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Sample records for neutral activation analysis

  1. Evaluation and analysis of dengue virus enhancing and neutralizing activities using simple high-throughput assays.

    PubMed

    Li, Xiao-Quan; Chen, Jing; Huang, Yan-Fen; Ding, Xi-Xia; Liu, Li-Dong; Qiu, Li-Wen; Pan, Yu-Xian; Deng, Yong-Qiang; Hu, Dong-Mei; Di, Biao; Qin, Cheng-Feng; Che, Xiao-Yan

    2013-07-01

    The risk of antibody-dependent enhancement (ADE) of dengue virus (DENV) infection is a major obstacle for the development of dengue vaccine candidates. Here, we described a novel approach for assessment of ADE by measuring DENV nonstructural protein 1 (NS1) production in culture supernatants with Fcγ receptor-expressing K562 cells in ELISA format (ELISA-ADE). Enhancing activities quantified by measurement of kinetics of NS1 production were in a good agreement with the results of the virus titration assay. In conjunction with the previously established enzyme-linked immunospot-based micro-neutralization test (ELISPOT-MNT) in 96-well format, the observable dose-response profiles of enhancing and neutralizing activities against all four DENV serotypes were produced with two flaviviral envelope cross-reactive monoclonal antibodies and four primary DENV-1-infected human sera. The simple high-throughput ELISA-ADE assay offers advantages for quantitative measurement of infection enhancement that can potentially be applied to large-scale seroepidemiological studies of DENV infection and vaccination.

  2. Analysis of Non-Enzymatically Glycated Peptides: Neutral-Loss Triggered MS3 Versus Multi-Stage Activation Tandem Mass Spectrometry

    SciTech Connect

    Zhang, Qibin; Petyuk, Vladislav A.; Schepmoes, Athena A.; Orton, Daniel J.; Monroe, Matthew E.; Yang, Feng; Smith, Richard D.; Metz, Thomas O.

    2008-10-15

    Non-enzymatic glycation of tissue proteins has important implications in the development of complications of diabetes mellitus. While electron transfer dissociation (ETD) has been shown to outperform collision-induced dissociation (CID) in sequencing glycated peptides by tandem mass spectrometry, ETD instrumentation is not yet available in all laboratories. In this study, we evaluated different advanced CID techniques (i.e., neutral-loss triggered MS3 and multi-stage activation) during LC-MSn analyses of Amadori-modified peptides enriched from human serum glycated in vitro. During neutral-loss triggered MS3 experiments, MS3 scans triggered by neutral-losses of 3 H2O or 3 H2O + HCHO produced similar results in terms of glycated peptide identifications. However, neutral losses of 3 H2O resulted in significantly more glycated peptide identifications during multi-stage activation experiments. Overall, the multi-stage activation approach produced more glycated peptide identifications, while the neutral-loss triggered MS3 approach resulted in much higher specificity. Both techniques offer a viable alternative to ETD for identifying glycated peptides when that method is unavailable.

  3. Durability of bactericidal activity in electrolyzed neutral water by storage.

    PubMed

    Nagamatsu, Yuki; Chen, Kerr-Kong; Tajima, Kiyoshi; Kakigawa, Hiroshi; Kozono, Yoshio

    2002-06-01

    Electrolyzed strong and weak acid waters have been widely used for sterilization in clinical dentistry because of their excellent bactericidal activities. Electrolyzed neutral water was recently developed with a new concept of long-term good durability in addition to the excellent bactericidal activity similar to acid waters. The present study, evaluated the storage life of this water compared with the acid waters in terms of the changes in pH, oxidation-reduction potential (ORP), residual chlorine and bactericidal activity under several conditions using Staphylococcus aureus 209P. The strong acid water showed a rapid deterioration of its bactericidal activity. The weak acid and neutral waters exhibited excellent durability. Although all the bacteria were annihilated by the contact with the waters even stored for 40 days in the uncapped bottle, the neutral water was superior in further long-term duration.

  4. Multivariate analysis of human immunodeficiency virus type 1 neutralization data.

    PubMed Central

    Nyambi, P N; Nkengasong, J; Lewi, P; Andries, K; Janssens, W; Fransen, K; Heyndrickx, L; Piot, P; van der Groen, G

    1996-01-01

    We report on the use of spectral map analysis of the inter- and intraclade neutralization data of 14 sera of human immunodeficiency virus type 1 (HIV-1)-infected individuals and 16 primary isolates, representing genetic clades A to H in group M and group O. This multivariate analysis has been used previously to study the interaction between drugs and receptors and between viruses and antiviral compounds. The analysis reveals the existence of neutralization clusters, not correlated with the known genetic clades. The structural factors that have been identified may correlate with the most important neutralization epitopes. Three key primary HIV-1 isolates, which allow discrimination of sera that are likely or unlikely to neutralize primary isolates from most of the genetic clades, were identified. Our method of analysis will facilitate the evaluation as well as the design of suitable HIV-1 vaccines, which induce high-titer interclade cross-neutralizing antibodies. PMID:8709250

  5. Thermochemical Analysis of Neutralization Reactions: An Introductory Discovery Experiment

    ERIC Educational Resources Information Center

    Mills, Kenneth V.; Gullmette, Louise W.

    2007-01-01

    The article describes a new discovery experiment that uses thermodynamical analysis to study neutralization reactions based on neutralization of citric acid. The experiment would be able to reinforce students' understanding of stoichiometry and allow for the discovery of basic concepts of thermochemistry.

  6. Seismic active control by neutral networks

    SciTech Connect

    Tang, Yu

    1995-12-31

    A study on the application of artificial neural networks (ANNs) to active structural control under seismic loads is carried out. The structure considered is a single-degree-of-freedom (SDF) system with an active bracing device. The control force is computed by a trained neural network. The feedforward neural network architecture and an adaptive backpropagation training algorithm is used in the study. The neural net is trained to reproduce the function that represents the response-excitation relationship of the SDF system under seismic loads. The input-output training patterns are generated randomly. In the backpropagation training algorithm, the learning rate is determined by ensuring the decrease of the error function at each epoch. The computer program implemented is validated by solving the classification of the XOR problem. Then, the trained ANN is used to compute the control force according to the control strategy. If the control force exceeds the actuator`s capacity limit, it is set equal to that limit. The concept of the control strategy employed herein is to apply the control force at every time step to cancel the system velocity induced at the preceding time step so that the gradual rhythmic buildup of the response is destroyed. The ground motions considered in the numerical example are the 1940 El Centro earthquake and the 1979 Imperial Valley earthquake in California. The system responses with and without the control are calculated and compared. The feasibility and potential of applying ANNs to seismic active control is asserted by the promising results obtained from the numerical examples studied.

  7. STATISTICAL ANALYSIS OF THE HEAVY NEUTRAL ATOMS MEASURED BY IBEX

    SciTech Connect

    Park, Jeewoo; Kucharek, Harald; Möbius, Eberhard; Galli, André; Livadiotis, George; Fuselier, Steve A.; McComas, David J.

    2015-10-15

    We investigate the directional distribution of heavy neutral atoms in the heliosphere by using heavy neutral maps generated with the IBEX-Lo instrument over three years from 2009 to 2011. The interstellar neutral (ISN) O and Ne gas flow was found in the first-year heavy neutral map at 601 keV and its flow direction and temperature were studied. However, due to the low counting statistics, researchers have not treated the full sky maps in detail. The main goal of this study is to evaluate the statistical significance of each pixel in the heavy neutral maps to get a better understanding of the directional distribution of heavy neutral atoms in the heliosphere. Here, we examine three statistical analysis methods: the signal-to-noise filter, the confidence limit method, and the cluster analysis method. These methods allow us to exclude background from areas where the heavy neutral signal is statistically significant. These methods also allow the consistent detection of heavy neutral atom structures. The main emission feature expands toward lower longitude and higher latitude from the observational peak of the ISN O and Ne gas flow. We call this emission the extended tail. It may be an imprint of the secondary oxygen atoms generated by charge exchange between ISN hydrogen atoms and oxygen ions in the outer heliosheath.

  8. Launch Deployment Assembly Extravehicular Activity Neutral Buoyancy Development Test Report

    NASA Technical Reports Server (NTRS)

    Loughead, T.

    1996-01-01

    This test evaluated the Launch Deployment Assembly (LDA) design for Extravehicular Activity (EVA) work sites (setup, igress, egress), reach and visual access, and translation required for cargo item removal. As part of the LDA design, this document describes the method and results of the LDA EVA Neutral Buoyancy Development Test to ensure that the LDA hardware support the deployment of the cargo items from the pallet. This document includes the test objectives, flight and mockup hardware description, descriptions of procedures and data collection used in the testing, and the results of the development test at the National Aeronautics and Space Administrations (NASA) Marshall Space Flight Center (MSFC) Neutral Buoyancy Simulator (NBS).

  9. Lunar Neutral Exposphere Properties from Pickup Ion Analysis

    NASA Technical Reports Server (NTRS)

    Hartle, R. E.; Sarantos, M.; Killen, R.; Sittler, E. C. Jr.; Halekas, J.; Yokota, S.; Saito, Y.

    2009-01-01

    , yielding the necessary ion velocity spectra needed to further analysis of parent neutral exosphere properties.

  10. Analysis of neutral hydrogenic emission spectra in a tokamak

    NASA Astrophysics Data System (ADS)

    Ko, J.; Chung, J.; Jaspers, R. J. E.

    2015-10-01

    Balmer-α radiation by the excitation of thermal and fast neutral hydrogenic particles has been investigated in a magnetically confined fusion device, or tokamak, from the Korea Superconducting Tokamak Advanced Research (KSTAR). From the diagnostic point of view, the emission from thermal neutrals is associated with passive spectroscopy and that from energetic neutrals that are usually injected from the outside of the tokamak to the active spectroscopy. The passive spectroscopic measurement for the thermal Balmer-α emission from deuterium and hydrogen estimates the relative concentration of hydrogen in a deuterium-fueled plasma and therefore, makes a useful tool to monitor the vacuum wall condition. The ratio of hydrogen to deuterium obtained from this measurement qualitatively correlates with the energy confinement of the plasma. The Doppler-shifted Balmer-α components from the fast neutrals features the spectrum of the motional Stark effect (MSE) which is an essential principle for the measurement of the magnetic pitch angle profile. Characterization of this active MSE spectra, especially with multiple neutral beam lines crossing along the observation line of sight, has been done for the guideline of the multi-ion-source heating beam operation and for the optimization of the narrow bandpass filters that are required for the polarimeter-based MSE diagnostic system under construction at KSTAR.

  11. Analysis of Titan's neutral upper atmosphere from Cassini Ion Neutral Mass Spectrometer measurements

    NASA Astrophysics Data System (ADS)

    Cui, J.; Yelle, R. V.; Vuitton, V.; Waite, J. H.; Kasprzak, W. T.; Gell, D. A.; Niemann, H. B.; Müller-Wodarg, I. C. F.; Borggren, N.; Fletcher, G. G.; Patrick, E. L.; Raaen, E.; Magee, B. A.

    2009-04-01

    In this paper we present an in-depth study of the distributions of various neutral species in Titan's upper atmosphere, between 950 and 1500 km for abundant species (N 2, CH 4, H 2) and between 950 and 1200 km for other minor species. Our analysis is based on a large sample of Cassini/INMS (Ion Neutral Mass Spectrometer) measurements in the CSN (Closed Source Neutral) mode, obtained during 15 close flybys of Titan. To untangle the overlapping cracking patterns, we adopt Singular Value Decomposition (SVD) to determine simultaneously the densities of different species. Except for N 2, CH 4, H 2 and 40Ar (as well as their isotopes), all species present density enhancements measured during the outbound legs. This can be interpreted as a result of wall effects, which could be either adsorption/desorption of these molecules or heterogeneous surface chemistry of the associated radicals on the chamber walls. In this paper, we provide both direct inbound measurements assuming ram pressure enhancement only and abundances corrected for wall adsorption/desorption based on a simple model to reproduce the observed time behavior. Among all minor species of photochemical interest, we have firm detections of C 2H 2, C 2H 4, C 2H 6, CH 3C 2H, C 4H 2, C 6H 6, CH 3CN, HC 3N, C 2N 2 and NH 3 in Titan's upper atmosphere. Upper limits are given for other minor species. The globally averaged distributions of N 2, CH 4 and H 2 are each modeled with the diffusion approximation. The N 2 profile suggests an average thermospheric temperature of 151 K. The CH 4 and H 2 profiles constrain their fluxes to be 2.6×10 cms and 1.1×10 cms, referred to Titan's surface. Both fluxes are significantly higher than the Jeans escape values. The INMS data also suggest horizontal/diurnal variations of temperature and neutral gas distribution in Titan's thermosphere. The equatorial region, the ramside, as well as the nightside hemisphere of Titan appear to be warmer and present some evidence for the depletion

  12. Neutral density filters with Risley prisms: analysis and design.

    PubMed

    Duma, Virgil-Florin; Nicolov, Mirela

    2009-05-10

    We achieve the analysis and design of optical attenuators with double-prism neutral density filters. A comparative study is performed on three possible device configurations; only two are presented in the literature but without their design calculus. The characteristic parameters of this optical attenuator with Risley translating prisms for each of the three setups are defined and their analytical expressions are derived: adjustment scale (attenuation range) and interval, minimum transmission coefficient and sensitivity. The setups are compared to select the optimal device, and, from this study, the best solution for double-prism neutral density filters, both from a mechanical and an optical point of view, is determined with two identical, symmetrically movable, no mechanical contact prisms. The design calculus of this optimal device is developed in essential steps. The parameters of the prisms, particularly their angles, are studied to improve the design, and we demonstrate the maximum attenuation range that this type of attenuator can provide.

  13. Identification of neutral and acidic deoxyribonuclease activities in Tetrahymena thermophila life stages.

    PubMed

    Aslan, Erhan; Arslanyolu, Muhittin

    2015-04-01

    Deoxyribonucleases (DNases) play a major role in apoptotic DNA fragmentation/degradation, and apoptotic-like DNA degradation is also observed during conjugation of the ciliate Tetrahymena thermophila; however, the characteristics of neutral and acidic DNases are still undefined in its life stages. Here, we report the biochemical characterization of DNase activities displayed in three different Tetrahymena life stages in a comparative manner. Maximum DNase activity of Tetrahymena was observed under acidic conditions, indicating that Tetrahymena has strong DNase II-like activities. Zymography revealed that Tetrahymena has at least five distinct DNase activity bands at 28, 32, 33.8, 35.5, and 69-kDa, and that the activities at 32 and 33.8-kDa were also secreted into starvation buffer. Cofactor analysis demonstrated that Mg(2+) exerted inhibitory effects on neutral DNase activities. Unexpectedly, Mg(2+) and Ca(2+) had favorable effects on acidic DNase activities. The DNase activity profile of conjugating Tetrahymena cells revealed that the 32 and 33.8-kDa activities at pH 5.0 increased from 14 to 18 h of conjugation, corresponding to the final resorption of the old macronucleus by lysosomal enzymes during programmed nuclear death (PND). Overall, we found that Tetrahymena DNases exhibit different biochemical properties and a possible involvement of DNase II-like activities in PND.

  14. Magnitudes of muscle activation of spine stabilizers, gluteals, and hamstrings during supine bridge to neutral position.

    PubMed

    Youdas, James W; Hartman, James P; Murphy, Brooke A; Rundle, Ashley M; Ugorowski, Jenna M; Hollman, John H

    2015-01-01

    The aim of this study was to compare the magnitude of selective core muscle activation during supine bridging to neutral exercises (three on a stable and three on an unstable surface). Surface EMG analysis was performed on the lumbar multifidus, gluteus medius, gluteus maximus, and hamstrings from 13 male and 13 female subjects. Lumbar multifidus recruitment was not influenced by exercise or condition and ranged between 29.2 and 35.9% of maximum voluntary isometric contraction (MVIC). Peak gluteus medius activation (42.0% MVIC) occurred in unstable single-leg bridge. Maximum recruitment of gluteus maximus (32.6% MVIC) appeared during stable single-leg bridge. Peak hamstring activation (59.6% MVIC) occurred during stable double-leg hamstring curl. Regardless of condition, hamstrings demonstrated high (51.9-59.6% MVIC) muscle recruitment during double-leg hamstring curls compared with the single-leg bridge or double-leg bridge. Various supine bridging to neutral exercises activated the hamstrings at levels conducive to strengthening, whereas recruitment of lumbar multifidus, gluteus medius, and gluteus maximus promoted endurance training. Clinically, we were unable to conclude the unstable support surface was preferable to the stable surface for boosting muscle recruitment of spine stabilizers, gluteals, and hamstring muscles during supine bridge to neutral position. PMID:25671354

  15. Mechanical Activation-Assisted Reductive Leaching of Cadmium from Zinc Neutral Leaching Residue Using Sulfur Dioxide

    NASA Astrophysics Data System (ADS)

    Zhang, Chun; Min, Xiaobo; Chai, Liyuan; Zhang, Jianqiang; Wang, Mi

    2015-12-01

    In this work, zinc neutral leaching residue was mechanically activated by ball-milling. The subsequent leaching behavior and kinetics of cadmium extraction in a mixed SO2-H2SO4 system were studied. Changes in the crystalline phase, lattice distortion, particle size and morphology, which were induced by mechanical activation, were also investigated. The activated samples showed different physicochemical characteristics, and cadmium extraction was found to be easier than for the un-activated samples. Under the same conditions, mechanical activation contributed to higher cadmium leaching. The cadmium extraction kinetics at 75-95°C was found to fit the shrinking core model. The raw neutral leaching residue, and the samples activated for 60 min and 120 min had a calculated activation energy of 65.02 kJ/mol, 59.45 kJ/mol and 53.46 kJ/mol, respectively. The leaching residue was characterized by ICP, XRD and SEM analysis. According to XRD analysis, the main phases in the residue were lead sulfate (PbSO4), zinc sulfide (ZnS) and cadmium sulfide (CdS).

  16. Activation analysis

    SciTech Connect

    Alfassi, Z.B. . Dept. of Nuclear Engineering)

    1990-01-01

    This volume contains 16 chapters on the application of activation analysis in the fields of life sciences, biological materials, coal and its effluents, environmental samples, archaeology, material science, and forensics. Each chapter is processed separately for the data base.

  17. Au@Ag Heterogeneous Nanorods as Nanozyme Interfaces with Peroxidase-Like Activity and Their Application for One-Pot Analysis of Glucose at Nearly Neutral pH.

    PubMed

    Han, Lei; Li, Cuncheng; Zhang, Tao; Lang, Qiaolin; Liu, Aihua

    2015-07-01

    As substitutes for natural peroxidases, most nanomaterial-based enzyme mimetics (nanozymes) have unique properties such as high stability, low-cost, large surface area, and high catalytic activity. However, they usually work in acidic conditions and thus impede their real applications. In this work, by modulating the nanostructure, composition, and surface property of the bimetallic materials, the positively charged poly(diallyldimethylammonium)-stabilized Au@Ag heterogeneous nanorods (NRs) were developed as synergistic peroxidase-like interfaces, which exhibited high activity over a wide pH range (pH 4.0-6.5) using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) as the chromogenic substrate. At pH 6.5, the peroxidase-like activity for the Au@Ag heterogeneous NRs was stable and optimal within 20-40 °C. Moreover, the Au@Ag heterogeneous NRs showed excellent temperature stability and long-term storage stability. Given these characters, the detection of H2O2 at pH 6.5 was proposed on the basis of the Au@Ag heterogeneous NRs catalyzing the colorimetric reaction of H2O2 and ABTS, where the oxidized ABTS showed a typical absorption peak at 414 nm. The absorbance at 414 nm was linear with H2O2 concentration from 0.01 to 10 mM. Further, considering that Au@Ag heterogeneous NRs and glucose oxidase (GOx) have similar optimal pH for catalytic activities, a novel one-pot method for the detection of glucose was developed by the coupled catalytic reaction using GOx, Au@Ag heterogeneous NRs, and ABTS at nearly neutral pH (pH 6.5) and 37 °C. This proposed method had simple and rapid processes, wide linear range (0.05-20 mM), and reliability for the successful analysis of real samples. On the basis of these attractive and unique characteristics, Au@Ag heterogeneous NRs can become promising substitutes for peroxidase in analytical chemistry and environmental science.

  18. Au@Ag Heterogeneous Nanorods as Nanozyme Interfaces with Peroxidase-Like Activity and Their Application for One-Pot Analysis of Glucose at Nearly Neutral pH.

    PubMed

    Han, Lei; Li, Cuncheng; Zhang, Tao; Lang, Qiaolin; Liu, Aihua

    2015-07-01

    As substitutes for natural peroxidases, most nanomaterial-based enzyme mimetics (nanozymes) have unique properties such as high stability, low-cost, large surface area, and high catalytic activity. However, they usually work in acidic conditions and thus impede their real applications. In this work, by modulating the nanostructure, composition, and surface property of the bimetallic materials, the positively charged poly(diallyldimethylammonium)-stabilized Au@Ag heterogeneous nanorods (NRs) were developed as synergistic peroxidase-like interfaces, which exhibited high activity over a wide pH range (pH 4.0-6.5) using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) as the chromogenic substrate. At pH 6.5, the peroxidase-like activity for the Au@Ag heterogeneous NRs was stable and optimal within 20-40 °C. Moreover, the Au@Ag heterogeneous NRs showed excellent temperature stability and long-term storage stability. Given these characters, the detection of H2O2 at pH 6.5 was proposed on the basis of the Au@Ag heterogeneous NRs catalyzing the colorimetric reaction of H2O2 and ABTS, where the oxidized ABTS showed a typical absorption peak at 414 nm. The absorbance at 414 nm was linear with H2O2 concentration from 0.01 to 10 mM. Further, considering that Au@Ag heterogeneous NRs and glucose oxidase (GOx) have similar optimal pH for catalytic activities, a novel one-pot method for the detection of glucose was developed by the coupled catalytic reaction using GOx, Au@Ag heterogeneous NRs, and ABTS at nearly neutral pH (pH 6.5) and 37 °C. This proposed method had simple and rapid processes, wide linear range (0.05-20 mM), and reliability for the successful analysis of real samples. On the basis of these attractive and unique characteristics, Au@Ag heterogeneous NRs can become promising substitutes for peroxidase in analytical chemistry and environmental science. PMID:26076372

  19. Comprehensive Cross-Clade Neutralization Analysis of a Panel of Anti-Human Immunodeficiency Virus Type 1 Monoclonal Antibodies

    PubMed Central

    Binley, James M.; Wrin, Terri; Korber, Bette; Zwick, Michael B.; Wang, Meng; Chappey, Colombe; Stiegler, Gabriela; Kunert, Renate; Zolla-Pazner, Susan; Katinger, Hermann; Petropoulos, Christos J.; Burton, Dennis R.

    2004-01-01

    Broadly neutralizing monoclonal antibodies (MAbs) are potentially important tools in human immunodeficiency virus type 1 (HIV-1) vaccine design. A few rare MAbs have been intensively studied, but we still have a limited appreciation of their neutralization breadth. Using a pseudovirus assay, we evaluated MAbs from clade B-infected donors and a clade B HIV+ plasma against 93 viruses from diverse backgrounds. Anti-gp120 MAbs exhibited greater activity against clade B than non-B viruses, whereas anti-gp41 MAbs exhibited broad interclade activity. Unexpectedly, MAb 4E10 (directed against the C terminus of the gp41 ectodomain) neutralized all 90 viruses with moderate potency. MAb 2F5 (directed against an epitope adjacent to that of 4E10) neutralized 67% of isolates, but none from clade C. Anti-gp120 MAb b12 (directed against an epitope overlapping the CD4 binding site) neutralized 50% of viruses, including some from almost every clade. 2G12 (directed against a high-mannose epitope on gp120) neutralized 41% of the viruses, but none from clades C or E. MAbs to the gp120 V3 loop, including 447-52D, neutralized a subset of clade B viruses (up to 45%) but infrequently neutralized other clades (≤7%). MAbs b6 (directed against the CD4 binding site) and X5 (directed against a CD4-induced epitope of gp120) neutralized only sensitive primary clade B viruses. The HIV+ plasma neutralized 70% of the viruses, including some from all major clades. Further analysis revealed five neutralizing immunotypes that were somewhat associated with clades. As well as the significance for vaccine design, our data have implications for passive-immunization studies in countries where clade C viruses are common, given that only MAbs b12 and 4E10 were effective against viruses from this clade. PMID:15542675

  20. Computational analysis of anti-HIV-1 antibody neutralization panel data to identify potential functional epitope residues.

    PubMed

    West, Anthony P; Scharf, Louise; Horwitz, Joshua; Klein, Florian; Nussenzweig, Michel C; Bjorkman, Pamela J

    2013-06-25

    Advances in single-cell antibody cloning methods have led to the identification of a variety of broadly neutralizing anti-HIV-1 antibodies. We developed a computational tool (Antibody Database) to help identify critical residues on the HIV-1 envelope protein whose natural variation affects antibody activity. Our simplifying assumption was that, for a given antibody, a significant portion of the dispersion of neutralization activity across a panel of HIV-1 strains is due to the amino acid identity or glycosylation state at a small number of specific sites, each acting independently. A model of an antibody's neutralization IC50 was developed in which each site contributes a term to the logarithm of the modeled IC50. The analysis program attempts to determine the set of rules that minimizes the sum of the residuals between observed and modeled IC50 values. The predictive quality of the identified rules may be assessed in part by whether there is support for rules within individual viral clades. As a test case, we analyzed antibody 8ANC195, an anti-glycoprotein gp120 antibody of unknown specificity. The model for this antibody indicated that several glycosylation sites were critical for neutralization. We evaluated this prediction by measuring neutralization potencies of 8ANC195 against HIV-1 in vitro and in an antibody therapy experiment in humanized mice. These experiments confirmed that 8ANC195 represents a distinct class of glycan-dependent anti-HIV-1 antibody and validated the utility of computational analysis of neutralization panel data.

  1. JET divertor diagnostic upgrade for neutral gas analysis.

    PubMed

    Kruezi, Uron; Sergienko, G; Morgan, P D; Matthews, G F; Brezinsek, S; Vartanian, S

    2012-10-01

    With installation of the ITER-like wall in JET a major diagnostic upgrade to measure the neutral gas pressure and composition in the sub-divertor region has been completed, to characterise retention and outgassing of the new metallic first wall. The upgrade includes two new magnetically shielded systems consisting of sensitive capacitance manometers and residual gas analysers, both capable of providing data during plasma operation. These enable absolute pressure and gas composition measurements (pressure range: 10(-5)-10(-1) mbar, mass range: 1-200 amu, respectively) and have been used to characterise the neutral gas behaviour under various plasma conditions.

  2. An analysis of bipropellant neutralization for spacecraft refueling operations

    NASA Technical Reports Server (NTRS)

    Kauffman, David

    1987-01-01

    Refueling of satellites on orbit with storable propellants will involve venting part or all of the pressurant gas from the propellant tanks. This gas will be saturated with propellant vapor, and it may also have significant amounts of entrained fine droplets of propellant. The two most commonly used bipropellants, monomethyl hydrazine (MMH) and nitrogen tetroxide (N2O4), are highly reactive and toxic. Various possible ways of neutralizing the vented propellants are examined. The amount of propellant vented in a typical refueling operation is shown to be in the range of 0.2 to 5% of the tank capacity. Four potential neutralization schemes are examined: chemical decomposition, chemical reaction, condensation and adsorption. Chemical decomposition to essentially inert materials is thermodynamically feasible for both MMH and N2O4. It would be the simplest and easiest neutralization method to implement. Chemical decomposition would require more complex control. Condensation would require a refrigeration system and a very efficent phase separator. Adsorption is likely to be much heavier. A preliminary assessment of the four neutralization shemes is presented, along with suggested research and development plans.

  3. Values in the Net Neutrality Debate: Applying Content Analysis to Testimonies from Public Hearings

    ERIC Educational Resources Information Center

    Cheng, An-Shou

    2012-01-01

    The Net neutrality debate is an important telecommunications policy issue that closely tied to technological innovation, economic development, and information access. Existing studies on Net neutrality have focused primarily on technological requirements, economic analysis, and regulatory justifications. Since values, technology, and policy are…

  4. Nonlinear analysis of non-neutral plasmas. Report for 1 October 1986-30 September 1987

    SciTech Connect

    Ayres, V.M.

    1987-09-30

    The purpose of this investigation is to incorporate nonlinear effects into the analysis of non-neutral plasma beams in order to accurately estimate their performance in realistic situations. Intense non-neutral plasma beams have important applications for DEW, SDI, high power microwave/millimeter wave production, and particle-beam guidance systems.

  5. Neural Activation Underlying Cognitive Control in the Context of Neutral and Affectively Charged Pictures in Children

    ERIC Educational Resources Information Center

    Lamm, Connie; White, Lauren K.; McDermott, Jennifer Martin; Fox, Nathan A.

    2012-01-01

    The neural correlates of cognitive control for typically developing 9-year-old children were examined using dense-array ERPs and estimates of cortical activation (LORETA) during a go/no-go task with two conditions: a neutral picture condition and an affectively charged picture condition. Activation was estimated for the entire cortex after which…

  6. Heavy Neutral Beam Probe for Edge Plasma Analysis in Tokamaks

    SciTech Connect

    Castracane, J.

    2001-01-04

    The Heavy Neutral Beam Probe (HNBP) developed initially with DOE funding under the Small Business Innovation Research (SBIR) program was installed on the Tokamak de Varennes (TdeV) at the CCFM. This diagnostic was designed to perform fundamental measurements of edge plasma properties. The hardware was capable of measuring electron density and potential profiles with high spatial and temporal resolution. Fluctuation spectra for these parameters were obtained with HNBP for transport studies.

  7. Preclinical evaluation of three polyspecific antivenoms against the venom of Echis ocellatus: Neutralization of toxic activities and antivenomics.

    PubMed

    Calvete, Juan J; Arias, Ana Silvia; Rodríguez, Yania; Quesada-Bernat, Sarai; Sánchez, Laura V; Chippaux, Jean Philippe; Pla, Davinia; Gutiérrez, José María

    2016-09-01

    Snakebite envenoming has a heavy burden in the public health in sub-Saharan Africa. The viperid species Echis ocellatus (carpet viper or saw-scaled viper) is the medically most important snake in the savannahs of western sub-Saharan Africa. Several antivenoms are being distributed and used in this region for the treatment of envenomings by E. ocellatus, but the preclinical efficacy of some of these antivenoms has not been assessed. The present study evaluated the preclinical efficacy against E. ocellatus venom of three polyspecific antivenoms: (a) Snake Venom Antiserum (Pan Africa), manufactured by Premium Serums and Vaccines (India); (b) Snake Venom Antiserum (Africa), manufactured by VINS Bioproducts (India); and (c) Antivipmyn(®) Africa, manufactured by Instituto Bioclon (Mexico). Antivenomics analysis revealed the ability of the three antivenoms to immunocapture the majority of components of the venoms of E. ocellatus from Cameroon, Nigeria and Mali, although their maximal immunocapturing capability varied. Bioclon and Premium Serums antivenoms were effective in the neutralization of lethal, hemorrhagic and in vitro coagulant activities of the venom of E. ocellatus from Cameroon, albeit with different potencies. VINS antivenom neutralized hemorrhagic activity of this venom, but failed to neutralize lethality at the highest antivenom dose tested, and had a low neutralizing efficacy against in vitro coagulant effect.

  8. Preclinical evaluation of three polyspecific antivenoms against the venom of Echis ocellatus: Neutralization of toxic activities and antivenomics.

    PubMed

    Calvete, Juan J; Arias, Ana Silvia; Rodríguez, Yania; Quesada-Bernat, Sarai; Sánchez, Laura V; Chippaux, Jean Philippe; Pla, Davinia; Gutiérrez, José María

    2016-09-01

    Snakebite envenoming has a heavy burden in the public health in sub-Saharan Africa. The viperid species Echis ocellatus (carpet viper or saw-scaled viper) is the medically most important snake in the savannahs of western sub-Saharan Africa. Several antivenoms are being distributed and used in this region for the treatment of envenomings by E. ocellatus, but the preclinical efficacy of some of these antivenoms has not been assessed. The present study evaluated the preclinical efficacy against E. ocellatus venom of three polyspecific antivenoms: (a) Snake Venom Antiserum (Pan Africa), manufactured by Premium Serums and Vaccines (India); (b) Snake Venom Antiserum (Africa), manufactured by VINS Bioproducts (India); and (c) Antivipmyn(®) Africa, manufactured by Instituto Bioclon (Mexico). Antivenomics analysis revealed the ability of the three antivenoms to immunocapture the majority of components of the venoms of E. ocellatus from Cameroon, Nigeria and Mali, although their maximal immunocapturing capability varied. Bioclon and Premium Serums antivenoms were effective in the neutralization of lethal, hemorrhagic and in vitro coagulant activities of the venom of E. ocellatus from Cameroon, albeit with different potencies. VINS antivenom neutralized hemorrhagic activity of this venom, but failed to neutralize lethality at the highest antivenom dose tested, and had a low neutralizing efficacy against in vitro coagulant effect. PMID:27377229

  9. Tissue Plasminogen Activator Neurotoxicity is Neutralized by Recombinant ADAMTS 13

    PubMed Central

    Fan, Mengchen; Xu, Haochen; Wang, Lixiang; Luo, Haiyu; Zhu, Ximin; Cai, Ping; Wei, Lixiang; Lu, Lu; Cao, Yongliang; Ye, Rong; Fan, Wenying; Zhao, Bing-Qiao

    2016-01-01

    Tissue plasminogen activator (tPA) is an effective treatment for ischemic stroke, but its neurotoxicity is a significant problem. Here we tested the hypothesis that recombinant ADAMTS 13 (rADAMTS 13) would reduce tPA neurotoxicity in a mouse model of stroke. We show that treatment with rADAMTS 13 in combination with tPA significantly reduced infarct volume compared with mice treated with tPA alone 48 hours after stroke. The combination treatment significantly improved neurological deficits compared with mice treated with tPA or vehicle alone. These neuroprotective effects were associated with significant reductions in fibrin deposits in ischemic vessels and less severe cell death in ischemic brain. The effect of rADAMTS13 on tPA neurotoxicity was mimicked by the N-methyl-D-aspartate (NMDA) receptor antagonist M-801, and was abolished by injection of NMDA. Moreover, rADAMTS 13 prevents the neurotoxicity effect of tPA, by blocking its interaction with the NMDA receptor NR2B and the attendant phosphorylation of NR2B and activation of ERK1/2. Finally, the NR2B-specific NMDA receptor antagonist ifenprodil abolished tPA neurotoxicity and rADAMTS 13 treatment had no further beneficial effect. Our data suggest that the combination of rADAMTS 13 and tPA may provide a novel treatment of ischemic stroke by diminishing the neurotoxic effects of exogenous tPA. PMID:27181025

  10. EEG source activity during processing of neutral stimuli in subjects with anxiety disorders.

    PubMed

    Gmaj, Bartłomiej; Januszko, Piotr; Kamiński, Jan; Drozdowicz, Ewa; Kopera, Maciej; Wołyńczyk-Gmaj, Dorota; Szelenberger, Waldemar; Wojnar, Marcin

    2016-01-01

    Anxiety disorders are a social problem due to their prevalence and consequences. It is crucial to explore the influence of anxiety on cognitive processes. In this study we recorded EEG activity from 73 subjects (35 patients, 38 controls, matched for age and education) during performance of the Continuous Attention Task. We used low resolution electromagnetic tomography (LORETA) for evaluation of mechanisms of impaired cognitive performance in anxiety disorders. Analysis showed that patients with anxiety disorders committed more errors than the controls, had a short latency of P300 and higher amplitude of ERPs at all steps of stimulus processing. Furthermore, we showed that there was a relationship between the scores of Hamilton Anxiety Scale and Beck Depression Inventory, and amplitudes and latencies of ERPs. The results of LORETA analysis showed that enhanced neural responses were found within circuits mediating visual information processing, sustained attention and anxiety. Also, we found higher current density within areas playing an important role in the brain fear network - anterior cingulate and anterior part of insula. Electrophysiological neuroimaging showed greater recruitment of cognitive resources in anxiety disorders, evidenced by higher current density and activation of greater number of brain areas. Despite the strategy employed to compensate for cognitive problems, the anxiety patients did not achieve the same performance as controls. Present study demonstrates that anxiety disorders influence processing of neutral stimuli and this influence is observable at both behavioral and electrophysiological level. The data suggests instability of neural systems responsible for information selection, working memory, engagement and focusing of attention.

  11. EEG source activity during processing of neutral stimuli in subjects with anxiety disorders.

    PubMed

    Gmaj, Bartłomiej; Januszko, Piotr; Kamiński, Jan; Drozdowicz, Ewa; Kopera, Maciej; Wołyńczyk-Gmaj, Dorota; Szelenberger, Waldemar; Wojnar, Marcin

    2016-01-01

    Anxiety disorders are a social problem due to their prevalence and consequences. It is crucial to explore the influence of anxiety on cognitive processes. In this study we recorded EEG activity from 73 subjects (35 patients, 38 controls, matched for age and education) during performance of the Continuous Attention Task. We used low resolution electromagnetic tomography (LORETA) for evaluation of mechanisms of impaired cognitive performance in anxiety disorders. Analysis showed that patients with anxiety disorders committed more errors than the controls, had a short latency of P300 and higher amplitude of ERPs at all steps of stimulus processing. Furthermore, we showed that there was a relationship between the scores of Hamilton Anxiety Scale and Beck Depression Inventory, and amplitudes and latencies of ERPs. The results of LORETA analysis showed that enhanced neural responses were found within circuits mediating visual information processing, sustained attention and anxiety. Also, we found higher current density within areas playing an important role in the brain fear network - anterior cingulate and anterior part of insula. Electrophysiological neuroimaging showed greater recruitment of cognitive resources in anxiety disorders, evidenced by higher current density and activation of greater number of brain areas. Despite the strategy employed to compensate for cognitive problems, the anxiety patients did not achieve the same performance as controls. Present study demonstrates that anxiety disorders influence processing of neutral stimuli and this influence is observable at both behavioral and electrophysiological level. The data suggests instability of neural systems responsible for information selection, working memory, engagement and focusing of attention. PMID:27102920

  12. Bifurcation Analysis for Neural Networks in Neutral Form

    NASA Astrophysics Data System (ADS)

    Chen, Hong-Bing; Sun, Xiao-Ke

    2016-06-01

    In this paper, a system of neural networks in neutral form with time delay is investigated. Further, by introducing delay τ as a bifurcation parameter, it is found that Hopf bifurcation occurs when τ is across some critical values. The direction of the Hopf bifurcations and the stability are determined by using normal form method and center manifold theory. Next, the global existence of periodic solution is established by using a global Hopf bifurcation result. Finally, an example is given to support the theoretical predictions.

  13. Emittance Analysis of the DIII-D Neutral Beam Source

    NASA Astrophysics Data System (ADS)

    Lopez, N. A.; Crowley, B.

    2014-10-01

    In a high powered neutral beam system ions are extracted from a low temperature plasma, through apertures in the arc chamber, by application of a potential to an external electrode. It has been determined that to increase the beam energy of the DIII-D neutral beam system beyond 95 keV the accelerator must be reconfigured to avoid excessive electrical breakdown in the grid gaps. Deciding exactly what modifications are to be made requires modeling and experimental effort. A basic problem is to find a geometry with which the extracted beam is intense, low divergence, free of aberrations, and does not strike the focusing electrodes. We present the results of modeling proposed reconfigurations to the accelerator geometry and source conditions. The quality of the beam produced from the various accelerator configurations is quantified through metrics such as the beam emittance and the average divergence per beamlet. By comparing the beam quality and power delivered for each proposed reconfiguration an optimal design is selected and recommended. Work supported in part by the National Undergraduate Fellowship Program in Plasma Physics and Fusion Energy Sciences and the US DOE under DE-FG02-94ER54235, DE-FC02-04ER54698.

  14. Neutral-Line Magnetic Shear and Enhanced Coronal Heating in Solar Active Regions

    NASA Technical Reports Server (NTRS)

    Falconer, D. A.; Moore, R. L.; Porter, J. G.; Gary, G. A.; Shimizu, T.

    1997-01-01

    By examining the magnetic structure at sites in the bright coronal interiors of active regions that are not flaring but exhibit persistent strong coronal heating, we establish some new characteristics of the magnetic origins of this heating. We have examined the magnetic structure of these sites in five active regions, each of which was well observed by both the Yohkoh SXT and the Marshall Space Flight Center Vector Magnetograph and showed strong shear in its magnetic field along part of at least one neutral line (polarity inversion). Thus, we can assess whether this form of nonpotential field structure in active regions is a characteristic of the enhanced coronal heating and vice versa. From 27 orbits' worth of Yohkoh SXT images of the five active regions, we have obtained a sample of 94 persistently bright coronal features (bright in all images from a given orbit), 40 long (greater than or approximately equals 20,000 km) neutral-line segments having strong magnetic shear throughout (shear angle greater than 45 deg), and 39 long neutral-line segments having weak magnetic shear throughout (shear angle less than 45 deg). From this sample, we find that: (1) all of our persistently bright coronal features are rooted in magnetic fields that are stronger than 150 G; (2) nearly all (95%) of these enhanced coronal features are rooted near neutral lines (closer than 10,000 km); (3) a great majority (80%) of the bright features are rooted near strong-shear portions of neutral lines; (4) a great majority (85%) of long strong-shear segments of neutral lines have persistently bright coronal features rooted near them; (5) a large minority (40%) of long weak-shear segments of neutral lines have persistently bright coronal features rooted near them; and (6) the brightness of a persistently bright Coronal feature often changes greatly over a few hours. From these results, we conclude that most persistent enhanced heating of coronal loops in active regions: (1) requires the

  15. Neutralizing activities of caprine antibodies towards conserved regions of the HCV envelope glycoprotein E2

    PubMed Central

    2011-01-01

    Anti HCV vaccine is not currently available and the present antiviral therapies fail to cure approximately half of the treated HCV patients. This study was designed to assess the immunogenic properties of genetically conserved peptides derived from the C-terminal region of HVR-1 and test their neutralizing activities in a step towards developing therapeutic and/or prophylactic immunogens against HCV infection. Antibodies were generated by vaccination of goats with synthetic peptides derived from HCV E2. Viral neutralizing capacity of the generated anti E2 antibodies was tested using in vitro assays. Goats immunized with E2 synthetic peptides termed p412 [a.a 412-419], p430 [a.a 430-447] and p517 [a.a 517-531] generated high titers of antibody responses 2 to 4.5 fold higher than comparable titers of antibodies to the same epitopes in chronic HCV patients. In post infection experiments of native HCV into cultured Huh7.5 cells anti p412 and anti p 517 were proven to be neutralizing to HCV genotype 4a from patients' sera (87.5% and 75% respectively). On the contrary anti p430 exhibited weak viral neutralization capacity on the same samples (31.25%). Furthermore Ab mixes containing anti p430 exhibited reduced viral neutralization properties. From these experiments one could predict that neutralization by Abs towards different E2-epitopes varies considerably and success in the enrichment of neutralization epitope-specific antibodies may be accompanied by favorable results in combating HCV infection. Also, E2 conserved peptides p517 and p412 represent potential components of a candidate peptide vaccine against HCV infection. PMID:21819575

  16. A plaque assay for malignant catarrhal fever virus and virus neutralizing activity.

    PubMed

    Hazlett, D T

    1980-05-01

    A cell-free strain of malignant catarrhal fever virus which produced a readily recognizable cytopathic effect was obtained by serial passage of the virus in a rabbit kidney cell line. Plaque assay of the virus was more rapid and gave higher titres 11 days postinoculation than tube titration, but the latter advantage decreased with a longer incubation period. Plaques were clear with sharp edges and measured 0.5 to 2 mm in diameter after 15 days. A plaque neutralization test was developed and successfully employed for the titration of malignant catarrhal fever virus neutralizing activity in the sera and nasal secretions of blue wildebeest.

  17. Neutralization of feline infectious peritonitis virus: preparation of monoclonal antibody that shows cell tropism in neutralizing activity after viral absorption into the cells.

    PubMed

    Kida, K; Hohdatsu, T; Kashimoto-Tokunaga, J; Koyama, H

    2000-01-01

    Feline infectious peritonitis virus (FIPV) infection of feline macro-phages is enhanced by mouse anti-FIPV monoclonal antibody (MAb). This anti-body-dependent enhancement (ADE) of FIPV infection is dependent on mouse MAb subclass, and MAb of IgG2a subclass has a strong ADE activity. Furthermore, MAb showing strong neutralizing activity in Felis catus whole fetus (fcwf-4) cells and Crandell feline kidney (CrFK) cells shows strong enhancing activity in feline macrophages, indicating that the neutralizing epitope and the enhancing epitope are closely related. In this study, we prepared MAb FK50-4 that showed a strong neutralizing activity in feline macrophages, despite the fact that the MAb belonged to the IgG2a subclass. However, MAb FK50-4 did not exhibit neutralizing activity in CrFK cells or fcwf-4 cells, thus showing a very unusual property. MAb FK50-4 recognized FIPV small integral membrane glycoprotein (M protein). Even when feline macrophages were pretreated with MAb FK50-4 prior to FIPV inoculation, this antibody prevented FIPV infection. This reaction disappeared after treatment of FK50-4 with protein A. The neutralizing activity of FK50-4 was also effective on feline macrophages after the cells were inoculated with FIPV. These findings indicated that the FIPV replication mechanism differs between feline macrophages and CrFK/fcwf-4 cells and that a neutralizing epitope that can prevent FIPV infection of feline macrophages after viral absorption is present on M protein.

  18. Chemical acceleration of a neutral granulated blast-furnace slag activated by sodium carbonate

    SciTech Connect

    Kovtun, Maxim Kearsley, Elsabe P. Shekhovtsova, Julia

    2015-06-15

    This paper presents results of a study on chemical acceleration of a neutral granulated blast-furnace slag activated using sodium carbonate. As strength development of alkali-activated slag cements containing neutral GBFS and sodium carbonate as activator at room temperature is known to be slow, three accelerators were investigated: sodium hydroxide, ordinary Portland cement and a combination of silica fume and slaked lime. In all cements, the main hydration product is C–(A)–S–H, but its structure varies between tobermorite and riversideite depending on the accelerator used. Calcite and gaylussite are present in all systems and they were formed due to either cation exchange reaction between the slag and the activator, or carbonation. With accelerators, compressive strength up to 15 MPa can be achieved within 24 h in comparison to 2.5 MPa after 48 h for a mix without an accelerator.

  19. Charge separation promoted activation of molecular oxygen by neutral gold clusters.

    PubMed

    Woodham, Alex P; Meijer, Gerard; Fielicke, André

    2013-02-01

    Gold nanoparticles and sub-nanoparticles famously act as highly efficient and selective low-temperature oxidation catalysts with molecular oxygen, in stark contrast to the nobility of the bulk phase. The origins of this activity and the nature of the active species remain open questions. Gas-phase studies of isolated gold clusters hold promise for disentangling these problems. Here we address the interaction of neutral gold clusters (Au(n); 4 ≤ n ≤ 21) with molecular oxygen by probing the highly characteristic O-O vibrational stretch frequencies. This reveals that for selected cluster sizes the oxygen is highly activated with respect to the free moiety. Complementary quantum chemical calculations provide evidence for substantial electron transfer to the O(2) unit and concomitant rearrangement of the parent gold cluster structure upon binding and activation. This gives evidence for a model of the interaction between neutral gold clusters and molecular oxygen.

  20. Prefusion F-specific antibodies determine the magnitude of RSV neutralizing activity in human sera.

    PubMed

    Ngwuta, Joan O; Chen, Man; Modjarrad, Kayvon; Joyce, M Gordon; Kanekiyo, Masaru; Kumar, Azad; Yassine, Hadi M; Moin, Syed M; Killikelly, April M; Chuang, Gwo-Yu; Druz, Aliaksandr; Georgiev, Ivelin S; Rundlet, Emily J; Sastry, Mallika; Stewart-Jones, Guillaume B E; Yang, Yongping; Zhang, Baoshan; Nason, Martha C; Capella, Cristina; Peeples, Mark E; Ledgerwood, Julie E; McLellan, Jason S; Kwong, Peter D; Graham, Barney S

    2015-10-14

    Respiratory syncytial virus (RSV) is estimated to claim more lives among infants <1 year old than any other single pathogen, except malaria, and poses a substantial global health burden. Viral entry is mediated by a type I fusion glycoprotein (F) that transitions from a metastable prefusion (pre-F) to a stable postfusion (post-F) trimer. A highly neutralization-sensitive epitope, antigenic site Ø, is found only on pre-F. We determined what fraction of neutralizing (NT) activity in human sera is dependent on antibodies specific for antigenic site Ø or other antigenic sites on F in healthy subjects from ages 7 to 93 years. Adsorption of individual sera with stabilized pre-F protein removed >90% of NT activity and depleted binding antibodies to both F conformations. In contrast, adsorption with post-F removed ~30% of NT activity, and binding antibodies to pre-F were retained. These findings were consistent across all age groups. Protein competition neutralization assays with pre-F mutants in which sites Ø or II were altered to knock out binding of antibodies to the corresponding sites showed that these sites accounted for ~35 and <10% of NT activity, respectively. Binding competition assays with monoclonal antibodies (mAbs) indicated that the amount of site Ø-specific antibodies correlated with NT activity, whereas the magnitude of binding competed by site II mAbs did not correlate with neutralization. Our results indicate that RSV NT activity in human sera is primarily derived from pre-F-specific antibodies, and therefore, inducing or boosting NT activity by vaccination will be facilitated by using pre-F antigens that preserve site Ø.

  1. Prefusion F–specific antibodies determine the magnitude of RSV neutralizing activity in human sera

    PubMed Central

    Ngwuta, Joan O.; Chen, Man; Modjarrad, Kayvon; Joyce, M. Gordon; Kanekiyo, Masaru; Kumar, Azad; Yassine, Hadi M.; Moin, Syed M.; Killikelly, April M.; Chuang, Gwo-Yu; Druz, Aliaksandr; Georgiev, Ivelin S.; Rundlet, Emily J.; Sastry, Mallika; Stewart-Jones, Guillaume B. E.; Yang, Yongping; Zhang, Baoshan; Nason, Martha C.; Capella, Cristina; Peeples, Mark E.; Ledgerwood, Julie E.; McLellan, Jason S.; Kwong, Peter D.; Graham, Barney S.

    2015-01-01

    Respiratory syncytial virus (RSV) is estimated to claim more lives among infants <1 year old than any other single pathogen, except malaria, and poses a substantial global health burden. Viral entry is mediated by a type I fusion glycoprotein (F) that transitions from a metastable prefusion (pre-F) to a stable postfusion (post-F) trimer. A highly neutralization-sensitive epitope, antigenic site Ø, is found only on pre-F. We determined what fraction of neutralizing (NT) activity in human sera is dependent on antibodies specific for antigenic site Ø or other antigenic sites on F in healthy subjects from ages 7 to 93 years. Adsorption of individual sera with stabilized pre-F protein removed >90% of NT activity and depleted binding antibodies to both F conformations. In contrast, adsorption with post-F removed ~30% of NT activity, and binding antibodies to pre-F were retained. These findings were consistent across all age groups. Protein competition neutralization assays with pre-F mutants in which sites Ø or II were altered to knock out binding of antibodies to the corresponding sites showed that these sites accounted for ~35 and <10% of NT activity, respectively. Binding competition assays with monoclonal antibodies (mAbs) indicated that the amount of site Ø–specific antibodies correlated with NT activity, whereas the magnitude of binding competed by site II mAbs did not correlate with neutralization. Our results indicate that RSV NT activity in human sera is primarily derived from pre-F–specific antibodies, and therefore, inducing or boosting NT activity by vaccination will be facilitated by using pre-F antigens that preserve site Ø. PMID:26468324

  2. Activity Coefficient Derivatives of Ternary Systems Based on Scatchard's Neutral Electrolyte description

    SciTech Connect

    Miller, D G

    2007-05-16

    Activity coefficient derivatives with respect to molality are presented for the Scatchard Neutral Electrolyte description of a ternary common-ion electrolyte system. These quantities are needed for the calculation of 'diffusion Onsager coefficients' and in turn for tests of the Onsager Reciprocal Relations in diffusion. The usually-omitted b{sub 23} term is included. The direct SNE binary approximations and a further approximation are discussed. Binary evaluation strategies other than constant ionic strength are considered.

  3. Accumulated Bending Energy Elicits Neutral Sphingomyelinase Activity in Human Red Blood Cells

    PubMed Central

    López, David J.; Egido-Gabas, Meritxell; López-Montero, Iván; Busto, Jon V.; Casas, Josefina; Garnier, Marie; Monroy, Francisco; Larijani, Banafshé; Goñi, Félix M.; Alonso, Alicia

    2012-01-01

    We propose that accumulated membrane bending energy elicits a neutral sphingomyelinase (SMase) activity in human erythrocytes. Membrane bending was achieved by osmotic or chemical processes, and SMase activity was assessed by quantitative thin-layer chromatography, high-performance liquid chromatography, and electrospray ionization-mass spectrometry. The activity induced by hypotonic stress in erythrocyte membranes had the pH dependence, ion dependence, and inhibitor sensitivity of mammalian neutral SMases. The activity caused a decrease in SM contents, with a minimum at 6 min after onset of the hypotonic conditions, and then the SM contents were recovered. We also elicited SMase activity by adding lysophosphatidylcholine externally or by generating it with phospholipase A2. The same effect was observed upon addition of chlorpromazine or sodium deoxycholate at concentrations below the critical micellar concentration, and even under hypertonic conditions. A unifying factor of the various agents that elicit this SMase activity is the accumulated membrane bending energy. Both hypo-and hypertonic conditions impose an increased curvature, whereas the addition of surfactants or phospholipase A2 activation increases the outer monolayer area, thus leading to an increased bending energy. The fact that this latent SMase activity is tightly coupled to the membrane bending properties suggests that it may be related to the general phenomenon of stress-induced ceramide synthesis and apoptosis. PMID:22824271

  4. Alkaline, acid, and neutral phosphatase activities are induced during development in Myxococcus xanthus.

    PubMed Central

    Weinberg, R A; Zusman, D R

    1990-01-01

    One of the signals that has been reported to be important in stimulating fruiting body formation of Myxococcus xanthus is starvation for phosphate. We therefore chose to study phosphatase activity during M. xanthus development. Many phosphatases can cleave the substrate p-nitrophenol phosphate. Using this substrate in buffers at various pHs, we obtained a profile of phosphatase activities during development and germination of M. xanthus. These experiments indicated that there are five patterns of phosphatase activity in M. xanthus: two vegetative and three developmental. The two uniquely vegetative activities have pH optima at 7.2 and 8.5. Both require magnesium and both are inhibited by the reducing agent dithiothreitol. The developmental (spores) patterns of activity have pH optima of 5.2, 7.2, and 8.5. All three activities are Mg independent. Only the alkaline phosphatase activity is inhibited by dithiothreitol. The acid phosphatase activity is induced very early in development, within the first 2 to 4 h. Both the neutral and alkaline phosphatase Mg-independent activities are induced much later, about the time that myxospores become evident (24 to 30 h). The three activities are greatly diminished upon germination; however, the kinetics of loss differ for all three. The acid phosphatase activity declines very rapidly, the neutral activity begins to decline only after spores begin to convert to rods, and the alkaline phosphatase activity remains high until the time the cells begin to divide. All three developmental activities were measured in the developmental signalling mutants carrying asg, csg, and dsg. The pattern of expression obtained in the mutants was consistent with that of other developmentally regulated genes which exhibit similar patterns of expression during development. The ease with which phosphatases can be assayed should make the activities described in this report useful biochemical markers of stages of both fruiting body formation and

  5. Classification of illicit heroin by UPLC-Q-TOF analysis of acidic and neutral manufacturing impurities.

    PubMed

    Liu, Cuimei; Hua, Zhendong; Bai, Yanping

    2015-12-01

    The illicit manufacture of heroin results in the formation of trace levels of acidic and neutral manufacturing impurities that provide valuable information about the manufacturing process used. In this work, a new ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF) method; that features high resolution, mass accuracy and sensitivity for profiling neutral and acidic heroin manufacturing impurities was developed. After the UPLC-Q-TOF analysis, the retention times and m/z data pairs of acidic and neutral manufacturing impurities were detected, and 19 peaks were found to be evidently different between heroin samples from "Golden Triangle" and "Golden Crescent". Based on the data set of these 19 impurities in 150 authentic heroin samples, classification of heroin geographic origins was successfully achieved utilizing partial least squares discriminant analysis (PLS-DA). By analyzing another data set of 267 authentic heroin samples, the developed discrimiant model was validated and proved to be accurate and reliable.

  6. Thermospheric topside neutral density, ionospheric anomalous electric field and resistivity measurements by active experiment at EISCAT

    NASA Astrophysics Data System (ADS)

    Kosch, Michael; Ogawa, Yasunobu; Rietveld, Michael; Blagoveshchenskaya, Nataly; Yamazaki, Yosuke

    2016-07-01

    We have developed an active ground-based technique to estimate the topside thermospheric neutral density as well as topside ionospheric anomalous electric field and resistivity at EISCAT, combining the EISCAT UHF radar, HF heater and optics. When pumping the ionosphere the F-region electron temperature is significantly raised, increasing the upward plasma pressure gradient in the topside ionosphere, resulting in observed ion up-flow along the magnetic field line. Simultaneously, pump-induced suprathermal electrons produce artificial optical emissions. Using the modified ion-momentum equation, the thermospheric neutral density is estimated. Alternatively, using the MSIS model the field-aligned anomalous electric field is estimated. From the optical data the suprathermal electron flux is estimated, giving an estimate of the anomalous resistivity. Results from recent observations at EISCAT are presented.

  7. Highly active cobalt phosphate and borate based oxygen evolving catalysts operating in neutral and natural waters

    SciTech Connect

    Esswein, AJ; Surendranath, Y; Reece, SY; Nocera, DG

    2011-02-01

    A high surface area electrode is functionalized with cobalt-based oxygen evolving catalysts (Co-OEC = electrodeposited from pH 7 phosphate, Pi, pH 8.5 methylphosphonate, MePi, and pH 9.2 borate electrolyte, Bi). Co-OEC prepared from MePi and operated in Pi and Bi achieves a current density of 100 mA cm(-2) for water oxidation at 442 and 363 mV overpotential, respectively. The catalyst retains activity in near-neutral pH buffered electrolyte in natural waters such as those from the Charles River (Cambridge, MA) and seawater (Woods Hole, MA). The efficacy and ease of operation of anodes functionalized with Co-OEC at appreciable current density together with its ability to operate in near neutral pH buffered natural water sources bodes well for the translation of this catalyst to a viable renewable energy storage technology.

  8. Kinetics of positive ions and electrically neutral active particles in afterglow in neon at low pressure

    SciTech Connect

    Pejović, Milić M. Nešić, Nikola T.; Pejović, Momčilo M.

    2014-04-15

    Kinetics of positive ions and electrically neutral active particles formed during breakdown and successive discharge in neon-filled tube at 6.6 millibars pressure had been analyzed. This analysis was performed on the basis of mean value of electrical breakdown time delay t{sup ¯}{sub d} dependence on afterglow period τ (memory curve). It was shown that positive ions are present in the 1μs < τ < 30 ms interval, which is manifested through t{sup ¯}{sub d} slow increase with the increase of τ. A rapid t{sup ¯}{sub d} increase in the 30 ms < τ < 3 s interval is a consequence of significant decrease of positive ions concentration and dominant role in breakdown initiation have ground state nitrogen atoms, which further release secondary electrons from the cathode by catalytic recombination process. These atoms are formed during discharge by dissociation of ground state nitrogen molecules that are present as impurities in neon. For τ > 3 s, breakdown is initiated by cosmic rays and natural radioactivity. The increase of discharge current leads to decrease of t{sup ¯}{sub d} due to the increase of positive ions concentration in inter electrode gap. The increase of applied voltage also decreases t{sup ¯}{sub d} for τ > 30 ms due to the increase of the probability for initial electron to initiate breakdown. The presence of UV radiation leads to the decrease of t{sup ¯}{sub d} due to the increased electron yield caused by photoelectrons. The influence of photoelectrons on breakdown initiation can be noticed for τ > 0.1 ms, while they dominantly determine t{sup ¯}{sub d} for τ > 30 ms.

  9. Kinetics of positive ions and electrically neutral active particles in afterglow in neon at low pressure

    NASA Astrophysics Data System (ADS)

    Pejović, Milić M.; Nešić, Nikola T.; Pejović, Momčilo M.

    2014-04-01

    Kinetics of positive ions and electrically neutral active particles formed during breakdown and successive discharge in neon-filled tube at 6.6 millibars pressure had been analyzed. This analysis was performed on the basis of mean value of electrical breakdown time delay t¯d dependence on afterglow period τ (memory curve). It was shown that positive ions are present in the 1μs < τ < 30 ms interval, which is manifested through t ¯d slow increase with the increase of τ. A rapid t¯d increase in the 30 ms < τ < 3 s interval is a consequence of significant decrease of positive ions concentration and dominant role in breakdown initiation have ground state nitrogen atoms, which further release secondary electrons from the cathode by catalytic recombination process. These atoms are formed during discharge by dissociation of ground state nitrogen molecules that are present as impurities in neon. For τ > 3 s, breakdown is initiated by cosmic rays and natural radioactivity. The increase of discharge current leads to decrease of t¯d due to the increase of positive ions concentration in inter electrode gap. The increase of applied voltage also decreases t¯d for τ > 30 ms due to the increase of the probability for initial electron to initiate breakdown. The presence of UV radiation leads to the decrease of t¯d due to the increased electron yield caused by photoelectrons. The influence of photoelectrons on breakdown initiation can be noticed for τ > 0.1 ms, while they dominantly determine t¯d for τ > 30 ms.

  10. Activity and stability trends of perovskite oxides for oxygen evolution catalysis at neutral pH.

    PubMed

    Han, Binghong; Risch, Marcel; Lee, Yueh-Lin; Ling, Chen; Jia, Hongfei; Shao-Horn, Yang

    2015-09-21

    Perovskite oxides (ABO3) have been studied extensively to promote the kinetics of the oxygen evolution reaction (OER) in alkaline electrolytes. However, developing highly active catalysts for OER at near-neutral pH is desirable for many photoelectrochemical/electrochemical devices. In this paper, we systematically studied the activity and stability of well-known perovskite oxides for OER at pH 7. Previous activity descriptors established for perovskite oxides at pH 13, such as having an eg occupancy close to unity or having an O p-band center close to Fermi level, were shown to scale with OER activity at pH 7. Stability was a greater challenge at pH 7 than at pH 13, where two different modes of instability were identified from combined transmission electron microscopy and density functional theory analyses. Perovskites with O p-band close to Fermi level showed leaching of A-site atoms and surface amorphization under all overpotentials examined at pH 7, while those with O p-band far from Fermi level were stable under low OER current/potential but became unstable at high current/potential accompanied by leaching of B-site atoms. Therefore, efforts are needed to enhance the activity and stability of perovskites against A-site or B-site loss if used at neutral pH. PMID:26271910

  11. Activity and stability trends of perovskite oxides for oxygen evolution catalysis at neutral pH.

    PubMed

    Han, Binghong; Risch, Marcel; Lee, Yueh-Lin; Ling, Chen; Jia, Hongfei; Shao-Horn, Yang

    2015-09-21

    Perovskite oxides (ABO3) have been studied extensively to promote the kinetics of the oxygen evolution reaction (OER) in alkaline electrolytes. However, developing highly active catalysts for OER at near-neutral pH is desirable for many photoelectrochemical/electrochemical devices. In this paper, we systematically studied the activity and stability of well-known perovskite oxides for OER at pH 7. Previous activity descriptors established for perovskite oxides at pH 13, such as having an eg occupancy close to unity or having an O p-band center close to Fermi level, were shown to scale with OER activity at pH 7. Stability was a greater challenge at pH 7 than at pH 13, where two different modes of instability were identified from combined transmission electron microscopy and density functional theory analyses. Perovskites with O p-band close to Fermi level showed leaching of A-site atoms and surface amorphization under all overpotentials examined at pH 7, while those with O p-band far from Fermi level were stable under low OER current/potential but became unstable at high current/potential accompanied by leaching of B-site atoms. Therefore, efforts are needed to enhance the activity and stability of perovskites against A-site or B-site loss if used at neutral pH.

  12. Novel pyrazole derivatives as neutral CB₁ antagonists with significant activity towards food intake.

    PubMed

    Manca, Ilaria; Mastinu, Andrea; Olimpieri, Francesca; Falzoi, Matteo; Sani, Monica; Ruiu, Stefania; Loriga, Giovanni; Volonterio, Alessandro; Tambaro, Simone; Bottazzi, Mirko Emilio Heiner; Zanda, Matteo; Pinna, Gérard Aimè; Lazzari, Paolo

    2013-04-01

    In spite of rimonabant's withdrawal from the European market due to its adverse effects, interest in the development of drugs based on CB1 antagonists is revamping on the basis of the peculiar properties of this class of compounds. In particular, new strategies have been proposed for the treatment of obesity and/or related risk factors through CB1 antagonists, i.e. by the development of selectively peripherally acting agents or by the identification of neutral CB1 antagonists. New compounds based on the lead CB1 antagonist/inverse agonist rimonabant have been synthesized with focus on obtaining neutral CB1 antagonists. Amongst the new derivatives described in this paper, the mixture of the two enantiomers (±)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-3-(2-cyclohexyl-1-hydroxyethyl)-4-methyl-1H-pyrazole ((±)-5), and compound 5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-3-[(Z)-2-cyclohexyl-1-fluorovinyl]-4-methyl-1H-pyrazole ((Z)-6), showed interesting pharmacological profiles. According to the preliminary pharmacological evaluation, these novel pyrazole derivatives showed in fact both neutral CB1 antagonism behaviour and significant in vivo activity towards food intake.

  13. Neutralization activity of patient sera collected during the 2008-2009 Chikungunya outbreak in Thailand.

    PubMed

    Kishishita, Natsuko; Sasayama, Mikiko; Takeda, Naokazu; Sa-Ngasang, Areerat; Anuegoonpipat, Atchareeya; Anantapreecha, Surapee

    2015-01-01

    Chikungunya virus (CHIKV) infection typically causes fever, rash, myalgia, and arthralgia and sometimes results in recurrent joint pain or, in severe cases, neurological disorders or death. How CHIKV infection leads to prolonged or severe symptoms is still not well understood. In this study, we examined the neutralization (NT) titer of 98 serum samples collected from patients during the 2008-2009 chikungunya outbreak in Thailand. While all serum samples showed neutralizing activity, virus was detected in 58% of the serum samples. When we analyzed a possible association between virus and antibody titers and the presence of typical symptoms of CHIKV infection, fever and joint pain, there was no significant association except that the number of patients with fever was over three times more than the number of those without fever when CHIKV was detectable in serum. This study indicates that although neutralizing antibody is critical to eliminate CHIKV, it appears not to be the main factor associated with clinical symptoms in some cases, so that other aspects of immune responses, such as those involving proinflammatory mediators and adaptive immune cells, should be considered altogether.

