding of a chosen GFP variant. PMID:26539718

  12. Use of the Nanofitin Alternative Scaffold as a GFP-Ready Fusion Tag.

    PubMed

    Huet, Simon; Gorre, Harmony; Perrocheau, Anaëlle; Picot, Justine; Cinier, Mathieu

    2015-01-01

    With the continuous diversification of recombinant DNA technologies, the possibilities for new tailor-made protein engineering have extended on an on-going basis. Among these strategies, the use of the green fluorescent protein (GFP) as a fusion domain has been widely adopted for cellular imaging and protein localization. Following the lead of the direct head-to-tail fusion of GFP, we proposed to provide additional features to recombinant proteins by genetic fusion of artificially derived binders. Thus, we reported a GFP-ready fusion tag consisting of a small and robust fusion-friendly anti-GFP Nanofitin binding domain as a proof-of-concept. While limiting steric effects on the carrier, the GFP-ready tag allows the capture of GFP or its blue (BFP), cyan (CFP) and yellow (YFP) alternatives. Here, we described the generation of the GFP-ready tag from the selection of a Nanofitin variant binding to the GFP and its spectral variants with a nanomolar affinity, while displaying a remarkable folding stability, as demonstrated by its full resistance upon thermal sterilization process or the full chemical synthesis of Nanofitins. To illustrate the potential of the Nanofitin-based tag as a fusion partner, we compared the expression level in Escherichia coli and activity profile of recombinant human tumor necrosis factor alpha (TNFα) constructs, fused to a SUMO or GFP-ready tag. Very similar expression levels were found with the two fusion technologies. Both domains of the GFP-ready tagged TNFα were proved fully active in ELISA and interferometry binding assays, allowing the simultaneous capture by an anti-TNFα antibody and binding to the GFP, and its spectral mutants. The GFP-ready tag was also shown inert in a L929 cell based assay, demonstrating the potent TNFα mediated apoptosis induction by the GFP-ready tagged TNFα. Eventually, we proposed the GFP-ready tag as a versatile capture and labeling system in addition to expected applications of anti-GFP Nanofitins (as illustrated with previously described state-of-the-art anti-GFP binders applied to living cells and in vitro applications). Through a single fusion domain, the GFP-ready tagged proteins benefit from subsequent customization within a wide range of fluorescence spectra upon indirect binding of a chosen GFP variant.

  13. WebTag: Web browsing into sensor tags over NFC.

    PubMed

    Echevarria, Juan Jose; Ruiz-de-Garibay, Jonathan; Legarda, Jon; Alvarez, Maite; Ayerbe, Ana; Vazquez, Juan Ignacio

    2012-01-01

    Information and Communication Technologies (ICTs) continue to overcome many of the challenges related to wireless sensor monitoring, such as for example the design of smarter embedded processors, the improvement of the network architectures, the development of efficient communication protocols or the maximization of the life cycle autonomy. This work tries to improve the communication link of the data transmission in wireless sensor monitoring. The upstream communication link is usually based on standard IP technologies, but the downstream side is always masked with the proprietary protocols used for the wireless link (like ZigBee, Bluetooth, RFID, etc.). This work presents a novel solution (WebTag) for a direct IP based access to a sensor tag over the Near Field Communication (NFC) technology for secure applications. WebTag allows a direct web access to the sensor tag by means of a standard web browser, it reads the sensor data, configures the sampling rate and implements IP based security policies. It is, definitely, a new step towards the evolution of the Internet of Things paradigm.

  14. A UHF RFID system with on-chip-antenna tag for short range communication

    NASA Astrophysics Data System (ADS)

    Qi, Peng; Chun, Zhang; Xijin, Zhao; Zhihua, Wang

    2015-05-01

    A UHF RF identification system based on the 0.18 μm CMOS process has been developed for short range and harsh size requirement applications, which is composed of a fully integrated tag and a special reader. The whole tag chip with the antenna takes up an area of 0.36 mm2, which is smaller than other reported tags with an on-chip antenna (OCA) using the standard CMOS process. A self-defined protocol is proposed to reduce the power consumption, and minimize the size of the tag. The specialized SOC reader system consists of the RF transceiver, digital baseband, MCU and host interface. Its power consumption is about 500 mW. Measurement results show that the system's reading range is 2 mm with 20 dBm reader output power. With an inductive antenna printed on a paper substrate around the OCA tag, the reading range can be extended from several centimeters to meters, depending on the shape and size of the inductive antenna.

  15. Enantioselective chromatography in analysis of triacylglycerols common in edible fats and oils.

    PubMed

    Kalpio, Marika; Nylund, Matts; Linderborg, Kaisa M; Yang, Baoru; Kristinsson, Björn; Haraldsson, Gudmundur G; Kallio, Heikki

    2015-04-01

    Enantiomers of racemic triacylglycerol (TAG) mixtures were separated using two chiral HPLC columns with a sample recycling system and a UV detector. A closed system without sample derivatisation enabled separation and identification by using enantiopure reference compounds of eleven racemic TAGs with C12-C22 fatty acids with 0-2 double bonds. The prolonged separation time was compensated for by fewer pretreatment steps. Presence of one saturated and one unsaturated fatty acid in the asymmetric TAG favoured the separation. Enantiomeric resolution, at the same time with stronger retention of TAGs, increased with increasing fatty acid chain length in the sn-1(3) position. Triunsaturated TAGs containing oleic, linoleic or palmitoleic acids did not separate. The elution order of enantiomers was determined by chemoenzymatically synthesised enantiopure TAGs with a co-injection method. The method is applicable to many natural fats and oils of low unsaturation level assisting advanced investigation of lipid synthesis and metabolism. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. WebTag: Web Browsing into Sensor Tags over NFC

    PubMed Central

    Echevarria, Juan Jose; Ruiz-de-Garibay, Jonathan; Legarda, Jon; Álvarez, Maite; Ayerbe, Ana; Vazquez, Juan Ignacio

    2012-01-01

    Information and Communication Technologies (ICTs) continue to overcome many of the challenges related to wireless sensor monitoring, such as for example the design of smarter embedded processors, the improvement of the network architectures, the development of efficient communication protocols or the maximization of the life cycle autonomy. This work tries to improve the communication link of the data transmission in wireless sensor monitoring. The upstream communication link is usually based on standard IP technologies, but the downstream side is always masked with the proprietary protocols used for the wireless link (like ZigBee, Bluetooth, RFID, etc.). This work presents a novel solution (WebTag) for a direct IP based access to a sensor tag over the Near Field Communication (NFC) technology for secure applications. WebTag allows a direct web access to the sensor tag by means of a standard web browser, it reads the sensor data, configures the sampling rate and implements IP based security policies. It is, definitely, a new step towards the evolution of the Internet of Things paradigm. PMID:23012511

  17. Mild pressure induces rapid accumulation of neutral lipid (triacylglycerol) in Chlorella spp.

    PubMed

    Praveenkumar, Ramasamy; Kim, Bohwa; Lee, Jiye; Vijayan, Durairaj; Lee, Kyubock; Nam, Bora; Jeon, Sang Goo; Kim, Dong-Myung; Oh, You-Kwan

    2016-11-01

    Effective enhancement of neutral lipid (especially triacylglycerol, TAG) content in microalgae is an important issue for commercialization of microalgal biorefineries. Pressure is a key physical factor affecting the morphological, physiological, and biochemical behaviors of organisms. In this paper, we report a new stress-based method for induction of TAG accumulation in microalgae (specifically, Chlorella sp. KR-1 and Ch. sp. AG20150) by very-short-duration application of mild pressure. Pressure treatments of 10-15bar for 2h resulted in a considerable, ∼55% improvement of the 10-100g/Lcells' TAG contents compared with the untreated control. The post-pressure-treatment increase of cytoplasmic TAG granules was further confirmed by transmission electron microscopy (TEM). Notwithstanding the increased TAG content, the total lipid content was not changed by pressurization, implying that pressure stress possibly induces rapid remodeling/transformation of algal lipids rather than de novo biosynthesis of TAG. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Expression and affinity purification of recombinant proteins from plants

    NASA Technical Reports Server (NTRS)

    Desai, Urvee A.; Sur, Gargi; Daunert, Sylvia; Babbitt, Ruth; Li, Qingshun

    2002-01-01

    With recent advances in plant biotechnology, transgenic plants have been targeted as an inexpensive means for the mass production of proteins for biopharmaceutical and industrial uses. However, the current plant purification techniques lack a generally applicable, economic, large-scale strategy. In this study, we demonstrate the purification of a model protein, beta-glucuronidase (GUS), by employing the protein calmodulin (CaM) as an affinity tag. In the proposed system, CaM is fused to GUS. In the presence of calcium, the calmodulin fusion protein binds specifically to a phenothiazine-modified surface of an affinity column. When calcium is removed with a complexing agent, e.g., EDTA, calmodulin undergoes a conformational change allowing the dissociation of the calmodulin-phenothiazine complex and, therefore, permitting the elution of the GUS-CaM fusion protein. The advantages of this approach are the fast, efficient, and economical isolation of the target protein under mild elution conditions, thus preserving the activity of the target protein. Two types of transformation methods were used in this study, namely, the Agrobacterium-mediated system and the viral-vector-mediated transformation system. Copyright 2002 Elsevier Science (USA).

  19. Alteration of gene expression by restriction enzymes electroporated into plant cells.

    PubMed

    Ashraf, M; Altschuler, M; Galasinski, S; Griffiths, T D

    1993-06-01

    The alteration in the expression of a beta-glucuronidase (GUS) reporter gene was used to monitor the effect of restriction endonucleases electroporated into the tobacco (Nicotiana tabacum L.) protoplasts. Restriction enzyme (RE) Hind III which does not have a recognition site within the gene cassette, had little effect on enzyme activity. In contrast restriction endonucleases Hae III and Sau3A1 which possess 8 and 16 recognition sites in the GUS cassette, were found to reduce the enzyme activity by 89% and 94% respectively when compared to control electroporations. Restriction-site mutation analysis (RSM) and Southern blot analysis indicated the enzymatic degradation of GUS coding sequence by the REs Hae III and Sau3A1. Results of this study suggest that on electroporation, REs can enter into plant cells and alter the expression of the GUS gene. The alteration of gene expression is thus correlated with the digestion of GUS template DNA. Future applications of this technique could include addressing fundamental questions with regard to DNA repair, site-specific recombination, identifying mutations, insertional mutagenesis, enhancement of stable transformation and gene tagging in plants.

  20. VEGF elicits epithelial-mesenchymal transition (EMT) in prostate intraepithelial neoplasia (PIN)-like cells via an autocrine loop

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gonzalez-Moreno, Oscar; Lecanda, Jon; Green, Jeffrey E.

    2010-02-15

    Vascular endothelial growth factor (VEGF) is overexpressed during the transition from prostate intraepithelial neoplasia (PIN) to invasive carcinoma. We have mimicked such a process in vitro using the PIN-like C3(1)/Tag-derived Pr-111 cell line, which expresses low levels of VEGF and exhibits very low tumorigenicity in vivo. Elevated expression of VEGF164 in Pr-111 cells led to a significant increase in tumorigenicity, invasiveness, proliferation rates and angiogenesis. Moreover, VEGF164 induced strong changes in cell morphology and cell transcriptome through an autocrine mechanism, with changes in TGF-beta1- and cytoskeleton-related pathways, among others. Further analysis of VEGF-overexpressing Pr-111 cells or following exogenous addition ofmore » recombinant VEGF shows acquisition of epithelial-mesenchymal transition (EMT) features, with an increased expression of mesenchymal markers, such as N-cadherin, Snail1, Snail2 (Slug) and vimentin, and a decrease in E-cadherin. Administration of VEGF led to changes in TGF-beta1 signaling, including reduction of Smad7 (TGF-beta inhibitory Smad), increase in TGF-betaR-II, and translocation of phospho-Smad3 to the nucleus. Our results suggest that increased expression of VEGF in malignant cells during the transition from PIN to invasive carcinoma leads to EMT through an autocrine loop, which would promote tumor cell invasion and motility. Therapeutic blockade of VEGF/TGF-beta1 in PIN lesions might impair not only tumor angiogenesis, but also the early dissemination of malignant cells outside the epithelial layer.« less

  1. Metabolic engineering of Saccharomyces cerevisiae to produce a reduced viscosity oil from lignocellulose

    DOE PAGES

    Tran, Tam N. T.; Breuer, Rebecca J.; Avanasi Narasimhan, Ragothaman; ...

    2017-03-20

    Background: Acetyl-triacylglycerols (acetyl-TAGs) are unusual triacylglycerol (TAG) molecules that contain an sn-3 acetate group. Compared to typical triacylglycerol molecules (here referred to as long chain TAGs; lcTAGs), acetyl-TAGs possess reduced viscosity and improved cold temperature properties, which may allow direct use as a drop-in diesel fuel. Their different chemical and physical properties also make acetyl-TAGs useful for other applications such as lubricants and plasticizers. Acetyl-TAGs can be synthesized by EaDAcT, a diacylglycerol acetyltransferase enzyme originally isolated from Euonymus alatus (Burning Bush). The heterologous expression of EaDAcT in different organisms, including Saccharomyces cerevisiae, resulted in the accumulation of acetyl-TAGs in storagemore » lipids. Microbial conversion of lignocellulose into acetyl-TAGs could allow biorefinery production of versatile molecules for biofuel and bioproducts. Results: In order to produce acetyl-TAGs from abundant lignocellulose feedstocks, we expressed EaDAcT in S. cerevisiae previously engineered to utilize xylose as a carbon source. The resulting strains were capable of producing acetyl-TAGs when grown on different media. The highest levels of acetyl-TAG production were observed with growth on synthetic lab media containing glucose or xylose. Importantly, acetyl-TAGs were also synthesized by this strain in ammonia fiber expansion (AFEX)-pretreated corn stover hydrolysate (ACSH) at higher volumetric titers than previously published strains. The deletion of the four endogenous enzymes known to contribute to lcTAG production increased the proportion of acetyl-TAGs in the total storage lipids beyond that in existing strains, which will make purification of these useful lipids easier. Surprisingly, the strains containing the four deletions were still capable of synthesizing lcTAG, suggesting that the particular strain used in this study possesses additional undetermined diacylglycerol acyltransferase activity. Additionally, the carbon source used for growth influenced the accumulation of these residual lcTAGs, with higher levels in strains cultured on xylose containing media. Conclusion: Our results demonstrate that S. cerevisiae can be metabolically engineered to produce acetyl-TAGs when grown on different carbon sources, including hydrolysate derived from lignocellulose. Deletion of four endogenous acyltransferases enabled a higher purity of acetyl-TAGs to be achieved, but lcTAGs were still synthesized. Longer incubation times also decreased the levels of acetyl-TAGs produced. Therefore, additional work is needed to further manipulate acetyl-TAG production in this strain of S. cerevisiae, including the identification of other TAG biosynthetic and lipolytic enzymes and a better understanding of the regulation of the synthesis and degradation of storage lipids.« less

  2. A study on PubMed search tag usage pattern: association rule mining of a full-day PubMed query log.

    PubMed

    Mosa, Abu Saleh Mohammad; Yoo, Illhoi

    2013-01-09

    The practice of evidence-based medicine requires efficient biomedical literature search such as PubMed/MEDLINE. Retrieval performance relies highly on the efficient use of search field tags. The purpose of this study was to analyze PubMed log data in order to understand the usage pattern of search tags by the end user in PubMed/MEDLINE search. A PubMed query log file was obtained from the National Library of Medicine containing anonymous user identification, timestamp, and query text. Inconsistent records were removed from the dataset and the search tags were extracted from the query texts. A total of 2,917,159 queries were selected for this study issued by a total of 613,061 users. The analysis of frequent co-occurrences and usage patterns of the search tags was conducted using an association mining algorithm. The percentage of search tag usage was low (11.38% of the total queries) and only 2.95% of queries contained two or more tags. Three out of four users used no search tag and about two-third of them issued less than four queries. Among the queries containing at least one tagged search term, the average number of search tags was almost half of the number of total search terms. Navigational search tags are more frequently used than informational search tags. While no strong association was observed between informational and navigational tags, six (out of 19) informational tags and six (out of 29) navigational tags showed strong associations in PubMed searches. The low percentage of search tag usage implies that PubMed/MEDLINE users do not utilize the features of PubMed/MEDLINE widely or they are not aware of such features or solely depend on the high recall focused query translation by the PubMed's Automatic Term Mapping. The users need further education and interactive search application for effective use of the search tags in order to fulfill their biomedical information needs from PubMed/MEDLINE.

  3. A Study on Pubmed Search Tag Usage Pattern: Association Rule Mining of a Full-day Pubmed Query Log

    PubMed Central

    2013-01-01

    Background The practice of evidence-based medicine requires efficient biomedical literature search such as PubMed/MEDLINE. Retrieval performance relies highly on the efficient use of search field tags. The purpose of this study was to analyze PubMed log data in order to understand the usage pattern of search tags by the end user in PubMed/MEDLINE search. Methods A PubMed query log file was obtained from the National Library of Medicine containing anonymous user identification, timestamp, and query text. Inconsistent records were removed from the dataset and the search tags were extracted from the query texts. A total of 2,917,159 queries were selected for this study issued by a total of 613,061 users. The analysis of frequent co-occurrences and usage patterns of the search tags was conducted using an association mining algorithm. Results The percentage of search tag usage was low (11.38% of the total queries) and only 2.95% of queries contained two or more tags. Three out of four users used no search tag and about two-third of them issued less than four queries. Among the queries containing at least one tagged search term, the average number of search tags was almost half of the number of total search terms. Navigational search tags are more frequently used than informational search tags. While no strong association was observed between informational and navigational tags, six (out of 19) informational tags and six (out of 29) navigational tags showed strong associations in PubMed searches. Conclusions The low percentage of search tag usage implies that PubMed/MEDLINE users do not utilize the features of PubMed/MEDLINE widely or they are not aware of such features or solely depend on the high recall focused query translation by the PubMed’s Automatic Term Mapping. The users need further education and interactive search application for effective use of the search tags in order to fulfill their biomedical information needs from PubMed/MEDLINE. PMID:23302604

  4. Energy-Aware RFID Anti-Collision Protocol.

    PubMed

    Arjona, Laura; Simon, Hugo Landaluce; Ruiz, Asier Perallos

    2018-06-11

    The growing interest in mobile devices is transforming wireless identification technologies. Mobile and battery-powered Radio Frequency Identification (RFID) readers, such as hand readers and smart phones, are are becoming increasingly attractive. These RFID readers require energy-efficient anti-collision protocols to minimize the tag collisions and to expand the reader's battery life. Furthermore, there is an increasing interest in RFID sensor networks with a growing number of RFID sensor tags. Thus, RFID application developers must be mindful of tag anti-collision protocols. Energy-efficient protocols involve a low reader energy consumption per tag. This work presents a thorough study of the reader energy consumption per tag and analyzes the main factor that affects this metric: the frame size update strategy. Using the conclusion of this analysis, the anti-collision protocol Energy-Aware Slotted Aloha (EASA) is presented to decrease the energy consumption per tag. The frame size update strategy of EASA is configured to minimize the energy consumption per tag. As a result, EASA presents an energy-aware frame. The performance of the proposed protocol is evaluated and compared with several state of the art Aloha-based anti-collision protocols based on the current RFID standard. Simulation results show that EASA, with an average of 15 mJ consumed per tag identified, achieves a 6% average improvement in the energy consumption per tag in relation to the strategies of the comparison.

  5. Array processing for RFID tag localization exploiting multi-frequency signals

    NASA Astrophysics Data System (ADS)

    Zhang, Yimin; Li, Xin; Amin, Moeness G.

    2009-05-01

    RFID is an increasingly valuable business and technology tool for electronically identifying, locating, and tracking products, assets, and personnel. As a result, precise positioning and tracking of RFID tags and readers have received considerable attention from both academic and industrial communities. Finding the position of RFID tags is considered an important task in various real-time locating systems (RTLS). As such, numerous RFID localization products have been developed for various applications. The majority of RFID positioning systems is based on the fusion of pieces of relevant information, such as the range and the direction-of-arrival (DOA). For example, trilateration can determine the tag position by using the range information of the tag estimated from three or more spatially separated reader antennas. Triangulation is another method to locate RFID tags that use the direction-of-arrival (DOA) information estimated at multiple spatially separated locations. The RFID tag positions can also be determined through hybrid techniques that combine the range and DOA information. The focus of this paper to study the design and performance of the localization of passive RFID tags using array processing techniques in a multipath environment, and exploiting multi-frequency CW signals. The latter are used to decorrelate the coherent multipath signals for effective DOA estimation and for the purpose of accurate range estimation. Accordingly, the spatial and frequency dimensionalities are fully utilized for robust and accurate positioning of RFID tags.

  6. Serotype determination of Salmonella by xTAG assay.

    PubMed

    Zheng, Zhibei; Zheng, Wei; Wang, Haoqiu; Pan, Jincao; Pu, Xiaoying

    2017-10-01

    Currently, no protocols or commercial kits are available to determine the serotypes of Salmonella by using Luminex MAGPIX®. In this study, an xTAG assay for serotype determination of Salmonella suitable for Luminex MAGPIX® is described and 228 Salmonella isolates were serotype determined by this xTAG assay. The xTAG assay consists of two steps: 1) Multiplex PCR to amplify simultaneously O, H and Vi antigen genes of Salmonella, and 2) Magplex-TAG™ microsphere hybridization to identify accurately the specific PCR products of different antigens. Compared with the serotyping results of traditional serum agglutination test, the sensitivity and specificity of the xTAG assay were 95.1% and 100%, respectively. The agreement rate of these two assays was 95.2%. Compared with Luminex xMAP® Salmonella Serotyping Assay (SSA) kit, the advantages of this xTAG assay are: First, the magnetic beads make it applicable to both the Luminex®100/200™ and MAGPIX® systems. Second, only primers rather than both primers and probes are needed in the xTAG assay, and the process of coupling antigen-specific oligonucleotide probes to beads is circumvented, which make the xTAG assay convenient to be utilized by other laboratories. The xTAG assay may serve as a rapid alternative or complementary method for traditional Salmonella serotyping tests, especially for laboratories that utilize the MAGPIX® systems. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Generation and esterification of electrophilic fatty acid nitroalkenes in triacylglycerides

    PubMed Central

    Fazzari, Marco; Khoo, Nicholas; Woodcock, Steven R.; Li, Lihua; Freeman, Bruce A.; Schopfer, Francisco J.

    2015-01-01

    Electrophilic fatty acid nitroalkenes (NO2-FA) are products of nitric oxide and nitrite-mediated unsaturated fatty acid nitration. These electrophilic products induce pleiotropic signaling actions that modulate metabolic and inflammatory responses in cell and animal models. The metabolism of NO2-FA includes reduction of the vinyl nitro moiety by prostaglandin reductase-1, mitochondrial β–oxidation and Michael addition with low molecular weight nucleophilic amino acids. Complex lipid reactions of fatty acid nitroalkenes are not well defined. Herein we report the detection and characterization of NO2-FA-containing triacylglycerides (NO2-FA-TAG) via mass spectrometry-based methods. In this regard, unsaturated fatty acids of dietary triacylglycerides are targets for nitration reactions during gastric acidification, where NO2-FA-TAG can be detected in rat plasma after oral administration of nitro-oleic acid (NO2-OA). Furthermore, the characterization and profiling of these species, including the generation of beta oxidation and dehydrogenation products, could be detected in NO2-OA supplemented adipocytes. These data revealed that NO2-FA-TAG, formed by either the direct nitration of esterified unsaturated fatty acids or the incorporation of nitrated free fatty acids into triacylglycerides, contribute to the systemic distribution of these reactive electrophilic mediators and may serve as a depot for subsequent mobilization by lipases to in turn impact adipocyte homeostasis and tissue signaling events. PMID:26066303

  8. Focal complex formation in adult cardiomyocytes is accompanied by the activation of beta3 integrin and c-Src.

    PubMed

    Willey, Christopher D; Balasubramanian, Sundaravadivel; Rodríguez Rosas, María C; Ross, Robert S; Kuppuswamy, Dhandapani

    2003-06-01

    In pressure-overloaded myocardium, our recent study demonstrated cytoskeletal assembly of c-Src and other signaling proteins which was partially mimicked in vitro using adult feline cardiomyocytes embedded in three-dimensional (3D) collagen matrix and stimulated with an integrin-binding Arg-Gly-Asp (RGD) peptide. In the present study, we improved this model further to activate c-Src and obtain a full assembly of the focal adhesion complex (FAC), and characterized c-Src localization and integrin subtype(s) involved. RGD dose response experiments revealed that c-Src activation occurs subsequent to its cytoskeletal recruitment and is accompanied by p130Cas cytoskeletal binding and focal adhesion kinase (FAK) Tyr925 phosphorylation. When cardiomyocytes expressing hexahistidine-tagged c-Src via adenoviral gene delivery were used for RGD stimulation, the expressed c-Src exhibited relocation: (i) biochemical analysis revealed c-Src movement from the detergent-soluble to the -insoluble cytoskeletal fraction and (ii) confocal microscopic analysis showed c-Src movement from a nuclear/perinuclear to a sarcolemmal region. RGD treatment also caused sarcolemmal co-localization of FAK and vinculin. Characterization of integrin subtypes revealed that beta3, but not beta1, integrin plays a predominant role: (i) expression of cytoplasmic domain of beta1A integrin did not affect the RGD-stimulated FAC formation and (ii) both pressure-overloaded myocardium and RGD-stimulated cardiomyocytes exhibited phosphorylation of beta3 integrin at Tyr773/785 sites but not beta1 integrin at Thr788/789 sites. Together these data indicate that RGD treatment in cardiomyocytes causes beta3 integrin activation and c-Src sarcolemmal localization, that subsequent c-Src activation is accompanied by p130Cas binding and FAK Tyr925 phosphorylation, and that these events might be crucial for growth and remodeling of hypertrophying adult cardiomyocytes.

  9. Superior triacylglycerol (TAG) accumulation in starchless mutants of Scenedesmus obliquus: (I) mutant generation and characterization

    PubMed Central

    2014-01-01

    Background Microalgae are a promising platform for producing neutral lipids, to be used in the application for biofuels or commodities in the feed and food industry. A very promising candidate is the oleaginous green microalga Scenedesmus obliquus, because it accumulates up to 45% w/w triacylglycerol (TAG) under nitrogen starvation. Under these conditions, starch is accumulated as well. Starch can amount up to 38% w/w under nitrogen starvation, which is a substantial part of the total carbon captured. When aiming for optimized TAG production, blocking the formation of starch could potentially increase carbon allocation towards TAG. In an attempt to increase TAG content, productivity and yield, starchless mutants of this high potential strain were generated using UV mutagenesis. Previous studies in Chlamydomonas reinhardtii have shown that blocking the starch synthesis yields higher TAG contents, although these TAG contents do not surpass those of oleaginous microalgae yet. So far no starchless mutants in oleaginous green microalgae have been isolated that result in higher TAG productivities. Results Five starchless mutants have been isolated successfully from over 3,500 mutants. The effect of the mutation on biomass and total fatty acid (TFA) and TAG productivity under nitrogen-replete and nitrogen-depleted conditions was studied. All five starchless mutants showed a decreased or completely absent starch content. In parallel, an increased TAG accumulation rate was observed for the starchless mutants and no substantial decrease in biomass productivity was perceived. The most promising mutant showed an increase in TFA productivity of 41% at 4 days after nitrogen depletion, reached a TAG content of 49.4% (% of dry weight) and had no substantial change in biomass productivity compared to the wild type. Conclusions The improved S. obliquus TAG production strains are the first starchless mutants in an oleaginous green microalga that show enhanced TAG content under photoautotrophic conditions. These results can pave the way towards a more feasible microalgae-driven TAG production platform. PMID:24920957

  10. Triacylglycerol is produced from starch and polar lipids in the green alga Dunaliella tertiolecta.

    PubMed

    Pick, Uri; Avidan, Omri

    2017-10-13

    The halotolerant green alga Dunaliella tertiolecta accumulates starch and triacylglycerol (TAG) amounting to 70% and 10-15% of total cellular carbon, respectively, when exposed to nitrogen (N) deprivation. The purpose of this study was to clarify the inter-relationships between the biosynthesis of TAG, starch, and polar lipids (PLs) in this alga. Pulse labeling with [14C]bicarbonate was utilized to label starch and [14C]palmitic acid (PlA) to label lipids. Transfer of 14C into TAG was measured and used to calculate rates of synthesis. About two-thirds of the carbon in TAG originates from starch, and one-third is made de novo by direct CO2 assimilation. The level made from degradation of pre-formed PLs is estimated to be very small. Most of the de novo synthesis involves fatty acid transfer through PLs made during the first day of N deprivation. The results suggest that starch made by photosynthetic carbon assimilation at the early stages of N deprivation is utilized for synthesis of TAG. Trans-acylation from PLs is the second major contributor to TAG biosynthesis. The utilization of starch for TAG biosynthesis may have biotechnological applications to optimize TAG biosynthesis in algae. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  11. Dynamics of human categorization in a collaborative tagging system: How social processes of semantic stabilization shape individual sensemaking.

    PubMed

    Ley, Tobias; Seitlinger, Paul

    2015-10-01

    We study how categories form and develop over time in a sensemaking task by groups of students employing a collaborative tagging system. In line with distributed cognition theories, we look at both the tags students use and their strength of representation in memory. We hypothesize that categories get more differentiated over time as students learn, and that semantic stabilization on the group level (i.e. the convergence in the use of tags) mediates this relationship. Results of a field experiment that tested the impact of topic study duration on the specificity of tags confirms these hypotheses, although it was not study duration that produced this effect, but rather the effectiveness of the collaborative taxonomy the groups built. In the groups with higher levels of semantic stabilization, we found use of more specific tags and better representation in memory. We discuss these findings with regard to the important role of the information value of tags that would drive both the convergence on the group level as well as a shift to more specific levels of categorization. We also discuss the implication for cognitive science research by highlighting the importance of collaboratively built artefacts in the process of how knowledge is acquired, and implications for educational applications of collaborative tagging environments.

  12. Dynamics of human categorization in a collaborative tagging system: How social processes of semantic stabilization shape individual sensemaking

    PubMed Central

    Ley, Tobias; Seitlinger, Paul

    2015-01-01

    We study how categories form and develop over time in a sensemaking task by groups of students employing a collaborative tagging system. In line with distributed cognition theories, we look at both the tags students use and their strength of representation in memory. We hypothesize that categories get more differentiated over time as students learn, and that semantic stabilization on the group level (i.e. the convergence in the use of tags) mediates this relationship. Results of a field experiment that tested the impact of topic study duration on the specificity of tags confirms these hypotheses, although it was not study duration that produced this effect, but rather the effectiveness of the collaborative taxonomy the groups built. In the groups with higher levels of semantic stabilization, we found use of more specific tags and better representation in memory. We discuss these findings with regard to the important role of the information value of tags that would drive both the convergence on the group level as well as a shift to more specific levels of categorization. We also discuss the implication for cognitive science research by highlighting the importance of collaboratively built artefacts in the process of how knowledge is acquired, and implications for educational applications of collaborative tagging environments. PMID:26566299

  13. In situ tagging technique for fishes provides insight into growth and movement of invasive lionfish

    PubMed Central

    Akins, John L; Morris, James A; Green, Stephanie J

    2014-01-01

    Information on fish movement and growth is primarily obtained through the marking and tracking of individuals with external tags, which are usually affixed to anesthetized individuals at the surface. However, the quantity and quality of data obtained by this method is often limited by small sample sizes owing to the time associated with the tagging process, high rates of tagging-related mortality, and displacement of tagged individuals from the initial capture location. To address these issues, we describe a technique for applying external streamer and dart tags in situ, which uses SCUBA divers to capture and tag individual fish on the sea floor without the use of anesthetic. We demonstrate this method for Indo-Pacific lionfish (Pterois volitans/P. miles), species which are particularly vulnerable to barotrauma when transported to and handled at the surface. To test our method, we tagged 161 individuals inhabiting 26 coral reef locations in the Bahamas over a period of 3 years. Our method resulted in no instances of barotrauma, reduced handling and recovery time, and minimal post-tagging release displacement compared with conventional ex situ tag application. Opportunistic resighting and recapture of tagged individuals reveals that lionfish exhibit highly variable site fidelity, movement patterns, and growth rates on invaded coral reef habitats. In total, 24% of lionfish were resighted between 29 and 188 days after tagging. Of these, 90% were located at the site of capture, while the remaining individuals were resighted between 200 m and 1.1 km from initial site of capture over 29 days later. In situ growth rates ranged between 0.1 and 0.6 mm/day. While individuals tagged with streamer tags posted slower growth rates with increasing size, as expected, there was no relationship between growth rate and fish size for individuals marked with dart tags, potentially because of large effects of tag presence on the activities of small bodied lionfish (i.e., <150 mm), where the tag was up to 7.6% of the lionfish's mass. Our study offers a novel in situ tagging technique that can be used to provide critical information on fish site fidelity, movement patterns, and growth in cases where ex situ tagging is not feasible. PMID:25614791

  14. In situ tagging technique for fishes provides insight into growth and movement of invasive lionfish.

    PubMed

    Akins, John L; Morris, James A; Green, Stephanie J

    2014-10-01

    Information on fish movement and growth is primarily obtained through the marking and tracking of individuals with external tags, which are usually affixed to anesthetized individuals at the surface. However, the quantity and quality of data obtained by this method is often limited by small sample sizes owing to the time associated with the tagging process, high rates of tagging-related mortality, and displacement of tagged individuals from the initial capture location. To address these issues, we describe a technique for applying external streamer and dart tags in situ, which uses SCUBA divers to capture and tag individual fish on the sea floor without the use of anesthetic. We demonstrate this method for Indo-Pacific lionfish (Pterois volitans/P. miles), species which are particularly vulnerable to barotrauma when transported to and handled at the surface. To test our method, we tagged 161 individuals inhabiting 26 coral reef locations in the Bahamas over a period of 3 years. Our method resulted in no instances of barotrauma, reduced handling and recovery time, and minimal post-tagging release displacement compared with conventional ex situ tag application. Opportunistic resighting and recapture of tagged individuals reveals that lionfish exhibit highly variable site fidelity, movement patterns, and growth rates on invaded coral reef habitats. In total, 24% of lionfish were resighted between 29 and 188 days after tagging. Of these, 90% were located at the site of capture, while the remaining individuals were resighted between 200 m and 1.1 km from initial site of capture over 29 days later. In situ growth rates ranged between 0.1 and 0.6 mm/day. While individuals tagged with streamer tags posted slower growth rates with increasing size, as expected, there was no relationship between growth rate and fish size for individuals marked with dart tags, potentially because of large effects of tag presence on the activities of small bodied lionfish (i.e., <150 mm), where the tag was up to 7.6% of the lionfish's mass. Our study offers a novel in situ tagging technique that can be used to provide critical information on fish site fidelity, movement patterns, and growth in cases where ex situ tagging is not feasible.

  15. Life and Death of Deep-Sea Vents: Bacterial Diversity and Ecosystem Succession on Inactive Hydrothermal Sulfides

    PubMed Central

    Sylvan, Jason B.; Toner, Brandy M.; Edwards, Katrina J.

    2012-01-01

    ABSTRACT Hydrothermal chimneys are a globally dispersed habitat on the seafloor associated with mid-ocean ridge (MOR) spreading centers. Active, hot, venting sulfide structures from MORs have been examined for microbial diversity and ecology since their discovery in the mid-1970s, and recent work has also begun to explore the microbiology of inactive sulfides—structures that persist for decades to millennia and form moderate to massive deposits at and below the seafloor. Here we used tag pyrosequencing of the V6 region of the 16S rRNA and full-length 16S rRNA sequencing on inactive hydrothermal sulfide chimney samples from 9°N on the East Pacific Rise to learn their bacterial composition, metabolic potential, and succession from venting to nonventing (inactive) regimes. Alpha-, beta-, delta-, and gammaproteobacteria and members of the phylum Bacteroidetes dominate all inactive sulfides. Greater than 26% of the V6 tags obtained are closely related to lineages involved in sulfur, nitrogen, iron, and methane cycling. Epsilonproteobacteria represent <4% of the V6 tags recovered from inactive sulfides and 15% of the full-length clones, despite their high abundance in active chimneys. Members of the phylum Aquificae, which are common in active vents, were absent from both the V6 tags and full-length 16S rRNA data sets. In both analyses, the proportions of alphaproteobacteria, betaproteobacteria, and members of the phylum Bacteroidetes were greater than those found on active hydrothermal sulfides. These shifts in bacterial population structure on inactive chimneys reveal ecological succession following cessation of venting and also imply a potential shift in microbial activity and metabolic guilds on hydrothermal sulfides, the dominant biome that results from seafloor venting. PMID:22275502

  16. High Throughput Biological Analysis Using Multi-bit Magnetic Digital Planar Tags

    NASA Astrophysics Data System (ADS)

    Hong, B.; Jeong, J.-R.; Llandro, J.; Hayward, T. J.; Ionescu, A.; Trypiniotis, T.; Mitrelias, T.; Kopper, K. P.; Steinmuller, S. J.; Bland, J. A. C.

    2008-06-01

    We report a new magnetic labelling technology for high-throughput biomolecular identification and DNA sequencing. Planar multi-bit magnetic tags have been designed and fabricated, which comprise a magnetic barcode formed by an ensemble of micron-sized thin film Ni80Fe20 bars encapsulated in SU8. We show that by using a globally applied magnetic field and magneto-optical Kerr microscopy the magnetic elements in the multi-bit magnetic tags can be addressed individually and encoded/decoded remotely. The critical steps needed to show the feasibility of this technology are demonstrated, including fabrication, flow transport, remote writing and reading, and successful functionalization of the tags as verified by fluorescence detection. This approach is ideal for encoding information on tags in microfluidic flow or suspension, for such applications as labelling of chemical precursors during drug synthesis and combinatorial library-based high-throughput multiplexed bioassays.

  17. Conversion of red fluorescent protein into a bright blue probe.

    PubMed

    Subach, Oksana M; Gundorov, Illia S; Yoshimura, Masami; Subach, Fedor V; Zhang, Jinghang; Grüenwald, David; Souslova, Ekaterina A; Chudakov, Dmitriy M; Verkhusha, Vladislav V

    2008-10-20

    We used a red chromophore formation pathway, in which the anionic red chromophore is formed from the neutral blue intermediate, to suggest a rational design strategy to develop blue fluorescent proteins with a tyrosine-based chromophore. The strategy was applied to red fluorescent proteins of the different genetic backgrounds, such as TagRFP, mCherry, HcRed1, M355NA, and mKeima, which all were converted into blue probes. Further improvement of the blue variant of TagRFP by random mutagenesis resulted in an enhanced monomeric protein, mTagBFP, characterized by the substantially higher brightness, the faster chromophore maturation, and the higher pH stability than blue fluorescent proteins with a histidine in the chromophore. The detailed biochemical and photochemical analysis indicates that mTagBFP is the true monomeric protein tag for multicolor and lifetime imaging, as well as the outstanding donor for green fluorescent proteins in Förster resonance energy transfer applications.

  18. Quantification of Cell-Penetrating Peptide Associated with Polymeric Nanoparticles Using Isobaric-Tagging and MALDI-TOF MS/MS

    NASA Astrophysics Data System (ADS)

    Chiu, Jasper Z. S.; Tucker, Ian G.; McDowell, Arlene

    2016-11-01

    High sensitivity quantification of the putative cell-penetrating peptide di-arginine-histidine (RRH) associated with poly (ethyl-cyanoacrylate) (PECA) nanoparticles was achieved without analyte separation, using a novel application of isobaric-tagging and high matrix-assisted laser desorption/ionization coupled to time-of-flight (MALDI-TOF) mass spectrometry. Isobaric-tagging reaction equilibrium was reached after 5 min, with 90% or greater RRH peptide successfully isobaric-tagged after 60 min. The accuracy was greater than 90%, which indicates good reliability of using isobaric-tagged RRH as an internal standard for RRH quantification. The sample intra- and inter-spot coefficients of variations were less than 11%, which indicate good repeatability. The majority of RRH peptides in the nanoparticle formulation were physically associated with the nanoparticles (46.6%), whereas only a small fraction remained unassociated (13.7%). The unrecovered RRH peptide (~40%) was assumed to be covalently associated with PECA nanoparticles.

  19. The use of tags and tag clouds to discern credible content in online health message forums.

    PubMed

    O'Grady, Laura; Wathen, C Nadine; Charnaw-Burger, Jill; Betel, Lisa; Shachak, Aviv; Luke, Robert; Hockema, Stephen; Jadad, Alejandro R

    2012-01-01

    Web sites with health-oriented content are potentially harmful if inaccurate or inappropriate medical information is used to make health-related decisions. Checklists, rating systems and guidelines have been developed to help people determine what is credible, but recent Internet technologies emphasize applications that are collaborative in nature, including tags and tag clouds, where site users 'tag' or label online content, each using their own labelling system. Concepts such as the date, reference, author, testimonial and quotations are considered predictors of credible content. An understanding of these descriptive tools, how they relate to the depiction of credibility and how this relates to overall efforts to label data in relation to the semantic web has yet to emerge. This study investigates how structured (pre-determined) and unstructured (user-generated) tags and tag clouds with a multiple word search feature are used by participants to assess credibility of messages posted in online message forums. The targeted respondents were those using web sites message forums for disease self-management. We also explored the relevancy of our findings to the labelling or indexing of data in the context of the semantic web. Diabetes was chosen as the content area in this study, since (a) this is a condition with increasing prevalence and (b) diabetics have been shown to actively use the Internet to manage their condition. From January to March 2010 participants were recruited using purposive sampling techniques. A screening instrument was used to determine eligibility. The study consisted of a demographic and computer usage survey, a series of usability tests and an interview. We tested participants (N=22) on two scenarios, each involving tasks that assessed their ability to tag content and search using a tag cloud that included six structured credibility terms (statistics, date, reference, author, testimonial and quotations). MORAE Usability software (version 3.1) was employed to record participants' use of the study environment. The surveys were analyzed using SPSS version 17. Interviews with participants were transcribed, coded and analyzed using thematic text analysis with the aid of NVivo8. Most participants had experience with Internet resources. However, less than one quarter of this sample had seen or used tags or a tag clouds. The ways in which participants used tags to label the content posted in the message forums varied. Some participants were tagging the information for their own subsequent use, whereas others viewed this process from the perspective of others: they tagged the content in ways that they thought other users would find beneficial. Many participants did not use the structured credibility tags when asked to search for credible content. The interviews corroborated these findings by confirming participants were not considering credibility foremost when tagging. Many participants in this study focused on assessing whether the information was relevant to their current circumstances, after which they would proceed to determine its credibility by corroborating with other sources. The use of structured tags to label information may not be a useful way to encourage the use of tagging, or to indicate credibility in this context. Current applications used in the semantic web automate this process. Therefore it may be useful to engage consumers of online content, in particular health-related content, to be more directly involved in the annotation of this content. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  20. [Advances in research and application of beta-tricalcium phosphate, collagen and beta-tricalcium phosphate/collagen composite in bone tissue engineering].

    PubMed

    Han, Xiang-Yong; Fu, Yuan-Fei; Zhang, Fu-Qiang

    2007-02-01

    Bone defects in oral and maxillofacial region was a common problem. To repair the defect, bone grafts including autograft, allograft and artificial bone graft were used in clinic despite of their disadvantages. Nowadays, bone tissue engineering has become a commonly used method to repair bone defect. This paper reviewed the application of beta-TCP, collagen and beta-TCP/collagen composite in bone tissue engineering. It was concluded that beta-TCP/collagen composite was a promising materials in bone tissue engineering.

  1. TIME-TAG mode of STIS observations using the MAMA detectors

    NASA Astrophysics Data System (ADS)

    Sahu, Kailash; Danks, Anthony; Baum, Stefi; Balzano, Vicki; Kraemer, Steve; Kutina, Ray; Sears, William

    1995-04-01

    We summarize the time-tag mode of STIS observations using the MAMA detectors, both in imaging and spectroscopic modes. After a brief outline on the MAMA detector characteristics and the astronomical applications of the time-tag mode, the general philosophy and the details of the data management strategy are described in detail. The GO specifications, and the consequent different modes of data transfer strategy are outlined. Restrictions on maximum data rates, integration times, and BUFFER-TIME requirements are explained. A few cases where the subarray option would be useful are outlined.

  2. Imidazolium tagged acridines: Synthesis, characterization and applications in DNA binding and anti-microbial activities

    NASA Astrophysics Data System (ADS)

    Raju, Gembali; Vishwanath, S.; Prasad, Archana; Patel, Basant K.; Prabusankar, Ganesan

    2016-03-01

    New water soluble 4,5-bis imidazolium tagged acridines have been synthesized and structurally characterized by multinuclear NMR and single crystal X-ray diffraction techniques. The DNA binding and anti-microbial activities of these acridine derivatives were investigated by fluorescence and far-UV circular dichroism studies.

  3. Current radar-responsive tag development activities at Sandia National Laboratories

    NASA Astrophysics Data System (ADS)

    Ormesher, Richard C.; Plummer, Kenneth W.; Wells, Lars M.

    2004-08-01

    Over the past ten years, Sandia has developed RF radar responsive tag systems and supporting technologies for various government agencies and industry partners. RF tags can function as RF transmitters or radar transponders that enable tagging, tracking, and location determination functions. Expertise in tag architecture, microwave and radar design, signal analysis and processing techniques, digital design, modeling and simulation, and testing have been directly applicable to these tag programs. In general, the radar responsive tag designs have emphasized low power, small package size, and the ability to be detected by the radar at long ranges. Recently, there has been an interest in using radar responsive tags for Blue Force tracking and Combat ID (CID). The main reason for this interest is to allow airborne surveillance radars to easily distinguish U.S. assets from those of opposing forces. A Blue Force tracking capability would add materially to situational awareness. Combat ID is also an issue, as evidenced by the fact that approximately one-quarter of all U.S. casualties in the Gulf War took the form of ground troops killed by friendly fire. Because the evolution of warfare in the intervening decade has made asymmetric warfare the norm rather than the exception, swarming engagements in which U.S. forces will be freely intermixed with opposing forces is a situation that must be anticipated. Increasing utilization of precision munitions can be expected to drive fires progressively closer to engaged allied troops at times when visual de-confliction is not an option. In view of these trends, it becomes increasingly important that U.S. ground forces have a widely proliferated all-weather radar responsive tag that communicates to all-weather surveillance. The purpose of this paper is to provide an overview of the recent, current, and future radar responsive research and development activities at Sandia National Laboratories that support both the Blue Force Tracking and Combat ID application. Sandia is a multiprogram laboratory operated by Sandia Corporation, a Lockheed Martin Company for the United States Departments of Energy's National Nuclear Security Administration under contract DE-AC04-94AL85000.

  4. Z-path SAW RFID tag.

    PubMed

    Härmä, Sanna; Plessky, Victor P; Hartmann, Clinton S; Steichen, William

    2008-01-01

    Surface acoustic wave (SAW) radio-frequency identification (RFID) tags are soon expected to be produced in very high volumes. The size and cost of a SAW RFID tag will be key parameters for many applications. Therefore, it is of primary importance to reduce the chip size. In this work, we describe the design principles of a 2.4-GHz SAW RFID tag that is significantly smaller than earlier reported tags. We also present simulated and experimental results. The coded signal should arrive at the reader with a certain delay (typically about 1 micros), i.e., after the reception of environmental echoes. If the tag uses a bidirectional interdigital transducer (IDT), space for the initial delay is needed on both sides of the IDT. In this work, we replace the bidirectional IDT by a unidirectional one. This halves the space required by the initial delay because all the code reflectors must now be placed on the same side of the IDT. We reduce tag size even further by using a Z-path geometry in which the same space in x-direction is used for both the initial delay and the code reflectors. Chip length is thus determined only by the space required by the code reflectors.