  14. Toward Effective HIV Vaccination INDUCTION OF BINARY EPITOPE REACTIVE ANTIBODIES WITH BROAD HIV NEUTRALIZING ACTIVITY

    SciTech Connect

    Nishiyama, Yasuhiro; Planque, Stephanie; Mitsuda, Yukie; Nitti, Giovanni; Taguchi, Hiroaki; Jin, Lei; Symersky, Jindrich; Boivin, Stephane; Sienczyk, Marcin; Salas, Maria; Hanson, Carl V.; Paul, Sudhir

    2009-11-23

    We describe murine monoclonal antibodies (mAbs) raised by immunization with an electrophilic gp120 analog (E-gp120) expressing the rare ability to neutralize genetically heterologous human immunodeficiency virus (HIV) strains. Unlike gp120, E-gp120 formed covalent oligomers. The reactivity of gp120 and E-gp120 with mAbs to reference neutralizing epitopes was markedly different, indicating their divergent structures. Epitope mapping with synthetic peptides and electrophilic peptide analogs indicated binary recognition of two distinct gp120 regions by anti-E-gp120 mAbs, the 421-433 and 288-306 peptide regions. Univalent Fab and single chain Fv fragments expressed the ability to recognize both peptides. X-ray crystallography of an anti-E-gp120 Fab fragment revealed two neighboring cavities, the typical antigen-binding cavity formed by the complementarity determining regions (CDRs) and another cavity dominated by antibody heavy chain variable (VH) domain framework (FR) residues. Substitution of the FR cavity VH Lys-19 residue by an Ala residue resulted in attenuated binding of the 421-433 region peptide probe. The CDRs and VH FR replacement/silent mutation ratios exceeded the ratio for a random mutation process, suggesting adaptive development of both putative binding sites. All mAbs studied were derived from VH1 family genes, suggesting biased recruitment of the V gene germ line repertoire by E-gp120. The conserved 421-433 region of gp120 is essential for HIV binding to host CD4 receptors. This region is recognized weakly by the FR of antibodies produced without exposure to HIV, but it usually fails to induce adaptive synthesis of neutralizing antibodies. We present models accounting for improved CD4-binding site recognition and broad HIV neutralizing activity of the mAbs, long sought goals in HIV vaccine development.

  15. Interstellar neutral flow characteristics, composition, and interaction with the heliosphere - neutral gas and pickup ion analysis from ongoing observations and perspectives for IMAP

    NASA Astrophysics Data System (ADS)

    Moebius, E.; Bzowski, M.; Drews, C.; Frisch, P. C.; Fuselier, S. A.; Galli, A.; Gloeckler, G.; Kubiak, M. A.; Kucharek, H.; Lee, M. A.; Leonard, T.; McComas, D. J.; Park, J.; Schwadron, N.; Swaczyna, P.; Sokol, J. M.; Wood, B. E.; Wurz, P.

    2015-12-01

    The Sun's motion relative to the surrounding interstellar medium leads to an interstellar neutral (ISN) wind through the heliosphere that is moderately depleted by ionization. This situation allows remote sensing of the ISN through resonant scattering of solar UV and in-situ sampling, first via pickup ions (PUI) and most recently with direct neutral atom imaging. PUI observations have revealed the gravitational focusing cone of interstellar He and Ne as well as the composition of high ionization potential elements. After the first direct ISN He observations with Ulysses GAS, the Interstellar Boundary Explorer (IBEX) observes with high collecting power the ISN flow trajectories very close to their perihelion in Earth's orbit for H, He, O, and Ne from December through March. Meanwhile, IBEX has recorded seven years of ISN observations, with changing solar activity and varying viewing strategies. These recurring and remarkably repeatable observations allow us to consolidate the derived physical parameters and some key compositional aspects of the interstellar medium. IBEX observations provide a very precise relation between ISN flow longitude and speed via the hyperbolic trajectory equation, but with larger uncertainties separately for longitude and speed. Recent concerted studies have led to a velocity vector that is consistent between IBEX and Ulysses, with a substantially higher temperature than found previously. The fact that the IBEX He and O ISN observations contain a substantial secondary neutral contribution adds complexity to the quantitative analysis of the physical interstellar medium parameters. However, their discovery also provides invaluable insight into the interstellar plasma interaction in the outer heliosheath, which is shaped strongly by the interstellar magnetic field. The longitude range of the IBEX observations limits the precision of the ISN velocity vector. The IBEX collection power and its sensitivity to the Earth's magnetosphere limit

  16. Modeling activities on the negative-ion-based Neutral Beam Injectors of the Large Helical Device

    SciTech Connect

    Agostinetti, P.; Antoni, V.; Chitarin, G.; Pilan, N.; Serianni, G.; Veltri, P.; Cavenago, M.; Nakano, H.; Takeiri, Y.; Tsumori, K.

    2011-09-26

    At the National Institute for Fusion Science (NIFS) large-scaled negative ion sources have been widely used for the Neutral Beam Injectors (NBIs) mounted on the Large Helical Device (LHD), which is the world-largest superconducting helical system. These injectors have achieved outstanding performances in terms of beam energy, negative-ion current and optics, and represent a reference for the development of heating and current drive NBIs for ITER.In the framework of the support activities for the ITER NBIs, the PRIMA test facility, which includes a RF-drive ion source with 100 keV accelerator (SPIDER) and a complete 1 MeV Neutral Beam system (MITICA) is under construction at Consorzio RFX in Padova.An experimental validation of the codes has been undertaken in order to prove the accuracy of the simulations and the soundness of the SPIDER and MITICA design. To this purpose, the whole set of codes have been applied to the LHD NBIs in a joint activity between Consorzio RFX and NIFS, with the goal of comparing and benchmarking the codes with the experimental data. A description of these modeling activities and a discussion of the main results obtained are reported in this paper.

  17. A Correlation Between Length of Strong-Shear Neutral Lines and Total X-Ray Brightness in Active Regions

    NASA Technical Reports Server (NTRS)

    Falconer, D. A.

    1997-01-01

    From a sample of 7 MSFC vector magnetograms,of active regions and 17 Yohkoh SXT soft X-ray images of these active regions, we have found that the total x-ray brightness of an entire active region is correlated with the total length of neutral lines on which the magnetic field is both strong (less than 250 G) and strongly sheared (shear angle greater than 75 deg) in the same active region. This correlation, if not fortuitous, is additional evidence of the importance of strong-shear strong-field neutral lines to strong heating in active regions.

  18. Entamoeba histolytica: soluble and membrane-associated neutral sphingomyelinase-C and other unidentified esterase activity.

    PubMed

    Vargas-Villarreal, Javier; Palacios-Corona, Rebeca; Hernández-Luna, Carlos; Mata-Cárdenas, Benito David; Torres de la Cruz, Victor M; Cortés-Gutiérrez, Elva I; González-Salazar, Francisco; Garza-González, Jesús Norberto; Escobedo-Guajardo, Brenda Leticia; Said-Fernández, Salvador

    2010-08-01

    Sphingomyelinase (SMase) activity was measured in Entamoeba histolytica particulate and soluble subcellular fractions. The effects on SMase of incubation time, total protein concentration, pH, and several divalent cations were determined. SMase-C and other unidentified esterase activity were detected in soluble and particulate fractions. SMase-C was 94.5-96.0% higher than the unidentified esterase activity. Soluble and insoluble SMase-C specific activities increased with protein dose and incubation time. Soluble and insoluble SMase-C activities were maximum at pH 7.5 and were dependent on Mg(2+), Mn(2+), or Co(2+), and inhibited by Zn(2+), Hg(2+), Ca(2+), and EDTA. SMase-C was active in the pH range of 3-10 and its maximum activity was at pH 7.5. The soluble and insoluble SMases have remarkably similar physicochemical properties, strongly suggesting that E. histolytica has just one isoform of neutral SMase-C that had not been described before and might be essential for E. histolytica metabolism or virulence. PMID:20350542

  19. Analysis of variola and vaccinia virus neutralization assays for smallpox vaccines.

    PubMed

    Hughes, Christine M; Newman, Frances K; Davidson, Whitni B; Olson, Victoria A; Smith, Scott K; Holman, Robert C; Yan, Lihan; Frey, Sharon E; Belshe, Robert B; Karem, Kevin L; Damon, Inger K

    2012-07-01

    Possible smallpox reemergence drives research for third-generation vaccines that effectively neutralize variola virus. A comparison of neutralization assays using different substrates, variola and vaccinia (Dryvax and modified vaccinia Ankara [MVA]), showed significantly different 90% neutralization titers; Dryvax underestimated while MVA overestimated variola neutralization. Third-generation vaccines may rely upon neutralization as a correlate of protection.

  20. EFFECT OF ION-NEUTRAL COLLISIONS IN SIMULATIONS OF EMERGING ACTIVE REGIONS

    SciTech Connect

    Leake, James E.; Linton, Mark G.

    2013-02-10

    We present results of 2.5D numerical simulations of the emergence of sub-surface magnetic flux into the solar atmosphere, with emerging flux regions ranging from 10{sup 18} to 10{sup 21} Mx, representing both ephemeral and active regions. We include the presence of neutral hydrogen in the governing equations, improve upon previous models by including the ionization in the equation of state, and use a more realistic convection zone model. We find that ionization and recombination of plasma during the rise of a convection zone flux tube reduces the rise speed of the tube's axis. The presence of neutral hydrogen allows the effective flow of mass across field lines, by the addition of a Pedersen resistivity to the generalized Ohm's law, which dissipates current perpendicular to the magnetic field. This causes an increase of up to 10% in the amount of magnetic in-plane flux supplied to the corona and a reduction of up to 89% in the amount of sub-surface plasma brought up into the corona. However, it also reduces the amount of free magnetic energy supplied to the corona, and thus does not positively affect the likelihood of creating unstable coronal structures.

  1. Brain Activity in Response to Trauma-specific, Negative, and Neutral Stimuli. A fMRI Study of Recent Road Traffic Accident Survivors.

    PubMed

    Nilsen, Andre S; Blix, Ines; Leknes, Siri; Ekeberg, Øivind; Skogstad, Laila; Endestad, Tor; Østberg, Bjørn C; Heir, Trond

    2016-01-01

    Most studies of neuro-functional patterns in trauma-exposed individuals have been conducted considerable time after the traumatic event. Hence little is known about neuro-functional processing shortly after trauma-exposure. We investigated brain activity patterns in response to trauma reminders as well as neutral and negative stimuli in individuals who had recently (within 3 weeks) been involved in a road traffic accident (RTA). Twenty-three RTA survivors and 17 non-trauma-exposed healthy controls (HCs) underwent functional MRI while viewing Trauma-specific, Negative, and Neutral pictures. Data were analyzed from four a priori regions of interest, including bilateral amygdala, subcallosal cortex, and medial prefrontal cortex. In addition, we performed a whole brain analysis and functional connectivity analysis during stimulus presentation. For both groups, Negative stimuli elicited more activity in the amygdala bilaterally than did Neutral and Trauma-specific stimuli. The whole brain analysis revealed higher activation in sensory processing related areas (bilateral occipital and temporal cortices and thalamus) as well as frontal and superior parietal areas, for the RTA group compared to HC, for Trauma-specific stimuli contrasted with Neutral stimuli. We also observed higher functional connectivity for Trauma-specific stimuli, between bilateral amygdala and somatosensory areas, for the RTA group compared to controls, when contrasted with Neutral stimuli. We argue that these results might indicate an attentional sensory processing bias toward Trauma-specific stimuli for trauma exposed individuals, a result in line with findings from the post-traumatic stress disorder literature. PMID:27547195

  2. Brain Activity in Response to Trauma-specific, Negative, and Neutral Stimuli. A fMRI Study of Recent Road Traffic Accident Survivors

    PubMed Central

    Nilsen, Andre S.; Blix, Ines; Leknes, Siri; Ekeberg, Øivind; Skogstad, Laila; Endestad, Tor; Østberg, Bjørn C.; Heir, Trond

    2016-01-01

    Most studies of neuro-functional patterns in trauma-exposed individuals have been conducted considerable time after the traumatic event. Hence little is known about neuro-functional processing shortly after trauma-exposure. We investigated brain activity patterns in response to trauma reminders as well as neutral and negative stimuli in individuals who had recently (within 3 weeks) been involved in a road traffic accident (RTA). Twenty-three RTA survivors and 17 non-trauma-exposed healthy controls (HCs) underwent functional MRI while viewing Trauma-specific, Negative, and Neutral pictures. Data were analyzed from four a priori regions of interest, including bilateral amygdala, subcallosal cortex, and medial prefrontal cortex. In addition, we performed a whole brain analysis and functional connectivity analysis during stimulus presentation. For both groups, Negative stimuli elicited more activity in the amygdala bilaterally than did Neutral and Trauma-specific stimuli. The whole brain analysis revealed higher activation in sensory processing related areas (bilateral occipital and temporal cortices and thalamus) as well as frontal and superior parietal areas, for the RTA group compared to HC, for Trauma-specific stimuli contrasted with Neutral stimuli. We also observed higher functional connectivity for Trauma-specific stimuli, between bilateral amygdala and somatosensory areas, for the RTA group compared to controls, when contrasted with Neutral stimuli. We argue that these results might indicate an attentional sensory processing bias toward Trauma-specific stimuli for trauma exposed individuals, a result in line with findings from the post-traumatic stress disorder literature. PMID:27547195

  3. Measurement of the total active 8B solar neutrino flux at the Sudbury Neutrino Observatory with enhanced neutral current sensitivity.

    PubMed

    Ahmed, S N; Anthony, A E; Beier, E W; Bellerive, A; Biller, S D; Boger, J; Boulay, M G; Bowler, M G; Bowles, T J; Brice, S J; Bullard, T V; Chan, Y D; Chen, M; Chen, X; Cleveland, B T; Cox, G A; Dai, X; Dalnoki-Veress, F; Doe, P J; Dosanjh, R S; Doucas, G; Dragowsky, M R; Duba, C A; Duncan, F A; Dunford, M; Dunmore, J A; Earle, E D; Elliott, S R; Evans, H C; Ewan, G T; Farine, J; Fergani, H; Fleurot, F; Formaggio, J A; Fowler, M M; Frame, K; Fulsom, B G; Gagnon, N; Graham, K; Grant, D R; Hahn, R L; Hall, J C; Hallin, A L; Hallman, E D; Hamer, A S; Handler, W B; Hargrove, C K; Harvey, P J; Hazama, R; Heeger, K M; Heintzelman, W J; Heise, J; Helmer, R L; Hemingway, R J; Hime, A; Howe, M A; Jagam, P; Jelley, N A; Klein, J R; Kos, M S; Krumins, A V; Kutter, T; Kyba, C C M; Labranche, H; Lange, R; Law, J; Lawson, I T; Lesko, K T; Leslie, J R; Levine, I; Luoma, S; MacLellan, R; Majerus, S; Mak, H B; Maneira, J; Marino, A D; McCauley, N; McDonald, A B; McGee, S; McGregor, G; Mifflin, C; Miknaitis, K K S; Miller, G G; Moffat, B A; Nally, C W; Nickel, B G; Noble, A J; Norman, E B; Oblath, N S; Okada, C E; Ollerhead, R W; Orrell, J L; Oser, S M; Ouellet, C; Peeters, S J M; Poon, A W P; Robertson, B C; Robertson, R G H; Rollin, E; Rosendahl, S S E; Rusu, V L; Schwendener, M H; Simard, O; Simpson, J J; Sims, C J; Sinclair, D; Skensved, P; Smith, M W E; Starinsky, N; Stokstad, R G; Stonehill, L C; Tafirout, R; Takeuchi, Y; Tesić, G; Thomson, M; Thorman, M; Van Berg, R; Van de Water, R G; Virtue, C J; Wall, B L; Waller, D; Waltham, C E; Tseung, H Wan Chan; Wark, D L; West, N; Wilhelmy, J B; Wilkerson, J F; Wilson, J R; Wouters, J M; Yeh, M; Zuber, K

    2004-05-01

    The Sudbury Neutrino Observatory has precisely determined the total active (nu(x)) 8B solar neutrino flux without assumptions about the energy dependence of the nu(e) survival probability. The measurements were made with dissolved NaCl in heavy water to enhance the sensitivity and signature for neutral-current interactions. The flux is found to be 5.21 +/- 0.27(stat)+/-0.38(syst) x 10(6) cm(-2) s(-1), in agreement with previous measurements and standard solar models. A global analysis of these and other solar and reactor neutrino results yields Deltam(2)=7.1(+1.2)(-0.6) x 10(-5) eV(2) and theta=32.5(+2.4)(-2.3) degrees. Maximal mixing is rejected at the equivalent of 5.4 standard deviations.

  4. Anti-angiogenic poly-L-lysine dendrimer binds heparin and neutralizes its activity.

    PubMed

    Al-Jamal, Khuloud T; Al-Jamal, Wafa T; Kostarelos, Kostas; Turton, John A; Florence, Alexander T

    2012-01-01

    The interaction between heparin, a polyanion, and a polycationic dendrimer with a glycine core and lysine branches Gly-Lys63(NH2)64 has been investigated. Complexation was assessed by transmission electron microscopy, size and zeta potential measurements, methylene blue spectroscopy, and measuring the anti-coagulant activity of heparin in vitro and in vivo. Complete association between the heparin and the dendrimer occurred a 1:1 mass ratio (2:1 molar ratio or +/-charge ratio) with formation of quasi-spherical complexes in the size range of 99-147 nm with a negative zeta potential (-47 mV). Heparin-dendrimer (dendriplex) formation led to a concentration-dependent neutralization of the anticoagulant activity of heparin in human plasma in vitro, with complete loss of activity at a 1:1 mass ratio. The anticoagulant activity of the dendriplexes in Sprague-Dawley rats was also evaluated after subcutaneous administration with uncomplexed heparin as a comparator. The in vivo anticoagulant activity of heparin in plasma, evaluated using an antifactor Xa assay, was abolished after complexation. Measurement of [(3)H]-heparin showed that both free heparin and dendriplexes were present in plasma and in organs. Such data confirmed stably the formation of dendriplexes, which could be essential in developing novel dendrimer-based anti-angiogenic therapeutics suitable in combinatory therapeutics and theranostics. PMID:25755989

  5. Neutral sphingomyelinase activity dependent on Mg2+ and anionic phospholipids in the intraerythrocytic malaria parasite Plasmodium falciparum.

    PubMed Central

    Hanada, K; Mitamura, T; Fukasawa, M; Magistrado, P A; Horii, T; Nishijima, M

    2000-01-01

    Sphingolipid metabolism and metabolites are important in various cellular events in eukaryotes. However, little is known about their function in plasmodial parasites. Here we demonstrate that neutral sphingomyelinase (SMase) involved in the sphingomyelin (SM) catabolism is retained by the intraerythrocytic parasite Plasmodium falciparum. When assayed in a neutral pH buffer supplemented with Mg(2+) and phosphatidylserine, an activity for the release of the phosphocholine group from SM was detected in parasite-infected, but not in uninfected, erythrocyte ghosts. The SMase activity in the parasite-infected erythrocyte ghosts was enhanced markedly by anionic phospholipids including unsaturated but not saturated phosphatidylserine. Mn(2+) could not substitute for Mg(2+) to activate SMase in parasite-infected erythrocyte ghosts, whereas both Mn(2+) and Mg(2+) activated mammalian neutral SMase. The specific activity level of SMase was higher in isolated parasites than in infected erythrocyte ghosts; further fractionation of lysates of the isolated parasites showed that the activity was bound largely to the membrane fraction of the parasites. The plasmodial SMase seemed not to hydrolyse phosphatidylcholine or phosphatidylinositol. The plasmodial SMase, but not SM synthase, was sensitive to scyphostatin, an inhibitor of mammalian neutral SMase, indicating that the plasmodial activities for SM hydrolysis and SM synthesis are mediated by different catalysts. Our finding that the malaria parasites possess SMase activity might explain why the parasites seem to have an SM synthase activity but no activity to synthesize ceramide de novo. PMID:10698693

  6. Classification of illicit heroin by UPLC-Q-TOF analysis of acidic and neutral manufacturing impurities.

    PubMed

    Liu, Cuimei; Hua, Zhendong; Bai, Yanping

    2015-12-01

    The illicit manufacture of heroin results in the formation of trace levels of acidic and neutral manufacturing impurities that provide valuable information about the manufacturing process used. In this work, a new ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF) method; that features high resolution, mass accuracy and sensitivity for profiling neutral and acidic heroin manufacturing impurities was developed. After the UPLC-Q-TOF analysis, the retention times and m/z data pairs of acidic and neutral manufacturing impurities were detected, and 19 peaks were found to be evidently different between heroin samples from "Golden Triangle" and "Golden Crescent". Based on the data set of these 19 impurities in 150 authentic heroin samples, classification of heroin geographic origins was successfully achieved utilizing partial least squares discriminant analysis (PLS-DA). By analyzing another data set of 267 authentic heroin samples, the developed discrimiant model was validated and proved to be accurate and reliable. PMID:26364155

  7. Neutral buoyancy evaluation of extravehicular activity assembly of a large precision reflector

    NASA Technical Reports Server (NTRS)

    Heard, Walter L., Jr.; Lake, Mark S.

    1993-01-01

    A procedure that enables astronauts in extravehicular activity (EVA) to perform efficient on-orbit assembly of large paraboloidal precision reflectors is presented. The procedure and associated hardware are verified in simulated Og (neutral buoyancy) assembly tests of a 14m-diameter precision reflector mockup. The test article represents a precision reflector having a reflective surface which is segmented into 37 individual panels. The panels are supported on a doubly curved tetrahedral truss consisting of 315 struts. The entire truss and seven reflector panels were assembled in three hours and seven minutes by two pressure-suited test subjects. The average time to attach a panel was two minutes and three seconds. These efficient assembly times were achieved because all hardware and assembly procedures were designed to be compatible with EVA assembly capabilities.

  8. Analysis by plaque reduction neutralization assay of intertypic rotaviruses suggests that gene reassortment occurs in vivo.

    PubMed

    Hoshino, Y; Sereno, M M; Midthun, K; Flores, J; Chanock, R M; Kapikian, A Z

    1987-02-01

    The SB-1A rotavirus recovered from a diarrheic piglet in the United States is a naturally occurring intertypic rotavirus. When studied by reciprocal neutralization tests, the SB-1A virus was similar, if not identical, to the porcine Gottfried virus (serotype 4) and the porcine OSU virus (serotype 5). Analysis of reassortant viruses prepared from the SB-1A virus and the serotype 2 human DS-1 virus revealed that the antigenic specificity of the outer capsid protein VP3 of SB-1A was shared with the OSU virus, while the antigenic specificity of another outer capsid protein, VP7, of SB-1A appeared to be shared with the Gottfried virus. This suggests that SB-1A is a naturally occurring reassortant rotavirus between OSU-like and Gottfried-like porcine rotaviruses. In addition, using a genetic approach, we found evidence that the fourth gene was responsible for the predominantly one-way cross-neutralizing reactivity between canine rotavirus strain CU-1 (serotype 3) and porcine rotavirus strains SB-1A (serotypes 4 and 5) and OSU (serotype 5). Assignment of hemagglutination function to the fourth genome segment of porcine rotaviruses SB-1A and OSU and canine rotavirus CU-1 confirmed a similar previous gene assignment established for certain rotaviruses. Analysis of single gene 4 substitution reassortants confirmed our previous finding that VP3 was as potent in stimulating neutralizing antibodies as VP7. The observations confirm the need for a binary system of rotavirus classification and nomenclature similar to that used for the influenza A viruses; in such a system the neutralization specificity of both VP3 and VP7 would be indicated. PMID:2434522

  9. Synthesis and catalytic water oxidation activities of ruthenium complexes containing neutral ligands.

    PubMed

    Xu, Yunhua; Duan, Lele; Åkermark, Torbjörn; Tong, Lianpeng; Lee, Bao-Lin; Zhang, Rong; Åkermark, Björn; Sun, Licheng

    2011-08-16

    Two dinuclear and one mononuclear ruthenium complexes containing neutral polypyridyl ligands have been synthesised as pre-water oxidation catalysts and characterised by (1)H and (13)C NMR spectroscopy and ESI-MS. Their catalytic water oxidation properties in the presence of [Ce(NH(4))(2)(NO(3))(6)] (Ce(IV)) as oxidant at pH 1.0 have been investigated. At low concentrations of Ce(IV) (5 mM), high turnover numbers of up to 4500 have been achieved. An (18)O-labelling experiment established that both O atoms in the evolved O(2) originate from water. Combined electrochemical study and electrospray ionisation mass spectrometric analysis suggest that ligand exchange between coordinated 4-picoline and free water produces Ru aquo species as the real water oxidation catalysts.

  10. Nucleonic analysis of a preliminary design for the ETF neutral-beam-injector duct shielding

    SciTech Connect

    Urban, W.T.; Seed, T.J.; Dudziak, D.J.

    1980-01-01

    A nucleonic analysis of the Engineering Test Facility Neutral-Beam-Injector duct shielding has been made using a hybrid Monte Carlo/discrete-ordinates method. This method used Monte Carlo to determine internal and external boundary surface sources for a subsequent discrete-ordinates calculation of the neutron and gamma-ray transport through the shield. The analysis also included determination of the energy and angular distribution of neutrons and gamma rays entering the duct from the torus plasma chamber. Confidence in the hybrid method and the results obtained were provided through a comparison with three-dimensional Monte Carlo results.

  11. Lethal, oedema, haemorrhagic activity of spotted butterfish (Scatophagus argus, Linn) sting extract and its neutralization by antiserum and pharmacological antagonists.

    PubMed

    Muhuri, D; Dasgupta, S C; Gomes, A

    2005-06-01

    An attempt has been made in this communication to develop antiserum in rabbit against Scatophagus. argus sting extract. Antiserum did not neutralized the sting extract induced proinflammatory and haemorrhagic activity but successfully neutralized lethality upto 2LD50. Cyproheptadine, indomethacin and BW 755C pretreatment significantly reduced sting extract induced proinflammatory activity. The haemorrhagic activity of sting extract was significantly inhibited by temperature, UV-exposure, EDTA, cyproheptadine, indomethacin and BW 755C pretreatment. The results conclude that the local effects of S.argus venom is likely to be mediated through release of mediators and may be encountered by pharmacological antagonists better than the antiserum.

  12. Neutralizing Antibody and Botulinum Toxin Therapy: A Systematic Review and Meta-analysis.

    PubMed

    Fabbri, Margherita; Leodori, Giorgio; Fernandes, Ricardo M; Bhidayasiri, Roongroj; Marti, Maria Jose; Colosimo, Carlo; Ferreira, Joaquim J

    2016-01-01

    The formation of neutralizing antibodies (NAbs) directed specifically against the active neurotoxin part of the botulinum neurotoxin (BoNT) complex is often cited as a major cause of secondary non-responsiveness (SnR) to treatment. This systematic and meta-analytic review evaluates the frequency of NAbs among patients treated with BoNT therapy for any clinical indication. A comprehensive database search strategy was designed to retrieve relevant clinical data from the published literature up to April 2013. All English-language publications that analyzed NAbs prevalence in more than ten patients were included, regardless of BoNT formulation, assay method, and study design. For the meta-analysis, patients were divided into three categories: secondary nonresponse (SnR) patients, clinically responding patients and all patients, independently of BoNT responsiveness. The meta-analysis included 61 studies reporting data for 8525 patients; 4972 dystonic patients, 1170 patients with spasticity, 294 patients with urologic indications, 396 patient with hyperhidrosis, 1659 patients with glabellar line, and 34 patients with hypersalivation. Among the ‘‘all patients’’ group NAbs frequency was 20%for dystonia, 5.9%for spasticity, and 2.7% for urologic patients and 1.1% for other conditions. The prevalence of NAbs was lower (3.5%) among clinically responding patients and higher in 53.5%SnR patients. About a half of patients with SnR do not have NAbs. NAbs was high among patients treated with RIMA but it was not associated with clinical non-responsiveness. Meta-analysis of the frequency of NAbs and SnR are limited by the heterogeneity of study design and reported outcomes. Indeed the analysis of several factors that can influence the development of NAbs, i.e.,MHCof patients, frequency and site of injection, injection technique, cumulative dose, and toxin denaturation, was not specifically evaluated due to the paucity and heterogeneity of data. The identification of all

  13. Broadly Neutralizing Activity of Zika Virus-Immune Sera Identifies a Single Viral Serotype.

    PubMed

    Dowd, Kimberly A; DeMaso, Christina R; Pelc, Rebecca S; Speer, Scott D; Smith, Alexander R Y; Goo, Leslie; Platt, Derek J; Mascola, John R; Graham, Barney S; Mulligan, Mark J; Diamond, Michael S; Ledgerwood, Julie E; Pierson, Theodore C

    2016-08-01

    Recent epidemics of Zika virus (ZIKV) have been associated with congenital malformation during pregnancy and Guillain-Barré syndrome. There are two ZIKV lineages (African and Asian) that share >95% amino acid identity. Little is known regarding the ability of neutralizing antibodies elicited against one lineage to protect against the other. We investigated the breadth of the neutralizing antibody response following ZIKV infection by measuring the sensitivity of six ZIKV strains to neutralization by ZIKV-confirmed convalescent human serum or plasma samples. Contemporary Asian and early African ZIKV strains were similarly sensitive to neutralization regardless of the cellular source of virus. Furthermore, mouse immune serum generated after infection with African or Asian ZIKV strains was capable of neutralizing homologous and heterologous ZIKV strains equivalently. Because our study only defines a single ZIKV serotype, vaccine candidates eliciting robust neutralizing antibody responses should inhibit infection of both ZIKV lineages, including strains circulating in the Americas. PMID:27481466

  14. Broadly Neutralizing Activity of Zika Virus-Immune Sera Identifies a Single Viral Serotype.

    PubMed

    Dowd, Kimberly A; DeMaso, Christina R; Pelc, Rebecca S; Speer, Scott D; Smith, Alexander R Y; Goo, Leslie; Platt, Derek J; Mascola, John R; Graham, Barney S; Mulligan, Mark J; Diamond, Michael S; Ledgerwood, Julie E; Pierson, Theodore C

    2016-08-01

    Recent epidemics of Zika virus (ZIKV) have been associated with congenital malformation during pregnancy and Guillain-Barré syndrome. There are two ZIKV lineages (African and Asian) that share >95% amino acid identity. Little is known regarding the ability of neutralizing antibodies elicited against one lineage to protect against the other. We investigated the breadth of the neutralizing antibody response following ZIKV infection by measuring the sensitivity of six ZIKV strains to neutralization by ZIKV-confirmed convalescent human serum or plasma samples. Contemporary Asian and early African ZIKV strains were similarly sensitive to neutralization regardless of the cellular source of virus. Furthermore, mouse immune serum generated after infection with African or Asian ZIKV strains was capable of neutralizing homologous and heterologous ZIKV strains equivalently. Because our study only defines a single ZIKV serotype, vaccine candidates eliciting robust neutralizing antibody responses should inhibit infection of both ZIKV lineages, including strains circulating in the Americas.

  15. development of a neutral mass spectrometer dedicated to the analysis of planetary envelopes (NIMEIS)

    NASA Astrophysics Data System (ADS)

    Becker, J.

    2012-12-01

    LATMOS worked for several years on a newly designed instrument suitable for measuring neutral environments as rarefied exosphere of Mars, Venus, Europa, asteroids or Titan for example. This instrument NIMEIS for Neutral and Ion Mass and Energy Imaging Spectrometer has as main features the ability to measure low densities of neutral but also conduct an analysis of mass and energy in an energy range covering the thermal and suprathermal between ~ 1 eV and 20 eV far unexplored. My thesis is divided into two independent parts. First, we optimize the ionization source, that is an innovative concept, and secondly we design the optics of the instrument based on an electrostatic optimization. The ionization source is based on the use of carbon nanotubes and to extract the electrons and ionize the neutral. Employing this technology we can significantly reduce the power, because previously we were using heated filaments. We develop this technology in close collaboration with a laboratory Ajou University (South Korea) that provides us with carbon nanotubes. I did a simulation study of the mode of extraction of electrons from initial tests, from an assembly developed by our laboratory in South Korea. The instrument has been optimized so that the impact on the detector gives us the one hand the energy of the particle and on the other hand the mass of the particle simultaneously and in continuous time. I developed the optics of the instrument using an electrostatic optical software. A comprehensive numerical model has been defined and a prototype is being manufactured.

  16. Dengue Virus Infection-Enhancing Activity in Serum Samples with Neutralizing Activity as Determined by Using FcγR-Expressing Cells

    PubMed Central

    Moi, Meng Ling; Lim, Chang-Kweng; Chua, Kaw Bing; Takasaki, Tomohiko; Kurane, Ichiro

    2012-01-01

    Background Progress in dengue vaccine development has been hampered by limited understanding of protective immunity against dengue virus infection. Conventional neutralizing antibody titration assays that use FcγR-negative cells do not consider possible infection-enhancement activity. We reasoned that as FcγR-expressing cells are the major target cells of dengue virus, neutralizing antibody titration assays using FcγR-expressing cells that determine the sum of neutralizing and infection-enhancing activity, may better reflect the biological properties of antibodies in vivo. Methods and Findings We evaluated serum samples from 80 residents of a dengue endemic country, Malaysia, for neutralizing activity, and infection-enhancing activity at 1∶10 serum dilution by using FcγR-negative BHK cells and FcγR-expressing BHK cells. The serum samples consisted of a panel of patients with acute DENV infection (31%, 25/80) and a panel of donors without acute DENV infection (69%, 55/80). A high proportion of the tested serum samples (75%, 60/80) demonstrated DENV neutralizing activity (PRNT50≥10) and infection-enhancing activity. Eleven of 18 serum samples from patients with acute secondary DENV infection demonstrated neutralizing activity to the infecting serotype determined by using FcγR-negative BHK cells (PRNT50≥10), but not when determined by using FcγR-expressing cells. Conclusion Human serum samples with low neutralizing activity determined by using FcγR-negative cells showed DENV infection-enhancing activity using FcγR-expressing cells, whereas those with high neutralizing activity determined by using FcγR-negative cells demonstrate low or no infection-enhancing activity using FcγR-expressing cells. The results suggest an inverse relationship between neutralizing antibody titer and infection-enhancing activity, and that neutralizing activity determined by using FcγR-expressing cells, and not the activity determined by using FcγR-negative cells, may

  17. Broadly neutralizing DNA vaccine with specific mutation alters the antigenicity and sugar-binding activities of influenza hemagglutinin

    PubMed Central

    Chen, Ming-Wei; Liao, Hsin-Yu; Huang, Yaoxing; Jan, Jia-Tsrong; Huang, Chih-Cheng; Ren, Chien-Tai; Wu, Chung-Yi; Cheng, Ting-Jen Rachel; Ho, David D.; Wong, Chi-Huey

    2011-01-01

    The rapid genetic drift of influenza virus hemagglutinin is an obstacle to vaccine efficacy. Previously, we found that the consensus hemagglutinin DNA vaccine (pCHA5) can only elicit moderate neutralization activities toward the H5N1 clade 2.1 and clade 2.3 viruses. Two approaches were thus taken to improve the protection broadness of CHA5. The first one was to include certain surface amino acids that are characteristic of clade 2.3 viruses to improve the protection profiles. When we immunized mice with CHA5 harboring individual mutations, the antibodies elicited by CHA5 containing P157S elicited higher neutralizing activity against the clade 2.3 viruses. Likewise, the viruses pseudotyped with hemagglutinin containing 157S became more susceptible to neutralization. The second approach was to update the consensus sequence with more recent H5N1 strains, generating a second-generation DNA vaccine pCHA5II. We showed that pCHA5II was able to elicit higher cross-neutralization activities against all H5N1 viruses. Comparison of the neutralization profiles of CHA5 and CHA5II, and the animal challenge studies, revealed that CHA5II induced the broadest protection profile. We concluded that CHA5II combined with electroporation delivery is a promising strategy to induce antibodies with broad cross-reactivities against divergent H5N1 influenza viruses. PMID:21321237

  18. Chromatographic analysis of the neutral and phosphatide glyceryl ethers from various biological sources.

    PubMed

    Hack, M H; Helmy, F M

    1975-04-01

    A method of analysis is described which permits the facile evaluation of the neutral glyceryl ether lipids (mild alkaline hydrolysis) and total glyceryl ether lipid (Vitride reduction) thereby allowing an assessment of the phosphatide contribution. The alkyl and alk-enyl glyceryl ethers are chromatographically resolvable on Gelman glass-fiber type SG and Whatman SG-81 and detected by the periodic acid-Schiff and plasmal reaction, respectively. The attending problems and interpretation are discussed with examples from a number of unicellular organisms and animal tissues.

  19. Performance of thermally activated dolomite for the treatment of Ni and Zn in contaminated neutral drainage.

    PubMed

    Calugaru, Iuliana Laura; Neculita, Carmen Mihaela; Genty, Thomas; Bussière, Bruno; Potvin, Robin

    2016-06-01

    Intensive research is ongoing for developing low-cost and highly efficient materials in metal removal from contaminated effluents. The present study evaluated dolomite [CaMg(CO3)2], both raw and modified by thermal activation (charring), for Ni and Zn treatment in contaminated neutral drainage (CND). Batch adsorption testing (equilibrium and kinetics) were conducted at pH 6, to evaluate the performance of initial vs. modified dolomite, and to assess potential mechanisms of metal removal. Charring of dolomite led to a rigid and porous material, mainly consisting of CaCO3 and MgO, which showed a sorption capacity increased sevenfold for Zn and doubled for Ni, relative to the raw material. In addition, Freundlich model best described the sorption of the both metals by dolomite, whereas the Langmuir model best described their sorption on charred dolomite. Plausible mechanisms of metal removal include cation exchange, surface precipitation and sorption processes, with carbonate ions and magnesium oxides acting as active centers. Based on these results, charred dolomite seems a promising option for the efficient treatment of Ni and Zn in CND. PMID:26897574

  20. Optimization, characterization, sulfation and antitumor activity of neutral polysaccharides from the fruit of Borojoa sorbilis cuter.

    PubMed

    Xu, Fangfang; Liao, Kangsheng; Wu, Yunshan; Pan, Qi; Wu, Lilan; Jiao, Hong; Guo, Dean; Li, Ben; Liu, Bo

    2016-10-20

    Extraction optimization, purification, characterization, sulfation and antitumor activity of polysaccharides from the fruit body of Borojoa sorbilis cuter were investigated in present study. The optimal Ultrahigh Pressure extraction condition was determined as: extraction once with the solid-liquid ratio of 1:10 in 30°C and 1500Mpa for crude polysaccharide (BP) and experimental yield was 8.28%. Four water-soluble polysaccharides named as BP1-1, BP1-2, BP1-3 and BP1-4, with molecular weight of 35.8, 32.4, 30.1 and 27.7kDa, were purified by DEAE Sepharose and Superdex 200 chromatography. On the basis of chemical and spectroscopic analyses, BP1-1-BP1-4 were found to be neutral β-d-galactan containing a (1→4)-linked backbone. S-BP1s with the DSS of 1.18, was sulfated by chloro-sulfonic acid-pyridine method. Furthermore, S-BP1s exhibited significant in vitro antitumor activity against liver cancer HepG2 and lung cancer A549 cells in a dose-dependent manner. The results indicated that S-BP1s could be potentially developed as functional antitumor drug. PMID:27474578

  1. HIV-1 Dual Infected LTNP-EC Patients Developed an Unexpected Antibody Cross-Neutralizing Activity

    PubMed Central

    Pernas, Maria; Sanchez-Merino, Victor; Casado, Concepcion; Merino-Mansilla, Alberto; Olivares, Isabel; Yuste, Eloisa; Lopez-Galindez, Cecilio

    2015-01-01

    This study evaluated the neutralization breadth in dually infected (DI) HIV-1 long-term non-progressor elite controller patients (LTNP-EC) using a representative minipanel of 6 viruses from 5 different subtypes. Our results showed an improved neutralization breadth in DI LTNP-EC patients when compared with matched LTNP single-infected patients. The role of viral diversity in neutralization was estimated with the Shannon Entropy and the p-distance in viral quasispecies. We found a positive correlation between neutralization breadth and diversity within the viral quasispecies. This correlation could explain why a group of LTNP-EC patients developed a broad neutralizing response despite having undetectable levels of viremia. PMID:26258485

  2. HIV-1 Dual Infected LTNP-EC Patients Developed an Unexpected Antibody Cross-Neutralizing Activity.

    PubMed

    Pernas, Maria; Sanchez-Merino, Victor; Casado, Concepcion; Merino-Mansilla, Alberto; Olivares, Isabel; Yuste, Eloisa; Lopez-Galindez, Cecilio

    2015-01-01

    This study evaluated the neutralization breadth in dually infected (DI) HIV-1 long-term non-progressor elite controller patients (LTNP-EC) using a representative minipanel of 6 viruses from 5 different subtypes. Our results showed an improved neutralization breadth in DI LTNP-EC patients when compared with matched LTNP single-infected patients. The role of viral diversity in neutralization was estimated with the Shannon Entropy and the p-distance in viral quasispecies. We found a positive correlation between neutralization breadth and diversity within the viral quasispecies. This correlation could explain why a group of LTNP-EC patients developed a broad neutralizing response despite having undetectable levels of viremia. PMID:26258485

  3. The propeptide is not required to produce catalytically active neutral protease from Bacillus stearothermophilus.

    PubMed

    Mansfeld, Johanna; Petermann, Eva; Dürrschmidt, Peter; Ulbrich-Hofmann, Renate

    2005-02-01

    The thermolysin-like neutral protease from Bacillus stearothermophilus (TLP-ste) is usually produced extracellularly in Bacillus subtilis, where it is expressed as preproenzyme and subsequently processed in an autocatalytic, intramolecular process. To create the basis for the production of inactive mutants of TLP-ste, which cannot be processed in B. subtilis, we studied the expression of TLP-ste and its propeptide in cis and in trans in Escherichia coli. In contrast to thermolysin, subtilisin and alpha-lytic protease, which could be obtained only in the presence of the corresponding propeptides, TLP-ste could be produced as an active mature enzyme in E. coli in the absence of its prosequence. Surprisingly, however, a much more effective access to active mature protease was found when TLP-ste (devoid of its prosequence) was expressed as protein with an N-terminal His6 tag which accumulated in the form of inclusion bodies. Completely unexpected, the protein could be renatured from the inclusion bodies after solubilization in guanidine hydrochloride solutions in high yields. Purification to homogeneity was possible by affinity chromatography on Bacitracin silica as well as by immobilized metal ion affinity chromatography. By addition of separately expressed propeptide to the renaturation mixture yields of renaturation could not be increased significantly, confirming that the propeptide is not essential for proper folding of the enzyme or its stabilization during the folding process. Also in vivo, the expression levels of active mature TLP-ste in Escherichia coli did not significantly differ when the mature sequence was expressed alone or coexpressed with the prosequence in cis or in trans.

  4. Reduction Impairs the Antibacterial Activity but Benefits the LPS Neutralization Ability of Human Enteric Defensin 5

    PubMed Central

    Wang, Cheng; Shen, Mingqiang; Zhang, Naixin; Wang, Song; Xu, Yang; Chen, Shilei; Chen, Fang; Yang, Ke; He, Ting; Wang, Aiping; Su, Yongping; Cheng, Tianmin; Zhao, Jinghong; Wang, Junping

    2016-01-01

    Oxidized human defensin 5 (HD5OX), a Paneth cell-secreted antibacterial peptide with three characteristic disulfide bonds, protects the host from invasion by morbigenous microbes in the small intestine. HD5OX can be reduced by thioredoxin (Trx) in vitro, while the biochemical properties of the reduced linear peptide, HD5RED, remain unclear. Here, we first confirm that HD5RED does exist in vivo. Furthermore, we reveal that the recruitment of HD5RED to the outer membrane of Gram-negative bacteria and to the anionic lipid A is lower than that of HD5OX, and HD5RED is less efficient in penetrating bacterial outer and inner membranes and inducing membrane depolarization, which confers an attenuated antibacterial activity to HD5RED. However, due to its higher structural flexibility, the binding of HD5RED to bacterial lipopolysaccharide (LPS) is markedly stronger than that of HD5OX. Consequently, HD5RED is more effective in suppressing the production of the pro-inflammatory cytokine TNF-α in LPS-stimulated macrophages by blocking the interaction between LPS and LPS-binding protein, thus suggesting that HD5RED might act as a scavenger to neutralize LPS in the gut. This study provides insights into the antibacterial and immunoregulatory effects of HD5RED and expands the known repertoire of the enteric defensins. PMID:26960718

  5. Activation of neutral sphingomyelinase is involved in acute hypoxic pulmonary vasoconstriction

    PubMed Central

    Cogolludo, Angel; Moreno, Laura; Frazziano, Giovanna; Moral-Sanz, Javier; Menendez, Carmen; Castañeda, Javier; González, Constancio; Villamor, Eduardo; Perez-Vizcaino, Francisco

    2009-01-01

    Aims The mechanisms involved in hypoxic pulmonary vasoconstriction (HPV) are not yet fully defined. The aim of the study was to determine the role of protein kinase C ζ (PKCζ) and neutral sphingomyelinase (nSMase) in HPV. Methods and results Ceramide content was measured by immunocytochemistry and voltage-gated potassium channel (KV) currents were recorded by the patch clamp technique in isolated rat pulmonary artery smooth muscle cells (PASMC). Contractile responses were analysed in rat pulmonary arteries mounted in a wire myograph. Pulmonary pressure was recorded in anesthetized open-chest rats. Protein and mRNA expression were measured by western blot and RT–PCR, respectively. We found that hypoxia increased ceramide content in PASMC which was abrogated by inhibition of nSMase, but not acid sphingomyelinase (aSMase). The hypoxia-induced vasoconstrictor response in isolated pulmonary arteries and the inhibition of KV currents were strongly reduced by inhibition of PKCζ or nSMase but not aSMase. The nSMase inhibitor GW4869 prevented HPV in vivo. The vasoconstrictor response to hypoxia was mimicked by exogenous addition of bacterial Smase and ceramide. nSMase2 mRNA expression was ∼10-fold higher in pulmonary compared with mesenteric arteries. In mesenteric arteries, hypoxia failed to increase ceramide but exogenous SMase induced a contractile response. Conclusion nSMase-derived ceramide production and the activation of PKCζ are early and necessary events in the signalling cascade of acute HPV. PMID:19088082

  6. Reduction Impairs the Antibacterial Activity but Benefits the LPS Neutralization Ability of Human Enteric Defensin 5.

    PubMed

    Wang, Cheng; Shen, Mingqiang; Zhang, Naixin; Wang, Song; Xu, Yang; Chen, Shilei; Chen, Fang; Yang, Ke; He, Ting; Wang, Aiping; Su, Yongping; Cheng, Tianmin; Zhao, Jinghong; Wang, Junping

    2016-01-01

    Oxidized human defensin 5 (HD5OX), a Paneth cell-secreted antibacterial peptide with three characteristic disulfide bonds, protects the host from invasion by morbigenous microbes in the small intestine. HD5OX can be reduced by thioredoxin (Trx) in vitro, while the biochemical properties of the reduced linear peptide, HD5RED, remain unclear. Here, we first confirm that HD5RED does exist in vivo. Furthermore, we reveal that the recruitment of HD5RED to the outer membrane of Gram-negative bacteria and to the anionic lipid A is lower than that of HD5OX, and HD5RED is less efficient in penetrating bacterial outer and inner membranes and inducing membrane depolarization, which confers an attenuated antibacterial activity to HD5RED. However, due to its higher structural flexibility, the binding of HD5RED to bacterial lipopolysaccharide (LPS) is markedly stronger than that of HD5OX. Consequently, HD5RED is more effective in suppressing the production of the pro-inflammatory cytokine TNF-α in LPS-stimulated macrophages by blocking the interaction between LPS and LPS-binding protein, thus suggesting that HD5RED might act as a scavenger to neutralize LPS in the gut. This study provides insights into the antibacterial and immunoregulatory effects of HD5RED and expands the known repertoire of the enteric defensins. PMID:26960718

  7. Synthesis, structural characterisation and antibacterial activity of Ag+-doped fluorapatite nanomaterials prepared by neutralization method

    NASA Astrophysics Data System (ADS)

    Stanić, Vojislav; Radosavljević-Mihajlović, Ana S.; Živković-Radovanović, Vukosava; Nastasijević, Branislav; Marinović-Cincović, Milena; Marković, Jelena P.; Budimir, Milica D.

    2015-05-01

    Silver doped fluorapatite nanopowders were synthesised by neutralization method, which consists of dissolving Ag2O in solution of HF and H3PO4 and addition to suspension of Ca(OH)2. The powder XRD, SEM and FTIR studies indicated the formation of a fluorapatite nanomaterials with average length of the particles is about 80 nm and a width of about 15 nm. The FTIR studies show that carbonate content in samples is very small and carbonte ions substitute both phosphate and hydroxyl groups in the crystal structure of samples, forming AB-type fluorapatite. Antibacterial studies have demonstrated that all Ag+-doped fluorapatite samples exhibit bactericidal effect against pathogens: Staphylococcus aureus, Micrococcus luteus and Kllebsiela pneumoniae. Antibacterial activity increased with the increase of Ag+ in the samples. The atomic force microscopy studies revealed extensive damage to the bacterial cell envelops in the presence of Ag+-doped fluorapatite particles which may lead to their death. The synthesized Ag+-doped fluorapatite nanomaterials are promising as antibacterial biomaterials in orthopedics and dentistry.

  8. Utilization of activated CO2-neutralized red mud for removal of arsenate from aqueous solutions.

    PubMed

    Sahu, Ramesh Chandra; Patel, Rajkishore; Ray, Bankim Chandra

    2010-07-15

    A laboratory study was conducted to investigate the ability of activated CO(2)-neutralized red mud (ANRM) for the removal of arsenate from the aqueous solutions. The batch adsorption experiments were conducted with respect to adsorbent dose, equilibrium pH, contact time, initial arsenate concentration, kinetics, Langmuir isotherms. The mechanisms involved in adsorption of arsenate ions on ANRM were characterized by using XRD, FT-IR, UV-vis, SEM/EDX, and chemical methods. The percentage removal was found to increase gradually with decrease of pH and maximum removal was achieved at pH approximately 4. Adsorption kinetic studies revealed that the adsorption process followed pseudo-second-order kinetics and equilibrates within 24 h. FT-IR spectra of ANRM before and after adsorption reveals the binding of arsenate to the adsorbent. The adsorption data were fitted to linearly transformed Langmuir isotherm with R(2) (correlation coefficient)>0.99. Arsenate adsorbed ANRM can be regenerated using NaOH solution at pH 12.0.

  9. Consistent analysis of neutral- and charged-current (anti)neutrino scattering off carbon

    SciTech Connect

    Ankowski, Artur M.

    2015-05-15

    Good understanding of the cross sections for (anti)neutrino scattering off nuclear targets in the few-GeV energy region is a prerequisite for the correct interpretation of results of ongoing and planned oscillation experiments. To clarify a possible source of disagreement between recent measurements of the cross sections on carbon, we analyze the available data within an approach based on the realistic spectral function of carbon, treating neutral-current elastic (NCE) and charged-current quasielastic (CCQE) processes on equal footing. We show that the axial mass from the shape analysis of the MiniBooNE data is in good agreement with the results reported by the BNL E734 and NOMAD Collaborations. However, the combined analysis of the NCE and CCQE data does not seem to support the contribution of multinucleon final states being large enough to explain the normalization of the MiniBooNE-reported cross sections.

  10. Consistent analysis of neutral- and charged-current (anti)neutrino scattering off carbon

    NASA Astrophysics Data System (ADS)

    Ankowski, Artur M.

    2015-05-01

    Good understanding of the cross sections for (anti)neutrino scattering off nuclear targets in the few-GeV energy region is a prerequisite for the correct interpretation of results of ongoing and planned oscillation experiments. To clarify a possible source of disagreement between recent measurements of the cross sections on carbon, we analyze the available data within an approach based on the realistic spectral function of carbon, treating neutral-current elastic (NCE) and charged-current quasielastic (CCQE) processes on equal footing. We show that the axial mass from the shape analysis of the MiniBooNE data is in good agreement with the results reported by the BNL E734 and NOMAD Collaborations. However, the combined analysis of the NCE and CCQE data does not seem to support the contribution of multinucleon final states being large enough to explain the normalization of the MiniBooNE-reported cross sections.

  11. Mean square delay dependent-probability-distribution stability analysis of neutral type stochastic neural networks.

    PubMed

    Muralisankar, S; Manivannan, A; Balasubramaniam, P

    2015-09-01

    The aim of this manuscript is to investigate the mean square delay dependent-probability-distribution stability analysis of neutral type stochastic neural networks with time-delays. The time-delays are assumed to be interval time-varying and randomly occurring. Based on the new Lyapunov-Krasovskii functional and stochastic analysis approach, a novel sufficient condition is obtained in the form of linear matrix inequality such that the delayed stochastic neural networks are globally robustly asymptotically stable in the mean-square sense for all admissible uncertainties. Finally, the derived theoretical results are validated through numerical examples in which maximum allowable upper bounds are calculated for different lower bounds of time-delay.

  12. Inhibition of toxic activities of Bothrops asper venom and other crotalid snake venoms by a novel neutralizing mixture.

    PubMed

    Borkow, G; Gutierrez, J M; Ovadia, M

    1997-12-01

    The majority of snake bites in Central America are caused by Bothrops asper, whose venom induce complex local effects such as myonecrosis, edema and especially hemorrhage. These effects are only partially neutralized by the clinically used antivenom, even when administered rapidly after envenomation. Recently we screened 49 substances for antihemorrhagic activity and found that a mixture composed of CaNa2, EDTA, a B. asper serum fraction (natural antidote), and the currently used horse polyvalent antiserum is highly effective in the neutralization of local and systemic hemorrhage developing after B. asper envenomation (Borkow et al., Toxicon 35, 865-877, 1997). In the present study we screened the best six antihemorrhagic compounds for their capacity to neutralize the lethal activity in mice and the proteolytic, hemolytic, and antiattachment activities in vitro of the venom. The compounds tested included the currently used horse antivenom, rabbit antiserum against whole B. asper venom or against heated venom, B. asper and Natrix tessellata serum fractions, and CaNa2 EDTA. The constituents of the antihemorrhagic mixture were also the best inhibitors of the other examined toxic activities. Importantly, the mixture effectively neutralized toxic activities of an additional nine venoms from snakes abundant in Central America. This work suggests that the polyvalent antivenom used in Central America could be enriched with a B. asper serum fraction producing a more effective antivenom. In addition, the local application of CaNa2 EDTA to neutralize hemorrhagic toxins, immediately after a snake bite, may provide rapid inhibition of local damage caused by the venoms. PMID:9439739

  13. Relationships between adaptive and neutral genetic diversity and ecological structure and functioning: a meta-analysis.

    PubMed

    Whitlock, Raj

    2014-07-01

    Understanding the effects of intraspecific genetic diversity on the structure and functioning of ecological communities is a fundamentally important part of evolutionary ecology and may also have conservation relevance in identifying the situations in which genetic diversity coincides with species-level diversity.Early studies within this field documented positive relationships between genetic diversity and ecological structure, but recent studies have challenged these findings. Conceptual synthesis has been hampered because studies have used different measures of intraspecific variation (phenotypically adaptive vs. neutral) and have considered different measures of ecological structure in different ecological and spatial contexts. The aim of this study is to strengthen conceptual understanding by providing an empirical synthesis quantifying the relationship between genetic diversity and ecological structure.Here, I present a meta-analysis of the relationship between genetic diversity within plant populations and the structure and functioning of associated ecological communities (including 423 effect sizes from 70 studies). I used Bayesian meta-analyses to examine (i) the strength and direction of this relationship, (ii) the extent to which phenotypically adaptive and neutral (molecular) measures of diversity differ in their association with ecological structure and (iii) variation in outcomes among different measures of ecological structure and in different ecological contexts.Effect sizes measuring the relationship between adaptive diversity (genotypic richness) and both community- and ecosystem-level ecological responses were small, but significantly positive. These associations were supported by genetic effects on species richness and productivity, respectively.There was no overall association between neutral genetic diversity and measures of ecological structure, but a positive correlation was observed under a limited set of demographic conditions. These

  14. Relationships between adaptive and neutral genetic diversity and ecological structure and functioning: a meta-analysis

    PubMed Central

    Whitlock, Raj

    2014-01-01

    Understanding the effects of intraspecific genetic diversity on the structure and functioning of ecological communities is a fundamentally important part of evolutionary ecology and may also have conservation relevance in identifying the situations in which genetic diversity coincides with species-level diversity.Early studies within this field documented positive relationships between genetic diversity and ecological structure, but recent studies have challenged these findings. Conceptual synthesis has been hampered because studies have used different measures of intraspecific variation (phenotypically adaptive vs. neutral) and have considered different measures of ecological structure in different ecological and spatial contexts. The aim of this study is to strengthen conceptual understanding by providing an empirical synthesis quantifying the relationship between genetic diversity and ecological structure.Here, I present a meta-analysis of the relationship between genetic diversity within plant populations and the structure and functioning of associated ecological communities (including 423 effect sizes from 70 studies). I used Bayesian meta-analyses to examine (i) the strength and direction of this relationship, (ii) the extent to which phenotypically adaptive and neutral (molecular) measures of diversity differ in their association with ecological structure and (iii) variation in outcomes among different measures of ecological structure and in different ecological contexts.Effect sizes measuring the relationship between adaptive diversity (genotypic richness) and both community- and ecosystem-level ecological responses were small, but significantly positive. These associations were supported by genetic effects on species richness and productivity, respectively.There was no overall association between neutral genetic diversity and measures of ecological structure, but a positive correlation was observed under a limited set of demographic conditions. These

  15. Important increase of negative secondary ion sensitivity during SIMS analysis by neutral cesium deposition

    NASA Astrophysics Data System (ADS)

    Philipp, P.; Wirtz, T.; Migeon, H.-N.; Scherrer, H.

    2006-07-01

    The Cation Mass Spectrometer (CMS) is a SIMS prototype developed in our laboratory in order to perform quantitative analysis with optimal sensitivity and high depth and lateral resolution in the MCs x+ and M - modes. For this aim, a patented neutral Cs evaporator for varying the Cs surface concentration over the whole range has been developed. The sputtering and the Cs introduction have been decoupled successfully by applying simultaneously X y+ bombardment and neutral Cs deposition. X stands for any element except Cs. Currently the CMS is equipped with a Cs + and a Ga + gun. These guns are used for studying the useful yield variations of negative secondary ions (Si -, Al -, Ni -, In -, P -) with respect to the Cs surface concentration. Furthermore, the observed variations of the useful yield were linked to work function shifts. Qualitative agreement with the predictions of the electron tunnelling model are obtained. Several applications give further evidence of successful optimization of the useful yield for elements with high electron affinity.

  16. ACTIVE IMMUNITY PRODUCED BY SO CALLED BALANCED OR NEUTRAL MIXTURES OF DIPHTHERIA TOXIN AND ANTITOXIN.

    PubMed

    Smith, T

    1909-03-01

    The foregoing and earlier data taken together demonstrate that an active immunity lasting several years can be produced in guinea-pigs, by the injection of toxin-antitoxin mixtures which have no recognizable harmful effect either immediate or remote. They also show, what might have been anticipated, that under the same conditions mixtures which produce local lesions and which, therefore, contain an excess of toxin produce a much higher degree of immunity than the neutral mixtures, and that an excess of antitoxin reduces the possibility of producing an active immunity, and may extinguish it altogether. There is, therefore, a certain definite relation between the components of the mixture and the degree of immunity producible. Furthermore, toxin-antitoxin mixtures do not change materially within five days at room temperature. They are apparently more efficacious at the end of forty-eight hours than immediately after preparation. The experiments finally prove that a relatively high degree of active immunity can be induced by a harmless procedure, whereas the use of toxin alone leading to very severe local lesions is incapable of producing more than an insignificant protection. The method, therefore, invites further tests in regard to its ultimate applicability to the human being. Unless the subcutis of the guinea-pig reacts to toxin-antitoxin mixtures in a manner peculiar to itself, a practical, easily controlled method for active immunization can be worked out which should afford a larger protection than the serum alone and avoid the complications associated with horse serum. That proportion of toxin and antitoxin which would produce the highest desirable immunity consistent with the least discomfort would have to be carefully worked out for the human subject. From the nature of the immunity induced it is obvious, however, that such a method of immunization cannot take the place of a large dose of antitoxin in exposed individuals who must be protected at once. It

  17. Structural characterization of a novel neutral polysaccharide from Lentinus giganteus and its antitumor activity through inducing apoptosis.

    PubMed

    Tian, Yuting; Zhao, Yingting; Zeng, Hongliang; Zhang, Yaling; Zheng, Baodong

    2016-12-10

    A novel neutral polysaccharide (LGPS-1), with a molecular weight of 1.547×10(5)Da, was isolated from Lentinus giganteus by precipitation and purification. The monosaccharides included d-mannose (Man), d-glucose (Glc) and d-galactose (Gal) with a molar ratio of 3.0:4.1:7.1. The backbone of LGPS-1 was composed of 1,6-Galp and 1,3,6-Manp whereas the branches were composed of 1,6-Glcp and 1-Glcp. The anticancer efficacy of LGPS-1 was assessed using HepG2 hepatocellular carcinoma cells. The results showed that LGPS-1 inhibited the proliferation of HepG2 cells and also induced the activation of caspase-3, and cleavage of PARP-1. Western blot analysis revealed that LGSP-1 significantly induced a loss of mitochondrial membrane potential (Δym), increased the ratio of Bax/Bcl-2, promoted the release of cytochrome c into cytoplasm as well as inhibited the phosphorylation of Akt in HepG2 cells. These findings suggest that LGPS-1 induced apoptosis in HepG2 cells through intrinsic mitochondrial apoptosis and PI3K/Akt signaling pathways. PMID:27577914

  18. Neutralization Assay for Chikungunya Virus Infection: Plaque Reduction Neutralization Test.

    PubMed

    Azami, Nor Azila Muhammad; Moi, Meng Ling; Takasaki, Tomohiko

    2016-01-01

    Neutralization assay is a technique that detects and quantifies neutralizing antibody in serum samples by calculating the percentage of reduction of virus activity, as the concentration of virus used is usually constant. Neutralizing antibody titer is conventionally determined by calculating the percentage reduction in total virus infectivity by counting and comparing number of plaques (localized area of infection due to cytopathic effect) with a standard amount of virus. Conventional neutralizing test uses plaque-reduction neutralization test (PRNT) to determine neutralizing antibody titers against Chikungunya virus (CHIKV). Here we describe the plaque reduction neutralization assay (PRNT) using Vero cell lines to obtain neutralizing antibody titers.

  19. NON-NEUTRALIZED ELECTRIC CURRENT PATTERNS IN SOLAR ACTIVE REGIONS: ORIGIN OF THE SHEAR-GENERATING LORENTZ FORCE

    SciTech Connect

    Georgoulis, Manolis K.; Titov, Viacheslav S.; Mikic, Zoran

    2012-12-10

    Using solar vector magnetograms of the highest available spatial resolution and signal-to-noise ratio, we perform a detailed study of electric current patterns in two solar active regions (ARs): a flaring/eruptive and a flare-quiet one. We aim to determine whether ARs inject non-neutralized (net) electric currents in the solar atmosphere, responding to a debate initiated nearly two decades ago that remains inconclusive. We find that well-formed, intense magnetic polarity inversion lines (PILs) within ARs are the only photospheric magnetic structures that support significant net current. More intense PILs seem to imply stronger non-neutralized current patterns per polarity. This finding revises previous works that claim frequent injections of intense non-neutralized currents by most ARs appearing in the solar disk but also works that altogether rule out injection of non-neutralized currents. In agreement with previous studies, we also find that magnetically isolated ARs remain globally current-balanced. In addition, we confirm and quantify the preference of a given magnetic polarity to follow a given sense of electric currents, indicating a dominant sense of twist in ARs. This coherence effect is more pronounced in more compact ARs with stronger PILs and must be of sub-photospheric origin. Our results yield a natural explanation of the Lorentz force, invariably generating velocity and magnetic shear along strong PILs, thus setting a physical context for the observed pre-eruption evolution in solar ARs.

  20. Electric fields measured by ISEE-1 within and near the neutral sheet during quiet and active times

    NASA Technical Reports Server (NTRS)

    Cattell, C. A.; Mozer, F. S.

    1982-01-01

    An understanding of the physical processes occurring in the magnetotail and plasmasheet during different interplanetary magnetic field orientations and differing levels of ground magnetic activity is crucial for the development of a theory of energy transfer from the solar wind to the particles which produce auroral arcs. In the present investigation, the first observations of electric fields during neutral sheet crossings are presented, taking into account the statistical correlations of the interplanetary magnetic field direction and ground activity with the character of the electric field. The electric field data used in the study were obtained from a double probe experiment on the ISEE-1 satellite. The observations suggest that turbulent electric and magnetic fields are intimately related to plasma acceleration in the neutral sheet and to the processes which create auroral particles.

  1. Characterization of a Family GH5 Xylanase with Activity on Neutral Oligosaccharides and Evaluation as a Pulp Bleaching Aid ▿

    PubMed Central

    Gallardo, Óscar; Fernández-Fernández, María; Valls, Cristina; Valenzuela, Susana Valeria; Roncero, M. Blanca; Vidal, Teresa; Díaz, Pilar; Pastor, F. I. Javier

    2010-01-01

    A new bacterial xylanase belonging to family 5 of glycosyl hydrolases was identified and characterized. The xylanase, Xyn5B from Bacillus sp. strain BP-7, was active on neutral, nonsubstituted xylooligosaccharides, showing a clear difference from other GH5 xylanases characterized to date that show a requirement for methyl-glucuronic acid side chains for catalysis. The enzyme was evaluated on Eucalyptus kraft pulp, showing its effectiveness as a bleaching aid. PMID:20656870

  2. Short communication: Presence of neutral metallopeptidase (npr) gene and proteolytic activity of Bacillus cereus isolated from dairy products.

    PubMed

    Montanhini, M T M; Colombo, M; Nero, L A; Bersot, L S

    2013-09-01

    The control of proteolytic microorganisms is one of the main challenges of the dairy industry, due to their spoilage activity that jeopardizes the quality of their products. Seventy-four Bacillus cereus strains isolated from powdered, UHT, and pasteurized milks were tested for the presence of the neutral metallopeptidase (npr) gene and proteolytic activity at 7, 10, 25, 30, and 37°C. All strains had the npr gene, and proteolytic activity increased with the incubation temperature. The obtained results highlight the relevance of B. cereus as a spoiling agent in the dairy industry in terms of its genetic predisposition for proteolytic capacity, especially at room temperature.

  3. Short communication: Presence of neutral metallopeptidase (npr) gene and proteolytic activity of Bacillus cereus isolated from dairy products.

    PubMed

    Montanhini, M T M; Colombo, M; Nero, L A; Bersot, L S

    2013-09-01

    The control of proteolytic microorganisms is one of the main challenges of the dairy industry, due to their spoilage activity that jeopardizes the quality of their products. Seventy-four Bacillus cereus strains isolated from powdered, UHT, and pasteurized milks were tested for the presence of the neutral metallopeptidase (npr) gene and proteolytic activity at 7, 10, 25, 30, and 37°C. All strains had the npr gene, and proteolytic activity increased with the incubation temperature. The obtained results highlight the relevance of B. cereus as a spoiling agent in the dairy industry in terms of its genetic predisposition for proteolytic capacity, especially at room temperature. PMID:23849645

  4. Between detection and neutralization.

    SciTech Connect

    Snell, Mark Kamerer; Green, Mary Wilson; Adams, Douglas Glenn; Pritchard, Daniel Allison

    2005-08-01

    Security system analytical performance analysis is generally based on the probability of system effectiveness. The probability of effectiveness is a function of the probabilities of interruption and neutralization. Interruption occurs if the response forces are notified in sufficient time to engage the adversary. Neutralization occurs if the adversary attack is defeated after the security forces have actively engaged the adversary. Both depend upon communications of data. This paper explores details of embedded communications functions that are often assumed to be inconsequential. It is the intent of the authors to bring focus to an issue in security system modeling that, if not well understood, has the potential to be a deciding factor in the overall system failure or effectiveness.

  5. Towards energy neutrality by optimising the activated sludge process of the WWTP Bochum-Ölbachtal.

    PubMed

    Marner, S T; Schröter, D; Jardin, N

    2016-01-01

    As a result of Ruhrverband's regularly performed energy audits the wastewater treatment plant (WWTP) Bochum-Ölbachtal shows substantial deficits concerning energy efficiency. Due to the energy consumption of internal recirculation, mixers and return activated sludge (RAS) pumping the existing pre-denitrification process configuration offers a specific energy consumption for biological treatment of 23 kWh (PE · a)(-1). In order to optimise the energy situation and to improve the treatment efficiency, the process layout was changed completely to a three-stage step-feed process. By optimising the hydraulic conditions, it was possible to reconstruct the plant with a free flow throughout the whole biological treatment system without any additional pumping. The total investment costs for this process scheme were 3.9 million €. These costs could be partly offset against the wastewater charge paid (2.9 million €). Compared to the overall energy consumption before the process modification, today the energy consumption for biological treatment amounts to 12.4 kWh (PE · a)(-1). The highest saving potential has been achieved by optimising mixing and reducing the energy demand for internal recirculation and RAS pumping. In the case of the WWTP Bochum-Ölbachtal, the modification of the treatment process not only results in an improved energy situation but also increased the treatment efficiency in such a way that the nitrogen concentration in the effluent could be constantly kept below 5 mg L(-1) N(tot), which provides the basis for being exempted from the wastewater discharge for nitrogen. As a result of all these measures, the rate of self-sufficiency by using biogas from the digester in combined heat and power units has been increased substantially from 60% before process modifications to 97%. With the upcoming optimisation measures, a further increase of self-sufficiency is expected to finally achieve energy neutrality based on yearly averages. The example of the

  6. Towards energy neutrality by optimising the activated sludge process of the WWTP Bochum-Ölbachtal.

    PubMed

    Marner, S T; Schröter, D; Jardin, N

    2016-01-01

    As a result of Ruhrverband's regularly performed energy audits the wastewater treatment plant (WWTP) Bochum-Ölbachtal shows substantial deficits concerning energy efficiency. Due to the energy consumption of internal recirculation, mixers and return activated sludge (RAS) pumping the existing pre-denitrification process configuration offers a specific energy consumption for biological treatment of 23 kWh (PE · a)(-1). In order to optimise the energy situation and to improve the treatment efficiency, the process layout was changed completely to a three-stage step-feed process. By optimising the hydraulic conditions, it was possible to reconstruct the plant with a free flow throughout the whole biological treatment system without any additional pumping. The total investment costs for this process scheme were 3.9 million €. These costs could be partly offset against the wastewater charge paid (2.9 million €). Compared to the overall energy consumption before the process modification, today the energy consumption for biological treatment amounts to 12.4 kWh (PE · a)(-1). The highest saving potential has been achieved by optimising mixing and reducing the energy demand for internal recirculation and RAS pumping. In the case of the WWTP Bochum-Ölbachtal, the modification of the treatment process not only results in an improved energy situation but also increased the treatment efficiency in such a way that the nitrogen concentration in the effluent could be constantly kept below 5 mg L(-1) N(tot), which provides the basis for being exempted from the wastewater discharge for nitrogen. As a result of all these measures, the rate of self-sufficiency by using biogas from the digester in combined heat and power units has been increased substantially from 60% before process modifications to 97%. With the upcoming optimisation measures, a further increase of self-sufficiency is expected to finally achieve energy neutrality based on yearly averages. The example of the

  7. Neutralizer optimization

    NASA Technical Reports Server (NTRS)

    Patterson, Michael J.; Mohajeri, Kayhan

    1991-01-01

    The preliminary results of a test program to optimize a neutralizer design for 30 cm xenon ion thrusters are discussed. The impact of neutralizer geometry, neutralizer axial location, and local magnetic fields on neutralizer performance is discussed. The effect of neutralizer performance on overall thruster performance is quantified, for thruster operation in the 0.5-3.2 kW power range. Additionally, these data are compared to data published for other north-south stationkeeping (NSSK) and primary propulsion xenon ion thruster neutralizers.

  8. Neutral current π0 analysis in the NO νA experiment

    NASA Astrophysics Data System (ADS)

    Sahoo, Himansu

    2015-04-01

    The NO νA experiment is a long-baseline accelerator based neutrino oscillation experiment using an upgraded NuMI neutrino beam from Fermilab. By studying electron neutrino and antineutrino appearance, the experiment aims to determine the neutrino mass hierarchy and will provide constraints on CP violation. NO νA Detector construction is complete and both detectors have started collecting neutrino data. Interactions in which an energetic π0 is produced in the final state are one of the dominant sources of backgrounds to the νe search, but also provide a rich sample of events to characterize the detector. Reconstruction of photon showers from π0 decays provide input for calibration over the full energy range of interest for the νe appearance analysis. Large uncertainties in model prediction requires a precise measurement of NC π0 production rates and cross-sections using NO νA data. In this talk, we will present the neutral current π0 analysis in NO νA and the first results using data from Near Detector.

  9. Analysis of Solar Wind Samples Returned by Genesis Using Laser Post Ionization Secondary Neutral Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Veryovkin, I. V.; Calaway, W. F.; Tripa, C. E.; Pellin, M. J.; Burnett, D. S.

    2005-12-01

    A new secondary neutral mass spectrometry (SNMS) instrument implementing laser post ionization (LPI) of ion sputtered and laser desorbed neutral species has been developed and constructed for the specific purpose of quantitative analysis of metallic elements at ultra trace levels in solar wind collector samples returned to Earth by the Genesis Discovery mission. The first LPI SNMS measurements are focusing on determining Al, Ca, Cr, and Mg in these samples. These measurements provide the first concentration and isotopic abundances determinations for several key metallic elements and also elucidate possible fractionation effects between the photosphere and the solar wind compositions. It is now documented that Genesis samples suffered surface contamination both during flight and during the breach of the Sample Return Capsule when it crashed. Since accurate quantitative analysis is compromised by sample contamination, several features have been built into the new LPI SNMS instrument to mitigate this difficulty. A normally-incident, low-energy (<500 eV) ion beam combined with a keV energy ion beam and a desorbing laser beam (both microfocused) enables dual beam analyses. The low-energy ion beam can be used to remove surface contaminant by sputtering with minimum ion beam mixing. This low-energy beam also will be used to perform ion beam milling, while either the microfocused ion or laser beam probes the solar wind elemental compositions as a function of sample depth. Because of the high depth resolution of dual beam analyses, such depth profiles clearly distinguish between surface contaminants and solar wind implanted atoms. In addition, in-situ optical and electron beam imaging for observing and avoiding particulates and scratches on solar wind sample surfaces is incorporated in the new LPI SNMS instrument to further reduce quantification problems. The current status of instrument tests and analyses will be presented. This work is supported by the U. S. Department of

  10. Mass Spectrometric Collisional Activation and Product Ion Mobility of Human Serum Neutral Lipid Extracts

    PubMed Central

    Hankin, Joseph A.; Barkley, Robert M.; Zemski-Berry, Karin; Deng, Yiming; Murphy, Robert C.

    2016-01-01

    A novel method for lipid analysis called CTS (collisional activation and traveling wave mass spectrometry) involving tandem mass spectrometry of all precursor ions with ion mobility determinations of all product ions was applied to a sample of human serum. The resulting four dimensional data set (precursor ion, product ion, ion mobility values, and intensity) was found to be useful for characterization of lipids as classes as well as identification of specific species. Utilization of ion mobility measurements of the product ions is a novel approach for lipid analysis. The trends and patterns of product mobility values when visually displayed yield information on lipid classes and specific species independent of mass determination. The collection of a comprehensive set of data that incorporates all precursor-product relationships combined with ion mobility measurements of all products enables data analysis where different molecular properties can be juxtaposed and analyzed to assist with class and species identification. Overall, CTS is powerful, specific, and comprehensive method for lipid analysis. PMID:27213895

  11. Laser neutralization

    SciTech Connect

    Peterson, O.G.

    1986-06-17

    Laser photodetachment of the excess electron to neutralize relativistic ions offers many advantages over the more conventional collisional methods using gases or thin foils as the neutralization agents. Probably the two most important advantages of laser photodetachment are the generation of a compact and low divergence beam, and the production of intense neutral beams at very high efficiency (approximately 90%). The high intensities or high current densities of the neutral beam result from the fixed maximum divergence that can be added to the beam by photodetachment of the charge using laser intensity of fixed wavelength and incident angle. The high neutralization efficiency is possible because there is no theoretical maximum to the neutralization efficiency, although higher efficiencies require higher laser powers and, therefore, costs. Additional advantages include focusability of the laser light onto the ion beam to maximize its efficacy. There certainly is no residual gas left in the particle beam path as is typical with gas neutralizers. The photodetachment process leaves the neutral atoms in the ground state so there is no excited state fluorescence to interfere with the subsequent beam sensing. Finally, since the beams to be neutralized are very high powered, for a large range of neutralization efficiencies the neutral beam can be increased more by increasing the power to the laser neutralizer than by adding an equal amount of power to the primary accelerator. 26 figs.

  12. Monoclonal antibody analysis of neutralization and antibody-dependent enhancement of feline infectious peritonitis virus.

    PubMed

    Corapi, W V; Olsen, C W; Scott, F W

    1992-11-01

    Fifty-four monoclonal antibodies (MAbs) to feline infectious peritonitis virus (FIPV) were characterized according to protein specificity, immunoglobulin subclass, virus neutralization, reactivity with different coronaviruses, and ability to induce antibody-dependent enhancement (ADE) of FIPV infection in vitro. The MAbs were found to be specific for one of three structural proteins of FIPV. A total of 47 MAbs were specific for the 205-kDa spike protein (S), 3 MAbs were specific for the 45-kDa nucleocapsid protein (N), and 4 MAbs were specific for the 26- to 28-kDa membrane protein (M). The S-specific MAbs showed various degrees of cross-reactivity with strains of FIPV, feline enteric coronavirus, canine coronavirus, and porcine transmissible gastroenteritis virus. Nineteen S-specific MAbs neutralized FIPV. A total of 15 of the neutralizing MAbs induced ADE, and all but 1 were of the immunoglobulin G2a subclass. The remaining four neutralizing MAbs that did not induce ADE were of the immunoglobulin G1 subclass. Two S-specific MAbs induced ADE but were nonneutralizing. None of the N- or M-specific MAbs was neutralizing or induced ADE. On the basis of the reactivity patterns of the MAbs with FIPV and related coronaviruses, it was concluded that there is a minimum of five neutralizing sites on S. In most instances, neutralizing MAbs were able to induce ADE, demonstrating a direct relationship between neutralization and enhancement. The difference in immunoglobulin subclass between neutralizing MAbs that induced ADE and those that did not induce ADE suggests that there may be a restriction in the immunoglobulin subclasses capable of mediating ADE.

  13. Quasi-linear analysis of ion Weibel instability in the earth's neutral sheet

    NASA Technical Reports Server (NTRS)

    Lui, Anthony T. Y.; Yoon, Peter H.; Chang, Chia-Lie

    1993-01-01

    A quasi-linear analysis of the ion Weibel instability (IWI) for waves with parallel propagation is carried out for parameters appropriate to the earth's neutral sheet during the substorm interval. For ion drift speed reaching sizable fraction of the ion thermal speed, unstable waves grow to a nonlinear regime in a time interval greater than an ion gyroperiod. The saturation level is attained with current density reduced to about 15-28 percent of its preactivity level. The unstable wave amplitude normalized to the initial ambient field is found to be in the range of 0.2-0.8. This is accompanied by ion heating along the magnetic field with the parallel temperature being enhanced by 25-90 percent. Thus, the IWI can provide nonadiabatic heating of ions in current disruptions during substorms. The associated anomalous resistivity is estimated to be about 1 x 10 exp -7 to 1 x 10 exp -6 s, which is about 11 to 12 orders of magnitude above the classical resistivity.

  14. Differential Neutralizing Activities of a Single Domain Camelid Antibody (VHH) Specific for Ricin Toxin’s Binding Subunit (RTB)

    PubMed Central

    Herrera, Cristina; Vance, David J.; Eisele, Leslie E.; Shoemaker, Charles B.; Mantis, Nicholas J.

    2014-01-01

    Ricin, a member of the A-B family of ribosome-inactivating proteins, is classified as a Select Toxin by the Centers for Disease Control and Prevention because of its potential use as a biothreat agent. In an effort to engineer therapeutics for ricin, we recently produced a collection of alpaca-derived, heavy-chain only antibody VH domains (VHH or “nanobody”) specific for ricin’s enzymatic (RTA) and binding (RTB) subunits. We reported that one particular RTB-specific VHH, RTB-B7, when covalently linked via a peptide spacer to different RTA-specific VHHs, resulted in heterodimers like VHH D10/B7 that were capable of passively protecting mice against a lethal dose challenge with ricin. However, RTB-B7 itself, when mixed with ricin at a 1∶10 toxin:antibody ratio did not afford any protection in vivo, even though it had demonstrable toxin-neutralizing activity in vitro. To better define the specific attributes of antibodies associated with ricin neutralization in vitro and in vivo, we undertook a more thorough characterization of RTB-B7. We report that RTB-B7, even at 100-fold molar excess (toxin:antibody) was unable to alter the toxicity of ricin in a mouse model. On the other hand, in two well-established cytotoxicity assays, RTB-B7 neutralized ricin with a 50% inhibitory concentration (IC50) that was equivalent to that of 24B11, a well-characterized and potent RTB-specific murine monoclonal antibody. In fact, RTB-B7 and 24B11 were virtually identical when compared across a series of in vitro assays, including adherence to and neutralization of ricin after the toxin was pre-bound to cell surface receptors. RTB-B7 differed from both 24B11 and VHH D10/B7 in that it was relatively less effective at blocking ricin attachment to receptors on host cells and was not able to form high molecular weight toxin:antibody complexes in solution. Whether either of these activities is important in ricin toxin neutralizing activity in vivo remains to be determined. PMID

  15. Synthesis and characterization of higher amino acid Schiff bases, as monosodium salts and neutral forms. Investigation of the intramolecular hydrogen bonding in all Schiff bases, antibacterial and antifungal activities of neutral forms

    NASA Astrophysics Data System (ADS)

    Güngör, Özlem; Gürkan, Perihan

    2014-09-01

    Schiff bases derived from 5-nitro-salicylaldehyde and 4-aminobutyric acid, 5-aminopentanoic acid and 6-aminohexanoic acid were synthesized both as monosodium salts (1a-3a) and neutral forms (1b-3b). The monosodium-Schiff bases were characterized by elemental analysis, 1H/13C NMR, IR, powder XRD, UV-vis spectra and conductivity measurements. The neutral-Schiff bases were characterized by elemental analysis, 1H/13C NMR, 2D NMR (HMQC), mass, IR, powder XRD, UV-vis spectra and conductivity measurements. The intramolecular hydrogen bonding and related tautomeric equilibria in all the Schiff bases were studied by UV-vis and 1H NMR spectra in solution. Additionally, the neutral-Schiff bases were screened against Staphylococcus aureus-EB18, S. aureus-ATCC 25923, Escherichia coli-ATCC 11230, Candida albicans-M3 and C. albicans-ATCC 16231.

  16. Influences of acidic reaction and hydrolytic conditions on monosaccharide composition analysis of acidic, neutral and basic polysaccharides.

    PubMed

    Wang, Qing-Chi; Zhao, Xia; Pu, Jiang-Hua; Luan, Xiao-Hong

    2016-06-01

    Monosaccharide composition analysis is important for structural characterization of polysaccharides. To investigate the influences of acidic reaction and hydrolytic conditions on monosaccharide composition analysis of polysaccharides, we chose alginate, starch, chitosan and chondroitin sulfate as representative of acidic, neutral, basic and complex polysaccharides to compare the release degree of monosaccharides under different hydrolytic conditions. The hydrolysis stability of 10 monosaccharide standards was also explored. Results showed that the basic sugars were hard to release but stable, the acidic sugars (uronic acids) were easy to release but unstable, and the release and stability of neutral sugars were in between acidic and basic sugars. In addition, the hydrolysis process was applied to monosaccharide composition analysis of Hippocampus trimaculatus polysaccharide and the appropriate hydrolytic condition was accorded with that of the above four polysaccharides. Thus, different hydrolytic conditions should be used for the monosaccharide composition analysis of polysaccharides based on their structural characteristics. PMID:27083372

  17. Ion temperature by charge exchange neutral analysis from vertical sightlines on the Tokamak Fusion Test Reactor

    SciTech Connect

    Fiore, C.L.; Medley, S.S.; Hammett, G.W.; Kaita, R.; Scott, S.D.

    1987-12-01

    The Fokker-Planck code FPPRF is used to calculate the expected deuterium charge exchange flux along vertical sighlines from TFTR neutral-beam-injected discharges. The feasibility of obtaining central ion temperature measurements by fitting the spectra obtained from these sightlines at two energy regions--above the highest neutral beam injection energy (> 100 keV) and from 20-80 keV--is investigated. It is demonstrated that the central ion temperature can be obtained from the central vertical slightline for fitting the high energy data. The deuterium neutral particle flux energy distribution below the neutral beam injection energy is insentive to the code input ion temperature, however. 6 refs., 12 figs., 1 tab.

  18. Antimicrobial activity of neutralized extracellular culture filtrates of lactic acid bacteria isolated from a cultured Indian milk product ('dahi').

    PubMed

    Varadaraj, M C; Devi, N; Keshava, N; Manjrekar, S P

    1993-12-01

    Neutralized extracellular culture filtrate obtained from isolates of Lactobacillus acidophilus, Lactobacillus delbruecki ssp. bulgaricus, Lactobacillus salivarius and Lactococcus lactis ssp. lactis from 'dahi' showed weak to moderate inhibition of Staphylococcus aureus, Bacillus cereus, Escherichia coli, Bacillus brevis, Bacillus circulans, Bacillus coagulans, Bacillus laterosporus, Bacillus subtilis and Pseudomonas aeruginosa when tested by the diffusion agar well assay method. The effective minimum quantity of lactic culture filtrates required to obtain complete inhibition of an inoculum of 10(3) cfu/ml of the bacteria tested was between 20 and 26% (vol/vol), as determined by the agar incorporation method. Neutralized extracellular culture filtrate of these lactic cultures added at a level of 10% in sterile, 10% reconstituted non-fat dry milk was able to either suppress or retard growth of selected bacterial cultures when incubated at 37 degrees C for 24 h. This study indicated the antimicrobial activity of dahi and the potential of using neutralized extracellular culture filtrate of lactic acid bacteria in the biopreservation of foods.

  19. Canine distemper virus neutralization activity is low in human serum and it is sensitive to an amino acid substitution in the hemagglutinin protein

    SciTech Connect

    Zhang, Xinsheng; Wallace, Olivia L.; Domi, Arban; Wright, Kevin J.; Driscoll, Jonathan; Anzala, Omu; Sanders, Eduard J.; Kamali, Anatoli; Allen, Susan; Fast, Pat; Gilmour, Jill; Price, Matt A.; Parks, Christopher L.

    2015-08-15

    Serum was analyzed from 146 healthy adult volunteers in eastern Africa to evaluate measles virus (MV) and canine distemper virus (CDV) neutralizing antibody (nAb) prevalence and potency. MV plaque reduction neutralization test (PRNT) results indicated that all sera were positive for MV nAbs. Furthermore, the 50% neutralizing dose (ND50) for the majority of sera corresponded to antibody titers induced by MV vaccination. CDV nAbs titers were low and generally were detected in sera with high MV nAb titers. A mutant CDV was generated that was less sensitive to neutralization by human serum. The mutant virus genome had 10 nucleotide substitutions, which coded for single amino acid substitutions in the fusion (F) and hemagglutinin (H) glycoproteins and two substitutions in the large polymerase (L) protein. The H substitution occurred in a conserved region involved in receptor interactions among morbilliviruses, implying that this region is a target for cross-reactive neutralizing antibodies. - Highlights: • Screened 146 serum samples for measles virus (MV) and canine distemper virus (CDV) neutralizing antibody (nAb). • MV nAb is prevalent in the sera. • CDV neutralizing activity is generally low or absent and when detected it is present in sera with high MV nAb titers. • A neutralization-resistant CDV mutant was isolated using human serum selection. • A mutation was identified in the receptor-binding region of CDV hemagglutinin protein that confers the neutralization resistance.

  20. Effects of acute psychosocial stress on neural activity to emotional and neutral faces in a face recognition memory paradigm.

    PubMed

    Li, Shijia; Weerda, Riklef; Milde, Christopher; Wolf, Oliver T; Thiel, Christiane M

    2014-12-01

    Previous studies have shown that acute psychosocial stress impairs recognition of declarative memory and that emotional material is especially sensitive to this effect. Animal studies suggest a central role of the amygdala which modulates memory processes in hippocampus, prefrontal cortex and other brain areas. We used functional magnetic resonance imaging (fMRI) to investigate neural correlates of stress-induced modulation of emotional recognition memory in humans. Twenty-seven healthy, right-handed, non-smoker male volunteers performed an emotional face recognition task. During encoding, participants were presented with 50 fearful and 50 neutral faces. One hour later, they underwent either a stress (Trier Social Stress Test) or a control procedure outside the scanner which was followed immediately by the recognition session inside the scanner, where participants had to discriminate between 100 old and 50 new faces. Stress increased salivary cortisol, blood pressure and pulse, and decreased the mood of participants but did not impact recognition memory. BOLD data during recognition revealed a stress condition by emotion interaction in the left inferior frontal gyrus and right hippocampus which was due to a stress-induced increase of neural activity to fearful and a decrease to neutral faces. Functional connectivity analyses revealed a stress-induced increase in coupling between the right amygdala and the right fusiform gyrus, when processing fearful as compared to neutral faces. Our results provide evidence that acute psychosocial stress affects medial temporal and frontal brain areas differentially for neutral and emotional items, with a stress-induced privileged processing of emotional stimuli.

  1. Application of subtracted gDNA microarray-assisted Bulked Segregant Analysis for rapid discovery of molecular markers associated with day-neutrality in strawberry (Fragaria x ananassa).

    PubMed

    Gor, Mian Chee; Mantri, Nitin; Pang, Edwin

    2016-01-01

    A Fragaria Discovery Panel (FDP; strawberry-specific SDA) containing 287 features was constructed by subtracting the pooled gDNA of nine non-angiosperm species from the pooled gDNA of five strawberry genotypes. This FDP was used for Bulk Segregant Analysis (BSA) to enable identification of molecular markers associated with day-neutrality. Analysis of hybridisation patterns of a short day (SD) DNA bulk and three day-neutral (DN) DNA bulks varying in flowering strength allowed identification of a novel feature, FaP2E11, closely linked to CYTOKININ OXIDASE 1 (CKX1) gene possibly involved in promoting flowering under non-inductive condition. The signal intensities of FaP2E11 feature obtained from the strong DN bulk (DN1) is three fold higher than the short day bulk (SD), indicating that the putative marker may linked to a CKX1 variant allele with lower enzyme activity. We propose a model for flowering regulation based on the hypothesis that flowering strength may be regulated by the copy number of FaP2E11-linked CKX1 alleles. This study demonstrates the feasibility of the SDA-based BSA approach for the identification of molecular markers associated with day-neutrality in strawberry. This innovative strategy is an efficient and cost-effective approach for molecular marker discovery. PMID:27586242

  2. Application of subtracted gDNA microarray-assisted Bulked Segregant Analysis for rapid discovery of molecular markers associated with day-neutrality in strawberry (Fragaria x ananassa).

    PubMed

    Gor, Mian Chee; Mantri, Nitin; Pang, Edwin

    2016-01-01

    A Fragaria Discovery Panel (FDP; strawberry-specific SDA) containing 287 features was constructed by subtracting the pooled gDNA of nine non-angiosperm species from the pooled gDNA of five strawberry genotypes. This FDP was used for Bulk Segregant Analysis (BSA) to enable identification of molecular markers associated with day-neutrality. Analysis of hybridisation patterns of a short day (SD) DNA bulk and three day-neutral (DN) DNA bulks varying in flowering strength allowed identification of a novel feature, FaP2E11, closely linked to CYTOKININ OXIDASE 1 (CKX1) gene possibly involved in promoting flowering under non-inductive condition. The signal intensities of FaP2E11 feature obtained from the strong DN bulk (DN1) is three fold higher than the short day bulk (SD), indicating that the putative marker may linked to a CKX1 variant allele with lower enzyme activity. We propose a model for flowering regulation based on the hypothesis that flowering strength may be regulated by the copy number of FaP2E11-linked CKX1 alleles. This study demonstrates the feasibility of the SDA-based BSA approach for the identification of molecular markers associated with day-neutrality in strawberry. This innovative strategy is an efficient and cost-effective approach for molecular marker discovery.

  3. Application of subtracted gDNA microarray-assisted Bulked Segregant Analysis for rapid discovery of molecular markers associated with day-neutrality in strawberry (Fragaria x ananassa)

    NASA Astrophysics Data System (ADS)

    Gor, Mian Chee; Mantri, Nitin; Pang, Edwin

    2016-09-01

    A Fragaria Discovery Panel (FDP; strawberry-specific SDA) containing 287 features was constructed by subtracting the pooled gDNA of nine non-angiosperm species from the pooled gDNA of five strawberry genotypes. This FDP was used for Bulk Segregant Analysis (BSA) to enable identification of molecular markers associated with day-neutrality. Analysis of hybridisation patterns of a short day (SD) DNA bulk and three day-neutral (DN) DNA bulks varying in flowering strength allowed identification of a novel feature, FaP2E11, closely linked to CYTOKININ OXIDASE 1 (CKX1) gene possibly involved in promoting flowering under non-inductive condition. The signal intensities of FaP2E11 feature obtained from the strong DN bulk (DN1) is three fold higher than the short day bulk (SD), indicating that the putative marker may linked to a CKX1 variant allele with lower enzyme activity. We propose a model for flowering regulation based on the hypothesis that flowering strength may be regulated by the copy number of FaP2E11-linked CKX1 alleles. This study demonstrates the feasibility of the SDA-based BSA approach for the identification of molecular markers associated with day-neutrality in strawberry. This innovative strategy is an efficient and cost-effective approach for molecular marker discovery.

  4. Application of subtracted gDNA microarray-assisted Bulked Segregant Analysis for rapid discovery of molecular markers associated with day-neutrality in strawberry (Fragaria x ananassa)

    PubMed Central

    Gor, Mian Chee; Mantri, Nitin; Pang, Edwin

    2016-01-01

    A Fragaria Discovery Panel (FDP; strawberry-specific SDA) containing 287 features was constructed by subtracting the pooled gDNA of nine non-angiosperm species from the pooled gDNA of five strawberry genotypes. This FDP was used for Bulk Segregant Analysis (BSA) to enable identification of molecular markers associated with day-neutrality. Analysis of hybridisation patterns of a short day (SD) DNA bulk and three day-neutral (DN) DNA bulks varying in flowering strength allowed identification of a novel feature, FaP2E11, closely linked to CYTOKININ OXIDASE 1 (CKX1) gene possibly involved in promoting flowering under non-inductive condition. The signal intensities of FaP2E11 feature obtained from the strong DN bulk (DN1) is three fold higher than the short day bulk (SD), indicating that the putative marker may linked to a CKX1 variant allele with lower enzyme activity. We propose a model for flowering regulation based on the hypothesis that flowering strength may be regulated by the copy number of FaP2E11-linked CKX1 alleles. This study demonstrates the feasibility of the SDA-based BSA approach for the identification of molecular markers associated with day-neutrality in strawberry. This innovative strategy is an efficient and cost-effective approach for molecular marker discovery. PMID:27586242

  5. Fishbone activity in experimental advanced superconducting tokamak neutral beam injection plasma

    SciTech Connect

    Xu, Liqing; Zhang, Jizong; Chen, Kaiyun E-mail: lqhu@ipp.cas.cn; Hu, Liqun E-mail: lqhu@ipp.cas.cn; Li, Erzhong; Lin, Shiyao; Shi, Tonghui; Duan, Yanmin; Zhu, Yubao

    2015-12-15

    Repetitive fishbones near the trapped ion procession frequency were observed for the first time in the neutral beam injection high confinement plasmas in Experimental Advanced Superconducting Tokamak (EAST) tokamak, and diagnosed using a solid-state neutral particle analyzer based on a compact silicon photodiode together with an upgraded high spatial-temporal-resolution multi-arrays soft X-ray (SX) system. This 1/1 typical internal kink mode propagates in the ion-diamagnetism direction with a rotation speed faster than the bulk plasma in the plasma frame. From the SX measurements, this mode frequency is typical of chirping down and the energetic particle effect related to the twisting mode structure. This ion fishbone was found able to trigger a multiple core sawtooth crashes with edge-2/1 sideband modes, as well as to lead to a transition from fishbone to long lived saturated kink mode to fishbone. Furthermore, using SX tomography, a correlation between mode amplitude and mode frequency was found. Finally, a phenomenological prey–predator model was found to reproduce the fishbone nonlinear process well.

  6. Fishbone activity in experimental advanced superconducting tokamak neutral beam injection plasma

    NASA Astrophysics Data System (ADS)

    Xu, Liqing; Zhang, Jizong; Chen, Kaiyun; Hu, Liqun; Li, Erzhong; Lin, Shiyao; Shi, Tonghui; Duan, Yanmin; Zhu, Yubao

    2015-12-01

    Repetitive fishbones near the trapped ion procession frequency were observed for the first time in the neutral beam injection high confinement plasmas in Experimental Advanced Superconducting Tokamak (EAST) tokamak, and diagnosed using a solid-state neutral particle analyzer based on a compact silicon photodiode together with an upgraded high spatial-temporal-resolution multi-arrays soft X-ray (SX) system. This 1/1 typical internal kink mode propagates in the ion-diamagnetism direction with a rotation speed faster than the bulk plasma in the plasma frame. From the SX measurements, this mode frequency is typical of chirping down and the energetic particle effect related to the twisting mode structure. This ion fishbone was found able to trigger a multiple core sawtooth crashes with edge-2/1 sideband modes, as well as to lead to a transition from fishbone to long lived saturated kink mode to fishbone. Furthermore, using SX tomography, a correlation between mode amplitude and mode frequency was found. Finally, a phenomenological prey-predator model was found to reproduce the fishbone nonlinear process well.

  7. Optimizing 50kV hydrogen diagnostic neutral beam performance for active spectroscopy in MST

    NASA Astrophysics Data System (ADS)

    Feng, X.; Boguski, J.; Craig, D.; den Hartog, D. J.; Munaretto, S.; Nornberg, M. D.; Olivia, S.

    2015-11-01

    The 50 kV hydrogen diagnostic neutral beam on MST provides local measurements of impurity ion emission through charge exchange recombination spectroscopy (CHERS) and of core-localized magnetic field through the motional Stark effect (MSE). The beam, which was designed to provide 5A of neutral current at 50 kV to meet these needs, is currently on a test stand to accommodate diagnosis, in order to increase the reliability of beam formation, sustain a steady current of 5 amps for 20ms, and optimize the primary energy fraction. The reliability of arc formation was increased from 40% to 80% success rate with increase of cathode gas pressure from 150kPa to 200kPa, and the stability of the arc current is improved with a decrease of the insulation magnetic field. A calorimeter with 5 thermocouples is installed to measure the horizontal and vertical beam profiles as well as beam divergence. Beam energy components are quantified through Doppler-shift spectroscopy. Preliminary simulation results of the beam using the ALCBEAM code as well as a description of how changes to the beam performance can affect CHERS and MSE measurements are presented. This work is supported by the U.S. DOE.

  8. TRIM21 Immune Signaling Is More Sensitive to Antibody Affinity Than Its Neutralization Activity.

    PubMed

    Foss, Stian; Watkinson, Ruth E; Grevys, Algirdas; McAdam, Martin B; Bern, Malin; Høydahl, Lene Stokken; Dalhus, Bjørn; Michaelsen, Terje E; Sandlie, Inger; James, Leo C; Andersen, Jan Terje

    2016-04-15

    Ab-coated viruses can be detected in the cytosol by the FcR tripartite motif-containing 21 (TRIM21), which rapidly recruits the proteasomal machinery and triggers induction of immune signaling. As such, TRIM21 plays a key role in intracellular protection by targeting invading viruses for destruction and alerting the immune system. A hallmark of immunity is elicitation of a balanced response that is proportionate to the threat, to avoid unnecessary inflammation. In this article, we show how Ab affinity modulates TRIM21 immune function. We constructed a humanized monoclonal IgG1 against human adenovirus type 5 (AdV5) and a panel of Fc-engineered variants with a wide range of affinities for TRIM21. We found that IgG1-coated viral particles were neutralized via TRIM21, even when affinity was reduced by as much as 100-fold. In contrast, induction of NF-κB signaling was more sensitive to reduced affinity between TRIM21 and the Ab variants. Thus, TRIM21 mediates neutralization under suboptimal conditions, whereas induction of immune signaling is balanced according to the functional affinity for the incoming immune stimuli. Our findings have implications for engineering of antiviral IgG therapeutics with tailored effector functions. PMID:26962230

  9. Neutron activation analysis system

    DOEpatents

    Taylor, M.C.; Rhodes, J.R.

    1973-12-25

    A neutron activation analysis system for monitoring a generally fluid media, such as slurries, solutions, and fluidized powders, including two separate conduit loops for circulating fluid samples within the range of radiation sources and detectors is described. Associated with the first loop is a neutron source that emits s high flux of slow and thermal neutrons. The second loop employs a fast neutron source, the flux from which is substantially free of thermal neutrons. Adjacent to both loops are gamma counters for spectrographic determination of the fluid constituents. Other gsmma sources and detectors are arranged across a portion of each loop for deterMining the fluid density. (Official Gazette)

  10. Chord integrated neutral particle diagnostic data analysis for neutral beam injection and ion cyclotron radio frequency heated plasma in a complex Large Helical Device geometry

    SciTech Connect

    Veshchev, E. A.; Goncharov, P. R.; Ozaki, T.; Sudo, S.; Lyon, J. F.

    2006-10-15

    Energy and angle-resolved measurements of charge exchange neutral particle fluxes from the plasma provide information about T{sub i}, as well as non-Maxwellian substantially anisotropic ion distribution tails due to neutral beam injection (NBI) and ion cyclotron radio frequency (ICRF) heating. The measured chord integral neutral flux calculation scheme for the Large Helical Device magnetic surface geometry is given. Calculation results are shown for measurable atomic energy spectra corresponding to heating-induced fast ion distributions from simplified Fokker-Planck models. The behavior of calculated and experimental suprathermal particle distributions from NBI and ICRF heated plasma is discussed in the context of the experimental data interpretation.

  11. Acidic extracellular pH neutralizes the autophagy-inhibiting activity of chloroquine

    PubMed Central

    Pellegrini, Paola; Strambi, Angela; Zipoli, Chiara; Hägg-Olofsson, Maria; Buoncervello, Maria; Linder, Stig; De Milito, Angelo

    2014-01-01

    Acidic pH is an important feature of tumor microenvironment and a major determinant of tumor progression. We reported that cancer cells upregulate autophagy as a survival mechanism to acidic stress. Inhibition of autophagy by administration of chloroquine (CQ) in combination anticancer therapies is currently evaluated in clinical trials. We observed in 3 different human cancer cell lines cultured at acidic pH that autophagic flux is not blocked by CQ. This was consistent with a complete resistance to CQ toxicity in cells cultured in acidic conditions. Conversely, the autophagy-inhibiting activity of Lys-01, a novel CQ derivative, was still detectable at low pH. The lack of CQ activity was likely dependent on a dramatically reduced cellular uptake at acidic pH. Using cell lines stably adapted to chronic acidosis we could confirm that CQ lack of activity was merely caused by acidic pH. Moreover, unlike CQ, Lys-01 was able to kill low pH-adapted cell lines, although higher concentrations were required as compared with cells cultured at normal pH conditions. Notably, buffering medium pH in low pH-adapted cell lines reverted CQ resistance. In vivo analysis of tumors treated with CQ showed that accumulation of strong LC3 signals was observed only in normoxic areas but not in hypoxic/acidic regions. Our observations suggest that targeting autophagy in the tumor environment by CQ may be limited to well-perfused regions but not achieved in acidic regions, predicting possible limitations in efficacy of CQ in antitumor therapies. PMID:24492472

  12. Neutral redox-active hydrogen- and halogen-bonding [2]rotaxanes for the electrochemical sensing of chloride.

    PubMed

    Lim, Jason Y C; Cunningham, Matthew J; Davis, Jason J; Beer, Paul D

    2014-12-14

    The first examples of redox-active ferrocene-functionalised neutral [2]rotaxanes have been synthesised via chloride anion templation. (1)H NMR spectroscopic titrations reveal that these [2]rotaxane host systems recognize chloride selectively over other halides and oxoanions in highly-competitive aqueous media. By replacing the hydrogen bonding prototriazole units of the rotaxane axle component with iodotriazole halogen bond-donor groups, the degree of chloride selectivity of the [2]rotaxanes is modulated. Electrochemical voltammetric experiments demonstrate that the rotaxanes can sense chloride via cathodic perturbations of the respective rotaxanes' ferrocene-ferrocenium redox-couple upon anion addition.

  13. Specifically modified Env immunogens activate B-cell precursors of broadly neutralizing HIV-1 antibodies in transgenic mice

    PubMed Central

    McGuire, Andrew T.; Gray, Matthew D.; Dosenovic, Pia; Gitlin, Alexander D.; Freund, Natalia T.; Petersen, John; Correnti, Colin; Johnsen, William; Kegel, Robert; Stuart, Andrew B.; Glenn, Jolene; Seaman, Michael S.; Schief, William R.; Strong, Roland K.; Nussenzweig, Michel C.; Stamatatos, Leonidas

    2016-01-01

    VRC01-class broadly neutralizing HIV-1 antibodies protect animals from experimental infection and could contribute to an effective vaccine response. Their predicted germline forms (gl) bind Env inefficiently, which may explain why they are not elicited by HIV-1 Env-immunization. Here we show that an optimized Env immunogen can engage multiple glVRC01-class antibodies. Furthermore, this immunogen activates naive B cells expressing the human germline heavy chain of 3BNC60, paired with endogenous mouse light chains in vivo. To address whether it activates B cells expressing the fully humanized gl3BNC60 B-cell receptor (BCR), we immunized mice carrying both the heavy and light chains of gl3BNC60. B cells expressing this BCR display an autoreactive phenotype and fail to respond efficiently to soluble forms of the optimized immunogen, unless it is highly multimerized. Thus, specifically designed Env immunogens can activate naive B cells expressing human BCRs corresponding to precursors of broadly neutralizing HIV-1 antibodies even when the B cells display an autoreactive phenotype. PMID:26907590

  14. In vivo hepatitis B virus-neutralizing activity of an anti-HBsAg humanized antibody in chimpanzees

    PubMed Central

    Kim, Se Ho; Oh, Han Kyu; Ryu, Chun Jeih; Park, Song Yong

    2008-01-01

    Previously, we constructed a humanized antibody (HuS10) that binds to the common a antigenic determinant on the S protein of HBV. In this study, we evaluated its HBV-neutralizing activity in chimpanzees. A study chimpanzee was intravenously administered with a single dose of HuS10, followed by intravenous challenge with the adr subtype of HBV, while a control chimpanzee was only challenged with the virus. The result showed that the control chimpanzee was infected by the virus, and thus serum HBV surface antigen (HBsAg) became positive from the 14th to 20th week and actively acquired serum anti-HBc and anti-HBs antibodies appeared from the 19th and 23rd week, respectively. However, in the case of the study chimpanzee, serum HBsAg became positive from the 34th to 37th week, while actively acquired serum anti-HBc and anti-HBs antibodies appeared from the 37th and 40th week, respectively, indicating that HuS10 neutralized the virus in vivo and thus delayed the HBV infection. This novel humanized antibody will be useful in the immunoprophylaxis of HBV infection. PMID:18305407

  15. Specifically modified Env immunogens activate B-cell precursors of broadly neutralizing HIV-1 antibodies in transgenic mice.

    PubMed

    McGuire, Andrew T; Gray, Matthew D; Dosenovic, Pia; Gitlin, Alexander D; Freund, Natalia T; Petersen, John; Correnti, Colin; Johnsen, William; Kegel, Robert; Stuart, Andrew B; Glenn, Jolene; Seaman, Michael S; Schief, William R; Strong, Roland K; Nussenzweig, Michel C; Stamatatos, Leonidas

    2016-01-01

    VRC01-class broadly neutralizing HIV-1 antibodies protect animals from experimental infection and could contribute to an effective vaccine response. Their predicted germline forms (gl) bind Env inefficiently, which may explain why they are not elicited by HIV-1 Env-immunization. Here we show that an optimized Env immunogen can engage multiple glVRC01-class antibodies. Furthermore, this immunogen activates naive B cells expressing the human germline heavy chain of 3BNC60, paired with endogenous mouse light chains in vivo. To address whether it activates B cells expressing the fully humanized gl3BNC60 B-cell receptor (BCR), we immunized mice carrying both the heavy and light chains of gl3BNC60. B cells expressing this BCR display an autoreactive phenotype and fail to respond efficiently to soluble forms of the optimized immunogen, unless it is highly multimerized. Thus, specifically designed Env immunogens can activate naive B cells expressing human BCRs corresponding to precursors of broadly neutralizing HIV-1 antibodies even when the B cells display an autoreactive phenotype. PMID:26907590

  16. SNP array analysis reveals novel genomic abnormalities including copy neutral loss of heterozygosity in anaplastic oligodendrogliomas.

    PubMed

    Idbaih, Ahmed; Ducray, François; Dehais, Caroline; Courdy, Célia; Carpentier, Catherine; de Bernard, Simon; Uro-Coste, Emmanuelle; Mokhtari, Karima; Jouvet, Anne; Honnorat, Jérôme; Chinot, Olivier; Ramirez, Carole; Beauchesne, Patrick; Benouaich-Amiel, Alexandra; Godard, Joël; Eimer, Sandrine; Parker, Fabrice; Lechapt-Zalcman, Emmanuelle; Colin, Philippe; Loussouarn, Delphine; Faillot, Thierry; Dam-Hieu, Phong; Elouadhani-Hamdi, Selma; Bauchet, Luc; Langlois, Olivier; Le Guerinel, Caroline; Fontaine, Denys; Vauleon, Elodie; Menei, Philippe; Fotso, Marie Janette Motsuo; Desenclos, Christine; Verrelle, Pierre; Verelle, Pierre; Ghiringhelli, François; Noel, Georges; Labrousse, François; Carpentier, Antoine; Dhermain, Frédéric; Delattre, Jean-Yves; Figarella-Branger, Dominique

    2012-01-01

    Anaplastic oligodendrogliomas (AOD) are rare glial tumors in adults with relative homogeneous clinical, radiological and histological features at the time of diagnosis but dramatically various clinical courses. Studies have identified several molecular abnormalities with clinical or biological relevance to AOD (e.g. t(1;19)(q10;p10), IDH1, IDH2, CIC and FUBP1 mutations).To better characterize the clinical and biological behavior of this tumor type, the creation of a national multicentric network, named "Prise en charge des OLigodendrogliomes Anaplasiques (POLA)," has been supported by the Institut National du Cancer (InCA). Newly diagnosed and centrally validated AOD patients and their related biological material (tumor and blood samples) were prospectively included in the POLA clinical database and tissue bank, respectively.At the molecular level, we have conducted a high-resolution single nucleotide polymorphism array analysis, which included 83 patients. Despite a careful central pathological review, AOD have been found to exhibit heterogeneous genomic features. A total of 82% of the tumors exhibited a 1p/19q-co-deletion, while 18% harbor a distinct chromosome pattern. Novel focal abnormalities, including homozygously deleted, amplified and disrupted regions, have been identified. Recurring copy neutral losses of heterozygosity (CNLOH) inducing the modulation of gene expression have also been discovered. CNLOH in the CDKN2A locus was associated with protein silencing in 1/3 of the cases. In addition, FUBP1 homozygous deletion was detected in one case suggesting a putative tumor suppressor role of FUBP1 in AOD.Our study showed that the genomic and pathological analyses of AOD are synergistic in detecting relevant clinical and biological subgroups of AOD. PMID:23071531

  17. SNP Array Analysis Reveals Novel Genomic Abnormalities Including Copy Neutral Loss of Heterozygosity in Anaplastic Oligodendrogliomas

    PubMed Central

    Idbaih, Ahmed; Ducray, François; Dehais, Caroline; Courdy, Célia; Carpentier, Catherine; de Bernard, Simon; Uro-Coste, Emmanuelle; Mokhtari, Karima; Jouvet, Anne; Honnorat, Jérôme; Chinot, Olivier; Ramirez, Carole; Beauchesne, Patrick; Benouaich-Amiel, Alexandra; Godard, Joël; Eimer, Sandrine; Parker, Fabrice; Lechapt-Zalcman, Emmanuelle; Colin, Philippe; Loussouarn, Delphine; Faillot, Thierry; Dam-Hieu, Phong; Elouadhani-Hamdi, Selma; Bauchet, Luc; Langlois, Olivier; Le Guerinel, Caroline; Fontaine, Denys; Vauleon, Elodie; Menei, Philippe; Fotso, Marie Janette Motsuo; Desenclos, Christine; Verelle, Pierre; Ghiringhelli, François; Noel, Georges; Labrousse, François; Carpentier, Antoine; Dhermain, Frédéric; Delattre, Jean-Yves; Figarella-Branger, Dominique

    2012-01-01

    Anaplastic oligodendrogliomas (AOD) are rare glial tumors in adults with relative homogeneous clinical, radiological and histological features at the time of diagnosis but dramatically various clinical courses. Studies have identified several molecular abnormalities with clinical or biological relevance to AOD (e.g. t(1;19)(q10;p10), IDH1, IDH2, CIC and FUBP1 mutations). To better characterize the clinical and biological behavior of this tumor type, the creation of a national multicentric network, named “Prise en charge des OLigodendrogliomes Anaplasiques (POLA),” has been supported by the Institut National du Cancer (InCA). Newly diagnosed and centrally validated AOD patients and their related biological material (tumor and blood samples) were prospectively included in the POLA clinical database and tissue bank, respectively. At the molecular level, we have conducted a high-resolution single nucleotide polymorphism array analysis, which included 83 patients. Despite a careful central pathological review, AOD have been found to exhibit heterogeneous genomic features. A total of 82% of the tumors exhibited a 1p/19q-co-deletion, while 18% harbor a distinct chromosome pattern. Novel focal abnormalities, including homozygously deleted, amplified and disrupted regions, have been identified. Recurring copy neutral losses of heterozygosity (CNLOH) inducing the modulation of gene expression have also been discovered. CNLOH in the CDKN2A locus was associated with protein silencing in 1/3 of the cases. In addition, FUBP1 homozygous deletion was detected in one case suggesting a putative tumor suppressor role of FUBP1 in AOD. Our study showed that the genomic and pathological analyses of AOD are synergistic in detecting relevant clinical and biological subgroups of AOD. PMID:23071531

  18. Reinstatement of pain-related brain activation during the recognition of neutral images previously paired with nociceptive stimuli.

    PubMed

    Forkmann, Katarina; Wiech, Katja; Sommer, Tobias; Bingel, Ulrike

    2015-08-01

    Remembering an event partially reactivates cortical and subcortical brain regions that were engaged during its experience and encoding. Such reinstatement of neuronal activation has been observed in different sensory systems, including the visual, auditory, olfactory, and somatosensory domain. However, so far, this phenomenon of incidental memory has not been explored in the context of pain. In this functional magnetic resonance imaging study, we investigated the neural reinstatement of pain-related and tone-related activations during the recognition of neutral images that had been encoded during (1) painful stimulation, (2) auditory stimulation of comparable unpleasantness, or (3) no additional stimulation. Stimulus-specific reinstatement was tested in 24 healthy male and female participants who performed a visual categorization task (encoding) that was immediately followed by a surprise recognition task. Neural responses were acquired in both sessions. Our data show a partial reinstatement of brain regions frequently associated with pain processing, including the left posterior insula, bilateral putamen, and right operculum, during the presentation of images previously paired with painful heat. This effect was specific to painful stimuli. Moreover, the bilateral ventral striatum showed stronger responses for remembered pain-associated images as compared with tone-associated images, suggesting a higher behavioral relevance of remembering neutral pictures previously paired with pain. Our results support the biological relevance of pain in that only painful but not equally unpleasant auditory stimuli were able to "tag" neutral images during their simultaneous presentation and reactivate pain-related brain regions. Such mechanisms might contribute to the development or maintenance of chronic pain and deserve further investigation in clinical populations.

  19. Glucose administration enhances fMRI brain activation and connectivity related to episodic memory encoding for neutral and emotional stimuli.

    PubMed

    Parent, Marise B; Krebs-Kraft, Desiree L; Ryan, John P; Wilson, Jennifer S; Harenski, Carla; Hamann, Stephan

    2011-04-01

    Glucose enhances memory in a variety of species. In humans, glucose administration enhances episodic memory encoding, although little is known regarding the neural mechanisms underlying these effects. Here we examined whether elevating blood glucose would enhance functional MRI (fMRI) activation and connectivity in brain regions associated with episodic memory encoding and whether these effects would differ depending on the emotional valence of the material. We used a double-blind, within-participants, crossover design in which either glucose (50g) or a saccharin placebo were administered before scanning, on days approximately 1 week apart. We scanned healthy young male participants with fMRI as they viewed emotionally arousing negative pictures and emotionally neutral pictures, intermixed with baseline fixation. Free recall was tested at 5 min after scanning and again after 1 day. Glucose administration increased activation in brain regions associated with successful episodic memory encoding. Glucose also enhanced activation in regions whose activity was correlated with subsequent successful recall, including the hippocampus, prefrontal cortex, and other regions, and these effects differed for negative vs. neutral stimuli. Finally, glucose substantially increased functional connectivity between the hippocampus and amygdala and a network of regions previously implicated in successful episodic memory encoding. These findings fit with evidence from nonhuman animals indicating glucose modulates memory by selectively enhancing neural activity in brain regions engaged during memory tasks. Our results highlight the modulatory effects of glucose and the importance of examining both regional changes in activity and functional connectivity to fully characterize the effects of glucose on brain function and memory.

  20. VEGF neutralizing aerosol therapy in primary pulmonary adenocarcinoma with K-ras activating-mutations.

    PubMed

    Hervé, Virginie; Rabbe, Nathalie; Guilleminault, Laurent; Paul, Flora; Schlick, Laurène; Azzopardi, Nicolas; Duruisseaux, Michael; Fouquenet, Delphine; Montharu, Jérôme; Redini, Françoise; Paintaud, Gilles; Lemarié, Etienne; Cadranel, Jacques; Wislez, Marie; Heuzé-Vourc'h, Nathalie

    2014-01-01

    K-ras mutations promote angiogenesis in lung cancer and contribute to the drug resistance of cancer cells. It is not clear whether K-ras mutated adenocarcinomas are sensitive to anti-angiogenic therapy with monoclonal antibodies (mAbs) that target vascular endothelial growth factor (VEGF). Anti-angiogenic mAbs are usually delivered systemically, but only a small proportion reaches the lung after intravenous injection. We investigated the relevance of a non-invasive pulmonary route for the delivery of anti-VEGF mAbs in the mouse K-ras(LA1) model. We found that pulmonary delivery of these mAbs significantly reduced the number of tumor lesions and inhibited malignant progression. The antitumor effect involves the VEGFR2-dependent inhibition of blood vessel growth, which impairs tumor proliferation. Pharmacokinetic analysis of aerosolized anti-VEGF showed its low rate of passage into the bloodstream, suggesting that this delivery route is associated with reduced systemic side effects. Our findings highlight the value of the aerosol route for administration of anti-angiogenic mAbs in pulmonary adenocarcinoma with K-ras activating-mutations. PMID:25484066

  1. Admissibility analysis for linear singular systems with time-varying delays via neutral system approach.

    PubMed

    Liu, Zhou-Yang; Lin, Chong; Chen, Bing

    2016-03-01

    This paper studies the admissibility problem for a class of linear singular systems with time-varying delays. In order to highlight the relations between the delay and the state, the singular system is transformed into a neutral form. Then, an appropriate type of Lyapunov-Krasovskii functionals is proposed to develop a delay-derivative-dependent admissibility condition in terms of linear matrix inequalities. The derivation combines the Wirtinger-based inequality and reciprocally convex combination method. The present criterion is also for the stability test of retarded and neutral systems with time-varying delays. Some examples are provided to illustrate the effectiveness and the benefits of the proposed method.

  2. Theoretical analysis of some problems in the measurement of beam divergence angle for EAST neutral beam injector

    NASA Astrophysics Data System (ADS)

    Xu, YongJian; Hu, ChunDong

    2011-12-01

    Beam angular divergence is one of the indicators to evaluate the beam quality. Operating parameters of the beam extraction system could be adjusted to gain better beam quality following the measurement results, which will be helpful not only to study the transmission characteristics of the beam and the power distribution on the heat load components, but also to understand the real-time working condition of the ion source and beam extraction system. This study includes: (1) the theoretical analysis of beam extraction pulse duration for measurement of beam angular divergence; (2) the theoretical analysis of beam intensity distribution during beam transmission for Experimental Advanced Superconducting Tokomak (EAST) neutral beam injector. Those theoretical analyses could point the way to the measurement of beam divergence angle for EAST neutral beam injector.

  3. Divergent Trends of Anti-JCPyV Serum Reactivity and Neutralizing Activity in Multiple Sclerosis (MS) Patients during Treatment with Natalizumab

    PubMed Central

    Diotti, Roberta Antonia; Capra, Ruggero; Moiola, Lucia; Caputo, Valeria; De Rossi, Nicola; Sangalli, Francesca; Martinelli, Vittorio; Burioni, Roberto; Clementi, Massimo; Mancini, Nicasio

    2016-01-01

    The association between natalizumab and progressive multifocal leukoencephalopathy (PML) is established, but a reliable clinical risk stratification flow-chart is lacking. New risk factors are needed, such as the possible role of the anti-JC polyomavirus (JCPyV) neutralizing antibody. In this pilot study, we analyzed this parameter during natalizumab treatment. Sequential sera of 38 multiple sclerosis patients during their first year of natalizumab treatment were collected, and grouped according to the number of infusions. For 11 patients, samples were also available after 24 infusions (T24), when progressive multifocal leukoencephalopathy (PML) risk is higher. The reactivity against VP1, the main JCPyV surface protein, and the anti-JCPyV neutralizing activity were evaluated. During the first year, a lack of correlation between anti-JCPyV antibody response and its neutralizing activity was observed: a significant decrease in anti-JCPyV antibody response was observed (p = 0.0039), not paralleled by a similar trend in the total anti-JCPyV neutralizing activity (p = 0.2239). This lack of correlation was even more evident at T24 when, notwithstanding a significant increase in the anti-JCPyV response (p = 0.0097), a further decrease of the neutralizing activity was observed (p = 0.0062). This is the first study evidencing, prospectively, the lack of correlation between the anti-JCPyV antibody response and its neutralizing activity during natalizumab treatment. PMID:27164128

  4. Divergent Trends of Anti-JCPyV Serum Reactivity and Neutralizing Activity in Multiple Sclerosis (MS) Patients during Treatment with Natalizumab.

    PubMed

    Diotti, Roberta Antonia; Capra, Ruggero; Moiola, Lucia; Caputo, Valeria; De Rossi, Nicola; Sangalli, Francesca; Martinelli, Vittorio; Burioni, Roberto; Clementi, Massimo; Mancini, Nicasio

    2016-01-01

    The association between natalizumab and progressive multifocal leukoencephalopathy (PML) is established, but a reliable clinical risk stratification flow-chart is lacking. New risk factors are needed, such as the possible role of the anti-JC polyomavirus (JCPyV) neutralizing antibody. In this pilot study, we analyzed this parameter during natalizumab treatment. Sequential sera of 38 multiple sclerosis patients during their first year of natalizumab treatment were collected, and grouped according to the number of infusions. For 11 patients, samples were also available after 24 infusions (T24), when progressive multifocal leukoencephalopathy (PML) risk is higher. The reactivity against VP1, the main JCPyV surface protein, and the anti-JCPyV neutralizing activity were evaluated. During the first year, a lack of correlation between anti-JCPyV antibody response and its neutralizing activity was observed: a significant decrease in anti-JCPyV antibody response was observed (p = 0.0039), not paralleled by a similar trend in the total anti-JCPyV neutralizing activity (p = 0.2239). This lack of correlation was even more evident at T24 when, notwithstanding a significant increase in the anti-JCPyV response (p = 0.0097), a further decrease of the neutralizing activity was observed (p = 0.0062). This is the first study evidencing, prospectively, the lack of correlation between the anti-JCPyV antibody response and its neutralizing activity during natalizumab treatment. PMID:27164128

  5. Effect of delayed anthrax vaccine dose on Bacillus anthracis protective antigen IgG response and lethal toxin neutralization activity.

    PubMed

    Pittman, Phillip R; Fisher, Diana; Quinn, Xiaofei; Schmader, Trevor; Barrera-Oro, Julio G

    2013-10-17

    We describe the Bacillus anthracis protective antigen IgG antibody response and the B. anthracis lethal toxin neutralization activity to a delayed dose of anthrax vaccine adsorbed (AVA, BioThrax(®)) using validated assays. 373 individuals received 1, 2, or 3 priming doses, 18-24 months afterward, they received a delayed dose of AVA. Overall, 23.6% of subjects showed detectable anti-PA IgG before the boost, compared to 99.2% (P<0.0001) 28 days after the boost. Geometric mean anti-PA IgG concentration (GMC) was 1.66 μg/mL before and 887.82 μg/mL after the boost (P<0.0001). The proportion of individuals with four-fold increase in GMC following the boost ranged from 93.8% to 100%. Robust anti-PA IgG levels and B. anthracis lethal toxin neutralization activity are induced when an AVA dose is delayed as long as two years. These data support continuing with the vaccination schedule when a dose is delayed as long as two years rather than restarting the series.

  6. Orientation Control in Thin Films of a High-χ Block Copolymer with a Surface Active Embedded Neutral Layer.

    PubMed

    Zhang, Jieqian; Clark, Michael B; Wu, Chunyi; Li, Mingqi; Trefonas, Peter; Hustad, Phillip D

    2016-01-13

    Directed self-assembly (DSA) of block copolymers (BCPs) is an attractive advanced patterning technology being considered for future integrated circuit manufacturing. By controlling interfacial interactions, self-assembled microdomains in thin films of polystyrene-block-poly(methyl methacrylate), PS-b-PMMA, can be oriented perpendicular to surfaces to form line/space or hole patterns. However, its relatively weak Flory interaction parameter, χ, limits its capability to pattern sub-10 nm features. Many BCPs with higher interaction parameters are capable of forming smaller features, but these "high-χ" BCPs typically have an imbalance in surface energy between the respective blocks that make it difficult to achieve the required perpendicular orientation. To address this challenge, we devised a polymeric surface active additive mixed into the BCP solution, referred to as an embedded neutral layer (ENL), which segregates to the top of the BCP film during casting and annealing and balances the surface tensions at the top of the thin film. The additive comprises a second BCP with a "neutral block" designed to provide matched surface tensions with the respective polymers of the main BCP and a "surface anchoring block" with very low surface energy that drives the material to the air interface during spin-casting and annealing. The surface anchoring block allows the film to be annealed above the glass transition temperature of the two materials without intermixing of the two components. DSA was also demonstrated with this embedded neutral top layer formulation on a chemical patterned template using a single step coat and simple thermal annealing. This ENL technology holds promise to enable the use of high-χ BCPs in advanced patterning applications.

  7. Orientation Control in Thin Films of a High-χ Block Copolymer with a Surface Active Embedded Neutral Layer.

    PubMed

    Zhang, Jieqian; Clark, Michael B; Wu, Chunyi; Li, Mingqi; Trefonas, Peter; Hustad, Phillip D

    2016-01-13

    Directed self-assembly (DSA) of block copolymers (BCPs) is an attractive advanced patterning technology being considered for future integrated circuit manufacturing. By controlling interfacial interactions, self-assembled microdomains in thin films of polystyrene-block-poly(methyl methacrylate), PS-b-PMMA, can be oriented perpendicular to surfaces to form line/space or hole patterns. However, its relatively weak Flory interaction parameter, χ, limits its capability to pattern sub-10 nm features. Many BCPs with higher interaction parameters are capable of forming smaller features, but these "high-χ" BCPs typically have an imbalance in surface energy between the respective blocks that make it difficult to achieve the required perpendicular orientation. To address this challenge, we devised a polymeric surface active additive mixed into the BCP solution, referred to as an embedded neutral layer (ENL), which segregates to the top of the BCP film during casting and annealing and balances the surface tensions at the top of the thin film. The additive comprises a second BCP with a "neutral block" designed to provide matched surface tensions with the respective polymers of the main BCP and a "surface anchoring block" with very low surface energy that drives the material to the air interface during spin-casting and annealing. The surface anchoring block allows the film to be annealed above the glass transition temperature of the two materials without intermixing of the two components. DSA was also demonstrated with this embedded neutral top layer formulation on a chemical patterned template using a single step coat and simple thermal annealing. This ENL technology holds promise to enable the use of high-χ BCPs in advanced patterning applications. PMID:26682931

  8. Improved volatile fatty acids anaerobic production from waste activated sludge by pH regulation: Alkaline or neutral pH?

    PubMed

    Ma, Huijun; Chen, Xingchun; Liu, He; Liu, Hongbo; Fu, Bo

    2016-02-01

    In this study, the anaerobic fermentation was carried out for volatile fatty acids (VFAs) production at different pH (between 7.0 and 10.0) conditions with untreated sludge and heat-alkaline pretreated waste activated sludge. In the fermentation with untreated sludge, the extent of hydrolysis of organic matters and extent of acidification at alkaline pH are 54.37% and 30.37%, respectively, resulting in the highest VFAs yield at 235.46mg COD/gVS of three pH conditions. In the fermentation with heat-alkaline pretreated sludge, the acidification rate and VFAs yield at neutral pH are 30.98% and 240.14mg COD/gVS, respectively, which are higher than that at other pH conditions. With the glucose or bovine serum albumin as substrate for VFAs production, the neutral pH showed a higher VFAs concentration than the alkaline pH condition. The results of terminal restriction fragment length polymorphism (T-RFLP) analysis indicated that the alkaline pH caused low microbial richness. Based on the results in this study, we demonstrated that the alkaline pH is favor of hydrolysis of organic matter in sludge while neutral pH improved the acidogenesis for the VFAs production from sludge. Our finding is obvious different to the previous research and helpful for the understanding of how heat-alkaline pretreatment and alkaline fermentation influence the VFAs production, and beneficial to the development of VFAs production process.

  9. Cross-Neutralization Activity of Single-Chain Variable Fragment (scFv) Derived from Anti-V3 Monoclonal Antibodies Mediated by Post-Attachment Binding.

    PubMed

    Maruta, Yasuhiro; Kuwata, Takeo; Tanaka, Kazuki; Alam, Muntasir; Valdez, Kristel Paola Ramirez; Egami, Yoshika; Suwa, Yoshiaki; Morioka, Hiroshi; Matsushita, Shuzo

    2016-09-21

    The V3 loop in the envelope (Env) of HIV-1 is one of the major targets of neutralizing antibodies. However, this antigen is hidden inside the Env trimer in most isolates and is fully exposed only during CD4-gp120 interaction. Thus, primary HIV-1 isolates are relatively resistant to anti-V3 antibodies because IgG is too large to access the V3 loop. To overcome this obstacle, we constructed single-chain variable fragments (scFvs) from anti-V3 monoclonal antibodies 0.5γ, 5G2, and 16G6. Enhanced neutralization by 0.5γ and 5G2 scFvs was observed in strains resistant to their IgG counterparts. Neutralization coverage by 0.5γ scFv reached up to 90% of the tested viruses (tier 2 and 3 classes). The temperature-regulated neutralization assay revealed that extensive cross-neutralization of 0.5γ scFv can be explained by post-attachment neutralization. Neutralization assay involving viruses carrying an inter-subunit disulfide bond (SOS virus) showed that the neutralization-susceptible timeframe after attachment was 60 to 120 min. These results indicate that the scFvs efficiently access the V3 loop and subsequently neutralize HIV-1, even after virus attachment to the target cells. Based on its broad and potent neutralizing activity, further development of anti-V3 scFv for therapeutic and preventive strategies is warranted.

  10. A fully human monoclonal antibody with novel binding epitope and excellent neutralizing activity to multiple human IFN-α subtypes: A candidate therapy for systemic lupus erythematosus.