  5. Clone tag detection in distributed RFID systems

    PubMed Central

    Kamaludin, Hazalila; Mahdin, Hairulnizam

    2018-01-01

    Although Radio Frequency Identification (RFID) is poised to displace barcodes, security vulnerabilities pose serious challenges for global adoption of the RFID technology. Specifically, RFID tags are prone to basic cloning and counterfeiting security attacks. A successful cloning of the RFID tags in many commercial applications can lead to many serious problems such as financial losses, brand damage, safety and health of the public. With many industries such as pharmaceutical and businesses deploying RFID technology with a variety of products, it is important to tackle RFID tag cloning problem and improve the resistance of the RFID systems. To this end, we propose an approach for detecting cloned RFID tags in RFID systems with high detection accuracy and minimal overhead thus overcoming practical challenges in existing approaches. The proposed approach is based on consistency of dual hash collisions and modified count-min sketch vector. We evaluated the proposed approach through extensive experiments and compared it with existing baseline approaches in terms of execution time and detection accuracy under varying RFID tag cloning ratio. The results of the experiments show that the proposed approach outperforms the baseline approaches in cloned RFID tag detection accuracy. PMID:29565982

  6. Application of the hydroxyl tagging velocimetry to direct-connect supersonic combustor experiment

    NASA Astrophysics Data System (ADS)

    Ye, Jingfeng; Li, Guohua; Shao, Jun; Hu, Zhiyun; Zhao, Xinyan; Song, WenYan

    2017-05-01

    For the purpose of measuring the flow velocity in a scramjet test model, an special designed measurement system was established, including the strong vibration suppression, optical transport consideration, the movable device etc. The interference of the strong vibration to the velocity measurements was avoided by two ICCD cameras capturing the reference tag lines image and moved tag lines image together during an experiment. According to the tag lines image feature, data processing including correlation algorithm, data fitting by a Gauss function were used respectively to extract the positions of the reference tag lines and the moved tag lines. The velocity measurements were carried out at the isolation section and the cavity section. The results showed that the well SNR could be achieved in the H2/air combustion heating flow, but in the kerosene fuel combustion flow, the measurements images might be interfered by the strong OH background from the chemical reaction, and the signal intensity could be reduced due to the tag laser attenuation through the absorption by kerosene vapor. But when the combustor model was run at a low chemical equivalent, the interference could be suppressed to an accepted level.

  7. A distinct DGAT with sn-3 acetyltransferase activity that synthesizes unusual, reduced-viscosity oils in Euonymus and transgenic seeds

    PubMed Central

    Durrett, Timothy P.; McClosky, Daniel D.; Tumaney, Ajay W.; Elzinga, Dezi A.; Ohlrogge, John; Pollard, Mike

    2010-01-01

    Endosperm and embryo tissues from the seeds of Euonymus alatus (Burning Bush) accumulate high levels of 3-acetyl-1,2-diacyl-sn-glycerols (acTAGs) as their major storage lipids. In contrast, the aril tissue surrounding the seed produces long-chain triacylglycerols (lcTAGs) typical of most other organisms. The presence of the sn-3 acetyl group imparts acTAGs with different physical and chemical properties, such as a 30% reduction in viscosity, compared to lcTAGs. Comparative transcriptome analysis of developing endosperm and aril tissues using pyrosequencing technology was performed to isolate the enzyme necessary for the synthesis of acTAGs. An uncharacterized membrane-bound O-acyltransferase (MBOAT) family member was the most abundant acyltransferase in the endosperm but was absent from the aril. Expression of this MBOAT in yeast resulted in the accumulation of acTAGs but not lcTAG; hence, the enzyme was named EaDAcT (Euonymus alatus diacylglycerol acetyltransferase). Yeast microsomes expressing EaDAcT possessed acetyl-CoA diacylglycerol acetyltransferase activity but lacked long-chain acyl-CoA diacylglycerol acyltransferase activity. Expression of EaDAcT under the control of a strong, seed-specific promoter in Arabidopsis resulted in the accumulation of acTAGs, up to 40 mol % of total TAG in the seed oil. These results demonstrate the utility of deep transcriptional profiling with multiple tissues as a gene discovery strategy for low-abundance proteins. They also show that EaDAcT is the acetyltransferase necessary and sufficient for the production of acTAGs in Euonymus seeds, and that this activity can be introduced into the seeds of other plants, allowing the evaluation of these unusual TAGs for biofuel and other applications. PMID:20439724

  8. A distinct DGAT with sn-3 acetyltransferase activity that synthesizes unusual, reduced-viscosity oils in Euonymus and transgenic seeds.

    PubMed

    Durrett, Timothy P; McClosky, Daniel D; Tumaney, Ajay W; Elzinga, Dezi A; Ohlrogge, John; Pollard, Mike

    2010-05-18

    Endosperm and embryo tissues from the seeds of Euonymus alatus (Burning Bush) accumulate high levels of 3-acetyl-1,2-diacyl-sn-glycerols (acTAGs) as their major storage lipids. In contrast, the aril tissue surrounding the seed produces long-chain triacylglycerols (lcTAGs) typical of most other organisms. The presence of the sn-3 acetyl group imparts acTAGs with different physical and chemical properties, such as a 30% reduction in viscosity, compared to lcTAGs. Comparative transcriptome analysis of developing endosperm and aril tissues using pyrosequencing technology was performed to isolate the enzyme necessary for the synthesis of acTAGs. An uncharacterized membrane-bound O-acyltransferase (MBOAT) family member was the most abundant acyltransferase in the endosperm but was absent from the aril. Expression of this MBOAT in yeast resulted in the accumulation of acTAGs but not lcTAG; hence, the enzyme was named EaDAcT (Euonymus alatus diacylglycerol acetyltransferase). Yeast microsomes expressing EaDAcT possessed acetyl-CoA diacylglycerol acetyltransferase activity but lacked long-chain acyl-CoA diacylglycerol acyltransferase activity. Expression of EaDAcT under the control of a strong, seed-specific promoter in Arabidopsis resulted in the accumulation of acTAGs, up to 40 mol % of total TAG in the seed oil. These results demonstrate the utility of deep transcriptional profiling with multiple tissues as a gene discovery strategy for low-abundance proteins. They also show that EaDAcT is the acetyltransferase necessary and sufficient for the production of acTAGs in Euonymus seeds, and that this activity can be introduced into the seeds of other plants, allowing the evaluation of these unusual TAGs for biofuel and other applications.

  9. Accounting for tagging-to-harvest mortality in a Brownie tag-recovery model by incorporating radio-telemetry data.

    PubMed

    Buderman, Frances E; Diefenbach, Duane R; Casalena, Mary Jo; Rosenberry, Christopher S; Wallingford, Bret D

    2014-04-01

    The Brownie tag-recovery model is useful for estimating harvest rates but assumes all tagged individuals survive to the first hunting season; otherwise, mortality between time of tagging and the hunting season will cause the Brownie estimator to be negatively biased. Alternatively, fitting animals with radio transmitters can be used to accurately estimate harvest rate but may be more costly. We developed a joint model to estimate harvest and annual survival rates that combines known-fate data from animals fitted with transmitters to estimate the probability of surviving the period from capture to the first hunting season, and data from reward-tagged animals in a Brownie tag-recovery model. We evaluated bias and precision of the joint estimator, and how to optimally allocate effort between animals fitted with radio transmitters and inexpensive ear tags or leg bands. Tagging-to-harvest survival rates from >20 individuals with radio transmitters combined with 50-100 reward tags resulted in an unbiased and precise estimator of harvest rates. In addition, the joint model can test whether transmitters affect an individual's probability of being harvested. We illustrate application of the model using data from wild turkey, Meleagris gallapavo, to estimate harvest rates, and data from white-tailed deer, Odocoileus virginianus, to evaluate whether the presence of a visible radio transmitter is related to the probability of a deer being harvested. The joint known-fate tag-recovery model eliminates the requirement to capture and mark animals immediately prior to the hunting season to obtain accurate and precise estimates of harvest rate. In addition, the joint model can assess whether marking animals with radio transmitters affects the individual's probability of being harvested, caused by hunter selectivity or changes in a marked animal's behavior.

  10. A Synthesis of Tagging Studies Examining the Behaviour and Survival of Anadromous Salmonids in Marine Environments

    PubMed Central

    Drenner, S. Matthew; Clark, Timothy D.; Whitney, Charlotte K.; Martins, Eduardo G.; Cooke, Steven J.; Hinch, Scott G.

    2012-01-01

    This paper synthesizes tagging studies to highlight the current state of knowledge concerning the behaviour and survival of anadromous salmonids in the marine environment. Scientific literature was reviewed to quantify the number and type of studies that have investigated behaviour and survival of anadromous forms of Pacific salmon (Oncorhynchus spp.), Atlantic salmon (Salmo salar), brown trout (Salmo trutta), steelhead (Oncorhynchus mykiss), and cutthroat trout (Oncorhynchus clarkii). We examined three categories of tags including electronic (e.g. acoustic, radio, archival), passive (e.g. external marks, Carlin, coded wire, passive integrated transponder [PIT]), and biological (e.g. otolith, genetic, scale, parasites). Based on 207 papers, survival rates and behaviour in marine environments were found to be extremely variable spatially and temporally, with some of the most influential factors being temperature, population, physiological state, and fish size. Salmonids at all life stages were consistently found to swim at an average speed of approximately one body length per second, which likely corresponds with the speed at which transport costs are minimal. We found that there is relatively little research conducted on open-ocean migrating salmonids, and some species (e.g. masu [O. masou] and amago [O. rhodurus]) are underrepresented in the literature. The most common forms of tagging used across life stages were various forms of external tags, coded wire tags, and acoustic tags, however, the majority of studies did not measure tagging/handling effects on the fish, tag loss/failure, or tag detection probabilities when estimating survival. Through the interdisciplinary application of existing and novel technologies, future research examining the behaviour and survival of anadromous salmonids could incorporate important drivers such as oceanography, tagging/handling effects, predation, and physiology. PMID:22431962

  11. A novel method for 3D measurement of RFID multi-tag network based on matching vision and wavelet

    NASA Astrophysics Data System (ADS)

    Zhuang, Xiao; Yu, Xiaolei; Zhao, Zhimin; Wang, Donghua; Zhang, Wenjie; Liu, Zhenlu; Lu, Dongsheng; Dong, Dingbang

    2018-07-01

    In the field of radio frequency identification (RFID), the three-dimensional (3D) distribution of RFID multi-tag networks has a significant impact on their reading performance. At the same time, in order to realize the anti-collision of RFID multi-tag networks in practical engineering applications, the 3D distribution of RFID multi-tag networks must be measured. In this paper, a novel method for the 3D measurement of RFID multi-tag networks is proposed. A dual-CCD system (vertical and horizontal cameras) is used to obtain images of RFID multi-tag networks from different angles. Then, the wavelet threshold denoising method is used to remove noise in the obtained images. The template matching method is used to determine the two-dimensional coordinates and vertical coordinate of each tag. The 3D coordinates of each tag are obtained subsequently. Finally, a model of the nonlinear relation between the 3D coordinate distribution of the RFID multi-tag network and the corresponding reading distance is established using the wavelet neural network. The experiment results show that the average prediction relative error is 0.71% and the time cost is 2.17 s. The values of the average prediction relative error and time cost are smaller than those of the particle swarm optimization neural network and genetic algorithm–back propagation neural network. The time cost of the wavelet neural network is about 1% of that of the other two methods. The method proposed in this paper has a smaller relative error. The proposed method can improve the real-time performance of RFID multi-tag networks and the overall dynamic performance of multi-tag networks.

  12. Accounting for tagging-to-harvest mortality in a Brownie tag-recovery model by incorporating radio-telemetry data

    USGS Publications Warehouse

    Buderman, Frances E.; Diefenbach, Duane R.; Casalena, Mary Jo; Rosenberry, Christopher S.; Wallingford, Bret D.

    2014-01-01

    The Brownie tag-recovery model is useful for estimating harvest rates but assumes all tagged individuals survive to the first hunting season; otherwise, mortality between time of tagging and the hunting season will cause the Brownie estimator to be negatively biased. Alternatively, fitting animals with radio transmitters can be used to accurately estimate harvest rate but may be more costly. We developed a joint model to estimate harvest and annual survival rates that combines known-fate data from animals fitted with transmitters to estimate the probability of surviving the period from capture to the first hunting season, and data from reward-tagged animals in a Brownie tag-recovery model. We evaluated bias and precision of the joint estimator, and how to optimally allocate effort between animals fitted with radio transmitters and inexpensive ear tags or leg bands. Tagging-to-harvest survival rates from >20 individuals with radio transmitters combined with 50–100 reward tags resulted in an unbiased and precise estimator of harvest rates. In addition, the joint model can test whether transmitters affect an individual's probability of being harvested. We illustrate application of the model using data from wild turkey, Meleagris gallapavo,to estimate harvest rates, and data from white-tailed deer, Odocoileus virginianus, to evaluate whether the presence of a visible radio transmitter is related to the probability of a deer being harvested. The joint known-fate tag-recovery model eliminates the requirement to capture and mark animals immediately prior to the hunting season to obtain accurate and precise estimates of harvest rate. In addition, the joint model can assess whether marking animals with radio transmitters affects the individual's probability of being harvested, caused by hunter selectivity or changes in a marked animal's behavior.

  13. A synthesis of tagging studies examining the behaviour and survival of anadromous salmonids in marine environments.

    PubMed

    Drenner, S Matthew; Clark, Timothy D; Whitney, Charlotte K; Martins, Eduardo G; Cooke, Steven J; Hinch, Scott G

    2012-01-01

    This paper synthesizes tagging studies to highlight the current state of knowledge concerning the behaviour and survival of anadromous salmonids in the marine environment. Scientific literature was reviewed to quantify the number and type of studies that have investigated behaviour and survival of anadromous forms of Pacific salmon (Oncorhynchus spp.), Atlantic salmon (Salmo salar), brown trout (Salmo trutta), steelhead (Oncorhynchus mykiss), and cutthroat trout (Oncorhynchus clarkii). We examined three categories of tags including electronic (e.g. acoustic, radio, archival), passive (e.g. external marks, Carlin, coded wire, passive integrated transponder [PIT]), and biological (e.g. otolith, genetic, scale, parasites). Based on 207 papers, survival rates and behaviour in marine environments were found to be extremely variable spatially and temporally, with some of the most influential factors being temperature, population, physiological state, and fish size. Salmonids at all life stages were consistently found to swim at an average speed of approximately one body length per second, which likely corresponds with the speed at which transport costs are minimal. We found that there is relatively little research conducted on open-ocean migrating salmonids, and some species (e.g. masu [O. masou] and amago [O. rhodurus]) are underrepresented in the literature. The most common forms of tagging used across life stages were various forms of external tags, coded wire tags, and acoustic tags, however, the majority of studies did not measure tagging/handling effects on the fish, tag loss/failure, or tag detection probabilities when estimating survival. Through the interdisciplinary application of existing and novel technologies, future research examining the behaviour and survival of anadromous salmonids could incorporate important drivers such as oceanography, tagging/handling effects, predation, and physiology.

  14. Comparing effects of transmitters within and among populations: application to swimming performance of juvenile Chinook salmon

    USGS Publications Warehouse

    Perry, Russell W.; Plumb, John M.; Fielding, Scott D.; Adams, Noah S.; Rondorf, Dennis W.

    2013-01-01

    The sensitivity of fish to a transmitter depends on factors such as environmental conditions, fish morphology, life stage, rearing history, and tag design. However, synthesizing general trends across studies is difficult because each study focuses on a particular performance measure, species, life stage, and transmitter model. These differences motivated us to develop simple metrics that allow effects of transmitters to be compared among different species, populations, or studies. First, we describe how multiple regression analysis can be used to quantify the effect of tag burden (transmitter mass relative to fish mass) on measures of physiological performance. Next, we illustrate how the slope and intercept parameters can be used to calculate two summary statistics: θ, which estimates the tag burden threshold above which the performance of tagged fish begins to decline relative to untagged fish; and k, which measures the percentage change in performance per percentage point increase in tag burden. When θ = 0, k provides a single measure of the tag's effect that can be compared among species, populations, or studies. We apply this analysis to two different experiments that measure the critical swimming speed (U crit) of tagged juvenile Chinook Salmon Oncorhynchus tshawytscha. In both experiments, U crit declined as tag burden increased, but we found no significant threshold in swimming performance. Estimates of θ ranged from −0.6% to 2.1% among six unique treatment groups, indicating that swimming performance began to decline at a relatively low tag burden. Estimates of k revealed that U crit of tagged fish declined by −2.68% to −4.86% for each 1% increase in tag burden. Both θ and k varied with the tag's antenna configuration, tag implantation method, and posttagging recovery time. Our analytical approach can be used to gain insights across populations to better understand factors affecting the ability of fish to carry a transmitter.

  15. Phospholipase Dζ Enhances Diacylglycerol Flux into Triacylglycerol

    DOE PAGES

    Yang, Wenyu; Wang, Geliang; Li, Jia; ...

    2017-03-21

    Plant seeds are the primary source of triacylglycerols (TAG) for food, feed, fuel, and industrial applications. As TAG is produced from diacylglycerol (DAG), successful engineering strategies to enhance TAG levels have focused on the conversion of DAG to TAG. However, the production of TAG can be limited by flux through the enzymatic reactions that supply DAG. In this study, two Arabidopsis phospholipase Dζ genes (AtPLDζ 1 and AtPLDζ 2) were coexpressed in Camelina sativa to test whether the conversion of phosphatidylcholine to DAG impacts TAG levels in seeds. The resulting transgenic plants produced 2% to 3% more TAG as amore » component of total seed biomass and had increased 18:3 and 20:1 fatty acid levels relative to wild type. Increased DAG and decreased PC levels were examined through the kinetics of lipid assembly by [ 14C]acetate and [ 14C]glycerol incorporation into glycerolipids. [ 14C]acetate was rapidly incorporated into TAG in both wild-type and overexpression lines, indicating a significant flux of nascent and elongated acyl-CoAs into the sn-3 position of TAG. Stereochemical analysis revealed that newly synthesized fatty acids were preferentially incorporated into the sn-2 position of PC, but the sn-1 position of de novo DAG and indicated similar rates of nascent acyl groups into the Kennedy pathway and acyl editing. [ 14C]glycerol studies demonstrated PC-derived DAG is the major source of DAG for TAG synthesis in both tissues. The results emphasize that the interconversions of DAG and PC pools can impact oil production and composition.« less

  16. Phospholipase Dζ Enhances Diacylglycerol Flux into Triacylglycerol

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yang, Wenyu; Wang, Geliang; Li, Jia

    Plant seeds are the primary source of triacylglycerols (TAG) for food, feed, fuel, and industrial applications. As TAG is produced from diacylglycerol (DAG), successful engineering strategies to enhance TAG levels have focused on the conversion of DAG to TAG. However, the production of TAG can be limited by flux through the enzymatic reactions that supply DAG. In this study, two Arabidopsis phospholipase Dζ genes (AtPLDζ 1 and AtPLDζ 2) were coexpressed in Camelina sativa to test whether the conversion of phosphatidylcholine to DAG impacts TAG levels in seeds. The resulting transgenic plants produced 2% to 3% more TAG as amore » component of total seed biomass and had increased 18:3 and 20:1 fatty acid levels relative to wild type. Increased DAG and decreased PC levels were examined through the kinetics of lipid assembly by [ 14C]acetate and [ 14C]glycerol incorporation into glycerolipids. [ 14C]acetate was rapidly incorporated into TAG in both wild-type and overexpression lines, indicating a significant flux of nascent and elongated acyl-CoAs into the sn-3 position of TAG. Stereochemical analysis revealed that newly synthesized fatty acids were preferentially incorporated into the sn-2 position of PC, but the sn-1 position of de novo DAG and indicated similar rates of nascent acyl groups into the Kennedy pathway and acyl editing. [ 14C]glycerol studies demonstrated PC-derived DAG is the major source of DAG for TAG synthesis in both tissues. The results emphasize that the interconversions of DAG and PC pools can impact oil production and composition.« less

  17. Molecular tagging techniques and their applications to the study of complex thermal flow phenomena

    NASA Astrophysics Data System (ADS)

    Chen, Fang; Li, Haixing; Hu, Hui

    2015-08-01

    This review article reports the recent progress in the development of a new group of molecule-based flow diagnostic techniques, which include molecular tagging velocimetry (MTV) and molecular tagging thermometry (MTT), for both qualitative flow visualization of thermally induced flow structures and quantitative whole-field measurements of flow velocity and temperature distributions. The MTV and MTT techniques can also be easily combined to result in a so-called molecular tagging velocimetry and thermometry (MTV&T) technique, which is capble of achieving simultaneous measurements of flow velocity and temperature distribution in fluid flows. Instead of using tiny particles, the molecular tagging techniques (MTV, MTT, and MTV&T) use phosphorescent molecules, which can be turned into long-lasting glowing marks upon excitation by photons of appropriate wavelength, as the tracers for the flow velocity and temperature measurements. The unique attraction and implementation of the molecular tagging techniques are demonstrated by three application examples, which include: (1) to quantify the unsteady heat transfer process from a heated cylinder to the surrounding fluid flow in order to examine the thermal effects on the wake instabilities behind the heated cylinder operating in mixed and forced heat convection regimes, (2) to reveal the time evolution of unsteady heat transfer and phase changing process inside micro-sized, icing water droplets in order to elucidate the underlying physics pertinent to aircraft icing phenomena, and (3) to achieve simultaneous droplet size, velocity and temperature measurements of "in-flight" droplets to characterize the dynamic and thermodynamic behaviors of flying droplets in spray flows.

  18. Optimal use of tandem biotin and V5 tags in ChIP assays

    PubMed Central

    Kolodziej, Katarzyna E; Pourfarzad, Farzin; de Boer, Ernie; Krpic, Sanja; Grosveld, Frank; Strouboulis, John

    2009-01-01

    Background Chromatin immunoprecipitation (ChIP) assays coupled to genome arrays (Chip-on-chip) or massive parallel sequencing (ChIP-seq) lead to the genome wide identification of binding sites of chromatin associated proteins. However, the highly variable quality of antibodies and the availability of epitopes in crosslinked chromatin can compromise genomic ChIP outcomes. Epitope tags have often been used as more reliable alternatives. In addition, we have employed protein in vivo biotinylation tagging as a very high affinity alternative to antibodies. In this paper we describe the optimization of biotinylation tagging for ChIP and its coupling to a known epitope tag in providing a reliable and efficient alternative to antibodies. Results Using the biotin tagged erythroid transcription factor GATA-1 as example, we describe several optimization steps for the application of the high affinity biotin streptavidin system in ChIP. We find that the omission of SDS during sonication, the use of fish skin gelatin as blocking agent and choice of streptavidin beads can lead to significantly improved ChIP enrichments and lower background compared to antibodies. We also show that the V5 epitope tag performs equally well under the conditions worked out for streptavidin ChIP and that it may suffer less from the effects of formaldehyde crosslinking. Conclusion The combined use of the very high affinity biotin tag with the less sensitive to crosslinking V5 tag provides for a flexible ChIP platform with potential implications in ChIP sequencing outcomes. PMID:19196479

  19. Molecular cloning, expression, purification and osteoblasts proliferation activity of sika deer thymosin beta10.

    PubMed

    Wang, J; Liu, M; Bai, X; Zhang, H; Xu, Z; Zhao, D; Zhao, Y

    2017-12-01

    Thymosin beta 10 (Tβ10) is a member of the β-thymosin family. As an actin-binding peptide, thymosin β10 is involved in many important biological activities. Transcriptome sequencing results suggest that Tβ10 may play important roles in the growth of deer antler. In this study, Tβ10 cDNA was isolated from sika deer, and complete open reading frame consisting of 129 nucleotides was obtained by PCR amplification. The predicted peptide was 42 amino acids in length. The sdTβ10 cDNA was cloned and expressed in Escherichia coli resulting in a 6 kDa recombinant-His tagged protein. The recombinant, non-glycosylated peptide was overexpressed in a soluble form and purified by immobilized metal ion affinity chromatography. Functional studies revealed that recombinant Tβ10 stimulated osteoblasts proliferation. This study provides the first evidence that recombinant sika deer Tβ10 promotes proliferation in an osteoblasts cell model. Copyright© by the Polish Academy of Sciences.

  20. Interleukin 1 beta gene and risk of schizophrenia: detailed case-control and family-based studies and an updated meta-analysis.

    PubMed

    Shibuya, Masako; Watanabe, Yuichiro; Nunokawa, Ayako; Egawa, Jun; Kaneko, Naoshi; Igeta, Hirofumi; Someya, Toshiyuki

    2014-01-01

    Interleukin-1 beta (IL-1β) has been implicated in the pathophysiology of schizophrenia. To assess whether the IL1B gene confers increased susceptibility to schizophrenia, we conducted case-control and family-based studies and an updated meta-analysis. We tested the association between IL1B and schizophrenia in 1229 case-control and 112 trio samples using 12 markers, including common tagging single nucleotide variations (SNVs) and a rare non-synonymous variation detected by resequencing the coding regions. We also performed a meta-analysis of rs16944 using a total of 8724 case-control and 201 trio samples from 16 independent populations. We found no significant associations between any of the 12 SNVs examined and schizophrenia in either case-control or trio samples. Moreover, our meta-analysis results showed no significant association between the common SNV, rs16944, and schizophrenia. The present study does not support a role for IL1B in schizophrenia susceptibility.

  1. Synthesis of High-Load, Hybrid Silica-Immobilized Heterocyclic Benzyl Phosphate (Si–OHBP) and Triazolyl Phosphate (Si–OHTP) Alkylating Reagents

    PubMed Central

    2016-01-01

    The development of new ROMP-derived silica-immobilized heterocyclic phosphate reagents and their application in purification-free protocols is reported. Grafting of norbornenyl norbornenyl-functionalized (Nb-tagged) silica particles with functionalized Nb-tagged heterocyclic phosphate monomers efficiently yield high-load, hybrid silica-immobilized oligomeric heterobenzyl phosphates (Si–OHBP) and heterotriazolyl phosphates (Si–OHTP) as efficient alkylation agents. Applications of these reagents for the diversification of N-, O-, and S-nucleophilic species, for efficient heterobenzylation and hetero(triazolyl)methylation have been validated. PMID:27300761

  2. Associated Particle Tagging (APT) in Magnetic Spectrometers

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Jordan, David V.; Baciak, James E.; Stave, Sean C.

    2012-10-16

    Summary In Brief The Associated Particle Tagging (APT) project, a collaboration of Pacific Northwest National Laboratory (PNNL), Idaho National Laboratory (INL) and the Idaho State University (ISU)/Idaho Accelerator Center (IAC), has completed an exploratory study to assess the role of magnetic spectrometers as the linchpin technology in next-generation tagged-neutron and tagged-photon active interrogation (AI). The computational study considered two principle concepts: (1) the application of a solenoidal alpha-particle spectrometer to a next-generation, large-emittance neutron generator for use in the associated particle imaging technique, and (2) the application of tagged photon beams to the detection of fissile material via active interrogation.more » In both cases, a magnetic spectrometer momentum-analyzes charged particles (in the neutron case, alpha particles accompanying neutron generation in the D-T reaction; in the tagged photon case, post-bremsstrahlung electrons) to define kinematic properties of the relevant neutral interrogation probe particle (i.e. neutron or photon). The main conclusions of the study can be briefly summarized as follows: Neutron generator: • For the solenoidal spectrometer concept, magnetic field strengths of order 1 Tesla or greater are required to keep the transverse size of the spectrometer smaller than 1 meter. The notional magnetic spectrometer design evaluated in this feasibility study uses a 5-T magnetic field and a borehole radius of 18 cm. • The design shows a potential for 4.5 Sr tagged neutron solid angle, a factor of 4.5 larger than achievable with current API neutron-generator designs. • The potential angular resolution for such a tagged neutron beam can be less than 0.5o for modest Si-detector position resolution (3 mm). Further improvement in angular resolution can be made by using Si-detectors with better position resolution. • The report documents several features of a notional generator design incorporating the alpha-particle spectrometer concept, and outlines challenges involved in the magnetic field design. Tagged photon interrogation: • We investigated a method for discriminating fissile from benign cargo-material response to an energy-tagged photon beam. The method relies upon coincident detection of the tagged photon and a photoneutron or photofission neutron produced in the target material. The method exploits differences in the shape of the neutron production cross section as a function of incident photon energy in order to discriminate photofission yield from photoneutrons emitted by non-fissile materials. Computational tests of the interrogation method as applied to material composition assay of a simple, multi-layer target suggest that the tagged-photon information facilitates precise (order 1% thickness uncertainty) reconstruction of the constituent thicknesses of fissile (uranium) and high-Z (Pb) constituents of the test targets in a few minutes of photon-beam exposure. We assumed an 18-MeV endpoint tagged photon beam for these simulations. • The report addresses several candidate design and data analysis issues for beamline infrastructure required to produce a tagged photon beam in a notional AI-dedicated facility, including the accelerator and tagging spectrometer.« less

  3. Novel cell-based assay for detection of thyroid receptor beta-interacting environmental contaminants.

    PubMed

    Stavreva, Diana A; Varticovski, Lyuba; Levkova, Ludmila; George, Anuja A; Davis, Luke; Pegoraro, Gianluca; Blazer, Vicki; Iwanowicz, Luke; Hager, Gordon L

    2016-08-10

    Even though the presence of endocrine disrupting chemicals (EDCs) with thyroid hormone (TH)-like activities in the environment is a major health concern, the methods for their efficient detection and monitoring are still limited. Here we describe a novel cell assay, based on the translocation of a green fluorescent protein (GFP)-tagged chimeric molecule of glucocorticoid receptor (GR) and the thyroid receptor beta (TRβ) from the cytoplasm to the nucleus in the presence of TR ligands. Unlike the constitutively nuclear TRβ, this GFP-GR-TRβ chimera is cytoplasmic in the absence of hormone while translocating to the nucleus in a time- and concentration-dependent manner upon stimulation with triiodothyronine (T3) and thyroid hormone analogue, TRIAC, while the reverse triiodothyronine (3,3',5'-triiodothyronine, or rT3) was inactive. Moreover, GFP-GR-TRβ chimera does not show any cross-reactivity with the GR-activating hormones, thus providing a clean system for the screening of TR beta-interacting EDCs. Using this assay, we demonstrated that Bisphenol A (BPA) and 3,3',5,5'-Tetrabromobisphenol (TBBPA) induced GFP-GR-TRβ translocation at micro molar concentrations. We screened over 100 concentrated water samples from different geographic locations in the United States and detected a low, but reproducible contamination in 53% of the samples. This system provides a novel high-throughput approach for screening for endocrine disrupting chemicals (EDCs) interacting with TR beta. Published by Elsevier Ireland Ltd.

  4. Novel cell-based assay for detection of thyroid receptor beta-interacting environmental contaminants

    USGS Publications Warehouse

    Stavreva, Diana A.; Varticovski, Lyuba; Levkova, Ludmila; George, Anuja A.; Davis, Luke; Pegoraro, Gianluca; Blazer, Vicki S.; Iwanowicz, Luke R.; Hager, Gordon L.

    2016-01-01

    Even though the presence of endocrine disrupting chemicals (EDCs) with thyroid hormone (TH)-like activities in the environment is a major health concern, the methods for their efficient detection and monitoring are still limited. Here we describe a novel cell assay, based on the translocation of a green fluorescent protein (GFP)—tagged chimeric molecule of glucocorticoid receptor (GR) and the thyroid receptor beta (TRβ) from the cytoplasm to the nucleus in the presence of TR ligands. Unlike the constitutively nuclear TRβ, this GFP-GR-TRβ chimera is cytoplasmic in the absence of hormone while translocating to the nucleus in a time- and concentration-dependent manner upon stimulation with triiodothyronine (T3) and thyroid hormone analogue, TRIAC, while the reverse triiodothyronine (3,3′,5′-triiodothyronine, or rT3) was inactive. Moreover, GFP-GR-TRβ chimera does not show any cross-reactivity with the GR-activating hormones, thus providing a clean system for the screening of TR beta-interacting EDCs. Using this assay, we demonstrated that Bisphenol A (BPA) and 3,3′,5,5′-Tetrabromobisphenol (TBBPA) induced GFP-GR-TRβ translocation at micro molar concentrations. We screened over 100 concentrated water samples from different geographic locations in the United States and detected a low, but reproducible contamination in 53% of the samples. This system provides a novel high-throughput approach for screening for endocrine disrupting chemicals (EDCs) interacting with TR beta.

  5. Neuroligin trafficking deficiencies arising from mutations in the alpha/beta-hydrolase fold protein family.

    PubMed

    De Jaco, Antonella; Lin, Michael Z; Dubi, Noga; Comoletti, Davide; Miller, Meghan T; Camp, Shelley; Ellisman, Mark; Butko, Margaret T; Tsien, Roger Y; Taylor, Palmer

    2010-09-10

    Despite great functional diversity, characterization of the alpha/beta-hydrolase fold proteins that encompass a superfamily of hydrolases, heterophilic adhesion proteins, and chaperone domains reveals a common structural motif. By incorporating the R451C mutation found in neuroligin (NLGN) and associated with autism and the thyroglobulin G2320R (G221R in NLGN) mutation responsible for congenital hypothyroidism into NLGN3, we show that mutations in the alpha/beta-hydrolase fold domain influence folding and biosynthetic processing of neuroligin3 as determined by in vitro susceptibility to proteases, glycosylation processing, turnover, and processing rates. We also show altered interactions of the mutant proteins with chaperones in the endoplasmic reticulum and arrest of transport along the secretory pathway with diversion to the proteasome. Time-controlled expression of a fluorescently tagged neuroligin in hippocampal neurons shows that these mutations compromise neuronal trafficking of the protein, with the R451C mutation reducing and the G221R mutation virtually abolishing the export of NLGN3 from the soma to the dendritic spines. Although the R451C mutation causes a local folding defect, the G221R mutation appears responsible for more global misfolding of the protein, reflecting their sequence positions in the structure of the protein. Our results suggest that disease-related mutations in the alpha/beta-hydrolase fold domain share common trafficking deficiencies yet lead to discrete congenital disorders of differing severity in the endocrine and nervous systems.

  6. Novel cell-based assay for detection of thyroid receptor beta-interacting environmental contaminants

    PubMed Central

    Stavreva, Diana A.; Varticovski, Lyuba; Levkova, Ludmila; George, Anuja A.; Davis, Luke; Pegoraro, Gianluca; Blazer, Vicki; Iwanowicz, Luke; Hager, Gordon L.

    2016-01-01

    Even though the presence of endocrine disrupting chemicals (EDCs) with thyroid hormone (TH)-like activities in the environment is a major health concern, the methods for their efficient detection and monitoring are still limited. Here we describe a novel cell assay, based on the translocation of a green fluorescent protein (GFP) - tagged chimeric molecule of glucocorticoid receptor (GR) and the thyroid receptor beta (TRβ) from the cytoplasm to the nucleus in the presence of TR ligands. Unlike the constitutively nuclear TRβ, this GFP-GR-TRβ chimera is cytoplasmic in the absence of hormone while translocating to the nucleus in a time- and concentration-dependent manner upon stimulation with triiodothyronine (T3) and thyroid hormone analogue, TRIAC, while the reverse triiodothyronine (3,3′,5′-triiodothyronine, or rT3) was inactive. Moreover, GFP-GR-TRβ chimera does not show any cross-reactivity with the GR-activating hormones, thus providing a clean system for the screening of TR beta -interacting EDCs. Using this assay, we demonstrated that Bisphenol A (BPA) and 3,3′,5,5′-Tetrabromobisphenol (TBBPA) induced GFP-GR-TRβ translocation at micro molar concentrations. We screened over 100 concentrated water samples from different geographic locations in the United States and detected a low, but reproducible contamination in 53 % of the samples. This system provides a novel high-throughput approach for screening for endocrine disrupting chemicals (EDCs) interacting with TR beta. PMID:27528272

  7. Enhanced UHF RFID tags for drug tracing.

    PubMed

    Catarinucci, Luca; Colella, Riccardo; De Blasi, Mario; Patrono, Luigi; Tarricone, Luciano

    2012-12-01

    Radio Frequency Identification (RFID) technology is playing a crucial role for item-level tracing systems in healthcare scenarios. The pharmaceutical supply chain is a fascinating application context, where RFID can guarantee transparency in the drug flow, supporting both suppliers and consumers against the growing counterfeiting problem. In such a context, the choice of the most adequate RFID tag, in terms of shape, frequency, size and reading range, is crucial. The potential presence of items containing materials hostile to the electromagnetic propagation exasperates the problem. In addition, the peculiarities of the different RFID-based checkpoints make even more stringent the requirements for the tag. In this work, the performance of several commercial UHF RFID tags in each step of the pharmaceutical supply chain has been evaluated, confirming the expected criticality. On such basis, a guideline for the electromagnetic design of new high-performance tags capable to overcome such criticalities has been defined. Finally, driven by such guidelines, a new enhanced tag has been designed, realized and tested. Due to patent pending issues, the antenna shape is not shown. Nevertheless, the optimal obtained results do not lose their validity. Indeed, on the one hand they demonstrate that high performance item level tracing systems can actually be implemented also in critical operating conditions. On the other hand, they encourage the tag designer to follow the identified guidelines so to realize enhanced UHF tags.

  8. Miniaturised wireless smart tag for optical chemical analysis applications.

    PubMed

    Steinberg, Matthew D; Kassal, Petar; Tkalčec, Biserka; Murković Steinberg, Ivana

    2014-01-01

    A novel miniaturised photometer has been developed as an ultra-portable and mobile analytical chemical instrument. The low-cost photometer presents a paradigm shift in mobile chemical sensor instrumentation because it is built around a contactless smart card format. The photometer tag is based on the radio-frequency identification (RFID) smart card system, which provides short-range wireless data and power transfer between the photometer and a proximal reader, and which allows the reader to also energise the photometer by near field electromagnetic induction. RFID is set to become a key enabling technology of the Internet-of-Things (IoT), hence devices such as the photometer described here will enable numerous mobile, wearable and vanguard chemical sensing applications in the emerging connected world. In the work presented here, we demonstrate the characterisation of a low-power RFID wireless sensor tag with an LED/photodiode-based photometric input. The performance of the wireless photometer has been tested through two different model analytical applications. The first is photometry in solution, where colour intensity as a function of dye concentration was measured. The second is an ion-selective optode system in which potassium ion concentrations were determined by using previously well characterised bulk optode membranes. The analytical performance of the wireless photometer smart tag is clearly demonstrated by these optical absorption-based analytical experiments, with excellent data agreement to a reference laboratory instrument. © 2013 Elsevier B.V. All rights reserved.

  9. Applications suitable for unmanned and autonomous missions utilizing the Tactical Amphibious Ground Support (TAGS) platform

    NASA Astrophysics Data System (ADS)

    Lewis, Paul J.; Torrie, Mitchel R.; Omilon, Paul M.

    2004-09-01

    The value of unmanned vehicles is directly related to the applications to which it can be successfully applied. Many applications exist and have been identified as suitable for unmanned vehicles, especially those involving dull, dirty, difficult, and dangerous tasks. This paper will highlight applications, missions, and capabilities that have been demonstrated on the TAGS platform to date as well as future application and mission considerations. When evaluating real world applications for this type of vehicle, one must take into account and balance the complexity inherent to the control and safeguarding requirements of a large autonomous ground vehicle with the simplicity required for commercial or military field use. In addition, suitability for a particular application may be limited by the size, weight, fuel consumption, reliability, terrain crossing capability, and other abilities of a vehicle and the intelligent software system and sensors commanding it.

  10. Structural and functional determinants of conserved lipid interaction domains of inward rectifying Kir6.2 channels.

    PubMed

    Cukras, Catherine A; Jeliazkova, Iana; Nichols, Colin G

    2002-06-01

    All members of the inward rectifiier K(+) (Kir) channel family are activated by phosphoinositides and other amphiphilic lipids. To further elucidate the mechanistic basis, we examined the membrane association of Kir6.2 fragments of K(ATP) channels, and the effects of site-directed mutations of these fragments and full-length Kir6.2 on membrane association and K(ATP) channel activity, respectively. GFP-tagged Kir6.2 COOH terminus and GFP-tagged pleckstrin homology domain from phospholipase C delta1 both associate with isolated membranes, and association of each is specifically reduced by muscarinic m1 receptor-mediated phospholipid depletion. Kir COOH termini are predicted to contain multiple beta-strands and a conserved alpha-helix (residues approximately 306-311 in Kir6.2). Systematic mutagenesis of D307-F315 reveals a critical role of E308, I309, W311 and F315, consistent with residues lying on one side of a alpha-helix. Together with systematic mutation of conserved charges, the results define critical determinants of a conserved domain that underlies phospholipid interaction in Kir channels.

  11. Identification and characterization of poorly differentiated invasive carcinomas in a mouse model of pancreatic neuroendocrine tumorigenesis.

    PubMed

    Hunter, Karen E; Quick, Marsha L; Sadanandam, Anguraj; Hanahan, Douglas; Joyce, Johanna A

    2013-01-01

    Pancreatic neuroendocrine tumors (PanNETs) are a relatively rare but clinically challenging tumor type. In particular, high grade, poorly-differentiated PanNETs have the worst patient prognosis, and the underlying mechanisms of disease are poorly understood. In this study we have identified and characterized a previously undescribed class of poorly differentiated PanNETs in the RIP1-Tag2 mouse model. We found that while the majority of tumors in the RIP1-Tag2 model are well-differentiated insulinomas, a subset of tumors had lost multiple markers of beta-cell differentiation and were highly invasive, leading us to term them poorly differentiated invasive carcinomas (PDICs). In addition, we found that these tumors exhibited a high mitotic index, resembling poorly differentiated (PD)-PanNETs in human patients. Interestingly, we identified expression of Id1, an inhibitor of DNA binding gene, and a regulator of differentiation, specifically in PDIC tumor cells by histological analysis. The identification of PDICs in this mouse model provides a unique opportunity to study the pathology and molecular characteristics of PD-PanNETs.

  12. Backward Channel Protection Based on Randomized Tree-Walking Algorithm and Its Analysis for Securing RFID Tag Information and Privacy

    NASA Astrophysics Data System (ADS)

    Choi, Wonjoon; Yoon, Myungchul; Roh, Byeong-Hee

    Eavesdropping on backward channels in RFID environments may cause severe privacy problems because it means the exposure of personal information related to tags that each person has. However, most existing RFID tag security schemes are focused on the forward channel protections. In this paper, we propose a simple but effective method to solve the backward channel eavesdropping problem based on Randomized-tree walking algorithm for securing tag ID information and privacy in RFID-based applications. In order to show the efficiency of the proposed scheme, we derive two performance models for the cases when CRC is used and not used. It is shown that the proposed method can lower the probability of eavesdropping on backward channels near to ‘0.’

  13. A fluorogenic probe for SNAP-tagged plasma membrane proteins based on the solvatochromic molecule Nile Red.

    PubMed

    Prifti, Efthymia; Reymond, Luc; Umebayashi, Miwa; Hovius, Ruud; Riezman, Howard; Johnsson, Kai

    2014-03-21

    A fluorogenic probe for plasma membrane proteins based on the dye Nile Red and SNAP-tag is introduced. It takes advantage of Nile Red, a solvatochromic molecule highly fluorescent in an apolar environment, such as cellular membranes, but almost dark in a polar aqueous environment. The probe possesses a tuned affinity for membranes allowing its Nile Red moiety to insert into the lipid bilayer of the plasma membrane, becoming fluorescent, only after its conjugation to a SNAP-tagged plasma membrane protein. The fluorogenic character of the probe was demonstrated for different SNAP-tag fusion proteins, including the human insulin receptor. This work introduces a new approach for generating a powerful turn-on probe for "no-wash" labeling of plasma membrane proteins with numerous applications in bioimaging.

  14. Size-biased distributions in the generalized beta distribution family, with applications to forestry

    Treesearch

    Mark J. Ducey; Jeffrey H. Gove

    2015-01-01

    Size-biased distributions arise in many forestry applications, as well as other environmental, econometric, and biomedical sampling problems. We examine the size-biased versions of the generalized beta of the first kind, generalized beta of the second kind and generalized gamma distributions. These distributions include, as special cases, the Dagum (Burr Type III),...

  15. Possible association between Interleukin-1beta gene and schizophrenia in a Japanese population

    PubMed Central

    2011-01-01

    Background Several lines of evidence have implicated the pro-inflammatory cytokine interleukin-1beta (IL-1β) in the etiology of schizophrenia. Although a number of genetic association studies have been reported, very few have systematically examined gene-wide tagging polymorphisms. Methods A total of 533 patients with schizophrenia (302 males: mean age ± standard deviation 43.4 ± 13.0 years; 233 females; mean age 44.8 ± 15.3 years) and 1136 healthy controls (388 males: mean age 44.6 ± 17.3 years; 748 females; 46.3 ± 15.6 years) were recruited for this study. All subjects were biologically unrelated Japanese individuals. Five tagging polymorphisms of IL-1β gene (rs2853550, rs1143634, rs1143633, rs1143630, rs16944) were examined for association with schizophrenia. Results Significant difference in allele distribution was found between patients with schizophrenia and controls for rs1143633 (P = 0.0089). When the analysis was performed separately in each gender, significant difference between patients and controls in allele distribution of rs1143633 was observed in females (P = 0.0073). A trend towards association was also found between rs16944 and female patients with schizophrenia (P = 0.032). Conclusions The present study shows the first evidence that the IL-1β gene polymorphism rs1143633 is associated with schizophrenia susceptibility in a Japanese population. The results suggest the possibility that the influence of IL-1β gene variations on susceptibility to schizophrenia may be greater in females than in males. Findings of the present study provide further support for the role of IL-1β in the etiology of schizophrenia. PMID:21843369

  16. Tagging polyketides/non-ribosomal peptides with a clickable functionality and applications

    PubMed Central

    Zhu, Xuejun; Zhang, Wenjun

    2015-01-01

    Bioorthogonal chemistry has recently emerged to be one of the most powerful tools in drug discovery and chemical biology. The exploration of it has successfully advanced the field of natural product research. In this Perspective, we survey current strategies for the installation of chemical handles into the molecular scaffolds of several major classes of natural products, including polyketides (PKs), non-ribosomal peptides (NRPs), and their hybrids. By tagging these natural products with chemical handles and coupling them with subsequent bioorthogonal reactions, researchers have visualized and studied the mode of action of natural products, as well as synthesized derivatives with better pharmaceutical properties. We conclude this Perspective by considering two questions: is there a general way to synthesize tagged PKs/NRPs? Does natural product labeling have a broader impact in the field of natural product research beyond current known applications? PMID:25815285

  17. Adaptation of G-TAG Software for Validating Touch-and-Go Comet Surface Sampling Design Methodology

    NASA Technical Reports Server (NTRS)

    Mandic, Milan; Acikmese, Behcet; Blackmore, Lars

    2011-01-01

    The G-TAG software tool was developed under the R&TD on Integrated Autonomous Guidance, Navigation, and Control for Comet Sample Return, and represents a novel, multi-body dynamics simulation software tool for studying TAG sampling. The G-TAG multi-body simulation tool provides a simulation environment in which a Touch-and-Go (TAG) sampling event can be extensively tested. TAG sampling requires the spacecraft to descend to the surface, contact the surface with a sampling collection device, and then to ascend to a safe altitude. The TAG event lasts only a few seconds but is mission-critical with potentially high risk. Consequently, there is a need for the TAG event to be well characterized and studied by simulation and analysis in order for the proposal teams to converge on a reliable spacecraft design. This adaptation of the G-TAG tool was developed to support the Comet Odyssey proposal effort, and is specifically focused to address comet sample return missions. In this application, the spacecraft descends to and samples from the surface of a comet. Performance of the spacecraft during TAG is assessed based on survivability and sample collection performance. For the adaptation of the G-TAG simulation tool to comet scenarios, models are developed that accurately describe the properties of the spacecraft, approach trajectories, and descent velocities, as well as the models of the external forces and torques acting on the spacecraft. The adapted models of the spacecraft, descent profiles, and external sampling forces/torques were more sophisticated and customized for comets than those available in the basic G-TAG simulation tool. Scenarios implemented include the study of variations in requirements, spacecraft design (size, locations, etc. of the spacecraft components), and the environment (surface properties, slope, disturbances, etc.). The simulations, along with their visual representations using G-View, contributed to the Comet Odyssey New Frontiers proposal effort by indicating problems and/or benefits of different approaches and designs.