    PubMed

    Du, Peng; Xu, Lei; Qiu, Weiyi; Zeng, Dadi; Yue, Junjie; Wang, Shuang; Huang, Peitang; Sun, Zhiwei

    2015-01-01

    Systemic lupus erythematosus (SLE) is a chronic, heterogeneous autoimmune disease short of effective therapeutic agents. A multitude of studies of SLE in the last decade have accentuated a central role of the interferon alpha (IFN-α) pathway in SLE pathogenesis. We report here a candidate therapeutic neutralizing antibody, AIA22, with a different binding epitope and discrepant neutralizing profile from the anti-multiple IFN-α subtype antibodies currently in clinical trials. AIA22 specifically interacts with multiple IFN-α subtypes, binds to the type I IFN receptor 2 (IFNAR2) recognition region of IFN-α (considered a novel antigen epitope), and effectively neutralizes the activity of almost all of the IFN-α subtypes (with the exception of IFN-α7) both in vitro and in vivo. Concurrently, structural modeling and computational design yielded a mutational antibody of AIA22, AIAmut, which exhibited substantially improved neutralizing activity to multiple IFN-α subtypes.

  11. Methane oxidation activity and diversity of aerobic methanotrophs in pH-neutral and semi-neutral thermal springs of the Kunashir Island, Russian Far East.

    PubMed

    Kizilova, A K; Sukhacheva, M V; Pimenov, N V; Yurkov, A M; Kravchenko, I K

    2014-03-01

    Aerobic methane oxidation has been mostly studied in environments with moderate to low temperatures. However, the process also occurs in terrestrial thermal springs, where little research on the subject has been done to date. The potential activity of methane oxidation and diversity of aerobic methanotrophic bacteria were studied in sediments of thermal springs with various chemical and physical properties, sampled across the Kunashir Island, the Kuriles archipelago. Activity was measured by means of the radioisotope tracer technique utilizing (14)C-labeled methane. Biodiversity assessments were based on the particulate methane monooxygenase (pmoA) gene, which is found in all known thermophilic and thermotolerant methanotrophs. We demonstrated the possibility of methane oxidation in springs with temperature exceeding 74 °C, and the most intensive methane uptake was shown in springs with temperatures about 46 °C. PmoA was detected in 19 out of 30 springs investigated and the number of pmoA gene copies varied between 10(4) and 10(6) copies per ml of sediment. Phylogenetic analysis of PmoA sequences revealed the presence of methanotrophs from both the Alpha- and Gammaproteobacteria. Our results suggest that methanotrophs inhabiting thermal springs with temperature exceeding 50 °C may represent novel thermophilic and thermotolerant species of the genera Methylocystis and Methylothermus, as well as previously undescribed Gammaproteobacteria. PMID:24343375

  12. Neutralization of plasminogen activator inhibitor I (PAI-1) by the synthetic antagonist PAI-749 via a dual mechanism of action.

    PubMed

    Gardell, Stephen J; Krueger, Julie A; Antrilli, Thomas A; Elokdah, Hassan; Mayer, Scott; Orcutt, Steven J; Crandall, David L; Vlasuk, George P

    2007-10-01

    PAI-749 is a potent and selective synthetic antagonist of plasminogen activator inhibitor 1 (PAI-1) that preserved tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA) activities in the presence of PAI-1 (IC(50) values, 157 and 87 nM, respectively). The fluorescence (Fl) of fluorophore-tagged PAI-1 (PAI-NBD119) was quenched by PAI-749; the apparent K(d) (254 nM) was similar to the IC(50) (140 nM) for PAI-NBD119 inactivation. PAI-749 analogs displayed the same potency rank order for neutralizing PAI-1 activity and perturbing PAI-NBD119 Fl; hence, binding of PAI-749 to PAI-1 and inactivation of PAI-1 activity are tightly linked. Exposure of PAI-1 to PAI-749 for 5 min (sufficient for full inactivation) followed by PAI-749 sequestration with Tween 80 micelles yielded active PAI-1; thus, PAI-749 did not irreversibly inactivate PAI-1, a known metastable protein. Treatment of PAI-1 with a PAI-749 homolog (producing less assay interference) blocked the ability of PAI-1 to displace p-aminobenzamidine from the uPA active site. Consistent with this observation, PAI-749 abolished formation of the SDS-stable tPA/PAI-1 complex. PAI-749-mediated neutralization of PAI-1 was associated with induction of PAI-1 polymerization as assessed by native gel electrophoresis. PAI-749 did not turn PAI-1 into a substrate for tPA; however, PAI-749 promoted plasmin-mediated degradation of PAI-1. In conclusion, PAI-1 inactivation by PAI-749 using purified components can result from a dual mechanism of action. First, PAI-749 binds directly to PAI-1, blocks PAI-1 from accessing the active site of tPA, and abrogates formation of the SDS-stable tPA/PAI-1 complex. Second, binding of PAI-749 to PAI-1 renders PAI-1 vulnerable to plasmin-mediated proteolytic degradation. PMID:17622579

  13. Cryo-EM structures elucidate neutralizing mechanisms of anti-chikungunya human monoclonal antibodies with therapeutic activity

    PubMed Central

    Long, Feng; Fong, Rachel H.; Austin, Stephen K.; Chen, Zhenguo; Klose, Thomas; Fokine, Andrei; Liu, Yue; Porta, Jason; Sapparapu, Gopal; Akahata, Wataru; Doranz, Benjamin J.; Crowe, James E.; Diamond, Michael S.; Rossmann, Michael G.

    2015-01-01

    Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that causes severe acute and chronic disease in humans. Although highly inhibitory murine and human monoclonal antibodies (mAbs) have been generated, the structural basis of their neutralizing activity remains poorly characterized. Here, we determined the cryo-EM structures of chikungunya virus-like particles complexed with antibody fragments (Fab) of two highly protective human mAbs, 4J21 and 5M16, that block virus fusion with host membranes. Both mAbs bind primarily to sites within the A and B domains, as well as to the B domain’s β-ribbon connector of the viral glycoprotein E2. The footprints of these antibodies on the viral surface were consistent with results from loss-of-binding studies using an alanine scanning mutagenesis-based epitope mapping approach. The Fab fragments stabilized the position of the B domain relative to the virus, particularly for the complex with 5M16. This finding is consistent with a mechanism of neutralization in which anti-CHIKV mAbs that bridge the A and B domains impede movement of the B domain away from the underlying fusion loop on the E1 glycoprotein and therefore block the requisite pH-dependent fusion of viral and host membranes. PMID:26504196

  14. Chlamydial Plasmid-Encoded Virulence Factor Pgp3 Neutralizes the Antichlamydial Activity of Human Cathelicidin LL-37

    PubMed Central

    Hou, Shuping; Dong, Xiaohua; Yang, Zhangsheng; Li, Zhongyu; Liu, Quanzhong

    2015-01-01

    Chlamydia trachomatis infection in the lower genital tract can ascend to and cause pathologies in the upper genital tract, potentially leading to severe complications, such as tubal infertility. However, chlamydial organisms depleted of plasmid or deficient in the plasmid-encoded Pgp3 are attenuated in ascending infection and no longer are able to induce the upper genital tract pathologies, indicating a significant role of Pgp3 in chlamydial pathogenesis. We now report that C. trachomatis Pgp3 can neutralize the antichlamydial activity of human cathelicidin LL-37, a host antimicrobial peptide secreted by both genital tract epithelial cells and infiltrating neutrophils. Pgp3 bound to and formed stable complexes with LL-37. We further showed that the middle region of Pgp3 (Pgp3m) was responsible for both the binding to and neutralization of LL-37, suggesting that Pgp3m can be targeted for attenuating chlamydial pathogenicity or developed for blocking LL-37-involved non-genital-tract pathologies, such as rosacea and psoriasis. Thus, the current study has provided significant information for both understanding the mechanisms of chlamydial pathogenesis and developing novel therapeutic agents. PMID:26416907

  15. Cryo-EM structures elucidate neutralizing mechanisms of anti-chikungunya human monoclonal antibodies with therapeutic activity.

    PubMed

    Long, Feng; Fong, Rachel H; Austin, Stephen K; Chen, Zhenguo; Klose, Thomas; Fokine, Andrei; Liu, Yue; Porta, Jason; Sapparapu, Gopal; Akahata, Wataru; Doranz, Benjamin J; Crowe, James E; Diamond, Michael S; Rossmann, Michael G

    2015-11-10

    Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that causes severe acute and chronic disease in humans. Although highly inhibitory murine and human monoclonal antibodies (mAbs) have been generated, the structural basis of their neutralizing activity remains poorly characterized. Here, we determined the cryo-EM structures of chikungunya virus-like particles complexed with antibody fragments (Fab) of two highly protective human mAbs, 4J21 and 5M16, that block virus fusion with host membranes. Both mAbs bind primarily to sites within the A and B domains, as well as to the B domain's β-ribbon connector of the viral glycoprotein E2. The footprints of these antibodies on the viral surface were consistent with results from loss-of-binding studies using an alanine scanning mutagenesis-based epitope mapping approach. The Fab fragments stabilized the position of the B domain relative to the virus, particularly for the complex with 5M16. This finding is consistent with a mechanism of neutralization in which anti-CHIKV mAbs that bridge the A and B domains impede movement of the B domain away from the underlying fusion loop on the E1 glycoprotein and therefore block the requisite pH-dependent fusion of viral and host membranes. PMID:26504196

  16. Neutral sphingomyelinase-2 is a redox sensitive enzyme: role of catalytic cysteine residues in regulation of enzymatic activity through changes in oligomeric state.

    PubMed

    Dotson, P Patrick; Karakashian, Alexander A; Nikolova-Karakashian, Mariana N

    2015-02-01

    Neutral sphingomyelinase-2 (nSMase-2) is the major sphingomyelinase activated in response to pro-inflammatory cytokines and during oxidative stress. It is a membrane-bound 655 amino acid protein containing 22 cysteine residues. In this study, we expressed recombinant mouse nSMase-2 protein in Escherichia coli, and investigated whether nSMase-2 is a redox sensitive enzyme. Our results demonstrate that nSMase-2 exists as both monomers and multimers that are associated with high and low enzymatic activity respectively. Mutational analysis of nSMase-2 identified within its C-terminal catalytic domain several oxidant-sensitive cysteine residues that were shown to be involved in enzyme oligomerization. Changing Cys(617) to Ser for example is a gain-of-function mutation associated with a decreased propensity for oligomerization. Alternatively, nSMase-2 expression in a bacterial strain that lacks endogenous thioredoxin, Rosetta-gami2, results in increased oligomer formation and lower enzyme activity. Phenotypic rescue was accomplished by treating nSMase-2 lysates with recombinant human thioredoxin. This indicates that nSMase-2 may be a novel substrate for thioredoxin. FRET analysis confirmed the presence of nSMase-2 multimers in mammalian HEK cells and their localization to the plasma membrane. In conclusion, our results identify nSMase-2 as a redox-sensitive enzyme, whose basal activity is influenced by thioredoxin-mediated changes in its oligomeric state. PMID:25287744

  17. A mouse monoclonal antibody against dengue virus type 1 Mochizuki strain targeting envelope protein domain II and displaying strongly neutralizing but not enhancing activity.

    PubMed

    Yamanaka, Atsushi; Kotaki, Tomohiro; Konishi, Eiji

    2013-12-01

    Dengue fever and its more severe form, dengue hemorrhagic fever, are major global concerns. Infection-enhancing antibodies are major factors hypothetically contributing to increased disease severity. In this study, we generated 26 monoclonal antibodies (MAbs) against the dengue virus type 1 Mochizuki strain. We selected this strain because a relatively large number of unique and rare amino acids were found on its envelope protein. Although most MAbs showing neutralizing activities exhibited enhancing activities at subneutralizing doses, one MAb (D1-IV-7F4 [7F4]) displayed neutralizing activities without showing enhancing activities at lower concentrations. In contrast, another MAb (D1-V-3H12 [3H12]) exhibited only enhancing activities, which were suppressed by pretreatment of cells with anti-FcγRIIa. Although antibody engineering revealed that antibody subclass significantly affected 7F4 (IgG3) and 3H12 (IgG1) activities, neutralizing/enhancing activities were also dependent on the epitope targeted by the antibody. 7F4 recognized an epitope on the envelope protein containing E118 (domain II) and had a neutralizing activity 10- to 1,000-fold stronger than the neutralizing activity of previously reported human or humanized neutralizing MAbs targeting domain I and/or domain II. An epitope-blocking enzyme-linked immunosorbent assay (ELISA) indicated that a dengue virus-immune population possessed antibodies sharing an epitope with 7F4. Our results demonstrating induction of these antibody species (7F4 and 3H12) in Mochizuki-immunized mice may have implications for dengue vaccine strategies designed to minimize induction of enhancing antibodies in vaccinated humans. PMID:24049185

  18. A novel neutral, halophile Stachybotrys microspora-based endoglucanase active impact on β-glucan.

    PubMed

    Benhmad, Ines; Boudabbous, Manel; Yaîch, Asma; Rebai, Maryem; Gargouri, Ali

    2016-04-01

    The production of cellulases from Stachybotrys microspora strain (A19) has been improved by fed-batch fermentation on Avicel cellulose 10 mg/ml. An endoglucanase EG2 was purified to homogeneity. This cellulase has a molecular mass estimated to 50 kDa when analyzed by a denaturant gel electrophoresis. It exhibited an optimal activity at 50 °C, pH 7.0 and 0.85 M NaCl. Specifically, these results show the thermo-active, alkali-tolerant and halo-tolerant properties of EG2. In addition, this endoglucanase showed its highest activity on barley-β-glucan, compared to the CMC. Moreover, it was less active on Avicel cellulose. Furthermore, the EG2 activity was stimulated in the presence of EDTA, urea and β-mercaptoethanol whereas it was reduced in the presence of SDS. This cellulase was highly stable in the presence of organic solvents such as acetone and n-hexane. TLC showed that the main hydrolysis products from EG2 were cellobiose and glucose. This fungal endoglucanase could be potentially important in the conversion of grass-derived biomass into fermentable sugars. PMID:26861652

  19. A novel neutral, halophile Stachybotrys microspora-based endoglucanase active impact on β-glucan.

    PubMed

    Benhmad, Ines; Boudabbous, Manel; Yaîch, Asma; Rebai, Maryem; Gargouri, Ali

    2016-04-01

    The production of cellulases from Stachybotrys microspora strain (A19) has been improved by fed-batch fermentation on Avicel cellulose 10 mg/ml. An endoglucanase EG2 was purified to homogeneity. This cellulase has a molecular mass estimated to 50 kDa when analyzed by a denaturant gel electrophoresis. It exhibited an optimal activity at 50 °C, pH 7.0 and 0.85 M NaCl. Specifically, these results show the thermo-active, alkali-tolerant and halo-tolerant properties of EG2. In addition, this endoglucanase showed its highest activity on barley-β-glucan, compared to the CMC. Moreover, it was less active on Avicel cellulose. Furthermore, the EG2 activity was stimulated in the presence of EDTA, urea and β-mercaptoethanol whereas it was reduced in the presence of SDS. This cellulase was highly stable in the presence of organic solvents such as acetone and n-hexane. TLC showed that the main hydrolysis products from EG2 were cellobiose and glucose. This fungal endoglucanase could be potentially important in the conversion of grass-derived biomass into fermentable sugars.

  20. Identification, cloning, and characterization of a novel soluble receptor that binds IL-22 and neutralizes its activity.

    PubMed

    Kotenko, S V; Izotova, L S; Mirochnitchenko, O V; Esterova, E; Dickensheets, H; Donnelly, R P; Pestka, S

    2001-06-15

    With the use of a partial sequence of the human genome, we identified a gene encoding a novel soluble receptor belonging to the class II cytokine receptor family. This gene is positioned on chromosome 6 in the vicinity of the IFNGR1 gene in a head-to-tail orientation. The gene consists of six exons and encodes a 231-aa protein with a 21-aa leader sequence. The secreted mature protein demonstrates 34% amino acid identity to the extracellular domain of the IL-22R1 chain. Cross-linking experiments demonstrate that the protein binds IL-22 and prevents binding of IL-22 to the functional cell surface IL-22R complex, which consists of two subunits, the IL-22R1 and the IL-10R2c chains. Moreover, this soluble receptor, designated IL-22-binding protein (BP), is capable of neutralizing IL-22 activity. In the presence of the IL-22BP, IL-22 is unable to induce Stat activation in IL-22-responsive human lung carcinoma A549 cells. IL-22BP also blocked induction of the suppressors of cytokine signaling-3 (SOCS-3) gene expression by IL-22 in HepG2 cells. To further evaluate IL-22BP action, we used hamster cells expressing a modified IL-22R complex consisting of the intact IL-10R2c and the chimeric IL-22R1/gammaR1 receptor in which the IL-22R1 intracellular domain was replaced with the IFN-gammaR1 intracellular domain. In these cells, IL-22 activates biological activities specific for IFN-gamma, such as up-regulation of MHC class I Ag expression. The addition of IL-22BP neutralizes the ability of IL-22 to induce Stat activation and MHC class I Ag expression in these cells. Thus, the soluble receptor designated IL-22BP inhibits IL-22 activity by binding IL-22 and blocking its interaction with the cell surface IL-22R complex.

  1. CTEPP STANDARD OPERATING PROCEDURE FOR EXTRACTING AND PREPARING DUST AND SOIL SAMPLES FOR ANALYSIS OF NEUTRAL PERSISTENT ORGANIC POLLUTANTS (SOP-5.14)

    EPA Science Inventory

    This SOP summarizes the method for extracting and preparing a dust or soil sample for analysis of neutral persistent organic pollutants. It covers the extraction and concentration of samples that are to be analyzed by gas chromatography/mass spectrometry.

  2. Endotoxin-neutralizing activity and mechanism of action of a cationic α-helical antimicrobial octadecapeptide derived from α-amylase of rice.

    PubMed

    Taniguchi, Masayuki; Ochiai, Akihito; Matsushima, Kenta; Tajima, Koji; Kato, Tetsuo; Saitoh, Eiichi; Tanaka, Takaaki

    2016-01-01

    We have previously reported that AmyI-1-18, an octadecapeptide derived from α-amylase (AmyI-1) of rice, is a novel cationic α-helical peptide that exhibited antimicrobial activity against human pathogens, including Porphyromonas gingivalis, Pseudomonas aeruginosa, Propionibacterium acnes, Streptococcus mutans, and Candida albicans. In this study, to further investigate the potential functions of AmyI-1-18, we examined its inhibitory ability against the endotoxic activities of lipopolysaccharides (LPSs, smooth and Rc types) and lipid A from Escherichia coli. AmyI-1-18 inhibited the production of endotoxin-induced nitric oxide (NO), an inflammatory mediator, in mouse macrophages (RAW264) in a concentration-dependent manner. The results of a chromogenic Limulus amebocyte lysate assay illustrated that the ability [50% effective concentration (EC50): 0.17 μM] of AmyI-1-18 to neutralize lipid A was similar to its ability (EC50: 0.26 μM) to neutralize LPS, suggesting that AmyI-1-18 specifically binds to the lipid A moiety of LPS. Surface plasmon resonance analysis of the interaction between AmyI-1-18 and LPS or lipid A also suggested that AmyI-1-18 directly binds to the lipid A moiety of LPS because the dissociation constant (KD) of AmyI-1-18 with lipid A is 5.6×10(-10) M, which is similar to that (4.3×10(-10) M) of AmyI-1-18 with LPS. In addition, AmyI-1-18 could block the binding of LPS-binding protein to LPS, although its ability was less than that of polymyxin B. These results suggest that AmyI-1-18 expressing antimicrobial and endotoxin-neutralizing activities is useful as a safe and potent host defense peptide against pathogenic Gram-negative bacteria in many fields of healthcare. PMID:26643956

  3. Endotoxin-neutralizing activity and mechanism of action of a cationic α-helical antimicrobial octadecapeptide derived from α-amylase of rice.

    PubMed

    Taniguchi, Masayuki; Ochiai, Akihito; Matsushima, Kenta; Tajima, Koji; Kato, Tetsuo; Saitoh, Eiichi; Tanaka, Takaaki

    2016-01-01

    We have previously reported that AmyI-1-18, an octadecapeptide derived from α-amylase (AmyI-1) of rice, is a novel cationic α-helical peptide that exhibited antimicrobial activity against human pathogens, including Porphyromonas gingivalis, Pseudomonas aeruginosa, Propionibacterium acnes, Streptococcus mutans, and Candida albicans. In this study, to further investigate the potential functions of AmyI-1-18, we examined its inhibitory ability against the endotoxic activities of lipopolysaccharides (LPSs, smooth and Rc types) and lipid A from Escherichia coli. AmyI-1-18 inhibited the production of endotoxin-induced nitric oxide (NO), an inflammatory mediator, in mouse macrophages (RAW264) in a concentration-dependent manner. The results of a chromogenic Limulus amebocyte lysate assay illustrated that the ability [50% effective concentration (EC50): 0.17 μM] of AmyI-1-18 to neutralize lipid A was similar to its ability (EC50: 0.26 μM) to neutralize LPS, suggesting that AmyI-1-18 specifically binds to the lipid A moiety of LPS. Surface plasmon resonance analysis of the interaction between AmyI-1-18 and LPS or lipid A also suggested that AmyI-1-18 directly binds to the lipid A moiety of LPS because the dissociation constant (KD) of AmyI-1-18 with lipid A is 5.6×10(-10) M, which is similar to that (4.3×10(-10) M) of AmyI-1-18 with LPS. In addition, AmyI-1-18 could block the binding of LPS-binding protein to LPS, although its ability was less than that of polymyxin B. These results suggest that AmyI-1-18 expressing antimicrobial and endotoxin-neutralizing activities is useful as a safe and potent host defense peptide against pathogenic Gram-negative bacteria in many fields of healthcare.

  4. Activity of Ca2+-dependent neutral proteases in tissues of ground squirrel during hibernation and during self-warming after induced awakening.

    PubMed

    Nurmagomedova, P M; Abasova, M M; Emirbekov, E Z

    2011-09-01

    Cyclic changes in activity of Ca2+-dependent neutral protease occur during preparation for hibernation, with an increase in September and November and decrease in October and December. During hibernation proteolytic enzyme activity decreased, while during self-warming after induced awakening, the role of Ca2+-dependent processes in the tissues of ground squirrels increased according to the body temperature.

  5. Neutralization, by a monospecific Bothrops lanceolatus antivenom, of toxic activities induced by homologous and heterologous Bothírops snake venoms.

    PubMed

    Bogarín, G; Romero, M; Rojas, G; Lutsch, C; Casadamont, M; Lang, J; Otero, R; Gutiérrez, J M

    1999-03-01

    A monospecific Bothrops lanceolatus antivenom, currently used in Martinique, was tested for its efficacy in the neutralization of several toxic and enzymatic activities of the venoms of B. lanceolatus, B. atrox and B. asper. When tested by the i.p. route in mice, B. lanceolatus venom had an LD50 of 12.8 microg/g. In addition, it induced local tissue damage (hemorrhage, edema and myotoxicity) and showed indirect hemolytic activity, but was devoid of coagulant effect on human plasma in vitro and of defibrinating activity in mice. Antivenom was fully effective in the neutralization of lethal, hemorrhagic, edema-forming, myotoxic and indirect hemolytic effects of B. lanceolatus venom in assays involving preincubation of venom and antivenom. When tested against the venoms of B. asper and B. atrox, the antivenom completely neutralized the lethal, hemorrhagic, myotoxic and indirect hemolytic effects, and was partially effective in neutralizing edema-forming activity. In contrast, the antivenom was ineffective in the neutralization of in vitro coagulant and in vivo defibrinating effects induced by these two venoms. PMID:10080358

  6. Occurrence of a cytosolic neutral chitobiase activity involved in oligomannoside degradation: a study with Madin-Darby bovine kidney (MDBK) cells.

    PubMed

    Cacan, R; Dengremont, C; Labiau, O; Kmiécik, D; Mir, A M; Verbert, A

    1996-01-15

    Neutral oligomannosides possessing one GlcNAc (OS-Gn1) and two GlcNAc (Os-Gn2) at the reducing end have been reported to be released during the N-glycosylation process in various biological models. To investigate which enzyme is responsible for OS-Gn1 formation, we used the Madin-Darby bovine kidney (MDBK) cell line which exhibits neither lysosomal chitobiase nor endoglucosaminidase activities. However, these cells produced OS-Gn1 and we showed that a neutral chitobiase is responsible for the transformation of OS-Gn2 into OS-Gn1. Using streptolysin O-permeabilized MDBK cells, we demonstrated that this neutral chitobiase activity is located in the cytosolic compartment and is active on oligomannoside species released during the N-glycosylation process. PMID:8573098

  7. Occurrence of a cytosolic neutral chitobiase activity involved in oligomannoside degradation: a study with Madin-Darby bovine kidney (MDBK) cells.

    PubMed Central

    Cacan, R; Dengremont, C; Labiau, O; Kmiécik, D; Mir, A M; Verbert, A

    1996-01-01

    Neutral oligomannosides possessing one GlcNAc (OS-Gn1) and two GlcNAc (Os-Gn2) at the reducing end have been reported to be released during the N-glycosylation process in various biological models. To investigate which enzyme is responsible for OS-Gn1 formation, we used the Madin-Darby bovine kidney (MDBK) cell line which exhibits neither lysosomal chitobiase nor endoglucosaminidase activities. However, these cells produced OS-Gn1 and we showed that a neutral chitobiase is responsible for the transformation of OS-Gn2 into OS-Gn1. Using streptolysin O-permeabilized MDBK cells, we demonstrated that this neutral chitobiase activity is located in the cytosolic compartment and is active on oligomannoside species released during the N-glycosylation process. PMID:8573098

  8. Occurrence of a cytosolic neutral chitobiase activity involved in oligomannoside degradation: a study with Madin-Darby bovine kidney (MDBK) cells.

    PubMed

    Cacan, R; Dengremont, C; Labiau, O; Kmiécik, D; Mir, A M; Verbert, A

    1996-01-15

    Neutral oligomannosides possessing one GlcNAc (OS-Gn1) and two GlcNAc (Os-Gn2) at the reducing end have been reported to be released during the N-glycosylation process in various biological models. To investigate which enzyme is responsible for OS-Gn1 formation, we used the Madin-Darby bovine kidney (MDBK) cell line which exhibits neither lysosomal chitobiase nor endoglucosaminidase activities. However, these cells produced OS-Gn1 and we showed that a neutral chitobiase is responsible for the transformation of OS-Gn2 into OS-Gn1. Using streptolysin O-permeabilized MDBK cells, we demonstrated that this neutral chitobiase activity is located in the cytosolic compartment and is active on oligomannoside species released during the N-glycosylation process.

  9. Design and evaluation of a hybrid passive and active gravity neutral orthosis (GNO).

    PubMed

    Koo, Benjamin; Montes, Jacqueline; Gamarnik, Viktor; Yeager, Keith; Marra, Jonathan; Dunaway, Sally; Montgomery, Megan; De Vivo, Darryl C; Strauss, Nancy; Konofagou, Elisa; Kaufmann, Petra; Morrison, Barclay

    2009-01-01

    Neuromuscular diseases (NMD), including Spinal Muscular Atrophy (SMA) and Duchenne Muscular Dystrophy (DMD), result in progressive muscular weakness that often leaves patients functionally dependent on caregivers for many activities of daily living (ADL) such as eating, bathing, grooming (touching the face and head), reaching (grabbing for objects), and dressing. In severe cases, patients are unable to perform even the simplest of activities from exploring their 3D space to touching their own face. The ability to move and initiate age appropriate tasks, such as playing and exploration, are considered to be of vital importance to both their physical and cognitive development. Therefore, to improve quality of life and reduce dependence on caregivers in children and young adults with NMD, we designed, built and evaluated an assistive, active orthosis to support arm function. The goal of this project is the development and evaluation of a mechanical arm orthosis to both encourage and assist functional arm movement while providing the user a sense of independence and control over one's own body. PMID:19963513

  10. Effect of activation and preactivation on the mechanical behavior and neutral position of stainless steel and beta-titanium T-loops

    PubMed Central

    Moreira, Rui; Braga, Ana Cristina; Ferreira, Afonso Pinhão; Pollmann, Maria Cristina

    2015-01-01

    Objective To quantify, for each activation, the effect of preactivations of differing distribution and intensity on the neutral position of T-loops (7-mm height), specifically the horizontal force, moment to force (M/F) ratio, and load to deflection ratio. Methods A total 100 loops measuring 0.017 × 0.025 inches in cross-section were divided into two groups (n = 50 each) according to composition, either stainless steel or beta-titanium. The two groups were further divided into five subgroups, 10 loops each, corresponding to the five preactivations tested: preactivations with occlusal distribution (0°, 20°, and 40°), gingival distribution (20°), and occlusal-gingival distribution (40°). The loops were subjected to a total activation of 6-mm with 0.5-mm iterations. Statistical analysis was performed using comprised ANOVA and Bonferoni multiple comparison tests, with a significance level of 5%. Results The location and intensity of preactivation influenced the force intensity. For the M/F ratio, the highest value achieved without preactivation was lower than the height of the loop. Without preactivation, the M/F ratio increased with activation, while the opposite effect was observed with preactivation. The increase in the M/F ratio was greater when the preactivation distribution was partially or fully gingival. Conclusions Depending on the preactivation distribution, displacement of uprights is higher or lower than the activation, which is a factor to consider in clinical practice. PMID:26258066

  11. Identification of three competitive inhibitors for membrane-associated, Mg2+-dependent and neutral 60 kDa sphingomyelinase activity.

    PubMed

    Kim, Seok Kyun; Jung, Sang Mi; Ahn, Kyong Hoon; Jeon, Hyung Jun; Lee, Dong Hun; Jung, Kwang Mook; Jung, Sung Yun; Kim, Dae Kyong

    2005-08-01

    Methanol extracts of domestic plants of Korea were evaluated as a potential inhibitor of neutral pH optimum and membrane-associated 60 kDa sphingomyelinase (N-SMase) activity. In this study, we partially purified N-SMase from bovine brain membranes using ammonium sulfate. It was purified approximately 163-fold by the sequential use of DE52, Butyl-Toyopearl, DEAE-Cellulose, and Phenyl-5PW column chromatographies. The purified N-SMase activity was assayed in the presence of the plant extracts of three hundreds species. Based on the in vitro assay, three plant extracts significantly inhibited the N-SMase activity in a time- and concentration-dependent manner. To further examine the inhibitory pattern, a Dixon plot was constructed for each of the plant extracts. The extracts of Abies nephrolepis, Acer tegmentosum, and Ginkgo biloba revealed a competitive inhibition with the inhibition constant (Ki) of 11.9 microg/ mL, 9.4 microg/mL, and 12.9 microg/mL, respectively. These extracts also inhibited in a dose-dependent manner the production of ceramide induced by serum deprivation in human neuroblastoma cell line SH-SY5Y.

  12. Identification of minute damage in composite bridge structures equipped with fiber optic sensors using the location of neutral axis and finite element analysis

    NASA Astrophysics Data System (ADS)

    Li, Xi; Glisic, Branko

    2016-04-01

    By definition, the neutral axis of a loaded composite beam structure is the curve along which the section experiences zero bending strain. When no axial loading is present, the location of the neutral axis passes through the centroid of stiffness of the beam cross-section. In the presence of damage, the centroid of stiffness, as well as the neutral axis, shift from the healthy position. The concept of neutral axis can be widely applied to all beam-like structures. According to literature, a change in location of the neutral axis can be associated with damage in the corresponding cross-section. In this paper, the movement of neutral axis near locations of minute damage in a composite bridge structure was studied using finite element analysis and experimental results. The finite element model was developed based on a physical scale model of a composite simply-supported structure with controlled minute damage in the reinforced concrete deck. The structure was equipped with long-gauge fiber optic strain and temperature sensors at a healthy reference location as well as two locations of damage. A total of 12 strain sensors were installed during construction and used to monitor the structure during various loading events. This paper aims to explain previous experimental results which showed that the observed positions of neutral axis near damage locations were higher than the predicted healthy locations in some loading events. Analysis has shown that finite element analysis has potential to simulate and explain the physical behavior of the test structure.

  13. Neutral particle beam intensity controller

    DOEpatents

    Dagenhart, William K.

    1986-01-01

    A neutral beam intensity controller is provided for a neutral beam generator in which a neutral beam is established by accelerating ions from an ion source into a gas neutralizer. An amplitude modulated, rotating magnetic field is applied to the accelerated ion beam in the gas neutralizer to defocus the resultant neutral beam in a controlled manner to achieve intensity control of the neutral beam along the beam axis at constant beam energy. The rotating magnetic field alters the orbits of ions in the gas neutralizer before they are neutralized, thereby controlling the fraction of neutral particles transmitted out of the neutralizer along the central beam axis to a fusion device or the like. The altered path or defocused neutral particles are sprayed onto an actively cooled beam dump disposed perpendicular to the neutral beam axis and having a central open for passage of the focused beam at the central axis of the beamline. Virtually zero therough 100% intensity control is achieved by varying the magnetic field strength without altering the ion source beam intensity or its species yield.

  14. Theoretical study of chlordecone and surface groups interaction in an activated carbon model under acidic and neutral conditions.

    PubMed

    Gamboa-Carballo, Juan José; Melchor-Rodríguez, Kenia; Hernández-Valdés, Daniel; Enriquez-Victorero, Carlos; Montero-Alejo, Ana Lilian; Gaspard, Sarra; Jáuregui-Haza, Ulises Javier

    2016-04-01

    Activated carbons (ACs) are widely used in the purification of drinking water without almost any knowledge about the adsorption mechanisms of the persistent organic pollutants. Chlordecone (CLD, Kepone) is an organochlorinated synthetic compound that has been used mainly as agricultural insecticide. CLD has been identified and listed as a persistent organic pollutant by the Stockholm Convention. The selection of the best suited AC for this type of contaminants is mainly an empirical and costly process. A theoretical study of the influence of AC surface groups (SGs) on CLD adsorption is done in order to help understanding the process. This may provide a first selection criteria for the preparation of AC with suitable surface properties. A model of AC consisting of a seven membered ring graphene sheet (coronene) with a functional group on the edge was used to evaluate the influence of the SGs over the adsorption. Multiple Minima Hypersurface methodology (MMH) coupled with PM7 semiempirical Hamiltonian was employed in order to study the interactions of the chlordecone with SGs (hydroxyl and carboxyl) at acidic and neutral pH and different hydration conditions. Selected structures were re-optimized using CAM-B3LYP to achieve a well-defined electron density to characterize the interactions by the Quantum Theory of Atoms in Molecules approach. The deprotonated form of surface carboxyl and hydroxyl groups of AC models show the strongest interactions, suggesting a chemical adsorption. An increase in carboxylic SGs content is proposed to enhance CLD adsorption onto AC at neutral pH conditions. PMID:26945637

  15. Theoretical study of chlordecone and surface groups interaction in an activated carbon model under acidic and neutral conditions.

    PubMed

    Gamboa-Carballo, Juan José; Melchor-Rodríguez, Kenia; Hernández-Valdés, Daniel; Enriquez-Victorero, Carlos; Montero-Alejo, Ana Lilian; Gaspard, Sarra; Jáuregui-Haza, Ulises Javier

    2016-04-01

    Activated carbons (ACs) are widely used in the purification of drinking water without almost any knowledge about the adsorption mechanisms of the persistent organic pollutants. Chlordecone (CLD, Kepone) is an organochlorinated synthetic compound that has been used mainly as agricultural insecticide. CLD has been identified and listed as a persistent organic pollutant by the Stockholm Convention. The selection of the best suited AC for this type of contaminants is mainly an empirical and costly process. A theoretical study of the influence of AC surface groups (SGs) on CLD adsorption is done in order to help understanding the process. This may provide a first selection criteria for the preparation of AC with suitable surface properties. A model of AC consisting of a seven membered ring graphene sheet (coronene) with a functional group on the edge was used to evaluate the influence of the SGs over the adsorption. Multiple Minima Hypersurface methodology (MMH) coupled with PM7 semiempirical Hamiltonian was employed in order to study the interactions of the chlordecone with SGs (hydroxyl and carboxyl) at acidic and neutral pH and different hydration conditions. Selected structures were re-optimized using CAM-B3LYP to achieve a well-defined electron density to characterize the interactions by the Quantum Theory of Atoms in Molecules approach. The deprotonated form of surface carboxyl and hydroxyl groups of AC models show the strongest interactions, suggesting a chemical adsorption. An increase in carboxylic SGs content is proposed to enhance CLD adsorption onto AC at neutral pH conditions.

  16. The simultaneous release by bone explants in culture and the parallel activation of procollagenase and of a latent neutral proteinase that degrades cartilage proteoglycans and denatured collagen.

    PubMed Central

    Vaes, G; Eeckhout, Y; Lenaers-Claeys, G; François-Gillet, C; Druetz, J E

    1978-01-01

    1. A latent neutral proteinase was found in culture media of mouse bone explants. Its accumulation during the cultures is closely parallel to that of procollagenase; both require the presence of heparin in the media. 2. Latent neutral proteinase was activated by several treatments of the media known to activate procollagenase, such as limited proteolysis by trypsin, chymotrypsin, plasmin or kallikrein, dialysis against 3 M-NaSCN at 4 degrees C and prolonged preincubation at 25 degrees C. Its activation often followed that of the procollagenase present in the same media. 3. Activation of neutral proteinase (as does that of procollagenase) by trypsin or plasmin involved two successive steps: the activation of a latent endogenous activator present in the media followed by the activation of neutral proteinase itself by that activator. 4. The proteinase degrades cartilage proteoglycans, denatured collagen (Azocoll) and casein at neutral pH; it is inhibited by EDTA, cysteine or serum. Collagenase is not inhibited by casein or Azocoll and is less resistant to heat or to trypsin than is the proteinase. Partial separation of the two enzymes was achieved by gel filtration of the media but not by fractional (NH4)2SO4 precipitation, by ion exchange or by affinity chromatography on Sepharose-collagen. These fractionations did not activate latent enzymes. 5. Trypsin activation decreases the molecular weight of both latent enzymes (60 000-70 000) by 20 000-30 000, as determined by gel filtration of media after removal of heparin. 6. The latency of both enzymes could be due either to a zymogen or to an enzyme-inhibitor complex. A thermostable inhibitor of both enzymes was found in some media. However, combinations of either enzyme with that inhibitor were not reactivated by trypsin, indicating that this inhibitor is unlikely to be the cause of the latency. PMID:208518

  17. Mass analysis of neutral particles and ions released during electrical breakdowns on spacecraft surfaces

    NASA Technical Reports Server (NTRS)

    Kendall, B. R. F.

    1985-01-01

    Charged-particle fluxes from breakdown events were studied. Methods to measure mass spectra and total emitted flux of neutral particles were developed. The design and construction of the specialized mass spectrometer was completed. Electrical breakdowns were initiated by a movable blunt contact touching the insulating surface. The contact discharge apparatus was used for final development of two different high-speed recording systems and for measurements of the composition of the materials given off by the discharge. It was shown that intense instantaneous fluxes of neutral particles were released from the sites of electrical breakdown events. A laser micropulse mass analyzer showed that visible discoloration at breakdown sites were correllated with the presence of iron on the polymer side of the film, presumably caused by punch-through to the Inconel backing. Kapton samples irradiated by an oxygen ion beam were tested. The irradiated samples were free of surface hydrocarbon contamination but otherwise behaved in the same way as the Kapton samples tested earlier. Only the two samples exposed to oxygen ion bombardment were relatively clean. This indicates an additional variable that should be considered when testing spacecraft materials in the laboratory.

  18. Analysis of immobilized artificial membrane retention factors for both neutral and ionic species.

    PubMed

    Abraham, Michael H; Acree, William E; Fahr, Alfred; Liu, Xiangli

    2013-07-12

    Retention data on an immobilised artificial membrane have been taken from the work of Li et al. and from Liu et al., and have been correlated with a set of descriptors that includes descriptors for ionized species, that is anions from deprotonated acids and cations from protonated bases. Log k(IAM) values can be predicted for acids or bases that are partially ionized at the experimental pH and log k(IAM) values for acids and bases can be predicted as a function of the fraction present as the ionized species, equivalent to prediction as a function of pH. It is shown that anions reduce the value of log k(IAM) by about 1.1 log units but that cations have almost no effect by comparison to the neutral species. By comparison to non-polar solvents, carboxylate anions and protonated base cations are considerably stabilized by both water and the IAM phase to about 6-8 log units and so the rather small influence of anions (-1.1 log unit) and cations (-0.1 log unit) on log k(IAM) is due to substantial cancellation of these stabilization effects. Indeed, the effect of change of phase from water to IAM on the neutral species is at least as large as the effect of change of phase on the ionic species.

  19. Exosomes neutralize synaptic-plasticity-disrupting activity of Aβ assemblies in vivo

    PubMed Central

    2013-01-01

    Background Exosomes, small extracellular vesicles of endosomal origin, have been suggested to be involved in both the metabolism and aggregation of Alzheimer’s disease (AD)-associated amyloid β-protein (Aβ). Despite their ubiquitous presence and the inclusion of components which can potentially interact with Aβ, the role of exosomes in regulating synaptic dysfunction induced by Aβ has not been explored. Results We here provide in vivo evidence that exosomes derived from N2a cells or human cerebrospinal fluid can abrogate the synaptic-plasticity-disrupting activity of both synthetic and AD brain-derived Aβ. Mechanistically, this effect involves sequestration of synaptotoxic Aβ assemblies by exosomal surface proteins such as PrPC rather than Aβ proteolysis. Conclusions These data suggest that exosomes can counteract the inhibitory action of Aβ, which contributes to perpetual capability for synaptic plasticity. PMID:24284042

  20. Interferon α kinoid induces neutralizing anti-interferon α antibodies that decrease the expression of interferon-induced and B cell activation associated transcripts: analysis of extended follow-up data from the interferon α kinoid phase I/II study

    PubMed Central

    Ducreux, Julie; Houssiau, Frédéric A.; Vandepapelière, Pierre; Jorgensen, Christian; Lazaro, Estibaliz; Spertini, François; Colaone, Fabien; Roucairol, Camille; Laborie, Marion; Croughs, Thérèse; Lauwerys, Bernard R.

    2016-01-01

    Objective. IFN α Kinoid (IFN-K) is a therapeutic vaccine composed of IFNα2b coupled to a carrier protein. In a phase I/II placebo-controlled trial, we observed that IFN-K significantly decreases the IFN gene signature in whole blood RNA samples from SLE patients. Here, we analysed extended follow-up data from IFN-K-treated patients, in order to evaluate persistence of neutralizing anti-IFNα Abs antibodies (Abs), and gene expression profiling. Methods. Serum and whole blood RNA samples were obtained in IFN-K-treated patients included in the follow-up study, in order to determine binding and neutralizing anti-IFNα Ab titres, and perform high-throughput transcriptomic studies. Results. Neutralization studies of 13 IFNα subtypes demonstrated the polyclonal nature of the Ab response induced by IFN-K. Follow-up analyses in six patients confirmed a significant correlation between neutralizing anti-IFNα Ab titres and decrease in IFN scores compared to baseline. These analyses also revealed an inhibitory effect of IFNα blockade on the expression of B cell associated transcripts. Conclusions. IFN-K induces a polyclonal anti-IFNα response that decreases IFN- and B cell-associated transcripts. Trial registration: ClinicalTrials.gov, clinicaltrials.gov, NCT01058343 PMID:27354683

  1. HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity.

    PubMed

    Matoba, Nobuyuki; Husk, Adam S; Barnett, Brian W; Pickel, Michelle M; Arntzen, Charles J; Montefiori, David C; Takahashi, Atsushi; Tanno, Kazunobu; Omura, Satoshi; Cao, Huyen; Mooney, Jason P; Hanson, Carl V; Tanaka, Haruo

    2010-01-01

    The development of a topical microbicide blocking the sexual transmission of HIV-1 is urgently needed to control the global HIV/AIDS pandemic. The actinomycete-derived lectin actinohivin (AH) is highly specific to a cluster of high-mannose-type glycans uniquely found on the viral envelope (Env). Here, we evaluated AH's candidacy toward a microbicide in terms of in vitro anti-HIV-1 activity, potential side effects, and recombinant producibility. Two validated assay systems based on human peripheral blood mononuclear cell (hPBMC) infection with primary isolates and TZM-bl cell infection with Env-pseudotyped viruses were employed to characterize AH's anti-HIV-1 activity. In hPMBCs, AH exhibited nanomolar neutralizing activity against primary viruses with diverse cellular tropisms, but did not cause mitogenicity or cytotoxicity that are often associated with other anti-HIV lectins. In the TZM-bl-based assay, AH showed broad anti-HIV-1 activity against clinically-relevant, mucosally transmitting strains of clades B and C. By contrast, clade A viruses showed strong resistance to AH. Correlation analysis suggested that HIV-1's AH susceptibility is significantly linked to the N-glycans at the Env C2 and V4 regions. For recombinant (r)AH expression, we evaluated a tobacco mosaic virus-based system in Nicotiana benthamiana plants as a means to facilitate molecular engineering and cost-effective mass production. Biochemical analysis and an Env-mediated syncytium formation assay demonstrated high-level expression of functional rAH within six days. Taken together, our study revealed AH's cross-clade anti-HIV-1 activity, apparent lack of side effects common to lectins, and robust producibility using plant biotechnology. These findings justify further efforts to develop rAH toward a candidate HIV-1 microbicide. PMID:20559567

  2. Simulation of Cl− Secretion in Epithelial Tissues: New Methodology Estimating Activity of Electro-Neutral Cl− Transporter

    PubMed Central

    Sasamoto, Kouhei; Niisato, Naomi; Taruno, Akiyuki; Marunaka, Yoshinori

    2015-01-01

    Transcellular Cl− secretion is, in general, mediated by two steps; (1) the entry step of Cl− into the cytosolic space from the basolateral space across the basolateral membrane by Cl− transporters, such as Na+-K+-2Cl− cotransporter (NKCC1, an isoform of NKCC), and (2) the releasing step of Cl− from the cytosolic space into the luminal (air) space across the apical membrane via Cl− channels, such as cystic fibrosis transmembrane conductance regulator (CFTR) Cl− channel. Transcellular Cl− secretion has been characterized by using various experimental techniques. For example, measurements of short-circuit currents in the Ussing chamber and patch clamp techniques provide us information on transepithelial ion movements via transcellular pathway, transepithelial conductance, activity (open probability) of single channel, and whole cell currents. Although many investigators have tried to clarify roles of Cl− channels and transporters located at the apical and basolateral membranes in transcellular Cl− secretion, it is still unclear how Cl− channels/transporters contribute to transcellular Cl− secretion and are regulated by various stimuli such as Ca2+ and cAMP. In the present study, we simulate transcellular Cl− secretion using mathematical models combined with electrophysiological measurements, providing information on contribution of Cl− channels/transporters to transcellular Cl− secretion, activity of electro-neutral ion transporters and how Cl− channels/transporters are regulated. PMID:26779025

  3. The neuromuscular activity of Micrurus pyrrhocryptus venom and its neutralization by commercial and specific coral snake antivenoms

    PubMed Central

    Camargo, Thiago Magalhães; de Roodt, Adolfo Rafael; da Cruz-Höfling, Maria Alice; Rodrigues-Simioni, Léa

    2011-01-01

    The neuromuscular activity ofMicrurus pyrrochryptus venom was studied in chick biventer cervicis (BC) and mouse phrenic nerve-diaphragm (PND) preparations. The venom (0.5-50μg/ml) caused irreversible, time- and concentration-dependent blockade, with BC being more sensitive than PND (50% blockade with 10μg/ml in 22±;3min and 62±4min, respectively; mean±SEM, n=6; p<0.05). In BC preparations, venom (0.5μg/ml) progressively abolished ACh-induced contractures, whereas contractures to exogenous KCl and muscle twitches in curarized preparations were unaffected. The venom neither altered creatine kinase release (venom: 25.8±1.75IU/l vs control: 24.3±2.2IU/l, n=6, after 120min), nor it caused significant muscle damage (50μg of venom/ml vs control: 3.5±0.8% vs 1.1±0.7% for PND; 4.3±1.5% vs 1.2±0.5% for BC, n=5). The venom had low PLA2 activity. Neurotoxicity was effectively neutralized by commercial Micrurus antivenom and specific antivenom. These findings indicate that M. pyrrhocryptus venom acts postsynaptically on nicotinic receptors, with no significant myotoxicity. PMID:21858249

  4. Bench-scale study of active mine water treatment using cement kiln dust (CKD) as a neutralization agent.

    PubMed

    Mackie, Allison L; Walsh, Margaret E

    2012-02-01

    The overall objective of this study was to investigate the potential impact on settled water quality of using cement kiln dust (CKD), a waste by-product, to replace quicklime in the active treatment of acidic mine water. Bench-scale experiments were conducted to evaluate the treatment performance of calcium hydroxide (Ca(OH)(2)) slurries generated using four different CKD samples compared to a control treatment with quicklime (CaO) in terms of reducing acidity and metals concentrations in acid mine drainage (AMD) samples taken from the effluent of a lead/zinc mine in Atlantic Canada. Results of the study showed that all of the CKD samples evaluated were capable of achieving greater than 97% removal of total zinc and iron. The amount of solid alkaline material required to achieve pH targets required for neutralization of the AMD was found to be higher for treatment with the CKD slurries compared to the quicklime slurry control experiments, and varied linearly with the free lime content of the CKD. The results of this study also showed that a potential benefit of treating mine water with CKD could be reduced settled sludge volumes generated in the active treatment process, and further research into the characteristics of the sludge generated from the use of CKD-generated calcium hydroxide slurries is recommended.

  5. The neuromuscular activity of Micrurus pyrrhocryptus venom and its neutralization by commercial and specific coral snake antivenoms.

    PubMed

    Camargo, Thiago Magalhães; de Roodt, Adolfo Rafael; da Cruz-Höfling, Maria Alice; Rodrigues-Simioni, Léa

    2011-01-01

    The neuromuscular activity ofMicrurus pyrrochryptus venom was studied in chick biventer cervicis (BC) and mouse phrenic nerve-diaphragm (PND) preparations. The venom (0.5-50μg/ml) caused irreversible, time- and concentration-dependent blockade, with BC being more sensitive than PND (50% blockade with 10μg/ml in 22±;3min and 62±4min, respectively; mean±SEM, n=6; p<0.05). In BC preparations, venom (0.5μg/ml) progressively abolished ACh-induced contractures, whereas contractures to exogenous KCl and muscle twitches in curarized preparations were unaffected. The venom neither altered creatine kinase release (venom: 25.8±1.75IU/l vs control: 24.3±2.2IU/l, n=6, after 120min), nor it caused significant muscle damage (50μg of venom/ml vs control: 3.5±0.8% vs 1.1±0.7% for PND; 4.3±1.5% vs 1.2±0.5% for BC, n=5). The venom had low PLA(2) activity. Neurotoxicity was effectively neutralized by commercial Micrurus antivenom and specific antivenom. These findings indicate that M. pyrrhocryptus venom acts postsynaptically on nicotinic receptors, with no significant myotoxicity. PMID:21858249

  6. Neutralization of the haemorrhagic activities of viperine snake venoms and venom metalloproteinases using synthetic peptide inhibitors and chelators.

    PubMed

    Howes, J-M; Theakston, R D G; Laing, G D

    2007-04-01

    Envenoming by the West African saw-scaled viper, Echis ocellatus resembles that of most vipers, in that it results in local blistering, necrosis and sometimes life-threatening systemic haemorrhage. While effective against systemic envenoming, current antivenoms have little or no effect against local tissue damage. The major mediators of local venom pathology are the zinc-dependant snake venom metalloproteinases (SVMPs). The high degree of structural and functional homology between SVMPs and their mammalian relatives the matrix metalloproteinases (MMPs) suggests that substrate/inhibitor interactions between these subfamilies are likely to be analogous. In this study, four recently developed MMP inhibitors (MMPIs) (Marimastat, AG-3340, CGS-270 23A and Bay-12 9566) are evaluated in addition to three metal ion chelators (EDTA, TPEN and BAPTA) for their ability to inhibit the haemorrhagic activities of the medically important E. ocellatus venom and one of its haemorrhagic SVMPs, EoVMP2. As expected, the metal ion chelators significantly inhibited the haemorrhagic activities of both whole E. ocellatus venom and EoVMP2, while the synthetic MMPIs show more variation in their efficacies. These variations suggest that individual MMPIs show specificity towards SVMPs and that their application to the neutralization of local haemorrhage may require a synthetic MMPI mixture, ensuring that a close structural component for each SVMP is represented. PMID:17196631

  7. Expression patterns, activities and carbohydrate-metabolizing regulation of sucrose phosphate synthase, sucrose synthase and neutral invertase in pineapple fruit during development and ripening.

    PubMed

    Zhang, Xiu-Mei; Wang, Wei; Du, Li-Qing; Xie, Jiang-Hui; Yao, Yan-Li; Sun, Guang-Ming

    2012-01-01

    Differences in carbohydrate contents and metabolizing-enzyme activities were monitored in apical, medial, basal and core sections of pineapple (Ananas comosus cv. Comte de paris) during fruit development and ripening. Fructose and glucose of various sections in nearly equal amounts were the predominant sugars in the fruitlets, and had obvious differences until the fruit matured. The large rise of sucrose/hexose was accompanied by dramatic changes in sucrose phosphate synthase (SPS) and sucrose synthase (SuSy) activities. By contrast, neutral invertase (NI) activity may provide a mechanism to increase fruit sink strength by increasing hexose concentrations. Furthermore, two cDNAs of Ac-sps (accession no. GQ996582) and Ac-ni (accession no. GQ996581) were first isolated from pineapple fruits utilizing conserved amino-acid sequences. Homology alignment reveals that the amino acid sequences contain some conserved function domains. Transcription expression analysis of Ac-sps, Ac-susy and Ac-ni also indicated distinct patterns related to sugar accumulation and composition of pineapple fruits. It suggests that differential expressions of multiple gene families are necessary for sugar metabolism in various parts and developmental stages of pineapple fruit. A cycle of sucrose breakdown in the cytosol of sink tissues could be mediated through both Ac-SuSy and Ac-NI, and Ac-NI could be involved in regulating crucial steps by generating sugar signals to the cells in a temporally and spatially restricted fashion.

  8. Mass analysis of neutral particles and ions released during electrical breakdowns on spacecraft surfaces

    NASA Technical Reports Server (NTRS)

    Kendall, B. R. F.

    1983-01-01

    A specialized spectrometer was designed and developed to measure the mass and velocity distributions of neutral particles (molecules and molecular clusters) released from metal-backed Teflon and Kapton films. Promising results were obtained with an insulation breakdown initiation system based on a moveable contact touching the insulated surfaces. A variable energy, high voltage pulse is applied to the contact. The resulting surface damage sites can be made similar in size and shape to those produced by a high voltage electron beam system operating at similar discharge energies. The point discharge apparatus was used for final development of several high speed recording systems and for measurements of the composition of the materials given off by the discharge. Results with this apparatus show evolution of large amounts of fluorocarbon fragments from discharge through Teflon FEP, while discharges through Kapton produce mainly very light hydrocarbon fragments at masses below about 80 a.m.u.

  9. Transition Probabilities for Neutral Cerium from Boltzmann Analysis of Fourier Transform Spectra

    NASA Astrophysics Data System (ADS)

    Nitz, D. E.; Curry, J. J.; Buuck, M. J.; Mitchell, N. P.; Demann, A. D.; Shull, W. E.

    2012-06-01

    The recent availability of a large set of absolute transition probabilities for neutral cerium (Lawler et. al., J. Phys. B 43, 85701 (2010)) makes it possible to investigate the relative populations of the upper levels of these lines in radiometrically-calibrated spectra. In cases where these populations can be characterized by a single effective Boltzmann temperature, applying this temperature enables one to determine additional absolute transition probabilities for observable decay branches of nearby levels. While not as accurate as measurements based on branching fractions and lifetimes, the method can be applied to levels whose lifetimes are not known and does not require accounting for all of the branches. We are analyzing Fourier Transform spectra from NIST and from the National Solar Observatory data archive at Kitt Peak via this technique, seeking to increase the set of known transition probabilities for Ce I by a factor of 2-3. A summary of results obtained to date will be presented.

  10. Analysis of secondary particle behavior in multiaperture, multigrid accelerator for the ITER neutral beam injector.

    PubMed

    Mizuno, T; Taniguchi, M; Kashiwagi, M; Umeda, N; Tobari, H; Watanabe, K; Dairaku, M; Sakamoto, K; Inoue, T

    2010-02-01

    Heat load on acceleration grids by secondary particles such as electrons, neutrals, and positive ions, is a key issue for long pulse acceleration of negative ion beams. Complicated behaviors of the secondary particles in multiaperture, multigrid (MAMuG) accelerator have been analyzed using electrostatic accelerator Monte Carlo code. The analytical result is compared to experimental one obtained in a long pulse operation of a MeV accelerator, of which second acceleration grid (A2G) was removed for simplification of structure. The analytical results show that relatively high heat load on the third acceleration grid (A3G) since stripped electrons were deposited mainly on A3G. This heat load on the A3G can be suppressed by installing the A2G. Thus, capability of MAMuG accelerator is demonstrated for suppression of heat load due to secondary particles by the intermediate grids.

  11. Ebola virus-like particles produced in insect cells exhibit dendritic cell stimulating activity and induce neutralizing antibodies

    SciTech Connect

    Ye Ling; Lin Jianguo; Sun Yuliang; Bennouna, Soumaya; Lo, Michael; Wu Qingyang; Bu Zhigao; Pulendran, Bali; Compans, Richard W. . E-mail: compans@microbio.emory.edu; Yang Chinglai . E-mail: chyang@emory.edu

    2006-08-01

    Recombinant baculoviruses (rBV) expressing Ebola virus VP40 (rBV-VP40) or GP (rBV-GP) proteins were generated. Infection of Sf9 insect cells by rBV-VP40 led to assembly and budding of filamentous particles from the cell surface as shown by electron microscopy. Ebola virus-like particles (VLPs) were produced by coinfection of Sf9 cells with rBV-VP40 and rBV-GP, and incorporation of Ebola GP into VLPs was demonstrated by SDS-PAGE and Western blot analysis. Recombinant baculovirus infection of insect cells yielded high levels of VLPs, which were shown to stimulate cytokine secretion from human dendritic cells similar to VLPs produced in mammalian cells. The immunogenicity of Ebola VLPs produced in insect cells was evaluated by immunization of mice. Analysis of antibody responses showed that most of the GP-specific antibodies were of the IgG2a subtype, while no significant level of IgG1 subtype antibodies specific for GP was induced, indicating the induction of a Th1-biased immune response. Furthermore, sera from Ebola VLP immunized mice were able to block infection by Ebola GP pseudotyped HIV virus in a single round infection assay, indicating that a neutralizing antibody against the Ebola GP protein was induced. These results show that production of Ebola VLPs in insect cells using recombinant baculoviruses represents a promising approach for vaccine development against Ebola virus infection.

  12. Decanting of Neutralized H-Canyon Unirradiated Nuclear Material High Activity Waste Streams

    SciTech Connect

    BRONIKOWSKI, MICHAELG.

    2004-08-05

    An option to dispose of the High Activity Waste (HAW) stream from the processing of unirradiated materials directly to Saltstone is being evaluated to conserve High Level Waste (HLW) tank farm space and to reduce the future production of HLW glass logs. To meet the Saltstone Waste Acceptance Criteria (WAC), decanting the supernate from precipitated solids was proposed to reduce mercury and radionuclide levels in the waste. Only the caustic supernate will then be sent to Saltstone. Verification that the Saltstone WAC will be met has involved a series of laboratory studies using surrogate and actual HAW solutions from H-Canyon. The initial experiment involved addition of sodium hydroxide (NaOH) to a surrogate HAW test solution and subsequent decanting of the supernate away from the precipitated solids. The chemical composition of the surrogate solution was based on a composition defined from analyses of actual HAW solutions generated during dissolution of unirradiated nuclear materials in H-Canyon [1]. Results from testing the surrogate HAW solution were reported in Reference [2]. Information obtained from the surrogate test solution study was used to define additional experiments on actual HAW solutions obtained from H-Canyon. These experiments were conducted with samples from three different batches of HAW solutions. The first and third HAW samples (HAW No.1 and HAW No.3 solutions) contained the centrifuge filter cake material from a gelatin strike that is periodically added to the waste stream. The second HAW sample (HAW No.2 solution) did not contain filter cake material. Monosodium titanate (MST) was added to the HAW No.2 and HAW No.3 solutions after addition of NaOH was complete and before the settling step. The addition of MST was to improve the decontamination of alpha and beta emitters (primarily plutonium and strontium) from the supernate. The addition of excess NaOH and the addition of MST were expected to result in sufficient alpha and beta

  13. Evolutionary neutrality of mtDNA introgression: evidence from complete mitogenome analysis in roe deer.

    PubMed

    Matosiuk, M; Sheremetyeva, I N; Sheremetyev, I S; Saveljev, A P; Borkowska, A

    2014-11-01

    Introgressive hybridization offers a unique platform for studying the molecular basis of natural selection acting on mitogenomes. Most of the mtDNA protein-coding genes are extremely conserved; however, some of the observed variations have potentially adaptive significance. Here, we evaluated whether the evolution of mtDNA in closely related roe deer species affected by widespread mtDNA introgression is neutral or adaptive. We characterized and compared 16 complete mitogenomes of European (Capreolus capreolus) and Siberian (C. pygargus) roe deer, including four of Siberian origin introgressed into European species. The average sequence divergence of species-specific lineages was estimated at 2.8% and varied across gene classes. Only 21 of 315 fixed differences identified in protein-coding genes represented nonsynonymous changes. Only three of them were determined to have arisen in the C. pygargus lineage since the time to the most recent common ancestor (TMRCA) of both Capreolus species, reflecting a decelerated evolutionary ratio. The almost four-fold higher dN /dS ratio described for the European roe deer lineage is constrained by overall purifying selection, especially pronounced in the ND4 and ND5 genes. We suggest that the highly divergent C. capreolus lineage could have maintained a capability for genomic incorporation of the well-preserved and almost ancestral type of mtDNA present in C. pygargus. Our analyses did not indicate any signs of positive selection for Siberian roe deer mtDNA, suggesting that the present widespread introgression is evolutionarily neutral.

  14. Direct analysis of sialylated or sulfated glycosphingolipids and other polar and neutral lipids using TLC-MS interfaces.

    PubMed

    Park, Hyejung; Zhou, Ying; Costello, Catherine E

    2014-04-01

    Gangliosides and sulfatides (STs) are acidic glycosphingolipids (GSLs) that have one or more sialic acids or sulfate substituents, in addition to neutral sugars, attached to the C-1 hydroxyl group of the ceramide long chain base. TLC is a widely employed and convenient technique for separation and characterization of GSLs. When TLC is directly coupled to MS, it provides both the molecular mass and structural information without further purification. Here, after development of the TLC plates, the structural analyses of acidic GSLs, including gangliosides and STs, were investigated using the liquid extraction surface analysis (LESA™) and CAMAG TLC-MS interfaces coupled to an ESI QSTAR Pulsar i quadrupole orthogonal TOF mass spectrometer. Coupling TLC with ESI-MS allowed the acquisition of high resolution mass spectra of the acidic GSLs with high sensitivity and mass accuracy, without the loss of sialic acid residues that frequently occurs during low-pressure MALDI MS. These systems were then applied to the analysis of total lipid extracts from bovine brain. This allowed profiling of many different lipid classes, not only gangliosides and STs, but also SMs, neutral GSLs, and phospholipids. PMID:24482490

  15. Variation of acharan sulfate and monosaccharide composition and analysis of neutral N-glycans in African giant snail (Achatina fulica)

    PubMed Central

    Park, Youmie; Zhang, Zhenqing; Laremore, Tatiana N.; Li, Boyangzi; Sim, Joon-Soo; Im, A-Rang; Ahn, Mi Young

    2009-01-01

    Acharan sulfate content from African giant snail (Achatina fulica) was compared in eggs and snails of different ages. Acharan sulfate was not found in egg. Acharan sulfate disaccharide →4)-α-d-GlcNpAc (1→4)-α-l-IdoAp2S(1→, analyzed by SAX (strong-anion exchange)–HPLC was observed soon after hatching and increases as the snails grow. Monosaccharide compositional analysis showed that mole % of glucosamine, a major monosaccharide of acharan sulfate, increased with age while mole % of galactose decreased with age. These results suggest that galactans represent a major energy source during development, while acharan sulfate appearing immediately after hatching, is essential for the snail growth. The structures of neutral N-glycans released from eggs by peptide N-glycosidase F (PNGase F), were next elucidated using ESI-MS/MS, MALDI-MS/MS, enzyme digestion, and monosaccharide composition analysis. Three types of neutral N-glycan structures were observed, truncated (Hex2–4-Hex-NAc2), high mannose (Hex5–9-HexNAc2), and complex (Hex3-HexNAc2–10) types. None showed core fucosylation. PMID:18670878

  16. Neutralizing activities of human immunoglobulin derived from donors in Japan against mosquito-borne flaviviruses, Japanese encephalitis virus, West Nile virus, and dengue virus

    PubMed Central

    Yunoki, Mikihiro; Kurosu, Takeshi; Koketsu, Ritsuko Kubota; Takahashi, Kazuo; Okuno, Yoshinobu; Ikuta, Kazuyoshi

    2016-01-01

    Japanese encephalitis virus (JEV), West Nile virus (WNV), and dengue virus (DenV) are causal agents of Japanese encephalitis, West Nile fever, and dengue fever, respectively. JEV is considered to be indigenized and widespread in Japan, whereas WNV and DenV are not indigenized in Japan. Globulin products seem to reflect the status of the donor population according to antivirus neutralization activity. However, the anti-JEV, -WNV, and -DenV neutralization activities of globulin products derived from donors in Japan have not been clarified. Furthermore, potential candidates for the development of an effective immunotherapeutic drug for encephalitis caused by JEV, WNV, or DenV have also not been identified. Therefore, the aim of this study was to determine the overall status of the donor population in Japan based on globulin products by evaluating anti-JEV, -WNV, and -DenV neutralizing activities of intravenous immunoglobulin. Overall, intravenous immunoglobulin products showed stable neutralizing activity against JEV but showed no or only weak activity against WNV or DenV. These results suggest that the epidemiological level against WNV and DenV in the donor population of Japan is still low, suggesting that these viruses are not yet indigenized. In addition, JEV vaccinations and/or infections in the donor population do not induce a cross-reactive antibody against WNV. PMID:27462140

  17. Neutralizing activities of human immunoglobulin derived from donors in Japan against mosquito-borne flaviviruses, Japanese encephalitis virus, West Nile virus, and dengue virus.