  18. Miniaturizing RFID for magnamosis.

    PubMed

    Jiang, Hao; Chen, Shijie; Kish, Shad; Loh, Lokkee; Zhang, Junmin; Zhang, Xiaorong; Kwiat, Dillon; Harrison, Michael; Roy, Shuvo

    2014-01-01

    Anastomosis is a common surgical procedure using staples or sutures in an open or laparoscopic surgery. A more effective and much less invasive alternative is to apply the mechanical pressure on the tissue over a few days [1]. Since the pressure is produced by the attractive force between two permanent magnets, the procedure is called magnamosis[1]. To ensure the two magnets are perfectly aligned during the surgery, a miniaturized batteryless Radio Frequency IDentification (RFID) tag is developed to wirelessly telemeter the status of a pressure sensitive mechanical switch. Using the multi-layer circular spiral coil design, the diameter of the RFID tag is shrunk to 10, 15, 19 and 27 mm to support the magnamosis for children as well as adults. With the impedance matching network, the operating distance of these four RFID tags are longer than 10 cm in a 20 × 22 cm(2) area, even when the tag's normal direction is 45° off the antenna's normal direction. Measurement results also indicate that there is no noticeable degradation on the operating distance when the tag is immersed in saline or placed next to the rare-earth magnet. The miniaturized RFID tag presented in this paper is able to support the magnamosis and other medical applications that require the miniaturized RFID tag.

  19. Crystal structure of a polyhistidine-tagged recombinant catalytic subunit of cAMP-dependent protein kinase complexed with the peptide inhibitor PKI(5-24) and adenosine.

    PubMed

    Narayana, N; Cox, S; Shaltiel, S; Taylor, S S; Xuong, N

    1997-04-15

    The crystal structure of the hexahistidine-tagged mouse recombinant catalytic subunit (H6-rC) of cAMP-dependent protein kinase (cAPK), complexed with a 20-residue peptide inhibitor from the heat-stable protein kinase inhibitor PKI(5-24) and adenosine, was determined at 2.2 A resolution. Novel crystallization conditions were required to grow the ternary complex crystals. The structure was refined to a final crystallographic R-factor of 18.2% with good stereochemical parameters. The "active" enzyme adopts a "closed" conformation as found in rC:PKI(5-24) [Knighton et al. (1991a,b) Science 253, 407-414, 414-420] and packs in a similar manner with the peptide providing a major contact surface. This structure clearly defines the subsites of the unique nucleotide binding site found in the protein kinase family. The adenosine occupies a mostly hydrophobic pocket at the base of the cleft between the two lobes and is completely buried. The missing triphosphate moiety of ATP is filled with a water molecule (Wtr 415) which replaces the gamma-phosphate of ATP. The glycine-rich loop between beta1 and beta2 helps to anchor the phosphates while the ribose ring is buried beneath beta-strand 2. Another ordered water molecule (Wtr 375) is pentacoordinated with polar atoms from adenosine, Leu 49 in beta-strand 1, Glu 127 in the linker strand between the two lobes, Tyr 330, and a third water molecule, Wtr 359. The conserved nucleotide fold can be defined as a lid comprised of beta-strand 1, the glycine-rich loop, and beta-strand 2. The adenine ring is buried beneath beta-strand 1 and the linker strand (120-127) that joins the small and large lobes. The C-terminal tail containing Tyr 330, a segment that lies outside the conserved core, covers this fold and anchors it in a closed conformation. The main-chain atoms of the flexible glycine-rich loop (residues 50-55) in the ATP binding domain have a mean B-factor of 41.4 A2. This loop is quite mobile, in striking contrast to the other conserved loops that converge at the active site cleft. The catalytic loop (residues 166-171) and the Mg2+ positioning loop (residues 184-186) are a stable part of the large lobe and have low B-factors in all structures solved to date. The stability of the glycine-rich loop is highly dependent on the ligands that occupy the active site cleft with maximum stability achieved in the ternary complex containing Mg x ATP and the peptide inhibitor. In this ternary complex the gamma-phosphate is secured between both lobes by hydrogen bonds to the backbone amide of Ser 53 in the glycine-rich loop and the amino group of Lys 168 in the catalytic loop. In the adenosine ternary complex the water molecule replacing the gamma-phosphate hydrogen bonds between Lys 168 and Asp 166 and makes no contact with the small lobe. This glycine-rich loop is thus the most mobile component of the active site cleft, with the tip of the loop being highly sensitive to what occupies the gamma-subsite.

  20. Sorting cells of the microalga Chlorococcum littorale with increased triacylglycerol productivity.

    PubMed

    Cabanelas, Iago Teles Dominguez; van der Zwart, Mathijs; Kleinegris, Dorinde M M; Wijffels, René H; Barbosa, Maria J

    2016-01-01

    Despite extensive research in the last decades, microalgae are still only economically feasible for high valued markets. Strain improvement is a strategy to increase productivities, hence reducing costs. In this work, we focus on microalgae selection: taking advantage of the natural biological variability of species to select variations based on desired characteristics. We focused on triacylglycerol (TAG), which have applications ranging from biodiesel to high-value omega-3 fatty-acids. Hence, we demonstrated a strategy to sort microalgae cells with increased TAG productivity. 1. We successfully identified sub-populations of cells with increased TAG productivity using Fluorescence assisted cell sorting (FACS). 2. We sequentially sorted cells after repeated cycles of N-starvation, resulting in five sorted populations (S1-S5). 3. The comparison between sorted and original populations showed that S5 had the highest TAG productivity [0.34 against 0.18 g l(-1) day(-1) (original), continuous light]. 4. Original and S5 were compared in lab-scale reactors under simulated summer conditions confirming the increased TAG productivity of S5 (0.4 against 0.2 g l(-1) day(-1)). Biomass composition analyses showed that S5 produced more biomass under N-starvation because of an increase only in TAG content and, flow cytometry showed that our selection removed cells with lower efficiency in producing TAGs. All combined, our results present a successful strategy to improve the TAG productivity of Chlorococcum littorale, without resourcing to genetic manipulation or random mutagenesis. Additionally, the improved TAG productivity of S5 was confirmed under simulated summer conditions, highlighting the industrial potential of S5 for microalgal TAG production.

  1. 2-SR-based electrically small antenna for RFID applications

    NASA Astrophysics Data System (ADS)

    Paredes, Ferran; Zuffanelli, Simone; Aguilà, Pau; Zamora, Gerard; Martin, Ferran; Bonache, Jordi

    2016-04-01

    In this work, the 2-turn spiral resonator (2-SR) is proposed as an electrically small antenna for passive radio frequency identification (RFID) tags at the European ultra-high frequency (UHF) band. The radiation properties are studied in order to explore the viability of the 2-SR applied to tag antenna design. Based on analytical calculations, the radiation pattern is found to provide a cancelation of the radiation nulls. This results in a mitigation of the blind spots in the read range, which are present in typical UHF-RFID tags as an undesired feature. As a proof of concept, a passive tag of size 35 mm × 40 mm (λ 0/10 × λ 0/9) based on the 2-SR antenna is designed and fabricated. Good radiation efficiency (75 %) and a quasi-isotropic radiation pattern are obtained. The experimental tag read range for different directions is in good agreement with the simulation results. The measured read range exhibits maximum and minimum values of 6.7 and 3.5 m, respectively.

  2. Immobilization of proteins onto microbeads using a DNA binding tag for enzymatic assays.

    PubMed

    Kojima, Takaaki; Mizoguchi, Takuro; Ota, Eri; Hata, Jumpei; Homma, Keisuke; Zhu, Bo; Hitomi, Kiyotaka; Nakano, Hideo

    2016-02-01

    A novel DNA-binding protein tag, scCro-tag, which is a single-chain derivative of the bacteriophage lambda Cro repressor, has been developed to immobilize proteins of interest (POI) on a solid support through binding OR consensus DNA (ORC) that is tightly bound by the scCro protein. The scCro-tag successfully bound a transglutaminase 2 (TGase 2) substrate and manganese peroxidase (MnP) to microbeads via scaffolding DNA. The resulting protein-coated microbeads can be utilized for functional analysis of the enzymatic activity using flow cytometry. The quantity of bead-bound proteins can be enhanced by increasing the number of ORCs. In addition, proteins with the scCro-tag that were synthesized using a cell-free protein synthesis system were also immobilized onto the beads, thus indicating that this bead-based system would be applicable to high-throughput analysis of various enzymatic activities. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  3. Quaternary ammonium isobaric tag for a relative and absolute quantification of peptides.

    PubMed

    Setner, Bartosz; Stefanowicz, Piotr; Szewczuk, Zbigniew

    2018-02-01

    Isobaric labeling quantification of peptides has become a method of choice for mass spectrometry-based proteomics studies. However, despite of wide variety of commercially available isobaric tags, none of the currently available methods offers significant improvement of sensitivity of detection during MS experiment. Recently, many strategies were applied to increase the ionization efficiency of peptides involving chemical modifications introducing quaternary ammonium fixed charge. Here, we present a novel quaternary ammonium-based isobaric tag for relative and absolute quantification of peptides (QAS-iTRAQ 2-plex). Upon collisional activation, the new stable benzylic-type cationic reporter ion is liberated from the tag. Deuterium atoms were used to offset the differential masses of a reporter group. We tested the applicability of QAS-iTRAQ 2-plex reagent on a series of model peptides as well as bovine serum albumin tryptic digest. Obtained results suggest usefulness of this isobaric ionization tag for relative and absolute quantification of peptides. Copyright © 2017 John Wiley & Sons, Ltd.

  4. Miniaturization design and implementation of magnetic field coupled RFID antenna

    NASA Astrophysics Data System (ADS)

    Hu, Tiling

    2013-03-01

    The development of internet of things has brought new opportunities and challenges to the application of RFID tags. Moreover, the Miniaturization application trend of tags at present has become the mainstream of development. In this paper, the double-layer design is to reduce the size of HF antenna, and the magnetic null point of magnetic reconnection region between the RLC resonant circuit and the reader provides sufficient energy to the miniaturization of antenna. The calculated and experimental results show that the miniaturization of HF antennas can meet the reading and writing requirement of the international standard ISO/IEC14443 standard. The results of this paper may make a positive contribution to the applications of RFID technology.

  5. 78 FR 32374 - Magnuson-Stevens Act Provisions; General Provisions for Domestic Fisheries; Application for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-30

    ...) 281-9135. FOR FURTHER INFORMATION CONTACT: Jason Berthiaume, Fishery Management Specialist, 978-281...-standard cages and 32-bu (1,703.68 L) cage tags. The traditional cages do not fit on smaller day-boats and... system, with each tag representing 32 bu (1,703.68 L) of allocation, or one cage. In addition to managing...

  6. Application of Cydia pomonella expressed sequence tags: identification and expression of three general odorant binding proteins in codling moth

    USDA-ARS?s Scientific Manuscript database

    The codling moth, Cydia pomonella, is one of the most important pests of pome fruits in the world, yet the molecular genetics and physiology of this insect remains poorly understood. A combined assembly of 8340 expressed sequence tags (ESTs) was generated from Roche 454 GS-FLX sequencing of 8 tissu...

  7. Using XML to Separate Content from the Presentation Software in eLearning Applications

    ERIC Educational Resources Information Center

    Merrill, Paul F.

    2005-01-01

    This paper has shown how XML (extensible Markup Language) can be used to mark up content. Since XML documents, with meaningful tags, can be interpreted easily by humans as well as computers, they are ideal for the interchange of information. Because XML tags can be defined by an individual or organization, XML documents have proven useful in a…

  8. The self-assembly of redox active peptides: Synthesis and electrochemical capacitive behavior.

    PubMed

    Piccoli, Julia P; Santos, Adriano; Santos-Filho, Norival A; Lorenzón, Esteban N; Cilli, Eduardo M; Bueno, Paulo R

    2016-05-01

    The present work reports on the synthesis of a redox-tagged peptide with self-assembling capability aiming applications in electrochemically active capacitive surfaces (associated with the presence of the redox centers) generally useful in electroanalytical applications. Peptide containing ferrocene (fc) molecular (redox) group (Ac-Cys-Ile-Ile-Lys(fc)-Ile-Ile-COOH) was thus synthesized by solid phase peptide synthesis (SPPS). To obtain the electrochemically active capacitive interface, the side chain of the cysteine was covalently bound to the gold electrode (sulfur group) and the side chain of Lys was used to attach the ferrocene in the peptide chain. After obtaining the purified redox-tagged peptide, the self-assembly and redox capability was characterized by cyclic voltammetry (CV) and electrochemical impedance-based capacitance spectroscopy techniques. The obtained results confirmed that the redox-tagged peptide was successfully attached by forming an electroactive self-assembled monolayer onto gold electrode. The design of redox active self-assembly ferrocene-tagged peptide is predictably useful in the development of biosensor devices precisely to detect, in a label-free platform, those biomarkers of clinical relevance. © 2016 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 106: 357-367, 2016. © 2016 Wiley Periodicals, Inc.

  9. Gapped Spectral Dictionaries and Their Applications for Database Searches of Tandem Mass Spectra*

    PubMed Central

    Jeong, Kyowon; Kim, Sangtae; Bandeira, Nuno; Pevzner, Pavel A.

    2011-01-01

    Generating all plausible de novo interpretations of a peptide tandem mass (MS/MS) spectrum (Spectral Dictionary) and quickly matching them against the database represent a recently emerged alternative approach to peptide identification. However, the sizes of the Spectral Dictionaries quickly grow with the peptide length making their generation impractical for long peptides. We introduce Gapped Spectral Dictionaries (all plausible de novo interpretations with gaps) that can be easily generated for any peptide length thus addressing the limitation of the Spectral Dictionary approach. We show that Gapped Spectral Dictionaries are small thus opening a possibility of using them to speed-up MS/MS searches. Our MS-GappedDictionary algorithm (based on Gapped Spectral Dictionaries) enables proteogenomics applications (such as searches in the six-frame translation of the human genome) that are prohibitively time consuming with existing approaches. MS-GappedDictionary generates gapped peptides that occupy a niche between accurate but short peptide sequence tags and long but inaccurate full length peptide reconstructions. We show that, contrary to conventional wisdom, some high-quality spectra do not have good peptide sequence tags and introduce gapped tags that have advantages over the conventional peptide sequence tags in MS/MS database searches. PMID:21444829

  10. Probing the potential of CnaB-type domains for the design of tag/catcher systems

    PubMed Central

    Pröschel, Marlene; Kraner, Max E.; Horn, Anselm H. C.; Schäfer, Lena; Sonnewald, Uwe

    2017-01-01

    Building proteins into larger, post-translational assemblies in a defined and stable way is still a challenging task. A promising approach relies on so-called tag/catcher systems that are fused to the proteins of interest and allow a durable linkage via covalent intermolecular bonds. Tags and catchers are generated by splitting protein domains that contain intramolecular isopeptide or ester bonds that form autocatalytically under physiological conditions. There are already numerous biotechnological and medical applications that demonstrate the usefulness of covalent linkages mediated by these systems. Additional covalent tag/catcher systems would allow creating more complex and ultra-stable protein architectures and networks. Two of the presently available tag/catcher systems were derived from closely related CnaB-domains of Streptococcus pyogenes and Streptococcus dysgalactiae proteins. However, it is unclear whether domain splitting is generally tolerated within the CnaB-family or only by a small subset of these domains. To address this point, we have selected a set of four CnaB domains of low sequence similarity and characterized the resulting tag/catcher systems by computational and experimental methods. Experimental testing for intermolecular isopeptide bond formation demonstrated two of the four systems to be functional. For these two systems length and sequence variations of the peptide tags were investigated revealing only a relatively small effect on the efficiency of the reaction. Our study suggests that splitting into tag and catcher moieties is tolerated by a significant portion of the naturally occurring CnaB-domains, thus providing a large reservoir for the design of novel tag/catcher systems. PMID:28654665

  11. Total Absorption Study of Beta Decays Relevant for Nuclear Applications and Nuclear Structure

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Algora, A.; Valencia, E.; Tain, J. L.

    2014-06-01

    We present an overview of our activities related to the study of the beta decay of neutron rich nuclei relevant for nuclear applications. Recent results of the study of the beta decay of Br using a new segmented total absorption spectrometer are presented. Our measurements were performed at the IGISOL facility using trap-assisted total absorption spectroscopy.

  12. Elastin-like polypeptides: A strategic fusion partner for biologics.

    PubMed

    Yeboah, Agnes; Cohen, Rick I; Rabolli, Charles; Yarmush, Martin L; Berthiaume, Francois

    2016-08-01

    Elastin-like peptides (ELPs) are derivatives of tropoelastin with a unique property that allows them to stay soluble below a certain critical temperature but reversibly form aggregates above that temperature. Since they are derived from tropoelastin, ELPs are biocompatible, non-toxic, and non-immunogenic. The unique properties of ELPs have made them a desirable class of fusion tags used in several biomedical applications including targeted drug delivery and enhancing the half-life of protein drugs. The most significant property of an ELP is that when fused to other proteins, the phase transition property of the ELP is maintained, and the target protein can be purified using the thermally driven property of the ELP. The ELP tag purification process is simple and inexpensive, and involves cycling the protein above and below the transition temperature of the ELP fusion followed by centrifugation to obtain the desired protein, without any need for chromatography. Consequently, using ELPs as a purification tag should be potentially interesting to biopharmaceutical companies who spend a significant percentage of their manufacturing costs on expensive protein purification techniques such as chromatography and filtration. However, ELP tags have not yet been used for commercial protein purification due to some challenges of translating this technique, which has been demonstrated mostly in academic laboratories, to a biotechnology manufacturing environment. The article first reviews the state-of-the-art in protein "ELPylation," and discusses some applications which have benefitted from using ELP as a fusion tag. Then, the article discusses the main advantages of using ELP as a purification tag, and evaluates the remaining hurdles for its implementation in industrial protein production. Biotechnol. Bioeng. 2016;113: 1617-1627. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  13. Robust inference under the beta regression model with application to health care studies.

    PubMed

    Ghosh, Abhik

    2017-01-01

    Data on rates, percentages, or proportions arise frequently in many different applied disciplines like medical biology, health care, psychology, and several others. In this paper, we develop a robust inference procedure for the beta regression model, which is used to describe such response variables taking values in (0, 1) through some related explanatory variables. In relation to the beta regression model, the issue of robustness has been largely ignored in the literature so far. The existing maximum likelihood-based inference has serious lack of robustness against outliers in data and generate drastically different (erroneous) inference in the presence of data contamination. Here, we develop the robust minimum density power divergence estimator and a class of robust Wald-type tests for the beta regression model along with several applications. We derive their asymptotic properties and describe their robustness theoretically through the influence function analyses. Finite sample performances of the proposed estimators and tests are examined through suitable simulation studies and real data applications in the context of health care and psychology. Although we primarily focus on the beta regression models with a fixed dispersion parameter, some indications are also provided for extension to the variable dispersion beta regression models with an application.

  14. Transcriptome-Wide Identification of Preferentially Expressed Genes in the Hypothalamus and Pituitary Gland

    PubMed Central

    St-Amand, Jonny; Yoshioka, Mayumi; Tanaka, Keitaro; Nishida, Yuichiro

    2012-01-01

    To identify preferentially expressed genes in the central endocrine organs of the hypothalamus and pituitary gland, we generated transcriptome-wide mRNA profiles of the hypothalamus, pituitary gland, and parietal cortex in male mice (12–15 weeks old) using serial analysis of gene expression (SAGE). Total counts of SAGE tags for the hypothalamus, pituitary gland, and parietal cortex were 165824, 126688, and 161045 tags, respectively. This represented 59244, 45151, and 55131 distinct tags, respectively. Comparison of these mRNA profiles revealed that 22 mRNA species, including three potential novel transcripts, were preferentially expressed in the hypothalamus. In addition to well-known hypothalamic transcripts, such as hypocretin, several genes involved in hormone function, intracellular transduction, metabolism, protein transport, steroidogenesis, extracellular matrix, and brain disease were identified as preferentially expressed hypothalamic transcripts. In the pituitary gland, 106 mRNA species, including 60 potential novel transcripts, were preferentially expressed. In addition to well-known pituitary genes, such as growth hormone and thyroid stimulating hormone beta, a number of genes classified to function in transport, amino acid metabolism, intracellular transduction, cell adhesion, disulfide bond formation, stress response, transcription, protein synthesis, and turnover, cell differentiation, the cell cycle, and in the cytoskeleton and extracellular matrix were also preferentially expressed. In conclusion, the current study identified not only well-known hypothalamic and pituitary transcripts but also a number of new candidates likely to be involved in endocrine homeostatic systems regulated by the hypothalamus and pituitary gland. PMID:22649398

  15. Peroxisome proliferator-activated receptor-alpha regulates fatty acid utilization in primary human skeletal muscle cells.

    PubMed

    Muoio, Deborah M; Way, James M; Tanner, Charles J; Winegar, Deborah A; Kliewer, Steven A; Houmard, Joseph A; Kraus, William E; Dohm, G Lynis

    2002-04-01

    In humans, skeletal muscle is a major site of peroxisome proliferator-activated receptor-alpha (PPAR-alpha) expression, but its function in this tissue is unclear. We investigated the role of hPPAR-alpha in regulating muscle lipid utilization by studying the effects of a highly selective PPAR-alpha agonist, GW7647, on [(14)C]oleate metabolism and gene expression in primary human skeletal muscle cells. Robust induction of PPAR-alpha protein expression occurred during muscle cell differentiation and corresponded with differentiation-dependent increases in oleate oxidation. In mature myotubes, 48-h treatment with 10-1,000 nmol/l GW7647 increased oleate oxidation dose-dependently, up to threefold. Additionally, GW7647 decreased oleate esterification into myotube triacylglycerol (TAG), up to 45%. This effect was not abolished by etomoxir, a potent inhibitor of beta-oxidation, indicating that PPAR-alpha-mediated TAG depletion does not depend on reciprocal changes in fatty acid catabolism. Consistent with its metabolic actions, GW7647 induced mRNA expression of mitochondrial enzymes that promote fatty acid catabolism; carnitine palmityltransferase 1 and malonyl-CoA decarboxylase increased approximately 2-fold, whereas pyruvate dehydrogenase kinase 4 increased 45-fold. Expression of several genes that regulate glycerolipid synthesis was not changed by GW7647 treatment, implicating involvement of other targets to explain the TAG-depleting effect of the compound. These results demonstrate a role for hPPAR-alpha in regulating muscle lipid homeostasis.

  16. Transcriptome-wide identification of preferentially expressed genes in the hypothalamus and pituitary gland.

    PubMed

    St-Amand, Jonny; Yoshioka, Mayumi; Tanaka, Keitaro; Nishida, Yuichiro

    2011-01-01

    To identify preferentially expressed genes in the central endocrine organs of the hypothalamus and pituitary gland, we generated transcriptome-wide mRNA profiles of the hypothalamus, pituitary gland, and parietal cortex in male mice (12-15 weeks old) using serial analysis of gene expression (SAGE). Total counts of SAGE tags for the hypothalamus, pituitary gland, and parietal cortex were 165824, 126688, and 161045 tags, respectively. This represented 59244, 45151, and 55131 distinct tags, respectively. Comparison of these mRNA profiles revealed that 22 mRNA species, including three potential novel transcripts, were preferentially expressed in the hypothalamus. In addition to well-known hypothalamic transcripts, such as hypocretin, several genes involved in hormone function, intracellular transduction, metabolism, protein transport, steroidogenesis, extracellular matrix, and brain disease were identified as preferentially expressed hypothalamic transcripts. In the pituitary gland, 106 mRNA species, including 60 potential novel transcripts, were preferentially expressed. In addition to well-known pituitary genes, such as growth hormone and thyroid stimulating hormone beta, a number of genes classified to function in transport, amino acid metabolism, intracellular transduction, cell adhesion, disulfide bond formation, stress response, transcription, protein synthesis, and turnover, cell differentiation, the cell cycle, and in the cytoskeleton and extracellular matrix were also preferentially expressed. In conclusion, the current study identified not only well-known hypothalamic and pituitary transcripts but also a number of new candidates likely to be involved in endocrine homeostatic systems regulated by the hypothalamus and pituitary gland.

  17. Kinetic Studies of Inhibition of the Aβ(1–42) Aggregation Using a Ferrocene-tagged β-Sheet Breaker Peptide

    PubMed Central

    Zhang, Lin; Yagnik, Gargey; Peng, Yong; Wang, Jianxiu; Xu, H. Howard; Hao, Yuanqiang; Liu, You-Nian; Zhou, Feimeng

    2013-01-01

    The aggregation of amyloidogenic proteins/peptides has been closely linked to the neuropathology of several important neurological disorders. In Alzheimer's disease (AD), amyloid beta (Aβ) peptides and their aggregation are believed to be at least partially responsible for the etiology of AD. The aggregate-inflicted cellular toxicity can be inhibited by short peptides whose sequence are homologous to segments of the Aβ(1–42) peptide responsible for β-sheet stacking (referred to as the β-sheet breaker peptides). Herein a water-soluble ferrocene (Fc)-tagged β-sheet breaker peptide (Fc-KLVFFK6) is used as an electrochemical probe for kinetic studies of the inhibition of the Aβ(1–42) fibrillation process and for determination of the optimal concentration of β-sheet breaker peptide for efficient inhibition. Our results demonstrated that Fc-KLVFFK6 interacts with the Aβ aggregates instantaneously in solution, and sub-stoichiometric amount of Fc-KLVFFK6 is sufficient to inhibit the formation of the Aβ oligomers and fibrils and to reduce the toxicity of Aβ(1–42). The interaction between Fc-KLVFFK6 and Aβ(1–42) follows a pseudo-first-order reaction, with a rate constant of 1.89 ± 0.05 × 10−4 s−1. Tagging β-sheet breaker peptides with a redox label facilitates design, screening, and rational use of peptidic inhibitors for impeding/altering Aβ aggregation. PMID:23232068

  18. The role of IL-6 and IL-1beta in painful perineural inflammatory neuritis.

    PubMed

    Eliav, Eli; Benoliel, Rafael; Herzberg, Uri; Kalladka, Mythili; Tal, Michael

    2009-05-01

    Inflammation along a nerve trunk (perineural inflammation), without detectable axonal damage, has been shown to induce transient pain in the organ supplied by the nerve. The aims of the present study were to study the role IL-6 and IL-1beta, in pain induced by perineural inflammation. IL-6 and IL-1beta secretion from rat's sciatic nerves, L-5 Dorsal Root Ganglia (DRG), and the hind paw skin, 3 and 8 days following exposure of the nerve to Complete Freund's Adjuvant (CFA), were measured using ELISA method. Hind paw tactile-allodynia, mechano-hyperalgesia, heat-allodynia and electrical detection thresholds were tested up to 8 days following the application of CFA, IL-6 or IL-1beta adjacent to the sciatic nerve trunk. Employing electrophysiological recording, saphenous nerve spontaneous activity, nerve trunk mechano-sensitivity and paw tactile detection threshold (determined by recording action potential induced by the lowest mechanical stimulus) were assessed 3 and 8 days following exposure of the nerve trunk to CFA, IL-6, or IL-1beta. IL-6 and IL-1beta secretion from the nerve was significantly elevated on the 3rd day post-operation (DPO). On the 8th DPO, IL-6 levels returned to baseline while IL-1beta levels remained significantly elevated. The DRG cytokine's level was increased on the 3rd and 8th DPOs, contralateral cytokine's level was increased on the 3rd DPO. The skin IL-6 level was increased bilaterally on the 3rd DPO and returned to baseline on the 8th DPO. IL-1beta levels increased in the affected side on the 3rd and bilaterally on the 8th DPO. Direct application of IL-6 or CFA on the sciatic nerve induced significant hind paw tactile-allodynia from the 1st to 5th DPOs, reduced electrical detection threshold from the 1st to 3rd DPOs, mechano-hyperalgesia from 3rd to 5th DPOs and heat-allodynia on the 3rd DPO. Direct application of IL-1beta induced paw tactile and heat-allodynia on the 7-8th DPOs and mechano-hyperalgesia on the 5-8th DPOs. Perineural inflammation significantly increased spontaneous activity myelinated fibres 3 and 8 days following the application. Direct application of IL-6 induced elevation of spontaneous activity on the 3rd while IL-1beta on the 8th DPO. Nerve mechano-sensitivity was significantly increased on the 3rd day following exposure to CFA and IL-6 and on the 8th following CFA application. The rat's paw lowest mechanical force necessary for induction of action potential, was significantly reduced 3 days following CFA application. IL-6 and IL-1beta play an important role in pain induced by perineural inflammation. IL-6 activity is more prominent immediately following application (2-5th DPOs), while IL-1beta, activity is more significant in a later stage (5-8th DPOs).

  19. Wireless Hydrogen Smart Sensor Based on Pt/Graphene-Immobilized Radio-Frequency Identification Tag.

    PubMed

    Lee, Jun Seop; Oh, Jungkyun; Jun, Jaemoon; Jang, Jyongsik

    2015-08-25

    Hydrogen, a clean-burning fuel, is of key importance to various industrial applications, including fuel cells and the aerospace and automotive industries. However, hydrogen gas is odorless, colorless, and highly flammable; thus, appropriate safety protocol implementation and monitoring are essential. Highly sensitive hydrogen-gas leak detection and surveillance systems are needed; additionally, the ability to monitor large areas (e.g., cities) via wireless networks is becoming increasingly important. In this report, we introduce a radio frequency identification (RFID)-based wireless smart-sensor system, composed of a Pt-decorated reduced graphene oxide (Pt_rGO)-immobilized RFID sensor tag and an RFID-reader antenna-connected network analyzer to detect hydrogen gas. The Pt_rGOs, produced using a simple chemical reduction process, were immobilized on an antenna pattern in the sensor tag through spin coating. The resulting Pt_rGO-based RFID sensor tag exhibited a high sensitivity to hydrogen gas at unprecedentedly low concentrations (1 ppm), with wireless communication between the sensor tag and RFID-reader antenna. The wireless sensor tag demonstrated flexibility and a long lifetime due to the strong immobilization of Pt_rGOs on the substrate and battery-independent operation during hydrogen sensing, respectively.

  20. Single molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes

    PubMed Central

    Barlag, Britta; Beutel, Oliver; Janning, Dennis; Czarniak, Frederik; Richter, Christian P.; Kommnick, Carina; Göser, Vera; Kurre, Rainer; Fabiani, Florian; Erhardt, Marc; Piehler, Jacob; Hensel, Michael

    2016-01-01

    The investigation of the subcellular localization, dynamics and interaction of proteins and protein complexes in prokaryotes is complicated by the small size of the cells. Super-resolution microscopy (SRM) comprise various new techniques that allow light microscopy with a resolution that can be up to ten-fold higher than conventional light microscopy. Application of SRM techniques to living prokaryotes demands the introduction of suitable fluorescent probes, usually by fusion of proteins of interest to fluorescent proteins with properties compatible to SRM. Here we describe an approach that is based on the genetically encoded self-labelling enzymes HaloTag and SNAP-tag. Proteins of interest are fused to HaloTag or SNAP-tag and cell permeable substrates can be labelled with various SRM-compatible fluorochromes. Fusions of the enzyme tags to subunits of a type I secretion system (T1SS), a T3SS, the flagellar rotor and a transcription factor were generated and analysed in living Salmonella enterica. The new approach is versatile in tagging proteins of interest in bacterial cells and allows to determine the number, relative subcellular localization and dynamics of protein complexes in living cells. PMID:27534893

  1. Sequence tagging reveals unexpected modifications in toxicoproteomics

    PubMed Central

    Dasari, Surendra; Chambers, Matthew C.; Codreanu, Simona G.; Liebler, Daniel C.; Collins, Ben C.; Pennington, Stephen R.; Gallagher, William M.; Tabb, David L.

    2010-01-01

    Toxicoproteomic samples are rich in posttranslational modifications (PTMs) of proteins. Identifying these modifications via standard database searching can incur significant performance penalties. Here we describe the latest developments in TagRecon, an algorithm that leverages inferred sequence tags to identify modified peptides in toxicoproteomic data sets. TagRecon identifies known modifications more effectively than the MyriMatch database search engine. TagRecon outperformed state of the art software in recognizing unanticipated modifications from LTQ, Orbitrap, and QTOF data sets. We developed user-friendly software for detecting persistent mass shifts from samples. We follow a three-step strategy for detecting unanticipated PTMs in samples. First, we identify the proteins present in the sample with a standard database search. Next, identified proteins are interrogated for unexpected PTMs with a sequence tag-based search. Finally, additional evidence is gathered for the detected mass shifts with a refinement search. Application of this technology on toxicoproteomic data sets revealed unintended cross-reactions between proteins and sample processing reagents. Twenty five proteins in rat liver showed signs of oxidative stress when exposed to potentially toxic drugs. These results demonstrate the value of mining toxicoproteomic data sets for modifications. PMID:21214251

  2. A Secure RFID Tag Authentication Protocol with Privacy Preserving in Telecare Medicine Information System.

    PubMed

    Li, Chun-Ta; Weng, Chi-Yao; Lee, Cheng-Chi

    2015-08-01

    Radio Frequency Identification (RFID) based solutions are widely used for providing many healthcare applications include patient monitoring, object traceability, drug administration system and telecare medicine information system (TMIS) etc. In order to reduce malpractices and ensure patient privacy, in 2015, Srivastava et al. proposed a hash based RFID tag authentication protocol in TMIS. Their protocol uses lightweight hash operation and synchronized secret value shared between back-end server and tag, which is more secure and efficient than other related RFID authentication protocols. Unfortunately, in this paper, we demonstrate that Srivastava et al.'s tag authentication protocol has a serious security problem in that an adversary may use the stolen/lost reader to connect to the medical back-end server that store information associated with tagged objects and this privacy damage causing the adversary could reveal medical data obtained from stolen/lost readers in a malicious way. Therefore, we propose a secure and efficient RFID tag authentication protocol to overcome security flaws and improve the system efficiency. Compared with Srivastava et al.'s protocol, the proposed protocol not only inherits the advantages of Srivastava et al.'s authentication protocol for TMIS but also provides better security with high system efficiency.

  3. Application of selected ion monitoring to the analysis of triacylglycerols in olive oil by high temperature-gas chromatography/mass spectrometry.

    PubMed

    Ruiz-Samblás, C; González-Casado, A; Cuadros-Rodríguez, L; García, F P Rodríguez

    2010-06-30

    The analysis of the triacylglycerol (TAG) composition of oils is a very challenging task, since the TAGs have very similar physico-chemical properties. In this work, a high temperature-gas chromatographic method coupled to electron ionization-mass spectrometry (HT-GC/EI-MS), in the Selected Ion Monitoring (SIM) mode, method was developed for the analysis of TAGs in the olive oil; this is a method suitable for routine analysis. This method was developed using commercially available standard TAGs. The TAGs studied were separated according to their equivalent carbon number and degree of unsaturation. The peak assignment was carried out by locating the characteristic fragment ions having the same retention time on the SIM profile such as [RCO+74](+) and [RCO+128](+) ions, due to the fatty acyl residues on sn-1, sn-2 and sn-3 positions of the TAG molecule and the [M-OCOR](+) ions corresponding to the acyl ions. The developed method was very useful to eliminate the interferences that appeared in the mass spectrum since electron ionization can prevent satisfactory interpretation of spectra. Copyright 2010 Elsevier B.V. All rights reserved.

  4. Application of coupled affinity-sizing chromatography for the detection of proteolyzed HSA-tagged proteins.

    PubMed

    London, Anne Serdakowski; Patel, Kunal; Quinn, Lisa; Lemmerer, Martin

    2015-04-01

    Coupled affinity liquid chromatography and size exclusion chromatography (ALC-SEC) is a technique that has been shown to successfully report product quality of proteins during cell expression and prior to the commencement of downstream processing chromatography steps. This method was applied to monitoring the degradation and subsequent partial remediation of a HSA-tagged protein which showed proteolysis, allowing for rapid cell line development to address this product quality dilemma. This paper outlines the novel application of this method for measuring and addressing protease-induced proteolysis. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. High data density and capacity in chipless radiofrequency identification (chipless-RFID) tags based on double-chains of S-shaped split ring resonators (S-SRRs)

    NASA Astrophysics Data System (ADS)

    Herrojo, Cristian; Mata-Contreras, Javier; Paredes, Ferran; Martín, Ferran

    2017-11-01

    The data density per surface (DPS) is a figure of merit in chipless radiofrequency identification (chipless-RFID) tags. In this paper, it is demonstrated that chipless-RFID tags with high DPS can be implemented by using double-chains of S-shaped split ring resonators (S-SRRs). Tag reading is achieved by near-field coupling between the tag and the reader, a CPW transmission line fed by a harmonic signal tuned to the resonance frequency of the S-SRRs. By transversally displacing the tag over the CPW, the transmission coefficient of the line is modulated by tag motion. This effectively modulates the amplitude of the injected (carrier) signal at the output port of the line, and the identification (ID) code, determined by the presence or absence of S-SRRs at predefined and equidistant positions in the chains, is contained in the envelope function. The DPS is determined by S-SRR dimensions and by the distance between S-SRRs in the chains. However, by using two chains of S-SRRs, the number of bits per unit length that can be accommodated is very high. This chipless-RFID system is of special interest in applications where the reading distance can be sacrificed in favor of data capacity (e.g., security and authentication). Encoding of corporate documents, ballots, exams, etc., by directly printing the proposed tags on the item product to prevent counterfeiting is envisaged.

  6. Immobilized palladium(II) ion affinity chromatography for recovery of recombinant proteins with peptide tags containing histidine and cysteine.

    PubMed

    Kikot, Pamela; Polat, Aise; Achilli, Estefania; Fernandez Lahore, Marcelo; Grasselli, Mariano

    2014-11-01

    Fusion of peptide-based tags to recombinant proteins is currently one of the most used tools for protein production. Also, immobilized metal ion affinity chromatography (IMAC) has a huge application in protein purification, especially in research labs. The combination of expression systems of recombinant tagged proteins with this robust chromatographic system has become an efficient and rapid tool to produce milligram-range amounts of proteins. IMAC-Ni(II) columns have become the natural partners of 6xHis-tagged proteins. The Ni(II) ion is considered as the best compromise of selectivity and affinity for purification of a recombinant His-tagged protein. The palladium(II) ion is also able to bind to side chains of amino acids and form ternary complexes with iminodiacetic acid and free amino acids and other sulfur-containing molecules. In this work, we evaluated two different cysteine- and histidine-containing six amino acid tags linked to the N-terminal group of green fluorescent protein (GFP) and studied the adsorption and elution conditions using novel eluents. Both cysteine-containing tagged GFPs were able to bind to IMAC-Pd(II) matrices and eluted successfully using a low concentration of thiourea solution. The IMAC-Ni(II) system reaches less than 20% recovery of the cysteine-containing tagged GFP from a crude homogenate of recombinant Escherichia coli, meanwhile the IMAC-Pd(II) yields a recovery of 45% with a purification factor of 13. Copyright © 2014 John Wiley & Sons, Ltd.

  7. Optimal Detection Range of RFID Tag for RFID-based Positioning System Using the k-NN Algorithm.

    PubMed

    Han, Soohee; Kim, Junghwan; Park, Choung-Hwan; Yoon, Hee-Cheon; Heo, Joon

    2009-01-01

    Positioning technology to track a moving object is an important and essential component of ubiquitous computing environments and applications. An RFID-based positioning system using the k-nearest neighbor (k-NN) algorithm can determine the position of a moving reader from observed reference data. In this study, the optimal detection range of an RFID-based positioning system was determined on the principle that tag spacing can be derived from the detection range. It was assumed that reference tags without signal strength information are regularly distributed in 1-, 2- and 3-dimensional spaces. The optimal detection range was determined, through analytical and numerical approaches, to be 125% of the tag-spacing distance in 1-dimensional space. Through numerical approaches, the range was 134% in 2-dimensional space, 143% in 3-dimensional space.

  8. Tag Content Access Control with Identity-based Key Exchange

    NASA Astrophysics Data System (ADS)

    Yan, Liang; Rong, Chunming

    2010-09-01

    Radio Frequency Identification (RFID) technology that used to identify objects and users has been applied to many applications such retail and supply chain recently. How to prevent tag content from unauthorized readout is a core problem of RFID privacy issues. Hash-lock access control protocol can make tag to release its content only to reader who knows the secret key shared between them. However, in order to get this shared secret key required by this protocol, reader needs to communicate with a back end database. In this paper, we propose to use identity-based secret key exchange approach to generate the secret key required for hash-lock access control protocol. With this approach, not only back end database connection is not needed anymore, but also tag cloning problem can be eliminated at the same time.

  9. Application of the accurate mass and time tag approach in studies of the human blood lipidome

    PubMed Central

    Ding, Jie; Sorensen, Christina M.; Jaitly, Navdeep; Jiang, Hongliang; Orton, Daniel J.; Monroe, Matthew E.; Moore, Ronald J.; Smith, Richard D.; Metz, Thomas O.

    2008-01-01

    We report a preliminary demonstration of the accurate mass and time (AMT) tag approach for lipidomics. Initial data-dependent LC-MS/MS analyses of human plasma, erythrocyte, and lymphocyte lipids were performed in order to identify lipid molecular species in conjunction with complementary accurate mass and isotopic distribution information. Identified lipids were used to populate initial lipid AMT tag databases containing 250 and 45 entries for those species detected in positive and negative electrospray ionization (ESI) modes, respectively. The positive ESI database was then utilized to identify human plasma, erythrocyte, and lymphocyte lipids in high-throughput LC-MS analyses based on the AMT tag approach. We were able to define the lipid profiles of human plasma, erythrocytes, and lymphocytes based on qualitative and quantitative differences in lipid abundance. PMID:18502191

  10. Highly efficient purification of protein complexes from mammalian cells using a novel streptavidin-binding peptide and hexahistidine tandem tag system: Application to Bruton's tyrosine kinase

    PubMed Central

    Li, Yifeng; Franklin, Sarah; Zhang, Michael J; Vondriska, Thomas M

    2011-01-01

    Tandem affinity purification (TAP) is a generic approach for the purification of protein complexes. The key advantage of TAP is the engineering of dual affinity tags that, when attached to the protein of interest, allow purification of the target protein along with its binding partners through two consecutive purification steps. The tandem tag used in the original method consists of two IgG-binding units of protein A from Staphylococcus aureus (ProtA) and the calmodulin-binding peptide (CBP), and it allows for recovery of 20–30% of the bait protein in yeast. When applied to higher eukaryotes, however, this classical TAP tag suffers from low yields. To improve protein recovery in systems other than yeast, we describe herein the development of a three-tag system comprised of CBP, streptavidin-binding peptide (SBP) and hexa-histidine. We illustrate the application of this approach for the purification of human Bruton's tyrosine kinase (Btk), which results in highly efficient binding and elution of bait protein in both purification steps (>50% recovery). Combined with mass spectrometry for protein identification, this TAP strategy facilitated the first nonbiased analysis of Btk interacting proteins. The high efficiency of the SBP-His6 purification allows for efficient recovery of protein complexes formed with a target protein of interest from a small amount of starting material, enhancing the ability to detect low abundance and transient interactions in eukaryotic cell systems. PMID:21080425

  11. Novel, continuous monitoring of fine‐scale movement using fixed‐position radiotelemetry arrays and random forest location fingerprinting

    USGS Publications Warehouse

    Harbicht, Andrew B.; Castro-Santos, Theodore R.; Ardren, William R.; Gorsky, Dimitry; Fraser, Dylan

    2017-01-01

    Radio‐tag signals from fixed‐position antennas are most often used to indicate presence or absence of individuals, or to estimate individual activity levels from signal strength variation within an antenna's detection zone. The potential of such systems to provide more precise information on tag location and movement has not been explored in great detail in an ecological setting.By reversing the roles that transmitters and receivers play in localization methods common to the telecommunications industry, we present a new telemetric tool for accurately estimating the location of tagged individuals from received signal strength values. The methods used to characterize the study area in terms of received signal strength are described, as is the random forest model used for localization. The resulting method is then validated using test data before being applied to true data collected from tagged individuals in the study site.Application of the localization method to test data withheld from the learning dataset indicated a low average error over the entire study area (<1 m), whereas application of the localization method to real data produced highly probable results consistent with field observations.This telemetric approach provided detailed movement data for tagged fish along a single axis (a migratory path) and is particularly useful for monitoring passage along migratory routes. The new methods applied in this study can also be expanded to include multiple axes (x, y, z) and multiple environments (aquatic and terrestrial) for remotely monitoring wildlife movement.

  12. Signal and array processing techniques for RFID readers

    NASA Astrophysics Data System (ADS)

    Wang, Jing; Amin, Moeness; Zhang, Yimin

    2006-05-01

    Radio Frequency Identification (RFID) has recently attracted much attention in both the technical and business communities. It has found wide applications in, for example, toll collection, supply-chain management, access control, localization tracking, real-time monitoring, and object identification. Situations may arise where the movement directions of the tagged RFID items through a portal is of interest and must be determined. Doppler estimation may prove complicated or impractical to perform by RFID readers. Several alternative approaches, including the use of an array of sensors with arbitrary geometry, can be applied. In this paper, we consider direction-of-arrival (DOA) estimation techniques for application to near-field narrowband RFID problems. Particularly, we examine the use of a pair of RFID antennas to track moving RFID tagged items through a portal. With two antennas, the near-field DOA estimation problem can be simplified to a far-field problem, yielding a simple way for identifying the direction of the tag movement, where only one parameter, the angle, needs to be considered. In this case, tracking of the moving direction of the tag simply amounts to computing the spatial cross-correlation between the data samples received at the two antennas. It is pointed out that the radiation patterns of the reader and tag antennas, particularly their phase characteristics, have a significant effect on the performance of DOA estimation. Indoor experiments are conducted in the Radar Imaging and RFID Labs at Villanova University for validating the proposed technique for target movement direction estimations.

  13. An Active RFID Accountability System (RAS) for Constrained Wireless Environments

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Barker, Alan M; Hanson, Gregory R; Sexton, Angela Kay

    A team from Oak Ridge National Laboratory (ORNL) has developed an RFID Accountability System (RAS) that allows items with active RFID tags to be tracked in environments where tags may not be able to transmit their location continuously. The system uses activators that transmit a short range signal. Active RFID tags are in a sleep state until they encounter an activator. Then they transmit a signal that is picked up by the antennas installed throughout the building. This paper presents the theory of operation, application areas, lessons learned, and key features developed over the course of seven years of developmentmore » and use.« less

  14. Genome-wide analysis of oleosin gene family in 22 tree species: An accelerator for metabolic engineering of biofuel crops and agrigenomics industrial applications?

    USDA-ARS?s Scientific Manuscript database

    Triacylglycerols (TAGs) are the major molecules of energy storage in eukaryotes. Trees contribute to part of enormous plant oil reserves because fruits and kernels of many trees contain up to 50-80% of oil. TAGs accumulate in oil bodies in plants, similar to oil droplets in animals. Oleosins (OLEs) ...

  15. Detection, Identification, Location, and Remote Sensing Using SAW RFID Sensor Tags

    NASA Technical Reports Server (NTRS)

    Barton, Richard J.; Kennedy, Timothy F.; Williams, Robert M.; Fink, Patrick W.; Ngo, Phong H.

    2009-01-01

    The Electromagnetic Systems Branch (EV4) of the Avionic Systems Division at NASA Johnson Space Center in Houston, TX is studying the utility of surface acoustic wave (SAW) radiofrequency identification (RFID) tags for multiple wireless applications including detection, identification, tracking, and remote sensing of objects on the lunar surface, monitoring of environmental test facilities, structural shape and health monitoring, and nondestructive test and evaluation of assets. For all of these applications, it is anticipated that the system utilized to interrogate the SAW RFID tags may need to operate at fairly long range and in the presence of considerable multipath and multiple-access interference. Towards that end, EV4 is developing a prototype SAW RFID wireless interrogation system for use in such environments called the Passive Adaptive RFID Sensor Equipment (PARSED) system. The system utilizes a digitally beam-formed planar receiving antenna array to extend range and provide direction-of-arrival information coupled with an approximate maximum-likelihood signal processing algorithm to provide near-optimal estimation of both range and temperature. The system is capable of forming a large number of beams within the field of view and resolving the information from several tags within each beam. The combination of both spatial and waveform discrimination provides the capability to track and monitor telemetry from a large number of objects appearing simultaneously within the field of view of the receiving array. In this paper, we will consider the application of the PARSEQ system to the problem of simultaneous detection, identification, localization, and temperature estimation for multiple objects. We will summarize the overall design of the PARSEQ system and present a detailed description of the design and performance of the signal detection and estimation algorithms incorporated in the system. The system is currently configured only to measure temperature (jointly with range and tag ID), but future versions will be revised to measure parameters other than temperature as SAW tags capable of interfacing with external sensors become available. It is anticipated that the estimation of arbitrary parameters measured using SAW-based sensors will be based on techniques very similar to the joint range and temperature estimation techniques described in this paper.

  16. 'Tagger' - a Mac OS X Interactive Graphical Application for Data Inference and Analysis of N-Dimensional Datasets in the Natural Physical Sciences.

    NASA Astrophysics Data System (ADS)

    Morse, P. E.; Reading, A. M.; Lueg, C.

    2014-12-01

    Pattern-recognition in scientific data is not only a computational problem but a human-observer problem as well. Human observation of - and interaction with - data visualization software can augment, select, interrupt and modify computational routines and facilitate processes of pattern and significant feature recognition for subsequent human analysis, machine learning, expert and artificial intelligence systems.'Tagger' is a Mac OS X interactive data visualisation tool that facilitates Human-Computer interaction for the recognition of patterns and significant structures. It is a graphical application developed using the Quartz Composer framework. 'Tagger' follows a Model-View-Controller (MVC) software architecture: the application problem domain (the model) is to facilitate novel ways of abstractly representing data to a human interlocutor, presenting these via different viewer modalities (e.g. chart representations, particle systems, parametric geometry) to the user (View) and enabling interaction with the data (Controller) via a variety of Human Interface Devices (HID). The software enables the user to create an arbitrary array of tags that may be appended to the visualised data, which are then saved into output files as forms of semantic metadata. Three fundamental problems that are not strongly supported by conventional scientific visualisation software are addressed:1] How to visually animate data over time, 2] How to rapidly deploy unconventional parametrically driven data visualisations, 3] How to construct and explore novel interaction models that capture the activity of the end-user as semantic metadata that can be used to computationally enhance subsequent interrogation. Saved tagged data files may be loaded into Tagger, so that tags may be tagged, if desired. Recursion opens up the possibility of refining or overlapping different types of tags, tagging a variety of different POIs or types of events, and of capturing different types of specialist observations of important or noticeable events. Other visualisations and modes of interaction will also be demonstrated, with the aim of discovering knowledge in large datasets in the natural, physical sciences. Fig.1 Wave height data from an oceanographic Wave Rider Buoy. Colors/radii are driven by wave height data.

  17. Multi-Residential Activity Labelling in Smart Homes with Wearable Tags Using BLE Technology

    PubMed Central

    Mokhtari, Ghassem; Zhang, Qing; Karunanithi, Mohanraj

    2018-01-01

    Smart home platforms show promising outcomes to provide a better quality of life for residents in their homes. One of the main challenges that exists with these platforms in multi-residential houses is activity labeling. As most of the activity sensors do not provide any information regarding the identity of the person who triggers them, it is difficult to label the sensor events in multi-residential smart homes. To deal with this challenge, individual localization in different areas can be a promising solution. The localization information can be used to automatically label the activity sensor data to individuals. Bluetooth low energy (BLE) is a promising technology for this application due to how easy it is to implement and its low energy footprint. In this approach, individuals wear a tag that broadcasts its unique identity (ID) in certain time intervals, while fixed scanners listen to the broadcasting packet to localize the tag and the individual. However, the localization accuracy of this method depends greatly on different settings of broadcasting signal strength, and the time interval of BLE tags. To achieve the best localization accuracy, this paper studies the impacts of different advertising time intervals and power levels, and proposes an efficient and applicable algorithm to select optimal value settings of BLE sensors. Moreover, it proposes an automatic activity labeling method, through integrating BLE localization information and ambient sensor data. The applicability and effectiveness of the proposed structure is also demonstrated in a real multi-resident smart home scenario. PMID:29562666

  18. Multi-Residential Activity Labelling in Smart Homes with Wearable Tags Using BLE Technology.

    PubMed

    Mokhtari, Ghassem; Anvari-Moghaddam, Amjad; Zhang, Qing; Karunanithi, Mohanraj

    2018-03-19

    Smart home platforms show promising outcomes to provide a better quality of life for residents in their homes. One of the main challenges that exists with these platforms in multi-residential houses is activity labeling. As most of the activity sensors do not provide any information regarding the identity of the person who triggers them, it is difficult to label the sensor events in multi-residential smart homes. To deal with this challenge, individual localization in different areas can be a promising solution. The localization information can be used to automatically label the activity sensor data to individuals. Bluetooth low energy (BLE) is a promising technology for this application due to how easy it is to implement and its low energy footprint. In this approach, individuals wear a tag that broadcasts its unique identity (ID) in certain time intervals, while fixed scanners listen to the broadcasting packet to localize the tag and the individual. However, the localization accuracy of this method depends greatly on different settings of broadcasting signal strength, and the time interval of BLE tags. To achieve the best localization accuracy, this paper studies the impacts of different advertising time intervals and power levels, and proposes an efficient and applicable algorithm to select optimal value settings of BLE sensors. Moreover, it proposes an automatic activity labeling method, through integrating BLE localization information and ambient sensor data. The applicability and effectiveness of the proposed structure is also demonstrated in a real multi-resident smart home scenario.

  19. Improving Efficiency of Passive RFID Tag Anti-Collision Protocol Using Dynamic Frame Adjustment and Optimal Splitting.

    PubMed

    Memon, Muhammad Qasim; He, Jingsha; Yasir, Mirza Ammar; Memon, Aasma

    2018-04-12

    Radio frequency identification is a wireless communication technology, which enables data gathering and identifies recognition from any tagged object. The number of collisions produced during wireless communication would lead to a variety of problems including unwanted number of iterations and reader-induced idle slots, computational complexity in terms of estimation as well as recognition of the number of tags. In this work, dynamic frame adjustment and optimal splitting are employed together in the proposed algorithm. In the dynamic frame adjustment method, the length of frames is based on the quantity of tags to yield optimal efficiency. The optimal splitting method is conceived with smaller duration of idle slots using an optimal value for splitting level M o p t , where (M > 2), to vary slot sizes to get the minimal identification time for the idle slots. The application of the proposed algorithm offers the advantages of not going for the cumbersome estimation of the quantity of tags incurred and the size (number) of tags has no effect on its performance efficiency. Our experiment results show that using the proposed algorithm, the efficiency curve remains constant as the number of tags varies from 50 to 450, resulting in an overall theoretical gain in the efficiency of 0.032 compared to system efficiency of 0.441 and thus outperforming both dynamic binary tree slotted ALOHA (DBTSA) and binary splitting protocols.

  20. Affinity Purification of Proteins in Tag-Free Form: Split Intein-Mediated Ultrarapid Purification (SIRP).

    PubMed

    Guan, Dongli; Chen, Zhilei

    2017-01-01

    Proteins purified using affinity-based chromatography often exploit a recombinant affinity tag. Existing methods for the removal of the extraneous tag, needed for many applications, suffer from poor efficiency and/or high cost. Here we describe a simple, efficient, and potentially low-cost approach-split intein-mediated ultrarapid purification (SIRP)-for both the purification of the desired tagged protein from Escherichia coli lysate and removal of the tag in less than 1 h. The N- and C-fragment of a self-cleaving variant of a naturally split DnaE intein from Nostoc punctiforme are genetically fused to the N-terminus of an affinity tag and a protein of interest (POI), respectively. The N-intein/affinity tag is used to functionalize an affinity resin. The high affinity between the N- and C-fragment of DnaE intein enables the POI to be purified from the lysate via affinity to the resin, and the intein-mediated C-terminal cleavage reaction causes tagless POI to be released into the flow-through. The intein cleavage reaction is strongly inhibited by divalent ions (e.g., Zn 2+ ) under non-reducing conditions and is significantly enhanced by reducing conditions. The POI is cleaved efficiently regardless of the identity of the N-terminal amino acid except in the cases of threonine and proline, and the N-intein-functionalized affinity resin can be regenerated for multiple cycles of use.

  1. Graphene oxide wrapped SERS tags: multifunctional platforms toward optical labeling, photothermal ablation of bacteria, and the monitoring of killing effect.

    PubMed

    Lin, Donghai; Qin, Tianqi; Wang, Yunqing; Sun, Xiuyan; Chen, Lingxin

    2014-01-22

    As novel optical nanoprobes, surface-enhanced Raman scattering (SERS) tags have drawn growing interests in the application of biomedical imaging and phototherapies. Herein, we demonstrated a novel in situ synthesis strategy for GO wrapped gold nanocluster SERS tags by using a tris(2,2'-bipyridyl)ruthenium(II) chloride (Rubpy)/GO nanohybrid as a complex Raman reporter, inspired by the role of GO as an artificial receptor for various dyes. The introduction of GO in the synthesis procedure provided systematic solutions for controlling several key parameters of SERS tags, including reproducibility, sensitivity, and colloidal and signal stability. An additional interesting thermal-sensitive SERS property (SERS intensity decreased upon increasing the temperature) was also achieved due to the heat-induced release/redistribution of reporter molecules adsorbed on GO. Combining the synergic effect of these features, we further fabricated multifunctional, aldehyde group conjugated Au@Rubpy/GO SERS tags for optical labeling and photothermal ablation of bacteria. Sensitive Raman imaging of gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria could be realized, and satisfactory photothermal killing efficacy for both bacteria was achieved. Our results also demonstrated the correlation among the SERS intensity decrease ratio, bacteria survival rate, and the terminal temperature of the tag-bacteria suspension, showing the possibility to use SERS assay to measure antibacterial response during the photothermal process using this tag.

  2. Enabling Tools and Methods for International, Inter-disciplinary and Educational Collaboration

    NASA Astrophysics Data System (ADS)

    Robinson, E. M.; Hoijarvi, K.; Falke, S.; Fialkowski, E.; Kieffer, M.; Husar, R. B.

    2008-05-01

    In the past, collaboration has taken place in tightly-knit workgroups where the members had direct connections to each other. Such collaboration was confined to small workgroups and person-to-person communication. Recent developments through the Internet foster virtual workgroups and organizations where dynamic, 'just-in-time' collaboration can take place over a much larger scale. The emergence of virtual workgroups has strongly influenced the interaction of inter-national, inter-disciplinary, as well as educational activities. In this paper we present an array of enabling tools and methods that incorporate the new technologies including web services, software mashups, tag-based structuring and searching, and wikis for collaborative writing and content organization. Large monolithic, 'do-it-all' software tools are giving way to web service modules, combined through service chaining. Application software can now be created using Service Oriented Architecture (SOA). In the air quality community, data providers and users are distributed in space and time creating barriers for data access. By exposing the data on the internet the space, time barriers are lessened. The federated data system, DataFed, developed at Washington University, accesses data from autonomous, distributed providers. Through data "wrappers", DataFed provides uniform and standards-based access services to heterogeneous, distributed data. Service orientation not only lowers the entry resistance for service providers, but it also allows the creation of user-defined applications and/or mashups. For example, Google Earth's open API allowed many groups to mash their content with Google Earth. Ad hoc tagging gives a rich description of the internet resources, but it has the disadvantage of providing a fuzzy schema. The semantic uniformity of the internet resources can be improved by controlled tagging which apply a consistent namespace and tag combinations to diverse objects. One example of this is the photo-sharing web application Flickr. Just like data, by exposing photos through the internet those can be reused in ways unknown and unanticipated by the provider. For air quality application, Flickr allowed a rich collection of images of forest fire smoke, wind blown dust and haze events to be tagged with controlled tags and used in for evaluating subtle features of the events. Wikis, originally used just for collaboratively writing and discuss documents, are now also a social software workflow managers. In air quality data, wikis provides the means to collaboratively create rich metadata. Wikis become a virtual meeting place to discuss ideas before a workshop of conference, display tagged internet resources, and collaboratively work on documents. Wikis are also useful in the classroom. For instance in class projects, the wiki displays harvested resources, maintains collaborative documents and discussions and is the organizational memory for the project.

  3. 75 FR 51823 - Government-Owned Inventions; Availability for Licensing

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-23

    ... applications. Transforming Growth Factor Beta-1 (TGF-[beta]1) Transgenic Mouse Model Description of Technology... developed a transgenic mouse model, designated [beta]1\\glo\\, which permits conditional, gene-specific... gene by Cre recombinase allows expression of TGF-[beta]1. Thus, these mice may be cross-bred with a...

  4. Impact of unusual fatty acid synthesis on futile cycling through beta-oxidation and on gene expression in transgenic plants.

    PubMed

    Moire, Laurence; Rezzonico, Enea; Goepfert, Simon; Poirier, Yves

    2004-01-01

    Arabidopsis expressing the castor bean (Ricinus communis) oleate 12-hydroxylase or the Crepis palaestina linoleate 12-epoxygenase in developing seeds typically accumulate low levels of ricinoleic acid and vernolic acid, respectively. We have examined the presence of a futile cycle of fatty acid degradation in developing seeds using the synthesis of polyhydroxyalkanoate (PHA) from the intermediates of the peroxisomal beta-oxidation cycle. Both the quantity and monomer composition of the PHA synthesized in transgenic plants expressing the 12-epoxygenase and 12-hydroxylase in developing seeds revealed the presence of a futile cycle of degradation of the corresponding unusual fatty acids, indicating a limitation in their stable integration into lipids. The expression profile of nearly 200 genes involved in fatty acid biosynthesis and degradation has been analyzed through microarray. No significant changes in gene expression have been detected as a consequence of the activity of the 12-epoxygenase or the 12-hydroxylase in developing siliques. Similar results have also been obtained for transgenic plants expressing the Cuphea lanceolata caproyl-acyl carrier protein thioesterase and accumulating high amounts of caproic acid. Only in developing siliques of the tag1 mutant, deficient in the accumulation of triacylglycerols and shown to have a substantial futile cycling of fatty acids toward beta-oxidation, have some changes in gene expression been detected, notably the induction of the isocitrate lyase gene. These results indicate that analysis of peroxisomal PHA is a better indicator of the flux of fatty acid through beta-oxidation than the expression profile of genes involved in lipid metabolism.

  5. 77 FR 12240 - Application(s) for Duty-Free Entry of Scientific Instruments

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-29

    ... experiments will consist of direct visual observations of fluorescently tagged DNA and DNA-bound protein... including animal tissues, bacteria, insects and parasites, involving the examination of their morphological...

  6. Utilizing Linked Open Data Sources for Automatic Generation of Semantic Metadata

    NASA Astrophysics Data System (ADS)

    Nummiaho, Antti; Vainikainen, Sari; Melin, Magnus

    In this paper we present an application that can be used to automatically generate semantic metadata for tags given as simple keywords. The application that we have implemented in Java programming language creates the semantic metadata by linking the tags to concepts in different semantic knowledge bases (CrunchBase, DBpedia, Freebase, KOKO, Opencyc, Umbel and/or WordNet). The steps that our application takes in doing so include detecting possible languages, finding spelling suggestions and finding meanings from amongst the proper nouns and common nouns separately. Currently, our application supports English, Finnish and Swedish words, but other languages could be included easily if the required lexical tools (spellcheckers, etc.) are available. The created semantic metadata can be of great use in, e.g., finding and combining similar contents, creating recommendations and targeting advertisements.

  7. The application of PIT tags to measure transport of detrital coral fragments on a fringing reef: Majuro Atoll, Marshall Islands

    NASA Astrophysics Data System (ADS)

    Ford, Murray R.

    2014-06-01

    Passive integrated transponder (PIT) tags are a radio-frequency identification device widely used as a machine-readable identification tool in fisheries research. PIT tags have also been employed, to a lesser extent, to track the movement of gravel-sized clasts within fluvial and coastal systems. In this study, PIT tags were inserted into detrital coral fragments and used to establish source-sink transport pathways on a fringing reef on Majuro Atoll in the Marshall Islands. Results suggest the transport of gravel-sized material on the inter-tidal reef flat is exclusively across-reef towards the lagoon. Considerable variation in the distance travelled by fragments was observed. Fragments were largely intact and visually recognisable after almost 5 months on the reef flat. However, the branches of some recovered fragments had broken off and corallite abrasion was observed in recovered fragments. This study indicates that PIT tags are an inexpensive and powerful new addition to the suite of sediment transport and taphonomic tools for researchers working within coral reef systems.

  8. Automated Data Tagging in the HLA

    NASA Astrophysics Data System (ADS)

    Gaffney, N. I.; Miller, W. W.

    2008-08-01

    One of the more powerful and popular forms of data organization implemented in most popular information sharing web applications is data tagging. With a rich user base from which to gather and digest tags, many interesting and often unanticipated yet very useful associations are revealed. With regard to an existing information, the astronomical community has a rich pool of existing digitally stored and searchable data than any of the currently popular web community, such as You Tube or My Space, had when they started. In initial experiments with the search engine for the Hubble Legacy Archive, we have created a simple yet powerful scheme by which the information from a footprint service, the NED and SIMBAD catalog services, and the ADS abstracts and keywords can be used to initially tag data with standard keywords. By then ingesting this into a public ally available information search engine, such as Apache Lucene, one can create a simple and powerful data tag search engine and association system. By then augmenting this with user provided keys and usage pattern analysis, one can produce a powerful modern data mining system for any astronomical data warehouse.

  9. High-affinity gold nanoparticle pin to label and localize histidine-tagged protein in macromolecular assemblies

    PubMed Central

    Anthony, Kelsey C.; You, Changjiang; Piehler, Jacob; Pomeranz Krummel, Daniel A.

    2014-01-01

    SUMMARY There is significant demand for experimental approaches to aid protein localization in electron microscopy micrographs and ultimately in three-dimensional reconstructions of macromolecular assemblies. We report preparation and use of a reagent consisting of tris-nitrilotriacetic acid (tris-NTA) conjugated with a monofunctional gold nanoparticle (AuNPtris-NTA) for site-specific, non-covalent labeling of protein termini fused to a histidine-tag (His-tag). Multivalent binding of tris-NTA to a His-tag via complexed Ni(II) ions results in subnanomolar affinity and a defined 1:1 stoichiometry. Precise localization of AuNPtris-NTA labeled proteins by electron microscopy is further ensured by the reagent’s short conformationally restricted linker. We have employed AuNPtris-NTA to localize His-tagged proteins in an oligomeric ATPase and in the bacterial 50S ribosomal subunit. AuNPtris-NTA can specifically bind to the target proteins in these assemblies and is clearly discernible. Our new labeling reagent should find broad application in non-covalent site-specific labeling of protein termini to pinpoint their location in macromolecular assemblies. PMID:24560806

  10. Fluorescence from Multiple Chromophore Hydrogen-Bonding States in the Far-Red Protein TagRFP675.

    PubMed

    Konold, Patrick E; Yoon, Eunjin; Lee, Junghwa; Allen, Samantha L; Chapagain, Prem P; Gerstman, Bernard S; Regmi, Chola K; Piatkevich, Kiryl D; Verkhusha, Vladislav V; Joo, Taiha; Jimenez, Ralph

    2016-08-04

    Far-red fluorescent proteins are critical for in vivo imaging applications, but the relative importance of structure versus dynamics in generating large Stokes-shifted emission is unclear. The unusually red-shifted emission of TagRFP675, a derivative of mKate, has been attributed to the multiple hydrogen bonds with the chromophore N-acylimine carbonyl. We characterized TagRFP675 and point mutants designed to perturb these hydrogen bonds with spectrally resolved transient grating and time-resolved fluorescence (TRF) spectroscopies supported by molecular dynamics simulations. TRF results for TagRFP675 and the mKate/M41Q variant show picosecond time scale red-shifts followed by nanosecond time blue-shifts. Global analysis of the TRF spectra reveals spectrally distinct emitting states that do not interconvert during the S1 lifetime. These dynamics originate from photoexcitation of a mixed ground-state population of acylimine hydrogen bond conformers. Strategically tuning the chromophore environment in TagRFP675 might stabilize the most red-shifted conformation and result in a variant with a larger Stokes shift.

  11. Iodine Tagging Velocimetry in a Mach 10 Wake

    NASA Technical Reports Server (NTRS)

    Balla, Robert Jeffrey

    2013-01-01

    A variation on molecular tagging velocimetry (MTV) [1] designated iodine tagging velocimetry (ITV) is demonstrated. Molecular iodine is tagged by two-photon absorption using an Argon Fluoride (ArF) excimer laser. A single camera measures fluid displacement using atomic iodine emission at 206 nm. Two examples ofMTVfor cold-flowmeasurements areN2OMTV [2] and Femtosecond Laser Electronic Excitation Tagging [3]. These, like most MTV methods, are designed for atmospheric pressure applications. Neither can be implemented at the low pressures (0.1- 1 Torr) in typical hypersonic wakes. Of all the single-laser/singlecamera MTV approaches, only Nitric-Oxide Planar Laser Induced Fluorescence-based MTV [4] has been successfully demonstrated in a Mach 10 wake. Oxygen quenching limits transit times to 500 ns and accuracy to typically 30%. The present note describes the photophysics of the ITV method. Off-body velocimetry along a line is demonstrated in the aerothermodynamically important and experimentally challenging region of a hypersonic low-pressure near-wake in a Mach 10 air wind tunnel. Transit times up to 10 µs are demonstrated with conservative errors of 10%.

  12. Twiddlenet: Metadata Tagging and Data Dissemination in Mobile Device Networks

    DTIC Science & Technology

    2007-09-01

    hosting a distributed data dissemination application. Stated simply, there are a multitude of handheld devices on the market that can communicate in...content ( UGC ) across a network of distributed devices. This sharing is accomplished through the use of descriptive metadata tags that are assigned to a...file once it has been shared. These metadata files are uploaded to a centralized portal and arranged for efficient UGC location and searching

  13. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhang, Meng; Cao, Xia; Jia, Qingli

    Triacylglycerol (TAG) is the main storage lipid in plant seeds and the major form of plant oil used for food and, increasingly, for industrial and biofuel applications. Several transcription factors, including FUSCA3 (At3 g26790, FUS3), are associated with embryo maturation and oil biosynthesis in seeds. However, the ability of FUS3 to increase TAG biosynthesis in other tissues has not been quantitatively examined. Here, we evaluated the ability of FUS3 to activate TAG accumulation in non-seed tissues. Overexpression of FUS3 driven by an estradiol-inducible promoter increased oil contents in Arabidopsis seedlings up to 6% of dry weight; more than 50-fold overmore » controls. Eicosenoic acid, a characteristic fatty acid of Arabidopsis seed oil, accumulated to over 20% of fatty acids in cotyledons and leaves. These large increases depended on added sucrose, although without sucrose TAG increased three- to four-fold. Inducing the expression of FUS3 in tobacco BY2 cells also increased TAG accumulation, and co-expression of FUS3 and diacylglycerol acyltransferase 1 (DGAT1) further increased TAG levels to 4% of dry weight. BY2 cell growth was not altered by FUS3 expression, although Arabidopsis seedling development was impaired, consistent with the ability of FUS3 to induce embryo characteristics in non-seed tissues. Microarrays of Arabidopsis seedlings revealed that FUS3 overexpression increased the expression of a higher proportion of genes involved in TAG biosynthesis than genes involved in fatty acid biosynthesis or other lipid pathways. Together these results provide additional insights into FUS3 functions in TAG metabolism and suggest complementary strategies for engineering vegetative oil accumulation.« less

  14. Haplotag: Software for Haplotype-Based Genotyping-by-Sequencing Analysis

    PubMed Central

    Tinker, Nicholas A.; Bekele, Wubishet A.; Hattori, Jiro

    2016-01-01

    Genotyping-by-sequencing (GBS), and related methods, are based on high-throughput short-read sequencing of genomic complexity reductions followed by discovery of single nucleotide polymorphisms (SNPs) within sequence tags. This provides a powerful and economical approach to whole-genome genotyping, facilitating applications in genomics, diversity analysis, and molecular breeding. However, due to the complexity of analyzing large data sets, applications of GBS may require substantial time, expertise, and computational resources. Haplotag, the novel GBS software described here, is freely available, and operates with minimal user-investment on widely available computer platforms. Haplotag is unique in fulfilling the following set of criteria: (1) operates without a reference genome; (2) can be used in a polyploid species; (3) provides a discovery mode, and a production mode; (4) discovers polymorphisms based on a model of tag-level haplotypes within sequenced tags; (5) reports SNPs as well as haplotype-based genotypes; and (6) provides an intuitive visual “passport” for each inferred locus. Haplotag is optimized for use in a self-pollinating plant species. PMID:26818073

  15. Remote monitoring of fish in small streams: A unified approach using PIT tags

    USGS Publications Warehouse

    Zydlewski, G.B.; Horton, G.; Dubreuil, T.; Letcher, B.; Casey, S.; Zydlewski, Joseph D.

    2006-01-01

    Accurate assessments of fish populations are often limited by re-observation or recapture events. Since the early 1990s, passive integrated transponders (PIT tags) have been used to understand the biology of many fish species. Until recently, PIT applications in small streams have been limited to physical recapture events. To maximize recapture probability, we constructed PIT antenna arrays in small streams to remotely detect individual fish. Experiences from two different laboratories (three case studies) allowed us to develop a unified approach to applying PIT technology for enhancing data assessments. Information on equipment, its installation, tag considerations, and array construction is provided. Theoretical and practical definitions are introduced to standardize metrics for assessing detection efficiency. We demonstrate how certain conditions (stream discharge, vibration, and ambient radio frequency noise) affect the detection efficiency and suggest that by monitoring these conditions, expectations of efficiency can be modified. We emphasize the importance of consistently estimating detection efficiency for fisheries applications.

  16. Synthesis and application of a new fluorous-tagged ammonia equivalent.

    PubMed

    Nielsen, Simon D; Smith, Garrick; Begtrup, Mikael; Kristensen, Jesper L

    2010-04-19

    A novel fluorous-tagged ammonia equivalent has been developed. It is based on a nitrogen-oxygen bond, which can be cleaved in a traceless manner by a molybdenum complex or samarium diiodide. The application in the synthesis of ureas, amides, sulfonamides, and carbamates is described. The scope of the fluorous N-O linker is exemplified by the synthesis of itopride, a drug used for the treatment of functional dyspepsia. Itopride was synthesized with the aid of fluorous purification methods and the product was isolated in good overall yield, with high purity. Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Application of whole-body personal TL dosemeters in mixed field beta-gamma radiation.

    PubMed

    Ciupek, K; Aksamit, D; Wołoszczuk, K

    2014-11-01

    Application of whole-body personal TL dosemeters based on a high-sensitivity LiF:Mg,Cu,P (MCP-N) in mixed field beta-gamma radiation has been characterised. The measurements were carried out with (90)Sr/(90)Y, (85)Kr and (137)Cs point sources to calculate the energy response and linearity of the TLD response in a dose range of 0.1-30 mSv. From the result, calibration curves were obtained, enabling the readout of individual dose equivalent Hp(10) from gamma radiation and Hp(0.07) from beta radiation in mixed field beta-gamma. Limitation of the methodology and its application are presented and discussed. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  18. New developments and prospective applications for beta (1,3) glucans.

    PubMed

    Laroche, Celine; Michaud, Philippe

    2007-01-01

    Publications and patents relative to newly observed functions of beta-(1,3)-D-glucans have notably increased in the last few years with the exploitation of their biological activities. The term beta-(1,3)-D-glucans includes a very large number of polysaccharides from bacterial, fungal and vegetable sources. Their structures have a common backbone of beta-(1,3) linked glucopyranosyl residues but the polysaccharidic chain can be beta-(1,6) branched with glucose or integrate some beta-(1,4) linked glucopyranosyl residues in the main chain. Except for the curdlan, a bacterial linear beta-(1,3)-D-glucans, and for the scleroglucan produced by Sclerotium rolfsii, the main drawback limiting the development of these polysaccharides is the lack of efficient processes for their extraction and purification and their cost. However new applications in agronomy, foods, cosmetic and therapeutic could in a next future accentuate the effort of research for their development. So this review focuses on these beta-(1,3)-D-glucans with the objective to detail the strategies employed for their extraction and the relation structure-functions identified when they induce biological activities.

  19. Development of a Short-term Suction-cup Tagging Method for Small Delphinids to Understand the Effects of Climate, Ecosystem, and Anthropogenic Change

    NASA Astrophysics Data System (ADS)

    Pearson, H. C.; Srinivasan, M.

    2016-02-01

    Documented changes in regional abundance and distribution of marine mammals may be driven by climate, ecosystem, and human-induced variations, operating synergistically or individually on different time scales. However, long-term but fine-scale data on animal ranging and foraging patterns are needed to fully understand the mechanism and magnitude of such changes and if/how top predators such as marine mammals are adapting. This is particularly important for dolphins, for which non-invasive, longer duration tags are needed to track their daily and weekly movement patterns in concert with changes in prey. As part of an ongoing study on dusky dolphins (Lagenorhynchus obscurus) in Kaikoura, New Zealand, we are developing a short-term, non-invasive suction-cup tagging method for collecting high resolution data on dolphin foraging and ranging behavior. This is an advancement in the field of animal telemetry as few published studies have tested non-invasive suction-cup tagging methods on small (< 2 m) delphinids, and even fewer report successful deployments. During austral summer 2013-14 and austral winter 2014, we spent 149 h over 31 d searching for and observing dusky dolphins. Of 71 tagging attempts made, 49% (n = 35) were "sticks" (i.e., the tag adhered to the dolphin). The longest tag attachment time was 357 min. Dive depths tended to increase throughout the day, reaching maximum daytime dive depths of c.a. 25 m. This is consistent with dolphin behavior off Kaikoura, as individuals feed mainly at night on mesopelagic organisms. Most (92%, n = 46) dolphins exhibited low-level responses to tagging, indicating this to be an appropriate species on which to continue tagging efforts. Successful trials will facilitate application of the system to other small delphinids. Tagging data collected can be integrated with climate and oceanographic data derived from satellite sensors and other monitoring programs to produce a comprehensive picture of dolphin behavioral ecology.

  20. Tracking animals in freshwater with electronic tags: past, present and future

    USGS Publications Warehouse

    Cooke, Steven J.; Midwood, Jonathan D.; Thiem, Jason D.; Klimley, Peter; Lucas, Martyn C.; Thorstad, Eva B.; Eiler, John; Holbrook, Chris; Ebner, Brendan C.

    2013-01-01

    Considerable technical developments over the past half century have enabled widespread application of electronic tags to the study of animals in the wild, including in freshwater environments. We review the constraints associated with freshwater telemetry and biologging and the technical developments relevant to their use. Technical constraints for tracking animals are often influenced by the characteristics of the animals being studied and the environment they inhabit. Collectively, they influence which and how technologies can be used and their relative effectiveness. Although radio telemetry has historically been the most commonly used technology in freshwater, passive integrated transponder (PIT) technology, acoustic telemetry and biologgers are becoming more popular. Most telemetry studies have focused on fish, although an increasing number have focused on other taxa, such as turtles, crustaceans and molluscs. Key technical developments for freshwater systems include: miniaturization of tags for tracking small-size life stages and species, fixed stations and coded tags for tracking large samples of animals over long distances and large temporal scales, inexpensive PIT systems that enable mass tagging to yield population- and community-level relevant sample sizes, incorporation of sensors into electronic tags, validation of tag attachment procedures with a focus on maintaining animal welfare, incorporation of different techniques (for example, genetics, stable isotopes) and peripheral technologies (for example, geographic information systems, hydroacoustics), development of novel analytical techniques, and extensive international collaboration. Innovations are still needed in tag miniaturization, data analysis and visualization, and in tracking animals over larger spatial scales (for example, pelagic areas of lakes) and in challenging environments (for example, large dynamic floodplain systems, under ice). There seems to be a particular need for adapting various global positioning system and satellite tagging approaches to freshwater. Electronic tagging provides a mechanism to collect detailed information from imperilled animals and species that have no direct economic value. Current and future advances will continue to improve our knowledge of the natural history of aquatic animals and ecological processes in freshwater ecosystems while facilitating evidence-based resource management and conservation.

  1. Identification of a pair of phospholipid:diacylglycerol acyltransferases from developing flax (Linum usitatissimum L.) seed catalyzing the selective production of trilinolenin.

    PubMed

    Pan, Xue; Siloto, Rodrigo M P; Wickramarathna, Aruna D; Mietkiewska, Elzbieta; Weselake, Randall J

    2013-08-16

    The oil from flax (Linum usitatissimum L.) has high amounts of α-linolenic acid (ALA; 18:3(cis)(Δ9,12,15)) and is one of the richest sources of omega-3 polyunsaturated fatty acids (ω-3-PUFAs). To produce ∼57% ALA in triacylglycerol (TAG), it is likely that flax contains enzymes that can efficiently transfer ALA to TAG. To test this hypothesis, we conducted a systematic characterization of TAG-synthesizing enzymes from flax. We identified several genes encoding acyl-CoA:diacylglycerol acyltransferases (DGATs) and phospholipid:diacylglycerol acyltransferases (PDATs) from the flax genome database. Due to recent genome duplication, duplicated gene pairs have been identified for all genes except DGAT2-2. Analysis of gene expression indicated that two DGAT1, two DGAT2, and four PDAT genes were preferentially expressed in flax embryos. Yeast functional analysis showed that DGAT1, DGAT2, and two PDAT enzymes restored TAG synthesis when produced recombinantly in yeast H1246 strain. The activity of particular PDAT enzymes (LuPDAT1 and LuPDAT2) was stimulated by the presence of ALA. Further seed-specific expression of flax genes in Arabidopsis thaliana indicated that DGAT1, PDAT1, and PDAT2 had significant effects on seed oil phenotype. Overall, this study indicated the existence of unique PDAT enzymes from flax that are able to preferentially catalyze the synthesis of TAG containing ALA acyl moieties. The identified LuPDATs may have practical applications for increasing the accumulation of ALA and other polyunsaturated fatty acids in oilseeds for food and industrial applications.

  2. A special repertoire of alpha:beta T cells in neonatal mice.

    PubMed Central

    Bogue, M; Candéias, S; Benoist, C; Mathis, D

    1991-01-01

    According to several functional criteria, the mature thymocytes of neonatal and adult mice are distinctly different. We wondered whether these differences in function might have a structural correlate: do neonates have a distinct repertoire of alpha:beta T cells? In this study, we have exploited the power of polymerase chain reaction technology to generate large numbers of T cell receptor sequences from sorted thymocyte populations from newborn and adult mice. The newborn-derived sequences show very few N nucleotide additions, usually the major source of diversity in T cell receptors. Most interestingly, the paucity of N insertions appears to be exaggerated by selection events that operate during T cell differentiation in the thymus. The significance of these results is largely: (i) that they parallel recent findings on the B cell repertoire in neonates, raising questions about the reactivities specified by such a special repertoire; and (ii) that they suggest a means to 'tag' T cells exported perinatally, allowing one to test the premise that autoreactive T cells derive preferentially from the newborn repertoire. Images PMID:1834457

  3. Hematopoietic Stem Cell Therapy as a Counter-Measure for Human Exploration of Deep Space

    NASA Technical Reports Server (NTRS)

    Ohi, S.; Roach, A.-N.; Ramsahai, S.; Kim, B. C.; Fitzgerald, W.; Riley, D. A.; Gonda, S. R.

    2004-01-01

    Human exploration of deep space depends, in part, on our ability to counter severe/invasive disorders that astronauts experience in space environments. The known symptoms include hematological/cardiac abnormalities,bone and muscle losses, immunodeficiency, neurological disorders, and cancer. Exploiting the extraordinary plasticity of hematopoietic stem cells (HSCs), which differentiate not only to all types of blood cells, but also to various tissues, we have advanced a hypothesis that ome of the space-caused disorders maybe amenable to hematopoietis stem cell therapy(HSCT) so as to maintain promote human exploration of deep space. Using mouse models of human anemia beta-thaiassemia) as well as spaceflight (hindlimb unloading system), we have obtained feasibility results of HSCT for space anemia, muscle loss, and immunodeficiency. For example, in the case of HSCT for muscle loss, the beta-galactosidese marked HSCs were detected in the hindlimbs of unloaded mouse following transplantation by -X-gal wholemaunt staining procedure. Histochemicaland physical analyses indicated structural contribution of HSCs to the muscle. HSCT for immunodeficiency was investigated ising beta-galactosidese gene-tagged Escherichia coli as the infectious agent. Results of the X-gal staining procedure indicated the rapeutic role of the HSCT. To facilitate the HSCT in space, growth of HSCs were optimized in the NASA Rotating Wall Vessel (RWV) culture systems, including Hydrodynamic Focusing Bioreactor (HFB).

  4. Dissipation of contaminants of emerging concern in biosolids applied to non-irrigated farmland in eastern Colorado

    USGS Publications Warehouse

    Yager, Tracy; Furlong, Edward T.; Kolpin, Dana W.; Kinney, Chad A.; Zaugg, Steven D.; Burkhardt, Mark R.

    2013-01-01

    In 2007, a 1.5-year field-scale study was initiated by the U.S. Geological Survey to evaluate the dissipation of contaminants of emerging concern (CECs) following a first agronomic biosolids application to nonirrigated farmland. CECs with the greatest decrease in concentration in the surface biosolids at 180 days post-application included indole, d-limonene, p-cresol, phenol, and skatol. CECs that were present in the largest concentration in 180-day-weathered biosolids included stanols, nonylphenols, bisphenol A, bis(2-ethylhexyl) phthalate, hexahydrohexamethyl cyclopenta-benzopyran (HHCB), and triclosan. CECs that were detected in pre-application soil were 3-beta coprostanol, skatol, acetophenone, beta-sitosterol, beta-stigmastanol, cholesterol, indole, p-cresol, and phenol, most of which are biogenic sterols or fragrances that have natural plant sources in addition to anthropogenic sources, yet their concentrations increased (in some cases, substantially) following biosolids application. Preliminary data indicate the nonylphenols (including NPEO1, NPEO2), OPEO1, benzo[a]pyrene, diethyl phthalate, d-limonene, HHCB, triclosan, and possibly 3-beta coprostanol, skatol, beta-sitosterol, cholesterol, indole, and p-cresol, migrated downward through the soil by 468 days post-application, but indicated little uptake by mature wheat plants. This study indicates that some CECs are sufficiently persistent and mobile to be vertically transported into the soil column following biosolids applications to the land surface, even in semiarid regions.

  5. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chen, K.; Tsai, H.; Decision and Information Sciences

    The technical basis for extending the Model 9977 shipping package periodic maintenance beyond the one-year interval to a maximum of five years is based on the performance of the O-ring seals and the environmental conditions. The DOE Packaging Certification Program (PCP) has tasked Argonne National Laboratory to develop a Radio-Frequency Identification (RFID) temperature monitoring system for use by the facility personnel at DAF/NTS. The RFID temperature monitoring system, depicted in the figure below, consists of the Mk-1 RFId tags, a reader, and a control computer mounted on a mobile platform that can operate as a stand-alone system, or it canmore » be connected to the local IT network. As part of the Conditions of Approval of the CoC, the user must complete the prescribed training to become qualified and be certified for operation of the RFID temperature monitoring system. The training course will be administered by Argonne National Laboratory on behalf of the Headquarters Certifying Official. This is a complete documentation package for the RFID temperature monitoring system of the Model 9977 packagings at NTS. The documentation package will be used for training and certification. The table of contents are: Acceptance Testing Procedure of MK-1 RFID Tags for DOE/EM Nuclear Materials Management Applications; Acceptance Testing Result of MK-1 RFID Tags for DOE/EM Nuclear Materials Management Applications; Performance Test of the Single Bolt Seal Sensor for the Model 9977 Packaging; Calibration of Built-in Thermistors in RFID Tags for Nevada Test Site; Results of Calibration of Built-in Thermistors in RFID Tags; Results of Thermal Calibration of Second Batch of MK-I RFID Tags; Procedure for Installing and Removing MK-1 RFID Tag on Model 9977 Drum; User Guide for RFID Reader and Software for Temperature Monitoring of Model 9977 Drums at NTS; Software Quality Assurance Plan (SQAP) for the ARG-US System; Quality Category for the RFID Temperature Monitoring System; The Documentation Package for the RFID Temperature Monitoring System; Software Test Plan and Results for ARG-US OnSite; Configuration Management Plan (CMP) for the ARG-US System; Requirements Management Plan for the ARG-US System; and Design Management Plan for ARG-US.« less

  6. Primer and platform effects on 16S rRNA tag sequencing

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tremblay, Julien; Singh, Kanwar; Fern, Alison

    Sequencing of 16S rRNA gene tags is a popular method for profiling and comparing microbial communities. The protocols and methods used, however, vary considerably with regard to amplification primers, sequencing primers, sequencing technologies; as well as quality filtering and clustering. How results are affected by these choices, and whether data produced with different protocols can be meaningfully compared, is often unknown. Here we compare results obtained using three different amplification primer sets (targeting V4, V6–V8, and V7–V8) and two sequencing technologies (454 pyrosequencing and Illumina MiSeq) using DNA from a mock community containing a known number of species as wellmore » as complex environmental samples whose PCR-independent profiles were estimated using shotgun sequencing. We find that paired-end MiSeq reads produce higher quality data and enabled the use of more aggressive quality control parameters over 454, resulting in a higher retention rate of high quality reads for downstream data analysis. While primer choice considerably influences quantitative abundance estimations, sequencing platform has relatively minor effects when matched primers are used. In conclusion, beta diversity metrics are surprisingly robust to both primer and sequencing platform biases.« less

  7. Progress towards barium daughter tagging in Xe136 decay using single molecule fluorescence imaging

    NASA Astrophysics Data System (ADS)

    McDonald, Austin; NEXT Collaboration

    2017-09-01

    The existence of Majorana fermions is of great interest as it may be related to the asymmetry between matter and anti-matter particles in the universe. However, the search for them has proven to be a difficult one. Neutrino-less Double Beta decay (NLDB) offers a possible opportunity for direct observation of a Majorana Fermion. The rate for NLDB decay may be as low as 1 count /ton /year if the mass ordering is inverted. Current detector technologies have background rates between 4 to 300 count /ton /year /ROI at the 100kg scale which is much larger than the universal goal of 0.1 count /ton /year /ROI desired for ton-scale detectors. The premise of my research is to develop new detector technologies that will allow for a background-free experiment. My current work is to develop a sensor that will tag the daughter ion Ba++ from the Xe136 decay. The development of a sensor that is sensitive to single barium ion detection based on the single molecule fluorescence imaging technique is the major focus of this work. If successful, this could provide a path to a background-free experiment.

  8. Progress towards barium daughter tagging in Xe136 decay using single molecule fluorescence imaging

    NASA Astrophysics Data System (ADS)

    McDonald, Austin; Jones, Ben; Benson, Jordan; Nygren, David; NEXT Collaboration

    2017-01-01

    The existence of Majorana Fermions has been predicted, and is of great interest as it may be related to the asymmetry between matter and anti-matter particles in the universe. However, the search for them has proven to be a difficult one. Neutrino-less Double Beta decay (NLDB) offers a possible opportunity for direct observation of a Majorana Fermion. The rate for NLDB decay may be as low as 1 count / ton / year . Current detector technologies have background rates between 4 to 300 count / ton / year / ROI which is much larger than the universal goal of 0 . 1 count / ton / year / ROI desired for ton-scale detectors. The premise of my research is to develop new detector technologies that will allow for a background-free experiment. My current work is to develop a sensor that will tag the daughter ion Ba++ from the Xe136 decay. The development of a sensor that is sensitive to single barium ion detection based on the single molecule fluorescence imaging technique is the major focus of this work. If successful, this could provide a path to a background-free experiment.

  9. Primer and platform effects on 16S rRNA tag sequencing

    DOE PAGES

    Tremblay, Julien; Singh, Kanwar; Fern, Alison; ...

    2015-08-04

    Sequencing of 16S rRNA gene tags is a popular method for profiling and comparing microbial communities. The protocols and methods used, however, vary considerably with regard to amplification primers, sequencing primers, sequencing technologies; as well as quality filtering and clustering. How results are affected by these choices, and whether data produced with different protocols can be meaningfully compared, is often unknown. Here we compare results obtained using three different amplification primer sets (targeting V4, V6–V8, and V7–V8) and two sequencing technologies (454 pyrosequencing and Illumina MiSeq) using DNA from a mock community containing a known number of species as wellmore » as complex environmental samples whose PCR-independent profiles were estimated using shotgun sequencing. We find that paired-end MiSeq reads produce higher quality data and enabled the use of more aggressive quality control parameters over 454, resulting in a higher retention rate of high quality reads for downstream data analysis. While primer choice considerably influences quantitative abundance estimations, sequencing platform has relatively minor effects when matched primers are used. In conclusion, beta diversity metrics are surprisingly robust to both primer and sequencing platform biases.« less

  10. High-yield expression of recombinant soybean agglutinin in plants using transient and stable systems.

    PubMed

    Tremblay, Reynald; Feng, Mary; Menassa, Rima; Huner, Norman P A; Jevnikar, Anthony M; Ma, Shengwu

    2011-04-01

    Soybean agglutinin (SBA) is a specific N-acetylgalactosamine-binding plant lectin that can agglutinate a wide variety of cells. SBA has great potential for medical and biotechnology-focused applications, including screening and treatment of breast cancer, isolation of fetal cells from maternal blood for genetic screening, the possibility as a carrier system for oral drug delivery, and utilization as an affinity tag for high-quality purification of tagged proteins. The success of these applications, to a large degree, critically depends on the development of a highly efficient expression system for a source of recombinant SBA (rSBA). Here, we demonstrate the utility of transient and stable expression systems in Nicotiana benthamiana and potato, respectively, for the production of rSBA, with the transgenic protein accumulated to 4% of total soluble protein (TSP) in Nicotiana benthamiana leaves and 0.3% of TSP in potato tubers. Furthermore, we show that both plant-derived rSBAs retain their ability to induce the agglutination of red blood cells, are similarly glycosylated when compared with native SBA, retained their binding specificity for N-acetylgalactosamine, and were highly resistant to degradation in simulated gastric and intestinal fluids. Affinity column purification using N-acetylgalactosamine as a specific ligand resulted in high recovery and purity of rSBA. This work is the first step toward use of rSBA for various new applications, including the development of rSBA as a novel affinity tag for simplified purification of tagged proteins and as a new carrier molecule for delivery of oral drugs.