    PubMed

    Yunoki, Mikihiro; Kurosu, Takeshi; Koketsu, Ritsuko Kubota; Takahashi, Kazuo; Okuno, Yoshinobu; Ikuta, Kazuyoshi

    2016-01-01

    Japanese encephalitis virus (JEV), West Nile virus (WNV), and dengue virus (DenV) are causal agents of Japanese encephalitis, West Nile fever, and dengue fever, respectively. JEV is considered to be indigenized and widespread in Japan, whereas WNV and DenV are not indigenized in Japan. Globulin products seem to reflect the status of the donor population according to antivirus neutralization activity. However, the anti-JEV, -WNV, and -DenV neutralization activities of globulin products derived from donors in Japan have not been clarified. Furthermore, potential candidates for the development of an effective immunotherapeutic drug for encephalitis caused by JEV, WNV, or DenV have also not been identified. Therefore, the aim of this study was to determine the overall status of the donor population in Japan based on globulin products by evaluating anti-JEV, -WNV, and -DenV neutralizing activities of intravenous immunoglobulin. Overall, intravenous immunoglobulin products showed stable neutralizing activity against JEV but showed no or only weak activity against WNV or DenV. These results suggest that the epidemiological level against WNV and DenV in the donor population of Japan is still low, suggesting that these viruses are not yet indigenized. In addition, JEV vaccinations and/or infections in the donor population do not induce a cross-reactive antibody against WNV. PMID:27462140

  18. Secondary Neutral Mass Spectrometry: The application of laser post ionization to trace surface analysis in semiconductor materials. [Fe on O-implanted Si

    SciTech Connect

    Pellin, M.J.; Young, C.E.; Calaway, W.F.; Burnett, J.W.; Gruen, D.M.

    1988-01-01

    SNMS uses the secondary neutral sputtered fraction, and can increase both yield and quantitative analysis. In this paper, lasers are used to ionize and then detect the sputtered neutral particles. This method is used with oxygen ion implantation to study Fe on Si substrates. In this way, Fe could be detected at the 500 ppT level in a single monolayer. 34 refs., 6 figs. (DLC)

  19. Hyperimmune Bovine Colostrum as a Low-Cost, Large-Scale Source of Antibodies with Broad Neutralizing Activity for HIV-1 Envelope with Potential Use in Microbicides

    PubMed Central

    Kramski, Marit; Center, Rob J.; Wheatley, Adam K.; Jacobson, Jonathan C.; Alexander, Marina R.; Rawlin, Grant

    2012-01-01

    Bovine colostrum (first milk) contains very high concentrations of IgG, and on average 1 kg (500 g/liter) of IgG can be harvested from each immunized cow immediately after calving. We used a modified vaccination strategy together with established production systems from the dairy food industry for the large-scale manufacture of broadly neutralizing HIV-1 IgG. This approach provides a low-cost mucosal HIV preventive agent potentially suitable for a topical microbicide. Four cows were vaccinated pre- and/or postconception with recombinant HIV-1 gp140 envelope (Env) oligomers of clade B or A, B, and C. Colostrum and purified colostrum IgG were assessed for cross-clade binding and neutralization against a panel of 27 Env-pseudotyped reporter viruses. Vaccination elicited high anti-gp140 IgG titers in serum and colostrum with reciprocal endpoint titers of up to 1 × 105. While nonimmune colostrum showed some intrinsic neutralizing activity, colostrum from 2 cows receiving a longer-duration vaccination regimen demonstrated broad HIV-1-neutralizing activity. Colostrum-purified polyclonal IgG retained gp140 reactivity and neutralization activity and blocked the binding of the b12 monoclonal antibody to gp140, showing specificity for the CD4 binding site. Colostrum-derived anti-HIV antibodies offer a cost-effective option for preparing the substantial quantities of broadly neutralizing antibodies that would be needed in a low-cost topical combination HIV-1 microbicide. PMID:22664963

  20. Hyperimmune bovine colostrum as a low-cost, large-scale source of antibodies with broad neutralizing activity for HIV-1 envelope with potential use in microbicides.

    PubMed

    Kramski, Marit; Center, Rob J; Wheatley, Adam K; Jacobson, Jonathan C; Alexander, Marina R; Rawlin, Grant; Purcell, Damian F J

    2012-08-01

    Bovine colostrum (first milk) contains very high concentrations of IgG, and on average 1 kg (500 g/liter) of IgG can be harvested from each immunized cow immediately after calving. We used a modified vaccination strategy together with established production systems from the dairy food industry for the large-scale manufacture of broadly neutralizing HIV-1 IgG. This approach provides a low-cost mucosal HIV preventive agent potentially suitable for a topical microbicide. Four cows were vaccinated pre- and/or postconception with recombinant HIV-1 gp140 envelope (Env) oligomers of clade B or A, B, and C. Colostrum and purified colostrum IgG were assessed for cross-clade binding and neutralization against a panel of 27 Env-pseudotyped reporter viruses. Vaccination elicited high anti-gp140 IgG titers in serum and colostrum with reciprocal endpoint titers of up to 1 × 10(5). While nonimmune colostrum showed some intrinsic neutralizing activity, colostrum from 2 cows receiving a longer-duration vaccination regimen demonstrated broad HIV-1-neutralizing activity. Colostrum-purified polyclonal IgG retained gp140 reactivity and neutralization activity and blocked the binding of the b12 monoclonal antibody to gp140, showing specificity for the CD4 binding site. Colostrum-derived anti-HIV antibodies offer a cost-effective option for preparing the substantial quantities of broadly neutralizing antibodies that would be needed in a low-cost topical combination HIV-1 microbicide. PMID:22664963

  1. Oxalates as Activating Groups for Alcohols in Visible Light Photoredox Catalysis: Formation of Quaternary Centers by Redox-Neutral Fragment Coupling

    PubMed Central

    MacMillan, David W. C.; Overman, Larry E.

    2015-01-01

    Alkyl oxalates are new bench-stable alcohol-activating groups for radical generation under visible light photoredox conditions. Using these precursors, the first net redox-neutral coupling of tertiary and secondary alcohols with electron-deficient alkenes is achieved. PMID:26322524

  2. Neutral loss analysis of amino acids by desorption electrospray ionization using an unmodified tandem quadrupole mass spectrometer.

    PubMed

    Corso, Gaetano; Paglia, Giuseppe; Garofalo, Daniela; D'Apolito, Oceania

    2007-01-01

    A new method to analyze free amino acids using desorption electrospray ionization (DESI) has been implemented. The method is based on the neutral loss mode determination of underivatized amino acids using a tandem quadrupole mass spectrometer equipped with an unmodified atmospheric interface. Qualitative and quantitative optimization of DESI parameters, including ESI voltage, solvent flow rate, angle of collection and incidence, gas flow and temperatures, was performed for amino acids detection. The parameters for DESI analysis were evaluated using a mixture of valine, leucine, methionine, phenylalanine and tyrosine standards. A few microliters of this mixture were deposited on a slide, dried and analyzed at a flow rate of 2 microL/min. The optimal ionization response was obtained using laboratory glass slides and an equivalent solution of water/methanol doped with 2% of formic acid. The method specificity was evaluated by comparing product ion spectra and neutral loss analysis of amino acids obtained either by DESI or by electrospray ionization flow injection analysis (ESI-FIA). To evaluate the quantitative response on amino acids analyzed by DESI, calibration curves were performed on amino acid standard solutions spiked with a fixed amount of labelled amino acids. The method was also employed to analyze free amino acids from blood spots, after a rapid solvent extraction without other sample pretreatment, from positive and negative subjects. The method enables one to analyze biological samples and to discriminate healthy subjects from patients affected by inherited metabolic diseases. The intrinsic high-throughput analysis of DESI represents an opportunity, because of its potential application in clinical chemistry, for the expanded screening of some inborn errors of metabolism.

  3. Learning the Relationship between the Primary Structure of HIV Envelope Glycoproteins and Neutralization Activity of Particular Antibodies by Using Artificial Neural Networks

    PubMed Central

    Buiu, Cătălin; Putz, Mihai V.; Avram, Speranta

    2016-01-01

    The dependency between the primary structure of HIV envelope glycoproteins (ENV) and the neutralization data for given antibodies is very complicated and depends on a large number of factors, such as the binding affinity of a given antibody for a given ENV protein, and the intrinsic infection kinetics of the viral strain. This paper presents a first approach to learning these dependencies using an artificial feedforward neural network which is trained to learn from experimental data. The results presented here demonstrate that the trained neural network is able to generalize on new viral strains and to predict reliable values of neutralizing activities of given antibodies against HIV-1. PMID:27727189

  4. Structural characterization of neutral oligosaccharides with blood-group A and H activity isolated from bovine submaxillary mucin.

    PubMed Central

    Savage, A V; D'Arcy, S M; Donoghue, C M

    1991-01-01

    In this study we investigated the structures of 11 neutral oligosaccharides released from bovine submaxillary mucin by alkaline borohydride treatment and isolated by h.p.l.c. One hexa-, one penta-, three tetra-, four tri- and two di-saccharides containing core types 1, 2, 3 or 4 were obtained. We report their structures, determined by a combination of one- and two-dimensional 1H n.m.r. spectroscopy at 270 MHz and methylation analysis involving g.l.c.-m.s., along with their approximate molar ratios. Only three of these oligosaccharides have previously been reported in this source. Of the new oligosaccharides, one contains the blood-group-A antigenic determinant, two contain the blood-group-H type 2 determinant, while another contains the blood-group-H type 3 determinant. The oligosaccharide GlcNAc beta (1----6)[GlcNAc beta (1----3)]GalNAcol, although previously found as a core structure, has been isolated here as a novel trisaccharide. PMID:1718265

  5. Frozen translational and rotational motion of human immunodeficiency virus transacting activator of transcription peptide-modified nanocargo on neutral lipid bilayer.

    PubMed

    Wei, Lin; Zhao, Xin; Chen, Bo; Li, Hongchang; Xiao, Lehui; Yeung, Edward S

    2013-05-21

    With time-resolved high-precision single-particle tracking methodologies, we explored the adsorption and thermal motion of transacting activator of transcription (TAT) peptide-modified nanocargo on a model lipid bilayer in the nonelectrostatic domain. We found that the lateral and rotational motion of TAT peptide-modified nanocargo could be effectively suppressed on the surface of neutral lipid membrane, a feature that cannot be explained by existing hypotheses. A semiquantitative association activation energy analysis revealed that multiple weak bonds were required for the initial adsorption process. As a result, the localized multiple TAT peptides on the surface of the nanocargo can provide a pathway for the creation of a net of peptide-lipid complexes (e.g., lipid domain). The dragging forces caused by these complexes effectively confined the thermal motion of the nanocargo on the fluid membrane that cannot be achieved by individual peptides with random spatial and conformational distributions. These interesting findings could provide insightful information for the better understanding of the intracellular internalization mechanism of TAT peptide-modified nanocargo and might shed new light on the development of highly efficient intracellular carriers for site-specific delivery of drugs and genes.

  6. Neutral Atmospheres

    NASA Astrophysics Data System (ADS)

    Mueller-Wodarg, I. C. F.; Strobel, D. F.; Moses, J. I.; Waite, J. H.; Crovisier, J.; Yelle, R. V.; Bougher, S. W.; Roble, R. G.

    This paper summarizes the understanding of aeronomy of neutral atmospheres in the solar system, discussing most planets as well as Saturn's moon Titan and comets. The thermal structure and energy balance is compared, highlighting the principal reasons for discrepancies amongst the atmospheres, a combination of atmospheric composition, heliocentric distance and other external energy sources not common to all. The composition of atmospheres is discussed in terms of vertical structure, chemistry and evolution. The final section compares dynamics in the upper atmospheres of most planets and highlights the importance of vertical dynamical coupling as well as magnetospheric forcing in auroral regions, where present. It is shown that a first order understanding of neutral atmospheres has emerged over the past decades, thanks to the combined effects of spacecraft and Earth-based observations as well as advances in theoretical modeling capabilities. Key gaps in our understanding are highlighted which ultimately call for a more comprehensive programme of observation and laboratory measurements.

  7. Neutral Atmospheres

    NASA Astrophysics Data System (ADS)

    Mueller-Wodarg, I. C. F.; Strobel, D. F.; Moses, J. I.; Waite, J. H.; Crovisier, J.; Yelle, R. V.; Bougher, S. W.; Roble, R. G.

    2008-08-01

    This paper summarizes the understanding of aeronomy of neutral atmospheres in the solar system, discussing most planets as well as Saturn’s moon Titan and comets. The thermal structure and energy balance is compared, highlighting the principal reasons for discrepancies amongst the atmospheres, a combination of atmospheric composition, heliocentric distance and other external energy sources not common to all. The composition of atmospheres is discussed in terms of vertical structure, chemistry and evolution. The final section compares dynamics in the upper atmospheres of most planets and highlights the importance of vertical dynamical coupling as well as magnetospheric forcing in auroral regions, where present. It is shown that a first order understanding of neutral atmospheres has emerged over the past decades, thanks to the combined effects of spacecraft and Earth-based observations as well as advances in theoretical modeling capabilities. Key gaps in our understanding are highlighted which ultimately call for a more comprehensive programme of observation and laboratory measurements.

  8. Activation analysis using Cornell TRIGA

    SciTech Connect

    Hossain, Tim Z.

    1994-07-01

    A major use of the Cornell TRIGA is for activation analysis. Over the years many varieties of samples have been analyzed from a number of fields of interest ranging from geology, archaeology and textiles. More recently the analysis has been extended to high technology materials for applications in optical and semiconductor devices. Trace analysis in high purity materials like Si wafers has been the focus in many instances, while in others analysis of major/minor components were the goals. These analysis has been done using the delayed mode. Results from recent measurements in semiconductors and other materials will be presented. In addition the near future capability of using prompt gamma activation analysis using the Cornell cold neutron beam will be discussed. (author)

  9. Monoclonal Antibody Combinations that Present Synergistic Neutralizing Activity: A Platform for Next-Generation Anti-Toxin Drugs

    PubMed Central

    Diamant, Eran; Torgeman, Amram; Ozeri, Eyal; Zichel, Ran

    2015-01-01

    Monoclonal antibodies (MAbs) are among the fastest-growing therapeutics and are being developed for a broad range of indications, including the neutralization of toxins, bacteria and viruses. Nevertheless, MAbs potency is still relatively low when compared to conventional polyclonal Ab preparations. Moreover, the efficacy of an individual neutralizing MAb may significantly be hampered by the potential absence or modification of its target epitope in a mutant or subtype of the infectious agent. These limitations of individual neutralizing MAbs can be overcome by using oligoclonal combinations of several MAbs with different specificities to the target antigen. Studies conducted in our lab and by others show that such combined MAb preparation may present substantial synergy in its potency over the calculated additive potency of its individual MAb components. Moreover, oligoclonal preparation is expected to be better suited to compensating for reduced efficacy due to epitope variation. In this review, the synergistic neutralization properties of combined oligoclonal Ab preparations are described. The effect of Ab affinity, autologous Fc fraction, and targeting a critical number of epitopes, as well as the unexpected contribution of non-neutralizing clones to the synergistic neutralizing effect are presented and discussed. PMID:26035486

  10. Standardizing Activation Analysis: New Software for Photon Activation Analysis

    SciTech Connect

    Sun, Z. J.; Wells, D.; Green, J.; Segebade, C.

    2011-06-01

    Photon Activation Analysis (PAA) of environmental, archaeological and industrial samples requires extensive data analysis that is susceptible to error. For the purpose of saving time, manpower and minimizing error, a computer program was designed, built and implemented using SQL, Access 2007 and asp.net technology to automate this process. Based on the peak information of the spectrum and assisted by its PAA library, the program automatically identifies elements in the samples and calculates their concentrations and respective uncertainties. The software also could be operated in browser/server mode, which gives the possibility to use it anywhere the internet is accessible. By switching the nuclide library and the related formula behind, the new software can be easily expanded to neutron activation analysis (NAA), charged particle activation analysis (CPAA) or proton-induced X-ray emission (PIXE). Implementation of this would standardize the analysis of nuclear activation data. Results from this software were compared to standard PAA analysis with excellent agreement. With minimum input from the user, the software has proven to be fast, user-friendly and reliable.

  11. Impact of viral attachment factor expression on antibody-mediated neutralization of flaviviruses.

    PubMed

    Obara, Christopher J; Dowd, Kimberly A; Ledgerwood, Julie E; Pierson, Theodore C

    2013-03-01

    Neutralization of flaviviruses requires engagement of the virion by antibodies with a stoichiometry that exceeds a required threshold. Factors that modulate the number of antibodies bound to an individual virion when it contacts target cells impact neutralization potency. However, the contribution of cellular factors to the potency of neutralizing antibodies has not been explored systematically. Here we investigate the relationship between expression level of a viral attachment factor on cells and the neutralizing potency of antibodies. Analysis of the attachment factor DC-SIGNR on cells in neutralization studies failed to identify a correlation between DC-SIGNR expression and antibody-mediated protection. Furthermore, neutralization potency was equivalent on a novel Jurkat cell line induced to express DC-SIGNR at varying levels. Finally, blocking virus-attachment factor interactions had no impact on neutralization activity. Altogether, our studies suggest that cellular attachment factor expression is not a significant contributor to the potency of neutralizing antibodies to flaviviruses.

  12. Modeling the neutral sodium tails of comets

    NASA Astrophysics Data System (ADS)

    Birkett, K.; Jones, G.; Coates, A.

    2014-07-01

    Neutral sodium is typically easy to detect in active comets around perihelion, due to the very high efficiency of the sodium D transition, and, at some comets, a distinct neutral sodium tail is observed. The first distinct neutral sodium tail images were apparent in C/1995 O1 (Hale-Bopp) data taken using CoCam [1], but, since this initial detection, similar features have been observed at near-Sun comets using the LASCO coronagraph on SOHO. A full picture of the distribution and evolution of neutral cometary sodium may best be established using a combination of spectra and images in different filters at multiple times throughout the orbit. The high efficiency of the sodium D transition has allowed it to be detected in systems, even if the column density of sodium is extremely low. In these instances it is sometimes possible to determine some of the system's characteristics from the sodium emission detection, such as in Io's plasma torus [2] and Enceladus's plume [3,4]. It is hoped that a similar approach may be applied to the active cometary environment, but, at present, the production of neutral sodium is unknown. Various authors [5--9, thorough review presented in 10] have suggested various combinations of sources of neutral sodium in the nuclear region, near-nuclear region, dust tail, and ion tail. The morphology and evolution of the neutral cometary sodium tail are difficult to intuitively predict due to the Swings and Greenstein effects. In order to understand the wide variety of cometary observations of neutral sodium available we have developed the first fully three-dimensional, heliocentric-distance-dependent, versatile Monte Carlo neutral sodium tail model, which incorporates the unintuitive variation in radiation pressure influences on sodium atoms with different heliocentric velocities. Our model was initially based on that of Brown et al [7]. We present preliminary results from this model. We have found initial agreement with the overall morphology and

  13. Antibodies with specificity to native gp120 and neutralization activity against primary human immunodeficiency virus type 1 isolates elicited by immunization with oligomeric gp160.

    PubMed Central

    VanCott, T C; Mascola, J R; Kaminski, R W; Kalyanaraman, V; Hallberg, P L; Burnett, P R; Ulrich, J T; Rechtman, D J; Birx, D L

    1997-01-01

    Current human immunodeficiency virus type 1 (HIV-1) envelope vaccine candidates elicit high antibody binding titers with neutralizing activity against T-cell line-adapted but not primary HIV-1 isolates. Serum antibodies from these human vaccine recipients were also found to be preferentially directed to linear epitopes within gp120 that are poorly exposed on native gp120. Systemic immunization of rabbits with an affinity-purified oligomeric gp160 protein formulated with either Alhydrogel or monophosphoryl lipid A-containing adjuvants resulted in the induction of high-titered serum antibodies that preferentially bound epitopes exposed on native forms of gp120 and gp160, recognized a restricted number of linear epitopes, efficiently bound heterologous strains of monomeric gp120 and cell surface-expressed oligomeric gp120/gp41, and neutralized several strains of T-cell line-adapted HIV-1. Additionally, those immune sera with the highest oligomeric gp160 antibody binding titers had neutralizing activity against some primary HIV-1 isolates, using phytohemagglutinin-stimulated peripheral blood mononuclear cell targets. Induction of an antibody response preferentially reactive with natively folded gp120/gp160 was dependent on the tertiary structure of the HIV-1 envelope immunogen as well as its adjuvant formulation, route of administration, and number of immunizations administered. These studies demonstrate the capacity of a soluble HIV-1 envelope glycoprotein vaccine to elicit an antibody response capable of neutralizing primary HIV-1 isolates. PMID:9151820

  14. Betaglycan has two independent domains required for high affinity TGF-β binding: proteolytic cleavage separates the domains and inactivates the neutralizing activity of the soluble receptor

    PubMed Central

    Mendoza, Valentín; Vilchis-Landeros, M. Magdalena; Mendoza-Hernández, Guillermo; Huang, Tao; Villarreal, Maria M.; Hinck, Andrew P.; López-Casillas, Fernando; Montiel, Jose-Luis

    2009-01-01

    Summary Betaglycan is a co-receptor for members of the TGF-β superfamily. Mutagenesis has identified two ligand binding regions, one at the membrane-distal and the other at the membrane-proximal half of the betaglycan ectodomain. Here we show that partial plasmin digestion of soluble betaglycan produces two proteolysis-resistant fragments of 45 and 55 kDa, consistent with the predicted secondary structure, which indicates an intervening non-structured linker region separating the highly structured N- and C-terminal domains. Amino terminal sequencing indicates that the 45 and 55 kDa fragments correspond, respectively, to the membrane-distal and -proximal regions. Plasmin treatment of membrane betaglycan results in the production of equivalent proteolysis-resistant fragments. The 45 and 55 kDa fragments, as well as their recombinant soluble counterparts, Sol Δ10 and Sol Δ11, bind TGF-β, nonetheless, compared to intact soluble betaglycan, have severely diminished ability to block TGF-β activity. Surface plasmon resonance (SPR) analysis indicates that soluble betaglycan has Kds in the low nanomolar range for the three TGF-β isoforms, while those for Sol Δ10 and Sol Δ11 are 1 – 2 orders of magnitude higher. SPR analysis further shows that the Kds of Sol Δ11 are not changed in the presence of Sol Δ10, indicating that the high affinity of soluble betaglycan is a consequence of tethering of the domains together. Overall, these results, suggest that betaglycan ectodomain exhibits a bi-lobular structure in which each lobule folds independently, binds TGF-β through distinct non-overlapping interfaces, and that linker modification may be an approach to improve soluble betaglycan’s TGF-β neutralizing activity. PMID:19842711

  15. Augmentation of the Lipopolysaccharide-Neutralizing Activities of Human Cathelicidin CAP18/LL-37-Derived Antimicrobial Peptides by Replacement with Hydrophobic and Cationic Amino Acid Residues

    PubMed Central

    Nagaoka, Isao; Hirota, Satoko; Niyonsaba, François; Hirata, Michimasa; Adachi, Yoshiyuki; Tamura, Hiroshi; Tanaka, Shigenori; Heumann, Didier

    2002-01-01

    Mammalian myeloid and epithelial cells express various peptide antibiotics (such as defensins and cathelicidins) that contribute to the innate host defense against invading microorganisms. Among these peptides, human cathelicidin CAP18/LL-37 (L1 to S37) possesses not only potent antibacterial activity against gram-positive and gram-negative bacteria but also the ability to bind to gram-negative lipopolysaccharide (LPS) and neutralize its biological activities. In this study, to develop peptide derivatives with improved LPS-neutralizing activities, we utilized an 18-mer peptide (K15 to V32) of LL-37 as a template and evaluated the activities of modified peptides by using the CD14+ murine macrophage cell line RAW 264.7 and the murine endotoxin shock model. By replacement of E16 and K25 with two L residues, the hydrophobicity of the peptide (18-mer LL) was increased, and by further replacement of Q22, D26, and N30 with three K residues, the cationicity of the peptide (18-mer LLKKK) was enhanced. Among peptide derivatives, 18-mer LLKKK displayed the most powerful LPS-neutralizing activity: it was most potent at binding to LPS, inhibiting the interaction between LPS and LPS-binding protein, and attaching to the CD14 molecule, thereby suppressing the binding of LPS to CD14+ cells and attenuating production of tumor necrosis factor alpha (TNF-α) by these cells. Furthermore, in the murine endotoxin shock model, 18-mer LLKKK most effectively suppressed LPS-induced TNF-α production and protected mice from lethal endotoxin shock. Together, these observations indicate that the LPS-neutralizing activities of the amphipathic human CAP18/LL-37-derived 18-mer peptide can be augmented by modifying its hydrophobicity and cationicity, and that 18-mer LLKKK is the most potent of the peptide derivatives, with therapeutic potential for gram-negative bacterial endotoxin shock. PMID:12204946

  16. Search for active neutrino disappearance using neutral-current interactions in the MINOS long-baseline experiment.

    PubMed

    Adamson, P; Andreopoulos, C; Arms, K E; Armstrong, R; Auty, D J; Ayres, D S; Backhouse, C; Baller, B; Barr, G; Barrett, W L; Becker, B R; Belias, A; Bernstein, R H; Bhattacharya, D; Bishai, M; Blake, A; Bock, G J; Boehm, J; Boehnlein, D J; Bogert, D; Bower, C; Buckley-Geer, E; Cavanaugh, S; Chapman, J D; Cherdack, D; Childress, S; Choudhary, B C; Cobb, J H; Coleman, S J; Culling, A J; de Jong, J K; Dierckxsens, M; Diwan, M V; Dorman, M; Dytman, S A; Escobar, C O; Evans, J J; Harris, E Falk; Feldman, G J; Frohne, M V; Gallagher, H R; Godley, A; Goodman, M C; Gouffon, P; Gran, R; Grashorn, E W; Grossman, N; Grzelak, K; Habig, A; Harris, D; Harris, P G; Hartnell, J; Hatcher, R; Heller, K; Himmel, A; Holin, A; Hsu, L; Hylen, J; Irwin, G M; Ishitsuka, M; Jaffe, D E; James, C; Jensen, D; Kafka, T; Kasahara, S M S; Kim, J J; Kim, M S; Koizumi, G; Kopp, S; Kordosky, M; Koskinen, D J; Kotelnikov, S K; Kreymer, A; Kumaratunga, S; Lang, K; Ling, J; Litchfield, P J; Litchfield, R P; Loiacono, L; Lucas, P; Ma, J; Mann, W A; Marchionni, A; Marshak, M L; Marshall, J S; Mayer, N; McGowan, A M; Meier, J R; Messier, M D; Metelko, C J; Michael, D G; Miller, W H; Mishra, S R; Moore, C D; Morfín, J; Mualem, L; Mufson, S; Murgia, S; Musser, J; Naples, D; Nelson, J K; Newman, H B; Nichol, R J; Nicholls, T C; Ochoa-Ricoux, J P; Oliver, W P; Ospanov, R; Paley, J; Paolone, V; Para, A; Patzak, T; Pavlović, Z; Pawloski, G; Pearce, G F; Peck, C W; Petyt, D A; Pittam, R; Plunkett, R K; Rahaman, A; Rameika, R A; Raufer, T M; Rebel, B; Reichenbacher, J; Rodrigues, P A; Rosenfeld, C; Rubin, H A; Ryabov, V A; Sanchez, M C; Saoulidou, N; Schneps, J; Schreiner, P; Shanahan, P; Smart, W; Smith, C; Sousa, A; Speakman, B; Stamoulis, P; Strait, M; Tagg, N; Talaga, R L; Tavera, M A; Thomas, J; Thomson, M A; Thron, J L; Tinti, G; Trostin, I; Tsarev, V A; Tzanakos, G; Urheim, J; Vahle, P; Viren, B; Ward, D R; Watabe, M; Weber, A; Webb, R C; Wehmann, A; West, N; White, C; Wojcicki, S G; Wright, D M; Yang, T; Zhang, K; Zwaska, R

    2008-11-28

    We report the first detailed comparisons of the rates and spectra of neutral-current neutrino interactions at two widely separated locations. A depletion in the rate at the far site would indicate mixing between nu(mu) and a sterile particle. No anomalous depletion in the reconstructed energy spectrum is observed. Assuming oscillations occur at a single mass-squared splitting, a fit to the neutral- and charged-current energy spectra limits the fraction of nu(mu) oscillating to a sterile neutrino to be below 0.68 at 90% confidence level. A less stringent limit due to a possible contribution to the measured neutral-current event rate at the far site from nu(e) appearance at the current experimental limit is also presented.

  17. Crystal Structure of PG16 and Chimeric Dissection with Somatically Related PG9: Structure-Function Analysis of Two Quaternary-Specific Antibodies That Effectively Neutralize HIV-1

    SciTech Connect

    Pancera, Marie; McLellan, Jason S.; Wu, Xueling; Zhu, Jiang; Changela, Anita; Schmidt, Stephen D.; Yang, Yongping; Zhou, Tongqing; Phogat, Sanjay; Mascola, John R.; Kwong, Peter D.

    2010-11-03

    HIV-1 resists neutralization by most antibodies. Two somatically related human antibodies, PG9 and PG16, however, each neutralize 70 to 80% of circulating HIV-1 isolates. Here we present the structure of the antigen-binding fragment of PG16 in monoclinic and orthorhombic lattices at 2.4 and 4.0 {angstrom}, respectively, and use a combination of structural analysis, paratope dissection, and neutralization assessment to determine the functional relevance of three unusual PG9/PG16 features: N-linked glycosylation, extensive affinity maturation, and a heavy chain-third complementarity-determining region (CDR H3) that is one of the longest observed in human antibodies. Glycosylation extended off the side of the light chain variable domain and was not required for neutralization. The CDR H3 formed an axe-shaped subdomain, which comprised 42% of the CDR surface, with the axe head looming {approx}20 {angstrom} above the other combining loops. Comprehensive sets of chimeric swaps between PG9 and PG16 of light chain, heavy chain, and CDR H3 were employed to decipher structure-function relationships. Chimeric swaps generally complemented functionally, with differences in PG9/PG16 neutralization related primarily to residue differences in CDR H3. Meanwhile, chimeric reversions to genomic V genes showed isolate-dependent effects, with affinity maturation playing a significant role in augmenting neutralization breadth (P = 0.036) and potency (P < 0.0001). The structural and functional details of extraordinary CDR H3 and extensive affinity maturation provide insights into the neutralization mechanism of and the elicitation pathway for broadly neutralizing antibodies like PG9 and PG16.

  18. Appreciation of Hardy's Poem "Neutral Tones" from the Point of View of Functional Analysis

    ERIC Educational Resources Information Center

    Chen, Hong-qin

    2008-01-01

    According to the theory of Systemic-Functional Linguistics (SFL), language cannot be disassociated from meaning. Function and semantics, as SFL suggests, are considered as the basis of human language and communicative activity. In order to reveal the inseparability of language and semantic, this paper aims to analyze and appreciate Hardy's poem…

  19. Gender Neutralities, Dichotomies and Hidden Inequalities: Analysis of Vocational Teachers' Reflections on Gender in the Profession

    ERIC Educational Resources Information Center

    Lappalainen, Sirpa; Lahelma, Elina; Pehkonen, Leila; Isopahkala-Bouret, Ulpukka

    2012-01-01

    This article analyses how Finnish vocational teachers make sense of the meanings of gender in their work. The context of the study consists of the two most gender segregated environments of vocational education: the female-dominated Sector of Health and Social Services and the male-dominated Sector of Technology and Transport. Our analysis draws…

  20. Correcting the Record on the Analysis of IBEX and STEREO Data Regarding Variations in the Neutral Interstellar Wind

    NASA Astrophysics Data System (ADS)

    Frisch, P. C.; Bzowski, M.; Drews, C.; Leonard, T.; Livadiotis, G.; McComas, D. J.; Möbius, E.; Schwadron, N.; Sokół, J. M.

    2015-03-01

    The journey of the Sun through space carries the solar system through a dynamic interstellar environment that is presently characterized by a Mach ~1 motion between the heliosphere and the surrounding warm partially ionized interstellar cloud. The interaction between the heliosphere and interstellar medium is an evolving process due to variable solar wind properties and the turbulent nature of the interstellar cloud that surrounds the heliosphere. Frisch et al. presented a meta-analysis of the historical data on the interstellar wind flowing through the heliosphere and concluded that temporal changes in the ecliptic longitude of the flow direction with time were statistically indicated by the data available in the refereed literature at the time of that writing. Lallement & Bertaux disagree with this result, and suggested, for instance, that a key instrumental response function of IBEX-Lo was incorrect and that the STEREO pickup ion data are unsuitable for diagnosing the flow of interstellar neutrals through the heliosphere. In this paper we first show that temporal variations in the interstellar wind through the heliosphere are consistent with our knowledge of the very local interstellar medium. The statistical analysis of the helium wind data is revisited, and a recent correction of a typographical error in the literature is incorporated into the new fits. With this correction, and including no newer IBEX results, these combined data still indicate that a change in the longitude of the interstellar neutral wind of λ = 5.°6 ± 2.°4 over the past forty years remains statistically likely, but an constant flow longitude is now statistically possible. Other scenarios for the selection of subsets of these data used in the fitting process produce similar conclusions. We show that the speculations made by Lallement & Bertin about the IBEX instrumental response function are incorrect, and that their other objections to the data used in the meta-analysis are either

  1. Neutralization activity and kinetics of two broad-range human monoclonal IgG1 derived from recombinant Fab fragments and directed against Hepatitis C virus E2 glycoprotein.

    PubMed

    Diotti, Roberta Antonia; Sautto, Giuseppe Andrea; Solforosi, Laura; Mancini, Nicasio; Clementi, Massimo; Burioni, Roberto

    2012-10-01

    Hepatitis C virus (HCV) is the major cause of chronic liver disease worldwide. There is evidence that neutralizing anti-HCV antibodies may find potential applications in novel prophylactic and therapeutic strategies. This paper describes the very high neutralization activity and unique biological features of two broadly cross-reactive and cross-neutralizing anti-HCV human monoclonal IgG1 derived from human monoclonal recombinant Fab fragments.

  2. Kinetic analysis about the effects of neutral salts on the thermal stability of yeast alcohol dehydrogenase.

    PubMed

    Ikegaya, Kazuo

    2005-03-01

    The effects of salts on the rate constants of inactivation by heat of yeast alcohol dehydrogenase (YADH) at 60.0 degrees C were measured. Different effects were observed at low and high salt concentrations. At high concentrations, some salts had stabilizing effects, while others were destabilizing. The effects of salts in the high concentration range examined can be described as follows: (decreased thermal stability) NaClO(4) < NaI = (C(2)H(5))(4)NBr < NH(4)Br < NaBr = KBr = CsBr = (no addition) < (CH(3))(4)NBr < KCl < KF < Na(2)SO(4) (increased thermal stability). The decreasing effect of NaClO(4) on YADH controlled the thermal stability of the enzyme absolutely and was not compensated by the addition of Na(2)SO(4), a salt which stabilized the enzyme. However, Na(2)SO(4) compensation did occur in response to the decrease in thermal stability caused by (C(2)H(5))(4)NBr. The rate constants of inactivation by heat (k (in)) of the enzyme were measured at various temperatures. Effective values of the thermodynamic activation parameters of thermal inactivation, activation of free energy (DeltaG (double dagger)), activation enthalpy (DeltaH (double dagger)), and activation entropy (DeltaS (double dagger)), were determined. The thermal stability of YADH in 0.8 M Na(2)SO(4) increased more than that of pyruvate kinase from Bacillus stearothermophilus, a moderate thermophile. The changes in the values of DeltaH (double dagger) and DeltaS (double dagger) were great and showed a general compensatory tendency, with the exception of in the case of NaClO(4). The temperature for the general compensation effect (T (c)) was approximately 123 degrees C. With Na(2)SO(4), the thermal stability of YADH at a temperature below T (c) was greater than that in the absence of salt due to the higher values of DeltaH (double dagger) and DeltaS (double dagger), respectively, and thus was an example of low-temperature enzymatic stabilization. With (C(2)H(5))(4)NBr, the thermal stability of YADH

  3. Neutralized nanoparticle composed of SS-cleavable and pH-activated lipid-like material as a long-lasting and liver-specific gene delivery system.

    PubMed

    Ukawa, Masami; Akita, Hidetaka; Hayashi, Yasuhiro; Ishiba, Ryohei; Tange, Kota; Arai, Masaya; Kubo, Kazuhiro; Higuchi, Yuriko; Shimizu, Kazunori; Konishi, Satoshi; Hashida, Mitsuru; Harashima, Hideyoshi

    2014-08-01

    Charge-neutralized lipid envelope-type nanoparticles formed with SS-cleavable and pH-activated lipid-like materials (ssPalm) accumulate rapidly in the liver without forming aggregates in the blood circulation, and result in a liver-specific gene expression for a long duration (>2 weeks) with neither immunological responses nor hepatotoxicity after intraveneous administration, when it carries pDNA free from CpG-motifs.

  4. ACDOS1: a computer code to calculate dose rates from neutron activation of neutral beamlines and other fusion-reactor components

    SciTech Connect

    Keney, G.S.

    1981-08-01

    A computer code has been written to calculate neutron induced activation of neutral-beam injector components and the corresponding dose rates as a function of geometry, component composition, and time after shutdown. The code, ACDOS1, was written in FORTRAN IV to calculate both activity and dose rates for up to 30 target nuclides and 50 neutron groups. Sufficient versatility has also been incorporated into the code to make it applicable to a variety of general activation problems due to neutrons of energy less than 20 MeV.

  5. Numerical analysis of the angular motion of a neutrally buoyant spheroid in shear flow at small Reynolds numbers.

    PubMed

    Rosén, T; Einarsson, J; Nordmark, A; Aidun, C K; Lundell, F; Mehlig, B

    2015-12-01

    We numerically analyze the rotation of a neutrally buoyant spheroid in a shear flow at small shear Reynolds number. Using direct numerical stability analysis of the coupled nonlinear particle-flow problem, we compute the linear stability of the log-rolling orbit at small shear Reynolds number Re(a). As Re(a)→0 and as the box size of the system tends to infinity, we find good agreement between the numerical results and earlier analytical predictions valid to linear order in Re(a) for the case of an unbounded shear. The numerical stability analysis indicates that there are substantial finite-size corrections to the analytical results obtained for the unbounded system. We also compare the analytical results to results of lattice Boltzmann simulations to analyze the stability of the tumbling orbit at shear Reynolds numbers of order unity. Theory for an unbounded system at infinitesimal shear Reynolds number predicts a bifurcation of the tumbling orbit at aspect ratio λ(c)≈0.137 below which tumbling is stable (as well as log rolling). The simulation results show a bifurcation line in the λ-Re(a) plane that reaches λ≈0.1275 at the smallest shear Reynolds number (Re(a)=1) at which we could simulate with the lattice Boltzmann code, in qualitative agreement with the analytical results. PMID:26764819

  6. Numerical analysis of the angular motion of a neutrally buoyant spheroid in shear flow at small Reynolds numbers

    NASA Astrophysics Data System (ADS)

    Rosén, T.; Einarsson, J.; Nordmark, A.; Aidun, C. K.; Lundell, F.; Mehlig, B.

    2015-12-01

    We numerically analyze the rotation of a neutrally buoyant spheroid in a shear flow at small shear Reynolds number. Using direct numerical stability analysis of the coupled nonlinear particle-flow problem, we compute the linear stability of the log-rolling orbit at small shear Reynolds number Rea. As Rea→0 and as the box size of the system tends to infinity, we find good agreement between the numerical results and earlier analytical predictions valid to linear order in Rea for the case of an unbounded shear. The numerical stability analysis indicates that there are substantial finite-size corrections to the analytical results obtained for the unbounded system. We also compare the analytical results to results of lattice Boltzmann simulations to analyze the stability of the tumbling orbit at shear Reynolds numbers of order unity. Theory for an unbounded system at infinitesimal shear Reynolds number predicts a bifurcation of the tumbling orbit at aspect ratio λc≈0.137 below which tumbling is stable (as well as log rolling). The simulation results show a bifurcation line in the λ -Rea plane that reaches λ ≈0.1275 at the smallest shear Reynolds number (Rea=1 ) at which we could simulate with the lattice Boltzmann code, in qualitative agreement with the analytical results.

  7. Quetiapine Inhibits Microglial Activation by Neutralizing Abnormal STIM1-Mediated Intercellular Calcium Homeostasis and Promotes Myelin Repair in a Cuprizone-Induced Mouse Model of Demyelination

    PubMed Central

    Wang, Hanzhi; Liu, Shubao; Tian, Yanping; Wu, Xiyan; He, Yangtao; Li, Chengren; Namaka, Michael; Kong, Jiming; Li, Hongli; Xiao, Lan

    2015-01-01

    Microglial activation has been considered as a crucial process in the pathogenesis of neuroinflammation and psychiatric disorders. Several antipsychotic drugs (APDs) have been shown to display inhibitory effects on microglial activation in vitro, possibly through the suppression of elevated intracellular calcium (Ca2+) concentration. However, the exact underlying mechanisms still remain elusive. In this study, we aimed to investigate the inhibitory effects of quetiapine (Que), an atypical APD, on microglial activation. We utilized a chronic cuprizone (CPZ)-induced demyelination mouse model to determine the direct effect of Que on microglial activation. Our results showed that treatment with Que significantly reduced recruitment and activation of microglia/macrophage in the lesion of corpus callosum and promoted remyelination after CPZ withdrawal. Our in vitro studies also confirmed the direct effect of Que on lipopolysaccharide (LPS)-induced activation of microglial N9 cells, whereby Que significantly inhibited the release of nitric oxide (NO) and tumor necrosis factor α (TNF-α). Moreover, we demonstrated that pretreatment with Que, neutralized the up-regulation of STIM1 induced by LPS and declined both LPS and thapsigargin (Tg)-induced store-operated Ca2+ entry (SOCE). Finally, we found that pretreatment with Que significantly reduced the translocation of nuclear factor kappa B (NF-κB) p65 subunit from cytoplasm to nuclei in LPS-activated primary microglial cells. Overall, our data suggested that Que may inhibit microglial activation by neutralization of the LPS-induced abnormal STIM1-mediated intercellular calcium homeostasis. PMID:26732345

  8. Surface analysis of rubbers and plastics using secondary neutral mass spectrometry.

    SciTech Connect

    Pellin, M. J.; Savina, M. R.

    1999-08-23

    Rubber and plastics are complex mixtures of long-chain polymers, smaller organic additives used as plasticizers, mildicides, fungicides, colorants, etc., and inorganic additives such as carbon and silica. Surface analysis of such materials is at once difficult and important. The difficulty lies in the need to identify specific carbon molecules on a very similar organic surface. The importance arises from the dramatic effect that the distribution and concentration of additives have on the physical properties of polymeric materials. Recently we have had some success in applying laser desorption post-ionization mass spectrometry in measuring the surface concentration and distribution of both additives and the polymer molecules themselves. The key has been to use the photoionization properties of the analyte of interest to augment mass spectrometric information.

  9. Use of a Synthetic Biosensor for Neutralizing Activity-Biased Selection of Monoclonal Antibodies against Atroxlysin-I, an Hemorrhagic Metalloproteinase from Bothrops atrox Snake Venom

    PubMed Central

    Schneider, Francisco Santos; Nguyen, Dung Le; Castro, Karen Larissa; Cobo, Sandra; Machado de Avila, Ricardo Andrez; Ferreira, Nivia de Assis; Sanchez, Eladio Flores; Nguyen, Christophe; Granier, Claude; Galéa, Pascale; Chávez-Olortegui, Carlos; Molina, Franck

    2014-01-01

    Background The snake Bothrops atrox is responsible for the majority of envenomings in the northern region of South America. Severe local effects, including hemorrhage, which are mainly caused by snake venom metalloproteinases (SVMPs), are not fully neutralized by conventional serum therapy. Little is known about the immunochemistry of the P-I SVMPs since few monoclonal antibodies (mAbs) against these molecules have been obtained. In addition, producing toxin-neutralizing mAbs remains very challenging. Methodology/Principal Findings Here, we report on the set-up of a functional screening based on a synthetic peptide used as a biosensor to select neutralizing mAbs against SVMPs and the successful production of neutralizing mAbs against Atroxlysin-I (Atr-I), a P-I SVMP from B. atrox. Hybridomas producing supernatants with inhibitory effect against the proteolytic activity of Atr-I towards the FRET peptide Abz-LVEALYQ-EDDnp were selected. Six IgG1 Mabs were obtained (named mAbatr1 to mAbatr6) and also two IgM. mAbatrs1, 2, 3 and 6 were purified. All showed a high specific reactivity, recognizing only Atr-I and B. atrox venom in ELISA and a high affinity, showing equilibrium constants in the nM range for Atr-I. These mAbatrs were not able to bind to Atr-I overlapping peptides, suggesting that they recognize conformational epitopes. Conclusions/Significance For the first time a functional screening based on a synthetic biosensor was successfully used for the selection of neutralizing mAbs against SVMPs. PMID:24762927

  10. Comparison of plaque- and enzyme-linked immunospot-based assays to measure the neutralizing activities of monoclonal antibodies specific to domain III of dengue virus envelope protein.

    PubMed

    Liu, Lidong; Wen, Kun; Li, Jie; Hu, Dongmei; Huang, Yanfen; Qiu, Liwen; Cai, Jianpiao; Che, Xiaoyan

    2012-01-01

    The plaque reduction neutralization test (PRNT) is used widely to measure the neutralization activity of anti-dengue virus (DENV) antibodies, but it is time-consuming and labor-intensive and has low sample throughput. For fast and convenient measurement of neutralizing antibodies, especially in evaluating the efficiency of the DENV vaccines on a large scale, a new method is needed to replace PRNT. In recent decades, several microneutralization assays have been developed to overcome the limitations of PRNT. In the present study, we evaluated one of these, the enzyme-linked immunospot microneutralization test (ELISPOT-MNT), in comparison with PRNT. ELISPOT-MNT is performed in 96-well format, and the plaques are developed after 2 to 4 days using an ELISA to transform them into spots, which are detected automatically with an ELISPOT instrument. The assay is faster than PRNT, has a high throughput, and is more objective. We used 10 monoclonal antibodies (MAbs) against domain III of the DENV envelope protein (EDIII) to evaluate the two assays; all of these MAbs cross-react with all four serotypes of DENV as measured by immunofluorescence assay. The two neutralization assays were performed simultaneously to measure the 50% inhibitory concentration (IC(50)) of these MAbs. Using PRNT as the reference and treating IC(50) values higher than 50 μg/ml of MAbs as negative, ELISPOT-MNT showed a sensitivity of 95.6% and specificity of 88.24% when 10 MAbs were tested against four DENV serotype strains. A good correlation (R(2) = 0.672; P = 0.000) was observed between the two assays, making ELISPOT-MNT a potentially valuable method for measure of neutralizing antibodies against DENV.

  11. Comparison of Plaque- and Enzyme-Linked Immunospot-Based Assays To Measure the Neutralizing Activities of Monoclonal Antibodies Specific to Domain III of Dengue Virus Envelope Protein

    PubMed Central

    Liu, Lidong; Wen, Kun; Li, Jie; Hu, Dongmei; Huang, Yanfen; Qiu, Liwen; Cai, Jianpiao

    2012-01-01

    The plaque reduction neutralization test (PRNT) is used widely to measure the neutralization activity of anti-dengue virus (DENV) antibodies, but it is time-consuming and labor-intensive and has low sample throughput. For fast and convenient measurement of neutralizing antibodies, especially in evaluating the efficiency of the DENV vaccines on a large scale, a new method is needed to replace PRNT. In recent decades, several microneutralization assays have been developed to overcome the limitations of PRNT. In the present study, we evaluated one of these, the enzyme-linked immunospot microneutralization test (ELISPOT-MNT), in comparison with PRNT. ELISPOT-MNT is performed in 96-well format, and the plaques are developed after 2 to 4 days using an ELISA to transform them into spots, which are detected automatically with an ELISPOT instrument. The assay is faster than PRNT, has a high throughput, and is more objective. We used 10 monoclonal antibodies (MAbs) against domain III of the DENV envelope protein (EDIII) to evaluate the two assays; all of these MAbs cross-react with all four serotypes of DENV as measured by immunofluorescence assay. The two neutralization assays were performed simultaneously to measure the 50% inhibitory concentration (IC50) of these MAbs. Using PRNT as the reference and treating IC50 values higher than 50 μg/ml of MAbs as negative, ELISPOT-MNT showed a sensitivity of 95.6% and specificity of 88.24% when 10 MAbs were tested against four DENV serotype strains. A good correlation (R2 = 0.672; P = 0.000) was observed between the two assays, making ELISPOT-MNT a potentially valuable method for measure of neutralizing antibodies against DENV. PMID:22116689

  12. CTEPP STANDARD OPERATING PROCEDURE FOR EXTRACTING AND PREPARING AIR SAMPLES FOR ANALYSIS OF NEUTRAL PERSISTENT ORGANIC POLLUTANTS (SOP-5.12)

    EPA Science Inventory

    The method is for extracting an indoor and outdoor air sample consisting of a quartz fiber filter and an XAD-2 cartridge for analysis of neutral persistent organic pollutants. It covers the extraction and concentration of samples that are to be analyzed by gas chromatography/mass...

  13. CTEPP STANDARD OPERATING PROCEDURE FOR EXTRACTING AND PREPARING DERMAL WIPE SAMPLES FOR ANALYSIS OF NEUTRAL PERSISTENT ORGANIC POLLUTANTS (SOP-5.16)

    EPA Science Inventory

    The method for extracting and preparing a dermal (hand) wipe sample for analysis of neutral persistent organic pollutants is summarized in this SOP. It covers the extraction and concentration of samples that are to be analyzed by gas chromatography/mass spectrometry.

  14. CTEPP STANDARD OPERATING PROCEDURE FOR EXTRACTING AND PREPARING SURFACE WIPE FOR ANALYSIS OF NEUTRAL PERSISTENT ORGANIC POLLUTANTS (SOP-5.17)

    EPA Science Inventory

    This SOP summarizes the method for extracting and preparing a hard floor surface wipe or food preparation surface wipe sample for analysis of neutral persistent organic. It covers the extraction and concentration of samples that are to be analyzed by gas chromatography/mass spect...

  15. Neutron Activation Analysis: Techniques and Applications

    NASA Astrophysics Data System (ADS)

    MacLellan, Ryan

    2011-04-01

    The role of neutron activation analysis in low-energy low-background experimentsis discussed in terms of comparible methods. Radiochemical neutron activation analysis is introduce. The procedure of instrumental neutron activation analysis is detailed especially with respect to the measurement of trace amounts of natural radioactivity. The determination of reactor neutron spectrum parameters required for neutron activation analysis is also presented.

  16. Neutron Activation Analysis: Techniques and Applications

    SciTech Connect

    MacLellan, Ryan

    2011-04-27

    The role of neutron activation analysis in low-energy low-background experimentsis discussed in terms of comparible methods. Radiochemical neutron activation analysis is introduce. The procedure of instrumental neutron activation analysis is detailed especially with respect to the measurement of trace amounts of natural radioactivity. The determination of reactor neutron spectrum parameters required for neutron activation analysis is also presented.

  17. Constraining the Europa Neutral Torus

    NASA Astrophysics Data System (ADS)

    Smith, Howard T.; Mitchell, Donald; mauk, Barry; Johnson, Robert E.; clark, george

    2016-10-01

    "Neutral tori" consist of neutral particles that usually co-orbit along with their source forming a toroidal (or partial toroidal) feature around the planet. The distribution and composition of these features can often provide important, if not unique, insight into magnetospheric particles sources, mechanisms and dynamics. However, these features can often be difficult to directly detect. One innovative method for detecting neutral tori is by observing Energetic Neutral Atoms (ENAs) that are generally considered produced as a result of charge exchange interactions between charged and neutral particles.Mauk et al. (2003) reported the detection of a Europa neutral particle torus using ENA observations. The presence of a Europa torus has extremely large implications for upcoming missions to Jupiter as well as understanding possible activity at this moon and providing critical insight into what lies beneath the surface of this icy ocean world. However, ENAs can also be produced as a result of charge exchange interactions between two ionized particles and in that case cannot be used to infer the presence of neutral particle population. Thus, a detailed examination of all possible source interactions must be considered before one can confirm that likely original source population of these ENA images is actually a Europa neutral particle torus. For this talk, we examine the viability that the Mauk et al. (2003) observations were actually generated from a neutral torus emanating from Europa as opposed to charge particle interactions with plasma originating from Io. These results help constrain such a torus as well as Europa source processes.

  18. Structural Analysis of Human and Macaque Monoclonal Antibodies 2909 and 2.5B: Implications for the Configuration of the Quaternary Neutralizing Epitope of HIV-1 gp120

    SciTech Connect

    B Spurrier; J Sampson; M Totrov; H Li; T ONeal; C Williams; J Robinson; M Gorny; S Zolla-Pazner; X Kong

    2011-12-31

    The quaternary neutralizing epitope (QNE) of HIV-1 gp120 is preferentially expressed on the trimeric envelope spikes of intact HIV virions, and QNE-specific monoclonal antibodies (mAbs) potently neutralize HIV-1. Here, we present the crystal structures of the Fabs of human mAb 2909 and macaque mAb 2.5B. Both mAbs have long beta hairpin CDR H3 regions >20 {angstrom} in length that are each situated at the center of their respective antigen-binding sites. Computational analysis showed that the paratopes include the whole CDR H3, while additional CDR residues form shallow binding pockets. Structural modeling suggests a way to understand the configuration of QNEs and the antigen-antibody interaction for QNE mAbs. Our data will be useful in designing immunogens that may elicit potent neutralizing QNE Abs.

  19. Structural Analysis of Human and Macaque Monoclonal Antibodies 2909 and 2.5B: Implications for the Configuration of the Quaternary Neutralizing Epitope of HIV-1 gp120

    PubMed Central

    Spurrier, Brett; Sampson, Jared M.; Totrov, Maxim; Li, Huiguang; O'Neal, Timothy; Williams, Constance; Robinson, James; Gorny, Miroslaw K.; Zolla-Pazner, Susan; Kong, Xiang-Peng

    2011-01-01

    Summary The quaternary neutralizing epitope (QNE) of HIV-1 gp120 is preferentially expressed on the trimeric envelope spikes of intact HIV virions, and QNE-specific monoclonal antibodies (mAbs) potently neutralize HIV-1. Here we present the crystal structures of the Fabs of human mAb 2909 and macaque mAb 2.5B. Both mAbs have long beta hairpin CDR H3 regions >20Å in length that are each situated at the center of their respective antigen-binding sites. Computational analysis showed that the paratopes include the whole CDR H3, while additional CDR residues form shallow binding pockets. Structural modeling suggests a way to understand the configuration of QNEs and the antigen antibody interaction for QNE mAbs. Our data will be useful in designing immunogens that may elicit potent neutralizing QNE Abs. PMID:21565703

  20. New Organocatalyst Scaffolds with High Activity in Promoting Hydrazone and Oxime Formation at Neutral pH

    PubMed Central

    2015-01-01

    The discovery of two new classes of catalysts for hydrazone and oxime formation in water at neutral pH, namely 2-aminophenols and 2-(aminomethyl)benzimidazoles, is reported. Kinetics studies in aqueous solutions at pH 7.4 revealed rate enhancements up to 7-fold greater than with classic aniline catalysis. 2-(Aminomethyl)benzimidazoles were found to be effective catalysts with otherwise challenging aryl ketone substrates. PMID:25545888

  1. Catalytic Activity of Cationic and Neutral Silver(I)-XPhos Complexes with Nitrogen Ligands or Tolylsulfonate for Mannich and Aza-Diels-Alder Coupling Reactions.

    PubMed

    Grirrane, Abdessamad; Álvarez, Eleuterio; García, Hermenegildo; Corma, Avelino

    2016-01-01

    Cationic and neutral silver(I)-L complexes (L=Buchwald-type biaryl phosphanes) with nitrogen co-ligands or organosulfonate counter ions have been synthesised and characterised through their structural and spectroscopic properties. At room temperature, both cationic and neutral silver(I)-L complexes are extremely active catalysts in the promotion of the single and double A(3) coupling of terminal (di)alkynes, pyrrolidine and formaldehyde. In addition, the aza-Diels-Alder two- and three-component coupling reactions of Danishefsky's diene with an imine or amine and aldehyde are efficiently catalysed by these cationic or neutral silver(I)-L complexes. The solvent influences the catalytic performance due to limited complex solubility or solvent decomposition and reactivity. The isolation of new silver(I)-L complexes with reagents as ligands lends support to mechanistic proposals for such catalytic processes. The activity, stability and metal-distal arene interaction of these silver(I)-L catalysts have been compared with those of analogous cationic gold(I) and copper(I) complexes. PMID:26598792

  2. Ultrasensitive Analysis of Binding Affinity of HIV Receptor and Neutralizing Antibody Using Solution-Phase Electrochemiluminescence Assay

    PubMed Central

    Xu, Xiao-Hong Nancy; Wen, Zhaoyang; Brownlow, William J.

    2012-01-01

    Binding of a few ligand molecules with its receptors on cell surface can initiate cellular signaling transduction pathways, and trigger viral infection of host cells. HIV-1 infects host T-cells by binding its viral envelope protein (gp120) with its receptor (a glycoprotein, CD4) on T cells. Primary strategies to prevent and treat HIV infection is to develop therapies (e.g., neutralizing antibodies) that can block specific binding of CD4 with gp120. The infection often leads to the lower counts of CD4 cells, which makes it an effective biomarker to monitor the AIDS progression and treatment. Despite research over decades, quantitative assays for effective measurements of binding affinities of protein-protein (ligand-receptor, antigen-antibody) interactions remains highly sought. Solid-phase electrochemiluminescence (ECL) immunoassay has been commonly used to capture analytes from the solution for analysis, which involves immobilization of antibody on solid surfaces (micron-sized beads), but it cannot quantitatively measure binding affinities of molecular interactions. In this study, we have developed solution-phase ECL assay with a wide dynamic range (0–2 nM) and high sensitivity and specificity for quantitative analysis of CD4 at femtomolar level and their binding affinity with gp120 and monoclonal antibodies (MABs). We found that binding affinities of CD4 with gp120 and MAB (Q4120) are 9.5×108 and 1.2×109 M−1, respectively. The results also show that MAB (Q4120) of CD4 can completely block the binding of gp120 with CD4, while MAB (17b) of gp120 can only partially block their interaction. This study demonstrates that the solution-phase ECL assay can be used for ultrasensitive and quantitative analysis of binding affinities of protein-protein interactions in solution for better understating of protein functions and identification of effective therapies to block their interactions. PMID:23565071

  3. Double C-H bond activation of hydrocarbons by a gas phase neutral oxide cluster: the importance of spin state.

    PubMed

    Wang, Zhe-Chen; Yin, Shi; Bernstein, Elliot R

    2013-03-21

    The neutral cluster V2O5 is generated and detected in the gas phase. Its reactivity toward butane is studied both experimentally and theoretically. Experimental results show clearly that neutral V2O5 can react with n-butane (C4H10) to generate V2O5H2, indicating double hydrogen atom transfer from C4H10 to V2O5 to produce C4H8. Further experimental evidence indicates that V2O5 is only partially reacted even at very high concentrations of C4H10. Density functional theory (DFT) studies show that the lowest energy triplet state of V2O5 is reactive toward C4H10, whereas the ground state singlet V2O5 is inert. Calculated results are in agreement with experimental findings, and a detailed reaction mechanism is provided. Reactions of V2O5H2 with several oxidants are also studied theoretically to find a path to regenerate V2O5. Neutral (3)V2O5 can also react with C2H6 to form V2O5H2 and C2H4, but only as a minor reaction channel; the major product is the adsorption product V2O5(C2H6). PMID:23441829

  4. Crystallization and preliminary X-ray crystallographic analysis of the sclerostin-neutralizing Fab AbD09097.

    PubMed

    Boschert, Verena; Muth, Eva Maria; Knappik, Achim; Frisch, Christian; Mueller, Thomas D

    2015-04-01

    The secreted cystine-knot protein sclerostin was first identified from genetic screening of patients suffering from the rare bone-overgrowth diseases sclerosteosis and van Buchem disease. Sclerostin acts a negative regulator of bone growth through inhibiting the canonical Wnt signalling cascade by binding to and blocking the Wnt co-receptor LRP5/6. Its function in blocking osteoblastogenesis makes it an important target for osteoanabolic therapy approaches to treat osteoporosis, which is characterized by a progressive decrease in bone mass and density. In this work, the production, crystallization and preliminary X-ray diffraction data analysis of a sclerostin-neutralizing human Fab antibody fragment, AbD09097, obtained from a naive antibody library are reported. Crystals of the Fab AbD09097 belonged to space group P21, with unit-cell parameters a = 45.19, b = 78.49, c = 59.20 Å, β = 95.71° and diffracted X-rays to a resolution of 1.8 Å.

  5. Analysis of cross-reactive neutralizing antibodies in human HFMD serum with an EV71 pseudovirus-based assay.

    PubMed

    Zhang, Huafei; An, Dong; Liu, Wei; Mao, Qunying; Jin, Jun; Xu, Lin; Sun, Shiyang; Jiang, Liping; Li, Xiaojun; Shao, Jie; Ma, Hongxia; Huang, Xueyong; Guo, Shijie; Chen, Haiying; Cheng, Tong; Yang, Lisheng; Su, Weiheng; Kong, Wei; Liang, Zhenglun; Jiang, Chunlai

    2014-01-01

    Hand, foot and mouth disease, associated with enterovirus 71 (EV71) infections, has recently become an important public health issue throughout the world. Serum neutralizing antibodies are major indicators of EV71 infection and protective immunity. However, the potential for cross-reactivity of neutralizing antibodies for different EV71 genotypes and subgenotypes is unclear. Here we measured the cross-reactive neutralizing antibody titers against EV71 of different genotypes or subgenotypes in sera collected from EV71-infected children and vaccine-inoculated children in a phase III clinical trial (ClinicalTrials.gov Identifier: NCT01636245) using a new pseudovirus-based neutralization assay. Antibodies induced by EV71-C4a were cross-reactive for different EV71 genotypes, demonstrating that C4a is a good candidate strain for an EV71 vaccine. Our study also demonstrated that this new assay is practical for analyses of clinical samples from epidemiological and vaccine studies. PMID:24964084

  6. Analysis of Cross-Reactive Neutralizing Antibodies in Human HFMD Serum with an EV71 Pseudovirus-Based Assay

    PubMed Central

    Jin, Jun; Xu, Lin; Sun, Shiyang; Jiang, Liping; Li, Xiaojun; Shao, Jie; Ma, Hongxia; Huang, Xueyong; Guo, Shijie; Chen, Haiying; Cheng, Tong; Yang, Lisheng; Su, Weiheng; Kong, Wei; Liang, Zhenglun; Jiang, Chunlai

    2014-01-01

    Hand, foot and mouth disease, associated with enterovirus 71 (EV71) infections, has recently become an important public health issue throughout the world. Serum neutralizing antibodies are major indicators of EV71 infection and protective immunity. However, the potential for cross-reactivity of neutralizing antibodies for different EV71 genotypes and subgenotypes is unclear. Here we measured the cross-reactive neutralizing antibody titers against EV71 of different genotypes or subgenotypes in sera collected from EV71-infected children and vaccine-inoculated children in a phase III clinical trial (ClinicalTrials.gov Identifier: NCT01636245) using a new pseudovirus-based neutralization assay. Antibodies induced by EV71-C4a were cross-reactive for different EV71 genotypes, demonstrating that C4a is a good candidate strain for an EV71 vaccine. Our study also demonstrated that this new assay is practical for analyses of clinical samples from epidemiological and vaccine studies. PMID:24964084

  7. Analysis of cross-reactive neutralizing antibodies in human HFMD serum with an EV71 pseudovirus-based assay.

    PubMed

    Zhang, Huafei; An, Dong; Liu, Wei; Mao, Qunying; Jin, Jun; Xu, Lin; Sun, Shiyang; Jiang, Liping; Li, Xiaojun; Shao, Jie; Ma, Hongxia; Huang, Xueyong; Guo, Shijie; Chen, Haiying; Cheng, Tong; Yang, Lisheng; Su, Weiheng; Kong, Wei; Liang, Zhenglun; Jiang, Chunlai

    2014-01-01

    Hand, foot and mouth disease, associated with enterovirus 71 (EV71) infections, has recently become an important public health issue throughout the world. Serum neutralizing antibodies are major indicators of EV71 infection and protective immunity. However, the potential for cross-reactivity of neutralizing antibodies for different EV71 genotypes and subgenotypes is unclear. Here we measured the cross-reactive neutralizing antibody titers against EV71 of different genotypes or subgenotypes in sera collected from EV71-infected children and vaccine-inoculated children in a phase III clinical trial (ClinicalTrials.gov Identifier: NCT01636245) using a new pseudovirus-based neutralization assay. Antibodies induced by EV71-C4a were cross-reactive for different EV71 genotypes, demonstrating that C4a is a good candidate strain for an EV71 vaccine. Our study also demonstrated that this new assay is practical for analyses of clinical samples from epidemiological and vaccine studies.