  11. Cost effective raspberry pi-based radio frequency identification tagging of mice suitable for automated in vivo imaging.

    PubMed

    Bolaños, Federico; LeDue, Jeff M; Murphy, Timothy H

    2017-01-30

    Automation of animal experimentation improves consistency, reduces potential for error while decreasing animal stress and increasing well-being. Radio frequency identification (RFID) tagging can identify individual mice in group housing environments enabling animal-specific tracking of physiological parameters. We describe a simple protocol to radio frequency identification (RFID) tag and detect mice. RFID tags were injected sub-cutaneously after brief isoflurane anesthesia and do not require surgical steps such as suturing or incisions. We employ glass-encapsulated 125kHz tags that can be read within 30.2±2.4mm of the antenna. A raspberry pi single board computer and tag reader enable automated logging and cross platform support is possible through Python. We provide sample software written in Python to provide a flexible and cost effective system for logging the weights of multiple mice in relation to pre-defined targets. The sample software can serve as the basis of any behavioral or physiological task where users will need to identify and track specific animals. Recently, we have applied this system of tagging to automated mouse brain imaging within home-cages. We provide a cost effective solution employing open source software to facilitate adoption in applications such as automated imaging or tracking individual animal weights during tasks where food or water restriction is employed as motivation for a specific behavior. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Sensitive Carbohydrate Detection using Surface Enhanced Raman Tagging

    PubMed Central

    Vangala, Karthikeshwar; Yanney, Michael; Hsiao, Cheng-Te; Wu, Wells W.; Shen, Rong-Fong; Zou, Sige; Sygula, Andrzej; Zhang, Dongmao

    2010-01-01

    Glycomic analysis is an increasingly important field in biological and biomedical research as glycosylation is one of the most important protein post-translational modifications. We have developed a new technique to detect carbohydrates using surface enhanced Raman spectroscopy (SERS) by designing and applying a Rhodamine B derivative as the SERS tag. Using a reductive amination reaction, the Rhodamine-based tag (RT) was successfully conjugated to three model carbohydrates (glucose, lactose and glucuronic acid). SERS detection limits obtained with 632 nm HeNe laser were ~1 nM in concentration for all the RT-carbohydrate conjugates and ~10 fmol in total sample consumption. The dynamic range of the SERS method is about 4 orders of magnitude, spanning from 1 nM to 5 µM. Ratiometric SERS quantification using isotope-substituted SERS internal references also allows comparative quantifications of carbohydrates labeled with RT and deuterium/hydrogen substituted RT tags, respectively. In addition to enhancing the SERS detection of the tagged carbohydrates, the Rhodamine tagging facilitates fluorescence and mass spectrometric detection of carbohydrates. Current fluorescence sensitivity of RT-carbohydrates is ~ 3 nM in concentration while the mass spectrometry (MS) sensitivity is about 1 fmol that was achieved with linear ion trap electrospray ionization (ESI)-MS instrument. Potential applications that take advantage of the high SERS, fluorescence and MS sensitivity of this SERS tagging strategy are discussed for practical glycomic analysis where carbohydrates may be quantified with a fluorescence and SERS technique, and then identified with ESI-MS techniques. PMID:21082777

  13. Intrapulmonary administration of CCL21 gene-modified dendritic cells reduces tumor burden in spontaneous murine bronchoalveolar cell carcinoma.

    PubMed

    Yang, Seok-Chul; Batra, Raj K; Hillinger, Sven; Reckamp, Karen L; Strieter, Robert M; Dubinett, Steven M; Sharma, Sherven

    2006-03-15

    The antitumor efficiency of dendritic cells transduced with an adenovirus vector expressing secondary lymphoid chemokine (CCL21) was evaluated in a murine model of spontaneous bronchoalveolar cell carcinoma. The transgenic mice (CC-10 TAg) express the SV40 large T antigen (TAg) under the Clara cell promoter, develop bilateral, multifocal, and pulmonary adenocarcinomas, and die at 4 months as a result of progressive pulmonary tumor burden. A single intratracheal administration of CCL21 gene-modified dendritic cells (DC-AdCCL21) led to a marked reduction in tumor burden with extensive mononuclear cell infiltration of the tumors. The reduction in tumor burden was accompanied by the enhanced elaboration of type 1 cytokines [IFN-gamma, interleukin (IL)-12, and granulocyte macrophage colony-stimulating factor] and antiangiogenic chemokines (CXCL9 and CXCL10) but a concomitant decrease in the immunosuppressive molecules (IL-10, transforming growth factor-beta, prostaglandin E(2)) in the tumor microenvironment. The DC-AdCCL21 therapy group revealed a significantly greater frequency of tumor-specific T cells releasing IFN-gamma compared with the controls. Continuous therapy with weekly intranasal delivery of DC-AdCCL21 significantly prolonged median survival by >7 weeks in CC-10 TAg mice. Both innate natural killer and specific T-cell antitumor responses significantly increased following DC-AdCCL21 therapy. Significant reduction in tumor burden in a model in which tumors develop in an organ-specific manner provides a strong rationale for further evaluation of intrapulmonary-administered DC-AdCCL21 in regulation of tumor immunity and genetic immunotherapy for lung cancer.

  14. Relative efficacy of casein or soya protein combined with palm or safflower-seed oil on hyperuricaemia in rats.

    PubMed

    Lo, Hui-Chen; Wang, Yao-Horng; Chiou, Hue-Ying; Lai, Shan-Hu; Yang, Yu

    2010-07-01

    Diets that ameliorate the adverse effects of uric acid (UA) on renal damage deserve attention. The effects of casein or soya protein combined with palm or safflower-seed oil on various serum parameters and renal histology were investigated on hyperuricaemic rats. Male Wistar rats administered with oxonic acid and UA to induce hyperuricaemia were fed with casein or soya protein plus palm- or safflower-seed oil-supplemented diets. Normal rats and hyperuricaemic rats with or without allopurinol treatment (150 mg/l in drinking water) were fed with casein plus maize oil-supplemented diets. After 8 weeks, allopurinol treatment and soya protein plus safflower-seed oil-supplemented diet significantly decreased serum UA in hyperuricaemic rats (one-way ANOVA; P < 0.05). In addition, soya protein and casein attenuated hyperuricaemia-induced decreases in serum albumin and insulin, respectively (two-way ANOVA; P < 0.05). Safflower-seed oil significantly decreased serum TAG and UA, whereas palm oil significantly increased serum cholesterol, TAG, blood urea N and creatinine. However, soya protein significantly decreased renal NO and nitrotyrosine and palm oil significantly decreased renal nitrotyrosine, TNF-alpha and interferon-gamma and increased renal transforming growth factor-beta. Casein with safflower-seed oil significantly attenuated renal tubulointerstitial nephritis, crystals and fibrosis. Comparing casein v. soya protein combined with palm or safflower-seed oil, the results support that casein with safflower-seed oil may be effective in attenuating hyperuricaemia-associated renal damage, while soya protein with safflower-seed oil may be beneficial in lowering serum UA and TAG.

  15. The structure of the SBP-Tag–streptavidin complex reveals a novel helical scaffold bridging binding pockets on separate subunits

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Barrette-Ng, Isabelle H.; Wu, Sau-Ching; Tjia, Wai-Mui

    2013-05-01

    The structure of the SBP-Tag–streptavidin complex reveals a novel mode of peptide recognition in which a single peptide binds simultaneously to biotin-binding pockets from adjacent subunits of streptavidin. The molecular details of peptide recognition suggest how the SBP-Tag can be further modified to become an even more useful tag for a wider range of biotechnological applications. The 38-residue SBP-Tag binds to streptavidin more tightly (K{sub d} ≃ 2.5–4.9 nM) than most if not all other known peptide sequences. Crystallographic analysis at 1.75 Å resolution shows that the SBP-Tag binds to streptavidin in an unprecedented manner by simultaneously interacting with biotin-bindingmore » pockets from two separate subunits. An N-terminal HVV peptide sequence (residues 12–14) and a C-terminal HPQ sequence (residues 31–33) form the bulk of the direct interactions between the SBP-Tag and the two biotin-binding pockets. Surprisingly, most of the peptide spanning these two sites (residues 17–28) adopts a regular α-helical structure that projects three leucine side chains into a groove formed at the interface between two streptavidin protomers. The crystal structure shows that residues 1–10 and 35–38 of the original SBP-Tag identified through in vitro selection and deletion analysis do not appear to contact streptavidin and thus may not be important for binding. A 25-residue peptide comprising residues 11–34 (SBP-Tag2) was synthesized and shown using surface plasmon resonance to bind streptavidin with very similar affinity and kinetics when compared with the SBP-Tag. The SBP-Tag2 was also added to the C-terminus of β-lactamase and was shown to be just as effective as the full-length SBP-Tag in affinity purification. These results validate the molecular structure of the SBP-Tag–streptavidin complex and establish a minimal bivalent streptavidin-binding tag from which further rational design and optimization can proceed.« less

  16. Beta-phenylethylamine stimulates striatal acetylcholine release through activation of the AMPA glutamatergic pathway.

    PubMed

    Ishida, Kota; Murata, Mikio; Kato, Masatoshi; Utsunomiya, Iku; Hoshi, Keiko; Taguchi, Kyoji

    2005-09-01

    Using an in vivo intra-striatal microdialysis technique, we examined the effects of systemically administered beta-phenylethylamine (beta-PEA), a psychomotor stimulating trace amine, on striatal acetylcholine release in freely moving rats. Infusion of N-methyl-D-aspartic acid (NMDA; 10(-5) M) significantly increased acetylcholine release. In addition, locally applied amino-3-hydroxy-5-methylisozasole-4-propionic acid (AMPA; 10(-5) M) significantly increased acetylcholine release in the striatum. Intra-striatal application of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10(-5) M), an AMPA-type glutamatergic receptor antagonist, had little effect on acetylcholine release, while application of MK-801 (10(-5) M, 10(-6) M), an NMDA-type glutamatergic receptor antagonist, significantly reduced acetylcholine release. Acetylcholine within striatal perfusate was significantly increased by intraperitoneal administration of beta-PEA in a dose-dependent manner. This increase in acetylcholine release was completely blocked by application of CNQX (10(-5) M) through the microdialysis probe into the striatum. However, increased acetylcholine response to systemic beta-PEA was unaltered by addition of MK-801 to the perfusion medium. These results suggest a regulatory function of beta-PEA, mediated by AMPA-type glutamatergic receptors, on the release of acetylcholine in the rat striatum.

  17. Advancement Of Tritium Powered Betavoltaic Battery Systems

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Staack, G.; Gaillard, J.; Hitchcock, D.

    2015-10-14

    Due to their decades-long service life and reliable power output under extreme conditions, betavoltaic batteries offer distinct advantages over traditional chemical batteries, especially in applications where frequent battery replacement is hazardous, or cost prohibitive. Although many beta emitting isotopes exist, tritium is considered ideal in betavoltaic applications for several reasons: 1) it is a “pure” beta emitter, 2) the beta is not energetic enough to damage the semiconductor, 3) it has a moderately long half-life, and 4) it is readily available. Unfortunately, the widespread application of tritium powered betavoltaics is limited, in part, by their low power output. This researchmore » targets improving the power output of betavoltaics by increasing the flux of beta particles to the energy conversion device (the p-n junction) through the use of low Z nanostructured tritium trapping materials.« less

  18. The Cation−π Interaction Enables a Halo-Tag Fluorogenic Probe for Fast No-Wash Live Cell Imaging and Gel-Free Protein Quantification

    PubMed Central

    2017-01-01

    The design of fluorogenic probes for a Halo tag is highly desirable but challenging. Previous work achieved this goal by controlling the chemical switch of spirolactones upon the covalent conjugation between the Halo tag and probes or by incorporating a “channel dye” into the substrate binding tunnel of the Halo tag. In this work, we have developed a novel class of Halo-tag fluorogenic probes that are derived from solvatochromic fluorophores. The optimal probe, harboring a benzothiadiazole scaffold, exhibits a 1000-fold fluorescence enhancement upon reaction with the Halo tag. Structural, computational, and biochemical studies reveal that the benzene ring of a tryptophan residue engages in a cation−π interaction with the dimethylamino electron-donating group of the benzothiadiazole fluorophore in its excited state. We further demonstrate using noncanonical fluorinated tryptophan that the cation−π interaction directly contributes to the fluorogenicity of the benzothiadiazole fluorophore. Mechanistically, this interaction could contribute to the fluorogenicity by promoting the excited-state charge separation and inhibiting the twisting motion of the dimethylamino group, both leading to an enhanced fluorogenicity. Finally, we demonstrate the utility of the probe in no-wash direct imaging of Halo-tagged proteins in live cells. In addition, the fluorogenic nature of the probe enables a gel-free quantification of fusion proteins expressed in mammalian cells, an application that was not possible with previously nonfluorogenic Halo-tag probes. The unique mechanism revealed by this work suggests that incorporation of an excited-state cation−π interaction could be a feasible strategy for enhancing the optical performance of fluorophores and fluorogenic sensors. PMID:28221782

  19. The Cation-π Interaction Enables a Halo-Tag Fluorogenic Probe for Fast No-Wash Live Cell Imaging and Gel-Free Protein Quantification.

    PubMed

    Liu, Yu; Miao, Kun; Dunham, Noah P; Liu, Hongbin; Fares, Matthew; Boal, Amie K; Li, Xiaosong; Zhang, Xin

    2017-03-21

    The design of fluorogenic probes for a Halo tag is highly desirable but challenging. Previous work achieved this goal by controlling the chemical switch of spirolactones upon the covalent conjugation between the Halo tag and probes or by incorporating a "channel dye" into the substrate binding tunnel of the Halo tag. In this work, we have developed a novel class of Halo-tag fluorogenic probes that are derived from solvatochromic fluorophores. The optimal probe, harboring a benzothiadiazole scaffold, exhibits a 1000-fold fluorescence enhancement upon reaction with the Halo tag. Structural, computational, and biochemical studies reveal that the benzene ring of a tryptophan residue engages in a cation-π interaction with the dimethylamino electron-donating group of the benzothiadiazole fluorophore in its excited state. We further demonstrate using noncanonical fluorinated tryptophan that the cation-π interaction directly contributes to the fluorogenicity of the benzothiadiazole fluorophore. Mechanistically, this interaction could contribute to the fluorogenicity by promoting the excited-state charge separation and inhibiting the twisting motion of the dimethylamino group, both leading to an enhanced fluorogenicity. Finally, we demonstrate the utility of the probe in no-wash direct imaging of Halo-tagged proteins in live cells. In addition, the fluorogenic nature of the probe enables a gel-free quantification of fusion proteins expressed in mammalian cells, an application that was not possible with previously nonfluorogenic Halo-tag probes. The unique mechanism revealed by this work suggests that incorporation of an excited-state cation-π interaction could be a feasible strategy for enhancing the optical performance of fluorophores and fluorogenic sensors.

  20. Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli.

    PubMed

    Lázaro, Beatriz; Villa, Juan A; Santín, Omar; Cabezas, Matilde; Milagre, Cintia D F; de la Cruz, Fernando; Moncalián, Gabriel

    2017-01-01

    Triglycerides (TAGs), the major storage molecules of metabolic energy and source of fatty acids, are produced as single cell oil by some oleogenic microorganisms. However, these microorganisms require strict culture conditions, show low carbon source flexibilities, lack efficient genetic modification tools and in some cases pose safety concerns. TAGs have essential applications such as behaving as a source for added-value fatty acids or giving rise to the production of biodiesel. Hence, new alternative methods are urgently required for obtaining these oils. In this work we describe TAG accumulation in the industrially appropriate microorganism Escherichia coli expressing the heterologous enzyme tDGAT, a wax ester synthase/triacylglycerol:acylCoA acyltranferase (WS/DGAT). With this purpose, we introduce a codon-optimized gene from the thermophilic actinomycete Thermomonospora curvata coding for a WS/DGAT into different E. coli strains, describe the metabolic effects associated to the expression of this protein and evaluate neutral lipid accumulation. We observe a direct relation between the expression of this WS/DGAT and TAG production within a wide range of culture conditions. More than 30% TAGs were detected within the bacterial neutral lipids in 90 minutes after induction. TAGs were observed to be associated with the hydrophobic enzyme while forming round intracytoplasmic bodies, which could represent a bottleneck for lipid accumulation in E. coli. We detected an increase of almost 3-fold in the monounsaturated fatty acids (MUFA) occurring in the recombinant strains. These MUFA were predominant in the accumulated TAGs achieving 46% of the TAG fatty acids. These results set the basis for further research on the achievement of a suitable method towards the sustainable production of these neutral lipids.

  1. FUSCA3 activates triacylglycerol accumulation in Arabidopsis seedlings and tobacco BY2 cells

    DOE PAGES

    Zhang, Meng; Cao, Xia; Jia, Qingli; ...

    2016-06-11

    Triacylglycerol (TAG) is the main storage lipid in plant seeds and the major form of plant oil used for food and, increasingly, for industrial and biofuel applications. Several transcription factors, including FUSCA3 (At3 g26790, FUS3), are associated with embryo maturation and oil biosynthesis in seeds. However, the ability of FUS3 to increase TAG biosynthesis in other tissues has not been quantitatively examined. Here, we evaluated the ability of FUS3 to activate TAG accumulation in non-seed tissues. Overexpression of FUS3 driven by an estradiol-inducible promoter increased oil contents in Arabidopsis seedlings up to 6% of dry weight; more than 50-fold overmore » controls. Eicosenoic acid, a characteristic fatty acid of Arabidopsis seed oil, accumulated to over 20% of fatty acids in cotyledons and leaves. These large increases depended on added sucrose, although without sucrose TAG increased three- to four-fold. Inducing the expression of FUS3 in tobacco BY2 cells also increased TAG accumulation, and co-expression of FUS3 and diacylglycerol acyltransferase 1 (DGAT1) further increased TAG levels to 4% of dry weight. BY2 cell growth was not altered by FUS3 expression, although Arabidopsis seedling development was impaired, consistent with the ability of FUS3 to induce embryo characteristics in non-seed tissues. Microarrays of Arabidopsis seedlings revealed that FUS3 overexpression increased the expression of a higher proportion of genes involved in TAG biosynthesis than genes involved in fatty acid biosynthesis or other lipid pathways. Together these results provide additional insights into FUS3 functions in TAG metabolism and suggest complementary strategies for engineering vegetative oil accumulation.« less

  2. Application of volcanic ash particles for protein affinity purification with a minimized silica-binding tag.

    PubMed

    Abdelhamid, Mohamed A A; Ikeda, Takeshi; Motomura, Kei; Tanaka, Tatsuya; Ishida, Takenori; Hirota, Ryuichi; Kuroda, Akio

    2016-11-01

    We recently reported that the spore coat protein, CotB1 (171 amino acids), from Bacillus cereus mediates silica biomineralization and that the polycationic C-terminal sequence of CotB1 (14 amino acids), designated CotB1p, serves as a silica-binding tag when fused to other proteins. Here, we reduced the length of this silica-binding tag to only seven amino acids (SB7 tag: RQSSRGR) while retaining its affinity for silica. Alanine scanning mutagenesis indicated that the three arginine residues in the SB7 tag play important roles in binding to a silica surface. Monomeric l-arginine, at concentrations of 0.3-0.5 M, was found to serve as a competitive eluent to release bound SB7-tagged proteins from silica surfaces. To develop a low-cost, silica-based affinity purification procedure, we used natural volcanic ash particles with a silica content of ∼70%, rather than pure synthetic silica particles, as an adsorbent for SB7-tagged proteins. Using green fluorescent protein, mCherry, and mKate2 as model proteins, our purification method achieved 75-90% recovery with ∼90% purity. These values are comparable to or even higher than that of the commonly used His-tag affinity purification. In addition to low cost, another advantage of our method is the use of l-arginine as the eluent because its protein-stabilizing effect would help minimize alteration of the intrinsic properties of the purified proteins. Our approach paves the way for the use of naturally occurring materials as adsorbents for simple, low-cost affinity purification. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  3. FUSCA3 activates triacylglycerol accumulation in Arabidopsis seedlings and tobacco BY2 cells.

    PubMed

    Zhang, Meng; Cao, Xia; Jia, Qingli; Ohlrogge, John

    2016-10-01

    Triacylglycerol (TAG) is the main storage lipid in plant seeds and the major form of plant oil used for food and, increasingly, for industrial and biofuel applications. Several transcription factors, including FUSCA3 (At3 g26790, FUS3), are associated with embryo maturation and oil biosynthesis in seeds. However, the ability of FUS3 to increase TAG biosynthesis in other tissues has not been quantitatively examined. Here, we evaluated the ability of FUS3 to activate TAG accumulation in non-seed tissues. Overexpression of FUS3 driven by an estradiol-inducible promoter increased oil contents in Arabidopsis seedlings up to 6% of dry weight; more than 50-fold over controls. Eicosenoic acid, a characteristic fatty acid of Arabidopsis seed oil, accumulated to over 20% of fatty acids in cotyledons and leaves. These large increases depended on added sucrose, although without sucrose TAG increased three- to four-fold. Inducing the expression of FUS3 in tobacco BY2 cells also increased TAG accumulation, and co-expression of FUS3 and diacylglycerol acyltransferase 1 (DGAT1) further increased TAG levels to 4% of dry weight. BY2 cell growth was not altered by FUS3 expression, although Arabidopsis seedling development was impaired, consistent with the ability of FUS3 to induce embryo characteristics in non-seed tissues. Microarrays of Arabidopsis seedlings revealed that FUS3 overexpression increased the expression of a higher proportion of genes involved in TAG biosynthesis than genes involved in fatty acid biosynthesis or other lipid pathways. Together these results provide additional insights into FUS3 functions in TAG metabolism and suggest complementary strategies for engineering vegetative oil accumulation. © 2016 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.

  4. Metabolic engineering of sugarcane to accumulate energy-dense triacylglycerols in vegetative biomass

    DOE PAGES

    Zale, Janice; Jung, Je Hyeong; Kim, Jae Yoon; ...

    2015-06-08

    Elevating the lipid content in vegetative tissues has emerged as a new strategy for increasing energy density and biofuel yield of crops. Storage lipids in contrast to structural and signaling lipids are mainly composed of glycerol esters of fatty acids, also known as triacylglycerol (TAG). TAGs are one of the most energy-rich and abundant forms of reduced carbon available in nature. Therefore, altering the carbon-partitioning balance in favour of TAG in vegetative tissues of sugarcane, one of the highest yielding biomass crops, is expected to drastically increase energy yields. We report metabolic engineering to elevate TAG accumulation in vegetative tissuesmore » of sugarcane. Constitutive co-expression of WRINKLED1 (WRI1), diacylglycerol acyltransferase1-2 (DGAT1-2) and oleosin1 (OLE1) and simultaneous cosuppression of ADP-glucose pyrophosphorylase (AGPase) and a subunit of the peroxisomal ABC transporter1 (PXA1) in transgenic sugarcane elevated TAG accumulation in leaves or stems by 95- or 43-fold to 1.9% or 0.9% of dry weight (DW), respectively, while expression or suppression of one to three of the target genes increased TAG levels by 1.5- to 9.5-fold. Accumulation of TAG in vegetative progeny plants was consistent with the results from primary transgenics and contributed to a total fatty acid content of up to 4.7% or 1.7% of DW in mature leaves or stems, respectively. We saw lipid droplets in mesophyll cells of transgenic leaves by confocal fluorescence microscopy. These results provide the basis for optimizations of TAG accumulation in sugarcane and other high yielding biomass grasses and will open new prospects for biofuel applications.« less

  5. Base induced chemical conversion of 3-carbamoyl-2-isoxazolines.

    PubMed

    Nishiwaki, Nagatoshi; Maki, Asaka; Ariga, Masahiro

    2009-01-01

    3-Carbamoyl-2-isoxazolines, prepared by cycloaddition of functionalized nitrile oxide, serve as masked 3-unsubstituted isoxazolines to afford 2-isoxazoline-3-carboxylic acid, beta-cyanoalcohol, alpha,beta-unsaturated nitrile, and alpha,beta-unsaturated amide upon heating in the alkaline solution. The present reaction is also applicable to synthesis of 3,4-difunctionalized isoxazoles and beta-hydroxy-gamma-lactone.

  6. Transforming growth factor- 1 C-509T polymorphism, oxidant stress, and early-onset childhood asthma.

    PubMed

    Salam, Muhammad T; Gauderman, W James; McConnell, Rob; Lin, Pi-Chu; Gilliland, Frank D

    2007-12-15

    Transforming growth factor (TGF)-beta1 is involved in airway inflammation and remodeling, two key processes in asthma pathogenesis. Tobacco smoke and traffic emissions induce airway inflammation and modulate TGF-beta1 gene expression. We hypothesized that the effects of functional TGF-beta1 variants on asthma occurrence vary by these exposures. We tested these hypotheses among 3,023 children who participated in the Children's Health Study. Tagging single-nucleotide polymorphisms rs4803457 C>T and C-509T (a functional promoter polymorphism) accounted for 94% of the haplotype diversity of the upstream region. Exposure to maternal smoking in utero was based on smoking by biological mother during pregnancy. Residential distance from nearest freeway was calculated based on residential address at study entry. Children with the -509TT genotype had a 1.8-fold increased risk of early persistent asthma (95% confidence interval [CI], 1.11-2.95). This association varied marginally significantly by in utero exposure to maternal smoking. Compared with children with the -509CC/CT genotype with no in utero exposure to maternal smoking, those with the -509TT genotype with such exposure had a 3.4-fold increased risk of early persistent asthma (95% CI, 1.46-7.80; interaction, P = 0.11). The association between TGF-beta1 C-509T and lifetime asthma varied by residential proximity to freeways (interaction P = 0.02). Children with the -509TT genotype living within 500 m of a freeway had over three-fold increased lifetime asthma risk (95% CI, 1.29-7.44) compared with children with CC/CT genotype living > 1500 m from a freeway. Children with the TGF-beta1 -509TT genotype are at increased risk of asthma when they are exposed to maternal smoking in utero or to traffic-related emissions.

  7. 50 CFR 697.6 - Dealer permits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... requirement; date(s) of purchases and receipts; units of measure and amount by species (by market category, if applicable); price per unit by species (by market category, if applicable) or total value by species (by market category, if applicable); port landed; cage tag numbers for surfclams and ocean quahogs, if...

  8. 50 CFR 648.7 - Recordkeeping and reporting requirements.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...; date(s) of purchases and receipts; units of measure and amount by species (by market category, if applicable); price per unit by species (by market category, if applicable) or total value by species (by market category, if applicable); port landed; cage tag numbers for surfclams and ocean quahogs, if...

  9. 50 CFR 648.7 - Recordkeeping and reporting requirements.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...; date(s) of purchases and receipts; units of measure and amount by species (by market category, if applicable); price per unit by species (by market category, if applicable) or total value by species (by market category, if applicable); port landed; cage tag numbers for surfclams and ocean quahogs, if...

  10. 50 CFR 648.7 - Recordkeeping and reporting requirements.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...; date(s) of purchases and receipts; units of measure and amount by species (by market category, if applicable); price per unit by species (by market category, if applicable) or total value by species (by market category, if applicable); port landed; cage tag numbers for surfclams and ocean quahogs, if...

  11. 50 CFR 697.6 - Dealer permits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... requirement; date(s) of purchases and receipts; units of measure and amount by species (by market category, if applicable); price per unit by species (by market category, if applicable) or total value by species (by market category, if applicable); port landed; cage tag numbers for surfclams and ocean quahogs, if...

  12. 50 CFR 648.7 - Recordkeeping and reporting requirements.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...; date(s) of purchases and receipts; units of measure and amount by species (by market category, if applicable); price per unit by species (by market category, if applicable) or total value by species (by market category, if applicable); port landed; cage tag numbers for surfclams and ocean quahogs, if...

  13. 50 CFR 697.6 - Dealer permits.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... requirement; date(s) of purchases and receipts; units of measure and amount by species (by market category, if applicable); price per unit by species (by market category, if applicable) or total value by species (by market category, if applicable); port landed; cage tag numbers for surfclams and ocean quahogs, if...

  14. A beta-galactosidase gene is expressed during mature fruit abscission of 'Valencia' orange (Citrus sinensis).

    PubMed

    Wu, Zhencai; Burns, Jacqueline K

    2004-07-01

    beta-galactosidases have been detected in a wide range of plants and are characterized by their ability to hydrolyse terminal non-reducing beta-D-galactosyl residues from beta-D-galactosides. These enzymes have been detected in a wide range of plant organs and tissues. In a search for differentially expressed genes during the abscission process in citrus, sequences encoding beta-galactosidase were identified. Three cDNA fragments of a beta-galactosidase gene were isolated from a cDNA subtraction library constructed from mature fruit abscission zones 48 h after the application of a mature fruit-specific abscission agent, 5-chloro-3-methyl-4-nitro-1H-pyrazole (CMN-pyrazole). Based on sequence information derived from these fragments, a full-length cDNA of 2847 nucleotides (GenBank accession number AY029198) encoding beta-galactosidase was isolated from mature fruit abscission zones by 5'- and 3'-RACE approaches. The beta-galactosidase cDNA encoded a protein of 737 amino acid residues with a calculated molecular weight of 82 kDa. The deduced protein was highly homologous to plant beta-galactosidases expressed in fruit ripening. Southern blot analysis demonstrated that at least two closely related beta-galactosidase genes were present in 'Valencia' orange. Temporal expression patterns in mature fruit abscission zones indicated beta-galactosidase mRNA was detected 48 h after treatment of CMN-pyrazole and ethephon in mature fruit abscission zones. beta-galactosidase transcripts were detected in leaf abscission zones only after ethephon application. The citrus beta-galactosidase was expressed in stamens and petals of fully opened flowers and young fruitlets. The results suggest that this beta-galactosidase may play a role during abscission as well as early growth and development processes in flowers and fruitlets.

  15. Production of fatty acid components of meadowfoam oil in somatic soybean embryos.

    PubMed

    Cahoon, E B; Marillia, E F; Stecca, K L; Hall, S E; Taylor, D C; Kinney, A J

    2000-09-01

    The seed oil of meadowfoam (Limnanthes alba) and other Limnanthes spp. is enriched in the unusual fatty acid Delta(5)-eicosenoic acid (20:1Delta(5)). This fatty acid has physical and chemical properties that make the seed oil of these plants useful for a number of industrial applications. An expressed sequence tag approach was used to identify cDNAs for enzymes involved in the biosynthesis of 20:1Delta(5)). By random sequencing of a library prepared from developing Limnanthes douglasii seeds, a class of cDNAs was identified that encode a homolog of acyl-coenzyme A (CoA) desaturases found in animals, fungi, and cyanobacteria. Expression of a cDNA for the L. douglasii acyl-CoA desaturase homolog in somatic soybean (Glycine max) embryos behind a strong seed-specific promoter resulted in the accumulation of Delta(5)-hexadecenoic acid to amounts of 2% to 3% (w/w) of the total fatty acids of single embryos. Delta(5)-Octadecenoic acid and 20:1Delta(5) also composed <1% (w/w) each of the total fatty acids of these embryos. In addition, cDNAs were identified from the L. douglasii expressed sequence tags that encode a homolog of fatty acid elongase 1 (FAE1), a beta-ketoacyl-CoA synthase that catalyzes the initial step of very long-chain fatty acid synthesis. Expression of the L. douglassi FAE1 homolog in somatic soybean embryos was accompanied by the accumulation of C(20) and C(22) fatty acids, principally as eicosanoic acid, to amounts of 18% (w/w) of the total fatty acids of single embryos. To partially reconstruct the biosynthetic pathway of 20:1Delta(5) in transgenic plant tissues, cDNAs for the L. douglasii acyl-CoA desaturase and FAE1 were co-expressed in somatic soybean embryos. In the resulting transgenic embryos, 20:1Delta(5) and Delta(5)-docosenoic acid composed up to 12% of the total fatty acids.

  16. Advances in chemical labeling of proteins in living cells.

    PubMed

    Yan, Qi; Bruchez, Marcel P

    2015-04-01

    The pursuit of quantitative biological information via imaging requires robust labeling approaches that can be used in multiple applications and with a variety of detectable colors and properties. In addition to conventional fluorescent proteins, chemists and biologists have come together to provide a range of approaches that combine dye chemistry with the convenience of genetic targeting. This hybrid-tagging approach amalgamates the rational design of properties available through synthetic dye chemistry with the robust biological targeting available with genetic encoding. In this review, we discuss the current range of approaches that have been exploited for dye targeting or for targeting and activation and some of the recent applications that are uniquely permitted by these hybrid-tagging approaches.

  17. Design of Inkjet-Printed RFID-Based Sensor on Paper: Single- and Dual-Tag Sensor Topologies.

    PubMed

    Kim, Sangkil; Georgiadis, Apostolos; Tentzeris, Manos M

    2018-06-17

    The detailed design considerations for the printed RFID-based sensor system is presented in this paper. Starting from material selection and metallization method, this paper discusses types of RFID-based sensors (single- & dual-tag sensor topologies), design procedures, and performance evaluation methods for the wireless sensor system. The electrical properties of the paper substrates (cellulose-based and synthetic papers) and the silver nano-particle-based conductive film are thoroughly characterized for RF applications up to 8 GHz. The reported technology could potentially set the foundation for truly “green”, low-cost, scalable wireless topologies for autonomous Internet-of-Things (IoT), bio-monitoring, and “smart skin” applications.

  18. An introduction to mass cytometry: fundamentals and applications.

    PubMed

    Tanner, Scott D; Baranov, Vladimir I; Ornatsky, Olga I; Bandura, Dmitry R; George, Thaddeus C

    2013-05-01

    Mass cytometry addresses the analytical challenges of polychromatic flow cytometry by using metal atoms as tags rather than fluorophores and atomic mass spectrometry as the detector rather than photon optics. The many available enriched stable isotopes of the transition elements can provide up to 100 distinguishable reporting tags, which can be measured simultaneously because of the essential independence of detection provided by the mass spectrometer. We discuss the adaptation of traditional inductively coupled plasma mass spectrometry to cytometry applications. We focus on the generation of cytometry-compatible data and on approaches to unsupervised multivariate clustering analysis. Finally, we provide a high-level review of some recent benchmark reports that highlight the potential for massively multi-parameter mass cytometry.

  19. Vehicle Tracking System using Nanotechnology Satellites and Tags

    NASA Technical Reports Server (NTRS)

    Lorenzini, Dino A.; Tubis, Chris

    1995-01-01

    This paper describes a joint project to design, develop, and deploy a satellite based tracking system incorporating micro-nanotechnology components. The system consists of a constellation of 'nanosats', a satellite command station and data collection sites, and a large number of low-cost electronic 'tags'. Both government and commercial applications are envisioned for the satellite based tracking system. The projected low price for the tracking service is made possible by the lightweight nanosats and inexpensive electronic tags which use high production volume single chip transceivers and microprocessor devices. The nanosat consists of a five inch aluminum cube with body mounted solar panels (GaAs solar cells) on all six faces. A UHF turnstile antenna and a simple, spring release mechanism complete the external configuration of the spacecraft.

  20. Multi-Scale Distributed Representation for Deep Learning and its Application to b-Jet Tagging

    NASA Astrophysics Data System (ADS)

    Lee, Jason Sang Hun; Park, Inkyu; Park, Sangnam

    2018-06-01

    Recently machine learning algorithms based on deep layered artificial neural networks (DNNs) have been applied to a wide variety of high energy physics problems such as jet tagging or event classification. We explore a simple but effective preprocessing step which transforms each realvalued observational quantity or input feature into a binary number with a fixed number of digits. Each binary digit represents the quantity or magnitude in different scales. We have shown that this approach improves the performance of DNNs significantly for some specific tasks without any further complication in feature engineering. We apply this multi-scale distributed binary representation to deep learning on b-jet tagging using daughter particles' momenta and vertex information.

  1. Image Location Estimation by Salient Region Matching.

    PubMed

    Qian, Xueming; Zhao, Yisi; Han, Junwei

    2015-11-01

    Nowadays, locations of images have been widely used in many application scenarios for large geo-tagged image corpora. As to images which are not geographically tagged, we estimate their locations with the help of the large geo-tagged image set by content-based image retrieval. In this paper, we exploit spatial information of useful visual words to improve image location estimation (or content-based image retrieval performances). We proposed to generate visual word groups by mean-shift clustering. To improve the retrieval performance, spatial constraint is utilized to code the relative position of visual words. We proposed to generate a position descriptor for each visual word and build fast indexing structure for visual word groups. Experiments show the effectiveness of our proposed approach.

  2. Correlative light and electron microscopic detection of GFP-labeled proteins using modular APEX.

    PubMed

    Ariotti, Nicholas; Hall, Thomas E; Parton, Robert G

    2017-01-01

    The use of green fluorescent protein (GFP) and related proteins has revolutionized light microscopy. Here we describe a rapid and simple method to localize GFP-tagged proteins in cells and in tissues by electron microscopy (EM) using a modular approach involving a small GFP-binding peptide (GBP) fused to the ascorbate peroxidase-derived APEX2 tag. We provide a method for visualizing GFP-tagged proteins by light and EM in cultured cells and in the zebrafish using modular APEX-GBP. Furthermore, we describe in detail the benefits of this technique over many of the currently available correlative light and electron microscopy approaches and demonstrate APEX-GBP is readily applicable to modern three-dimensional techniques. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. PATL: A RFID Tag Localization based on Phased Array Antenna.

    PubMed

    Qiu, Lanxin; Liang, Xiaoxuan; Huang, Zhangqin

    2017-03-15

    In RFID systems, how to detect the position precisely is an important and challenging research topic. In this paper, we propose a range-free 2D tag localization method based on phased array antenna, called PATL. This method takes advantage of the adjustable radiation angle of the phased array antenna to scan the surveillance region in turns. By using the statistics of the tags' number in different antenna beam directions, a weighting algorithm is used to calculate the position of the tag. This method can be applied to real-time location of multiple targets without usage of any reference tags or additional readers. Additionally, we present an optimized weighting method based on RSSI to increase the locating accuracy. We use a Commercial Off-the-Shelf (COTS) UHF RFID reader which is integrated with a phased array antenna to evaluate our method. The experiment results from an indoor office environment demonstrate the average distance error of PATL is about 21 cm and the optimized approach achieves an accuracy of 13 cm. This novel 2D localization scheme is a simple, yet promising, solution that is especially applicable to the smart shelf visualized management in storage or retail area.

  4. Architecture and Protocol of a Semantic System Designed for Video Tagging with Sensor Data in Mobile Devices

    PubMed Central

    Macias, Elsa; Lloret, Jaime; Suarez, Alvaro; Garcia, Miguel

    2012-01-01

    Current mobile phones come with several sensors and powerful video cameras. These video cameras can be used to capture good quality scenes, which can be complemented with the information gathered by the sensors also embedded in the phones. For example, the surroundings of a beach recorded by the camera of the mobile phone, jointly with the temperature of the site can let users know via the Internet if the weather is nice enough to swim. In this paper, we present a system that tags the video frames of the video recorded from mobile phones with the data collected by the embedded sensors. The tagged video is uploaded to a video server, which is placed on the Internet and is accessible by any user. The proposed system uses a semantic approach with the stored information in order to make easy and efficient video searches. Our experimental results show that it is possible to tag video frames in real time and send the tagged video to the server with very low packet delay variations. As far as we know there is not any other application developed as the one presented in this paper. PMID:22438753

  5. Architecture and protocol of a semantic system designed for video tagging with sensor data in mobile devices.

    PubMed

    Macias, Elsa; Lloret, Jaime; Suarez, Alvaro; Garcia, Miguel

    2012-01-01

    Current mobile phones come with several sensors and powerful video cameras. These video cameras can be used to capture good quality scenes, which can be complemented with the information gathered by the sensors also embedded in the phones. For example, the surroundings of a beach recorded by the camera of the mobile phone, jointly with the temperature of the site can let users know via the Internet if the weather is nice enough to swim. In this paper, we present a system that tags the video frames of the video recorded from mobile phones with the data collected by the embedded sensors. The tagged video is uploaded to a video server, which is placed on the Internet and is accessible by any user. The proposed system uses a semantic approach with the stored information in order to make easy and efficient video searches. Our experimental results show that it is possible to tag video frames in real time and send the tagged video to the server with very low packet delay variations. As far as we know there is not any other application developed as the one presented in this paper.

  6. Fluorescence lifetime endoscopy using TCSPC for the measurement of FRET in live cells

    PubMed Central

    Fruhwirth, Gilbert O.; Ameer-Beg, Simon; Cook, Richard; Watson, Timothy; Ng, Tony; Festy, Frederic

    2010-01-01

    Development of remote imaging for diagnostic purposes has progressed dramatically since endoscopy began in the 1960’s. The recent advent of a clinically licensed intensity-based fluorescence micro-endoscopic instrument has offered the prospect of real-time cellular resolution imaging. However, interrogating protein-protein interactions deep inside living tissue requires precise fluorescence lifetime measurements to derive the Förster resonance energy transfer between two tagged fluorescent markers. We developed a new instrument combining remote fiber endoscopic cellular-resolution imaging with TCSPC-FLIM technology to interrogate and discriminate mixed fluorochrome labeled beads and expressible GFP/TagRFP tags within live cells. Endoscopic-FLIM (e-FLIM) data was validated by comparison with data acquired via conventional FLIM and e-FLIM was found to be accurate for both bright bead and dim live cell samples. The fiber based micro-endoscope allowed remote imaging of 4 µm and 10 µm beads within a thick Matrigel matrix with confident fluorophore discrimination using lifetime information. More importantly, this new technique enabled us to reliably measure protein-protein interactions in live cells embedded in a 3D matrix, as demonstrated by the dimerization of the fluorescent protein-tagged membrane receptor CXCR4. This cell-based application successfully demonstrated the suitability and great potential of this new technique for in vivo pre-clinical biomedical and possibly human clinical applications. PMID:20588974

  7. Electronic tagging and integrated product intelligence

    NASA Astrophysics Data System (ADS)

    Swerdlow, Martin; Weeks, Brian

    1996-03-01

    The advent of 'intelligent,' electronic data bearing tags is set to revolutionize the way industrial and retail products are identified and tracked throughout their life cycles. The dominant system for unique identification today is the bar code, which is based on printed symbology and regulated by the International Article Numbering Association. Bar codes provide users with significant operational advantages and generate considerable added value to packaging companies, product manufacturers, distributors and retailers, across supply chains in many different sectors, from retailing, to baggage handling and industrial components, e.g., for vehicles or aircraft. Electronic tags offer the potential to: (1) record and store more complex data about the product or any modifications which occur during its life cycle; (2) access (and up-date) stored data in real time in a way which does not involve contact with the product or article; (3) overcome the limitations imposed by systems which rely on line-of-sight access to stored data. Companies are now beginning to consider how electronic data tags can be used, not only to improve the efficiency of their supply chain processes, but also to revolutionize the way they do business. This paper reviews the applications and business opportunities for electronic tags and outlines CEST's strategy for achieving an 'open' standard which will ensure that tags from different vendors can co-exist on an international basis.

  8. Using kittens to unlock photo-sharing website datasets for environmental applications

    NASA Astrophysics Data System (ADS)

    Gascoin, Simon

    2016-04-01

    Mining photo-sharing websites is a promising approach to complement in situ and satellite observations of the environment, however a challenge is to deal with the large degree of noise inherent to online social datasets. Here I explored the value of the Flickr image hosting website database to monitor the snow cover in the Pyrenees. Using the Flickr application programming interface (API) I queried all the public images metadata tagged at least with one of the following words: "snow", "neige", "nieve", "neu" (snow in French, Spanish and Catalan languages). The search was limited to the geo-tagged pictures taken in the Pyrenees area. However, the number of public pictures available in the Flickr database for a given time interval depends on several factors, including the Flickr website popularity and the development of digital photography. Thus, I also searched for all Flickr images tagged with "chat", "gat" or "gato" (cat in French, Spanish and Catalan languages). The tag "cat" was not considered in order to exclude the results from North America where Flickr got popular earlier than in Europe. The number of "cat" images per month was used to fit a model of the number of images uploaded in Flickr with time. This model was used to remove this trend in the numbers of snow-tagged photographs. The resulting time series was compared to a time series of the snow cover area derived from the MODIS satellite over the same region. Both datasets are well correlated; in particular they exhibit the same seasonal evolution, although the inter-annual variabilities are less similar. I will also discuss which other factors may explain the main discrepancies in order to further decrease the noise in the Flickr dataset.

  9. A simple and robust approach to immobilization of antibody fragments.

    PubMed

    Ikonomova, Svetlana P; He, Ziming; Karlsson, Amy J

    2016-08-01

    Antibody fragments, such as the single-chain variable fragment (scFv), have much potential in research and diagnostics because of their antigen-binding ability similar to a full-sized antibody and their ease of production in microorganisms. Some applications of antibody fragments require immobilization on a surface, and we have established a simple immobilization method that is based on the biotin-streptavidin interaction and does not require a separate purification step. We genetically fused two biotinylation tags-the biotin carboxyl carrier protein (BCCP) or the AviTag minimal sequence-to six different scFvs (scFv13R4, scFvD10, scFv26-10, scFv3, scFv5, and scFv12) for site-specific biotinylation in vivo by endogenous biotin ligases produced by Escherichia coli. The biotinylated scFvs were immobilized onto streptavidin-coated plates directly from cell lysates, and immobilization was detected through enzyme-linked immunosorbent assays. All scFvs fusions were successfully immobilized, and scFvs biotinylated via the BCCP tag tended to immobilize better than those biotinylated via the AviTag, even when biotinylation efficiency was improved with the biotin ligase BirA. The ability of immobilized scFvs to bind antigens was confirmed using scFv13R4 and scFvD10 with their respective targets β-galactosidase and bacteriophage lambda head protein D (gpD). The immobilized scFv13R4 bound to β-galactosidase at the same level for both biotinylation tags when the surface was saturated with the scFv, and immobilized scFvs retained their functionality for at least 100days after immobilization. The simplicity and robustness of our method make it a promising approach for future applications that require antibody fragment immobilization. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Application of an E. coli signal sequence as a versatile inclusion body tag.