  8. Purification and some physico-chemical and enzymic properties of a calcium ion-activated neutral proteinase from rabbit skeletal muscle

    PubMed Central

    Azanza, Jean-Louis; Raymond, Jacques; Robin, Jean-Michel; Cottin, Patrick; Ducastaing, André

    1979-01-01

    Ca2+-activated neutral proteinase was purified from rabbit skeletal muscle by a method involving DEAE-Sephacel chromatography, affinity chromatography on organomercurial–Sepharose and gel filtration on Sephacryl S-200 and Sephadex G-150. The SDS (sodium dodecyl sulphate)/polyacrylamide-gel-electrophoresis data show that the purified enzyme contains only one polypeptide chain of mol.wt. 73000. The purification procedure used allowed us to eliminate a contaminant containing two components of mol.wt. about 30000 each. Whole casein or α1-casein were hydrolysed with a maximum rate at 30°C, pH7.5, and with 5mm-CaCl2, but myofibrils were found to be a very susceptible substrate for this proteinase. This activity is associated with the destruction of the Z-discs, which is caused by the solubilization of the Z-line proteins. The activity of the proteinase in vitro is not limited to the removal of Z-line. SDS/polyacrylamide-gel electrophoresis on larger plates showed the ability of the proteinase to degrade myofibrils more extensively than previously supposed. This proteolysis resulted in the production of a 30000-dalton component as well as in various other higher- and lower-molecular-weight peptide fragments. Troponin T, troponin I, α-tropomyosin, some high-molecular-weight proteins (M protein, heavy chain of myosin) and three unidentified proteins are degraded. Thus the number of proteinase-sensitive regions in the myofibrils is greater than as previously reported by Dayton, Goll, Zeece, Robson & Reville [(1976) Biochemistry 15, 2150–2158]. The Ca2+-activated neutral proteinase is not a chymotrypsin- or trypsin-like enzyme, but it reacted with all the classic thiol-proteinase inhibitors for cathepsin B, papain, bromelain and ficin. Thus the proteinase was proved to have an essential thiol group. Antipain and leupeptin are also inhibitors of the Ca2+-activated neutral proteinase. ImagesFig. 1.Fig. 2.Fig. 3. PMID:534501

  9. Structural Characterization of Humanized Nanobodies with Neutralizing Activity against the Bordetella pertussis CyaA-Hemolysin: Implications for a Potential Epitope of Toxin-Protective Antigen

    PubMed Central

    Malik, Aijaz Ahmad; Imtong, Chompounoot; Sookrung, Nitat; Katzenmeier, Gerd; Chaicumpa, Wanpen; Angsuthanasombat, Chanan

    2016-01-01

    Previously, the 126-kDa CyaA-hemolysin (CyaA-Hly) fragment cloned from Bordetella pertussis—the causative agent of whooping cough—and functionally expressed in Escherichia coli was revealed as a key determinant for CyaA-mediated hemolysis against target erythrocytes. Here, phagemid-transfected E. coli clones producing nanobodies capable of binding to CyaA-Hly were selected from a humanized-camel VH/VHH phage-display library. Subsequently verified for binding activities by indirect ELISA and Western blotting, four CyaA-Hly-specific nanobodies were obtained and designated according to the presence/absence of VHH-hallmark amino acids as VHH2, VH5, VH18 and VHH37. In vitro neutralization assay revealed that all four ~17-kDa His-tagged VH/VHH nanobodies, in particular VHH37, which were over-expressed as inclusions and successfully unfolded-refolded, were able to effectively inhibit CyaA-Hly-mediated hemolysis. Phage-mimotope searching revealed that only peptides with sequence homologous to Linker 1 connecting Blocks I and II within the CyaA-RTX subdomain were able to bind to these four CyaA-Hly-specific nanobodies. Structural analysis of VHH37 via homology modeling and intermolecular docking confirmed that this humanized nanobody directly interacts with CyaA-RTX/Linker 1 through multiple hydrogen and ionic bonds. Altogether, our present data demonstrate that CyaA-RTX/Linker 1 could serve as a potential epitope of CyaA-protective antigen that may be useful for development of peptide-based pertussis vaccines. Additionally, such toxin-specific nanobodies have a potential for test-driven development of a ready-to-use therapeutic in passive immunization for mitigation of disease severity. PMID:27043627

  10. Structural Characterization of Humanized Nanobodies with Neutralizing Activity against the Bordetella pertussis CyaA-Hemolysin: Implications for a Potential Epitope of Toxin-Protective Antigen.

    PubMed

    Malik, Aijaz Ahmad; Imtong, Chompounoot; Sookrung, Nitat; Katzenmeier, Gerd; Chaicumpa, Wanpen; Angsuthanasombat, Chanan

    2016-04-01

    Previously, the 126-kDa CyaA-hemolysin (CyaA-Hly) fragment cloned from Bordetella pertussis--the causative agent of whooping cough--and functionally expressed in Escherichia coli was revealed as a key determinant for CyaA-mediated hemolysis against target erythrocytes. Here, phagemid-transfected E. coli clones producing nanobodies capable of binding to CyaA-Hly were selected from a humanized-camel VH/VHH phage-display library. Subsequently verified for binding activities by indirect ELISA and Western blotting, four CyaA-Hly-specific nanobodies were obtained and designated according to the presence/absence of VHH-hallmark amino acids as VHH2, VH5, VH18 and VHH37. In vitro neutralization assay revealed that all four ~17-kDa His-tagged VH/VHH nanobodies, in particular VHH37, which were over-expressed as inclusions and successfully unfolded-refolded, were able to effectively inhibit CyaA-Hly-mediated hemolysis. Phage-mimotope searching revealed that only peptides with sequence homologous to Linker 1 connecting Blocks I and II within the CyaA-RTX subdomain were able to bind to these four CyaA-Hly-specific nanobodies. Structural analysis of VHH37 via homology modeling and intermolecular docking confirmed that this humanized nanobody directly interacts with CyaA-RTX/Linker 1 through multiple hydrogen and ionic bonds. Altogether, our present data demonstrate that CyaA-RTX/Linker 1 could serve as a potential epitope of CyaA-protective antigen that may be useful for development of peptide-based pertussis vaccines. Additionally, such toxin-specific nanobodies have a potential for test-driven development of a ready-to-use therapeutic in passive immunization for mitigation of disease severity.

  11. Structural Characterization of Humanized Nanobodies with Neutralizing Activity against the Bordetella pertussis CyaA-Hemolysin: Implications for a Potential Epitope of Toxin-Protective Antigen.

    PubMed

    Malik, Aijaz Ahmad; Imtong, Chompounoot; Sookrung, Nitat; Katzenmeier, Gerd; Chaicumpa, Wanpen; Angsuthanasombat, Chanan

    2016-04-01

    Previously, the 126-kDa CyaA-hemolysin (CyaA-Hly) fragment cloned from Bordetella pertussis--the causative agent of whooping cough--and functionally expressed in Escherichia coli was revealed as a key determinant for CyaA-mediated hemolysis against target erythrocytes. Here, phagemid-transfected E. coli clones producing nanobodies capable of binding to CyaA-Hly were selected from a humanized-camel VH/VHH phage-display library. Subsequently verified for binding activities by indirect ELISA and Western blotting, four CyaA-Hly-specific nanobodies were obtained and designated according to the presence/absence of VHH-hallmark amino acids as VHH2, VH5, VH18 and VHH37. In vitro neutralization assay revealed that all four ~17-kDa His-tagged VH/VHH nanobodies, in particular VHH37, which were over-expressed as inclusions and successfully unfolded-refolded, were able to effectively inhibit CyaA-Hly-mediated hemolysis. Phage-mimotope searching revealed that only peptides with sequence homologous to Linker 1 connecting Blocks I and II within the CyaA-RTX subdomain were able to bind to these four CyaA-Hly-specific nanobodies. Structural analysis of VHH37 via homology modeling and intermolecular docking confirmed that this humanized nanobody directly interacts with CyaA-RTX/Linker 1 through multiple hydrogen and ionic bonds. Altogether, our present data demonstrate that CyaA-RTX/Linker 1 could serve as a potential epitope of CyaA-protective antigen that may be useful for development of peptide-based pertussis vaccines. Additionally, such toxin-specific nanobodies have a potential for test-driven development of a ready-to-use therapeutic in passive immunization for mitigation of disease severity. PMID:27043627

  12. Neutral Buoyancy Simulator Test

    NASA Technical Reports Server (NTRS)

    1995-01-01

    A diver tests a secondary camera and maneuvering platform in Marshall's Neutral Buoyancy Simulator (NBS).The secondary camera will be beneficial for recording repairs and other extra vehicular activities (EVA) the astronuats will perform while making repairs on the Hubble Space Telescope (HST). The maneuvering platform was developed to give the astronauts something to stand on while performing maintenance tasks. These platforms were developed to be mobile so that the astronauts could move them to accommadate different sites.

  13. Charge-exchange measurements of MHD activity during neutral beam injection in the Princeton Large Torus and the Poloidal Divertor Experiment

    SciTech Connect

    Goldston, R.J.; Kaita, R.; Beiersdorfer, P.; Gammel, G.; Herndon, D.L.; McCune, D.C.; Meyerhofer, D.D.

    1986-07-01

    The horizontally scanning, multiangle charge-exchange analyzers on the Princeton Large Torus (PLT) and the Poloidal Divertor Experiment (PDX) were used to study the effects of MHD activity on the background ion distribution function and on the beam ion slowing-down process during high-power neutral injection. Sawtooth oscillations were observed in the fast ion flux on PLT and PDX, and measurements with neutral beams providing local neutral density enhancement indicate that ions are transported radially when these events occur. With near-perpendicular injection in PDX, at the lower toroidal fields necessary to maximize beta, rapid, repetitive bursts of greatly enhanced charge-exchange flux were observed. These are associated with the ''fishbone'' MHD instability, and a substantial depletion of the perpendicular slowing-down spectrum below the injection energy was seen. A simple phenomenological model for this loss mechanism was developed, and its use in simulation codes has been successful in providing good agreement with the data. The behavior and characteristics of this model are well matched by the direct theoretical calculations.

  14. US--ITER activation analysis

    SciTech Connect

    Attaya, H.; Gohar, Y.; Smith, D.

    1990-09-01

    Activation analysis has been made for the US ITER design. The radioactivity and the decay heat have been calculated, during operation and after shutdown for the two ITER phases, the Physics Phase and the Technology Phase. The Physics Phase operates about 24 full power days (FPDs) at fusion power level of 1100 MW and the Technology Phase has 860 MW fusion power and operates for about 1360 FPDs. The point-wise gamma sources have been calculated everywhere in the reactor at several times after shutdown of the two phases and are then used to calculate the biological dose everywhere in the reactor. Activation calculations have been made also for ITER divertor. The results are presented for different continuous operation times and for only one pulse. The effect of the pulsed operation on the radioactivity is analyzed. 6 refs., 12 figs., 1 tab.

  15. Prompt-gamma activation analysis

    SciTech Connect

    Lindstrom, R.M.

    1993-01-01

    A permenent, full-time instrument for prompt-gamma activation analysis is nearing completion as part of the Cold Neutron Research Facility (CNRF). The design of the analytical system has been optimized for high gamma detection efficiency and low background, particularly for hydrogen. Because of the purity of the neutron beam, shielding requirements are modest and the scatter-capture background is low. As a result of a compact sample-detector geometry, the sensitivity (counting rate per gram of analyte) is a factor of four better than the existing Maryland-NIST thermal-neutron instrument at the reactor. Hydrogen backgrounds of a few micrograms have already been achieved, which promises to be of value in numerous applications where quantitative nondestructive analysis of small quantities of hydrogen in materials is necessary.

  16. Iron-Carbonyl-Catalyzed Redox-Neutral [4+2] Annulation of N-H Imines and Internal Alkynes by C-H Bond Activation.

    PubMed

    Jia, Teng; Zhao, Chongyang; He, Ruoyu; Chen, Hui; Wang, Congyang

    2016-04-18

    Stoichiometric C-H bond activation of arenes mediated by iron carbonyls was reported by Pauson as early as in 1965, yet the catalytic C-H transformations have not been developed. Herein, an iron-catalyzed annulation of N-H imines and internal alkynes to furnish cis-3,4-dihydroisoquinolines is described, and represents the first iron-carbonyl-catalyzed C-H activation reaction of arenes. Remarkablely, this is also the first redox-neutral [4+2] annulation of imines and alkynes proceeding by C-H activation. The reaction also features only cis stereoselectivity and excellent atom economy as neither base, nor external ligand, nor additive is required. Experimental and theoretical studies reveal an oxidative addition mechanism for C-H bond activation to afford a dinuclear ferracycle and a synergetic diiron-promoted H-transfer to the alkyne as the turnover-determining step.

  17. Experimental and DFT Studies Explain Solvent Control of C-H Activation and Product Selectivity in the Rh(III)-Catalyzed Formation of Neutral and Cationic Heterocycles.

    PubMed

    Davies, David L; Ellul, Charles E; Macgregor, Stuart A; McMullin, Claire L; Singh, Kuldip

    2015-08-01

    A range of novel heterocyclic cations have been synthesized by the Rh(III)-catalyzed oxidative C-N and C-C coupling of 1-phenylpyrazole, 2-phenylpyridine, and 2-vinylpyridine with alkynes (4-octyne and diphenylacetylene). The reactions proceed via initial C-H activation, alkyne insertion, and reductive coupling, and all three of these steps are sensitive to the substrates involved and the reaction conditions. Density functional theory (DFT) calculations show that C-H activation can proceed via a heteroatom-directed process that involves displacement of acetate by the neutral substrate to form charged intermediates. This step (which leads to cationic C-N coupled products) is therefore favored by more polar solvents. An alternative non-directed C-H activation is also possible that does not involve acetate displacement and so becomes favored in low polarity solvents, leading to C-C coupled products. Alkyne insertion is generally more favorable for diphenylacetylene over 4-octyne, but the reverse is true of the reductive coupling step. The diphenylacetylene moiety can also stabilize unsaturated seven-membered rhodacycle intermediates through extra interaction with one of the Ph substituents. With 1-phenylpyrazole this effect is sufficient to suppress the final C-N reductive coupling. A comparison of a series of seven-membered rhodacycles indicates the barrier to coupling is highly sensitive to the two groups involved and follows the trend C-N(+) > C-N > C-C (i.e., involving the formation of cationic C-N, neutral C-N, and neutral C-C coupled products, respectively). PMID:26115418

  18. Acidic extracellular pH neutralizes the autophagy-inhibiting activity of chloroquine: implications for cancer therapies.

    PubMed

    Pellegrini, Paola; Strambi, Angela; Zipoli, Chiara; Hägg-Olofsson, Maria; Buoncervello, Maria; Linder, Stig; De Milito, Angelo

    2014-04-01

    Acidic pH is an important feature of tumor microenvironment and a major determinant of tumor progression. We reported that cancer cells upregulate autophagy as a survival mechanism to acidic stress. Inhibition of autophagy by administration of chloroquine (CQ) in combination anticancer therapies is currently evaluated in clinical trials. We observed in 3 different human cancer cell lines cultured at acidic pH that autophagic flux is not blocked by CQ. This was consistent with a complete resistance to CQ toxicity in cells cultured in acidic conditions. Conversely, the autophagy-inhibiting activity of Lys-01, a novel CQ derivative, was still detectable at low pH. The lack of CQ activity was likely dependent on a dramatically reduced cellular uptake at acidic pH. Using cell lines stably adapted to chronic acidosis we could confirm that CQ lack of activity was merely caused by acidic pH. Moreover, unlike CQ, Lys-01 was able to kill low pH-adapted cell lines, although higher concentrations were required as compared with cells cultured at normal pH conditions. Notably, buffering medium pH in low pH-adapted cell lines reverted CQ resistance. In vivo analysis of tumors treated with CQ showed that accumulation of strong LC3 signals was observed only in normoxic areas but not in hypoxic/acidic regions. Our observations suggest that targeting autophagy in the tumor environment by CQ may be limited to well-perfused regions but not achieved in acidic regions, predicting possible limitations in efficacy of CQ in antitumor therapies. PMID:24492472

  19. Second generation of pseudotype-based serum neutralization assay for Nipah virus antibodies: sensitive and high-throughput analysis utilizing secreted alkaline phosphatase.

    PubMed

    Kaku, Yoshihiro; Noguchi, Akira; Marsh, Glenn A; Barr, Jennifer A; Okutani, Akiko; Hotta, Kozue; Bazartseren, Boldbaatar; Fukushi, Shuetsu; Broder, Christopher C; Yamada, Akio; Inoue, Satoshi; Wang, Lin-Fa

    2012-01-01

    Nipah virus (NiV), Paramyxoviridae, Henipavirus, is classified as a biosafety level (BSL) 4 pathogen, along with the closely related Hendra virus (HeV). A novel serum neutralization test was developed for measuring NiV neutralizing antibodies under BSL2 conditions using a recombinant vesicular stomatitis virus (VSV) expressing secreted alkaline phosphatase (SEAP) and pseudotyped with NiV F/G proteins (VSV-NiV-SEAP). A unique characteristic of this novel assay is the ability to obtain neutralization titers by measuring SEAP activity in supernatant using a common ELISA plate reader. This confers a remarkable advantage over the first generation of NiV-pseudotypes expressing green fluorescent protein or luciferase, which require expensive and specific measuring equipment. Using panels of NiV- and HeV-specific sera from various species, the VSV-NiV-SEAP assay demonstrated neutralizing antibody status (positive/negative) consistent with that obtained by conventional live NiV test, and gave higher antibody titers than the latter. Additionally, when screening sixty-six fruit bat sera at one dilution, the VSV-NiV-SEAP assay produced identical results to the live NiV test and only required a very small amount (2μl) of sera. The results suggest that this novel VSV-NiV-SEAP assay is safe, useful for high-throughput screening of sera using an ELISA plate reader, and has high sensitivity and specificity. PMID:22115786

  20. Neutralization of (NK-cell-derived) B-cell activating factor by Belimumab restores sensitivity of chronic lymphoid leukemia cells to direct and Rituximab-induced NK lysis.

    PubMed

    Wild, J; Schmiedel, B J; Maurer, A; Raab, S; Prokop, L; Stevanović, S; Dörfel, D; Schneider, P; Salih, H R

    2015-08-01

    Natural killer (NK) cells are cytotoxic lymphocytes that substantially contribute to the therapeutic benefit of antitumor antibodies like Rituximab, a crucial component in the treatment of B-cell malignancies. In chronic lymphocytic leukemia (CLL), the ability of NK cells to lyse the malignant cells and to mediate antibody-dependent cellular cytotoxicity upon Fc receptor stimulation is compromised, but the underlying mechanisms are largely unclear. We report here that NK-cells activation-dependently produce the tumor necrosis factor family member 'B-cell activating factor' (BAFF) in soluble form with no detectable surface expression, also in response to Fc receptor triggering by therapeutic CD20-antibodies. BAFF in turn enhanced the metabolic activity of primary CLL cells and impaired direct and Rituximab-induced lysis of CLL cells without affecting NK reactivity per se. The neutralizing BAFF antibody Belimumab, which is approved for treatment of systemic lupus erythematosus, prevented the effects of BAFF on the metabolism of CLL cells and restored their susceptibility to direct and Rituximab-induced NK-cell killing in allogeneic and autologous experimental systems. Our findings unravel the involvement of BAFF in the resistance of CLL cells to NK-cell antitumor immunity and Rituximab treatment and point to a benefit of combinatory approaches employing BAFF-neutralizing drugs in B-cell malignancies.

  1. Large-Scale Analysis of B-Cell Epitopes on Influenza Virus Hemagglutinin – Implications for Cross-Reactivity of Neutralizing Antibodies

    PubMed Central

    Sun, Jing; Kudahl, Ulrich J.; Simon, Christian; Cao, Zhiwei; Reinherz, Ellis L.; Brusic, Vladimir

    2014-01-01

    Influenza viruses continue to cause substantial morbidity and mortality worldwide. Fast gene mutation on surface proteins of influenza virus result in increasing resistance to current vaccines and available antiviral drugs. Broadly neutralizing antibodies (bnAbs) represent targets for prophylactic and therapeutic treatments of influenza. We performed a systematic bioinformatics study of cross-reactivity of neutralizing antibodies (nAbs) against influenza virus surface glycoprotein hemagglutinin (HA). This study utilized the available crystal structures of HA complexed with the antibodies for the analysis of tens of thousands of HA sequences. The detailed description of B-cell epitopes, measurement of epitope area similarity among different strains, and estimation of antibody neutralizing coverage provide insights into cross-reactivity status of existing nAbs against influenza virus. We have developed a method to assess the likely cross-reactivity potential of bnAbs for influenza strains, either newly emerged or existing. Our method catalogs influenza strains by a new concept named discontinuous peptide, and then provide assessment of cross-reactivity. Potentially cross-reactive strains are those that share 100% identity with experimentally verified neutralized strains. By cataloging influenza strains and their B-cell epitopes for known bnAbs, our method provides guidance for selection of representative strains for further experimental design. The knowledge of sequences, their B-cell epitopes, and differences between historical influenza strains, we enhance our preparedness and the ability to respond to the emerging pandemic threats. PMID:24570677

  2. Comprehensive analysis of sperm DNA fragmentation by five different assays: TUNEL assay, SCSA, SCD test and alkaline and neutral Comet assay.

    PubMed

    Ribas-Maynou, J; García-Peiró, A; Fernández-Encinas, A; Abad, C; Amengual, M J; Prada, E; Navarro, J; Benet, J

    2013-09-01

    Sperm DNA fragmentation (SDF) is becoming an important test to assess male infertility. Several different tests are available, but no consensus has yet been reached as to which tests are most predictive of infertility. Few publications have reported a comprehensive analysis comparing these methods within the same population. The objective of this study was to analyze the differences between the five most common methodologies, to study their correlations and to establish their cut-off values, sensitivity and specificity in predicting male infertility. We found differences in SDF between fertile donors and infertile patients in TUNEL, SCSA, SCD and alkaline Comet assays, but none with the neutral Comet assay. The alkaline COMET assay was the best in predicting male infertility followed by TUNEL, SCD and SCSA, whereas the neutral COMET assay had no predictive power. For our patient population, threshold values for infertility were 20.05% for TUNEL assay, 18.90% for SCSA, 22.75% for the SCD test, 45.37% for alkaline Comet and 34.37% for neutral Comet. This work establishes in a comprehensive study that the all techniques except neutral Comet are useful to distinguish fertile and infertile men.

  3. Neutral current interactions in MINOS

    SciTech Connect

    Sousa, Alexandre; /Oxford U.

    2007-07-01

    The Main Injector Neutrino Oscillation Search (MINOS) long-baseline experiment has been actively collecting beam data since 2005, having already accumulated 3 x 10{sup 20} protons-on-target (POT). The several million neutrinos per year observed at the Near detector may improve the existing body of knowledge of neutrino cross-sections and the Near-Far comparison of the observed energy spectrum neutral current events constrains oscillations into sterile neutrinos. MINOS capabilities of observing neutral current neutrino events are described and the employed methodology for event selection is discussed, along with preliminary results obtained. An outlook on the expected neutral current related contributions from MINOS is also presented.

  4. An immunogen containing four tandem 10E8 epitope repeats with exposed key residues induces antibodies that neutralize HIV-1 and activates an ADCC reporter gene.

    PubMed

    Sun, Zhiwu; Zhu, Yun; Wang, Qian; Ye, Ling; Dai, Yanyan; Su, Shan; Yu, Fei; Ying, Tianlei; Yang, Chinglai; Jiang, Shibo; Lu, Lu

    2016-01-01

    After three decades of intensive research efforts, an effective vaccine against HIV-1 remains to be developed. Several broadly neutralizing antibodies to HIV-1, such as 10E8, recognize the membrane proximal external region (MPER) of the HIV-1 gp41 protein. Thus, the MPER is considered to be a very important target for vaccine design. However, the MPER segment has very weak immunogenicity and tends to insert its epitope residues into the cell membrane, thereby avoiding antibody binding. To address this complication in vaccine development, we herein designed a peptide, designated 10E8-4P, containing four copies of the 10E8 epitope as an immunogen. As predicted by structural simulation, 10E8-4P exhibits a well-arranged tandem helical conformation, with the key residues in the 10E8 epitope oriented at different angles, thus suggesting that some of these key residues may be exposed outside of the lipid membrane. Compared with a peptide containing a single 10E8 epitope (10E8-1P), 10E8-4P not only exhibited better antigenicity but also elicited neutralizing antibody response against HIV-1 pseudoviruses, whereas 10E8-1P could not induce detectable neutralizing antibody response. Importantly, antibodies elicited by 10E8-4P also possessed a strong ability to activate an antibody-dependent cell-mediated cytotoxicity (ADCC) reporter gene, thus suggesting that they may have ADCC activity. Therefore, this strategy shows promise for further optimization and application in future HIV-1 vaccine design. PMID:27329850

  5. MSFC Skylab neutral buoyancy simulator

    NASA Technical Reports Server (NTRS)

    1974-01-01

    The use of a neutral buoyancy simulator for developing extravehicular activity systems and for training astronauts in weightless activities is discussed. The construction of the facility and the operations are described. The types of tests and the training activities conducted in the simulator are reported. Photographs of the components of the simulator and actual training exercises are included.

  6. Quantifying Ruminal Digestion of Organic Matter and Neutral Detergent Fiber Using Omasal Sampling in Cattle--A Meta-Analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A data set from 32 studies (122 diets) was used to evaluate the accuracy and precision of the omasal sampling technique by investigating the relationships between ruminal and total digestion of neutral detergent fiber (NDF), between intake and apparent and true ruminal digestion of organic matter (O...

  7. Analysis of Physical Science Textbooks for Conceptual Frameworks on Acids, Bases and Neutralization: Implications for Students' Conceptual Understanding.

    ERIC Educational Resources Information Center

    Erduran, Sibel

    Eight physical science textbooks were analyzed for coverage on acids, bases, and neutralization. At the level of the text, clarity and coherence of statements were investigated. The conceptual framework for this topic was represented in a concept map which was used as a coding tool for tracing concepts and links present in textbooks. Cognitive…

  8. A constant threat for HIV: Fc-engineering to enhance broadly neutralizing antibody activity for immunotherapy of the acquired immunodeficiency syndrome.

    PubMed

    Nimmerjahn, Falk

    2015-08-01

    Passive immunotherapy with polyclonal or hyperimmune serum immunoglobulin G (IgG) preparations provides an efficient means of protecting immunocompromised patients from microbial infections. More recently, the use of passive immunotherapy to prevent or to treat established infections with the human immunodeficiency virus (HIV) has gained much attention, due to promising preclinical data obtained in monkey and humanized mouse in vivo model systems, demonstrating that the transfer of HIV-specific antibodies can not only prevent HIV infection, but also diminish virus load during chronic infection. Furthermore, an array of broadly neutralizing HIV-specific antibodies has become available and the importance of the IgG constant region as a critical modulator of broadly neutralizing activity has been demonstrated. The aim of this review is to summarize the most recent findings with regard to the molecular and cellular mechanisms responsible for antibody-mediated clearance of HIV infection, and to discuss how this may help to improve HIV therapy via optimizing Fcγ-receptor-dependent activities of HIV-specific antibodies.

  9. Neutral and ionic platinum compounds containing a cyclometallated chiral primary amine: synthesis, antitumor activity, DNA interaction and topoisomerase I-cathepsin B inhibition.

    PubMed

    Albert, Joan; Bosque, Ramon; Crespo, Margarita; Granell, Jaume; López, Concepción; Martín, Raquel; González, Asensio; Jayaraman, Anusha; Quirante, Josefina; Calvis, Carme; Badía, Josefa; Baldomà, Laura; Font-Bardia, Mercè; Cascante, Marta; Messeguer, Ramon

    2015-08-14

    The synthesis and preliminary biological evaluation of neutral and cationic platinum derivatives of chiral 1-(1-naphthyl)ethylamine are reported, namely cycloplatinated neutral complexes [PtCl{(R or S)-NH(2)CH(CH(3))C(10)H(6)}(L)] [L = SOMe(2) ( 1-R or 1-S ), L = PPh(3) (2-R or 2-S), L = P(4-FC(6)H(4))(3) (3-R), L = P(CH(2))(3)N(3)(CH(2))(3) (4-R)], cycloplatinated cationic complexes [Pt{(R)-NH(2)CH(CH(3))C(10)H(6)}{L}]Cl [L = Ph(2)PCH(2)CH(2)PPh(2) (5-R), L = (C(6)F(5))(2)PCH(2)CH(2)P(C(6)F(5))(2) (6-R)] and the Pt(ii) coordination compound trans-[PtCl(2){(R)-NH(2)CH(CH(3))C(10)H(6)}(2)] (7-R). The X-ray molecular structure of 7-R is reported. The cytotoxic activity against a panel of human adenocarcinoma cell lines (A-549 lung, MDA-MB-231 and MCF-7 breast, and HCT-116 colon), cell cycle arrest and apoptosis, DNA interaction, topoisomerase I and cathepsin B inhibition, and Pt cell uptake of the studied compounds are presented. Remarkable cytotoxicity was observed for most of the synthesized Pt(ii) compounds regardless of (i) the absolute configuration R or S, and (ii) the coordinated/cyclometallated (neutral or cationic) nature of the complexes. The most potent compound 2-R (IC(50) = 270 nM) showed a 148-fold increase in potency with regard to cisplatin in HCT-116 colon cancer cells. Preliminary biological results point out to different biomolecular targets for the investigated compounds. Neutral cyclometallated complexes 1-R and 2-R, modify the DNA migration as cisplatin, cationic platinacycle 5-R was able to inhibit topoisomerase I-promoted DNA supercoiling, and Pt(ii) coordination compound 7-R turned out to be the most potent inhibitor of cathepsin B. Induction of G-1 phase ( 2-R and 5-R ), and S and G-2 phases (6-R) arrests are related to the antiproliferative activity of some representative compounds upon A-549 cells. Induction of apoptosis is also observed for 2-R and 6-R.

  10. Modeling and three dimensional simulation of the neutral dynamics in an air discharge confined in a microcavity. II. Analysis of the wall and geometry effects

    NASA Astrophysics Data System (ADS)

    Eichwald, O.; Bayle, P.; Yousfi, Y.; Jugroot, M.

    1998-11-01

    This article is devoted to the analysis of the confinement effects of the neutral dynamics generated by a short-gap (0.5 mm) discharge inside three different microcavity geometries (cylinder, cube, and bricklike) filled with air at atmospheric pressure (760 Torr) and ambient temperature (293 K). The discharge is modelled by two mathematical functions representing the Joule heating and the momentum transfer between charged and neutral particles. Their spatio-temporal evolution are taken from experimental results with 470 ns for the duration and 13.5 W for the maximum injected power. The neutral gas evolution is described by the classical transport equations and solved by a powerful numerical monotonic upstream-centered scheme for conversion laws. Because of the microcavity dimensions considered, particular care has been used in the analysis of the thermal and hydrodynamics boundary layers which condition the gas-solid interaction in terms of viscous slip effects and thermal exchanges. The results presented show the microcavity geometry effects on the distribution of the initial cylindrical pressure wave as soon as it reaches the lateral walls. They show the specificity of the cube and bricklike microcavities due to the delayed reflections on the corners leading to a more heterogeneous gas behavior than in the case of the cylindrical microcavity. We also discuss the specific gas behaviors near the wall resulting from heat exchange and viscous stress.

  11. Bactericidal Dendritic Polycation Cloaked with Stealth Material via Lipase-Sensitive Intersegment Acquires Neutral Surface Charge without Losing Membrane-Disruptive Activity.

    PubMed

    Xu, Lulu; He, Chen; Hui, Liwei; Xie, Yuntao; Li, Jia-Min; He, Wei-Dong; Yang, Lihua

    2015-12-23

    Net cationicity of membrane-disruptive antimicrobials is necessary for their activity but may elicit immune attack when administered intravenously. By cloaking a dendritic polycation (G2) with poly(caprolactone-b-ethylene glycol) (PCL-b-PEG), we obtain a nanoparticle antimicrobial, G2-g-(PCL-b-PEG), which exhibits neutral surface charge but kills >99.9% of inoculated bacterial cells at ≤8 μg/mL. The observed activity may be attributed PCL's responsive degradation by bacterial lipase and the consequent exposure of the membrane-disruptive, bactericidal G2 core. Moreover, G2-g-(PCL-b-PEG) exhibits good colloidal stability in the presence of serum and insignificant hemolytic toxicity even at ≥2048 μg/mL. suggesting good blood compatibility required for intravenous administration.

  12. Bactericidal Dendritic Polycation Cloaked with Stealth Material via Lipase-Sensitive Intersegment Acquires Neutral Surface Charge without Losing Membrane-Disruptive Activity.

    PubMed

    Xu, Lulu; He, Chen; Hui, Liwei; Xie, Yuntao; Li, Jia-Min; He, Wei-Dong; Yang, Lihua

    2015-12-23

    Net cationicity of membrane-disruptive antimicrobials is necessary for their activity but may elicit immune attack when administered intravenously. By cloaking a dendritic polycation (G2) with poly(caprolactone-b-ethylene glycol) (PCL-b-PEG), we obtain a nanoparticle antimicrobial, G2-g-(PCL-b-PEG), which exhibits neutral surface charge but kills >99.9% of inoculated bacterial cells at ≤8 μg/mL. The observed activity may be attributed PCL's responsive degradation by bacterial lipase and the consequent exposure of the membrane-disruptive, bactericidal G2 core. Moreover, G2-g-(PCL-b-PEG) exhibits good colloidal stability in the presence of serum and insignificant hemolytic toxicity even at ≥2048 μg/mL. suggesting good blood compatibility required for intravenous administration. PMID:26632646

  13. Techniques of Neutralization and Persistent Sexual Abuse by Clergy: A Content Analysis of Priest Personnel Files From the Archdiocese of Milwaukee.

    PubMed

    Spraitz, Jason D; Bowen, Kendra N

    2016-09-01

    The sexual abuse problem in the Catholic Church has received considerable attention by the media in recent years and growing attention from empirical researchers. Despite this growth, there is a lack of theoretical research that uses neutralization techniques to examine clergy offending. Using Sykes and Matza's theory, this study examines the techniques of neutralization used by accused priests in the Archdiocese of Milwaukee. Priests' personnel files, which were made publicly available by the Archbishop of Milwaukee in July 2013, were analyzed retrospectively through a qualitative content analysis of all direct statements and correspondences from the accused. The findings indicate that many priests denied responsibility or injury in an effort to justify their sexually abusive behaviors, but that no discernible patterns of technique use emerged. The need for continued research using recently released personnel files from other dioceses is also discussed.

  14. An analysis of post-vocalic /s-ʃ/ neutralization in Augsburg German: evidence for a gradient sound change

    PubMed Central

    Bukmaier, Véronique; Harrington, Jonathan; Kleber, Felicitas

    2014-01-01

    The study is concerned with a sound change in progress by which a post-vocalic, pre-consonantal /s-ʃ/ contrast in the standard variety of German (SG) in words such as west/wäscht (/vɛst/~/vɛʃt/, west/washes) is influencing the Augsburg German (AG) variety in which they have been hitherto neutralized as /veʃt/. Two of the main issues to be considered are whether the change is necessarily categorical; and the extent to which the change affects both speech production and perception equally. For the production experiment, younger and older AG and SG speakers merged syllables of hypothetical town names to create a blend at the potential neutralization site. These results showed a trend for a progressively greater /s-ʃ/ differentiation in the order older AG, younger AG, and SG speakers. For the perception experiment, forced-choice responses were obtained from the same subjects who had participated in the production experiment to a 16-step /s-ʃ/ continuum that was embedded into two contexts: /mIst-mIʃt/ in which /s-ʃ/ are neutralized in AG and /və'mIsə/-/və'mIʃə/ in which they are not. The results from both experiments are indicative of a sound change in progress such that the neutralization is being undone under the influence of SG, but in such a way that there is a gradual shift between categories. The closer approximation of the groups on perception suggests that the sound change may be more advanced on this modality than in production. Overall, the findings are consistent with the idea that phonological contrasts are experience-based, i.e., a continuous function of the extent to which a subject is exposed to, and makes use of, the distinction and are thus compatible with exemplar models of speech. PMID:25132828

  15. Characterization of a plant-produced recombinant human secretory IgA with broad neutralizing activity against HIV

    PubMed Central

    Paul, Matthew; Reljic, Rajko; Klein, Katja; Drake, Pascal MW; van Dolleweerd, Craig; Pabst, Martin; Windwarder, Markus; Arcalis, Elsa; Stoger, Eva; Altmann, Friedrich; Cosgrove, Catherine; Bartolf, Angela; Baden, Susan; Ma, Julian K-C

    2014-01-01

    Recombinant Secretory IgA (SIgA) complexes have the potential to improve antibody-based passive immunotherapeutic approaches to combat many mucosal pathogens. In this report, we describe the expression, purification and characterization of a human SIgA format of the broadly neutralizing anti-HIV monoclonal antibody (mAb) 2G12, using both transgenic tobacco plants and transient expression in Nicotiana benthamiana as expression hosts (P2G12 SIgA). The resulting heterodecameric complexes accumulated in intracellular compartments in leaf tissue, including the vacuole. SIgA complexes could not be detected in the apoplast. Maximum yields of antibody were 15.2 μg/g leaf fresh mass (LFM) in transgenic tobacco and 25 μg/g LFM after transient expression, and assembly of SIgA complexes was superior in transgenic tobacco. Protein L purified antibody specifically bound HIV gp140 and neutralised tier 2 and tier 3 HIV isolates. Glycoanalysis revealed predominantly high mannose structures present on most N-glycosylation sites, with limited evidence for complex glycosylation or processing to paucimannosidic forms. O-glycan structures were not identified. Functionally, P2G12 SIgA, but not IgG, effectively aggregated HIV virions. Binding of P2G12 SIgA was observed to CD209 / DC-SIGN, but not to CD89 / FcalphaR on a monocyte cell line. Furthermore, P2G12 SIgA demonstrated enhanced stability in mucosal secretions in comparison to P2G12 IgG mAb. PMID:25484063

  16. Improvement of activated carbons as oxygen reduction catalysts in neutral solutions by ammonia gas treatment and their performance in microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Watson, Valerie J.; Nieto Delgado, Cesar; Logan, Bruce E.

    2013-11-01

    Commercially available activated carbon (AC) powders from different precursor materials (peat, coconut shell, coal, and hardwood) were treated with ammonia gas at 700 °C to improve their performance as oxygen reduction catalysts in neutral pH solutions used in microbial fuel cells (MFCs). The ammonia treated ACs exhibited better catalytic performance in rotating ring-disk electrode tests than their untreated precursors, with the bituminous based AC most improved, with an onset potential of Eonset = 0.12 V (untreated, Eonset = 0.08 V) and n = 3.9 electrons transferred in oxygen reduction (untreated, n = 3.6), and the hardwood based AC (treated, Eonset = 0.03 V, n = 3.3; untreated, Eonset = -0.04 V, n = 3.0). Ammonia treatment decreased oxygen content by 29-58%, increased nitrogen content to 1.8 atomic %, and increased the basicity of the bituminous, peat, and hardwood ACs. The treated coal based AC cathodes had higher maximum power densities in MFCs (2450 ± 40 mW m-2) than the other AC cathodes or a Pt/C cathode (2100 ± 1 mW m-2). These results show that reduced oxygen abundance and increased nitrogen functionalities on the AC surface can increase catalytic performance for oxygen reduction in neutral media.

  17. Neutralizing antibodies against West Nile virus identified directly from human B cells by single-cell analysis and next generation sequencing.

    PubMed

    Tsioris, Konstantinos; Gupta, Namita T; Ogunniyi, Adebola O; Zimnisky, Ross M; Qian, Feng; Yao, Yi; Wang, Xiaomei; Stern, Joel N H; Chari, Raj; Briggs, Adrian W; Clouser, Christopher R; Vigneault, Francois; Church, George M; Garcia, Melissa N; Murray, Kristy O; Montgomery, Ruth R; Kleinstein, Steven H; Love, J Christopher

    2015-12-01

    West Nile virus (WNV) infection is an emerging mosquito-borne disease that can lead to severe neurological illness and currently has no available treatment or vaccine. Using microengraving, an integrated single-cell analysis method, we analyzed a cohort of subjects infected with WNV - recently infected and post-convalescent subjects - and efficiently identified four novel WNV neutralizing antibodies. We also assessed the humoral response to WNV on a single-cell and repertoire level by integrating next generation sequencing (NGS) into our analysis. The results from single-cell analysis indicate persistence of WNV-specific memory B cells and antibody-secreting cells in post-convalescent subjects. These cells exhibited class-switched antibody isotypes. Furthermore, the results suggest that the antibody response itself does not predict the clinical severity of the disease (asymptomatic or symptomatic). Using the nucleotide coding sequences for WNV-specific antibodies derived from single cells, we revealed the ontogeny of expanded WNV-specific clones in the repertoires of recently infected subjects through NGS and bioinformatic analysis. This analysis also indicated that the humoral response to WNV did not depend on an anamnestic response, due to an unlikely previous exposure to the virus. The innovative and integrative approach presented here to analyze the evolution of neutralizing antibodies from natural infection on a single-cell and repertoire level can also be applied to vaccine studies, and could potentially aid the development of therapeutic antibodies and our basic understanding of other infectious diseases. PMID:26481611

  18. Quantitative analysis of serum neutralization of human immunodeficiency virus type 1 from subtypes A, B, C, D, E, F, and I: lack of direct correlation between neutralization serotypes and genetic subtypes and evidence for prevalent serum-dependent infectivity enhancement.

    PubMed Central

    Kostrikis, L G; Cao, Y; Ngai, H; Moore, J P; Ho, D D

    1996-01-01

    Human immunodeficiency virus type 1 (HIV-1) M group strains have been assigned to date to nine distinct genetic subtypes, designated A through I, according to phylogenetic analyses of nucleotide sequences of their env or gag genes. Whether there is any relationship between phylogenetic subtypes and the neutralization serotypes is not clear, yet defining the nature of any such relationship by mathematical means would be of major importance for the development of globally effective HIV-1 vaccines. We have therefore developed a quantitative method to analyze serum neutralization of HIV-1 isolates and to identify HIV-1 neutralization serotypes. This method involves calculations of the neutralization index, N(i), a newly defined parameter derived from plots generated from in vitro neutralization assays, calculations of pairwise serum-virus vector distances, and cluster analyses. We have applied this approach to analyze three independent neutralization matrices involving primary HIV-1 strains and sera from genetic subtypes A, B, C, D, E, F, and I. Detailed serum and HIV-1 isolate cluster analyses have shown that in general, the identified neutralization serotypes do not directly correlate with HIV-1 genetic subtypes. These results suggest that neutralization serotypes do not during natural HIV-1 infection are not governed by antibodies directed against simple epitopes within gp120 monomers. A significant proportion (28%) of 1,213 combinations of sera and HIV-1 isolates caused serum-dependent infectivity enhancement [negative N(i) values] rather than neutralization. We also noted that negative N(i) values tended to correlate better with certain HIV-1 isolates rather than with HIV-1-positive sera. Syncytium-inducing variants of HIV-1 were slightly more likely than non-syncytium-inducing variants to undergo serum-dependent infectivity enhancement, although the latter variants could clearly be susceptible to enhancement. PMID:8523557

  19. A systematic analysis of a broadly neutralizing antibody AR3C epitopes on Hepatitis C virus E2 envelope glycoprotein and their cross-reactivity

    PubMed Central

    2015-01-01

    Background Hepatitis C virus (HCV) belongs to Flaviviridae family of viruses. HCV represents a major challenge to public health since its estimated global prevalence is 2.8% of the world's human population. The design and development of HCV vaccine has been hampered by rapid evolution of viral quasispecies resulting in antibody escape variants. HCV envelope glycoprotein E1 and E2 that mediate fusion and entry of the virus into host cells are primary targets of the host immune responses. Results Structural characterization of E2 core protein and a broadly neutralizing antibody AR3C together with E1E2 sequence information enabled the analysis of B-cell epitope variability. The E2 binding site by AR3C and its surrounding area were identified from the crystal structure of E2c-AR3C complex. We clustered HCV strains using the concept of "discontinuous motif/peptide" and classified B-cell epitopes based on their similarity. Conclusions The assessment of antibody neutralizing coverage provides insights into potential cross-reactivity of the AR3C neutralizing antibody across a large number of HCV variants. PMID:26681161

  20. Neutral buoyancy test evaluation of hardware and extravehicular activity procedures for on-orbit assembly of a 14 meter precision reflector

    NASA Technical Reports Server (NTRS)

    Heard, Walter L., Jr.; Lake, Mark S.

    1993-01-01

    A procedure that enables astronauts in extravehicular activity (EVA) to perform efficient on-orbit assembly of large paraboloidal precision reflectors is presented. The procedure and associated hardware are verified in simulated Og (neutral buoyancy) assembly tests of a 14 m diameter precision reflector mockup. The test article represents a precision reflector having a reflective surface which is segmented into 37 individual panels. The panels are supported on a doubly curved tetrahedral truss consisting of 315 struts. The entire truss and seven reflector panels were assembled in three hours and seven minutes by two pressure-suited test subjects. The average time to attach a panel was two minutes and three seconds. These efficient assembly times were achieved because all hardware and assembly procedures were designed to be compatible with EVA assembly capabilities.

  1. Analysis of the saturation phenomena of the neutralization rate of positively charged 218Po in water vapor.

    PubMed

    Tan, Yanliang; Xiao, Detao; Shan, Jian; Zhou, Qingzhi; Qu, Jingnian

    2014-09-01

    Generally, 88% of the freshly generated 218Po ions decayed from 222Rn are positively charged. These positive ions become neutralized by recombination with negative ions, and the main source of the negative ions is the OH- ions formed by radiolysis of water vapor. However, the neutralization rate of positively charged 218Po versus the square root of the concentration of H2O will be a constant when the concentration of H2O is sufficiently high. Since the electron affinity of the hydroxyl radical formed by water vapor is high, the authors propose that the hydroxyl radical can grab an electron to become OH-. Because the average period of collision with other positively charged ions and the average life of the OH- are much longer than those of the electron, the average concentration of negative ions will grow when the water vapor concentration increases. The authors obtained a model to describe the growth of OH- ions. From this model, it was found that the maximum value of the OH- ion concentration is limited by the square root of the radon concentration. If the radon concentration is invariant, the OH- ion concentration should be approximately a constant when the water vapor concentration is higher than a certain value. The phenomenon that the neutralization rate of positively charged 218Po versus the square root of the water vapor concentration will be saturated when the water vapor concentration is sufficiently high can be explained by this mechanism. This mechanism can be used also to explain the phenomenon that the detection efficiency of a radon monitor based on the electrostatic collection method seems to be constant when the water vapor concentration is high.

  2. Analysis of the saturation phenomena of the neutralization rate of positively charged 218Po in water vapor.

    PubMed

    Tan, Yanliang; Xiao, Detao; Shan, Jian; Zhou, Qingzhi; Qu, Jingnian

    2014-09-01

    Generally, 88% of the freshly generated 218Po ions decayed from 222Rn are positively charged. These positive ions become neutralized by recombination with negative ions, and the main source of the negative ions is the OH- ions formed by radiolysis of water vapor. However, the neutralization rate of positively charged 218Po versus the square root of the concentration of H2O will be a constant when the concentration of H2O is sufficiently high. Since the electron affinity of the hydroxyl radical formed by water vapor is high, the authors propose that the hydroxyl radical can grab an electron to become OH-. Because the average period of collision with other positively charged ions and the average life of the OH- are much longer than those of the electron, the average concentration of negative ions will grow when the water vapor concentration increases. The authors obtained a model to describe the growth of OH- ions. From this model, it was found that the maximum value of the OH- ion concentration is limited by the square root of the radon concentration. If the radon concentration is invariant, the OH- ion concentration should be approximately a constant when the water vapor concentration is higher than a certain value. The phenomenon that the neutralization rate of positively charged 218Po versus the square root of the water vapor concentration will be saturated when the water vapor concentration is sufficiently high can be explained by this mechanism. This mechanism can be used also to explain the phenomenon that the detection efficiency of a radon monitor based on the electrostatic collection method seems to be constant when the water vapor concentration is high. PMID:25068963

  3. Activation of neutral-sphingomyelinase, MAPKs, and p75 NTR-mediating caffeic acid phenethyl ester–induced apoptosis in C6 glioma cells

    PubMed Central

    2014-01-01

    Background Caffeic acid phenethyl ester (CAPE), a component of propolis, is reported to possess anti-inflammatory, anti-bacterial, anti-viral, and anti-tumor activities. Previously, our laboratory demonstrated the in vitro and in vivo bioactivity of CAPE and addressed the role of p53 and the p38 mitogen-activated protein kinase (MAPK) pathway in regulating CAPE-induced apoptosis in C6 glioma cells. Results C6 cancer cell lines were exposed to doses of CAPE; DNA fragmentation and MAPKs and NGF/P75NTR levels were then determined. SMase activity and ceramide content measurement as well as western blotting analyses were performed to clarify molecular changes. The present study showed that CAPE activated neutral sphingomyelinase (N-SMase), which led to the ceramide-mediated activation of MAPKs, including extracellular signal-regulated kinase (ERK), Jun N-terminus kinase (JNK), and p38 MAPK. In addition, CAPE increased the expression of nerve growth factor (NGF) and p75 neurotrophin receptor (p75NTR). The addition of an N-SMase inhibitor, GW4869, established that NGF/p75NTR was the downstream target of N-SMase/ceramide. Pretreatment with MAPK inhibitors demonstrated that MEK/ERK and JNK acted upstream and downstream, respectively, of NGF/p75NTR. Additionally, CAPE-induced caspase 3 activation and poly [ADP-ribose] polymerase cleavage were reduced by pretreatment with MAPK inhibitors, a p75NTR peptide antagonist, or GW4869. Conclusions Taken together, N-SMase activation played a pivotal role in CAPE-induced apoptosis by activation of the p38 MAPK pathway and NGF/p75NTR may explain a new role of CAPE induced apoptosis in C6 glioma. PMID:24997497

  4. Some contributions to the thin-layer chromatographic analysis of complex natural phospholipid and neutral lipid mixtures.

    PubMed

    Helmy, F M; Hack, M H

    1986-01-10

    We have isolated a minor phosphatidyl ethanolamine component from pancreas and a minor phosphatidyl choline component from retina, which were revealed by their separate thin-layer chromatographic properties on silica gel and aluminum oxide sheets, respectively. We have described in some detail a number of modifications in thin-layer chromatography methodology which enhances the opportunity for assessing the glycerophospholipid and neutral lipid composition of tissues, as attested by a diverse set of examples, and have pointed out some of the associated technical problems.

  5. In vivo selection for the enhancement of Thermotoga maritima exopolygalacturonase activity at neutral pH and low temperature.

    PubMed

    Drone, Jullien; Dion, Michel; Tellier, Charles; Rabiller, Claude

    2007-01-01

    The aim of this study was to develop an Escherichia coli-based metabolic selection system for the uncovering of new oligogalacturonate-active enzymes. Based on the expression of the specific permease TogMNAB, this system enabled the entry of oligogalacturonates into the cytoplasm of E. coli thus providing a modified strain usable for this purpose. This tool was used for the metabolic selection of Thermotoga maritima exopolygalacturonase (TmGalU) mutants enabling the uptake of sodium trigalacturonate as the sole carbon source by the bacterium. In only one round of error-prone PCR and selection, mutants of TmGalU with a 4-fold increased turnover at pH 7.0 and 2-fold more active at 37 degrees C than wild-type enzyme were isolated. These results show the versatility of this strain for the evolution of oligogalacturonate-active enzymes.

  6. A Label as a Hidden Persuader: Chemists' Neutralization Concept.

    ERIC Educational Resources Information Center

    Schmidt, Hans-Jurgen

    1991-01-01

    Identifies and describes the problems students have with the concept of neutralization. Analysis of over 7,500 students' answers to test questions over neutralization showed that many students understand the concept in its original meaning. Students assumed that in any neutralization reaction a neutral solution is formed, even if a weak acid or…

  7. Fowlicidin-3 is an alpha-helical cationic host defense peptide with potent antibacterial and lipopolysaccharide-neutralizing activities.

    PubMed

    Bommineni, Yugendar R; Dai, Huaien; Gong, Yu-Xi; Soulages, Jose L; Fernando, Samodha C; Desilva, Udaya; Prakash, Om; Zhang, Guolong

    2007-01-01

    Cathelicidins are an important family of cationic host defense peptides in vertebrates with both antimicrobial and immunomodulatory activities. Fowlicidin-1 and fowlicidin-2 are two newly identified chicken cathelicidins with potent antibacterial activities. Here we report structural and functional characterization of the putatively mature form of the third chicken cathelicidin, fowlicidin-3, for exploration of its therapeutic potential. NMR spectroscopy revealed that fowlicidin-3 comprises 27 amino-acid residues and adopts a predominantly alpha-helical structure extending from residue 9 to 25 with a slight kink induced by a glycine at position 17. It is highly potent against a broad range of Gram-negative and Gram-positive bacteria in vitro, including antibiotic-resistant strains, with minimum inhibitory concentrations in the range 1-2 microM. It kills bacteria quickly, permeabilizing cytoplasmic membranes immediately on coming into contact with them. Unlike many other host defense peptides with antimicrobial activities that are diminished by serum or salt, fowlicidin-3 retains bacteria-killing activities in the presence of 50% serum or physiological concentrations of salt. Furthermore, it is capable of suppressing lipopolysaccharide-induced expression of proinflammatory genes in mouse macrophage RAW264.7 cells, with nearly complete blockage at 10 microM. Fowlicidin-3 appears to be an excellent candidate for future development as a novel antimicrobial and antisepsis agent, particularly against antibiotic-resistant pathogens.

  8. Glucose Administration Enhances fMRI Brain Activation and Connectivity Related to Episodic Memory Encoding for Neutral and Emotional Stimuli

    ERIC Educational Resources Information Center

    Parent, Marise B.; Krebs-Kraft, Desiree L.; Ryan, John P.; Wilson, Jennifer S.; Harenski, Carla; Hamann, Stephan

    2011-01-01

    Glucose enhances memory in a variety of species. In humans, glucose administration enhances episodic memory encoding, although little is known regarding the neural mechanisms underlying these effects. Here we examined whether elevating blood glucose would enhance functional MRI (fMRI) activation and connectivity in brain regions associated with…

  9. Fast targeted analysis of 132 acidic and neutral drugs and poisons in whole blood using LC-MS/MS.

    PubMed

    Di Rago, Matthew; Saar, Eva; Rodda, Luke N; Turfus, Sophie; Kotsos, Alex; Gerostamoulos, Dimitri; Drummer, Olaf H

    2014-10-01

    The aim of this study was to develop an LC-MS/MS based screening technique that covers a broad range of acidic and neutral drugs and poisons by combining a small sample volume and efficient extraction technique with simple automated data processing. After protein precipitation of 100μL of whole blood, 132 common acidic and neutral drugs and poisons including non-steroidal anti-inflammatory drugs, barbiturates, anticonvulsants, antidiabetics, muscle relaxants, diuretics and superwarfarin rodenticides (47 quantitated, 85 reported as detected) were separated using a Shimadzu Prominence HPLC system with a C18 separation column (Kinetex XB-C18, 4.6mm×150mm, 5μm), using gradient elution with a mobile phase of 25mM ammonium acetate buffer (pH 7.5)/acetonitrile. The drugs were detected using an ABSciex(®) API 2000 LC-MS/MS system (ESI+ and -, MRM mode, two transitions per analyte). The method was fully validated in accordance with international guidelines. Quantification data obtained using one-point calibration compared favorably to that using multiple calibrants. The presented LC-MS/MS assay has proven to be applicable for determination of the analytes in blood. The fast and reliable extraction method combined with automated processing gives the opportunity for high throughput and fast turnaround times for forensic and clinical toxicology.

  10. Neutral particle analyzer diagnostics on the TCV tokamak

    SciTech Connect

    Karpushov, Alexander N.; Duval, Basil P.; Schlatter, Christian; Afanasyev, Valery I.; Chernyshev, Fedor V.

    2006-03-15

    Experimental apparatus and data analysis techniques used in neutral particle analyzer (NPA) diagnostics on the Tokamak a Configuration Variable (TCV) are described. Two NPAs are used on TCV to measure the energy spectrum of neutral particle fluxes from the plasma. The 'five-channel energy analyzer of atomic particles' used in double electrical analysis mode with fast voltage sweeping detect particles without atomic mass discrimination in the energy range of 0.6-8.0 keV with a time resolution of 0.5-2.0 ms and an energy resolution of 7%-20%. The 28-channel ''compact neutral particle analyzer'' (CNPA) is an EIIB spectrometer with mass and energy separations designed for medium sized fusion machines featuring a carbon neutral stripping foil, a permanent magnet for dispersion, and channel-electron multiplier detectors. The CNPA simultaneously detects two mass species [hydrogen (H) and deuterium (D) or D and helium (He)] in the 0.5-50 keV energy range with a resolution of 60%-10% and a time resolution of 0.5-4.0 ms. The CNPA views the plasma across the path of the diagnostic neutral beam and can perform active charge-exchange NPA measurement. Data analysis procedures and numerical algorithms developed for NPA measurement are routinely used on TCV to obtain information on the plasma ion temperature, ion energy distribution function, plasma isotope ratios, and other plasma characteristics.

  11. Generation and reactivity of putative support systems, Ce-Al neutral binary oxide nanoclusters: CO oxidation and C-H bond activation.

    PubMed

    Wang, Zhe-Chen; Yin, Shi; Bernstein, Elliot R

    2013-11-21

    Both ceria (CeO2) and alumina (Al2O3) are very important catalyst support materials. Neutral binary oxide nanoclusters (NBONCs), CexAlyOz, are generated and detected in the gas phase and their reactivity with carbon monoxide (CO) and butane (C4H10) is studied. The very active species CeAlO4 (●) can react with CO and butane via O atom transfer (OAT) and H atom transfer (HAT), respectively. Other CexAlyOz NBONCs do not show reactivities toward CO and C4H10. The structures, as well as the reactivities, of CexAlyOz NBONCs are studied theoretically employing density functional theory (DFT) calculations. The ground state CeAlO4 (●) NBONC possesses a kite-shaped structure with an OtCeObObAlOt configuration (Ot, terminal oxygen; Ob, bridging oxygen). An unpaired electron is localized on the Ot atom of the AlOt moiety rather than the CeOt moiety: this Ot centered radical moiety plays a very important role for the reactivity of the CeAlO4 (●) NBONC. The reactivities of Ce2O4, CeAlO4 (●), and Al2O4 toward CO are compared, emphasizing the importance of a spin-localized terminal oxygen for these reactions. Intramolecular charge distributions do not appear to play a role in the reactivities of these neutral clusters, but could be important for charged isoelectronic BONCs. DFT studies show that the reaction of CeAlO4 (●) with C4H10 to form the CeAlO4H●C4H9 (●) encounter complex is barrierless. While HAT processes have been previously characterized for cationic and anionic oxide clusters, the reported study is the first observation of a HAT process supported by a ground state neutral oxide cluster. Mechanisms for catalytic oxidation of CO over surfaces of AlxOy∕MmOn or MmOn∕AlxOy materials are proposed consistent with the presented experimental and theoretical results. PMID:24320332

  12. Total body nitrogen analysis. [neutron activation analysis

    NASA Technical Reports Server (NTRS)

    Palmer, H. E.

    1975-01-01

    Studies of two potential in vivo neutron activation methods for determining total and partial body nitrogen in animals and humans are described. A method using the CO-11 in the expired air as a measure of nitrogen content was found to be adequate for small animals such as rats, but inadequate for human measurements due to a slow excretion rate. Studies on the method of measuring the induced N-13 in the body show that with further development, this method should be adequate for measuring muscle mass changes occurring in animals or humans during space flight.

  13. Antitumor x anti-CD3 bifunctional antibodies redirect T-cells activated in vivo with staphylococcal enterotoxin B to neutralize pulmonary metastases.

    PubMed

    Penna, C; Dean, P A; Nelson, H

    1994-05-15

    T-cell antitumor activities are limited by the requirement of two specific major histocompatibility complex restricted steps, T-helper cell activation and cytotoxic T-lymphocyte targeting. The aim of this study was to investigate whether bypassing these major histocompatibility complex restricted steps using nonspecific in vivo activation of T-cells with staphylococcal enterotoxin-B (SE-B) and retargeting with antitumor x anti-CD3 bifunctional antibody (BFA) could provide an effective antitumor response. C3H/HeN mice were injected i.v. with CL-62 melanoma cells, which express the human melanoma antigen p97, and were treated 10 min later with SE-B and/or anti-CD3 (500A2) x anti-p97 (96.5) BFA. Pulmonary metastases were counted 14 days following injections. SE-B alone induced a dose-dependent activation of T-cells as measured by increased interleukin-2 receptor expression and enhanced proliferative responses. SE-B doses greater than 10 micrograms significantly reduced the number of pulmonary metastases versus control (P < 0.01). Combined treatment with SE-B (50 micrograms) and BFA (5 to 50 micrograms) significantly decreased pulmonary metastases compared to treatment with SE-B alone (P < 0.05). Similar reductions in metastases were observed with the F(ab')2 BFA but not with the unconjugated antitumor component of the BFA. Combined treatments with SE-B plus BFA accomplished better tumor neutralization than adoptively transferred in vitro activated splenocytes (4 x 10(7)) retargeted with BFA (5-100 micrograms; P < 0.05). These studies demonstrate that T-cells can be activated in vivo by SE-B and retargeted with small doses of BFA. In this immunocompetent syngeneic pulmonary metastasis model, SE-B plus BFA provided a dramatic antitumor response. PMID:8168104

  14. The chimeric antibody chLpMab-7 targeting human podoplanin suppresses pulmonary metastasis via ADCC and CDC rather than via its neutralizing activity

    PubMed Central

    Ogasawara, Satoshi; Fujii, Yuki; Oki, Hiroharu; Fukayama, Masashi; Nishioka, Yasuhiko; Kaneko, Mika K.

    2015-01-01

    Podoplanin (PDPN/Aggrus/T1α) binds to C-type lectin-like receptor-2 (CLEC-2) and induces platelet aggregation. PDPN is associated with malignant progression, tumor metastasis, and poor prognosis in several types of cancer. Although many anti-human PDPN (hPDPN) monoclonal antibodies (mAbs), such as D2-40 and NZ-1, have been established, these epitopes are limited to the platelet aggregation-stimulating (PLAG) domain (amino acids 29-54) of hPDPN. Recently, we developed a novel mouse anti-hPDPN mAb, LpMab-7, which is more sensitive than D2-40 and NZ-1, using the Cancer-specific mAb (CasMab) method. The epitope of LpMab-7 was shown to be entirely different from that of NZ-1, a neutralizing mAb against the PLAG domain according to an inhibition assay and lectin microarray analysis. In the present study, we produced a mouse-human chimeric anti-hPDPN mAb, chLpMab-7. ChLpMab-7 showed high antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). Furthermore, chLpMab-7 inhibited the growth of hPDPN-expressing tumors in vivo. Although chLpMab-7 recognizes a non-PLAG domain of hPDPN, it suppressed the hematogenous metastasis of hPDPN-expressing tumors. These results indicated that chLpMab-7 suppressed tumor development and hematogenous metastasis in a neutralization-independent manner. In conclusion, hPDPN shows promise as a target in the development of a novel antibody-based therapy. PMID:26416352

  15. Potent neutralization of VEGF biological activities with a fully human antibody Fab fragment directed against VEGF receptor 2

    SciTech Connect

    Miao, H.-Q. . E-mail: hua-quan.miao@imclone.com; Hu, Kun; Jimenez, Xenia; Navarro, Elizabeth; Zhang, Haifan; Lu Dan; Ludwig, Dale L.; Balderes, Paul; Zhu Zhenping . E-mail: zhenping.zhu@imclone.com

    2006-06-23

    Compelling evidence suggest that vascular endothelial growth factor (VEGF) and its receptors, especially receptor 2 (VEGFR2, or kinase insert domain-containing receptor, KDR), play a critical role in angiogenesis under both physiological and pathological conditions, including cancer and angiogenic retinopathies such as age-related macular degeneration (AMD). To this end, inhibition of angiogenesis with antagonists to either VEGF or KDR has yielded significant therapeutic efficacy both in preclinical studies in animal models and in clinical trials in patients with cancer and AMD. We previously reported the identification of a high affinity, fully human anti-KDR antibody fragment, 1121B Fab, through a highly stringent affinity maturation process with a Fab originally isolated from a naive human antibody phage display library. In this study, we demonstrate that 1121B Fab is able to strongly block KDR/VEGF interaction, resulting in potent inhibition of an array of biological activities of VEGF, including activation of the receptor and its signaling pathway, intracellular calcium mobilization, and migration and proliferation of endothelial cells. Taken together, our data lend strong support to the further development of 1121B Fab fragment as an anti-angiogenesis agent in both cancer and angiogenic retinopathies.

  16. Neutral beam monitoring

    DOEpatents

    Fink, Joel H.

    1981-08-18

    Method and apparatus for monitoring characteristics of a high energy neutral beam. A neutral beam is generated by passing accelerated ions through a walled cell containing a low energy neutral gas, such that charge exchange neutralizes the high energy ion beam. The neutral beam is monitored by detecting the current flowing through the cell wall produced by low energy ions which drift to the wall after the charge exchange. By segmenting the wall into radial and longitudinal segments various beam conditions are further identified.

  17. Analysis of active renin heterogeneity.

    PubMed

    Katz, S A; Malvin, R L; Lee, J; Kim, S H; Murray, R D; Opsahl, J A; Abraham, P A

    1991-09-01

    Active renin is a heterogeneous enzyme that can be separated into multiple forms with high-resolution isoelectric focusing. The isoelectric heterogeneity may result from differences in glycosylation between the different forms. In order to determine the relationship between active renin heterogeneity and differences in composition or attachment of oligosaccharides, two separate experiments were performed: (i) Tunicamycin, which interferes with normal glycosylation processing, increased the proportion of relatively basic renin forms secreted into the incubation media by rat renal cortical slices. (ii) Endoglycosidase F, which enzymatically removes carbohydrate from some classes of glycoprotein, similarly increased the proportion of relatively basic forms when incubated with active human recombinant renin. In addition, further studies with inhibitors of human renin activity revealed that the heterogeneous renin forms were similarly inhibited by two separate renin inhibitors. These results are consistent with the hypothesis that renin isoelectric heterogeneity is due in part to differences in carbohydrate moiety attachment and that the heterogeneity of renin does not influence access of direct renin inhibitors to the active site of renin.

  18. Analysis of active renin heterogeneity.

    PubMed

    Katz, S A; Malvin, R L; Lee, J; Kim, S H; Murray, R D; Opsahl, J A; Abraham, P A

    1991-09-01

    Active renin is a heterogeneous enzyme that can be separated into multiple forms with high-resolution isoelectric focusing. The isoelectric heterogeneity may result from differences in glycosylation between the different forms. In order to determine the relationship between active renin heterogeneity and differences in composition or attachment of oligosaccharides, two separate experiments were performed: (i) Tunicamycin, which interferes with normal glycosylation processing, increased the proportion of relatively basic renin forms secreted into the incubation media by rat renal cortical slices. (ii) Endoglycosidase F, which enzymatically removes carbohydrate from some classes of glycoprotein, similarly increased the proportion of relatively basic forms when incubated with active human recombinant renin. In addition, further studies with inhibitors of human renin activity revealed that the heterogeneous renin forms were similarly inhibited by two separate renin inhibitors. These results are consistent with the hypothesis that renin isoelectric heterogeneity is due in part to differences in carbohydrate moiety attachment and that the heterogeneity of renin does not influence access of direct renin inhibitors to the active site of renin. PMID:1908097

  19. Neutron activation analysis of a penny

    NASA Astrophysics Data System (ADS)

    Stevens, Richard E.