    PubMed

    Jong, Wouter S P; Vikström, David; Houben, Diane; van den Berg van Saparoea, H Bart; de Gier, Jan-Willem; Luirink, Joen

    2017-03-21

    Heterologous protein production in Escherichia coli often suffers from bottlenecks such as proteolytic degradation, complex purification procedures and toxicity towards the expression host. Production of proteins in an insoluble form in inclusion bodies (IBs) can alleviate these problems. Unfortunately, the propensity of heterologous proteins to form IBs is variable and difficult to predict. Hence, fusing the target protein to an aggregation prone polypeptide or IB-tag is a useful strategy to produce difficult-to-express proteins in an insoluble form. When screening for signal sequences that mediate optimal targeting of heterologous proteins to the periplasmic space of E. coli, we observed that fusion to the 39 amino acid signal sequence of E. coli TorA (ssTorA) did not promote targeting but rather directed high-level expression of the human proteins hEGF, Pla2 and IL-3 in IBs. Further analysis revealed that ssTorA even mediated IB formation of the highly soluble endogenous E. coli proteins TrxA and MBP. The ssTorA also induced aggregation when fused to the C-terminus of target proteins and appeared functional as IB-tag in E. coli K-12 as well as B strains. An additive effect on IB-formation was observed upon fusion of multiple ssTorA sequences in tandem, provoking almost complete aggregation of TrxA and MBP. The ssTorA-moiety was successfully used to produce the intrinsically unstable hEGF and the toxic fusion partner SymE, demonstrating its applicability as an IB-tag for difficult-to-express and toxic proteins. We present proof-of-concept for the use of ssTorA as a small, versatile tag for robust E. coli-based expression of heterologous proteins in IBs.

  11. Identification of a Pair of Phospholipid:Diacylglycerol Acyltransferases from Developing Flax (Linum usitatissimum L.) Seed Catalyzing the Selective Production of Trilinolenin*

    PubMed Central

    Pan, Xue; Siloto, Rodrigo M. P.; Wickramarathna, Aruna D.; Mietkiewska, Elzbieta; Weselake, Randall J.

    2013-01-01

    The oil from flax (Linum usitatissimum L.) has high amounts of α-linolenic acid (ALA; 18:3cisΔ9,12,15) and is one of the richest sources of omega-3 polyunsaturated fatty acids (ω-3-PUFAs). To produce ∼57% ALA in triacylglycerol (TAG), it is likely that flax contains enzymes that can efficiently transfer ALA to TAG. To test this hypothesis, we conducted a systematic characterization of TAG-synthesizing enzymes from flax. We identified several genes encoding acyl-CoA:diacylglycerol acyltransferases (DGATs) and phospholipid:diacylglycerol acyltransferases (PDATs) from the flax genome database. Due to recent genome duplication, duplicated gene pairs have been identified for all genes except DGAT2-2. Analysis of gene expression indicated that two DGAT1, two DGAT2, and four PDAT genes were preferentially expressed in flax embryos. Yeast functional analysis showed that DGAT1, DGAT2, and two PDAT enzymes restored TAG synthesis when produced recombinantly in yeast H1246 strain. The activity of particular PDAT enzymes (LuPDAT1 and LuPDAT2) was stimulated by the presence of ALA. Further seed-specific expression of flax genes in Arabidopsis thaliana indicated that DGAT1, PDAT1, and PDAT2 had significant effects on seed oil phenotype. Overall, this study indicated the existence of unique PDAT enzymes from flax that are able to preferentially catalyze the synthesis of TAG containing ALA acyl moieties. The identified LuPDATs may have practical applications for increasing the accumulation of ALA and other polyunsaturated fatty acids in oilseeds for food and industrial applications. PMID:23824186

  12. Ocean Heat Content Reveals Secrets of Fish Migrations

    PubMed Central

    Luo, Jiangang; Ault, Jerald S.; Shay, Lynn K.; Hoolihan, John P.; Prince, Eric D.; Brown, Craig A.; Rooker, Jay R.

    2015-01-01

    For centuries, the mechanisms surrounding spatially complex animal migrations have intrigued scientists and the public. We present a new methodology using ocean heat content (OHC), a habitat metric that is normally a fundamental part of hurricane intensity forecasting, to estimate movements and migration of satellite-tagged marine fishes. Previous satellite-tagging research of fishes using archival depth, temperature and light data for geolocations have been too coarse to resolve detailed ocean habitat utilization. We combined tag data with OHC estimated from ocean circulation and transport models in an optimization framework that substantially improved geolocation accuracy over SST-based tracks. The OHC-based movement track provided the first quantitative evidence that many of the tagged highly migratory fishes displayed affinities for ocean fronts and eddies. The OHC method provides a new quantitative tool for studying dynamic use of ocean habitats, migration processes and responses to environmental changes by fishes, and further, improves ocean animal tracking and extends satellite-based animal tracking data for other potential physical, ecological, and fisheries applications. PMID:26484541

  13. Computational exposure assessment of electromagnetic fields generated by an RFID system for mother--newborn identity reconfirmation.

    PubMed

    Fiocchi, Serena; Parazzini, Marta; Paglialonga, Alessia; Ravazzani, Paolo

    2011-07-01

    Radio frequency identification (RFID) is an innovative technology currently applied in a large number of industrial and consumer applications. The spread of RFID technology does not correspond to a parallel increase in studies on its possible impact on health in terms of electromagnetic field (EMF) exposure. The aim of this paper is to estimate, by computational techniques, the EMF generated by passive RFID systems for mother-newborn identity reconfirmation. The computation was performed on realistic models of newborn and mother for three different reader positions. The compliance with EMF exposure guidelines was investigated as a function of the change in reader-tag specifications (magnetic field threshold and maximum distance of the reader to awake the tag) and time of use of the reader close to the body. The results show that attention should be paid to the identification of the optimal reader-tag technical specifications to be used in this type of application. That should be done by an accurate exposure assessment investigation, in particular for newborn exposure. The need to reduce the exposure time as much as possible indicates the importance of specific training on the practical applications of the RFID (DATALOGIC J-series, Bologna, Italy) device. Copyright © 2011 Wiley-Liss, Inc.

  14. Reversible Immobilization of Proteins in Sensors and Solid-State Nanopores.

    PubMed

    Ananth, Adithya; Genua, María; Aissaoui, Nesrine; Díaz, Leire; Eisele, Nico B; Frey, Steffen; Dekker, Cees; Richter, Ralf P; Görlich, Dirk

    2018-05-01

    The controlled functionalization of surfaces with proteins is crucial for many analytical methods in life science research and biomedical applications. Here, a coating for silica-based surfaces is established which enables stable and selective immobilization of proteins with controlled orientation and tunable surface density. The coating is reusable, retains functionality upon long-term storage in air, and is applicable to surfaces of complex geometry. The protein anchoring method is validated on planar surfaces, and then a method is developed to measure the anchoring process in real time using silicon nitride solid-state nanopores. For surface attachment, polyhistidine tags that are site specifically introduced into recombinant proteins are exploited, and the yeast nucleoporin Nsp1 is used as model protein. Contrary to the commonly used covalent thiol chemistry, the anchoring of proteins via polyhistidine tag is reversible, permitting to take proteins off and replace them by other ones. Such switching in real time in experiments on individual nanopores is monitored using ion conductivity. Finally, it is demonstrated that silica and gold surfaces can be orthogonally functionalized to accommodate polyhistidine-tagged proteins on silica but prevent protein binding to gold, which extends the applicability of this surface functionalization method to even more complex sensor devices. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Monovalent Strep-Tactin for strong and site-specific tethering in nanospectroscopy.

    PubMed

    Baumann, Fabian; Bauer, Magnus S; Milles, Lukas F; Alexandrovich, Alexander; Gaub, Hermann E; Pippig, Diana A

    2016-01-01

    Strep-Tactin, an engineered form of streptavidin, binds avidly to the genetically encoded peptide Strep-tag II in a manner comparable to streptavidin binding to biotin. These interactions have been used in protein purification and detection applications. However, in single-molecule studies, for example using atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS), the tetravalency of these systems impedes the measurement of monodispersed data. Here, we introduce a monovalent form of Strep-Tactin that harbours a unique binding site for Strep-tag II and a single cysteine that allows Strep-Tactin to specifically attach to the atomic force microscope cantilever and form a consistent pulling geometry to obtain homogeneous rupture data. Using AFM-SMFS, the mechanical properties of the interaction between Strep-tag II and monovalent Strep-Tactin were characterized. Rupture forces comparable to biotin:streptavidin unbinding were observed. Using titin kinase and green fluorescent protein, we show that monovalent Strep-Tactin is generally applicable to protein unfolding experiments. We expect monovalent Strep-Tactin to be a reliable anchoring tool for a range of single-molecule studies.

  16. Monovalent Strep-Tactin for strong and site-specific tethering in nanospectroscopy

    NASA Astrophysics Data System (ADS)

    Baumann, Fabian; Bauer, Magnus S.; Milles, Lukas F.; Alexandrovich, Alexander; Gaub, Hermann E.; Pippig, Diana A.

    2016-01-01

    Strep-Tactin, an engineered form of streptavidin, binds avidly to the genetically encoded peptide Strep-tag II in a manner comparable to streptavidin binding to biotin. These interactions have been used in protein purification and detection applications. However, in single-molecule studies, for example using atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS), the tetravalency of these systems impedes the measurement of monodispersed data. Here, we introduce a monovalent form of Strep-Tactin that harbours a unique binding site for Strep-tag II and a single cysteine that allows Strep-Tactin to specifically attach to the atomic force microscope cantilever and form a consistent pulling geometry to obtain homogeneous rupture data. Using AFM-SMFS, the mechanical properties of the interaction between Strep-tag II and monovalent Strep-Tactin were characterized. Rupture forces comparable to biotin:streptavidin unbinding were observed. Using titin kinase and green fluorescent protein, we show that monovalent Strep-Tactin is generally applicable to protein unfolding experiments. We expect monovalent Strep-Tactin to be a reliable anchoring tool for a range of single-molecule studies.

  17. Enzyme-linked electrochemical DNA ligation assay using magnetic beads.

    PubMed

    Stejskalová, Eva; Horáková, Petra; Vacek, Jan; Bowater, Richard P; Fojta, Miroslav

    2014-07-01

    DNA ligases are essential enzymes in all cells and have been proposed as targets for novel antibiotics. Efficient DNA ligase activity assays are thus required for applications in biomedical research. Here we present an enzyme-linked electrochemical assay based on two terminally tagged probes forming a nicked junction upon hybridization with a template DNA. Nicked DNA bearing a 5' biotin tag is immobilized on the surface of streptavidin-coated magnetic beads, and ligated product is detected via a 3' digoxigenin tag recognized by monoclonal antibody-alkaline phosphatase conjugate. Enzymatic conversion of napht-1-yl phosphate to napht-1-ol enables sensitive detection of the voltammetric signal on a pyrolytic graphite electrode. The technique was tested under optimal conditions and various situations limiting or precluding the ligation reaction (such as DNA substrates lacking 5'-phosphate or containing a base mismatch at the nick junction, or application of incompatible cofactor), and utilized for the analysis of the nick-joining activity of a range of recombinant Escherichia coli DNA ligase constructs. The novel technique provides a fast, versatile, specific, and sensitive electrochemical assay of DNA ligase activity.

  18. Exploring the Host Parasitism of the Migratory Plant-Parasitic Nematode Ditylenchus destuctor by Expressed Sequence Tags Analysis

    PubMed Central

    Peng, Huan; Gao, Bing-li; Kong, Ling-an; Yu, Qing; Huang, Wen-kun; He, Xu-feng; Long, Hai-bo; Peng, De-liang

    2013-01-01

    The potato rot nematode, Ditylenchus destructor, is a very destructive nematode pest on many agriculturally important crops worldwide, but the molecular characterization of its parasitism of plant has been limited. The effectors involved in nematode parasitism of plant for several sedentary endo-parasitic nematodes such as Heterodera glycines, Globodera rostochiensis and Meloidogyne incognita have been identified and extensively studied over the past two decades. Ditylenchus destructor, as a migratory plant parasitic nematode, has different feeding behavior, life cycle and host response. Comparing the transcriptome and parasitome among different types of plant-parasitic nematodes is the way to understand more fully the parasitic mechanism of plant nematodes. We undertook the approach of sequencing expressed sequence tags (ESTs) derived from a mixed stage cDNA library of D. destructor. This is the first study of D. destructor ESTs. A total of 9800 ESTs were grouped into 5008 clusters including 3606 singletons and 1402 multi-member contigs, representing a catalog of D. destructor genes. Implementing a bioinformatics' workflow, we found 1391 clusters have no match in the available gene database; 31 clusters only have similarities to genes identified from D. africanus, the most closely related species to D. destructor; 1991 clusters were annotated using Gene Ontology (GO); 1550 clusters were assigned enzyme commission (EC) numbers; and 1211 clusters were mapped to 181 KEGG biochemical pathways. 22 ESTs had similarities to reported nematode effectors. Interestedly, most of the effectors identified in this study are involved in host cell wall degradation or modification, such as 1,4-beta-glucanse, 1,3-beta-glucanse, pectate lyase, chitinases and expansin, or host defense suppression such as calreticulin, annexin and venom allergen-like protein. This result implies that the migratory plant-parasitic nematode D. destructor secrets similar effectors to those of sedentary plant nematodes. Finally we further characterized the two D. destructor expansin proteins. PMID:23922743

  19. Exploring the Presence of microDNAs in Prostate Cancer Cell Lines, Tissue, and Sera of Prostate Cancer Patients and its Possible Application as Biomarker

    DTIC Science & Technology

    2015-08-01

    Sequence tags were mapped on the human reference genome using the Novoalign software. Only those tags... the linear islands to create a novel junctional sequence that does not exist in the genome . Thus the PE- sequence of a fragment that breaks at or...identified in cancer cell lines. (b) Median percent GC content of microDNAs and the genomic sequences up- or downstream of the source loci are

  20. The Extent of DoD Influence on the Development and Application of Radio Frequency (RFID) Technology in the Civilian Sector

    DTIC Science & Technology

    2005-06-01

    the consumer. When you buy a Rolex watch for 20 3 American Industry was not particularly interested in tagging animals. Rather, tagging...you accept some risk that the watch you are buying is not actually a Rolex . The problem comes when you buy an item from a reputable store who...MOTHER OF INVENTION 1. RADAR is Born September 1, 1939. The world watches in horror as Germany invades Poland. World War II has officially begun

  1. Feasibility of Radio Frequency Identification (RFID) and Item Unique Identification (IUID) in the Marine Corps Small Arms Weapons Tracking System

    DTIC Science & Technology

    2008-12-04

    tracked. The RFID tag can be of various designs, materials , and/or sizes and hold a variable amount of information. Each tag is composed of three...efficiency in acquiring material from suppliers and in delivering to units in the field (Business Wire, 2005). The military learned an important lesson...RFID Supply-chain applications of RFID are beneficial to both the DoD and suppliers. RFID technology has enabled the recording of material transfer

  2. 50 CFR 648.7 - Recordkeeping and reporting requirements.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... amount by species (by market category, if applicable); price per unit by species (by market category, if applicable) or total value by species (by market category, if applicable); port landed; cage tag numbers for..., butterfish, scup, or black sea bass at sea must submit information identical to that required by paragraph (a...

  3. Research, development and application of noncombustible Beta fiber structures. [for Apollo

    NASA Technical Reports Server (NTRS)

    Dillon, J. J.; Cobb, E. S.

    1975-01-01

    Beta fiber was selected as the primary material for flexible fibrous structures used in spacecraft and crew systems applications in the Apollo program because it was noncombustible in a 100 percent oxygen atmosphere up to 16.5 psia. It met NASA criteria for outgassing, toxicity, odor, and crew comfort, and possessed sufficient durability to last through the mission. Topics discussed include: study of spacecraft applications; design of Beta fiber textile structures to meet the requirements; selection of surface treatments (finishes, coatings, and printing systems) to impart the required durability and special functional use to the textile structures; development of sewing and fabrication techniques; and testing and evaluation programs, and development of production sources.

  4. TRANSMIT evaluation : transit application baseline data report

    DOT National Transportation Integrated Search

    2002-03-01

    As part of the FY99 earmarked ITS Integration Program, the TRANSMIT program received funds to expand the use of the E-ZPass toll tag readers to other transportation applications in New Jersey. Operated by Transcom for agencies in NY, NJ, and CT, TRAN...

  5. NuLat: 3D Event Reconstruction of a ROL Detector for Neutrino Detection and Background Rejection

    NASA Astrophysics Data System (ADS)

    Yokley, Zachary; NuLat Collaboration

    2015-04-01

    NuLat is a proposed very-short baseline reactor antineutrino experiment that employs a unique detector design, a Ragahavan Optical Lattice (ROL), developed for the LENS solar neutrino experiment. The 3D lattice provides high spatial and temporal resolution and allows for energy deposition in each voxel to be determined independently of other voxels, as well as the time sequence associated with each voxel energy deposition. This unique feature arises from two independent means to spatially locate energy deposits: via timing and via optical channeling. NuLat, the first application of a ROL detector targeting physics results, will measure the reactor antineutrino flux at very short baselines via inverse beta decay (IBD). The ROL design of NuLat makes possible the reconstruction of positron energy with little contamination due to the annihilation gammas which smear the positron energy resolution in a traditional detector. IBD events are cleanly tagged via temporal and spatial coincidence of neutron capture in the vertex voxel or nearest neighbors. This talk will present work on IBD event reconstruction in NuLat and its likely impact on sterile neutrino detection via operation in higher background locations enabled by its superior rejection of backgrounds. This research has been funded in part by the National Science Foundation on Award Numbers 1001394 and 1001078.

  6. A Liquid Phase Affinity Capture Assay Using Magnetic Beads to Study Protein-Protein Interaction: The Poliovirus-Nanobody Example

    PubMed Central

    Schotte, Lise; Rombaut, Bart; Thys, Bert

    2012-01-01

    In this article, a simple, quantitative, liquid phase affinity capture assay is presented. Provided that one protein can be tagged and another protein labeled, this method can be implemented for the investigation of protein-protein interactions. It is based on one hand on the recognition of the tagged protein by cobalt coated magnetic beads and on the other hand on the interaction between the tagged protein and a second specific protein that is labeled. First, the labeled and tagged proteins are mixed and incubated at room temperature. The magnetic beads, that recognize the tag, are added and the bound fraction of labeled protein is separated from the unbound fraction using magnets. The amount of labeled protein that is captured can be determined in an indirect way by measuring the signal of the labeled protein remained in the unbound fraction. The described liquid phase affinity assay is extremely useful when conformational conversion sensitive proteins are assayed. The development and application of the assay is demonstrated for the interaction between poliovirus and poliovirus recognizing nanobodies1. Since poliovirus is sensitive to conformational conversion2 when attached to a solid surface (unpublished results), the use of ELISA is limited and a liquid phase based system should therefore be preferred. An example of a liquid phase based system often used in polioresearch3,4 is the micro protein A-immunoprecipitation test5. Even though this test has proven its applicability, it requires an Fc-structure, which is absent in the nanobodies6,7. However, as another opportunity, these interesting and stable single-domain antibodies8 can be easily engineered with different tags. The widely used (His)6-tag shows affinity for bivalent ions such as nickel or cobalt, which can on their turn be easily coated on magnetic beads. We therefore developed this simple quantitative affinity capture assay based on cobalt coated magnetic beads. Poliovirus was labeled with 35S to enable unhindered interaction with the nanobodies and to make a quantitative detection feasible. The method is easy to perform and can be established with a low cost, which is further supported by the possibility of effectively regenerating the magnetic beads. PMID:22688388

  7. Comparison of the structures of triacylglycerols from native and transgenic medium-chain fatty acid-enriched rape seed oil by liquid chromatography--atmospheric pressure chemical ionization ion-trap mass spectrometry (LC-APCI-ITMS).

    PubMed

    Beermann, Christopher; Winterling, Nadine; Green, Angelika; Möbius, Michael; Schmitt, Joachim J; Boehm, Günther

    2007-04-01

    The sn position of fatty acids in seed oil lipids affects physiological function in pharmaceutical and dietary applications. In this study the composition of acyl-chain substituents in the sn positions of glycerol backbones in triacylglycerols (TAG) have been compared. TAG from native and transgenic medium-chain fatty acid-enriched rape seed oil were analyzed by reversed-phase high performance liquid chromatography coupled with online atmospheric-pressure chemical ionization ion-trap mass spectrometry. The transformation of summer rape with thioesterase and 3-ketoacyl-[ACP]-synthase genes of Cuphea lanceolata led to increased expression of 1.5% (w/w) caprylic acid (8:0), 6.7% (w/w) capric acid (10:0), 0.9% (w/w) lauric acid (12:0), and 0.2% (w/w) myristic acid (14:0). In contrast, linoleic (18:2n6) and alpha-linolenic acid (18:3n3) levels decreased compared with the original seed oil. The TAG sn position distribution of fatty acids was also modified. The original oil included eleven unique TAG species whereas the transgenic oil contained sixty. Twenty species were common to both oils. The transgenic oil included trioctadecenoyl-glycerol (18:1/18:1/18:1) and trioctadecatrienoyl-glycerol (18:3/18:3/18:3) whereas the native oil included only the latter. The transgenic TAG were dominated by combinations of caprylic, capric, lauric, myrisitic, palmitic (16:0), stearic (18:0), oleic (18:1n9), linoleic, arachidic (20:0), behenic (22:0), and lignoceric acids (24:0), which accounted for 52% of the total fat. In the original TAG palmitic, stearic, oleic, and linoleic acids accounted for 50% of the total fat. Medium-chain triacylglycerols with capric and lauric acids combined with stearic, oleic, linoleic, alpha-linolenic, arachidic, and gondoic acids (20:1n9) accounted for 25% of the transgenic oil. The medium-chain fatty acids were mainly integrated into the sn-1/3 position combined with the essential linoleic and alpha-linolenic acids at the sn-2 position. Eight species contained caprylic, capric, and lauric acids in the sn-2 position. The appearance of new TAG in the transgenic oil illustrates the extensive effect of genetic modification on fat metabolism by transformed plants and offers interesting possibilities for improved enteral applications.

  8. Reverse Engineering and Security Evaluation of Commercial Tags for RFID-Based IoT Applications.

    PubMed

    Fernández-Caramés, Tiago M; Fraga-Lamas, Paula; Suárez-Albela, Manuel; Castedo, Luis

    2016-12-24

    The Internet of Things (IoT) is a distributed system of physical objects that requires the seamless integration of hardware (e.g., sensors, actuators, electronics) and network communications in order to collect and exchange data. IoT smart objects need to be somehow identified to determine the origin of the data and to automatically detect the elements around us. One of the best positioned technologies to perform identification is RFID (Radio Frequency Identification), which in the last years has gained a lot of popularity in applications like access control, payment cards or logistics. Despite its popularity, RFID security has not been properly handled in numerous applications. To foster security in such applications, this article includes three main contributions. First, in order to establish the basics, a detailed review of the most common flaws found in RFID-based IoT systems is provided, including the latest attacks described in the literature. Second, a novel methodology that eases the detection and mitigation of such flaws is presented. Third, the latest RFID security tools are analyzed and the methodology proposed is applied through one of them (Proxmark 3) to validate it. Thus, the methodology is tested in different scenarios where tags are commonly used for identification. In such systems it was possible to clone transponders, extract information, and even emulate both tags and readers. Therefore, it is shown that the methodology proposed is useful for auditing security and reverse engineering RFID communications in IoT applications. It must be noted that, although this paper is aimed at fostering RFID communications security in IoT applications, the methodology can be applied to any RFID communications protocol.

  9. Reverse Engineering and Security Evaluation of Commercial Tags for RFID-Based IoT Applications

    PubMed Central

    Fernández-Caramés, Tiago M.; Fraga-Lamas, Paula; Suárez-Albela, Manuel; Castedo, Luis

    2016-01-01

    The Internet of Things (IoT) is a distributed system of physical objects that requires the seamless integration of hardware (e.g., sensors, actuators, electronics) and network communications in order to collect and exchange data. IoT smart objects need to be somehow identified to determine the origin of the data and to automatically detect the elements around us. One of the best positioned technologies to perform identification is RFID (Radio Frequency Identification), which in the last years has gained a lot of popularity in applications like access control, payment cards or logistics. Despite its popularity, RFID security has not been properly handled in numerous applications. To foster security in such applications, this article includes three main contributions. First, in order to establish the basics, a detailed review of the most common flaws found in RFID-based IoT systems is provided, including the latest attacks described in the literature. Second, a novel methodology that eases the detection and mitigation of such flaws is presented. Third, the latest RFID security tools are analyzed and the methodology proposed is applied through one of them (Proxmark 3) to validate it. Thus, the methodology is tested in different scenarios where tags are commonly used for identification. In such systems it was possible to clone transponders, extract information, and even emulate both tags and readers. Therefore, it is shown that the methodology proposed is useful for auditing security and reverse engineering RFID communications in IoT applications. It must be noted that, although this paper is aimed at fostering RFID communications security in IoT applications, the methodology can be applied to any RFID communications protocol. PMID:28029119

  10. Enzyme resistant carbohydrate based micro-scale materials from sugar beet (Beta vulgaris L.) pulp for food and pharmaceutical applications

    USDA-ARS?s Scientific Manuscript database

    Bio-based micro scale materials are increasingly used in functional food and pharmaceutical applications. The present study produced carbohydrate-based micro scale tubular materials from sugar beet (Beta vulgaris L.) pulp (SBP), a by-product of sugar beet processing. The isolated carbohydrates wer...

  11. Purification and characterization of a GH43 β-xylosidase from Enterobacter sp. identified and cloned from forest soil bacteria.

    PubMed

    Campos, Eleonora; Negro Alvarez, María José; Sabarís di Lorenzo, Gonzalo; Gonzalez, Sergio; Rorig, Marcela; Talia, Paola; Grasso, Daniel H; Sáez, Felicia; Manzanares Secades, Paloma; Ballesteros Perdices, Mercedes; Cataldi, Angel A

    2014-01-01

    The use of lignocellulosic biomass for second generation biofuels requires optimization of enzymatic breakdown of plant cell walls. In this work, cellulolytic bacteria were isolated from a native and two cultivated forest soil samples. Amplification of glycosyl hydrolases was attempted by using a low stringency-degenerate primer PCR strategy, using total soil DNA and bulk DNA pooled from positive colonies as template. A set of primers was designed based on Acidothermus cellulolyticus genome, by search of conserved domains of glycosyl hydrolases (GH) families of interest. Using this approach, a fragment containing an open reading frame (ORF) with 98% identity to a putative GH43 beta-xylosidase coding gene from Enterobacter cloacae was amplified and cloned. The full protein was expressed in Escherichia coli as N-terminal or C-terminal His-tagged fusions and purified under native conditions. Only N-terminal fusion protein, His-Xyl43, presented beta-xylosidase activity. On pNPX, optimal activity was achieved at pH 6 and 40 °C and Km and Kcat values were 2.92 mM and 1.32 seg(-1), respectively. Activity was also demonstrated on xylobiose (X2), with Km 17.8 mM and Kcat 380 s(-1). These results demonstrated that Xyl43 is a functional beta-xylosidase and it is the first evidence of this activity for Enterobacter sp. Copyright © 2013 Elsevier GmbH. All rights reserved.

  12. Micro-power dissipation device described

    NASA Astrophysics Data System (ADS)

    Mao, X.; Zhou, L.; Zhou, J.

    1985-11-01

    The common-emitter current gain beta of a common two-pole transistor is generally below 250. They are referred to as high-beta or high gain transistors when the beta of such transistors exceeds 300. When the beta of a transistor is higher than 1,000, it is called a super-beta transistor (SBT) or supergain transistor. The micropower dissipation type has the widest applications among the high-beta. Micropower dissipation high-beta means that there is a high gain or a superhigh gain under a microcurrent. The device is widely used in small signal-detection systems and stereo audio equipment because of their characteristics of high gain, low frequency and low noise under small signals.

  13. Superglue from bacteria: unbreakable bridges for protein nanotechnology.

    PubMed

    Veggiani, Gianluca; Zakeri, Bijan; Howarth, Mark

    2014-10-01

    Biotechnology is often limited by weak interactions. We suggest that an ideal interaction between proteins would be covalent, specific, require addition of only a peptide tag to the protein of interest, and form under a wide range of conditions. Here we summarize peptide tags that are able to form spontaneous amide bonds, based on harnessing reactions of adhesion proteins from the bacterium Streptococcus pyogenes. These include the irreversible peptide-protein interaction of SpyTag with SpyCatcher, as well as irreversible peptide-peptide interactions via SpyLigase. We describe existing applications, including polymerization to enhance cancer cell capture, assembly of living biomaterial, access to diverse protein shapes, and improved enzyme resilience. We also indicate future opportunities for resisting biological force and extending the scope of protein nanotechnology. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Performance of low-power RFID tags based on modulated backscattering

    NASA Astrophysics Data System (ADS)

    Mhanna, Zeinab; Sibille, Alain; Contreras, Richard

    2017-02-01

    Ultra Wideband (UWB) modulated backscattering (MBS) passive Radio-Frequency IDentification (RFID) systems provide a promising solution to overcome many limitations of current narrowband RFID devices. This work addresses the performance of such systems from the point of view of the radio channel between the readers and the tags. Such systems will likely combine several readers, in order to provide both the detection and localization of tags operating in MBS. Two successive measurements campaigns have been carried out in an indoor reference scenario environment. The first is intended to verify the methods and serves as a way to validate the RFID backscattering measurement setup. The second represents a real use case for RFID application and allows one to quantitatively analyze the path loss of the backscattering propagation channel. xml:lang="fr"

  15. Development of novel wireless sensor for food quality detection

    NASA Astrophysics Data System (ADS)

    Son Nguyen, Dat; Ngan Le, Nguyen; Phat Lam, Tan; Fribourg-Blanc, Eric; Chien Dang, Mau; Tedjini, Smail

    2015-12-01

    In this paper we present a wireless sensor for the monitoring of food quality. We integrate sensing capability into ultrahigh frequency (UHF) radio-frequency identification (RFID) tags through the relationship between the physical read-range and permittivity of the object we label with the RFID tags. Using the known variations of food permittivity as a function of time, we can detect the contamination time at which a food product becomes unacceptable for consumption based on the measurement of read-range with the as-designed sensing tags. This low-cost UHF RFID passive sensor was designed and experimentally tested on beef, pork, and cheese with the same storage conditions as in supermarkets. The agreement between the experimental and simulation results show the potential of this technique for practical application in food-quality tracking.

  16. Perilipin 1 Mediates Lipid Metabolism Homeostasis and Inhibits Inflammatory Cytokine Synthesis in Bovine Adipocytes

    PubMed Central

    Zhang, Shiqi; Liu, Guowen; Xu, Chuang; Liu, Lei; Zhang, Qiang; Xu, Qiushi; Jia, Hongdou; Li, Xiaobing; Li, Xinwei

    2018-01-01

    Dairy cows with ketosis displayed lipid metabolic disorder and high inflammatory levels. Adipose tissue is an active lipid metabolism and endocrine tissue and is closely related to lipid metabolism homeostasis and inflammation. Perilipin 1 (PLIN1), an adipocyte-specific lipid-coated protein, may be involved in the above physiological function. The aim of this study is to investigate the role of PLIN1 in lipid metabolism regulation and inflammatory factor synthesis in cow adipocytes. The results showed that PLIN1 overexpression upregulated the expression of fatty acid and triglyceride (TAG) synthesis molecule sterol regulator element-binding protein-1c (SREBP-1c) and its target genes, diacylglycerol acyltransferase (DGAT) 1, and DGAT2, but inhibited the expression of lipolysis enzymes hormone-sensitive lipase (HSL) and CGI-58 for adipose triglyceride lipase (ATGL), thus augmenting the fatty acids and TAG synthesis and inhibiting lipolysis. Importantly, PLIN1 overexpression inhibited the activation of the NF-κB inflammatory pathway and decreased the expression and content of tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6) induced by lipopolysaccharide. Conversely, PLIN1 silencing inhibited TAG synthesis, promoted lipolysis, and overinduced the activation of the NF-κB inflammatory pathway in cow adipocytes. In ketotic cows, the expression of PLIN1 was markedly decreased, whereas lipid mobilization, NF-κB pathway, and downstream inflammatory cytokines were overinduced in adipose tissue. Taken together, these results indicate that PLIN1 can maintain lipid metabolism homeostasis and inhibit the NF-κB inflammatory pathway in adipocytes. However, low levels of PLIN1 reduced the inhibitory effect on fat mobilization, NF-κB pathway, and inflammatory cytokine synthesis in ketotic cows. PMID:29593725

  17. Perilipin 1 Mediates Lipid Metabolism Homeostasis and Inhibits Inflammatory Cytokine Synthesis in Bovine Adipocytes.

    PubMed

    Zhang, Shiqi; Liu, Guowen; Xu, Chuang; Liu, Lei; Zhang, Qiang; Xu, Qiushi; Jia, Hongdou; Li, Xiaobing; Li, Xinwei

    2018-01-01

    Dairy cows with ketosis displayed lipid metabolic disorder and high inflammatory levels. Adipose tissue is an active lipid metabolism and endocrine tissue and is closely related to lipid metabolism homeostasis and inflammation. Perilipin 1 (PLIN1), an adipocyte-specific lipid-coated protein, may be involved in the above physiological function. The aim of this study is to investigate the role of PLIN1 in lipid metabolism regulation and inflammatory factor synthesis in cow adipocytes. The results showed that PLIN1 overexpression upregulated the expression of fatty acid and triglyceride (TAG) synthesis molecule sterol regulator element-binding protein-1c (SREBP-1c) and its target genes, diacylglycerol acyltransferase (DGAT) 1, and DGAT2, but inhibited the expression of lipolysis enzymes hormone-sensitive lipase (HSL) and CGI-58 for adipose triglyceride lipase (ATGL), thus augmenting the fatty acids and TAG synthesis and inhibiting lipolysis. Importantly, PLIN1 overexpression inhibited the activation of the NF-κB inflammatory pathway and decreased the expression and content of tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6) induced by lipopolysaccharide. Conversely, PLIN1 silencing inhibited TAG synthesis, promoted lipolysis, and overinduced the activation of the NF-κB inflammatory pathway in cow adipocytes. In ketotic cows, the expression of PLIN1 was markedly decreased, whereas lipid mobilization, NF-κB pathway, and downstream inflammatory cytokines were overinduced in adipose tissue. Taken together, these results indicate that PLIN1 can maintain lipid metabolism homeostasis and inhibit the NF-κB inflammatory pathway in adipocytes. However, low levels of PLIN1 reduced the inhibitory effect on fat mobilization, NF-κB pathway, and inflammatory cytokine synthesis in ketotic cows.

  18. Raman Imaging in Cell Membranes, Lipid-Rich Organelles, and Lipid Bilayers.

    PubMed

    Syed, Aleem; Smith, Emily A

    2017-06-12

    Raman-based optical imaging is a promising analytical tool for noninvasive, label-free chemical imaging of lipid bilayers and cellular membranes. Imaging using spontaneous Raman scattering suffers from a low intensity that hinders its use in some cellular applications. However, developments in coherent Raman imaging, surface-enhanced Raman imaging, and tip-enhanced Raman imaging have enabled video-rate imaging, excellent detection limits, and nanometer spatial resolution, respectively. After a brief introduction to these commonly used Raman imaging techniques for cell membrane studies, this review discusses selected applications of these modalities for chemical imaging of membrane proteins and lipids. Finally, recent developments in chemical tags for Raman imaging and their applications in the analysis of selected cell membrane components are summarized. Ongoing developments toward improving the temporal and spatial resolution of Raman imaging and small-molecule tags with strong Raman scattering cross sections continue to expand the utility of Raman imaging for diverse cell membrane studies.

  19. Modifying the ECC-based grouping-proof RFID system to increase inpatient medication safety.

    PubMed

    Ko, Wen-Tsai; Chiou, Shin-Yan; Lu, Erl-Huei; Chang, Henry Ker-Chang

    2014-09-01

    RFID technology is increasingly used in applications that require tracking, identification, and authentication. It attaches RFID-readable tags to objects for identification and execution of specific RFID-enabled applications. Recently, research has focused on the use of grouping-proofs for preserving privacy in RFID applications, wherein a proof of two or more tags must be simultaneously scanned. In 2010, a privacy-preserving grouping proof protocol for RFID based on ECC in public-key cryptosystem was proposed but was shown to be vulnerable to tracking attacks. A proposed enhancement protocol was also shown to have defects which prevented proper execution. In 2012, Lin et al. proposed a more efficient RFID ECC-based grouping proof protocol to promote inpatient medication safety. However, we found this protocol is also vulnerable to tracking and impersonation attacks. We then propose a secure privacy-preserving RFID grouping proof protocol for inpatient medication safety and demonstrate its resistance to such attacks.

  20. Nanoparticle-mediated local and remote manipulation of protein aggregation.

    PubMed

    Kogan, Marcelo J; Bastus, Neus G; Amigo, Roger; Grillo-Bosch, Dolors; Araya, Eyleen; Turiel, Antonio; Labarta, Amilcar; Giralt, Ernest; Puntes, Victor F

    2006-01-01

    The local heat delivered by metallic nanoparticles selectively attached to their target can be used as a molecular surgery to safely remove toxic and clogging aggregates. We apply this principle to protein aggregates, in particular to the amyloid beta protein (Abeta) involved in Alzheimer's disease (AD), a neurodegenerative disease where unnaturally folded Abeta proteins self-assemble and deposit forming amyloid fibrils and plaques. We show the possibility to remotely redissolve these deposits and to interfere with their growth, using the local heat dissipated by gold nanoparticles (AuNP) selectively attached to the aggregates and irradiated with low gigahertz electromagnetic fields. Simultaneous tagging and manipulation by AuNP of Abeta at different stages of aggregation allow both, noninvasive exploration and dissolution of molecular aggregates.

  1. Insight into the coordination and the binding sites of Cu(2+) by the histidyl-6-tag using experimental and computational tools.

    PubMed

    Watly, Joanna; Simonovsky, Eyal; Wieczorek, Robert; Barbosa, Nuno; Miller, Yifat; Kozlowski, Henryk

    2014-07-07

    His-tags are specific sequences containing six to nine subsequent histydyl residues, and they are used for purification of recombinant proteins by use of IMAC chromatography. Such polyhistydyl tags, often used in molecular biology, can be also found in nature. Proteins containing histidine-rich domains play a critical role in many life functions in both prokaryote and eukaryote organisms. Binding mode and the thermodynamic properties of the system depend on the specific metal ion and the histidine sequence. Despite the wide application of the His-tag for purification of proteins, little is known about the properties of metal-binding to such tag domains. This inspired us to undertake detailed studies on the coordination of Cu(2+) ion to hexa-His-tag. Experiments were performed using the potentiometric, UV-visible, CD, and EPR techniques. In addition, molecular dynamics (MD) simulations and density functional theory (DFT) calculations were applied. The experimental studies have shown that the Cu(2+) ion binds most likely to two imidazoles and one, two, or three amide nitrogens, depending on the pH. The structures and stabilities of the complexes for the Cu(2+)-Ac-(His)6-NH2 system using experimental and computational tools were established. Polymorphic binding states are suggested, with a possibility of the formation of α-helix structure induced by metal ion coordination. Metal ion is bound to various pairs of imidazole moieties derived from the tag with different efficiencies. The coordination sphere around the metal ion is completed by molecules of water. Finally, the Cu(2+) binding by Ac-(His)6-NH2 is much more efficient compared to other multihistidine protein domains.

  2. Electronic individual identification of zebrafish using radio frequency identification (RFID) microtags.

    PubMed

    Cousin, Xavier; Daouk, Tarek; Péan, Samuel; Lyphout, Laura; Schwartz, Marie-Elise; Bégout, Marie-Laure

    2012-08-15

    Although individual electronic tagging using passive integrated acoustic (PIT) tags is established, it is mainly for fish >60 mm in length and is unsuitable for fish of <30 mm, like zebrafish. We used radio frequency identification (RFID) microtags (1 mm in diameter and 6 mm in length, with a mass of ~10 mg) to individually identify juvenile zebrafish (length 16-42 mm, mass 138-776 mg) for the first time, and studied the effects of intracoelomic implantation on fish survival and microtag loss, growth, spawning and exploratory behaviour. After 5.5 months, both high survival (82%) and low microtag loss (11%) were achieved. The smallest surviving fish weighed 178 mg, and success in microtag reading was 73% for the size class 350-450 mg (26 mm). Greater success was achieved when fish were larger at the time of tagging but no negative effects on growth were observed for any size class and some tagged fish spawned. No significant differences in behavioural responses could be detected between tagged fish and untagged controls after 2 months. Overall, the results suggest that the tagging method is highly suitable for fish as small as zebrafish juveniles. We think this method will provide significant advances for researchers of the ever-growing fish model community and more generally for all small-fish users. Tagging is essential when one needs to identify fish (e.g. particular genotypes with no external cue), to run longitudinal monitoring of individual biological traits (e.g. growth) or to repeat assays with the same individual at discrete points in time (e.g. behaviour studies). Such a method will find applications in physiology, genetics, behaviour and (eco)toxicology fields.

  3. A novel surgical marking system for small peripheral lung nodules based on radio frequency identification technology: Feasibility study in a canine model.

    PubMed

    Kojima, Fumitsugu; Sato, Toshihiko; Takahata, Hiromi; Okada, Minoru; Sugiura, Tadao; Oshiro, Osamu; Date, Hiroshi; Nakamura, Tatsuo

    2014-04-01

    We investigated the feasibility and accuracy of a novel surgical marking system based on radiofrequency identification (RFID) technology for the localization of small peripheral lung nodules (SPLNs) in a canine model. The system consists of 4 components: (1) micro RFID tags (13.56 MHz, 1.0 × 1.0 × 0.8 mm), (2) a tag delivery system with a bronchoscope, (3) a wand-shaped locating probe (10-mm diameter), and (4) a signal processing unit with audio interface. Before the operation, pseudolesions mimicking SPLNs were prepared in 7 dogs by injecting colored collagen. By use of a computed tomographic (CT) guide, an RFID tag was placed via a bronchoscope close to each target lesion. This was then followed by scanning with the locating probe, and wedge resection was performed when possible. Operators can locate the tag by following the sound emitted by the system, which exhibits tone changes according to the tag-probe distance. The primary outcome measure was the rate of wedge resection with good margins. A total of 10 pseudolesions imitating SPLNs were selected as targets. During thoracoscopic procedures, 9 of 10 tags were detected by the system within a median of 27 seconds. Wedge resections were performed for these 9 lesions with a median margin of 11 mm. The single failure was caused by tag dislocation to the central airway. Successful localization and wedge resection of pseudolesions with appropriate margins were accomplished in an experimental setting. Our RFID marking system has future applications for accurately locating SPLNs in a clinical setting. Copyright © 2014 The American Association for Thoracic Surgery. Published by Mosby, Inc. All rights reserved.

  4. Contrasting Life Strategies of Viruses that Infect Photo- and Heterotrophic Bacteria, as Revealed by Viral Tagging

    PubMed Central

    Deng, Li; Gregory, Ann; Yilmaz, Suzan; Poulos, Bonnie T.; Hugenholtz, Philip; Sullivan, Matthew B.

    2012-01-01

    ABSTRACT Ocean viruses are ubiquitous and abundant and play important roles in global biogeochemical cycles by means of their mortality, horizontal gene transfer, and manipulation of host metabolism. However, the obstacles involved in linking viruses to their hosts in a high-throughput manner bottlenecks our ability to understand virus-host interactions in complex communities. We have developed a method called viral tagging (VT), which combines mixtures of host cells and fluorescent viruses with flow cytometry. We investigated multiple viruses which infect each of two model marine bacteria that represent the slow-growing, photoautotrophic genus Synechococcus (Cyanobacteria) and the fast-growing, heterotrophic genus Pseudoalteromonas (Gammaproteobacteria). Overall, viral tagging results for viral infection were consistent with plaque and liquid infection assays for cyanobacterial myo-, podo- and siphoviruses and some (myo- and podoviruses) but not all (four siphoviruses) heterotrophic bacterial viruses. Virus-tagged Pseudoalteromonas organisms were proportional to the added viruses under varied infection conditions (virus-bacterium ratios), while no more than 50% of the Synechococcus organisms were virus tagged even at viral abundances that exceeded (5 to 10×) that of their hosts. Further, we found that host growth phase minimally impacts the fraction of virus-tagged Synechococcus organisms while greatly affecting phage adsorption to Pseudoalteromonas. Together these findings suggest that at least two contrasting viral life strategies exist in the oceans and that they likely reflect adaptation to their host microbes. Looking forward to the point at which the virus-tagging signature is well understood (e.g., for Synechococcus), application to natural communities should begin to provide population genomic data at the proper scale for predictively modeling two of the most abundant biological entities on Earth. PMID:23111870

  5. a Chiral Tagging Strategy for Determining Absolute Configuration and Enantiomeric Excess by Molecular Rotational Spectroscopy

    NASA Astrophysics Data System (ADS)

    Evangelisti, Luca; Caminati, Walther; Patterson, David; Thomas, Javix; Xu, Yunjie; West, Channing; Pate, Brooks

    2017-06-01

    The introduction of three wave mixing rotational spectroscopy by Patterson, Schnell, and Doyle [1,2] has expanded applications of molecular rotational spectroscopy into the field of chiral analysis. Chiral analysis of a molecule is the quantitative measurement of the relative abundances of all stereoisomers of the molecule and these include both diastereomers (with distinct molecular rotational spectra) and enantiomers (with equivalent molecular rotational spectra). This work adapts a common strategy in chiral analysis of enantiomers to molecular rotational spectroscopy. A "chiral tag" is attached to the molecule of interest by making a weakly bound complex in a pulsed jet expansion. When this tag molecule is enantiopure, it will create diastereomeric complexes with the two enantiomers of the molecule being analyzed and these can be differentiated by molecule rotational spectroscopy. Identifying the structure of this complex, with knowledge of the absolute configuration of the tag, establishes the absolute configuration of the molecule of interest. Furthermore, the diastereomer complex spectra can be used to determine the enantiomeric excess of the sample. The ability to perform chiral analysis will be illustrated by a study of solketal using propylene oxide as the tag. The possibility of using current methods of quantum chemistry to assign a specific structure to the chiral tag complex will be discussed. Finally, chiral tag rotational spectroscopy offers a "gold standard" method for determining the absolute configuration of the molecule through determination of the substitution structure of the complex. When this measurement is possible, rotational spectroscopy can deliver a quantitative three dimensional structure of the molecule with correct stereochemistry as the analysis output. [1] David Patterson, Melanie Schnell, John M. Doyle, Nature 497, 475 (2013). [2] David Patterson, John M. Doyle, Phys. Rev. Lett. 111, 023008 (2013).

  6. Direct Detection of Biotinylated Proteins by Mass Spectrometry

    PubMed Central

    2015-01-01

    Mass spectrometric strategies to identify protein subpopulations involved in specific biological functions rely on covalently tagging biotin to proteins using various chemical modification methods. The biotin tag is primarily used for enrichment of the targeted subpopulation for subsequent mass spectrometry (MS) analysis. A limitation of these strategies is that MS analysis does not easily discriminate unlabeled contaminants from the labeled protein subpopulation under study. To solve this problem, we developed a flexible method that only relies on direct MS detection of biotin-tagged proteins called “Direct Detection of Biotin-containing Tags” (DiDBiT). Compared with conventional targeted proteomic strategies, DiDBiT improves direct detection of biotinylated proteins ∼200 fold. We show that DiDBiT is applicable to several protein labeling protocols in cell culture and in vivo using cell permeable NHS-biotin and incorporation of the noncanonical amino acid, azidohomoalanine (AHA), into newly synthesized proteins, followed by click chemistry tagging with biotin. We demonstrate that DiDBiT improves the direct detection of biotin-tagged newly synthesized peptides more than 20-fold compared to conventional methods. With the increased sensitivity afforded by DiDBiT, we demonstrate the MS detection of newly synthesized proteins labeled in vivo in the rodent nervous system with unprecedented temporal resolution as short as 3 h. PMID:25117199

  7. Experimental studies of high-accuracy RFID localization with channel impairments

    NASA Astrophysics Data System (ADS)

    Pauls, Eric; Zhang, Yimin D.