    2000-04-01

    Neutron activation analysis has been used for many years as an analysis tool and as an educational tool to teach students about nuclear properties. This article presents an exercise in the neutron activation analysis of a penny which, due to the simplicity of the resulting gamma-ray spectra, is appropriate for general physics classes. Students express a great deal of interest both in seeing the reactor in use as well as determining the composition of something that is familiar to them.

  20. LACBWR primary shield activation analysis

    SciTech Connect

    Nelson, L.L.; Lahti, G.P.; Johnson, W.J.

    1996-11-01

    Nuclear power plants in the US are required to estimate the costs of decommissioning to ensure that adequate funds are accumulated during the useful life of the plant. A major component of the decommissioning cost is the disposal of radioactive material, including material near the reactor created by neutron activation. An accurate assessment of the residual radioactivity in the reactor`s primary shield is necessary to determine this portion of the decommissioning demolition and disposal cost. This paper describes the efforts used to determine the activation levels remaining in the primary shield of the LaCrosse boiling water reactor (LACBWR), owned and operated by Dairyland Power Cooperative.

  1. A Neutralizing Monoclonal IgG1 Antibody of Platelet-Activating Factor Acetylhydrolase SsE Protects Mice against Lethal Subcutaneous Group A Streptococcus Infection

    PubMed Central

    Liu, Mengyao; Feng, Wenchao; Zhu, Hui

    2015-01-01

    Group A Streptococcus (GAS) can cause life-threatening invasive infections, including necrotizing fasciitis. There are no effective treatments for severe invasive GAS infections. The platelet-activating factor (PAF) acetylhydrolase SsE produced by GAS is required for invasive GAS to evade innate immune responses and to invade soft tissues. This study determined whether the enzymatic activity of SsE is critical for its function in GAS skin invasion and inhibition of neutrophil recruitment and whether SsE is a viable target for immunotherapy for severe invasive GAS infections. An isogenic derivative of M1T1 strain MGAS5005 producing SsE with an S178A substitution (SsES178A), an enzymatically inactive SsE mutant protein, was generated. This strain induced higher levels of neutrophil infiltration and caused smaller lesions than MGAS5005 in subcutaneous infections of mice. This phenotype is similar to that of MGAS5005 sse deletion mutants, indicating that the enzymatic activity of SsE is critical for its function. An anti-SsE IgG1 monoclonal antibody (MAb), 2B11, neutralized the PAF acetylhydrolase activity of SsE. Passive immunization with 2B11 increased neutrophil infiltration, reduced skin invasion, and protected mice against MGAS5005 infection. However, 2B11 did not protect mice when it was administered after MGAS5005 inoculation. MGAS5005 induced vascular effusion at infection sites at early hours after GAS inoculation, suggesting that 2B11 did not always have access to infection sites. Thus, the enzymatic activity of SsE mediates its function, and SsE has the potential to be included in a vaccine but is not a therapeutic target. An effective MAb-based immunotherapy for severe invasive GAS infections may need to target virulence factors that are critical for systemic survival of GAS. PMID:25916987

  2. Functional MRI activation in response to panic-specific, non-panic aversive, and neutral pictures in patients with panic disorder and healthy controls.

    PubMed

    Engel, K R; Obst, K; Bandelow, B; Dechent, P; Gruber, O; Zerr, I; Ulrich, K; Wedekind, D

    2016-09-01

    There is evidence that besides limbic brain structures, prefrontal and insular cortical activations and deactivations are involved in the pathophysiology of panic disorder. This study investigated activation response patterns to stimulation with individually selected panic-specific pictures in patients with panic disorder with agoraphobia (PDA) and healthy control subjects using functional magnetic resonance imaging (fMRI). Structures of interest were the prefrontal, cingulate, and insular cortex, and the amygdalo-hippocampal complex. Nineteen PDA subjects (10 females, 9 males) and 21 healthy matched controls were investigated using a Siemens 3-Tesla scanner. First, PDA subjects gave Self-Assessment Manikin (SAM) ratings on 120 pictures showing characteristic panic/agoraphobia situations, of which 20 pictures with the individually highest SAM ratings were selected. Twenty matched pictures showing aversive but not panic-specific stimuli and 80 neutral pictures from the International Affective Picture System were chosen for each subject as controls. Each picture was shown twice in each of four subsequent blocks. Anxiety and depression ratings were recorded before and after the experiment. Group comparisons revealed a significantly greater activation in PDA patients than control subjects in the insular cortices, left inferior frontal gyrus, dorsomedial prefrontal cortex, the left hippocampal formation, and left caudatum, when PA and N responses were compared. Comparisons for stimulation with unspecific aversive pictures showed activation of similar brain regions in both groups. Results indicate region-specific activations to panic-specific picture stimulation in PDA patients. They also imply dysfunctionality in the processing of interoceptive cues in PDA and the regulation of negative emotionality. Therefore, differences in the functional networks between PDA patients and control subjects should be further investigated. PMID:26585457

  3. Conference on Instrumental Activation Analysis: IAA 89

    NASA Astrophysics Data System (ADS)

    Vobecky, M.; Obrusnik, I.

    1989-05-01

    The proceedings contain 40 abstracts of papers all of which have been incorporated in INIS. The papers were centred on the applications of radioanalytical methods, especially on neutron activation analysis, x ray fluorescence analysis, PIXE analysis and tracer techniques in biology, medicine and metallurgy, measuring instruments including microcomputers, and data processing methods.

  4. RE: Pedagogy--After Neutrality

    ERIC Educational Resources Information Center

    I'Anson, John

    2010-01-01

    Within the UK and in many parts of the world, official accounts of what it is to make sense of religion are framed within a rhetorics of neutrality in which such study is premised upon the possibility of dispassionate engagement and analysis. This paper, which is largely theoretical in scope, explores both the affordances and the costs of such an…

  5. PDX neutral beam reionization losses

    SciTech Connect

    Kugel, H.W.; Dylla, H.F.; Eubank, H.P.; Kozub, T.A.; Moore, R.; Schilling, G.; Stuart, L.D.; Von Halle, A.; Williams, M.D.

    1982-04-01

    Reionization losses for 1.5 MW H /sup 0/ and 2 MW D /sup 0/ neutral beams injected into the PDX tokamak were studied using pressure gauges, phototransistors, thermocouples, surface shielding, and surface sample analysis. Considerable outgassing of conventionally prepared 304 SS ducts occurred during initial injections and gradually decreased with the cumulative absorption of beam power. Reionization power losses are presently about 5% in the ducts and about 12% total for a beamline including the duct. Present duct pressures are attributed primarily to gas from the ion source and neutralizer with much smaller contributions from residual wall desorption. Physical mechanisms for the observed duct outgassing are discussed.

  6. Neutralization of radical toxicity by temperature-dependent modulation of extracellular SOD activity in coral bleaching pathogen Vibrio shiloi and its role as a virulence factor.

    PubMed

    Murali, Malliga Raman; Raja, Subramaniya Bharathi; Devaraj, Sivasitambaram Niranjali

    2010-08-01

    Vibrio shiloi is the first and well-documented bacterium which causes coral bleaching, particularly, during summer, when seawater temperature is between 26 and 31 degrees C. Coral bleaching is the disruption of the symbiotic association between coral hosts and their photosynthetic microalgae zooxanthellae. This is either due to lowered resistance in corals to infection or increased virulence of the bacterium at the higher sea surface temperature. The concentration of the oxygen and resulting oxygen radicals produced by the zooxanthellae during photosynthesis are highly toxic to bacteria, which also assist corals in resisting the infection. Hence, in this study we examined the effect of different temperatures on the activity of a novel extracellular SOD in V. shiloi. We also partially characterized the SOD and clearly confirmed that the extracellular SOD produced by V. shiloi is Mn-SOD type, as it was not inhibited by H2O2 or KCN. Performing chemical susceptibility killing assay, we confirmed that extracellular SOD may act as first line of defense for the bacteria against the reactive oxygen species. Since, increased activity of novel Mn-SOD at higher temperature, leads to the neutralization of radical toxicity and facilitates the survival of V. shiloi. Hence, the extracellular Mn-SOD may be considered as a virulence factor.

  7. Canine distemper virus neutralization activity is low in human serum and it is sensitive to an amino acid substitution in the hemagglutinin protein.

    PubMed

    Zhang, Xinsheng; Wallace, Olivia L; Domi, Arban; Wright, Kevin J; Driscoll, Jonathan; Anzala, Omu; Sanders, Eduard J; Kamali, Anatoli; Karita, Etienne; Allen, Susan; Fast, Pat; Gilmour, Jill; Price, Matt A; Parks, Christopher L

    2015-08-01

    Serum was analyzed from 146 healthy adult volunteers in eastern Africa to evaluate measles virus (MV) and canine distemper virus (CDV) neutralizing antibody (nAb) prevalence and potency. MV plaque reduction neutralization test (PRNT) results indicated that all sera were positive for MV nAbs. Furthermore, the 50% neutralizing dose (ND50) for the majority of sera corresponded to antibody titers induced by MV vaccination. CDV nAbs titers were low and generally were detected in sera with high MV nAb titers. A mutant CDV was generated that was less sensitive to neutralization by human serum. The mutant virus genome had 10 nucleotide substitutions, which coded for single amino acid substitutions in the fusion (F) and hemagglutinin (H) glycoproteins and two substitutions in the large polymerase (L) protein. The H substitution occurred in a conserved region involved in receptor interactions among morbilliviruses, implying that this region is a target for cross-reactive neutralizing antibodies. PMID:25880113

  8. Canine distemper virus neutralization activity is low in human serum and it is sensitive to an amino acid substitution in the hemagglutinin protein.

    PubMed

    Zhang, Xinsheng; Wallace, Olivia L; Domi, Arban; Wright, Kevin J; Driscoll, Jonathan; Anzala, Omu; Sanders, Eduard J; Kamali, Anatoli; Karita, Etienne; Allen, Susan; Fast, Pat; Gilmour, Jill; Price, Matt A; Parks, Christopher L

    2015-08-01

    Serum was analyzed from 146 healthy adult volunteers in eastern Africa to evaluate measles virus (MV) and canine distemper virus (CDV) neutralizing antibody (nAb) prevalence and potency. MV plaque reduction neutralization test (PRNT) results indicated that all sera were positive for MV nAbs. Furthermore, the 50% neutralizing dose (ND50) for the majority of sera corresponded to antibody titers induced by MV vaccination. CDV nAbs titers were low and generally were detected in sera with high MV nAb titers. A mutant CDV was generated that was less sensitive to neutralization by human serum. The mutant virus genome had 10 nucleotide substitutions, which coded for single amino acid substitutions in the fusion (F) and hemagglutinin (H) glycoproteins and two substitutions in the large polymerase (L) protein. The H substitution occurred in a conserved region involved in receptor interactions among morbilliviruses, implying that this region is a target for cross-reactive neutralizing antibodies.

  9. Energetic Neutral Atom Precipitation (ENAP)

    NASA Technical Reports Server (NTRS)

    Tinsley, B. A.

    1988-01-01

    The Energetic Neutral Atom Precipitation experiment is scheduled to be flown on the Atmospheric Laboratory for Applications and Science (ATLAS 1) NASA mission. The objective of this experiment is to measure very faint emissions at nighttime arising from fluxes of energetic neutral atoms in the thermosphere. These energetic atoms have energies ranging up to about 50 keV, and arise from ions of hydrogen, helium, and oxygen trapped in the inner magnetosphere. Some of these ions become neutralized in charge exchange reactions with neutral hydrogen in the hydrogen geocorona that extends through the region. The ions are trapped on magnetic field lines which cross the equatorial plane at 2 to 6 earth radii distance, and they mirror at a range of heights on these field lines, extending down to the thermosphere at 500 km altitude. The ATLAS 1 measurements will not be of the neutral atoms themselves but of the optical emission produced by those on trajectories that intersect the thermosphere. The ENAP measurements are to be made using the Imaging Spectrometric Observatory (ISO) which is being flown on the ATLAS mission primarily for daytime spectral observations, and the ENAP measurements will all be nighttime measurements because of the faintness of the emissions and the relatively low level of magnetic activity expected.

  10. Use of zirconium oxychloride to neutralize HF in the microwave-assisted acid dissolution of ceramic glazes for their chemical analysis by ICP-OES.

    PubMed

    Dondi, M; Fabbri, B; Mingazzini, C

    1998-04-01

    The use of a zirconium compound (ZrOCl(2)) to neutralize HF in the microwave-assisted acid dissolution of ceramic glazes for their chemical analysis was tested. Zr is a strong complexing agent for the fluorine ion and permits the determination of those elements which would form insoluble fluorides. The use of Zr implies strong spectral interferences and a high salt content; however, we found that at least 27 elements can be measured with low detection limits, and satisfactory precision and accuracy. In addition, the use of ZrOCl(2) would permit the complete analysis of a ceramic glaze with a single attack when acid-resistant mineral phases are not present. For potassium determinations in acid matrix, the addition of an ionization buffer was studied in order to increase sensitivity, avoiding ionization interferences and non-linear calibration curves. PMID:18967112

  11. Quantitative Surface Analysis of a Binary Drug Mixture—Suppression Effects in the Detection of Sputtered Ions and Post-Ionized Neutrals

    NASA Astrophysics Data System (ADS)

    Karras, Gabriel; Lockyer, Nicholas P.

    2014-05-01

    A systematic mass spectrometric study of two of the most common analgesic drugs, paracetamol and ibuprofen, is reported. The drugs were studied by means of secondary ion mass spectrometry (SIMS) and secondary neutral mass spectrometry (SNMS) using laser post-ionization (LPI) both in pure samples and in a two-component mixture. Ion suppression within the two-component system observed in SIMS mode is ameliorated using LPI under room temperature analysis. However, suppression effects are apparent in LPI mode on performing the analysis at cryogenic temperatures, which we attribute to changes in the desorption characteristics of sputtered molecules, which influences the subsequent post-ionization efficiency. This suggests different mechanisms of ion suppression in SIMS and LPI modes.

  12. Energetic Neutral Atom Imaging at Low Altitudes from the Swedish Microsatellite Astrid: Images and Spectral Analysis. Paper 1

    NASA Technical Reports Server (NTRS)

    Brandt, Pontus C:son; Barabash, Stas; Norberg, Olle; Lundin, Rickard; Roelof, Rdmond C.; Chase, Christopher J.

    1999-01-01

    Observations of energetic neutral atoms (ENA) in the energy range 26- 52 keV are reported from four occasions during geomagnetically disturbed periods. The data were acquired by the ENA imager flown on the Swedish microsatellite Astrid in a 1000 km circular orbit with 83 deg inclination. The ENA imager separates charged particles from neutrals through an electrostatic deflection system in the energy range between 0.1 and 114 keV. ENA images obtained from vantage points in the polar cap and in the afternoon magnetic local time (MLT) hours looking into the antisunward hemisphere show intense ENA fluxes (approx. 10(exp 4)/sq cm sr s over 26-37 keV) coming from the dusk region and low altitudes (approx. 300 km). The morphology shows no relation to local magnetic field excluding the possibility of charged particle detection. It is concluded that the source of these ENAs are precipitating/mirroring ions from the ring current/trapped radiation interacting with the exobase on auroral L-shells and in the dusk region. The observed ENA fluxes show a relation with Kp and Dst geomagnetic indices. The observed ENA spectrum from a geomagnetic storm on February 8, 1995, is investigated in more detail and compared to the parent ion spectrum obtained by the Defense Meteorological Satellite Project (DMSP) satellite, Fl2, during the same period on L = 6 +/- 2 around dusk. The observed ENA spectral slope is used to derive the parent ion spectral temperature. The derived ion temperatures range is 3.0 - 6.0 keV for H and 4.5 - 8.5 keV for O. The higher of these ion temperatures comes closest in agreement to the extrapolated DMSP spectrum leading us to favor O over H as the species of the detected ENAS. It is shown that the detected ENAs must have been produced at L greater than or equal to 6 to reach the detector without atmospheric attenuation and that the main energy dependence of the ENA spectrum, apart from the parent ion spectrum, is governed by the energy dependence of the charge

  13. Phylogenetic analysis of the main neutralization and hemagglutination determinants of all human adenovirus prototypes as a basis for molecular classification and taxonomy.

    PubMed

    Madisch, Ijad; Harste, Gabi; Pommer, Heidi; Heim, Albert

    2005-12-01

    Human adenoviruses (HAdV) are responsible for a wide spectrum of diseases. The neutralization epsilon determinant (loops 1 and 2) and the hemagglutination gamma determinant are relevant for the taxonomy of HAdV. Precise type identification of HAdV prototypes is crucial for detection of infection chains and epidemiology. epsilon and gamma determinant sequences of all 51 HAdV were generated to propose molecular classification criteria. Phylogenetic analysis of epsilon determinant sequences demonstrated sufficient genetic divergence for molecular classification, with the exception of HAdV-15 and HAdV-29, which also cannot be differentiated by classical cross-neutralization. Precise sequence divergence criteria for typing (<2.5% from loop 2 prototype sequence and <2.4% from loop 1 sequence) were deduced from phylogenetic analysis. These criteria may also facilitate identification of new HAdV prototypes. Fiber knob (gamma determinant) phylogeny indicated a two-step model of species evolution and multiple intraspecies recombination events in the origin of HAdV prototypes. HAdV-29 was identified as a recombination variant of HAdV-15 (epsilon determinant) and a speculative, not-yet-isolated HAdV prototype (gamma determinant). Subanalysis of molecular evolution in hypervariable regions 1 to 6 of the epsilon determinant indicated different selective pressures in subclusters of species HAdV-D. Additionally, gamma determinant phylogenetic analysis demonstrated that HAdV-8 did not cluster with -19 and -37 in spite of their having the same tissue tropism. The phylogeny of HAdV-E4 suggested origination by interspecies recombination between HAdV-B (hexon) and HAdV-C (fiber), as in simian adenovirus 25, indicating additional zoonotic transfer. In conclusion, molecular classification by systematic sequence analysis of immunogenic determinants yields new insights into HAdV phylogeny and evolution.

  14. Phylogenetic Analysis of the Main Neutralization and Hemagglutination Determinants of All Human Adenovirus Prototypes as a Basis for Molecular Classification and Taxonomy

    PubMed Central

    Madisch, Ijad; Harste, Gabi; Pommer, Heidi; Heim, Albert

    2005-01-01

    Human adenoviruses (HAdV) are responsible for a wide spectrum of diseases. The neutralization ɛ determinant (loops 1 and 2) and the hemagglutination γ determinant are relevant for the taxonomy of HAdV. Precise type identification of HAdV prototypes is crucial for detection of infection chains and epidemiology. ɛ and γ determinant sequences of all 51 HAdV were generated to propose molecular classification criteria. Phylogenetic analysis of ɛ determinant sequences demonstrated sufficient genetic divergence for molecular classification, with the exception of HAdV-15 and HAdV-29, which also cannot be differentiated by classical cross-neutralization. Precise sequence divergence criteria for typing (<2.5% from loop 2 prototype sequence and <2.4% from loop 1 sequence) were deduced from phylogenetic analysis. These criteria may also facilitate identification of new HAdV prototypes. Fiber knob (γ determinant) phylogeny indicated a two-step model of species evolution and multiple intraspecies recombination events in the origin of HAdV prototypes. HAdV-29 was identified as a recombination variant of HAdV-15 (ɛ determinant) and a speculative, not-yet-isolated HAdV prototype (γ determinant). Subanalysis of molecular evolution in hypervariable regions 1 to 6 of the ɛ determinant indicated different selective pressures in subclusters of species HAdV-D. Additionally, γ determinant phylogenetic analysis demonstrated that HAdV-8 did not cluster with -19 and -37 in spite of their having the same tissue tropism. The phylogeny of HAdV-E4 suggested origination by interspecies recombination between HAdV-B (hexon) and HAdV-C (fiber), as in simian adenovirus 25, indicating additional zoonotic transfer. In conclusion, molecular classification by systematic sequence analysis of immunogenic determinants yields new insights into HAdV phylogeny and evolution. PMID:16306598

  15. Neutral-current detectors for the Sudbury Neutrino Observatory

    SciTech Connect

    Hime, A.; SNO Collaboration

    1997-09-01

    With its heavy water target, the Sudbury Neutrino Observatory has the unique opportunity to measure both the {sup 8}B flux of electron neutrinos from the Sun and the flux of all active neutrino species independently, thus offering a direct and model-independent test of a neutrino oscillation solution to the solar neutrino problem. The authors report on the physics intent and design of a discrete method of neutral-current detection in the Sudbury neutrino observatory that will utilize ultra-low background {sup 3}He proportional counters dispersed throughout the heavy water volume. Projections of background in all components of the detector are considered in an analysis of the ability to extract the neutral-current signal and the neutral-current to charged-current ratio.

  16. Chiral separation of β-blockers by MEEKC using neutral microemulsion: Analysis of separation mechanism and further elucidation of resolution equation.

    PubMed

    Hu, Shao-Qiang; Lü, Wen-Juan; Ma, Yan-Hua; Hu, Qin; Dong, Li-Jun; Chen, Xing-Guo

    2013-01-01

    Based on the investigation of the effect of microemulsion charge on the chiral separation, a new chiral separation method with MEEKC employing neutral microemulsion was established. The method used a microemulsion containing 3.0% (w/v) neutral surfactant Tween 20 and 0.8% (w/v, 30 mM) dibutyl l-tartrate in 40 mM sodium tetraborate buffer to separate the enantiomers of β-blockers. The effect of major parameters on the chiral separation was investigated. The applied voltage had little effect on the resolution, but the chiral separation could be improved by suppressing the EOF. Nine racemic β-blockers obtained relatively good enantioseparation after appropriate concentrations of tetradecyl trimethyl ammonium bromide were added into the microemulsion to suppress the EOF. These results were explained based on the analysis of the separation mechanism of the method and deduced separation equations. The resolution equation of the method was further elucidated. It was found that the fourth term in the resolution equation, an additional term compared to the conventional resolution equation for column chromatography, represents the ratio of the relative movement distance between the analyte and microemulsion droplets relative to the effective capillary length. It can be regarded as a correction for the effective capillary length. These findings are significant for the development of the theory of MEEKC and the development of new chiral MEEKC method.

  17. On neutral plasma oscillations

    SciTech Connect

    Shadwick, B.A.; Morrison, P.J.

    1993-06-01

    We examine the conditions for the existence of spectrally stable neutral modes in a Vlasov-Poisson plasma and show that for stable equilibria of systems that have unbounded spatial domain, the only possible neutral modes are those with phase velocities that correspond to stationary inflection points of the equilibrium distribution function. It is seen that these neutral modes can possess positive or negative free energy.

  18. ALEX neutral beam probe

    SciTech Connect

    Pourrezaei, K.

    1982-01-01

    A neutral beam probe capable of measuring plasma space potential in a fully 3-dimensional magnetic field geometry has been developed. This neutral beam was successfully used to measure an arc target plasma contained within the ALEX baseball magnetic coil. A computer simulation of the experiment was performed to refine the experimental design and to develop a numerical model for scaling the ALEX neutral beam probe to other cases of fully 3-dimensional magnetic field. Based on this scaling a 30 to 50 keV neutral cesium beam probe capable of measuring space potential in the thermal barrier region of TMX Upgrade was designed.

  19. Search for neutral leptons

    SciTech Connect

    Perl, M.L.

    1984-12-01

    At present we know of three kinds of neutral leptons: the electron neutrino, the muon neutrino, and the tau neutrino. This paper reviews the search for additional neutral leptons. The method and significance of a search depends upon the model used for the neutral lepton being sought. Some models for the properties and decay modes of proposed neutral leptons are described. Past and present searches are reviewed. The limits obtained by some completed searches are given, and the methods of searches in progress are described. Future searches are discussed. 41 references.

  20. Neutralization of the oedematogenic activity of Bothrops jararaca venom on the mouse paw by an antibothropic fraction isolated from opossum (Didelphis marsupialis) serum.

    PubMed

    Perales, J; Amorim, C Z; Rocha, S L; Domont, G B; Moussatché, H

    1992-11-01

    The pharmacological modulation of mice paw oedema produced by Bothrops jararaca venom (BJV) has been studied. Intraplantar injection of BJV (1-30 micrograms/paw) produced a dose- and time-related oedema, which was maximal 30 min after injection, reduced gradually thereafter and disappeared over 48 h. BJV heated at 100 degrees C for 5 or 15 min blocked local hemorrhage and caused partial inhibition of its oedematogenic activity. The BJV oedema was not inhibited by the anti-histamine meclizine, the inhibitor of histamine and serotonin, cyproheptadine, PAF-acether antagonist WEB 2170 or by the anti-leukotrienes C4/D4, LY 171883. Dexamethasone, aspirin, indomethacin, and the dual cyclooxygenase and lipoxygenase inhibitor BW 755C inhibited BJV-induced oedema indicating that arachidonic acid metabolism products via the cyclooxygenase pathway participate in its genesis and/or maintenance. The antibothropic fraction (ABF) (25-200 micrograms/paw) isolated from Didelphis marsupialis serum neutralized the oedema induced by the venom with and without heating, the hemorrhage induced by BJV and partially blocked the oedema induced by bradykinin and by cellulose sulphate. The oedema produced by histamine, serotonin, PAF-acether or leukotriene C4 was not inhibited. PMID:1295374

  1. A microplate assay for the coupled transglycosylase-transpeptidase activity of the penicillin binding proteins; a vancomycin-neutralizing tripeptide combination prevents penicillin inhibition of peptidoglycan synthesis.

    PubMed

    Kumar, Vidya P; Basavannacharya, Chandrakala; de Sousa, Sunita M

    2014-07-18

    A microplate, scintillation proximity assay to measure the coupled transglycosylase-transpeptidase activity of the penicillin binding proteins in Escherichia coli membranes was developed. Membranes were incubated with the two peptidoglycan sugar precursors UDP-N-acetyl muramylpentapeptide (UDP-MurNAc(pp)) and UDP-[(3)H]N-acetylglucosamine in the presence of 40 μM vancomycin to allow in situ accumulation of lipid II. In a second step, vancomycin inhibition was relieved by addition of a tripeptide (Lys-D-ala-D-ala) or UDP-MurNAc(pp), resulting in conversion of lipid II to cross-linked peptidoglycan. Inhibitors of the transglycosylase or transpeptidase were added at step 2. Moenomycin, a transglycosylase inhibitor, had an IC50 of 8 nM. Vancomycin and nisin also inhibited the assay. Surprisingly, the transpeptidase inhibitors penicillin and ampicillin showed no inhibition. In a pathway assay of peptidoglycan synthesis, starting from the UDP linked sugar precursors, inhibition by penicillin was reversed by a 'neutral' combination of vancomycin plus tripeptide, suggesting an interaction thus far unreported. PMID:24944023

  2. A microplate assay for the coupled transglycosylase-transpeptidase activity of the penicillin binding proteins; a vancomycin-neutralizing tripeptide combination prevents penicillin inhibition of peptidoglycan synthesis.

    PubMed

    Kumar, Vidya P; Basavannacharya, Chandrakala; de Sousa, Sunita M

    2014-07-18

    A microplate, scintillation proximity assay to measure the coupled transglycosylase-transpeptidase activity of the penicillin binding proteins in Escherichia coli membranes was developed. Membranes were incubated with the two peptidoglycan sugar precursors UDP-N-acetyl muramylpentapeptide (UDP-MurNAc(pp)) and UDP-[(3)H]N-acetylglucosamine in the presence of 40 μM vancomycin to allow in situ accumulation of lipid II. In a second step, vancomycin inhibition was relieved by addition of a tripeptide (Lys-D-ala-D-ala) or UDP-MurNAc(pp), resulting in conversion of lipid II to cross-linked peptidoglycan. Inhibitors of the transglycosylase or transpeptidase were added at step 2. Moenomycin, a transglycosylase inhibitor, had an IC50 of 8 nM. Vancomycin and nisin also inhibited the assay. Surprisingly, the transpeptidase inhibitors penicillin and ampicillin showed no inhibition. In a pathway assay of peptidoglycan synthesis, starting from the UDP linked sugar precursors, inhibition by penicillin was reversed by a 'neutral' combination of vancomycin plus tripeptide, suggesting an interaction thus far unreported.

  3. Production of monoclonal antibodies capable of neutralizing dermonecrotic activity of Loxosceles intermedia spider venom and their use in a specific immunometric assay.

    PubMed

    Alvarenga, Larissa M; Martins, Maria S; Moura, Juliana F; Kalapothakis, Evanguedes; Oliveira, Júlio C; Mangili, Oldemir C; Granier, Claude; Chávez-Olórtegui, Carlos

    2003-12-01

    We have produced 13 mAbs for Loxosceles intermedia crude venom. Twelve were reactive against proteins of 32-35 kDa and one of these Li mAb(7) showed high neutralizing potency for the dermonecrotic activity of L. intermedia venom. This Li mAb(7) showed no cross-reactivity, with Loxosceles laeta (Brazil), L. laeta (Perú) and Loxosceles gaucho venoms. The mAbs were produced by immunization with the crude venom and screened by enzyme-linked immunosorbent assay (ELISA) using L. intermedia whole venom or dermonecrotic fraction (DNF) as antigens coated onto microtitre plates. A sensitive two-site immunometric assay was designed and shown to be useful for identifying and quantifying DNF from L. intermedia in biological samples. The Li mAb(7) coated onto microtitre plates and hyperimmune horse anti-L. intermedia IgGs prepared by immunoaffinity chromatography and conjugated to horseradish peroxidase were used to set up a sandwich-type ELISA. Measurable absorbance signals were obtained with 0.2 ng of L. intermedia crude venom per assay.

  4. The glow discharge inception and post-discharge relaxation of charged and neutral active particles in synthetic air at low pressure

    NASA Astrophysics Data System (ADS)

    Jovanović, A. P.; Marković, V. Lj; Stamenković, S. N.; Stankov, M. N.

    2015-11-01

    The study of dc glow discharge inception and post-discharge relaxation of charged and neutral active particles in synthetic air at low pressure is presented. The breakdown time delay dependence as a function of relaxation time \\overline{{{t}\\text{d}}}(τ ) (the memory curve) is measured and modelled from milliseconds to the saturation region determined by the cosmic rays and natural radioactivity level. Due to fast conversion \\text{N}2++{{\\text{O}}2}\\to {{\\text{N}}2}+\\text{O}2+ , relaxation of dc discharge in synthetic air in the time interval from one to about ninety milliseconds is dominated by the diffusion decay of molecular oxygen {{O}}_2^ + ions. The change of regimes, from ambipolar to the free diffusion limit, is investigated and the variation of effective diffusion coefficients is determined. The late relaxation is explained by the kinetics of nitrogen atoms, recombining on the surfaces of gas discharge tube and stainless steel electrodes and relevant surface recombination coefficients are determined.

  5. Generation and reactivity of putative support systems, Ce-Al neutral binary oxide nanoclusters: CO oxidation and C–H bond activation

    SciTech Connect

    Wang, Zhe-Chen; Yin, Shi; Bernstein, Elliot R.

    2013-11-21

    Both ceria (CeO{sub 2}) and alumina (Al{sub 2}O{sub 3}) are very important catalyst support materials. Neutral binary oxide nanoclusters (NBONCs), Ce{sub x}Al{sub y}O{sub z}, are generated and detected in the gas phase and their reactivity with carbon monoxide (CO) and butane (C{sub 4}H{sub 10}) is studied. The very active species CeAlO{sub 4}{sup •} can react with CO and butane via O atom transfer (OAT) and H atom transfer (HAT), respectively. Other Ce{sub x}Al{sub y}O{sub z} NBONCs do not show reactivities toward CO and C{sub 4}H{sub 10}. The structures, as well as the reactivities, of Ce{sub x}Al{sub y}O{sub z} NBONCs are studied theoretically employing density functional theory (DFT) calculations. The ground state CeAlO{sub 4}{sup •} NBONC possesses a kite-shaped structure with an O{sub t}CeO{sub b}O{sub b}AlO{sub t} configuration (O{sub t}, terminal oxygen; O{sub b}, bridging oxygen). An unpaired electron is localized on the O{sub t} atom of the AlO{sub t} moiety rather than the CeO{sub t} moiety: this O{sub t} centered radical moiety plays a very important role for the reactivity of the CeAlO{sub 4}{sup •} NBONC. The reactivities of Ce{sub 2}O{sub 4}, CeAlO{sub 4}{sup •}, and Al{sub 2}O{sub 4} toward CO are compared, emphasizing the importance of a spin-localized terminal oxygen for these reactions. Intramolecular charge distributions do not appear to play a role in the reactivities of these neutral clusters, but could be important for charged isoelectronic BONCs. DFT studies show that the reaction of CeAlO{sub 4}{sup •} with C{sub 4}H{sub 10} to form the CeAlO{sub 4}H•C{sub 4}H{sub 9}{sup •} encounter complex is barrierless. While HAT processes have been previously characterized for cationic and anionic oxide clusters, the reported study is the first observation of a HAT process supported by a ground state neutral oxide cluster. Mechanisms for catalytic oxidation of CO over surfaces of Al{sub x}O{sub y}/M{sub m}O{sub n} or M{sub m}O{sub n

  6. Monohydroxylated metabolites of the K2 synthetic cannabinoid JWH-073 retain intermediate to high cannabinoid 1 receptor (CB1R) affinity and exhibit neutral antagonist to partial agonist activity.

    PubMed

    Brents, Lisa K; Gallus-Zawada, Anna; Radominska-Pandya, Anna; Vasiljevik, Tamara; Prisinzano, Thomas E; Fantegrossi, William E; Moran, Jeffery H; Prather, Paul L

    2012-04-01

    K2 and several similar purported "incense products" spiked with synthetic cannabinoids are abused as cannabis substitutes. We hypothesized that metabolism of JWH-073, a prevalent cannabinoid found in K2, contributes to toxicity associated with K2 use. Competition receptor binding studies and G-protein activation assays, both performed by employing mouse brain homogenates, were used to determine the affinity and intrinsic activity, respectively, of potential monohydroxylated (M1, M3-M5) and monocarboxylated (M6) metabolites at cannabinoid 1 receptors (CB1Rs). Surprisingly, M1, M4 and M5 retain nanomolar affinity for CB1Rs, while M3 displays micromolar affinity and M6 does not bind to CB1Rs. JWH-073 displays equivalent efficacy to that of the CB1R full agonist CP-55,940, while M1, M3, and M5 act as CB1R partial agonists, and M4 shows little or no intrinsic activity. Further in vitro investigation by Schild analysis revealed that M4 acts as a competitive neutral CB1R antagonist (K(b)∼40nM). In agreement with in vitro studies, M4 also demonstrates CB1R antagonism in vivo by blunting cannabinoid-induced hypothermia in mice. Interestingly, M4 does not block agonist-mediated responses of other measures in the cannabinoid tetrad (e.g., locomotor suppression, catalepsy or analgesia). Finally, also as predicted by in vitro results, M1 exhibits agonist activity in vivo by inducing significant hypothermia and suppression of locomotor activity in mice. In conclusion, the present study indicates that further work examining the physiological effects of synthetic cannabinoid metabolism is warranted. Such a complex mix of metabolically produced CB1R ligands may contribute to the adverse effect profile of JWH-073-containing products.

  7. Monohydroxylated metabolites of the K2 synthetic cannabinoid JWH-073 retain intermediate to high cannabinoid 1 receptor (CB1R) affinity and exhibit neutral antagonist to partial agonist activity

    PubMed Central

    Brents, Lisa K.; Gallus-Zawada, Anna; Radominska-Pandya, Anna; Vasiljevik, Tamara; Prisinzano, Thomas E.; Fantegrossi, William E.; Moran, Jeffery H.; Prather, Paul L.

    2012-01-01

    K2 and several similar purported “incense products” spiked with synthetic cannabinoids are abused as cannabis substitutes. We hypothesized that metabolism of JWH-073, a prevalent cannabinoid found in K2, contributes to toxicity associated with K2 use. Competition receptor binding studies and G-protein activation assays, both performed by employing mouse brain homogenates, were used to determine the affinity and intrinsic activity, respectively, of potential monohydroxylated (M1, M3–M5) and monocarboxylated (M6) metabolites at cannabinoid 1 receptors (CB1Rs). Surprisingly, M1, M4 and M5 retain nanomolar affinity for CB1Rs, while M3 displays micromolar affinity and M6 does not bind to CB1Rs. JWH-073 displays equivalent efficacy to that of the CB1R full agonist CP-55,940, while M1, M3, and M5 act as CB1R partial agonists, and M4 shows little or no intrinsic activity. Further in vitro investigation by Schild analysis revealed that M4 acts as a competitive neutral CB1R antagonist (Kb~40nM). In agreement with in vitro studies, M4 also demonstrates CB1R antagonism in vivo by blunting cannabinoid-induced hypothermia in mice. Interestingly, M4 does not block agonist-mediated responses of other measures in the cannabinoid tetrad (e.g., locomotor suppression, catalepsy or analgesia). Finally, also as predicted by in vitro results, M1 exhibits agonist activity in vivo by inducing significant hypothermia and suppression of locomotor activity in mice. In conclusion, the present study indicates that further work examining the physiological effects of synthetic cannabinoid metabolism is warranted. Such a complex mix of metabolically produced CB1R ligands may contribute to the adverse effect profile of JWH-073-containing products. PMID:22266354

  8. Neutron Activation Analysis of Water - A Review

    NASA Technical Reports Server (NTRS)

    Buchanan, John D.

    1971-01-01

    Recent developments in this field are emphasized. After a brief review of basic principles, topics discussed include sources of neutrons, pre-irradiation physical and chemical treatment of samples, neutron capture and gamma-ray analysis, and selected applications. Applications of neutron activation analysis of water have increased rapidly within the last few years and may be expected to increase in the future.

  9. International Thermonuclear Experimental Reactor (ITER) neutral beam design

    SciTech Connect

    Myers, T.J.; Brook, J.W.; Spampinato, P.T.; Mueller, J.P.; Luzzi, T.E.; Sedgley, D.W. . Space Systems Div.)

    1990-10-01

    This report discusses the following topics on ITER neutral beam design: ion dump; neutralizer and module gas flow analysis; vacuum system; cryogenic system; maintainability; power distribution; and system cost.

  10. Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle: Mechanistic insights into outer-membrane permeabilization and endotoxin neutralization.

    PubMed

    Saravanan, Rathi; Mohanram, Harini; Joshi, Mangesh; Domadia, Prerna N; Torres, Jaume; Ruedl, Christiane; Bhattacharjya, Surajit

    2012-07-01

    Tachyplesin-1, a disulfide stabilized beta-hairpin antimicrobial peptide, can be found at the hemocytes of horse shoe crab Tachypleus tridentatus. A cysteine deleted linear analog of tachyplesin-1 or CDT (KWFRVYRGIYRRR-NH2) contains a broad spectrum of bactericidal activity with a reduced hemolytic property. The bactericidal activity of CDT stems from selective interactions with the negatively charged lipids including LPS. In this work, CDT-LPS interactions were investigated using NMR spectroscopy, optical spectroscopy and functional assays. We found that CDT neutralized LPS and disrupted permeability barrier of the outer membrane. Zeta potential and ITC studies demonstrated charge compensation and hydrophobic interactions of CDT with the LPS-outer membrane, respectively. Secondary structure of the peptide was probed by CD and FT-IR experiments indicating beta-strands and/or beta-turn conformations in the LPS micelle. An ensemble of structures, determined in LPS micelle by NMR, revealed a beta-hairpin like topology of the CDT peptide that was typified by an extended cationic surface and a relatively shorter segment of hydrophobic region. Interestingly, at the non-polar face, residue R11 was found to be in a close proximity to the indole ring of W2, suggesting a cation-n type interactions. Further, saturation transfer difference (STD) NMR studies established intimate contacts among the aromatic and cationic residues of CDT with the LPS micelle. Fluorescence and dynamic light scattering experiments demonstrated that CDT imparted structural destabilization to the aggregated states of LPS. Collectively, atomic resolution structure and interactions of CDT with the outer membrane-LPS could be exploited for developing potent broad spectrum antimicrobial and anti-sepsis agents. PMID:22464970

  11. Dual inhibition of angiotensin-converting enzyme and neutral endopeptidase by the orally active inhibitor mixanpril: a potential therapeutic approach in hypertension.

    PubMed Central

    Fournié-Zaluski, M C; Gonzalez, W; Turcaud, S; Pham, I; Roques, B P; Michel, J B

    1994-01-01

    In the treatment of cardiovascular disease, it could be of therapeutic interest to associate the hypotensive effects due to the inhibition of angiotensin II formation with the diuretic and natriuretic responses induced by the protection of the endogenous atrial natriuretic peptide (ANP). Investigation of this hypothesis requires an orally active compound able to simultaneously inhibit angiotensin-converting enzyme (ACE) and neutral endopeptidase (NEP), which is involved in renal ANP metabolism. Such compounds have been rationally designed by taking into account the structural characteristics of the active site of both peptidases. Among them, RB 105, N-[(2S,3R)-2-mercaptomethyl-1-oxo-3-phenylbutyl]-(S)-alanine, inhibited NEP and ACE with Ki values of 1.7 +/- 0.3 nM and 4.2 +/- 0.5 nM, respectively. Intravenous infusion of RB 105 in conscious spontaneously hypertensive rats prevented the pressor response to exogenous angiotensin I and potentiated the natriuretic response to ANP. Infusion of RB 105, at 2.5, 5, 10, 25, and 50 mg/kg per hr decreased blood pressure dose-dependently in conscious catheterized spontaneously hypertensive rats and increased diuresis and natriuresis. Infusion of RB 105 as a bolus of 25 mg/kg followed by 25 mg/kg per hr similarly decreased blood pressure and increased natriuresis in three different models of hypertension (renovascular, deoxycorticosterone acetate-salt, and spontaneously hypertensive rats). Mixanpril, a lipophilic prodrug of RB 105 (ED50 values when given orally to mice, 0.7 mg/kg for NEP; 7 mg/kg for ACE), elicited dose-dependent hypotensive effects of long duration in spontaneously hypertensive rats after oral administration [-37 mmHg for 50 mg/kg twice a day (1 mmHg = 133 Pa) and is therefore the first dual NEP/ACE inhibitor potentially useful for clinical investigations. Images PMID:8171037

  12. Humanized-single domain antibodies (VH/VHH) that bound specifically to Naja kaouthia phospholipase A2 and neutralized the enzymatic activity.

    PubMed

    Chavanayarn, Charnwit; Thanongsaksrikul, Jeeraphong; Thueng-In, Kanyarat; Bangphoomi, Kunan; Sookrung, Nitat; Chaicumpa, Wanpen

    2012-07-01

    Naja kaouthia (monocled cobra) venom contains many isoforms of secreted phospholipase A2 (sPLA(2)). The PLA(2) exerts several pharmacologic and toxic effects in the snake bitten subject, dependent or independent on the enzymatic activity. N. kaouthia venom appeared in two protein profiles, P3 and P5, after fractionating the venom by ion exchange column chromatography. In this study, phage clones displaying humanized-camel single domain antibodies (VH/V(H)H) that bound specifically to the P3 and P5 were selected from a humanized-camel VH/V(H)H phage display library. Two phagemid transfected E. coli clones (P3-1 and P3-3) produced humanized-V(H)H, while another clone (P3-7) produced humanized-VH. At the optimal venom:antibody ratio, the VH/V(H)H purified from the E. coli homogenates neutralized PLA(2) enzyme activity comparable to the horse immune serum against the N. kaouthia holo-venom. Homology modeling and molecular docking revealed that the VH/V(H)H covered the areas around the PLA(2) catalytic groove and inserted their Complementarity Determining Regions (CDRs) into the enzymatic cleft. It is envisaged that the VH/V(H)H would ameliorate/abrogate the principal toxicity of the venom PLA(2) (membrane phospholipid catabolism leading to cellular and subcellular membrane damage which consequently causes hemolysis, hemorrhage, and dermo-/myo-necrosis), if they were used for passive immunotherapy of the cobra bitten victim. The speculation needs further investigations.

  13. Humanized-single domain antibodies (VH/VHH) that bound specifically to Naja kaouthia phospholipase A2 and neutralized the enzymatic activity.

    PubMed

    Chavanayarn, Charnwit; Thanongsaksrikul, Jeeraphong; Thueng-In, Kanyarat; Bangphoomi, Kunan; Sookrung, Nitat; Chaicumpa, Wanpen

    2012-07-01

    Naja kaouthia (monocled cobra) venom contains many isoforms of secreted phospholipase A2 (sPLA(2)). The PLA(2) exerts several pharmacologic and toxic effects in the snake bitten subject, dependent or independent on the enzymatic activity. N. kaouthia venom appeared in two protein profiles, P3 and P5, after fractionating the venom by ion exchange column chromatography. In this study, phage clones displaying humanized-camel single domain antibodies (VH/V(H)H) that bound specifically to the P3 and P5 were selected from a humanized-camel VH/V(H)H phage display library. Two phagemid transfected E. coli clones (P3-1 and P3-3) produced humanized-V(H)H, while another clone (P3-7) produced humanized-VH. At the optimal venom:antibody ratio, the VH/V(H)H purified from the E. coli homogenates neutralized PLA(2) enzyme activity comparable to the horse immune serum against the N. kaouthia holo-venom. Homology modeling and molecular docking revealed that the VH/V(H)H covered the areas around the PLA(2) catalytic groove and inserted their Complementarity Determining Regions (CDRs) into the enzymatic cleft. It is envisaged that the VH/V(H)H would ameliorate/abrogate the principal toxicity of the venom PLA(2) (membrane phospholipid catabolism leading to cellular and subcellular membrane damage which consequently causes hemolysis, hemorrhage, and dermo-/myo-necrosis), if they were used for passive immunotherapy of the cobra bitten victim. The speculation needs further investigations. PMID:22852068

  14. Measurement of the neutral D meson mixing parameters in a time-dependent amplitude analysis of the D0→π+π-π0 decay

    NASA Astrophysics Data System (ADS)

    Lees, J. P.; Poireau, V.; Tisserand, V.; Grauges, E.; Palano, A.; Eigen, G.; Brown, D. N.; Kolomensky, Yu. G.; Koch, H.; Schroeder, T.; Hearty, C.; Mattison, T. S.; McKenna, J. A.; So, R. Y.; Blinov, V. E.; Buzykaev, A. R.; Druzhinin, V. P.; Golubev, V. B.; Kravchenko, E. A.; Onuchin, A. P.; Serednyakov, S. I.; Skovpen, Yu. I.; Solodov, E. P.; Todyshev, K. Yu.; Lankford, A. J.; Gary, J. W.; Long, O.; Eisner, A. M.; Lockman, W. S.; Panduro Vazquez, W.; Chao, D. S.; Cheng, C. H.; Echenard, B.; Flood, K. T.; Hitlin, D. G.; Kim, J.; Miyashita, T. S.; Ongmongkolkul, P.; Porter, F. C.; Röhrken, M.; Huard, Z.; Meadows, B. T.; Pushpawela, B. G.; Sokoloff, M. D.; Sun, L.; Smith, J. G.; Wagner, S. R.; Bernard, D.; Verderi, M.; Bettoni, D.; Bozzi, C.; Calabrese, R.; Cibinetto, G.; Fioravanti, E.; Garzia, I.; Luppi, E.; Santoro, V.; Calcaterra, A.; de Sangro, R.; Finocchiaro, G.; Martellotti, S.; Patteri, P.; Peruzzi, I. M.; Piccolo, M.; Zallo, A.; Passaggio, S.; Patrignani, C.; Bhuyan, B.; Mallik, U.; Chen, C.; Cochran, J.; Prell, S.; Ahmed, H.; Gritsan, A. V.; Arnaud, N.; Davier, M.; Le Diberder, F.; Lutz, A. M.; Wormser, G.; Lange, D. J.; Wright, D. M.; Coleman, J. P.; Gabathuler, E.; Hutchcroft, D. E.; Payne, D. J.; Touramanis, C.; Bevan, A. J.; Di Lodovico, F.; Sacco, R.; Cowan, G.; Banerjee, Sw.; Brown, D. N.; Davis, C. L.; Denig, A. G.; Fritsch, M.; Gradl, W.; Griessinger, K.; Hafner, A.; Schubert, K. R.; Barlow, R. J.; Lafferty, G. D.; Cenci, R.; Jawahery, A.; Roberts, D. A.; Cowan, R.; Cheaib, R.; Robertson, S. H.; Dey, B.; Neri, N.; Palombo, F.; Cremaldi, L.; Godang, R.; Summers, D. J.; Taras, P.; De Nardo, G.; Sciacca, C.; Raven, G.; Jessop, C. P.; LoSecco, J. M.; Honscheid, K.; Kass, R.; Gaz, A.; Margoni, M.; Posocco, M.; Rotondo, M.; Simi, G.; Simonetto, F.; Stroili, R.; Akar, S.; Ben-Haim, E.; Bomben, M.; Bonneaud, G. R.; Calderini, G.; Chauveau, J.; Marchiori, G.; Ocariz, J.; Biasini, M.; Manoni, E.; Rossi, A.; Batignani, G.; Bettarini, S.; Carpinelli, M.; Casarosa, G.; Chrzaszcz, M.; Forti, F.; Giorgi, M. A.; Lusiani, A.; Oberhof, B.; Paoloni, E.; Rama, M.; Rizzo, G.; Walsh, J. J.; Smith, A. J. S.; Anulli, F.; Faccini, R.; Ferrarotto, F.; Ferroni, F.; Pilloni, A.; Piredda, G.; Bünger, C.; Dittrich, S.; Grünberg, O.; Heß, M.; Leddig, T.; Voß, C.; Waldi, R.; Adye, T.; Wilson, F. F.; Emery, S.; Vasseur, G.; Aston, D.; Cartaro, C.; Convery, M. R.; Dorfan, J.; Dunwoodie, W.; Ebert, M.; Field, R. C.; Fulsom, B. G.; Graham, M. T.; Hast, C.; Innes, W. R.; Kim, P.; Leith, D. W. G. S.; Luitz, S.; Luth, V.; MacFarlane, D. B.; Muller, D. R.; Neal, H.; Ratcliff, B. N.; Roodman, A.; Sullivan, M. K.; Va'vra, J.; Wisniewski, W. J.; Purohit, M. V.; Wilson, J. R.; Randle-Conde, A.; Sekula, S. J.; Bellis, M.; Burchat, P. R.; Puccio, E. M. T.; Alam, M. S.; Ernst, J. A.; Gorodeisky, R.; Guttman, N.; Peimer, D. R.; Soffer, A.; Spanier, S. M.; Ritchie, J. L.; Schwitters, R. F.; Izen, J. M.; Lou, X. C.; Bianchi, F.; De Mori, F.; Filippi, A.; Gamba, D.; Lanceri, L.; Vitale, L.; Martinez-Vidal, F.; Oyanguren, A.; Albert, J.; Beaulieu, A.; Bernlochner, F. U.; King, G. J.; Kowalewski, R.; Lueck, T.; Nugent, I. M.; Roney, J. M.; Tasneem, N.; Gershon, T. J.; Harrison, P. F.; Latham, T. E.; Prepost, R.; Wu, S. L.; BaBar Collaboration

    2016-06-01

    We perform the first measurement on the D0-D¯ 0 mixing parameters using a time-dependent amplitude analysis of the decay D0→π+π-π0 . The data were recorded with the BABAR detector at center-of-mass energies at and near the ϒ (4 S ) resonance, and correspond to an integrated luminosity of approximately 468.1 fb-1 . The neutral D meson candidates are selected from D*(2010 )+→D0πs+ decays where the flavor at the production is identified by the charge of the low-momentum pion, πs+ . The measured mixing parameters are x =(1.5 ±1.2 ±0.6 )% and y =(0.2 ±0.9 ±0.5 )% , where the quoted uncertainties are statistical and systematic, respectively.

  15. Ion densities and composition of Titan's upper atmosphere derived from the Cassini Ion Neutral Mass Spectrometer: Analysis methods and comparison of measured ion densities to photochemical model simulations

    NASA Astrophysics Data System (ADS)

    Mandt, Kathleen E.; Gell, David A.; Perry, Mark; Hunter Waite, J., Jr.; Crary, Frank A.; Young, David; Magee, Brian A.; Westlake, Joseph H.; Cravens, Thomas; Kasprzak, Wayne; Miller, Greg; Wahlund, Jan-Erik; Ågren, Karin; Edberg, Niklas J. T.; Heays, Alan N.; Lewis, Brenton R.; Gibson, Stephen T.; de la Haye, V.; Liang, Mao-Chang

    2012-10-01

    The Cassini Ion Neutral Mass Spectrometer (INMS) has measured both neutral and ion species in Titan's upper atmosphere and ionosphere and the Enceladus plumes. Ion densities derived from INMS measurements are essential data for constraining photochemical models of Titan's ionosphere. The objective of this paper is to present an optimized method for converting raw data measured by INMS to ion densities. To do this, we conduct a detailed analysis of ground and in-flight calibration to constrain the instrument response to ion energy, the critical parameter on which the calibration is based. Data taken by the Cassini Radio Plasma Wave Science Langmuir Probe and the Cassini Plasma Spectrometer Ion Beam Spectrometer are used as independent measurement constraints in this analysis. Total ion densities derived with this method show good agreement with these data sets in the altitude region (˜1100-1400 km) where ion drift velocities are low and the mass of the ions is within the measurement range of the INMS (1-99 Daltons). Although ion densities calculated by the method presented here differ slightly from those presented in previous INMS publications, we find that the implications for the science presented in previous publications is mostly negligible. We demonstrate the role of the INMS ion densities in constraining photochemical models and find that (1) cross sections having high resolution as a function of wavelength are necessary for calculating the initial photoionization products and (2) there are disagreements between the measured ion densities representative of the initial steps in Titan photochemistry that require further investigation.

  16. Two-Point Correlation Analysis Of Neutrally Stratified Flow Within And Above A Forest From Large-Eddy Simulation

    NASA Astrophysics Data System (ADS)

    Su, Hong-Bing; Shaw, Roger H.; Tha Paw U, Kyaw

    Two-point space-time correlations ofvelocities, a passive scalar and static pressure arecalculated using the resolvable flow fields computedby large-eddy simulation (LES) of neutrally stratifiedflow within and above a sparse forest. Zero-time-lagspatial auto-correlation contours in thestreamwise-vertical cross-section for longitudinal andlateral velocities and for a scalar are tilted fromthe vertical in the downstream direction, as istypical in near-wall sheared flow. On the other hand,auto-correlations of vertical velocity and of staticpressure are vertically coherent. Zero-time-lagspatial auto-correlations in the spanwise-verticalcross-section show no distinct tilt, and those forboth longitudinal and vertical velocities demonstratedistinct negative side lobes in the middle forest andabove, while longitudinal velocity in the subcrowntrunk space is laterally in-phase. Static pressureperturbations appear to be spatially coherent in thespanwise direction at all heights, especially insidethe forest. Near the forest floor, longitudinalvelocity is found to be in-phase with static pressureperturbation and to be closely linked to theinstantaneous streamwise pressure gradient, supportinga previous proposal that longitudinal velocity in thisregion is dominantly modulated by the pressurepatterns associated with the coherent sweep/ejectionevents. Near treetop height, a lack of linkage betweenthe pressure gradient and the local time derivative ofthe longitudinal velocity supports the hypothesis ofadvection dominating turbulent flow.The major phase characteristics of the two-pointcorrelations essentially remained the same from fourLES runs with different domain size and/or gridresolution. A larger LES domain yielded betteragreement with field observations in a real forest onboth the magnitudes of the correlations and thesingle-point integral time scales. A finer gridresolution in the LES led to a faster rate of decreaseof correlation with increasing separation in space ortime

  17. Neutrality in Language Policy

    ERIC Educational Resources Information Center

    Wee, Lionel

    2010-01-01

    The unavoidability of language makes it critical that language policies appeal to some notion of language neutrality as part of their rationale, in order to assuage concerns that the policies might otherwise be unduly discriminatory. However, the idea of language neutrality is deeply ideological in nature, since it is not only an attempt to treat…

  18. Self-neutralized ion beam

    SciTech Connect

    Salvadori, M. C.; Teixeira, F. S.; Nikolaev, A.; Savkin, K. P.; Oks, E. M.; Spaedtke, P.; Yu, K. M.; Brown, I. G.

    2011-10-15

    A vacuum arc ion source provides high current beams of metal ions that have been used both for accelerator injection and for ion implantation, and in both of these applications the degree of space charge neutralization of the beam is important. In accelerator injection application, the beam from the ion source may be accelerated further (post-acceleration), redirected by a bending magnet(s), or focused with magnetic or electrostatic lenses, and knowledge of the beam space charge is needed for optimal design of the optical elements. In ion implantation application, any build-up of positive charge in the insulating targets must be compensated by a simultaneous flux of cold electrons so as to provide overall charge neutrality of the target. We show that in line-of-sight ion implantation using a vacuum arc ion source, the high current ion beam carries along its own background sea of cold electrons, and this copious source of electrons provides a ''self-neutralizing'' feature to the beam. Here we describe experiments carried out in order to demonstrate this effect, and we provide an analysis showing that the beam is space-charge-neutralized to a very high degree.

  19. Human hair neutron activation analysis: Analysis on population level, mapping

    NASA Astrophysics Data System (ADS)

    Zhuk, L. I.; Kist, A. A.

    1999-01-01

    Neutron activation analysis is an outstanding analytical method having very wide applications in various fields. Analysis of human hair within last decades mostly based on neutron activation analysis is a very attractive illustration of the application of nuclear analytical techniques. Very interesting question is how the elemental composition differs in different areas or cities. In this connection the present paper gives average data and maps of various localities in the vicinity of drying-out Aral Sea and of various industrial cities in Central Asia.

  20. Degradation of 32 emergent contaminants by UV and neutral photo-fenton in domestic wastewater effluent previously treated by activated sludge.

    PubMed

    De la Cruz, N; Giménez, J; Esplugas, S; Grandjean, D; de Alencastro, L F; Pulgarín, C

    2012-04-15

    This study focuses on the removal of 32 selected micropollutants (pharmaceuticals, corrosion inhibitors and biocides/pesticides) found in an effluent coming from a municipal wastewater treatment plant (MWTP) based on activated sludge. Dissolved organic matter was present, with an initial total organic carbon of 15.9 mg L(-1), and a real global quantity of micropollutants of 29.5 μg L(-1). The treatments tested on the micropollutants removal were: UV-light emitting at 254 nm (UV(254)) alone, dark Fenton (Fe(2+,3+)/H(2)O(2)) and photo-Fenton (Fe(2+,3+)/H(2)O(2)/light). Different irradiation sources were used for the photo-Fenton experiences: UV(254) and simulated sunlight. Iron and H(2)O(2) concentrations were also changed in photo-Fenton experiences in order to evaluate its influence on the degradation. All the experiments were developed at natural pH, near neutral. Photo-Fenton treatments employing UV(254), 50 mg L(-1) of H(2)O(2), with and without adding iron (5 mg L(-1) of Fe(2+) added or 1.48 mg L(-1) of total iron already present) gave the best results. Global percentages of micropollutants removal achieved were 98 and a 97% respectively, after 30 min of treatments. As the H(2)O(2) concentration increased (10, 25 and 50 mg L(-1)), best degradations were observed. UV(254), Fenton, and photo-Fenton under simulated sunlight gave less promising results with lower percentages of removal. The highlight of this paper is to point out the possibility of the micropollutants degradation in spite the presence of DOM in much higher concentrations. PMID:22305640

  1. IgG subclass antibodies to human cytomegalovirus (CMV) in normal human plasma samples and immune globulins and their neutralizing activities.

    PubMed

    Gupta, C K; Leszczynski, J; Gupta, R K; Siber, G R

    1996-06-01

    An enzyme-linked immunoabsorbent assay (ELISA) was developed for quantitation of IgG subclass antibodies to human cytomegalovirus (CMV) in human serum or plasma samples and in immune globulin (IG) preparations. The assay was based on the parallel titration of known concentrations of purified IgG subclass myeloma proteins and a specific CMV antiserum. The purified IgG subclass myeloma proteins were captured on an ELISA plate pre-coated with anti-human kappa, anti-human lambda or a mixture of anti-human kappa and lambda antibodies and the specific antiserum was titrated against CMV antigen coated on the plate. IgG subclass antibodies, captured or bound to antigen, were quantitated with IGG subclass heavy chain specific monoclonal antibodies. The method was highly reproducible, specific and sensitive. Using this method, 257 human plasma samples and 50 IG preparations were assayed for CMV specific IgG subclass and IgM antibodies. The major IgG subclass antibody to CMV was IgG1 which represented more than 96% of CMV IgG antibodies, followed by IgG3 (mean CMV IgG3 antibody content was 3% of IgG antibodies in IG preparations and 1.8% in plasma samples). A majority of the samples had low levels of IgG2 antibodies and a few samples exhibited low levels of IgG4 antibodies. IG preparations showed very low levels of CMV IgM antibodies whereas plasma samples had 14.2% of CMV antibodies (IgG and IgM) as IgM antibodies. Virus neutralizing (Nt) activity of these samples showed a significant correlation with CMV IgG1 antibodies. Nine samples of plasma and IGs were further evaluated for Nt activity of IgG1 and IgG3 antibodies by separating IgG3 from the rest of the antibodies with protein A agarose. IgG3 antibodies showed much higher Nt activity than IgG1 antibodies suggesting that enrichment of IgG3 antibodies in IG preparations may be useful in preparing CMV specific IG.

  2. A new EV71 VP3 epitope in norovirus P particle vector displays neutralizing activity and protection in vivo in mice.

    PubMed

    Jiang, Liping; Fan, Rongjun; Sun, Shiyang; Fan, Peihu; Su, Weiheng; Zhou, Yan; Gao, Feng; Xu, Fei; Kong, Wei; Jiang, Chunlai

    2015-11-27

    Enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16), as the main agents causing hand, foot and mouth disease (HFMD), have become a serious public health concern in the Asia-Pacific region. Recently, various neutralizing B cell epitopes of EV71 were identified as targets for promising vaccine candidates. Structural studies of Picornaviridae indicated that potent immunodominant epitopes typically lie in the hypervariable loop of capsid surfaces. However, cross-neutralizing antibodies and cross-protection between EV71 and CVA16 have not been observed. Therefore, we speculated that divergent sequences of the two viruses are key epitopes for inducing protective neutralizing responses. In this study, we selected 10 divergent epitope candidates based on alignment of the EV71 and CVA16 P1 amino acid sequences using the Multalin interface page, and these epitopes are conserved among all subgenotypes of EV71. Simultaneously, by utilizing the norovirus P particle as a novel vaccine delivery carrier, we identified the 71-6 epitope (amino acid 176-190 of VP3) as a conformational neutralizing epitope against EV71 in an in vitro micro-neutralization assay as well as an in vivo protection assay in mice. Altogether, these results indicated that the incorporation of the 71-6 epitope into the norovirus P domain can provide a promising candidate for an effective synthetic peptide-based vaccine against EV71.

  3. A new EV71 VP3 epitope in norovirus P particle vector displays neutralizing activity and protection in vivo in mice.

    PubMed

    Jiang, Liping; Fan, Rongjun; Sun, Shiyang; Fan, Peihu; Su, Weiheng; Zhou, Yan; Gao, Feng; Xu, Fei; Kong, Wei; Jiang, Chunlai

    2015-11-27

    Enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16), as the main agents causing hand, foot and mouth disease (HFMD), have become a serious public health concern in the Asia-Pacific region. Recently, various neutralizing B cell epitopes of EV71 were identified as targets for promising vaccine candidates. Structural studies of Picornaviridae indicated that potent immunodominant epitopes typically lie in the hypervariable loop of capsid surfaces. However, cross-neutralizing antibodies and cross-protection between EV71 and CVA16 have not been observed. Therefore, we speculated that divergent sequences of the two viruses are key epitopes for inducing protective neutralizing responses. In this study, we selected 10 divergent epitope candidates based on alignment of the EV71 and CVA16 P1 amino acid sequences using the Multalin interface page, and these epitopes are conserved among all subgenotypes of EV71. Simultaneously, by utilizing the norovirus P particle as a novel vaccine delivery carrier, we identified the 71-6 epitope (amino acid 176-190 of VP3) as a conformational neutralizing epitope against EV71 in an in vitro micro-neutralization assay as well as an in vivo protection assay in mice. Altogether, these results indicated that the incorporation of the 71-6 epitope into the norovirus P domain can provide a promising candidate for an effective synthetic peptide-based vaccine against EV71. PMID:26529072

  4. Analysis of cross-reactive antibodies recognizing the fusion loop of envelope protein and correlation with neutralizing antibody titers in Nicaraguan dengue cases.

    PubMed

    Lai, Chih-Yun; Williams, Katherine L; Wu, Yi-Chieh; Knight, Sarah; Balmaseda, Angel; Harris, Eva; Wang, Wei-Kung

    2013-01-01

    Dengue virus (DENV) is the leading cause of arboviral diseases in humans worldwide. The envelope (E) protein of DENV is the major target of neutralizing antibodies (Abs). Previous studies have shown that a significant proportion of anti-E Abs in human serum after DENV infection recognize the highly conserved fusion loop (FL) of E protein. The role of anti-FL Abs in protection against subsequent DENV infection versus pathogenesis remains unclear. A human anti-E monoclonal Ab was used as a standard in a virion-capture ELISA to measure the concentration of anti-E Abs, [anti-E Abs], in dengue-immune sera from Nicaraguan patients collected 3, 6, 12 and 18 months post-infection. The proportion of anti-FL Abs was determined by capture ELISA using virus-like particles containing mutations in FL, and the concentration of anti-FL Abs, [anti-FL Abs], was calculated. Neutralization titers (NT50) were determined using a previously described flow cytometry-based assay. Analysis of sequential samples from 10 dengue patients revealed [anti-E Abs] and [anti-FL Abs] were higher in secondary than in primary DENV infections. While [anti-FL Abs] did not correlate with NT50 against the current infecting serotype, it correlated with NT50 against the serotypes to which patients had likely not yet been exposed ("non-exposed" serotypes) in 14 secondary DENV3 and 15 secondary DENV2 cases. These findings demonstrate the kinetics of anti-FL Abs and provide evidence that anti-FL Abs play a protective role against "non-exposed" serotypes after secondary DENV infection. PMID:24069496

  5. Radioactivity analysis in niobium activation foils

    SciTech Connect

    Mueller, G.E.

    1995-06-01

    The motivation for this study was to measure and analyze the activity of six (6) niobium (Nb) foils (the x-rays from an internal transition in Nb-93m) and apply this information with previously obtained activation foil data. The niobium data was used to determine the epithermal to MeV range for the neutron spectrum and fluence. The foil activation data was re-evaluated in a spectrum analysis code (STAY`SL) to provide new estimates of the exposure at the Los Alamos Spallation Radiation Effect Facility (LASREF). The activity of the niobium foils was measured and analyzed at the University of Missouri-Columbia (UMC) under the direction of Professor William Miller. The spectrum analysis was performed at the University of Missouri-Rolla (UMR) by Professor Gary Mueller.