    2015-05-01

    Radio frequency identification (RFID) systems present an incredibly cost-effective and easy-to-implement solution to close-range localization. One of the important applications of a passive RFID system is to determine the reader position through multilateration based on the estimated distances between the reader and multiple distributed reference tags obtained from, e.g., the received signal strength indicator (RSSI) readings. In practice, the achievable accuracy of passive RFID reader localization suffers from many factors, such as the distorted RSSI reading due to channel impairments in terms of the susceptibility to reader antenna patterns and multipath propagation. Previous studies have shown that the accuracy of passive RFID localization can be significantly improved by properly modeling and compensating for such channel impairments. The objective of this paper is to report experimental study results that validate the effectiveness of such approaches for high-accuracy RFID localization. We also examine a number of practical issues arising in the underlying problem that limit the accuracy of reader-tag distance measurements and, therefore, the estimated reader localization. These issues include the variations in tag radiation characteristics for similar tags, effects of tag orientations, and reader RSS quantization and measurement errors. As such, this paper reveals valuable insights of the issues and solutions toward achieving high-accuracy passive RFID localization.

  8. Accelerometer-derived activity correlates with volitional swimming speed in lake sturgeon (Acipenser fulvescens)

    USGS Publications Warehouse

    Thiem, J.D.; Dawson, J.W.; Gleiss, A.C.; Martins, E.G.; Haro, Alexander J.; Castro-Santos, Theodore R.; Danylchuk, A.J.; Wilson, R.P.; Cooke, S.J.

    2015-01-01

    Quantifying fine-scale locomotor behaviours associated with different activities is challenging for free-swimming fish.Biologging and biotelemetry tools can help address this problem. An open channel flume was used to generate volitionalswimming speed (Us) estimates of cultured lake sturgeon (Acipenser fulvescens Rafinesque, 1817) and these were paired withsimultaneously recorded accelerometer-derived metrics of activity obtained from three types of data-storage tags. This studyexamined whether a predictive relationship could be established between four different activity metrics (tail-beat frequency(TBF), tail-beat acceleration amplitude (TBAA), overall dynamic body acceleration (ODBA), and vectorial dynamic body acceleration(VeDBA)) and the swimming speed of A. fulvescens. Volitional Us of sturgeon ranged from 0.48 to 2.70 m·s−1 (0.51–3.18 bodylengths (BL) · s−1). Swimming speed increased linearly with all accelerometer-derived metrics, and when all tag types werecombined, Us increased 0.46 BL·s−1 for every 1 Hz increase in TBF, and 0.94, 0.61, and 0.94 BL·s−1 for every 1g increase in TBAA,ODBA, and VeDBA, respectively. Predictive relationships varied among tag types and tag-specific parameter estimates of Us arepresented for all metrics. This use of acceleration data-storage tags demonstrated their applicability for the field quantificationof sturgeon swimming speed.

  9. The Monitoring and Affinity Purification of Proteins Using Dual Tags with Tetracysteine Motifs

    NASA Astrophysics Data System (ADS)

    Giannone, Richard J.; Liu, Yie; Wang, Yisong

    Identification and characterization of protein-protein interaction networks is essential for the elucidation of biochemical mechanisms and cellular function. Affinity purification in combination with liquid chromatography-tandem mass spectrometry (LC-MS/MS) has emerged as a very powerful tactic for the identification of specific protein-protein interactions. In this chapter, we describe a comprehensive methodology that uses our recently developed dual-tag affinity purification system for the enrichment and identification of mammalian protein complexes. The protocol covers a series of separate but sequentially related techniques focused on the facile monitoring and purification of a dual-tagged protein of interest and its interacting partners via a system built with tetracysteine motifs and various combinations of affinity tags. Using human telomeric repeat binding factor 2 (TRF2) as an example, we demonstrate the power of the system in terms of bait protein recovery after dual-tag affinity purification, detection of bait protein subcellular localization and expression, and successful identification of known and potentially novel TRF2 interacting proteins. Although the protocol described here has been optimized for the identification and characterization of TRF2-associated proteins, it is, in principle, applicable to the study of any other mammalian protein complexes that may be of interest to the research community.

  10. Self-etching adhesive on intact enamel, with and without pre-etching.

    PubMed

    Devarasa, G M; Subba Reddy, V V; Chaitra, N L; Swarna, Y M

    2012-05-01

    Bond strengths of composite resin to enamel using self-etch adhesive (SEA) Clearfil SE bond system on intact enamel and enamel pre-etched with phosphoric acid were compared. The objective was to determine if the pre-etching would increase the bond strengths of the SEA systems to intact enamel and to evaluate the effect of pre-etching on bond formation of self-etch adhesives on intact enamel. Labial surfaces of 40 caries free permanent upper central and lateral incisors were cleaned, sectioned of their roots. All specimens were mounted on acrylic block and divided randomly into four groups. In two groups the application of self-etch adhesive, Clearfil SE bond was carried as per manufacturer's instructions, composite cylinders were built, whereas in the other two groups, 37% phosphoric acid etching was done before the application of self-etching adhesives. Then the resin tags were analyzed using scanning electron microscope and shear bond strength was measured using Instron universal testing machine. When phosphoric acid was used, there was significant increase in the depth of penetration of resin tags and in the Shear Bond Strength of composite to enamel. The results indicate that out of both treatment groups, pre-etching the intact enamel with 37% phosphoric acid resulted in formation of longer resin tags and higher depth of penetration of resin tags of the Clearfil SE bond, and attaining higher bond strength of the Clearfil SE bond to intact enamel. Copyright © 2011 Wiley Periodicals, Inc.

  11. A Three-Parameter Generalisation of the Beta-Binomial Distribution with Applications

    DTIC Science & Technology

    1987-07-01

    York. Rust, R.T. and Klompmaker, J.E. (1981). Improving the estimation procedure for the beta binomial t.v. exposure model. Journal of Marketing ... Research . 18, 442-448. Sabavala, D.J. and Morrison, D.G. (1977). Television show loyalty: a beta- binomial model using recall data. Journal of Advertiuing

  12. The Role of CREB in CML

    DTIC Science & Technology

    2008-02-01

    Feng YQ et al. Anti-beta s- ribozyme reduces beta s mRNA levels in transgenic mice: Potential application to the gene therapy of sickle cell anemia... ribozymes . RNA 2003;9:1254–1263. 13 Pace BS, Qian X, Ofori-Acquah SF. Selective inhibition of beta-globin RNA transcripts by antisense RNA molecules. Cell

  13. Positronium signature in organic liquid scintillators for neutrino experiments

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Franco, D.; Consolati, G.; Trezzi, D.

    2011-01-15

    Electron antineutrinos are commonly detected in liquid scintillator experiments via inverse {beta} decay by looking at the coincidence between the reaction products: neutrons and positrons. Prior to positron annihilation, an electron-positron pair may form an orthopositronium (o-Ps) state, with a mean lifetime of a few nanoseconds. Even if the o-Ps decay is speeded up by spin-flip or pick-off effects, it may introduce distortions in the photon emission time distribution, crucial for position reconstruction and pulse shape discrimination algorithms in antineutrino experiments. Reversing the problem, the o-Ps-induced time distortion represents a new signature for tagging antineutrinos in liquid scintillator. In thismore » article, we report the results of measurements of the o-Ps formation probability and lifetime for the most used solvents for organic liquid scintillators in neutrino physics (pseudocumene, linear alkyl benzene, phenylxylylethane, and dodecane). We characterize also a mixture of pseudocumene +1.5 g/l of 2,5-diphenyloxazole, a fluor acting as wavelength shifter. In the second part of the article, we demonstrate that the o-Ps-induced distortion of the scintillation photon emission time distributions represent an optimal signature for tagging positrons on an event by event basis, potentially enhancing the antineutrino detection.« less

  14. Studies of a biochemical factory: tomato trichome deep expressed sequence tag sequencing and proteomics.

    PubMed

    Schilmiller, Anthony L; Miner, Dennis P; Larson, Matthew; McDowell, Eric; Gang, David R; Wilkerson, Curtis; Last, Robert L

    2010-07-01

    Shotgun proteomics analysis allows hundreds of proteins to be identified and quantified from a single sample at relatively low cost. Extensive DNA sequence information is a prerequisite for shotgun proteomics, and it is ideal to have sequence for the organism being studied rather than from related species or accessions. While this requirement has limited the set of organisms that are candidates for this approach, next generation sequencing technologies make it feasible to obtain deep DNA sequence coverage from any organism. As part of our studies of specialized (secondary) metabolism in tomato (Solanum lycopersicum) trichomes, 454 sequencing of cDNA was combined with shotgun proteomics analyses to obtain in-depth profiles of genes and proteins expressed in leaf and stem glandular trichomes of 3-week-old plants. The expressed sequence tag and proteomics data sets combined with metabolite analysis led to the discovery and characterization of a sesquiterpene synthase that produces beta-caryophyllene and alpha-humulene from E,E-farnesyl diphosphate in trichomes of leaf but not of stem. This analysis demonstrates the utility of combining high-throughput cDNA sequencing with proteomics experiments in a target tissue. These data can be used for dissection of other biochemical processes in these specialized epidermal cells.

  15. Bifidobacterium breve with α-linolenic acid alters the composition, distribution and transcription factor activity associated with metabolism and absorption of fat

    PubMed Central

    Patterson, Elaine; Wall, Rebecca; Lisai, Sara; Ross, R. Paul; Dinan, Timothy G.; Cryan, John F.; Fitzgerald, Gerald F.; Banni, Sebastiano; Quigley, Eamonn M.; Shanahan, Fergus; Stanton, Catherine

    2017-01-01

    This study focused on the mechanisms that fatty acid conjugating strains - Bifidobacterium breve NCIMB 702258 and Bifidobacterium breve DPC 6330 - influence lipid metabolism when ingested with α-linolenic acid (ALA) enriched diet. Four groups of BALB/c mice received ALA enriched diet (3% (w/w)) either alone or in combination with B. breve NCIMB 702258 or B. breve DPC 6330 (109 CFU/day) or unsupplemented control diet for six weeks. The overall n-3 PUFA score was increased in all groups receiving the ALA enriched diet. Hepatic peroxisomal beta oxidation increased following supplementation of the ALA enriched diet with B. breve (P < 0.05) and so the ability of the strains to produce c9t11 conjugated linoleic acid (CLA) was identified in adipose tissue. Furthermore, a strain specific effect of B. breve NCIMB 702258 was found on the endocannabinoid system (ECS). Liver triglycerides (TAG) were reduced following ALA supplementation, compared with unsupplemented controls (P < 0.01) while intervention with B. breve further reduced liver TAG (P < 0.01), compared with the ALA enriched control. These data indicate that the interactions of the gut microbiota with fatty acid metabolism directly affect host health by modulating n-3 PUFA score and the ECS. PMID:28265110

  16. Bifidobacterium breve with α-linolenic acid alters the composition, distribution and transcription factor activity associated with metabolism and absorption of fat.

    PubMed

    Patterson, Elaine; Wall, Rebecca; Lisai, Sara; Ross, R Paul; Dinan, Timothy G; Cryan, John F; Fitzgerald, Gerald F; Banni, Sebastiano; Quigley, Eamonn M; Shanahan, Fergus; Stanton, Catherine

    2017-03-07

    This study focused on the mechanisms that fatty acid conjugating strains - Bifidobacterium breve NCIMB 702258 and Bifidobacterium breve DPC 6330 - influence lipid metabolism when ingested with α-linolenic acid (ALA) enriched diet. Four groups of BALB/c mice received ALA enriched diet (3% (w/w)) either alone or in combination with B. breve NCIMB 702258 or B. breve DPC 6330 (10 9 CFU/day) or unsupplemented control diet for six weeks. The overall n-3 PUFA score was increased in all groups receiving the ALA enriched diet. Hepatic peroxisomal beta oxidation increased following supplementation of the ALA enriched diet with B. breve (P < 0.05) and so the ability of the strains to produce c9t11 conjugated linoleic acid (CLA) was identified in adipose tissue. Furthermore, a strain specific effect of B. breve NCIMB 702258 was found on the endocannabinoid system (ECS). Liver triglycerides (TAG) were reduced following ALA supplementation, compared with unsupplemented controls (P < 0.01) while intervention with B. breve further reduced liver TAG (P < 0.01), compared with the ALA enriched control. These data indicate that the interactions of the gut microbiota with fatty acid metabolism directly affect host health by modulating n-3 PUFA score and the ECS.

  17. The Low Energy Neutrino Spectrometry (LENS) Experiment and LENS prototype, μLENS, initial results

    NASA Astrophysics Data System (ADS)

    Yokley, Zachary

    2012-03-01

    LENS is a low energy solar neutrino detector that will measure the solar neutrino spectrum above 115 keV, >95% of the solar neutrino flux, in real time. The fundamental neutrino reaction in LENS is charged-current based capture on 115-In detected in a liquid scintillator medium. The reaction yields the prompt emission of an electron and the delayed emission of 2 gamma rays that serve as a time & space coincidence tag. Sufficient spatial resolution is used to exploit this signature and suppress background, particularly due to 115-In beta decay. A novel design of optical segmentation (Scintillation Lattice or SL) channels the signal light along the three primary axes. The channeling is achieved via total internal reflection by suitable low index gaps in the segmentation. The spatial resolution of a nuclear event is obtained digitally, much more precisely than possible by common time of flight methods. Advanced Geant4 analysis methods have been developed to suppress adequately the severe background due to 115-In beta decay, achieving at the same time high detection efficiency. LENS physics and detection methods along with initial results characterizing light transport in the as built μLENS prototype will be presented.

  18. Development and application of nonflammable, high-temperature beta fibers

    NASA Technical Reports Server (NTRS)

    Dawn, Frederic S.

    1989-01-01

    Recent advances in fiber technology have contributed to the success of the U.S. space program. The inorganic fiber Beta, developed as a result of efforts begun in the early 1960's and heightened following the January 27, 1967 Apollo fire is unique among inorganic and organic fibers. It has been developed into woven, nonwoven, knitted, braided, coated and printed structures. All of these were used extensively for the Apollo, Skylab, Apollo-Soyuz test project, space shuttle, Spacelab, and satellite programs. In addition to being used successfully in the space program, Beta fibers are being used commercially as firesafe fabrics in homes, hospitals, institutions, public buildings, aircraft, and public transportation, wherever total nonflammability is required. One of the most unique applications of the Beta composite structure is the roofing material for the 80,000-seat Detroit Lion's Silverdome and 5 square miles of the Jeddah International Airport in Saudi Arabia. This fiber has been successfully incorporated into 165 major public construction projects around the globe. The United States alone has used more than 12 million square yards of the material. Beta fiber has been used successfully to date and has a promising future with unlimited potential for both space and commercial application. Efforts are currently underway to improve Beta fiber to meet the requirements of extended service life for the Space Station Freedom, lunar outpost, and Mars exploration missions.

  19. Successful application of preimplantation genetic diagnosis for beta-thalassaemia and sickle cell anaemia in Italy.

    PubMed

    Chamayou, S; Alecci, C; Ragolia, C; Giambona, A; Siciliano, S; Maggio, A; Fichera, M; Guglielmino, A

    2002-05-01

    In Italy, the autosomal recessive diseases beta-thalassaemia and sickle cell anaemia are so widespread that in some regions they can be defined as 'social diseases'. In this study, nine clinical applications of preimplantation genetic diagnosis (PGD) were performed for beta-thalassaemia and sickle cell anaemia on seven Sicilian couples and carriers of beta-globin gene mutations. The studied mutations were: Cd39, HbS, IVS1 nt1, IVS1 nt6 and IVS1 nt110. ICSI was performed with partner's sperm on 131 out of 147 retrieved oocytes, and this resulted in 72 zygotes; 32 embryos were successfully biopsied on day 3. The biopsied blastomeres were lysed and the beta-globin alleles amplified by nested PCR. The mutation diagnosis was performed by restriction enzyme digestion and reverse dot-blot. The amplification efficacy was 97.2%. The genotype study of non-transferred and surplus embryos showed that the allele drop-out rate was 8.6%. Seventeen embryos were transferred in utero on day 4. All couples received an embryo transfer; of the four pregnancies obtained, three resulted in live births and one miscarried at 11 weeks. Prenatal diagnosis at the 11th week and miscarriage material analysis confirmed the PGD results. These studies represent the first successful application of PGD for beta-thalassaemia and sickle cell anaemia in Italy.

  20. RAMTaB: Robust Alignment of Multi-Tag Bioimages

    PubMed Central

    Raza, Shan-e-Ahmed; Humayun, Ahmad; Abouna, Sylvie; Nattkemper, Tim W.; Epstein, David B. A.; Khan, Michael; Rajpoot, Nasir M.

    2012-01-01

    Background In recent years, new microscopic imaging techniques have evolved to allow us to visualize several different proteins (or other biomolecules) in a visual field. Analysis of protein co-localization becomes viable because molecules can interact only when they are located close to each other. We present a novel approach to align images in a multi-tag fluorescence image stack. The proposed approach is applicable to multi-tag bioimaging systems which (a) acquire fluorescence images by sequential staining and (b) simultaneously capture a phase contrast image corresponding to each of the fluorescence images. To the best of our knowledge, there is no existing method in the literature, which addresses simultaneous registration of multi-tag bioimages and selection of the reference image in order to maximize the overall overlap between the images. Methodology/Principal Findings We employ a block-based method for registration, which yields a confidence measure to indicate the accuracy of our registration results. We derive a shift metric in order to select the Reference Image with Maximal Overlap (RIMO), in turn minimizing the total amount of non-overlapping signal for a given number of tags. Experimental results show that the Robust Alignment of Multi-Tag Bioimages (RAMTaB) framework is robust to variations in contrast and illumination, yields sub-pixel accuracy, and successfully selects the reference image resulting in maximum overlap. The registration results are also shown to significantly improve any follow-up protein co-localization studies. Conclusions For the discovery of protein complexes and of functional protein networks within a cell, alignment of the tag images in a multi-tag fluorescence image stack is a key pre-processing step. The proposed framework is shown to produce accurate alignment results on both real and synthetic data. Our future work will use the aligned multi-channel fluorescence image data for normal and diseased tissue specimens to analyze molecular co-expression patterns and functional protein networks. PMID:22363510

  1. Design, Engineering and Application of an Amyloidogenic Protein, SBAFP-m1, for use in Nanotechnological Applications

    NASA Astrophysics Data System (ADS)

    Peralta, Maria del Refugio

    Nanotechnology relies on collaborations across scientific disciplines such as physics, engineering, chemistry and biology. In nanotechnology, researchers manipulate molecules on the nanometer scale for various applications, ranging from tissue engineering, nanowire synthesis, and alternative energy devices. By utilizing various biological scaffolds, namely amyloid fibrils, the work of nanometer molecular control can be achieved through the use of self-assembly systems. Here, a systematic design scheme was developed to engineer protein based amyloid fibrils and was successfully applied to the design of two, unique self-assembled monomers, SBAFP-m1 and RGAFP-m1, from naturally occurring ice binding proteins found in insects and plants. A highly idealized, in-register dimer interface was designed and experimentally synthesized and demonstrated to form micron long amyloid fibrils (Chapter 2). The strength and resistance of the designer amyloid fibrils formed by SBAFP-m1 were probed in Chapter 3. Most notably, the ultimate tensile strength of SBAFP-m1 fibrils was experimentally determined to be 2.1 +/- 1.7 GPa, on par with that of naturally occurring amyloid fibrils in literature and steel. The fibrils were found to maintain their beta-sheet structure over a wide range of temperatures, from - 80 °C to 90 °C. Fibrils were resistant to common protein denaturants like 8M urea, 2.5 M guanidine hydrochloride, 2.5 M NaCl, organic solvents (methanol, ethanol, isopropanol and acetone), and across the pH range two to 11. SBAFP-m1 was mutated to add a 5x cysteine tag to the N-terminus, allowing for gold nanoparticle conjugation along the fibril axis (Chapter 4). The gold-conjugated fibrils were then enhanced with silver to produce nanowires. Various attempts to selectively synthesize heterogeneous fibrils from SBAFP-m1 mutants were attempted in Chapter 5. An attempt to de-stabilize the homogeneous fibril assembly through unfavorable homogeneous protein interactions was not successful as homogeneous fibrils continued to form. Engineering an intermolecular disulfide bond across the dimer interface was also unsuccessful due to incomplete product and mixed disulfide formation. Heterogeneous fibril assembly via a SpyTag and SpyCatcher moiety resulted in improper folding of the parent, amyloid proteins, although with further optimization, heterogeneous fibril assembly should be attainable. This work illustrates from start to finish the ability to engineer an extremely strong and highly resistant amyloidogenic protein and its ability to be functionalized as a biological scaffold for the synthesis of nanowires. The potential for the engineered protein, SBAFP-m1, to be used in a variety of other nanotechnological applications is promising.

  2. Total absorption studies of high priority decays for reactor applications: 86 Br and 91 Rb

    DOE PAGES

    Algora, A.; Rice, S.; Guadilla, V.; ...

    2017-09-13

    Preliminary results from beta decay studies of nuclei that are important for reactor applications are presented. The beta decays have been studied using the total absorption technique (TAS) and the pure beams provided by the JYFLTRAP system at the IGISOL facility of the University of Jyväskylä.

  3. Formation of active inclusion bodies induced by hydrophobic self-assembling peptide GFIL8.

    PubMed

    Wang, Xu; Zhou, Bihong; Hu, Weike; Zhao, Qing; Lin, Zhanglin

    2015-06-16

    In the last few decades, several groups have observed that proteins expressed as inclusion bodies (IBs) in bacteria could still be biologically active when terminally fused to an appropriate aggregation-prone partner such as pyruvate oxidase from Paenibacillus polymyxa (PoxB). More recently, we have demonstrated that three amphipathic self-assembling peptides, an alpha helical peptide 18A, a beta-strand peptide ELK16, and a surfactant-like peptide L6KD, have properties that induce target proteins into active IBs. We have developed an efficient protein expression and purification approach for these active IBs by introducing a self-cleavable intein molecule. In this study, the self-assembling peptide GFIL8 (GFILGFIL) with only hydrophobic residues was analyzed, and this peptide effectively induced the formation of cytoplasmic IBs in Escherichia coli when terminally attached to lipase A and amadoriase II. The protein aggregates in cells were confirmed by transmission electron microscopy analysis and retained ~50% of their specific activities relative to the native counterparts. We constructed an expression and separation coupled tag (ESCT) by incorporating an intein molecule, the Mxe GyrA intein. Soluble target proteins were successfully released from active IBs upon cleavage of the intein between the GFIL8 tag and the target protein, which was mediated by dithiothreitol. A variant of GFIL8, GFIL16 (GFILGFILGFILGFIL), improved the ESCT scheme by efficiently eliminating interference from the soluble intein-GFIL8 molecule. The yields of target proteins at the laboratory scale were 3.0-7.5 μg/mg wet cell pellet, which is comparable to the yields from similar ESCT constructs using 18A, ELK16, or the elastin-like peptide tag scheme. The all-hydrophobic self-assembling peptide GFIL8 induced the formation of active IBs in E. coli when terminally attached to target proteins. GFIL8 and its variant GFIL16 can act as a "pull-down" tag to produce purified soluble proteins with reasonable quantity and purity from active aggregates. Owing to the structural simplicity, strong hydrophobicity, and high aggregating efficiency, these peptides can be further explored for enzyme production and immobilization.

  4. 49 CFR 569.5 - Applicability.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ..., DEPARTMENT OF TRANSPORTATION REGROOVED TIRES § 569.5 Applicability. (a) General. Except as provided in paragraph (b) of this section, this part applies to all motor vehicle regrooved or regroovable tires... regrooved or regroovable tires intended solely for export and so labeled or tagged. [34 FR 1150, Jan. 24...

  5. beta-Phenylethylamine modulates acetylcholine release in the rat striatum: involvement of a dopamine D(2) receptor mechanism.

    PubMed

    Kato, M; Ishida, K; Chuma, T; Abe, K; Shigenaga, T; Taguchi, K; Miyatake, T

    2001-04-20

    We examined the effects of beta-phenylethylamine on striatal acetylcholine release in freely moving rats using in vivo microdialysis. beta-Phenylethylamine at 12.5 mg/kg, i.p. did not affect acetylcholine release in the striatum, whereas 25 and 50 mg/kg, i.p. immediately induced an increase in acetylcholine release in the striatum at 15-45 min. This increase following intraperitoneal administration of beta-phenylethylamine (25 mg/kg) was not affected by locally applied SCH-23390 (R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine, 10 microM), a dopamine D(1) receptor antagonist, nor by raclopride (10 microM), a dopamine D(2) receptor antagonist. The increased release of acetylcholine induced by beta-phenylethylamine was suppressed by local infusion of tetrodotoxin (1 microM). In contrast, the extracellular acetylcholine level in the striatum was significantly decreased by local application of beta-phenylethylamine (10 and 100 microM) in the striatum via a microdialysis probe. The decrease was completely blocked by local co-application of raclopride (10 microM). The beta-phenylethylamine-induced decrease in striatal acetylcholine release was not affected by co-perfusion with SCH-23390 (10 microM). These results indicate that systemic administration of beta-phenylethylamine increases acetylcholine release, whereas locally applied beta-phenylethylamine decreases striatal acetylcholine release in freely moving rats. Furthermore, the dopaminergic system, through the dopamine D(2) receptor, is involved in the locally applied beta-phenylethylamine-induced decrease in acetylcholine in the striatum.

  6. Beta-Testing Agreement | FNLCR Staging

    Cancer.gov

    Beta-Testing Agreements are appropriate forlimited term evaluation and applications development of new software, technology, or equipment platforms by the Frederick National Labin collaboration with an external commercial partner. It may

  7. Preparation of novel beta-cyclodextrin functionalized monolith and its application in chiral separation.

    PubMed

    Lv, Yongqin; Mei, Danping; Pan, Xinxin; Tan, Tianwei

    2010-09-15

    A novel beta-cyclodextrin (beta-CD) functionalized organic polymer monolith was prepared by covalently bonding ethylenediamine-beta-CD (EDA-beta-CD) to poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) (poly(GMA-co-EGDMA)) monolith via ring opening reaction of epoxy groups. SEM characterization was performed to confirm the homogeneity of the monolithic polymer. The resulting monolith was then characterized by DSC and XPS elemental analysis to study the thermal stability of the monolith, and to prove the successful immobilization of beta-CD on the polymer substrate. The beta-CD ligand density of 0.68 mmol g(-1) was obtained for the modified monolith, indicating the high reactivity and efficiency of the EDA-beta-CD modifier. The ethylenediamine-beta-CD functionalized monoliths were used for the chiral separation of ibuprofen racemic mixture and showed promising results. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  8. A method to analyze molecular tagging velocimetry data using the Hough transform.

    PubMed

    Sanchez-Gonzalez, R; McManamen, B; Bowersox, R D W; North, S W

    2015-10-01

    The development of a method to analyze molecular tagging velocimetry data based on the Hough transform is presented. This method, based on line fitting, parameterizes the grid lines "written" into a flowfield. Initial proof-of-principle illustration of this method was performed to obtain two-component velocity measurements in the wake of a cylinder in a Mach 4.6 flow, using a data set derived from computational fluid dynamics simulations. The Hough transform is attractive for molecular tagging velocimetry applications since it is capable of discriminating spurious features that can have a biasing effect in the fitting process. Assessment of the precision and accuracy of the method were also performed to show the dependence on analysis window size and signal-to-noise levels. The accuracy of this Hough transform-based method to quantify intersection displacements was determined to be comparable to cross-correlation methods. The employed line parameterization avoids the assumption of linearity in the vicinity of each intersection, which is important in the limit of drastic grid deformations resulting from large velocity gradients common in high-speed flow applications. This Hough transform method has the potential to enable the direct and spatially accurate measurement of local vorticity, which is important in applications involving turbulent flowfields. Finally, two-component velocity determinations using the Hough transform from experimentally obtained images are presented, demonstrating the feasibility of the proposed analysis method.

  9. A 2.4-GHz ISM RF and UWB hybrid RFID real-time locating system for industrial enterprise Internet of Things

    NASA Astrophysics Data System (ADS)

    Zhai, Chuanying; Zou, Zhuo; Zhou, Qin; Mao, Jia; Chen, Qiang; Tenhunen, Hannu; Zheng, Lirong; Xu, Lida

    2017-07-01

    This paper presents a 2.4-GHz radio frequency (RF) and ultra-wide bandwidth (UWB) hybrid real-time locating system (RTLS) for industrial enterprise Internet of Things (IoT). It employs asymmetric wireless link, that is, UWB radio is utilised for accurate positioning up to 10 cm in critical sites, whereas 2.4-GHz RF is used for tag control and coarse positioning in non-critical sites. The specified communication protocol and the adaptive tag synchronisation rate ensure reliable and deterministic access with a scalable system capacity and avoid unpredictable latency and additional energy consumption of retransmissions due to collisions. The tag, consisting of a commercial 2.4-GHz transceiver and a customised application-specific integrated circuit (ASIC) UWB transmitter (Tx), is able to achieve up to 3 years' battery life at 1600 tags per position update second with 1000 mAh battery in one cluster. The time difference of arrival (TDoA)-based positioning experiment at UWB radio is performed on the designed software-defined radio (SDR) platform.

  10. Mapping transcription factor interactome networks using HaloTag protein arrays.

    PubMed

    Yazaki, Junshi; Galli, Mary; Kim, Alice Y; Nito, Kazumasa; Aleman, Fernando; Chang, Katherine N; Carvunis, Anne-Ruxandra; Quan, Rosa; Nguyen, Hien; Song, Liang; Alvarez, José M; Huang, Shao-Shan Carol; Chen, Huaming; Ramachandran, Niroshan; Altmann, Stefan; Gutiérrez, Rodrigo A; Hill, David E; Schroeder, Julian I; Chory, Joanne; LaBaer, Joshua; Vidal, Marc; Braun, Pascal; Ecker, Joseph R

    2016-07-19

    Protein microarrays enable investigation of diverse biochemical properties for thousands of proteins in a single experiment, an unparalleled capacity. Using a high-density system called HaloTag nucleic acid programmable protein array (HaloTag-NAPPA), we created high-density protein arrays comprising 12,000 Arabidopsis ORFs. We used these arrays to query protein-protein interactions for a set of 38 transcription factors and transcriptional regulators (TFs) that function in diverse plant hormone regulatory pathways. The resulting transcription factor interactome network, TF-NAPPA, contains thousands of novel interactions. Validation in a benchmarked in vitro pull-down assay revealed that a random subset of TF-NAPPA validated at the same rate of 64% as a positive reference set of literature-curated interactions. Moreover, using a bimolecular fluorescence complementation (BiFC) assay, we confirmed in planta several interactions of biological interest and determined the interaction localizations for seven pairs. The application of HaloTag-NAPPA technology to plant hormone signaling pathways allowed the identification of many novel transcription factor-protein interactions and led to the development of a proteome-wide plant hormone TF interactome network.

  11. Synthesis of CdTe quantum dot-conjugated CC49 and their application for in vitro imaging of gastric adenocarcinoma cells

    NASA Astrophysics Data System (ADS)

    Zhang, Yun-Peng; Sun, Peng; Zhang, Xu-Rui; Yang, Wu-Li; Si, Cheng-Shuai

    2013-06-01

    The purpose of this experiment was to investigate the visible imaging of gastric adenocarcinoma cells in vitro by targeting tumor-associated glycoprotein 72 (TAG-72) with near-infrared quantum dots (QDs). QDs with an emission wavelength of about 550 to 780 nm were conjugated to CC49 monoclonal antibodies against TAG-72, resulting in a probe named as CC49-QDs. A gastric adenocarcinoma cell line (MGC80-3) expressing high levels of TAG-72 was cultured for fluorescence imaging, and a gastric epithelial cell line (GES-1) was used for the negative control group. Transmission electron microscopy indicated that the average diameter of CC49-QDs was 0.2 nm higher compared with that of the primary QDs. Also, fluorescence spectrum analysis indicated that the CC49-QDs did not have different optical properties compared to the primary QDs. Immunohistochemical examination and in vitro fluorescence imaging of the tumors showed that the CC49-QDs probe could bind TAG-72 expressed on MGC80-3 cells.

  12. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nekoogar, F; Dowla, F

    An IAEA Technical Meeting on Techniques for IAEA Verification of Enrichment Activities identified 'smart tags' as a technology that should be assessed for tracking and locating UF6 cylinders. Although there is vast commercial industry working on RFID systems, the vulnerabilities of commercial products are only beginning to emerge. Most of the commercially off-the-shelf (COTS) RFID systems operate in very narrow frequency bands, making them vulnerable to detection, jamming and tampering and also presenting difficulties when used around metals (i.e. UF6 cylinders). Commercial passive RFID tags have short range, while active RFID tags that provide long ranges have limited lifetimes. Theremore » are also some concerns with the introduction of strong (narrowband) radio frequency signals around radioactive and nuclear materials. Considering the shortcomings of commercial RFID systems, in their current form, they do not offer a promising solution for continuous monitoring and tracking of UF6 cylinders. In this paper, we identify the key challenges faced by commercial RFID systems for monitoring UF6 cylinders, and introduce an ultra-wideband approach for tag/reader communications that addresses most of the identified challenges for IAEA safeguards applications.« less

  13. A capture-recapture survival analysis model for radio-tagged animals

    USGS Publications Warehouse

    Pollock, K.H.; Bunck, C.M.; Winterstein, S.R.; Chen, C.-L.; North, P.M.; Nichols, J.D.

    1995-01-01

    In recent years, survival analysis of radio-tagged animals has developed using methods based on the Kaplan-Meier method used in medical and engineering applications (Pollock et al., 1989a,b). An important assumption of this approach is that all tagged animals with a functioning radio can be relocated at each sampling time with probability 1. This assumption may not always be reasonable in practice. In this paper, we show how a general capture-recapture model can be derived which allows for some probability (less than one) for animals to be relocated. This model is not simply a Jolly-Seber model because it is possible to relocate both dead and live animals, unlike when traditional tagging is used. The model can also be viewed as a generalization of the Kaplan-Meier procedure, thus linking the Jolly-Seber and Kaplan-Meier approaches to survival estimation. We present maximum likelihood estimators and discuss testing between submodels. We also discuss model assumptions and their validity in practice. An example is presented based on canvasback data collected by G. M. Haramis of Patuxent Wildlife Research Center, Laurel, Maryland, USA.

  14. The development of an intermediate-duration tag to characterize the diving behavior of large whales.

    PubMed

    Mate, Bruce R; Irvine, Ladd M; Palacios, Daniel M

    2017-01-01

    The development of high-resolution archival tag technologies has revolutionized our understanding of diving behavior in marine taxa such as sharks, turtles, and seals during their wide-ranging movements. However, similar applications for large whales have lagged behind due to the difficulty of keeping tags on the animals for extended periods of time. Here, we present a novel configuration of a transdermally attached biologging device called the Advanced Dive Behavior (ADB) tag. The ADB tag contains sensors that record hydrostatic pressure, three-axis accelerometers, magnetometers, water temperature, and light level, all sampled at 1 Hz. The ADB tag also collects Fastloc GPS locations and can send dive summary data through Service Argos, while staying attached to a whale for typical periods of 3-7 weeks before releasing for recovery and subsequent data download. ADB tags were deployed on sperm whales ( Physeter macrocephalus; N  = 46), blue whales ( Balaenoptera musculus; N  = 8), and fin whales ( B. physalus; N  = 5) from 2007 to 2015, resulting in attachment durations from 0 to 49.6 days, and recording 31 to 2,539 GPS locations and 27 to 2,918 dives per deployment. Archived dive profiles matched well with published dive shapes of each species from short-term records. For blue and fin whales, feeding lunges were detected using peaks in accelerometer data and matched corresponding vertical excursions in the depth record. In sperm whales, rapid orientation changes in the accelerometer data, often during the bottom phase of dives, were likely related to prey pursuit, representing a relative measure of foraging effort. Sperm whales were documented repeatedly diving to, and likely foraging along, the seafloor. Data from the temperature sensor described the vertical structure of the water column in all three species, extending from the surface to depths >1,600 m. In addition to providing information needed to construct multiweek time budgets, the ADB tag is well suited to studying the effects of anthropogenic sound on whales by allowing for pre- and post-exposure monitoring of the whale's dive behavior. This tag begins to bridge the gap between existing long-duration but low-data throughput tags, and short-duration, high-resolution data loggers.

  15. Estimation of the radiation dose from radiotherapy for skin haemangiomas in childhood: the ICTA software for epidemiology

    NASA Astrophysics Data System (ADS)

    Shamsaldin, A.; Lundell, M.; Diallo, I.; Ligot, L.; Chavaudra, J.; de Vathaire, F.

    2000-12-01

    Radium applicators and pure beta emitters have been widely used in the past to treat skin haemangioma in early childhood. A well defined relationship between the low doses received from these applicators and radiation-induced cancers requires accurate dosimetry. A human-based CT scan phantom has been used to simulate every patient and treatment condition and then to calculate the source-target distance when radium and pure beta applicators were used. The effective transmission factor ϕ(r) for the gamma spectrum emitted by the radium sources applied on the skin surface was modelled using Monte Carlo simulations. The well-known quantization approach was used to calculate gamma doses delivered from radium applicators to various anatomical points. For 32P, 90Sr/90Y applicators and 90Y needles we have used the apparent exponential attenuation equation. The dose calculation algorithm was integrated into the ICTA software (standing for a model that constructs an Individualized phantom based on CT slices and Auxological data), which has been developed for epidemiological studies of cohorts of patients who received radium and beta-treatments for skin haemangioma. The ϕ(r) values obtained for radium skin applicators are in good agreement with the available values in the first 10 cm but higher at greater distances. Gamma doses can be calculated with this algorithm at 165 anatomical points throughout the body of patients treated with radium applicators. Lung heterogeneity and air crossed by the gamma rays are considered. Comparison of absorbed doses in water from a 10 mg equivalent radium source simulated by ICTA with those measured at the Radiumhemmet, Karolinska Hospital (RAH) showed good agreement, but ICTA estimation of organ doses did not always correspond those estimated at the RAH. Beta doses from 32P, 90Sr/90Y applicators and 90Y needles are calculated up to the maximum beta range (11 mm).

  16. Determining Beta Sheet Crystallinity in Fibrous Proteins by Thermal Analysis and Infrared Spectroscopy

    NASA Astrophysics Data System (ADS)

    Hu, Xiao; Kaplan, David; Cebe, Peggy

    2007-03-01

    We report a study of self-assembled beta pleated sheets in Bombyx mori silk fibroin films using thermal analysis and infrared spectroscopy. Crystallization of beta pleated sheets was effected either by heating the films above the glass transition temperature (Tg) and holding isothermally, or by exposure to methanol. The fractions of secondary structural components including random coils, alpha helices, beta pleated sheets, turns, and side chains, were evaluated using Fourier self-deconvolution (FSD) of the infrared absorbance spectra. As crystalline beta sheets form, the heat capacity increment from the TMDSC trace at Tg is systematically decreased and is linearly well correlated with beta sheet content determined from FSD. This analysis of beta sheet content can serve as an alternative to X-ray methods and may have wide applicability to other crystalline beta sheet forming proteins.

  17. Plan for DoD Wide Demonstrations of a DoD Improved Interactive Electronic Technical Manual (IETM) Architecture

    DTIC Science & Technology

    1998-07-01

    all the MS Word files into FrameMaker + SGML format and use the FrameMaker application to SGML tag all of the data in accordance with the Army TM...Document Type Definitions (DTDs) in MIL-STD- 2361. The edited SGML tagged files are saved as PDF files for delivery to the field. The FrameMaker ...as TIFF files and being imported into FrameMaker prior to saving the TMs as PDF files. Since the hardware to be used by the AN/PPS-5 technician is

  18. Process for the extemporaneous preparation of an injectable fatty acid tagged in the omega position by means of radioactive iodine

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bardy, A.; Comet, M.; Coornaert, S.

    1984-10-09

    A process is claimed for the preparation of a fatty acid tagged with radioactive iodine, where a brominated or iodized fatty acid is reacted, preferably in the omega position, with radioactive iodide in the dry state or with an aqueous solution of radioactive iodide, in the presence of vehicling iodide, to exchange the bromine or iodine of the fatty acid for radioactive iodine. Application to use as radio-pharmaceutical products for studying cardiac metabolism troubles in human beings by scintigraphy is mentioned.

  19. Multiport Circular Polarized RFID-Tag Antenna for UHF Sensor Applications.

    PubMed

    Zaid, Jamal; Abdulhadi, Abdulhadi; Kesavan, Arun; Belaizi, Yassin; Denidni, Tayeb A

    2017-07-05

    A circular polarized patch antenna for UHF RFID tag-based sensor applications is presented, with the circular polarization (CP) generated by a new antenna shape, an asymmetric stars shaped slotted microstrip patch antenna (CP-ASSSMP). Four stars etched on the patch allow the antenna's size to be reduced by close to 20%. The proposed antenna is matched with two RFID chips via inductive-loop matching. The first chip is connected to a resistive sensor and acts as a sensor node, and the second is used as a reference node. The proposed antenna is used for two targets, serving as both reference and sensor simultaneously, thereby eliminating the need for a second antenna. Its reader can read the RFID chips at any orientation of the tag due to the CP. The measured reading range is about 25 m with mismatch polarization. The operating frequency band is 902-929 MHz for the two ports, which is covered by the US RFID band, and the axial-ratio bandwidth is about 7 MHz. In addition, the reader can also detect temperature, based on the minimum difference in the power required by the reference and sensor.

  20. Multiport Circular Polarized RFID-Tag Antenna for UHF Sensor Applications

    PubMed Central

    Zaid, Jamal; Abdulhadi, Abdulhadi; Kesavan, Arun; Belaizi, Yassin; Denidni, Tayeb A.

    2017-01-01

    A circular polarized patch antenna for UHF RFID tag-based sensor applications is presented, with the circular polarization (CP) generated by a new antenna shape, an asymmetric stars shaped slotted microstrip patch antenna (CP-ASSSMP). Four stars etched on the patch allow the antenna’s size to be reduced by close to 20%. The proposed antenna is matched with two RFID chips via inductive-loop matching. The first chip is connected to a resistive sensor and acts as a sensor node, and the second is used as a reference node. The proposed antenna is used for two targets, serving as both reference and sensor simultaneously, thereby eliminating the need for a second antenna. Its reader can read the RFID chips at any orientation of the tag due to the CP. The measured reading range is about 25 m with mismatch polarization. The operating frequency band is 902–929 MHz for the two ports, which is covered by the US RFID band, and the axial-ratio bandwidth is about 7 MHz. In addition, the reader can also detect temperature, based on the minimum difference in the power required by the reference and sensor. PMID:28678178

  1. The effects of aromatic amino acid derivatives on the excitability of an identifiable giant neurone of the African giant snail (Achatina fulica Férussac).

    PubMed Central

    Takeuchi, H.; Tamura, H.

    1980-01-01

    1 The effects of derivatives of aromatic amino acids on the excitability of an identifiable giant neurone (TAN, tonically autoactive neurone) of the African giant snail (Achatina fulica Férussac) were examined. 2 The following substances had marked inhibitory effects on TAN using bath application: N-beta-phenylpropionyl-L-Tyr and N-beta-phenylpropionyl-L-Trp (critical concentration, 3 x 10(-7) M), N-beta-phenylpropionyl-L-Phe, N-cinnamoyl-DL-Trp and N-phenoxyacetyl-L-Trp (critical concentration, 10(-5) to 3 x 10(-5) M). However, N-beta-phenylpropionyl-D-Tyr and N-beta-phenylpropionyl tyramine had no effect. 3 Microdrop (150 micrometers in diameter) application of N-beta-phenylpropionyl-L-Tyr or N-beta-phenylpropionyl-l-trp containing about 100 pg resulted in marked inhibitory effects on TAN. The effect was observed in Ca2+-free, Mg2+-rich (24 mM) solution. Substitution of Cl- by acetate did not alter the response. This indicates that the two substances act directly on the TAN membrane and not via synaptic influences, and that the inhibition produced by the two substances is not due to the permeability increase of the TAN membrane to Cl-. PMID:7378654

  2. ReAsH/FlAsH labeling and image analysis of tetracysteine sensor proteins in cells.

    PubMed

    Irtegun, Sevgi; Ramdzan, Yasmin M; Mulhern, Terrence D; Hatters, Danny M

    2011-08-31

    Fluorescent proteins and dyes are essential tools for the study of protein trafficking, localization and function in cells. While fluorescent proteins such as green fluorescence protein (GFP) have been extensively used as fusion partners to proteins to track the properties of a protein of interest, recent developments with smaller tags enable new functionalities of proteins to be examined in cells such as conformational change and protein-association. One small tag system involves a tetracysteine motif (CCXXCC) genetically inserted into a target protein, which binds to biarsenical dyes, ReAsH (red fluorescent) and FlAsH (green fluorescent), with high specificity even in live cells. The TC/biarsenical dye system offers far less steric constraints to the host protein than fluorescent proteins which has enabled several new approaches to measure conformational change and protein-protein interactions. We recently developed a novel application of TC tags as sensors of oligomerization in cells expressing mutant huntingtin, which when mutated aggregates in neurons in Huntington disease. Huntingtin was tagged with two fluorescent dyes, one a fluorescent protein to track protein location, and the second a TC tag which only binds biarsenical dyes in monomers. Hence, changes in colocalization between protein and biarsenical dye reactivity enabled submicroscopic oligomer content to be spatially mapped within cells. Here, we describe how to label TC-tagged proteins fused to a fluorescent protein (Cherry, GFP or CFP) with FlAsH or ReAsH in live mammalian cells and how to quantify the two color fluorescence (Cherry/FlAsH, CFP/FlAsH or GFP/ReAsH combinations).

  3. Application of Strep-Tactin XT for affinity purification of Twin-Strep-tagged CB2, a G protein-coupled cannabinoid receptor

    PubMed Central

    Yeliseev, Alexei; Zoubak, Lioudmila; Schmidt, Thomas G.M.

    2017-01-01

    Human cannabinoid receptor CB2 belongs to the class A of G protein-coupled receptor (GPCR). High resolution structural studies of CB2 require milligram quantities of purified, structurally intact protein. Here we describe an efficient protocol for purification of this protein using the Twin-Strep-tag/Strep-Tactin XT system. To improve the affinity of interaction of the recombinant CB2 with the resin, the double repeat of the Strep-tag was attached either to the N- or C-terminus of CB2 via a short linker. The CB2 was isolated at high purity from dilute solutions containing high concentrations of detergents, glycerol and salts, by capturing onto the Strep-Tactin XT resin, and was eluted from the resin under mild conditions upon addition of biotin. Surface plasmon resonance studies performed demonstrate the high affinity of interaction between the Twin-Strep-tag fused to the CB2 and Strep-Tactin XT with an estimated Kd in the low nanomolar range. The affinity of binding did not vary significantly in response to the position of the tag at either N- or C-termini of the fusion. The variation in the length of the linker between the double repeats of the Strep-tag from 6 to 12 amino acid residues did not significantly affect the binding. The novel purification protocol reported here enables efficient isolation of a recombinant GPCR expressed at low titers in host cells. This procedure is suitable for preparation of milligram quantities of stable isotope-labelled receptor for high-resolution NMR studies. PMID:27867058

  4. Authentication via wavefront-shaped optical responses

    NASA Astrophysics Data System (ADS)

    Eilers, Hergen; Anderson, Benjamin R.; Gunawidjaja, Ray

    2018-02-01

    Authentication/tamper-indication is required in a wide range of applications, including nuclear materials management and product counterfeit detection. State-of-the-art techniques include reflective particle tags, laser speckle authentication, and birefringent seals. Each of these passive techniques has its own advantages and disadvantages, including the need for complex image comparisons, limited flexibility, sensitivity to environmental conditions, limited functionality, etc. We have developed a new active approach to address some of these short-comings. The use of an active characterization technique adds more flexibility and additional layers of security over current techniques. Our approach uses randomly-distributed nanoparticles embedded in a polymer matrix (tag/seal) which is attached to the item to be secured. A spatial light modulator is used to adjust the wavefront of a laser which interacts with the tag/seal, and a detector is used to monitor this interaction. The interaction can occur in various ways, including transmittance, reflectance, fluorescence, random lasing, etc. For example, at the time of origination, the wavefront-shaped reflectance from a tag/seal can be adjusted to result in a specific pattern (symbol, words, etc.) Any tampering with the tag/seal would results in a disturbance of the random orientation of the nanoparticles and thus distort the reflectance pattern. A holographic waveplate could be inserted into the laser beam for verification. The absence/distortion of the original pattern would then indicate that tampering has occurred. We have tested the tag/seal's and authentication method's tamper-indicating ability using various attack methods, including mechanical, thermal, and chemical attacks, and have verified our material/method's robust tamper-indicating ability.

  5. DNA-tagged Microparticles for Tracing Water Flows and Travel Times in Natural Systems: The First results from Controlled Laboratory Experiments

    NASA Astrophysics Data System (ADS)

    Bogaard, T.; Bandyopadhyay, S.; Foppen, J. W.

    2017-12-01

    Societal demand for water safety is continuously increasing, being it resilient against flood/droughts, clean water for ecosystems, recreation or safe drinking water. Robust methods to measure temporal and spatial patterns of water and contaminant pathways are still lacking. Our research project aims to develop and apply (1) innovative, robust, and environmental-friendly silica-protected iron oxide micro-particles tagged with artificial DNA to trace contaminant movement and travel times of water in natural systems and (2) an innovative coupled model approach to capture dynamics in hydrological pathways and their effects on water quality. The exceptional property of DNA-tagging is the infinite number of unique tracers that can be produced and their detectability at extreme low concentrations. The advantage of the iron-core of the particle is the magnetic harvesting of the particles from water-samples. Such tracers are thought to give the water sector a unique tool for in-situ mapping of transport of contaminants and pathogenic microorganisms in water systems. However, the characteristics of the particle like magnetic property of the iron-core and surface potential of the silica layer, are of key importance for the behaviour of the particle in surface water and in soils. Furthermore, the application of such micro-particles requires strict protocols for the experiment, sampling and laboratory handling which are currently not available. We used two different types of silica-protected DNA-tagged micro-particles. We performed batch, column and flow experiments to assess the behaviour of the particles. We will present the first results of the controlled laboratory experiments for hydrological tracing. We will discuss the results and link it to the differences in particles design. Furthermore, we will draw conclusions and discuss knowledge gaps for future application of silica-protected DNA-tagged micro-particles in hydrological research.

  6. Application of a New Dual Localization-Affinity Purification Tag Reveals Novel Aspects of Protein Kinase Biology in Aspergillus nidulans

    PubMed Central

    De Souza, Colin P.; Hashmi, Shahr B.; Osmani, Aysha H.; Osmani, Stephen A.

    2014-01-01

    Filamentous fungi occupy critical environmental niches and have numerous beneficial industrial applications but devastating effects as pathogens and agents of food spoilage. As regulators of essentially all biological processes protein kinases have been intensively studied but how they regulate the often unique biology of filamentous fungi is not completely understood. Significant understanding of filamentous fungal biology has come from the study of the model organism Aspergillus nidulans using a combination of molecular genetics, biochemistry, cell biology and genomic approaches. Here we describe dual localization-affinity purification (DLAP) tags enabling endogenous N or C-terminal protein tagging for localization and biochemical studies in A. nidulans. To establish DLAP tag utility we endogenously tagged 17 protein kinases for analysis by live cell imaging and affinity purification. Proteomic analysis of purifications by mass spectrometry confirmed association of the CotA and NimXCdk1 kinases with known binding partners and verified a predicted interaction of the SldABub1/R1 spindle assembly checkpoint kinase with SldBBub3. We demonstrate that the single TOR kinase of A. nidulans locates to vacuoles and vesicles, suggesting that the function of endomembranes as major TOR cellular hubs is conserved in filamentous fungi. Comparative analysis revealed 7 kinases with mitotic specific locations including An-Cdc7 which unexpectedly located to mitotic spindle pole bodies (SPBs), the first such localization described for this family of DNA replication kinases. We show that the SepH septation kinase locates to SPBs specifically in the basal region of apical cells in a biphasic manner during mitosis and again during septation. This results in gradients of SepH between G1 SPBs which shift along hyphae as each septum forms. We propose that SepH regulates the septation initiation network (SIN) specifically at SPBs in the basal region of G1 cells and that localized gradients of SIN activity promote asymmetric septation. PMID:24599037

  7. Application of a new dual localization-affinity purification tag reveals novel aspects of protein kinase biology in Aspergillus nidulans.

    PubMed

    De Souza, Colin P; Hashmi, Shahr B; Osmani, Aysha H; Osmani, Stephen A

    2014-01-01

    Filamentous fungi occupy critical environmental niches and have numerous beneficial industrial applications but devastating effects as pathogens and agents of food spoilage. As regulators of essentially all biological processes protein kinases have been intensively studied but how they regulate the often unique biology of filamentous fungi is not completely understood. Significant understanding of filamentous fungal biology has come from the study of the model organism Aspergillus nidulans using a combination of molecular genetics, biochemistry, cell biology and genomic approaches. Here we describe dual localization-affinity purification (DLAP) tags enabling endogenous N or C-terminal protein tagging for localization and biochemical studies in A. nidulans. To establish DLAP tag utility we endogenously tagged 17 protein kinases for analysis by live cell imaging and affinity purification. Proteomic analysis of purifications by mass spectrometry confirmed association of the CotA and NimXCdk1 kinases with known binding partners and verified a predicted interaction of the SldABub1/R1 spindle assembly checkpoint kinase with SldBBub3. We demonstrate that the single TOR kinase of A. nidulans locates to vacuoles and vesicles, suggesting that the function of endomembranes as major TOR cellular hubs is conserved in filamentous fungi. Comparative analysis revealed 7 kinases with mitotic specific locations including An-Cdc7 which unexpectedly located to mitotic spindle pole bodies (SPBs), the first such localization described for this family of DNA replication kinases. We show that the SepH septation kinase locates to SPBs specifically in the basal region of apical cells in a biphasic manner during mitosis and again during septation. This results in gradients of SepH between G1 SPBs which shift along hyphae as each septum forms. We propose that SepH regulates the septation initiation network (SIN) specifically at SPBs in the basal region of G1 cells and that localized gradients of SIN activity promote asymmetric septation.

  8. Beta-Testing Agreement | Frederick National Laboratory for Cancer Research

    Cancer.gov

    Beta-Testing Agreements are appropriate forlimited term evaluation and applications development of new software, technology, or equipment platforms by the Frederick National Laboratory in collaboration with an external commercial partner. It ma

  9. Guidance of neurite outgrowth on aligned electrospun polypyrrole/poly(styrene-beta-isobutylene-beta-styrene) fiber platforms.

    PubMed

    Liu, Xiao; Chen, Jun; Gilmore, Kerry J; Higgins, Michael J; Liu, Yong; Wallace, Gordon G

    2010-09-15

    The purpose of this work was to investigate the potential biomedical application of novel aligned electrospun polypyrrole (PPy)/poly(styrene-beta-isobutylene-beta-styrene) (SIBS) fibers. After successfully aligning the electroactive PPy/SIBS fibers based on our modified electrospinning method, we demonstrated that neurite outgrowth from PC12 cells could be highly orientated parallel to the aligned PPy/SIBS fibers. Physical interactions between the nerve cells and PPy/SIBS fibers through filopodia "sensing" were observed using atomic force microscopy. These observations indicate a role of contact guidance as a mechanism for the observed alignment. This work highlights the capacity for electroactive PPy/SIBS fibers to support and guide nerve cell differentiation through topographic cues, which is a highly desirable characteristic in medical implants for neurological applications. (c) 2010 Wiley Periodicals, Inc.

  10. 75 FR 70236 - Hop Beta Acids; Receipt of Application for Emergency Exemption, Solicitation of Public Comment

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-17

    ... of Agriculture to use hop beta acids (CAS Reg. No. none specified) to treat up to 181,000 honey bee... exemption regional request for use of hop beta acids in honey bee hives to control varroa mites. Information... effect on honey bee populations. The parasitic mite is considered the primary pest of honeybees and its...

  11. Spatial Double Generalized Beta Regression Models: Extensions and Application to Study Quality of Education in Colombia

    ERIC Educational Resources Information Center

    Cepeda-Cuervo, Edilberto; Núñez-Antón, Vicente

    2013-01-01

    In this article, a proposed Bayesian extension of the generalized beta spatial regression models is applied to the analysis of the quality of education in Colombia. We briefly revise the beta distribution and describe the joint modeling approach for the mean and dispersion parameters in the spatial regression models' setting. Finally, we motivate…

  12. Characterization of a new multifunctional beta-glucosidase from Musca domestica.

    PubMed

    Zhang, Shu; Huang, Jian; Hu, Rong; Guo, Guo; Shang, Xiaoli; Wu, Jianwei

    2017-08-01

    To engineer Pichia pastoris for heterologous production of cellulase from Musca domestica and explore its potential for industrial applications. A new beta-glucosidase gene (bg), encoding 562 amino acids, was cloned from M. domestica by using rapid amplification of cDNA ends. The gene bg was linked to pPICZαA and expressed in P. pastoris with a yield of 500 mg l -1 . The enzyme has the maximum activity with 27.6 U mg -1 towards cellulose. The beta-glucosidase has stable activity from 20 to 70 °C and can tolerate one-mole glucose. It has the maximum activities for salicin (25.9 ± 1.8 U mg -1 ), cellobiose (40.1 ± 2.3 U mg -1 ) and cellulose (27.6 ± 3.5 U mg -1 ). The wide-range substrate activities of the beta-glucosidase were further verified by matrix-assisted laser desorption/ionization mass spectra. Structural analysis shows that the beta-glucosidase belongs to glycoside hydrolase family Ι and possesses O-glycosylation sites. Thus, a multifunctional beta-glucosidase was expressed from M. domestica and provides a potential tool for industrial application of cellulose.

  13. Polyesters from microorganisms.

    PubMed

    Kim, Y B; Lenz, R W

    2001-01-01

    Bacterial polyesters have been found to have useful properties for applications as thermoplastics, elastomers, and adhesives and are biodegradable and biocompatible. Poly(3-hydroxyalkanoates) (PHAs) and poly(beta-malate) are the most representative polyesters synthesized by microorganisms. PHAs containing a wide variety of repeating units can be produced by bacteria, including those containing many types of pendant functional groups which can be synthesized by microorganisms that are grown on unnatural organic substrates. Poly(beta-malate) is of interest primarily for medical applications, especially for drug delivery systems. In this chapter, the bacterial production and properties of poly(3-hydroxyalkanoates) and poly(beta-malate) are described with emphasis on the former.

  14. Interoperation of an UHF RFID Reader and a TCP/IP Device via Wired and Wireless Links

    PubMed Central

    Lee, Sang Hoon; Jin, Ik Soo

    2011-01-01

    A main application in radio frequency identification (RFID) sensor networks is the function that processes real-time tag information after gathering the required data from multiple RFID tags. The component technologies that contain an RFID reader, called the interrogator, which has a tag chip, processors, coupling antenna, and a power management system have advanced significantly over the last decade. This paper presents a system implementation for interoperation between an UHF RFID reader and a TCP/IP device that is used as a gateway. The proposed system consists of an UHF RFID tag, an UHF RFID reader, an RF end-device, an RF coordinator, and a TCP/IP I/F. The UHF RFID reader, operating at 915 MHz, is compatible with EPC Class-0/Gen1, Class-1/Gen1 and 2, and ISO18000-6B. In particular, the UHF RFID reader can be combined with the RF end-device/coordinator for a ZigBee (IEEE 802.15.4) interface, which is a low-power wireless standard. The TCP/IP device communicates with the RFID reader via wired links. On the other hand, it is connected to the ZigBee end-device via wireless links. The web based test results show that the developed system can remotely recognize information of multiple tags through the interoperation between the RFID reader and the TCP/IP device. PMID:22346665

  15. Interoperation of an UHF RFID reader and a TCP/IP device via wired and wireless links.

    PubMed

    Lee, Sang Hoon; Jin, Ik Soo

    2011-01-01

    A main application in radio frequency identification (RFID) sensor networks is the function that processes real-time tag information after gathering the required data from multiple RFID tags. The component technologies that contain an RFID reader, called the interrogator, which has a tag chip, processors, coupling antenna, and a power management system have advanced significantly over the last decade. This paper presents a system implementation for interoperation between an UHF RFID reader and a TCP/IP device that is used as a gateway. The proposed system consists of an UHF RFID tag, an UHF RFID reader, an RF end-device, an RF coordinator, and a TCP/IP I/F. The UHF RFID reader, operating at 915 MHz, is compatible with EPC Class-0/Gen1, Class-1/Gen1 and 2, and ISO18000-6B. In particular, the UHF RFID reader can be combined with the RF end-device/coordinator for a ZigBee (IEEE 802.15.4) interface, which is a low-power wireless standard. The TCP/IP device communicates with the RFID reader via wired links. On the other hand, it is connected to the ZigBee end-device via wireless links. The web based test results show that the developed system can remotely recognize information of multiple tags through the interoperation between the RFID reader and the TCP/IP device.

  16. Tagging of Test Tubes with Electronic p-Chips for Use in Biorepositories.

    PubMed

    Mandecki, Wlodek; Kopacka, Wesley M; Qian, Ziye; Ertwine, Von; Gedzberg, Katie; Gruda, Maryann; Reinhardt, David; Rodriguez, Efrain

    2017-08-01

    A system has been developed to electronically tag and track test tubes used in biorepositories. The system is based on a light-activated microtransponder, also known as a "p-Chip." One of the pressing problems with storing and retrieving biological samples at low temperatures is the difficulty of reliably reading the identification (ID) number that links each storage tube with the database containing sample details. Commonly used barcodes are not always reliable at low temperatures because of poor adhesion of the label to the test tube and problems with reading under conditions of frost and ice accumulation. Traditional radio frequency identification (RFID) tags are not cost effective and are too large for this application. The system described herein consists of the p-Chip, p-Chip-tagged test tubes, two ID readers (for single tubes or for racks of tubes), and software. We also describe a robot that is configured for retrofitting legacy test tubes in biorepositories with p-Chips while maintaining the temperature of the sample below -50°C at all times. The main benefits of the p-Chip over other RFID devices are its small size (600 × 600 × 100 μm) that allows even very small tubes or vials to be tagged, low cost due to the chip's unitary construction, durability, and the ability to read the ID through frost and ice.

  17. Viral infection of the marine alga Emiliania huxleyi triggers lipidome remodeling and induces the production of highly saturated triacylglycerol.

    PubMed

    Malitsky, Sergey; Ziv, Carmit; Rosenwasser, Shilo; Zheng, Shuning; Schatz, Daniella; Porat, Ziv; Ben-Dor, Shifra; Aharoni, Asaph; Vardi, Assaf

    2016-04-01

    Viruses that infect marine photosynthetic microorganisms are major ecological and evolutionary drivers of microbial food webs, estimated to turn over more than a quarter of the total photosynthetically fixed carbon. Viral infection of the bloom-forming microalga Emiliania huxleyi induces the rapid remodeling of host primary metabolism, targeted towards fatty acid metabolism. We applied a liquid chromatography-mass spectrometry (LC-MS)-based lipidomics approach combined with imaging flow cytometry and gene expression profiling to explore the impact of viral-induced metabolic reprogramming on lipid composition. Lytic viral infection led to remodeling of the cellular lipidome, by predominantly inducing the biosynthesis of highly saturated triacylglycerols (TAGs), coupled with a significant accumulation of neutral lipids within lipid droplets. Furthermore, TAGs were found to be a major component (77%) of the lipidome of isolated virions. Interestingly, viral-induced TAGs were significantly more saturated than TAGs produced under nitrogen starvation. This study highlights TAGs as major products of the viral-induced metabolic reprogramming during the host-virus interaction and indicates a selective mode of membrane recruitment during viral assembly, possibly by budding of the virus from specialized subcellular compartments. These findings provide novel insights into the role of viruses infecting microalgae in regulating metabolism and energy transfer in the marine environment and suggest their possible biotechnological application in biofuel production. © 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.

  18. Molecular imaging and sensing using plasmonic nanoparticles

    NASA Astrophysics Data System (ADS)

    Crow, Matthew James

    Noble metal nanoparticles exhibit unique optical properties that are beneficial to a variety of applications, including molecular imaging. The large scattering cross sections of nanoparticles provide high contrast necessary for biomarkers. Unlike alternative contrast agents, nanoparticles provide refractive index sensitivity revealing information regarding the local cellular environment. Altering the shape and composition of the nanoparticle shifts the peak resonant wavelength of scattered light, allowing for implementation of multiple spectrally distinct tags. In this project, nanoparticles that scatter in different spectral windows are functionalized with various antibodies recognizing extra-cellular receptors integral to cancer progression. A hyperspectral imaging system is developed, allowing for visualization and spectral characterization of cells labeled with these conjugates. Various molecular imaging and microspectroscopy applications of plasmonic nanoparticles are then investigated. First, anti-EGFR gold nanospheres are shown to quantitatively measure receptor expression with similar performance to fluorescence assays. Second, anti-EGFR gold nanorods and novel anti-IGF-1R silver nanospheres are implemented to indicate local cellular refractive indices. Third, because biosensing capabilities of nanoparticle tags may be limited by plasmonic coupling, polarization mapping is investigated as a method to discern these effects. Fourth, plasmonic coupling is tested to monitor HER-2 dimerization. Experiments reveal the interparticle conformation of proximal HER-2 bound labels, required for plasmonic coupling-enhanced dielectric sensing. Fifth, all three functionalized plasmonic tags are implemented simultaneously to indicate clinically relevant cell immunophenotype information and changes in the cellular dielectric environment. Finally, flow cytometry experiments are conducted utilizing the anti-EGFR nanorod tag to demonstrate profiling of receptor expression distribution and potential increased multiplexing capability.

  19. The electrophysiologic properties of esmolol, a short acting beta-blocker.

    PubMed

    Greenspan, A M; Spielman, S R; Horowitz, L N; Laddu, A; Senior, S

    1988-04-01

    Although beta-blockers have established efficacy in treating ventricular ectopy and PSVT, their applicability for acute antiarrhythmic interventions in patients with organic heart disease or COPD, is frequently limited by negative inotropic or bronchospastic side effects. The development of an ultrashort acting beta-blocker with rapid reversibility of its side effects would widen their applicability. Therefore, we tested the electrophysiologic properties of such a new short acting beta-blocker, esmolol, in 14 patients (10 with organic heart disease) with a mean EF of 47.6 +/- 17%, undergoing standard clinical electrophysiologic studies for various indications. Like most other beta-blockers, esmolol's major direct effects were on sinus node function and AV nodal conduction characteristics; significantly prolonging sinus cycle length, cycle length to Wenckebach and AH interval in sinus rhythm and at a paced cycle length of 600 ms. In contrast to most other beta-blockers, following termination of its infusion, esmolol shortened parameters of sinus node function and AV nodal refractoriness, with respect to the control values, suggesting a possible rebound phenomena. These effects occurred within 5 min of terminating the intravenous drug infusion. Esmolol had no significant effect on systolic blood pressure, electrocardiographic intervals and had rare adverse reactions. We conclude that esmolol is an ultra-short acting beta-blocker, with typical direct electrophysiologic effects on sinus node and AV nodal function, and a possible rebound phenomena following its discontinuation that may make it particularly suited to acute antiarrhythmic interventions in patients susceptible to adverse beta-blocker side effects.

  20. Monitoring and Evaluation: Statistical Support for Life-cycle Studies, Annual Report 2003.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Skalski, John

    2003-11-01

    The ongoing mission of this project is the development of statistical tools for analyzing fisheries tagging data in the most precise and appropriate manner possible. This mission also includes providing statistical guidance on the best ways to design large-scale tagging studies. This mission continues because the technologies for conducting fish tagging studies continuously evolve. In just the last decade, fisheries biologists have seen the evolution from freeze-brands and coded wire tags (CWT) to passive integrated transponder (PIT) tags, balloon-tags, radiotelemetry, and now, acoustic-tags. With each advance, the technology holds the promise of more detailed and precise information. However, the technologymore » for analyzing and interpreting the data also becomes more complex as the tagging techniques become more sophisticated. The goal of the project is to develop the analytical tools in parallel with the technical advances in tagging studies, so that maximum information can be extracted on a timely basis. Associated with this mission is the transfer of these analytical capabilities to the field investigators to assure consistency and the highest levels of design and analysis throughout the fisheries community. Consequently, this project provides detailed technical assistance on the design and analysis of tagging studies to groups requesting assistance throughout the fisheries community. Ideally, each project and each investigator would invest in the statistical support needed for the successful completion of their study. However, this is an ideal that is rarely if every attained. Furthermore, there is only a small pool of highly trained scientists in this specialized area of tag analysis here in the Northwest. Project 198910700 provides the financial support to sustain this local expertise on the statistical theory of tag analysis at the University of Washington and make it available to the fisheries community. Piecemeal and fragmented support from various agencies and organizations would be incapable of maintaining a center of expertise. The mission of the project is to help assure tagging studies are designed and analyzed from the onset to extract the best available information using state-of-the-art statistical methods. The overarching goals of the project is to assure statistically sound survival studies so that fish managers can focus on the management implications of their findings and not be distracted by concerns whether the studies are statistically reliable or not. Specific goals and objectives of the study include the following: (1) Provide consistent application of statistical methodologies for survival estimation across all salmon life cycle stages to assure comparable performance measures and assessment of results through time, to maximize learning and adaptive management opportunities, and to improve and maintain the ability to responsibly evaluate the success of implemented Columbia River FWP salmonid mitigation programs and identify future mitigation options. (2) Improve analytical capabilities to conduct research on survival processes of wild and hatchery chinook and steelhead during smolt outmigration, to improve monitoring and evaluation capabilities and assist in-season river management to optimize operational and fish passage strategies to maximize survival. (3) Extend statistical support to estimate ocean survival and in-river survival of returning adults. Provide statistical guidance in implementing a river-wide adult PIT-tag detection capability. (4) Develop statistical methods for survival estimation for all potential users and make this information available through peer-reviewed publications, statistical software, and technology transfers to organizations such as NOAA Fisheries, the Fish Passage Center, US Fish and Wildlife Service, US Geological Survey (USGS), US Army Corps of Engineers (USACE), Public Utility Districts (PUDs), the Independent Scientific Advisory Board (ISAB), and other members of the Northwest fisheries community. (5) Provide and maintain statistical software for tag analysis and user support. (6) Provide improvements in statistical theory and software as requested by user groups. These improvements include extending software capabilities to address new research issues, adapting tagging techniques to new study designs, and extending the analysis capabilities to new technologies such as radio-tags and acoustic-tags.« less

  1. Radon-related backgrounds in the LUX dark matter search

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bradley, A.; Akerib, D. S.; Araújo, H. M.

    The LUX detector is currently in operation at the Davis Campus at the 4850’ level of the Sanford Underground Research Facility (SURF) in Lead, SD to directly search for WIMP dark matter. Knowing the type and rate of backgrounds is critical in a rare, low energy event search, and LUX was designed, constructed, and deployed to mitigate backgrounds, both internal and external. An important internal background are decays of radon and its daughters. These consist of alpha decays, which are easily tagged and are a tracer of certain backgrounds, and beta decays, some of which are not as readily taggedmore » and present a background for the WIMP search. We report on studies of alpha decay and discuss implications for the WIMP search.« less

  2. Radon-related backgrounds in the LUX dark matter search

    DOE PAGES

    Bradley, A.; Akerib, D. S.; Araújo, H. M.; ...

    2015-01-01

    The LUX detector is currently in operation at the Davis Campus at the 4850’ level of the Sanford Underground Research Facility (SURF) in Lead, SD to directly search for WIMP dark matter. Knowing the type and rate of backgrounds is critical in a rare, low energy event search, and LUX was designed, constructed, and deployed to mitigate backgrounds, both internal and external. An important internal background are decays of radon and its daughters. These consist of alpha decays, which are easily tagged and are a tracer of certain backgrounds, and beta decays, some of which are not as readily taggedmore » and present a background for the WIMP search. We report on studies of alpha decay and discuss implications for the WIMP search.« less

  3. Use of radio frequency identification (RFID) tags in bedside monitoring of endotracheal tube position.

    PubMed

    Reicher, Joshua; Reicher, Danielle; Reicher, Murray

    2007-06-01

    Improper positioning of the endotracheal tube during intubation poses a serious health risk to patients. In one prospective study of 219 critically ill patients, 14% required endotracheal tube repositioning after intubation [Brunel et al. Chest 1989; 96: 1043-1045] While a variety of techniques are used to confirm proper tube placement, a chest X-ray is usually employed for definitive verification. Radio frequency identification (RFID) technology, in which an RFID reader emits and receives a signal from an RFID tag, may be useful in evaluating endotracheal tube position. RFID technology has already been approved for use in humans as a safe and effective tool in a variety of applications. The use of handheld RFID detectors and RFID tag-labeled endotracheal tubes could allow for easy and accurate bedside monitoring of endotracheal tube position, once initial proper placement is confirmed.

  4. Coplanar UHF RFID tag antenna with U-shaped inductively coupled feed for metallic applications.

    PubMed

    Salman, Karrar Naji; Ismail, Alyani; Raja Abdullah, Raja Syamsul Azmir; Saeedi, Tale

    2017-01-01

    In this paper, we present a novel compact, coplanar, tag antenna design for metallic objects. Electrically small antenna has designed for a UHF RFID (860-960 MHz) based on a proximity-coupled feed through. Furthermore, two symmetrical Via-loaded coplanar grounds fed by a U-shaped inductively coupled feed through an embedded transmission line. This configuration results in an antenna with dimensions of 31 × 19.5 × 3.065 mm3 at 915 MHz, and the total gain for the antenna is 0.12 dBi. The Via-loaded coplanar and U-shaped inductively coupled feeds allow the antenna to provide flexible tuning in terms of antenna impedance. In addition, a figure of merit is applied for the proposed tag antenna, and the results are presented. The read range is measured to be 4.2 m, which is very close to simulated values. This antenna measurement shows very good agreement with simulations.

  5. Behavior-Based Cleaning for Unreliable RFID Data Sets

    PubMed Central

    Fan, Hua; Wu, Quanyuan; Lin, Yisong

    2012-01-01

    Radio Frequency IDentification (RFID) technology promises to revolutionize the way we track items and assets, but in RFID systems, missreading is a common phenomenon and it poses an enormous challenge to RFID data management, so accurate data cleaning becomes an essential task for the successful deployment of systems. In this paper, we present the design and development of a RFID data cleaning system, the first declarative, behavior-based unreliable RFID data smoothing system. We take advantage of kinematic characteristics of tags to assist in RFID data cleaning. In order to establish the conversion relationship between RFID data and kinematic parameters of the tags, we propose a movement behavior detection model. Moreover, a Reverse Order Filling Mechanism is proposed to ensure a more complete access to get the movement behavior characteristics of tag. Finally, we validate our solution with a common RFID application and demonstrate the advantages of our approach through extensive simulations. PMID:23112595

  6. Behavior-based cleaning for unreliable RFID data sets.

    PubMed

    Fan, Hua; Wu, Quanyuan; Lin, Yisong

    2012-01-01

    Radio Frequency IDentification (RFID) technology promises to revolutionize the way we track items and assets, but in RFID systems, missreading is a common phenomenon and it poses an enormous challenge to RFID data management, so accurate data cleaning becomes an essential task for the successful deployment of systems. In this paper, we present the design and development of a RFID data cleaning system, the first declarative, behavior-based unreliable RFID data smoothing system. We take advantage of kinematic characteristics of tags to assist in RFID data cleaning. In order to establish the conversion relationship between RFID data and kinematic parameters of the tags, we propose a movement behavior detection model. Moreover, a Reverse Order Filling Mechanism is proposed to ensure a more complete access to get the movement behavior characteristics of tag. Finally, we validate our solution with a common RFID application and demonstrate the advantages of our approach through extensive simulations.

  7. Passive RFID Localisation Framework in Smart Homes Healthcare Settings.

    PubMed

    Alsinglawi, Belal; Liu, Tony; Nguyen, Quang Vinh; Gunawardana, Upul; Maeder, Anthony; Simoff, Simeon

    2016-01-01

    In recent years, Smart Homes have become a solution to benefit impaired individuals and elderly in their daily life settings. In healthcare applications, pervasive technologies have enabled the practicality of personal monitoring using Indoor positioning technologies. Radio-Frequency Identification (RFID) is a promising technology, which is useful for non-invasive tracking of activities of daily living. Many implementations have focused on using battery-enabled tags like in RFID active tags, which require frequent maintenance and they are costly. Other systems can use wearable sensors requiring individuals to wear tags which may be inappropriate for elders. Successful implementations of a tracking system are dependent on multiple considerations beyond the physical performance of the solution, such as affordability and human acceptance. This paper presents a localisation framework using passive RFID sensors. It aims to provide a low cost solution for subject location in Smart Homes healthcare.

  8. Flexible ultra-wideband antenna incorporated with metamaterial structures: multiple notches for chipless RFID application

    NASA Astrophysics Data System (ADS)

    Jalil, M. E.; Rahim, M. K. A.; Samsuri, N. A.; Dewan, R.; Kamardin, K.

    2017-01-01

    A coplanar waveguide (CPW) ultra-wideband (UWB) antenna incorporated with metamaterial—split ring resonator structure—that operates from 3.0 to 12.0 GHz is proposed for chipless RFID tag. The 30 mm × 40 mm flexible chipless RFID tag is designed on the fleece substrate ( ɛ r = 1.35, thickness = 1 mm and tan δ = 0.025). A six-slotted modified complementary split ring resonator (MCSRR) is introduced into the ultra-wideband antenna to produce multiple band notches at 3.0, 4.0, 5.0, 6.0 and 7.0 GHz. The frequency shifting technique is introduced for designing a high-capacity chipless RFID tag with compact size. Each MCSRR is able to code in four different allocations (00, 01, 10 and 11). To achieve encoding of 10-bits data (10,234 number), six MCSRRs are proposed with three-slotted MCSRR in the radiator and three-slotted MCSRR in the ground plane.

  9. 29 CFR 1915.80 - Scope, application, definitions, and effective dates.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... that provides an impediment (in addition to the energy-isolating device) to the release of energy or..., or system under a lockout/tags-plus application. (4) Capable of being locked out. An energy-isolating... other means of attachment to which, or through which, a lock can be affixed. Other energy-isolating...

  10. 29 CFR 1915.80 - Scope, application, definitions, and effective dates.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... that provides an impediment (in addition to the energy-isolating device) to the release of energy or..., or system under a lockout/tags-plus application. (4) Capable of being locked out. An energy-isolating... other means of attachment to which, or through which, a lock can be affixed. Other energy-isolating...

  11. 29 CFR 1915.80 - Scope, application, definitions, and effective dates.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... that provides an impediment (in addition to the energy-isolating device) to the release of energy or..., or system under a lockout/tags-plus application. (4) Capable of being locked out. An energy-isolating... other means of attachment to which, or through which, a lock can be affixed. Other energy-isolating...

  12. Framework for near-field-communication-based geo-localization and personalization for Android-based smartphones--application in hospital environments.

    PubMed

    Meng, Philipp; Fehre, Karsten; Rappelsberger, Andrea; Adlassnig, Klaus-Peter

    2014-01-01

    Various applications using near field communication (NFC) have been developed for the medical sector. As a method of short-range wireless contact-driven data transfer, NFC is a useful tool in medicine. It can be used to transfer data such as blood pressure, control adherence to medication, or transmit in vivo data. The first proposed general framework uses NFC as a mechanism for indoor geo-localization in hospitals. NFC geo-localization is economical compared to classical concepts using indoor GPS or WLAN triangulation, and the granularity of location retrieval can be defined at a tag level. Using this framework, we facilitate the development of medical applications that require exact indoor geo-localization. Multi-user Android systems are addressed in the second framework. Using private NFC tags, users are able to carry on their personal settings for enabled applications. This eliminates the need for multiple user accounts on common Android devices, improves usability, and eases technical administration. Based on the prototypes presented here, we show a novel concept of using NFC-enabled Android devices in hospital environments.

  13. Labeling milk along its production chain with DNA encapsulated in silica.

    PubMed

    Bloch, Madeleine S; Paunescu, Daniela; Stoessel, Philipp R; Mora, Carlos A; Stark, Wendelin J; Grass, Robert N

    2014-10-29

    The capability of tracing a food product along its production chain is important to ensure food safety and product authenticity. For this purpose and as an application example, recently developed Silica Particles with Encapsulated DNA (SPED) were added to milk at concentrations ranging from 0.1 to 100 ppb (μg per kg milk). Thereby the milk, as well as the milk-derived products yoghurt and cheese, could be uniquely labeled with a DNA tag. Procedures for the extraction of the DNA tags from the food matrixes were elaborated and allowed identification and quantification of previously marked products by quantitative polymerase chain reaction (qPCR) with detection limits below 1 ppb of added particles. The applicability of synthetic as well as naturally occurring DNA sequences was shown. The usage of approved food additives as DNA carrier (silica = E551) and the low cost of the technology (<0.1 USD per ton of milk labeled with 10 ppb of SPED) display the technical applicability of this food labeling technology.

  14. Beta-carbonic anhydrases play a role in fruiting body development and ascospore germination in the filamentous fungus Sordaria macrospora.

    PubMed

    Elleuche, Skander; Pöggeler, Stefanie

    2009-01-01

    Carbon dioxide (CO(2)) is among the most important gases for all organisms. Its reversible interconversion to bicarbonate (HCO(3) (-)) reaches equilibrium spontaneously, but slowly, and can be accelerated by a ubiquitous group of enzymes called carbonic anhydrases (CAs). These enzymes are grouped by their distinct structural features into alpha-, beta-, gamma-, delta- and zeta-classes. While physiological functions of mammalian, prokaryotic, plant and algal CAs have been extensively studied over the past years, the role of beta-CAs in yeasts and the human pathogen Cryptococcus neoformans has been elucidated only recently, and the function of CAs in multicellular filamentous ascomycetes is mostly unknown. To assess the role of CAs in the development of filamentous ascomycetes, the function of three genes, cas1, cas2 and cas3 (carbonic anhydrase of Sordaria) encoding beta-class carbonic anhydrases was characterized in the filamentous ascomycetous fungus Sordaria macrospora. Fluorescence microscopy was used to determine the localization of GFP- and DsRED-tagged CAs. While CAS1 and CAS3 are cytoplasmic enzymes, CAS2 is localized to the mitochondria. To assess the function of the three isoenzymes, we generated knock-out strains for all three cas genes (Deltacas1, Deltacas2, and Deltacas3) as well as all combinations of double mutants. No effect on vegetative growth, fruiting-body and ascospore development was seen in the single mutant strains lacking cas1 or cas3, while single mutant Deltacas2 was affected in vegetative growth, fruiting-body development and ascospore germination, and the double mutant strain Deltacas1/2 was completely sterile. Defects caused by the lack of cas2 could be partially complemented by elevated CO(2) levels or overexpression of cas1, cas3, or a non-mitochondrial cas2 variant. The results suggest that CAs are required for sexual reproduction in filamentous ascomycetes and that the multiplicity of isoforms results in redundancy of specific and non-specific functions.

  15. Cloning, expression in Pichia pastoris, and characterization of a thermostable GH5 mannan endo-1,4-beta-mannosidase from Aspergillus niger BK01.

    PubMed

    Do, Bien-Cuong; Dang, Thi-Thu; Berrin, Jean-Guy; Haltrich, Dietmar; To, Kim-Anh; Sigoillot, Jean-Claude; Yamabhai, Montarop

    2009-11-13

    Mannans are key components of lignocellulose present in the hemicellulosic fraction of plant primary cell walls. Mannan endo-1,4-beta-mannosidases (1,4-beta-D-mannanases) catalyze the random hydrolysis of beta-1,4-mannosidic linkages in the main chain of beta-mannans. Biodegradation of beta-mannans by the action of thermostable mannan endo-1,4-beta-mannosidase offers significant technical advantages in biotechnological industrial applications, i.e. delignification of kraft pulps or the pretreatment of lignocellulosic biomass rich in mannan for the production of second generation biofuels, as well as for applications in oil and gas well stimulation, extraction of vegetable oils and coffee beans, and the production of value-added products such as prebiotic manno-oligosaccharides (MOS). A gene encoding mannan endo-1,4-beta-mannosidase or 1,4-beta-D-mannan mannanohydrolase (E.C. 3.2.1.78), commonly termed beta-mannanase, from Aspergillus niger BK01, which belongs to glycosyl hydrolase family 5 (GH5), was cloned and successfully expressed heterologously (up to 243 microg of active recombinant protein per mL) in Pichia pastoris. The enzyme was secreted by P. pastoris and could be collected from the culture supernatant. The purified enzyme appeared glycosylated as a single band on SDS-PAGE with a molecular mass of approximately 53 kDa. The recombinant beta-mannanase is highly thermostable with a half-life time of approximately 56 h at 70 degrees C and pH 4.0. The optimal temperature (10-min assay) and pH value for activity are 80 degrees C and pH 4.5, respectively. The enzyme is not only active towards structurally different mannans but also exhibits low activity towards birchwood xylan. Apparent Km values of the enzyme for konjac glucomannan (low viscosity), locust bean gum galactomannan, carob galactomannan (low viscosity), and 1,4-beta-D-mannan (from carob) are 0.6 mg mL-1, 2.0 mg mL-1, 2.2 mg mL-1 and 1.5 mg mL-1, respectively, while the kcat values for these substrates are 215 s-1, 330 s-1, 292 s-1 and 148 s-1, respectively. Judged from the specificity constants kcat/Km, glucomannan is the preferred substrate of the A. niger beta -mannanase. Analysis by thin layer chromatography showed that the main product from enzymatic hydrolysis of locust bean gum is mannobiose, with only low amounts of mannotriose and higher manno-oligosaccharides formed. This study is the first report on the cloning and expression of a thermostable mannan endo-1,4-beta-mannosidase from A. niger in Pichia pastoris. The efficient expression and ease of purification will significantly decrease the production costs of this enzyme. Taking advantage of its acidic pH optimum and high thermostability, this recombinant beta-mannanase will be valuable in various biotechnological applications.

  16. In-Depth Lipidomic Analysis of Molecular Species of Triacylglycerides, Diacylglycerides, Glycerophospholipids, and Sphingolipids of Buttermilk by GC-MS/FID, HPLC-ELSD, and UPLC-QToF-MS

    PubMed Central

    Castro-Gómez, Pilar; Montero, Olimpio; Fontecha, Javier

    2017-01-01

    Buttermilk, a byproduct of butter manufacturing, has gained considerable attention due to its high concentration of polar lipids as phospho- and sphingolipids from the milk fat globule membrane (MFGM). These polar lipids (PLs) are essential components of all cellular membranes and exert a variety of indispensable metabolic, neurological, and intracellular signaling processes. Despite its importance, there are few research studies that report a comprehensive characterization of the lipid molecular species of MFGM that could contribute to a better understanding of their putative healthful activities. In this study, procedures such as pressurized liquid extraction of polar and nonpolar lipids and their fractionation by flash chromatography have been carried out. The obtained fractions were submitted to an exhaustive characterization from a lipidomic point of view. The characterization includes new data about the identification and quantification of triacylglycerides (TAG), diacylglycerides (DAG), and phospho- and sphingolipids using different chromatographic techniques. The fatty acid profile was comparable to that of the milk fat but with a highly diverse composition of fatty acids. Molecular species have also been determined by using ultra-high performance liquid chromatography/quadruple-time-of-flight mass spectrometry (UPLC/QToF-MS). The TAG (16:0/16:0/6:0) and TAG (16:0/16:0/8:0) were the predominant saturated TAG species and TAG (14:0/18:1/16:0) and TAG (16:0/16:0/18:1) presented the highest content of monounsaturated TAG species. Furthermore; over 30 molecular species of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylinositol (PI) could be identified within PL, with PC (16:0/18:1) being the most abundant species. Whereas C16:0 was found to be the preferred FA in TAGs, it was C18:1 in PLs. Several ganglioside species have also been characterized with d18:1 ceramide moiety and secondary acyl chains ranging from C20:0 to C26:1. This approach could broaden the applications of high-resolution mass spectrometry for a better understanding of the role of MFGM and its functionality. PMID:28287421

  17. The development and application of new crystallization method for tobacco mosaic virus coat protein.

    PubMed

    Li, Xiangyang; Song, Baoan; Hu, Deyu; Wang, Zhenchao; Zeng, Mengjiao; Yu, Dandan; Chen, Zhuo; Jin, Linhong; Yang, Song

    2012-11-21

    Although tobacco mosaic virus (TMV) coat protein (CP) has been isolated from virus particles and its crystals have grown in ammonium sulfate buffers for many years, to date, no one has reported on the crystallization of recombinant TMV-CP connecting peptides expressed in E. coli. In the present papers genetically engineered TMV-CP was expressed, into which hexahistidine (His) tags or glutathione-S-transferase (GST) tags were incorporated. Considering that GST-tags are long peptides and His-tags are short peptides, an attempt was made to grow crystals of TMV-CP cleaved GST-tags (WT-TMV-CP32) and TMV-CP incorporated His-tags (WT-His-TMV-CP12) simultaneously in ammonium sulfate buffers and commercial crystallization reagents. It was found that the 20S disk form of WT-TMV-CP32 and WT-His-TMV-CP12 did not form high resolution crystals by using various crystallization buffers and commercial crystallization reagents. Subsequently, a new experimental method was adopted in which a range of truncated TMV-CP was constructed by removing several amino acids from the N- or the C-terminal, and high resolution crystals were grown in ammonium sulfate buffers and commercial crystallization reagents. The new crystallization method was developed and 3.0 Å resolution macromolecular crystal was thereby obtained by removing four amino acids at the C-terminal of His-TMV-CP and connecting six His-tags at the N-terminal of His-TMV-CP (TR-His-TMV-CP19). The Four-layer aggregate disk structure of TR-His-TMV-CP19 was solved. This phenomenon showed that peptides at the C-terminus hindered the growth of high resolution crystals and the peptides interactions at the N-terminus were attributed to the quality of TMV-CP crystals. A 3.0 Å resolution macromolecular crystal of TR-His-TMV-CP19 was obtained and the corresponding structure was solved by removing four amino acids at the C-terminus of TMV-CP and connecting His-tags at the N-terminus of TMV-CP. It indicated that short peptides influenced the resolution of TMV-CP crystals.

  18. Raft and floating radio frequency identification (RFID) antenna systems for detecting and estimating abundance of PIT-tagged fish in rivers

    USGS Publications Warehouse

    Fetherman, Eric R.; Avila, Brian W.; Winkelman, Dana L.

    2016-01-01

    Portable radio frequency identification (RFID) PIT tag antenna systems are increasingly being used in studies examining aquatic animal movement, survival, and habitat use, and their design flexibility permits application in a wide variety of settings. We describe the construction, use, and performance of two portable floating RFID PIT tag antenna systems designed to detect fish that were unavailable for recapture using stationary antennas or electrofishing. A raft antenna system was designed to detect and locate PIT-tagged fish in relatively long (i.e., ≥10 km) river reaches, and consisted of two antennas: (1) a horizontal antenna (4 × 1.2 m) installed on the bottom of the raft and used to detect fish in shallower river reaches (<1 m), and (2) a vertical antenna (2.7 × 1.2 m) for detecting fish in deeper pools (≥1 m). Detection distances of the horizontal antenna were between 0.7 and 1.0 m, and detection probability was 0.32 ± 0.02 (mean ± SE) in a field test using rocks marked with 32-mm PIT tags. Detection probability of PIT-tagged fish in the Cache la Poudre River, Colorado, using the raft antenna system, which covered 21% of the wetted area, was 0.14 ± 0.14. A shore-deployed floating antenna (14.6 × 0.6 m), which covered 100% of the wetted area, was designed for use by two operators for detecting and locating PIT-tagged fish in shorter (i.e., <2 km) river reaches. Detection distances of the shore-deployed floating antenna were between 0.7 and 0.8 m, and detection probabilities during field deployment in the St. Vrain River exceeded 0.52. The shore-deployed floating antenna was also used to estimate abundance of PIT-tagged fish. Results suggest that the shore-deployed floating antenna could be used as an alternative to estimating abundance using traditional sampling methods such as electrofishing.

  19. Effect of geometrical configuration of radioactive sources on radiation intensity in beta-voltaic nuclear battery system: A preliminary result

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Basar, Khairul, E-mail: khbasar@fi.itb.ac.id; Riupassa, Robi D., E-mail: khbasar@fi.itb.ac.id; Bachtiar, Reza, E-mail: khbasar@fi.itb.ac.id

    2014-01-01

    It is known that one main problem in the application of beta-voltaic nuclear battery system is its low efficiency. The efficiency of the beta-voltaic nuclear battery system mainly depends on three aspects: source of radioactive radiation, interface between materials in the system and process of converting electron-hole pair to electric current in the semiconductor material. In this work, we show the effect of geometrical configuration of radioactive sources on radiation intensity of beta-voltaic nuclear battery system.

  20. Regulation of the catalytic behavior of pullulanases chelated onto nickel (II)-modified magnetic nanoparticles.

    PubMed

    Wang, Jianfeng; Liu, Zhongmei; Zhou, Zhemin

    2017-06-01

    Chelating of pullulanases onto nickel (II)-modified magnetic nanoparticles results in one-step purification and immobilization of pullulanase, and facilitates the commercial application of pullulanase in industrial scale. To improve the catalytic behavior, especially the operational stability, of the nanocatalyst in consecutive batch reactions, we prepared various iminodiacetic acid-modified magnetic nanoparticles differed in surface polarity and spacer length, on which the His6-tagged pullulanases were chelated via nickel ions, and then studied the correlation between the MNPs surface property and the corresponding catalyst behavior. When pullulanases were chelated onto the surface-modified MNPs, the thermostability of all pullulanase derivatives were lower than that of free counterpart, being not relevant to the protein orientation guided by the locality of the His6-tag, but related to the MNPs basal surface polarity and the grafted spacer length. After chelating of pullulanases onto MNPs, there were changes observed in the pH-activity profile and the apparent Michaelis constant toward pullulan. The changing tendencies were mainly dependent on the His6-tagged pullulanase orientation, and the changing extents were tuned by the spacer length. The reusability of pullulanase immobilized by N-terminal His6-tag was higher than that of pullulanase immobilized by C-terminal His6-tag. Moreover, the reusability of the immobilized pullulanase tested increased till grafting polyether amine-400 as spacer-arm, therefore the N-terminal His6-tagged pullulanase chelating MNPs grafted polyether amine-400 gave the best reusability, which retained 60% of initial activity after 18 consecutive cycles with a total reaction time of 9h. Additionally, the correlation analysis of the catalyst behaviors indicated that the reusability was independent from other catalytic properties such as thermostability and substrate affinity. All the results revealed that the catalyst behavior can be mainly controlled by the His6-tagged pullulanase orientation than by the MNPs surface property which can tune the catalyst function. Copyright © 2017. Published by Elsevier Inc.

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