  6. Modelling the Neutral Sodium Tails of Comets

    NASA Astrophysics Data System (ADS)

    Birkett, K. S.; Jones, G. H.; Coates, A. J.

    2014-12-01

    Neutral sodium is typically easy to detect in active comets around perihelion, due to the very high efficiency of the sodium D transition, and at some comets a distinct neutral sodium tail is observed. The first distinct neutral sodium tail images were apparent in comet Hale-Bopp (C/1995 O1) data taken using CoCam [Cremonese et al, 1997], but since this initial detection similar features have been observed at a number of near-Sun comets using the SOHO/LASCO coronagraph. An understanding of the distribution and evolution of neutral cometary sodium may best be developed using a combination of spectra and images in different filters at multiple times throughout a comet's orbit. At present the source of neutral sodium in comets is unknown, primarily because the evolution of neutral cometary sodium is difficult to intuitively predict due to the Swings and Greenstein effects. Several authors [review presented in Cremonese et al, 1999] have suggested various combinations of sources of neutral sodium in the nuclear region, near-nuclear region, dust tail and ion tail. In order to understand the wide variety of cometary observations of neutral sodium available we have developed the first fully three dimensional, heliocentric distance dependent, versatile Monte Carlo neutral sodium tail model (initially based on a model developed by [Brown et al, 1998]). Our model is known as COMPASS (Cometary Orbital Motion at Perihelion: an Adaptable Sodium Simulation), and incorporates the unintuitive variation in radiation pressure influences on sodium atoms with different heliocentric velocities. We present the initial results of a comparison between COMPASS and observational data. We have found good agreement between the overall morphology of the neutral sodium tail imaged at comet Hale-Bopp and COMPASS, and have begun to extend the study to other comets of interest. We also present a comparison between simulated COMPASS spectra and observations. The versatility of COMPASS allows it to

  7. Neutral Buoyancy Simulator (NBS)

    NASA Technical Reports Server (NTRS)

    1972-01-01

    This is a cutaway illustration of the Neutral Buoyancy Simulator (NBS) at the Marshall Space Flight Center (MSFC ). The MSFC NBS provided an excellent environment for testing hardware to examine how it would operate in space and for evaluating techniques for space construction and spacecraft servicing. Here, engineers, designers, and astronauts performed various tests to develop basic concepts, preliminary designs, final designs, and crew procedures. The NBS was constructed of welded steel with polyester-resin coating. The water tank was 75-feet (22.9- meters) in diameter, 40-feet (12.2-meters) deep, and held 1.32 million gallons of water. Since it opened for operation in 1968, the NBS had supported a number of successful space missions, such as the Skylab, Solar Maximum Mission Satellite, Marned Maneuvering Unit, Experimental Assembly of Structures in Extravehicular Activity/Assembly Concept for Construction of Erectable Space Structures (EASE/ACCESS), the Hubble Space Telescope, and the Space Station. The function of the MSFC NBS was moved to the larger simulator at the Johnson Space Center and is no longer operational.

  8. Neutral Polymer Micelle Carriers with pH-Responsive, Endosome-Releasing Activity Modulate Antigen Trafficking to Enhance CD8 T-Cell Responses

    PubMed Central

    Keller, Salka; Wilson, John T; Patilea, Gabriela I; Kern, Hanna B; Convertine, Anthony J; Stayton, Patrick S

    2014-01-01

    Synthetic subunit vaccines need to induce CD8+ cytotoxic T-cell (CTL) responses for effective vaccination against intracellular pathogens. Most subunit vaccines primarily generate humoral immune responses, with a weaker than desired CD8+ cytotoxic T-cell response. Here, a neutral, pH-responsive polymer micelle carrier that alters intracellular antigen trafficking was shown to enhance CD8+ T-cell responses with a correlated increase in cytosolic delivery and a decrease in exocytosis. Polymer diblock carriers consisted of a N-(2-hydroxypropyl) methacrylamide corona block with pendant pyridyl disulfide groups for reversible conjugation of thiolated ovalbumin, and a tercopolymer ampholytic core-forming block composed of propylacrylic acid (PAA), dimethylaminoethyl methacrylate (DMAEMA), and butyl methacrylate (BMA). The diblock copolymers self-assembled into 25–30 nm diameter micellar nanoparticles. Conjugation of ovalbumin to the micelles significantly enhanced antigen cross-presentation in vitro relative to free ovalbumin, an unconjugated physical mixture of ovalbumin and polymer, and a non pH-responsive micelle-ovalbumin control. Mechanistic studies in a murine dendritic cell line (DC2.4) demonstrated micelle-mediated enhancements in intracellular antigen retention and cytosolic antigen accumulation. Approximately 90% of initially internalized ovalbumin-conjugated micelles were retained in cells after 1.5 h, compared to only ~40% for controls. Furthermore, cells dosed with conjugates displayed 67-fold higher cytosolic antigen levels relative to soluble ovalbumin 4 h post uptake. Subcutaneous immunization of mice with ovalbumin-polymer conjugates significantly enhanced antigen-specific CD8+ T cell responses (0.4 % IFN-γ+ of CD8+) compared to immunization with soluble protein, ovalbumin and polymer mixture, and the control micelle without endosome-releasing activity. Additionally, pH-responsive carrier facilitated antigen delivery to antigen presenting cells in the

  9. Neutral polymer micelle carriers with pH-responsive, endosome-releasing activity modulate antigen trafficking to enhance CD8(+) T cell responses.

    PubMed

    Keller, Salka; Wilson, John T; Patilea, Gabriela I; Kern, Hanna B; Convertine, Anthony J; Stayton, Patrick S

    2014-10-10

    Synthetic subunit vaccines need to induce CD8(+) cytotoxic T cell (CTL) responses for effective vaccination against intracellular pathogens. Most subunit vaccines primarily generate humoral immune responses, with a weaker than desired CD8(+) cytotoxic T cell response. Here, a neutral, pH-responsive polymer micelle carrier that alters intracellular antigen trafficking was shown to enhance CD8(+) T cell responses with a correlated increase in cytosolic delivery and a decrease in exocytosis. Polymer diblock carriers consisted of a N-(2-hydroxypropyl) methacrylamide corona block with pendent pyridyl disulfide groups for reversible conjugation of thiolated ovalbumin, and a tercopolymer ampholytic core-forming block composed of propylacrylic acid (PAA), dimethylaminoethyl methacrylate (DMAEMA), and butyl methacrylate (BMA). The diblock copolymers self-assembled into 25-30nm diameter micellar nanoparticles. Conjugation of ovalbumin to the micelles significantly enhanced antigen cross-presentation in vitro relative to free ovalbumin, an unconjugated physical mixture of ovalbumin and polymer, and a non-pH-responsive micelle-ovalbumin control. Mechanistic studies in a murine dendritic cell line (DC 2.4) demonstrated micelle-mediated enhancements in intracellular antigen retention and cytosolic antigen accumulation. Approximately 90% of initially internalized ovalbumin-conjugated micelles were retained in cells after 1.5h, compared to only ~40% for controls. Furthermore, cells dosed with conjugates displayed 67-fold higher cytosolic antigen levels relative to soluble ovalbumin 4h post uptake. Subcutaneous immunization of mice with ovalbumin-polymer conjugates significantly enhanced antigen-specific CD8(+) T cell responses (0.4% IFN-γ(+) of CD8(+)) compared to immunization with soluble protein, ovalbumin and polymer mixture, and the control micelle without endosome-releasing activity. Additionally, pH-responsive carrier facilitated antigen delivery to antigen presenting cells

  10. Solar Neutral Particles

    NASA Video Gallery

    This animation shows a neutral solar particle's path leaving the sun, following the magnetic field lines out to the heliosheath. The solar particle hits a hydrogen atom, stealing its electron, and ...

  11. Transforming Teacher Education, An Activity Theory Analysis

    ERIC Educational Resources Information Center

    McNicholl, Jane; Blake, Allan

    2013-01-01

    This paper explores the work of teacher education in England and Scotland. It seeks to locate this work within conflicting sociocultural views of professional practice and academic work. Drawing on an activity theory framework that integrates the analysis of these seemingly contradictory discourses with a study of teacher educators' practical…

  12. Computer-automated neutron activation analysis system

    SciTech Connect

    Minor, M.M.; Garcia, S.R.

    1983-01-01

    An automated delayed neutron counting and instrumental neutron activation analysis system has been developed at Los Alamos National Laboratory's Omega West Reactor (OWR) to analyze samples for uranium and 31 additional elements with a maximum throughput of 400 samples per day. 5 references.

  13. A METHOD FOR AUTOMATED ANALYSIS OF 10 ML WATER SAMPLES CONTAINING ACIDIC, BASIC, AND NEUTRAL SEMIVOLATILE COMPOUNDS LISTED IN USEPA METHOD 8270 BY SOLID PHASE EXTRACTION COUPLED IN-LINE TO LARGE VOLUME INJECTION GAS CHROMATOGRAPHY/MASS SPECTROMETRY

    EPA Science Inventory

    Data is presented showing the progress made towards the development of a new automated system combining solid phase extraction (SPE) with gas chromatography/mass spectrometry for the single run analysis of water samples containing a broad range of acid, base and neutral compounds...

  14. Spatial and temporal distribution of the neutral polymorphisms in the last ZFX intron: analysis of the haplotype structure and genealogy.

    PubMed Central

    Jaruzelska, J; Zietkiewicz, E; Batzer, M; Cole, D E; Moisan, J P; Scozzari, R; Tavaré, S; Labuda, D

    1999-01-01

    With 10 segregating sites (simple nucleotide polymorphisms) in the last intron (1089 bp) of the ZFX gene we have observed 11 haplotypes in 336 chromosomes representing a worldwide array of 15 human populations. Two haplotypes representing 77% of all chromosomes were distributed almost evenly among four continents. Five of the remaining haplotypes were detected in Africa and 4 others were restricted to Eurasia and the Americas. Using the information about the ancestral state of the segregating positions (inferred from human-great ape comparisons), we applied coalescent analysis to estimate the age of the polymorphisms and the resulting haplotypes. The oldest haplotype, with the ancestral alleles at all the sites, was observed at low frequency only in two groups of African origin. Its estimated age of 740 to 1100 kyr corresponded to the time to the most recent common ancestor. The two most frequent worldwide distributed haplotypes were estimated at 550 to 840 and 260 to 400 kyr, respectively, while the age of the continentally restricted polymorphisms was 120 to 180 kyr and smaller. Comparison of spatial and temporal distribution of the ZFX haplotypes suggests that modern humans diverged from the common ancestral stock in the Middle Paleolithic era. Subsequent range expansion prevented substantial gene flow among continents, separating African groups from populations that colonized Eurasia and the New World. PMID:10388827

  15. Evaluation of smallpox vaccines using variola neutralization.

    PubMed

    Damon, Inger K; Davidson, Whitni B; Hughes, Christine M; Olson, Victoria A; Smith, Scott K; Holman, Robert C; Frey, Sharon E; Newman, Frances; Belshe, Robert B; Yan, Lihan; Karem, Kevin

    2009-08-01

    The search for a 'third'-generation smallpox vaccine has resulted in the development and characterization of several vaccine candidates. A significant barrier to acceptance is the absence of challenge models showing induction of correlates of protective immunity against variola virus. In this light, virus neutralization provides one of few experimental methods to show specific 'in vitro' activity of vaccines against variola virus. Here, we provide characterization of the ability of a modified vaccinia virus Ankara vaccine to induce variola virus-neutralizing antibodies, and we provide comparison with the neutralization elicited by standard Dryvax vaccination.

  16. Neutron activation analysis of Etruscan pottery

    SciTech Connect

    Whitehead, J.; Silverman, A.; Ouellet, C.G.; Clark, D.D.; Hossain, T.Z

    1992-07-01

    Neutron activation analysis (NAA) has been widely used in archaeology for compositional analysis of pottery samples taken from sites of archaeological importance. Elemental profiles can determine the place of manufacture. At Cornell, samples from an Etruscan site near Siena, Italy, are being studied. The goal of this study is to compile a trace element concentration profile for a large number of samples. These profiles will be matched with an existing data bank in an attempt to understand the place of origin for these samples. The 500 kW TRIGA reactor at the Ward Laboratory is used to collect NAA data for these samples. Experiments were done to set a procedure for the neutron activation analysis with respect to sample preparation, selection of irradiation container, definition of activation and counting parameters and data reduction. Currently, we are able to analyze some 27 elements in samples of mass 500 mg with a single irradiation of 4 hours and two sequences of counting. Our sensitivity for many of the trace elements is better than 1 ppm by weight under the conditions chosen. In this talk, details of our procedure, including quality assurance as measured by NIST standard reference materials, will be discussed. In addition, preliminary results from data treatment using cluster analysis will be presented. (author)

  17. Structural Analysis of Human and Macaque mAbs 2909 and 2.5B: Implications for the Configuration of the Quaternary Neutralizing Epitope of HIV-1 gp120

    SciTech Connect

    Spurrier, Brett; Sampson, Jared M.; Totrov, Maxim; Li, Huiguang; O; Neal, Timothy; Williams, Constance; Robinson, James; Gorny, Miroslaw K.; Zolla-Pazner, Susan; Kong, Xiang-Peng

    2011-08-25

    The quaternary neutralizing epitope (QNE) of HIV-1 gp120 is preferentially expressed on the trimeric envelope spikes of intact HIV virions, and QNE-specific monoclonal antibodies (mAbs) potently neutralize HIV-1. Here, we present the crystal structures of the Fabs of human mAb 2909 and macaque mAb 2.5B. Both mAbs have long beta hairpin CDR H3 regions >20 {angstrom} in length that are each situated at the center of their respective antigen-binding sites. Computational analysis showed that the paratopes include the whole CDR H3, while additional CDR residues form shallow binding pockets. Structural modeling suggests a way to understand the configuration of QNEs and the antigen-antibody interaction for QNE mAbs. Our data will be useful in designing immunogens that may elicit potent neutralizing QNE Abs.

  18. Activity Analysis and Cost Analysis in Medical Schools.

    ERIC Educational Resources Information Center

    Koehler, John E.; Slighton, Robert L.

    There is no unique answer to the question of what an ongoing program costs in medical schools. The estimates of program costs generated by classical methods of cost accounting are unsatisfactory because such accounting cannot deal with the joint production or joint cost problem. Activity analysis models aim at calculating the impact of alternative…

  19. [Trace analysis of heavy metal ions in electroplate waste water by capillary electrophoresis with visual offline sample stacking via moving neutralization boundary].

    PubMed

    Fan, Yinping; Li, Shan; Fan, Liuyin; Cao, Chengxi

    2012-08-01

    A moving neutralization boundary (MNB) was developed as a novel model of visual offline sample stacking for the trace analysis of heavy metal ions (HMIs) by capillary zone electrophoresis (CZE). In the stacking system, the motion direction of MNB to cathode was used with 2.1 mmol/L HCl-98 mmol/L KCl-trace metal ions in the anodic solution and 4.0 mmol/L NaOH-96 mmol/L KCl in the cathodic solution. The voltage was constant at 180 V and the flow rate of the anolyte and catholyte was 1 mL/min. The metal ions in the gel after stacking were detected by capillary electrophoresis. The calibration curves showed good linear relationship (r > or = 0.998 5) in the concentration range used in the experiments. The pre-concentration factors were up to 80 - 150 and the limits of detection (LODs) were 0.163, 0.256, 0.077, 0.153, 0.203, 0.062 and 0.142 mg/L for Cu(II), Zn(II), Ni(II), Mg(II), Ca(II), Cr(III) and Fe(III), respectively, obviously lower than the national standards. The intra-day and inter-day assay precisions were good (the relative standard deviations (RSDs) less than 7.42%). Finally, the developed method has been successfully used for the stacking and the detection of heavy metal ions in electroplate waste water.

  20. A class of neutral functional differential equations.

    NASA Technical Reports Server (NTRS)

    Melvin, W. R.

    1972-01-01

    Formulation and study of the initial value problem for neutral functional differential equations. The existence, uniqueness, and continuation of solutions to this problem are investigated, and an analysis is made of the dependence of the solutions on the initial conditions and parameters, resulting in the derivation of a continuous dependence theorem in which the fundamental mathematical principles underlying the continuous dependence problem for a very general system of nonlinear neutral functional differential equations are separated out.

  1. Characterization of a Novel Neutralizing Monoclonal Antibody Against Ebola Virus GP.

    PubMed

    Reynard, Olivier; Volchkov, Viktor E

    2015-10-01

    Ebola virus is the etiological agent of a severe hemorrhagic fever with a high mortality rate. As the only protein exposed on the surface of viral particles, the spike glycoprotein GP is the unique target for neutralizing monoclonal antibodies. In this study, we demonstrate the strong neutralization capacity of the monoclonal antibody #3327 and characterize its activity. GP residues that are required for recognition and neutralization were found to be located both in the internal fusion loop and in the receptor-binding domain. Analysis of Ebola virus entry in the presence of #3327 allows us to hypothesize that this antibody binds to the virus particle before internalization and endosomal processing of GP and likely prevents the final viral fusion step. Importantly, #3327 is able to block entry of virions bearing GP that contain the Q508 escape mutation common to a number of virus-neutralizing antibodies, and therefore provides future perspectives for treatment strategies against Ebola virus infection.

  2. Antigenic analysis of divergent genotypes human Enterovirus 71 viruses by a panel of neutralizing monoclonal antibodies: current genotyping of EV71 does not reflect their antigenicity.

    PubMed

    Chen, Yixin; Li, Chuan; He, Delei; Cheng, Tong; Ge, Shengxiang; Shih, James Wai-Kuo; Zhao, Qinjian; Chen, Pei-Jer; Zhang, Jun; Xia, Ningshao

    2013-01-01

    In recent year, Enterovirus 71 (EV71)-associated hand, foot and mouth disease (HFMD) has become an important public health issue in China. EV71 has been classified into genotypes A, B1-B5 and C1-C5. With such genetic diversity, whether the convalescent or recovery antibody responses can cross-protect infections from other genotypes remains a question. Understanding of the antigenicity of such diverse genetic EV71 isolates is crucial for the EV71 vaccine development. Here, a total of 186 clones anti-EV71 MAbs was generated and characterized with Western blot and cell-based neutralization assay. Forty neutralizing anti-EV71 MAbs were further used to analyze the antigenic properties of 18 recent EV71 isolates representing seven genotypes in neutralization assay. We found that most neutralizing anti-EV71 MAbs are specific to conformational epitopes. We also classified the 40 neutralizing anti-EV71 MAbs into two classes according to their reactivity patterns with 18 EV71 isolates. Class I MAb can neutralize all isolates, suggesting conserved epitopes are present among EV71. Class II MAb includes four subclasses (IIa-IId) and neutralizes only subgroups of EV71 strains. Conversely, 18 EV71 strains were grouped into antigenic types 1 and four antigenic subtypes (2.1-2.4). These results suggest that the current genotyping of EV71 does not reflect their antigenicity which may be important in the selection of EV71 vaccine strains. This panel of neutralizing anti-EV71 MAbs may be useful for the recognition of emerging antigenic variants of EV71 and vaccine development.

  3. Tautomerism in neutral histidine.

    PubMed

    Bermúdez, Celina; Mata, Santiago; Cabezas, Carlos; Alonso, José L

    2014-10-01

    Histidine is an important natural amino acid, involved in many relevant biological processes, which, because of its physical properties, proved difficult to characterize experimentally in its neutral form. In this work, neutral histidine has been generated in the gas phase by laser ablation of solid samples and its N(ε)H tautomeric form unraveled through its rotational spectrum. The quadrupole hyperfine structure, arising from the existing three (14)N nuclei, constituted a site-specifically probe for revealing the tautomeric form as well as the side chain configuration of this proteogenic amino acid.

  4. Dengue virus infection-enhancing and neutralizing antibody balance in children of the Philippines and Indonesia.

    PubMed

    Yamanaka, Atsushi; Tabuchi, Yuko; Mulyatno, Kris C; Susilowati, Helen; Hendrianto, Eryk; Soegijanto, Soegeng; Konishi, Eiji

    2012-11-01

    Dengue fever and dengue hemorrhagic fever are important diseases worldwide. Although antibody-dependent enhancement of infection has been proposed as a mechanism for increased disease severity, enhancing antibodies in endemic people have not been thoroughly investigated. Recently, we established a serological assay system to measure the balance of enhancing and neutralizing activities, which provides useful information for estimating in vivo antibody status. We measured the balance of these activities against four dengue virus (DENV) types in endemic populations, and analyzed the proportion of sera containing enhancing and neutralizing antibodies. Predominantly healthy Filipino children were used for analysis, although a population of Indonesian children was also investigated. In the Filipino population, the highest proportion of neutralizing activities was shown against DENV2, followed by DENV1. A greater proportion of sera exhibited enhancing rather than neutralizing antibodies against other virus types. Neutralizing activities were complement-dependent, while enhancing activities were complement-independent. The Indonesian population showed a similar dengue antibody status. Our results indicate that a relatively high proportion of endemic children possessed complement-independent enhancing antibodies against some DENV types.

  5. Neutral buoyancy evaluation of technologies for space station external operations. [EVA weightlessness simulation

    NASA Technical Reports Server (NTRS)

    Akin, D. L.; Bowden, M. L.; Spofford, J. R.

    1984-01-01

    In order to perform a complete systems analysis for almost any large space program, it is vital to have a thorough understanding of human capabilities in extravehicular activity (EVA). The present investigation is concerned with the most significant results from the MIT Space Systems Lab's neutral buoyancy tests. An evaluation of neutral buoyancy is considered along with the tested structures, aspects of learning, productivity, time and motion analysis, and assembly loads. Attention is given to EVA assembly with a manned maneuvering unit, teleoperated structural assembly, an integrated control station, a beam assembly teleoperator, and space station proximity operations.

  6. Molecular events involved in up-regulating human Na+-independent neutral amino acid transporter LAT1 during T-cell activation.

    PubMed

    Nii, T; Segawa, H; Taketani, Y; Tani, Y; Ohkido, M; Kishida, S; Ito, M; Endou, H; Kanai, Y; Takeda, E; Miyamoto Ki

    2001-09-15

    We investigated the regulation of system-L amino acid transporter (LAT1) during T-cell activation. In quiescent T-cells, L-leucine transport is mediated mainly by the system-L amino acid transport system and is increased significantly during T-cell activation by PMA and ionomycin. In quiescent T-cells, the LAT1 protein was heterocomplexed with 4F2 heavy chain (4F2hc) in the plasma membrane. During T-cell activation, the amounts of 4F2hc and LAT1 heterocomplex were significantly elevated compared with those in quiescent T-cells. In addition, by Northern-blot analysis, these increments were found to be due to elevated levels of LAT1 and 4F2hc mRNA. Transient expression of constructs comprising various LAT1 gene promoter fragments, which contained all three of the GC boxes, was sufficient for promoting luciferase expression in Jurkat T-cells, but the promoter of the LAT1 gene did not respond to PMA and ionomycin. Similar observations were observed in the human 4F2hc gene promoter. In nuclear run-on assay, the LAT1 and 4F2hc genes were actively transcribed even in quiescent T-cells, but the low levels of both transcripts were shown to be the result of a block to transcription elongation within the exon 1 intron 1 regions. These findings indicated that a removal of the block to mRNA elongation stimulates the induction of system-L amino acid transporter gene transcripts (LAT1 and 4F2hc) in activated T-cells. PMID:11535130

  7. CO2-Neutral Fuels

    NASA Astrophysics Data System (ADS)

    Goede, Adelbert; van de Sanden, Richard

    2016-06-01

    Mimicking the biogeochemical cycle of System Earth, synthetic hydrocarbon fuels are produced from recycled CO2 and H2O powered by renewable energy. Recapturing CO2 after use closes the carbon cycle, rendering the fuel cycle CO2 neutral. Non-equilibrium molecular CO2 vibrations are key to high energy efficiency.

  8. Bleach Neutralizes Mold Allergens

    ERIC Educational Resources Information Center

    Science Teacher, 2005

    2005-01-01

    Researchers at National Jewish Medical and Research Center have demonstrated that dilute bleach not only kills common household mold, but may also neutralize the mold allergens that cause most mold-related health complaints. The study, published in the Journal of Allergy and Clinical Immunology, is the first to test the effect on allergic…

  9. Recombinant Sheep Pox Virus Proteins Elicit Neutralizing Antibodies

    PubMed Central

    Chervyakova, Olga V.; Zaitsev, Valentin L.; Iskakov, Bulat K.; Tailakova, Elmira T.; Strochkov, Vitaliy M.; Sultankulova, Kulyaisan T.; Sandybayev, Nurlan T.; Stanbekova, Gulshan E.; Beisenov, Daniyar K.; Abduraimov, Yergali O.; Mambetaliyev, Muratbay; Sansyzbay, Abylay R.; Kovalskaya, Natalia Y.; Nemchinov, Lev. G.; Hammond, Rosemarie W.

    2016-01-01

    The aim of this work was to evaluate the immunogenicity and neutralizing activity of sheep pox virus (SPPV; genus Capripoxvirus, family Poxviridae) structural proteins as candidate subunit vaccines to control sheep pox disease. SPPV structural proteins were identified by sequence homology with proteins of vaccinia virus (VACV) strain Copenhagen. Four SPPV proteins (SPPV-ORF 060, SPPV-ORF 095, SPPV-ORF 117, and SPPV-ORF 122), orthologs of immunodominant L1, A4, A27, and A33 VACV proteins, respectively, were produced in Escherichia coli. Western blot analysis revealed the antigenic and immunogenic properties of SPPV-060, SPPV-095, SPPV-117 and SPPV-122 proteins when injected with adjuvant into experimental rabbits. Virus-neutralizing activity against SPPV in lamb kidney cell culture was detected for polyclonal antisera raised to SPPV-060, SPPV-117, and SPPV-122 proteins. To our knowledge, this is the first report demonstrating the virus-neutralizing activities of antisera raised to SPPV-060, SPPV-117, and SPPV-122 proteins. PMID:27338444

  10. Comparative Sensitivity Analysis of Muscle Activation Dynamics.

    PubMed

    Rockenfeller, Robert; Günther, Michael; Schmitt, Syn; Götz, Thomas

    2015-01-01

    We mathematically compared two models of mammalian striated muscle activation dynamics proposed by Hatze and Zajac. Both models are representative for a broad variety of biomechanical models formulated as ordinary differential equations (ODEs). These models incorporate parameters that directly represent known physiological properties. Other parameters have been introduced to reproduce empirical observations. We used sensitivity analysis to investigate the influence of model parameters on the ODE solutions. In addition, we expanded an existing approach to treating initial conditions as parameters and to calculating second-order sensitivities. Furthermore, we used a global sensitivity analysis approach to include finite ranges of parameter values. Hence, a theoretician striving for model reduction could use the method for identifying particularly low sensitivities to detect superfluous parameters. An experimenter could use it for identifying particularly high sensitivities to improve parameter estimation. Hatze's nonlinear model incorporates some parameters to which activation dynamics is clearly more sensitive than to any parameter in Zajac's linear model. Other than Zajac's model, Hatze's model can, however, reproduce measured shifts in optimal muscle length with varied muscle activity. Accordingly we extracted a specific parameter set for Hatze's model that combines best with a particular muscle force-length relation. PMID:26417379

  11. Neutron activation analysis in archaeological chemistry

    SciTech Connect

    Harbottle, G.

    1987-01-01

    Neutron activation analysis has proven to be a convenient way of performing the chemical analysis of archaeologically-excavated artifacts and materials. It is fast and does not require tedious laboratory operations. It is multielement, sensitive, and can be made nondestructive. Neutron activation analysis in its instrumental form, i.e., involving no chemical separation, is ideally suited to automation and conveniently takes the first step in data flow patterns that are appropriate for many taxonomic and statistical operations. The future will doubtless see improvements in the practice of NAA in general, but in connection with archaeological science the greatest change will be the filling, interchange and widespread use of data banks based on compilations of analytical data. Since provenience-oriented data banks deal with materials (obsidian, ceramics, metals, semiprecious stones, building materials and sculptural media) that participated in trade networks, the analytical data is certain to be of interest to a rather broad group of archaeologists. It is to meet the needs of the whole archaeological community that archaeological chemistry must now turn.

  12. NADP-dependent dehydrogenases in rat liver parenchyma. III. The description of a liponeogenic area on the basis of histochemically demonstrated enzyme activities and the neutral fat content during fasting and refeeding.

    PubMed

    Rieder, H

    1981-01-01

    The activities of glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase(6PGDH), malic enzyme (ME) and isocitrate dehydrogenase (ICDh) were investigated with optimized histochemical methods (Rieder it al 1978), and the activity of 3-hydroxybutyrate dehydrogenase (3HBDH) and neutral fat content with conventional techniques in the liver of male rats under the following experimental dietary conditions: (A) Fasting for 0, 12 and 84h; (B) 84-h fasting followed by refeeding with a low-fat, high-carbohydrate diet for 6 h and for 2, 3, 5, 7, 11 and 14 nights; (C) refeeding with standard diet for 5 nights; (D) low-fat high-carbohydrate diet for 7 an 14 nights. The activities of G6PDH, 6PGDH and ME decreased slightly during fasting primarily in zone 1 and increased dramatically on refeeding with a low-fat, high-carbohydrate diet. This activity increase was confined mainly to zone 3 during the first 3 days and was accompanied by a deposition of neutral fats that began in zone 3 and progressed to zone 1. Neutral for accumulation was maximal after 3 nights, with a uniform accumulation of large droplets in all the hepatocytes; this was followed by a release that started in zone 3 and proceeded in a periportal direction. On the other hand, G6PDH, 6PGDH and ME attained their maximum activities after 5 amd 7 nights of low-fat diet, the activities being nearly homogeneously distributed over the liver acinus in a few cases. Subsequently the activities fill mainly in zone 1, causing the activity patterns and levels to approach those of the animals in group (D). In contrast to this, the activity of ICDH increased during fasting principally in zone 1, so that the otherwise steep activity gradient in favor of zone 3 lessened. Refeeding led at first to a fall of activity below the initial value, but later the normal distribution pattern was restored. The activity of 3HBDH showed a behavior similar to that of ICDH. The findings are discussed with reference to the

  13. Analysis of DOE international environmental management activities

    SciTech Connect

    Ragaini, R.C.

    1995-09-01

    The Department of Energy`s (DOE) Strategic Plan (April 1994) states that DOE`s long-term vision includes world leadership in environmental restoration and waste management activities. The activities of the DOE Office of Environmental Management (EM) can play a key role in DOE`s goals of maintaining U.S. global competitiveness and ensuring the continuation of a world class science and technology community. DOE`s interest in attaining these goals stems partly from its participation in organizations like the Trade Policy Coordinating Committee (TPCC), with its National Environmental Export Promotion Strategy, which seeks to strengthen U.S. competitiveness and the building of public-private partnerships as part of U.S. industrial policy. The International Interactions Field Office task will build a communication network which will facilitate the efficient and effective communication between DOE Headquarters, Field Offices, and contractors. Under this network, Headquarters will provide the Field Offices with information on the Administration`s policies and activities (such as the DOE Strategic Plan), interagency activities, as well as relevant information from other field offices. Lawrence Livermore National Laboratory (LLNL) will, in turn, provide Headquarters with information on various international activities which, when appropriate, will be included in reports to groups like the TPCC and the EM Focus Areas. This task provides for the collection, review, and analysis of information on the more significant international environmental restoration and waste management initiatives and activities which have been used or are being considered at LLNL. Information gathering will focus on efforts and accomplishments in meeting the challenges of providing timely and cost effective cleanup of its environmentally damaged sites and facilities, especially through international technical exchanges and/or the implementation of foreign-development technologies.

  14. Scalable histopathological image analysis via active learning.

    PubMed

    Zhu, Yan; Zhang, Shaoting; Liu, Wei; Metaxas, Dimitris N

    2014-01-01

    Training an effective and scalable system for medical image analysis usually requires a large amount of labeled data, which incurs a tremendous annotation burden for pathologists. Recent progress in active learning can alleviate this issue, leading to a great reduction on the labeling cost without sacrificing the predicting accuracy too much. However, most existing active learning methods disregard the "structured information" that may exist in medical images (e.g., data from individual patients), and make a simplifying assumption that unlabeled data is independently and identically distributed. Both may not be suitable for real-world medical images. In this paper, we propose a novel batch-mode active learning method which explores and leverages such structured information in annotations of medical images to enforce diversity among the selected data, therefore maximizing the information gain. We formulate the active learning problem as an adaptive submodular function maximization problem subject to a partition matroid constraint, and further present an efficient greedy algorithm to achieve a good solution with a theoretically proven bound. We demonstrate the efficacy of our algorithm on thousands of histopathological images of breast microscopic tissues. PMID:25320821

  15. Electrodermal activity analysis during affective haptic elicitation.

    PubMed

    Greco, Alberto; Valenza, Gaetano; Nardelli, Mimma; Bianchi, Matteo; Lanata, Antonio; Scilingo, Enzo Pasquale

    2015-08-01

    This paper investigates how the autonomic nervous system dynamics, quantified through the analysis of the electrodermal activity (EDA), is modulated according to affective haptic stimuli. Specifically, a haptic display able to convey caress-like stimuli is presented to 32 healthy subjects (16 female). Each stimulus is changed according to six combinations of three velocities and two forces levels of two motors stretching a strip of fabric. Subjects were also asked to score each stimulus in terms of arousal (high/low activation) and valence (pleasant/unpleasant), in agreement with the circumplex model of affect. EDA was processed using a deconvolutive method, separating tonic and phasic components. A statistical analysis was performed in order to identify significant differences in EDA features among force and velocity levels, as well as in their valence and arousal scores. Results show that the simulated caress induced by the haptic display significantly affects the EDA. In detail, the phasic component seems to be inversely related to the valence score. This finding is new and promising, since it can be used, e.g., as an additional cue for haptics design. PMID:26737605

  16. NMR-monitored titration of acid-stress bacterial chaperone HdeA reveals that Asp and Glu charge neutralization produces a loosened dimer structure in preparation for protein unfolding and chaperone activation.

    PubMed

    Garrison, McKinzie A; Crowhurst, Karin A

    2014-02-01

    HdeA is a periplasmic chaperone found in several gram-negative pathogenic bacteria that are linked to millions of cases of dysentery per year worldwide. After the protein becomes activated at low pH, it can bind to other periplasmic proteins, protecting them from aggregation when the bacteria travel through the stomach on their way to colonize the intestines. It has been argued that one of the major driving forces for HdeA activation is the protonation of aspartate and glutamate side chains. The goal for this study, therefore, was to investigate, at the atomic level, the structural impact of this charge neutralization on HdeA during the transition from near-neutral conditions to pH 3.0, in preparation for unfolding and activation of its chaperone capabilities. NMR spectroscopy was used to measure pKa values of Asp and Glu residues and monitor chemical shift changes. Measurements of R2/R1 ratios from relaxation experiments confirm that the protein maintains its dimer structure between pH 6.0 and 3.0. However, calculated correlation times and changes in amide protection from hydrogen/deuterium exchange experiments provide evidence for a loosening of the tertiary and quaternary structures of HdeA; in particular, the data indicate that the dimer structure becomes progressively weakened as the pH decreases. Taken together, these results provide insight into the process by which HdeA is primed to unfold and carry out its chaperone duties below pH 3.0, and it also demonstrates that neutralization of aspartate and glutamate residues is not likely to be the sole trigger for HdeA dissociation and unfolding.

  17. Antihypertensive neutral lipid

    DOEpatents

    Snyder, F.L.; Blank, M.L.

    1984-10-26

    The invention relates to the discovery of a class of neutral acetylated either-linked glycerolipids having the capacity to lower blood presure in warm-blooded animals. This physiological effect is structure sensitive requiring a long chain alkyl group at the sn-1 position and a short carbon chain acyl group (acetyl or propionyl) at the sn-2 position, and a hydroxyl group at the sn-3 position.

  18. Antihypertensive neutral lipid

    DOEpatents

    Snyder, Fred L.; Blank, Merle L.

    1986-01-01

    The invention relates to the discovery of a class of neutral acetylated ether-linked glycerolipids having the capacity to lower blood pressure in warm-blooded animals. This physiological effect is structure sensitive requiring a long chain alkyl group at the sn-1 position and a short carbon chain acyl group (acetyl or propionyl) at the sn-2 position, and a hydroxyl group at the sn-3 position.

  19. Neutral atom traps.

    SciTech Connect

    Pack, Michael Vern

    2008-12-01

    This report describes progress in designing a neutral atom trap capable of trapping sub millikelvin atom in a magnetic trap and shuttling the atoms across the atom chip from a collection area to an optical cavity. The numerical simulation and atom chip design are discussed. Also, discussed are preliminary calculations of quantum noise sources in Kerr nonlinear optics measurements based on electromagnetically induced transparency. These types of measurements may be important for quantum nondemolition measurements at the few photon limit.

  20. Exercise Equipment: Neutral Buoyancy

    NASA Technical Reports Server (NTRS)

    Shackelford, Linda; Valle, Paul

    2016-01-01

    Load Bearing Equipment for Neutral Buoyancy (LBE-NB) is an exercise frame that holds two exercising subjects in position as they apply counter forces to each other for lower extremity and spine loading resistance exercises. Resistance exercise prevents bone loss on ISS, but the ISS equipment is too massive for use in exploration craft. Integrating the human into the load directing, load generating, and motion control functions of the exercise equipment generates safe exercise loads with less equipment mass and volume.

  1. Results on intense beam focusing and neutralization from the neutralized beam experiment

    SciTech Connect

    Roy, P.K.; Yu, S.S.; Eylon, S.; Henestroza, E.; Anders, A.; Bieniosek, F.M.; Greenway, W.G.; Logan, B.G.; Waldron, W.L.; Vanecek, D.L.; Welch, D.R.; Rose, D.V.; Davidson, R.C.; Efthimion, P.C.; Gilson, E.P.; Sefkow, A.B.; Sharp, W.M.

    2003-10-31

    We have demonstrated experimental techniques to provide active neutralization for space-charge dominated beams as well as to prevent uncontrolled ion beam neutralization by stray electrons. Neutralization is provided by a localized plasma injected from a cathode arc source. Unwanted secondary electrons produced at the wall by halo particle impact are suppressed using a radial mesh liner that is positively biased inside a beam drift tube. We present measurements of current transmission, beam spot size as a function of axial position, beam energy and plasma source conditions. Detailed comparisons with theory are also presented.

  2. Thermal neutral format based on the step technology

    NASA Technical Reports Server (NTRS)

    Almazan, P. Planas; Legal, J. L.

    1995-01-01

    The exchange of models is one of the most serious problems currently encountered in the practice of spacecraft thermal analysis. Essentially, the problem originates in the diversity of computing environments that are used across different sites, and the consequent proliferation of native tool formats. Furthermore, increasing pressure to reduce the development's life cycle time has originated a growing interest in the so-called spacecraft concurrent engineering. In this context, the realization of the interdependencies between different disciplines and the proper communication between them become critical issues. The use of a neutral format represents a step forward in addressing these problems. Such a means of communication is adopted by consensus. A neutral format is not directly tied to any specific tool and it is kept under stringent change control. Currently, most of the groups promoting exchange formats are contributing with their experience to STEP, the Standard for Exchange of Product Model Data, which is being developed under the auspices of the International Standards Organization (ISO 10303). This paper presents the different efforts made in Europe to provide the spacecraft thermal analysis community with a Thermal Neutral Format (TNF) based on STEP. Following an introduction with some background information, the paper presents the characteristics of the STEP standard. Later, the first efforts to produce a STEP Spacecraft Thermal Application Protocol are described. Finally, the paper presents the currently harmonized European activities that follow up and extend earlier work on the area.

  3. Neutral particle beam intensity controller

    DOEpatents

    Dagenhart, W.K.

    1984-05-29

    The neutral beam intensity controller is based on selected magnetic defocusing of the ion beam prior to neutralization. The defocused portion of the beam is dumped onto a beam dump disposed perpendicular to the beam axis. Selective defocusing is accomplished by means of a magnetic field generator disposed about the neutralizer so that the field is transverse to the beam axis. The magnetic field intensity is varied to provide the selected partial beam defocusing of the ions prior to neutralization. The desired focused neutral beam portion passes along the beam path through a defining aperture in the beam dump, thereby controlling the desired fraction of neutral particles transmitted to a utilization device without altering the kinetic energy level of the desired neutral particle fraction. By proper selection of the magnetic field intensity, virtually zero through 100% intensity control of the neutral beam is achieved.

  4. Rabbits immunized with Epstein-Barr virus gH/gL or gB recombinant proteins elicit higher serum virus neutralizing activity than gp350.

    PubMed

    Cui, Xinle; Cao, Zhouhong; Chen, Quanyi; Arjunaraja, Swadhinya; Snow, Andrew L; Snapper, Clifford M

    2016-07-25

    Epstein-Barr virus (EBV) is the primary cause of infectious mononucleosis and has been strongly implicated in the etiology of multiple epithelial and lymphoid cancers, such as nasopharyngeal carcinoma, gastric carcinoma, Hodgkin lymphoma, Burkitt lymphoma, non-Hodgkin lymphoma and post-transplant lymphoproliferative disorder. There is currently no licensed prophylactic vaccine for EBV. Most efforts to develop prophylactic vaccines have focused on EBV gp350, which binds to CD21/CD35 to gain entry into B cells, and is a major target of serum neutralizing antibody in EBV seropositive humans. However, a recombinant monomeric gp350 protein failed to prevent EBV infection in a phase II clinical trial. Thus, alternative or additional target antigens may be necessary for a successful prophylactic vaccine. EBV gH/gL and gB proteins coordinately mediate EBV fusion and entry into B cells and epithelial cells, strongly suggesting that vaccination with these proteins might elicit antibodies that will prevent EBV infection. We produced recombinant trimeric and monomeric EBV gH/gL heterodimeric proteins and a trimeric EBV gB protein, in addition to tetrameric and monomeric gp350(1-470) proteins, in Chinese hamster ovary cells. We demonstrated that vaccination of rabbits with trimeric and monomeric gH/gL, trimeric gB, and tetrameric gp350(1-470) induced serum EBV-neutralizing titers, using cultured human B cells, that were >100-fold, 20-fold, 18-fold, and 4-fold higher, respectively, than monomeric gp350(1-470). These data strongly suggest a role for testing EBV gH/gL and EBV gB in a future prophylactic vaccine to prevent EBV infection of B cells, as well as epithelial cells.

  5. Exchanging physically effective neutral detergent fiber does not affect chewing activity and performance of late-lactation dairy cows fed corn and sugarcane silages.

    PubMed

    Sá Neto, A; Bispo, A W; Junges, D; Bercht, A K; Zopollatto, M; Daniel, J L P; Nussio, L G

    2014-11-01

    The objective of this study was to determine whether replacing the physically effective neutral detergent fiber (peNDF) of corn silage with sugarcane silage peNDF would affect performance in dairy cows. Twenty-four late-lactation Holstein cows were assigned to eight 3 × 3 Latin squares with 21-d periods. The dietary treatments were (1) 25% peNDF of corn silage, (2) 25% peNDF of sugarcane silage, and (3) 12.5% peNDF of corn silage + 12.5% peNDF of sugarcane silage. The physical effectiveness factors (pef) were assumed to be 1 for corn silage and 1.2 for sugarcane silage, as measured previously by bioassay. Thus, peNDF was calculated as neutral detergent fiber (NDF) × pef. The concentrate ingredients were finely ground corn, soybean meal, pelleted citrus pulp, and mineral-vitamin premix. Dry matter intake (22.5 ± 0.63 kg/d), 3.5% fat-corrected milk yield (28.8 ± 1.13 kg/d), milk composition (fat, protein, lactose, urea, casein, free fatty acids, and somatic cell count), and blood metabolites (glucose, insulin, and nonesterified fatty acids) were unaffected by the treatments. The time spent eating, ruminating, or chewing was also similar among the diets, as was particle-sorting behavior. By contrast, chewing per kilogram of forage NDF intake was higher for the sugarcane silage (137 min/kg) than the corn silage diet (116 min/kg), indicating the greater physical effectiveness of sugarcane fiber. Based on chewing behavior (min/d), the estimated pef of sugarcane silage NDF were 1.28 in the corn silage plus sugarcane silage diet and 1.29 in the sugarcane silage diet. Formulating dairy rations of equal peNDF content allows similar performance if corn and sugarcane silages are exchanged.

  6. Cross-Reactivity of Neutralizing Antibodies among Malignant Catarrhal Fever Viruses.

    PubMed

    Taus, Naomi S; Cunha, Cristina W; Marquard, Jana; O'Toole, Donal; Li, Hong

    2015-01-01

    Some members of the gamma herpesvirus genus Macavirus are maintained in nature as subclinical infections in well-adapted ungulate hosts. Transmission of these viruses to poorly adapted hosts, such as American bison and cattle, can result in the frequently fatal disease malignant catarrhal fever (MCF). Based on phylogenetic analysis, the MCF viruses (MCFV) cluster into two subgroups corresponding to the reservoir hosts' subfamilies: Alcelaphinae/Hippotraginae and Caprinae. Antibody cross-reactivity among MCFVs has been demonstrated using techniques such as enzyme linked immunosorbent and immunofluorescence assays. However, minimal information is available as to whether virus neutralizing antibodies generated against one MCFV cross react with other members of the genus. This study tested the neutralizing activity of serum and plasma from select MCFV-infected reservoir hosts against alcelaphine herpesvirus 1 (AlHV-1) and ovine herpesvirus 2 (OvHV-2). Neutralizing antibody activity against AlHV-1 was detected in samples from infected hosts in the Alcelaphinae and Hippotraginae subfamilies, but not from hosts in the Caprinae subfamily. OvHV-2 neutralizing activity was demonstrated in samples from goats (Caprinae) but not from wildebeest (Alcelaphinae). These results show that neutralizing antibody cross reactivity is present to MCFVs within a virus subgroup but not between subgroups. This information is important for diagnosing infection with MCFVs and in the development of vaccines against MCF. PMID:26658281

  7. Neutrality between Government and Religion.

    ERIC Educational Resources Information Center

    Mawdsley, Ralph D.

    1996-01-01

    The overall guiding principle of neutrality between government and religion masks a tension that exists between free exercise of religion and establishment of religion. Reviews the development and current status of "Lemon" as a test for neutrality; proposes a new test for neutrality, evenhandedness, that is common to both the Free Exercise and…

  8. Antibody Response to Hypervariable Region 1 Interferes with Broadly Neutralizing Antibodies to Hepatitis C Virus

    PubMed Central

    Keck, Zhen-yong; Girard-Blanc, Christine; Wang, Wenyan; Lau, Patrick; Zuiani, Adam; Rey, Felix A.; Krey, Thomas; Diamond, Michael S.

    2015-01-01

    ABSTRACT Hypervariable region 1 (HVR1) (amino acids [aa] 384 to 410) on the E2 glycoprotein of hepatitis C virus contributes to persistent infection by evolving escape mutations that attenuate binding of inhibitory antibodies and by blocking access of broadly neutralizing antibodies to their epitopes. A third proposed mechanism of immune antagonism is that poorly neutralizing antibodies binding to HVR1 interfere with binding of other superior neutralizing antibodies. Epitope mapping of human monoclonal antibodies (HMAbs) that bind to an adjacent, conserved domain on E2 encompassing aa 412 to 423 revealed two subsets, designated HC33 HMAbs. While both subsets have contact residues within aa 412 to 423, alanine-scanning mutagenesis suggested that one subset, which includes HC33.8, has an additional contact residue within HVR1. To test for interference of anti-HVR1 antibodies with binding of antibodies to aa 412 to 423 and other E2 determinants recognized by broadly neutralizing HMAbs, two murine MAbs against HVR1 (H77.16) and aa 412 to 423 (H77.39) were studied. As expected, H77.39 inhibited the binding of all HC33 HMAbs. Unexpectedly, H77.16 also inhibited the binding of both subsets of HC33 HMAbs. This inhibition also was observed against other broadly neutralizing HMAbs to epitopes outside aa 412 to 423. Combination antibody neutralization studies by the median-effect analysis method with H77.16 and broadly reactive HMAbs revealed antagonism between these antibodies. Structural studies demonstrated conformational flexibility in this antigenic region, which supports the possibility of anti-HVR1 antibodies hindering the binding of broadly neutralizing MAbs. These findings support the hypothesis that anti-HVR1 antibodies can interfere with a protective humoral response against HCV infection. IMPORTANCE HVR1 contributes to persistent infection by evolving mutations that escape from neutralizing antibodies to HVR1 and by shielding broadly neutralizing antibodies from

  9. Analysis of physical activities in Taekwondo Pumsae.

    PubMed

    Lee, Sang-Bock; Cha, Eun-Jong; Lee, Tae-Soo

    2008-01-01

    Exercise is very important element for successful aging. Among many sports events, Korea is the suzerain of Taekwondo. When competing (Taekwondo Free Fighting) after learning Poomse as basic movements and inuring them, people compete with movements depending on situation. Among Poomses of Taekwondo, Taegeuk Poomse consists of very basic movements from 1 Jang to 8 Jang and they are for inuring to body. In order to prescribe Taegeuk Jang, which is the basic movement of Taekwondo that Korea is the suzerain, as an exercise for successful aging, it is necessary to analyze physical activity level of each Taegeuk Jang (From 1 Jang through 8 Jang) and suggest the same. Therefore, in this study, I analyzed physical activity level of each Jang of Taegeuk Poomse by attaching Armband made by Body Media Company on brachia and legs below knee of Taekwondo trainees. The result of the analysis of the whole momentum from Taegeuk 1 Jang to 8 Jang is as follows: First, the average absolute value of acceleration variation of vertical direction signal (L-MAD): 5.15. Second, the average absolute value of acceleration variation of horizontal direction signal (T-MAD): 3.44. Finally, the average of calorie consumption per minute (AEE/Min): 5.06 Cal. The obtained result corresponds to proper exercise condition for successful aging and it can be utilized as data for exercise prescription for the young and the old.

  10. Active polarimeter optical system laser hazard analysis.

    SciTech Connect

    Augustoni, Arnold L.

    2005-07-01

    A laser hazard analysis was performed for the SNL Active Polarimeter Optical System based on the ANSI Standard Z136.1-2000, American National Standard for Safe Use of Lasers and the ANSI Standard Z136.6-2000, American National Standard for Safe Use of Lasers Outdoors. The Active Polarimeter Optical System (APOS) uses a pulsed, near-infrared, chromium doped lithium strontium aluminum fluoride (Cr:LiSAF) crystal laser in conjunction with a holographic diffuser and lens to illuminate a scene of interest. The APOS is intended for outdoor operations. The system is mounted on a height adjustable platform (6 feet to 40 feet) and sits atop a tripod that points the beam downward. The beam can be pointed from nadir to as much as 60 degrees off of nadir producing an illuminating spot geometry that can vary from circular (at nadir) to elliptical in shape (off of nadir). The JP Innovations crystal Cr:LiSAF laser parameters are presented in section II. The illuminating laser spot size is variable and can be adjusted by adjusting the separation distance between the lens and the holographic diffuser. The system is adjusted while platform is at the lowest level. The laser spot is adjusted for a particular spot size at a particular distance (elevation) from the laser by adjusting the separation distance (d{sub diffuser}) to predetermined values. The downward pointing angle is also adjusted before the platform is raised to the selected operation elevation.

  11. Analysis of retrotransposon activity in plants.

    PubMed

    Defraia, Christopher; Slotkin, R Keith

    2014-01-01

    Retrotransposons are transposable elements that duplicate themselves by converting their transcribed RNA genome into cDNA, which is then integrated back into the genome. Retrotransposons can be divided into two major classes based on their mechanism of transposition and the presence or absence of long terminal repeats (LTRs). In contrast to mammalian genomes, in which non-LTR retrotransposons have proliferated, plant genomes show evolutionary evidence of an explosion in LTR retrotransposon copy number. These retrotransposons can comprise a large fraction of the genome (75 % in maize). Although often viewed as molecular parasites, retrotransposons have been shown to influence neighboring gene expression and play a structural and potential regulatory role in the centromere. To prevent retrotransposon activity, eukaryotic cells have evolved overlapping mechanisms to repress transposition. Plants are an excellent system for studying the mechanisms of LTR retrotransposon inhibition such as DNA methylation and small RNA-mediated degradation of retrotransposon transcripts. However, analysis of these multi-copy, mobile elements is considerably more difficult than analysis of single-copy genes located in stable regions of the genome. In this chapter we outline methods for analyzing the progress of LTR retrotransposons through their replication cycle in plants. We describe a mixture of traditional molecular biology experiments, such as Southern, Northern, and Western blotting, in addition to nontraditional techniques designed to take advantage of the specific mechanism of LTR retrotransposition.

  12. Neutral Particle Analyzer Measurements of Ion Behavior in NSTX

    SciTech Connect

    S.S. Medley; R.E. Bell; D.S. Darrow; A.L. Roquemore

    2002-02-06

    Initial results obtained with the Neutral Particle Analyzer (NPA) diagnostic on the National Spherical Torus Experiment (NSTX) are presented. Magnetohydrodynamic activity and reconnection events cause depletion of the deuterium energetic ion distribution created by neutral-beam injection. Adding High Harmonic Fast Wave Heating to neutral-beam-heated discharges results in the generation of an energetic ion tail above the beam injection energy. NPA measurements of the residual hydrogen ion temperature are in good agreement with those from recombination spectroscopy.

  13. Pulsed field sample neutralization

    DOEpatents

    Appelhans, Anthony D.; Dahl, David A.; Delmore, James E.

    1990-01-01

    An apparatus and method for alternating voltage and for varying the rate of extraction during the extraction of secondary particles, resulting in periods when either positive ions, or negative ions and electrons are extracted at varying rates. Using voltage with alternating charge during successive periods to extract particles from materials which accumulate charge opposite that being extracted causes accumulation of surface charge of opposite sign. Charge accumulation can then be adjusted to a ratio which maintains a balance of positive and negative charge emission, thus maintaining the charge neutrality of the sample.

  14. Coherent and neutral pion production results from MINERνA

    SciTech Connect

    Palomino, J. L.; Higuera, A.

    2015-05-15

    MINERνA is a neutrino-nucleus scattering experiment employing multiple nuclear targets. The experiment is studying neutral pion production due to coherent, resonant and deep-inelastic processes, from both charged current and neutral current reactions. Neutral pions are detected through their two photon decay and the resultant electromagnetic showers. We will describe the analysis for the cross sections of inclusive and exclusive processes.

  15. Assessing physical activity intensity by video analysis.

    PubMed

    Silva, P; Santiago, C; Reis, L P; Sousa, A; Mota, J; Welk, G

    2015-05-01

    Assessing physical activity (PA) is a challenging task and many different approaches have been proposed. Direct observation (DO) techniques can objectively code both the behavior and the context in which it occurred, however, they have significant limitations such as the cost and burden associated with collecting and processing data. Therefore, this study evaluated the utility of an automated video analysis system (CAM) designed to record and discriminate the intensity of PA using a subject tracking methodology. The relative utility of the CAM system and DO were compared with criterion data from an objective accelerometry-based device (Actigraph GT3X+). Eight 10 year old children (three girls and five boys) wore the GT3X+ during a standard basketball session. PA was analyzed by two observers using the SOPLAY instrument and by the CAM system. The GT3X+ and the CAM were both set up to collect data at 30 Hz while the DO was performed every two minutes, with 10 s of observation for each gender. The GT3X+ was processed using cut points by Evanson and the outcome measure was the percentage of time spent in different intensities of PA. The CAM data were processed similarly using the same speed thresholds as were used in establishing the Evenson cut-off points (light: <2 mph; walking: 2-4 mph; very active: >4 mph). Similar outcomes were computed from the SOPLAY default analyses. A chi-square test was used to test differences in the percentage of time at the three intensity zones (light, walking and very active). The Yates' correction was used to prevent overestimation of statistical significance for small data. When compared with GT3X+, the CAM had better results than the SOPLAY. The chi-square test yielded the following pairwise comparisons: CAM versus GT3x+ was χ(2) (5) = 24.18, p < .001; SOPLAY2 versus GT3x+ was χ(2) (5) = 144.44, p < .001; SOPLAY1 versus GT3x+ was χ(2) (5) = 119.55, p < .001. The differences were smaller between CAM and GT3x

  16. New applications of ORNL neutral beam injectors

    SciTech Connect

    Tsai, C. C.; Peng, Yueng Kay Martin

    1998-01-01

    The injection of energetic hydrogen and deuterium atoms has been used to heat plasmas in various fusion experimental devices including tokamaks, mirrors, and stellarators. The neutral beam injection is a proven plasma heating technique for increasing plasma densities, temperatures, and pressures. For this fusion endeavor, scientists at Oak Ridge National Laboratory (ORNL) have developed multimegawatt neutral beam injectors. Various ORNL injectors have been used for studying properties of beam-heated plasmas in Oak Ridge Tokamak (ORMAK), Impurity Study Experiment-B (ISX-B), Advanced Toroidal Facility (ATF), Princeton Large Torus (PLT), and Princeton Divertor Experiment (PDX) in the United States and in Small Tight Aspect Ratio Tokamak (START) in the United Kingdom. By using a 30-cm and 100-A ion source, each ORNL neutral beam injector is capable of injecting >1.5 MW of hydrogen atoms at 50-keV for a pulse length up to 0.5 s. For increasing plasma densities and raising plasma temperatures in START, one such injector was installed and commissioned during 1995. The initial goal was to provide an injected neutral beam power of more than 0.5 MW at a beam energy of 40 keV for 20 ms. Addition of a getter pump has allowed the beam power to be raised to 1 MW at 33 keV. Recent experiments have demonstrated that neutral beam heating can play a big role in raising plasma pressures to a record volume-average beta value over 30%. ORNL neutral beam injectors have been approved for plasma heating experiments on both the TJ-II stellarator at CIEMAT, Spain, and the Mega-Amp Spherical Tokamak (MAST) at Culham Science Centre (Culham). Two proven ORNL NE injectors are being installed at the TJ-II facility. Additional ORNL beam equipment is being used to implement two 5-s NE injectors at the MAST facility. In this paper, we report and discuss the progress and plans for these neutral beam activities.

  17. Twigs on the tree of life? Neutral and selective models for integrating macroevolutionary patterns with microevolutionary processes in the analysis of asexuality.

    PubMed

    Schwander, Tanja; Crespi, Bernard J

    2009-01-01

    Neutral models characterize evolutionary or ecological patterns expected in the absence of specific causal processes, such as natural selection or ecological interactions. In this study, we describe and evaluate three neutral models that can, in principle, help to explain the apparent 'twigginess' of asexual lineages on phylogenetic trees without involving the negative consequences predicted for the absence of recombination and genetic exchange between individuals. Previously, such phylogenetic twiggyness of asexual lineages has been uncritically interpreted as evidence that asexuality is associated with elevated extinction rates and thus represents an evolutionary dead end. Our first model uses simple phylogenetic simulations to illustrate that, with sexual reproduction as the ancestral state, low transition rates to stable asexuality, or low rates of ascertained 'speciation' in asexuals, can generate twiggy distributions of asexuality, in the absence of high extinction rates for asexual lineages. The second model, developed by Janko et al. (2008), shows that a dynamic equilibrium between origins and neutral losses of asexuals can, under some conditions, generate a relatively low mean age of asexual lineages. The third model posits that the risk of extinction for asexual lineages may be higher than that of sexuals simply because asexuals inhabit higher latitudes or altitudes, and not due to effects of their reproductive systems. Such neutral models are useful in that they allow quantitative evaluation of whether empirical data, such as phylogenetic and phylogeographic patterns of sex and asexuality, indeed support the idea that asexually reproducing lineages persist over shorter evolutionary periods than sexual lineages, due to such processes as mutation accumulation, slower rates of adaptive evolution, or relatively lower levels of genetic variability.

  18. OGO-6 neutral atmospheric composition experiment

    NASA Technical Reports Server (NTRS)

    Taeusch, D. R.

    1973-01-01

    The continued analysis of data obtained from the neutral atmospheric composition experiment flown on OGO-V6 is discussed. The effort was directed toward the study of five specific areas of interest for which the OGO-V6 data were especially useful.

  19. Retinol binding protein and vitamin D associations with serum antibody isotypes, serum influenza virus-specific neutralizing activities and airway cytokine profiles.

    PubMed

    Jones, B G; Oshansky, C M; Bajracharya, R; Tang, L; Sun, Y; Wong, S S; Webby, R; Thomas, P G; Hurwitz, J L

    2016-02-01

    Vitamin A supports the induction of immunoglobulin (Ig)A responses at mucosal surfaces in mice, but much less is known about the influence of vitamins on antibody isotype expression in humans. To address this knowledge gap, we examined 46 residual blood samples from adults and children, some of whom were experiencing influenza virus infections of the respiratory tract. Assays were performed for retinol binding protein (RBP, a surrogate for vitamin A), vitamin D (a related vitamin) and antibody isotypes. Results showed that all but two tested samples exhibited RBP and/or vitamin D insufficiencies or deficiencies. Vitamin D correlated with blood IgM and IgG3, while RBP correlated with IgG4 and IgA. RBP also correlated positively with age and with influenza virus-specific antibody neutralization titres. Individuals with low blood RBP levels exhibited the highest frequencies of over-expressed cytokines and growth factors in nasal wash samples, an indication of inflamed mucosal tissues. While cause-effect relationships were not discerned, results support a hypothesis that vitamins directly influence B cell isotype expression in humans, and by so doing may help protect mucosal surfaces from respiratory viral disease. PMID:26425827

  20. A new balancing three level three dimensional space vector modulation strategy for three level neutral point clamped four leg inverter based shunt active power filter controlling by nonlinear back stepping controllers.

    PubMed

    Chebabhi, Ali; Fellah, Mohammed Karim; Kessal, Abdelhalim; Benkhoris, Mohamed F

    2016-07-01

    In this paper is proposed a new balancing three-level three dimensional space vector modulation (B3L-3DSVM) strategy which uses a redundant voltage vectors to realize precise control and high-performance for a three phase three-level four-leg neutral point clamped (NPC) inverter based Shunt Active Power Filter (SAPF) for eliminate the source currents harmonics, reduce the magnitude of neutral wire current (eliminate the zero-sequence current produced by single-phase nonlinear loads), and to compensate the reactive power in the three-phase four-wire electrical networks. This strategy is proposed in order to gate switching pulses generation, dc bus voltage capacitors balancing (conserve equal voltage of the two dc bus capacitors), and to switching frequency reduced and fixed of inverter switches in same times. A Nonlinear Back Stepping Controllers (NBSC) are used for regulated the dc bus voltage capacitors and the SAPF injected currents to robustness, stabilizing the system and to improve the response and to eliminate the overshoot and undershoot of traditional PI (Proportional-Integral). Conventional three-level three dimensional space vector modulation (C3L-3DSVM) and B3L-3DSVM are calculated and compared in terms of error between the two dc bus voltage capacitors, SAPF output voltages and THDv, THDi of source currents, magnitude of source neutral wire current, and the reactive power compensation under unbalanced single phase nonlinear loads. The success, robustness, and the effectiveness of the proposed control strategies are demonstrated through simulation using Sim Power Systems and S-Function of MATLAB/SIMULINK.

  1. A new balancing three level three dimensional space vector modulation strategy for three level neutral point clamped four leg inverter based shunt active power filter controlling by nonlinear back stepping controllers.

    PubMed

    Chebabhi, Ali; Fellah, Mohammed Karim; Kessal, Abdelhalim; Benkhoris, Mohamed F

    2016-07-01

    In this paper is proposed a new balancing three-level three dimensional space vector modulation (B3L-3DSVM) strategy which uses a redundant voltage vectors to realize precise control and high-performance for a three phase three-level four-leg neutral point clamped (NPC) inverter based Shunt Active Power Filter (SAPF) for eliminate the source currents harmonics, reduce the magnitude of neutral wire current (eliminate the zero-sequence current produced by single-phase nonlinear loads), and to compensate the reactive power in the three-phase four-wire electrical networks. This strategy is proposed in order to gate switching pulses generation, dc bus voltage capacitors balancing (conserve equal voltage of the two dc bus capacitors), and to switching frequency reduced and fixed of inverter switches in same times. A Nonlinear Back Stepping Controllers (NBSC) are used for regulated the dc bus voltage capacitors and the SAPF injected currents to robustness, stabilizing the system and to improve the response and to eliminate the overshoot and undershoot of traditional PI (Proportional-Integral). Conventional three-level three dimensional space vector modulation (C3L-3DSVM) and B3L-3DSVM are calculated and compared in terms of error between the two dc bus voltage capacitors, SAPF output voltages and THDv, THDi of source currents, magnitude of source neutral wire current, and the reactive power compensation under unbalanced single phase nonlinear loads. The success, robustness, and the effectiveness of the proposed control strategies are demonstrated through simulation using Sim Power Systems and S-Function of MATLAB/SIMULINK. PMID:27018144

  2. Sub-Domains of Ricin’s B Subunit as Targets of Toxin Neutralizing and Non-Neutralizing Monoclonal Antibodies

    PubMed Central

    Yermakova, Anastasiya; Vance, David J.; Mantis, Nicholas J.

    2012-01-01

    The B subunit (RTB) of ricin toxin is a galactose (Gal)−/N-acetylgalactosamine (GalNac)-specific lectin that mediates attachment, entry, and intracellular trafficking of ricin in host cells. Structurally, RTB consists of two globular domains with identical folding topologies. Domains 1 and 2 are each comprised of three homologous sub-domains (α, β, γ) that likely arose by gene duplication from a primordial carbohydrate recognition domain (CRD), although only sub-domains 1α and 2γ retain functional lectin activity. As part of our ongoing effort to generate a comprehensive B cell epitope map of ricin, we report the characterization of three new RTB-specific monoclonal antibodies (mAbs). All three mAbs, JB4, B/J F9 and C/M A2, were initially identified based on their abilities to neutralize ricin in a Vero cell cytotoxicty assay and to partially (or completely) block ricin attachment to cell surfaces. However, only JB4 proved capable of neutralizing ricin in a macrophage apoptosis assay and in imparting passive immunity to mice in a model of systemic intoxication. Using a combination of techniques, including competitive ELISAs, pepscan analysis, differential reactivity by Western blot, as well as affinity enrichment of phage displayed peptides, we tentatively localized the epitopes recognized by the non-neutralizing mAbs B/J F9 and C/M A2 to sub-domains 2α and 2β, respectively. Furthermore, we propose that the epitope recognized by JB4 is within sub-domain 2γ, adjacent to RTB’s high affinity Gal/GalNAc CRD. These data suggest that recognition of RTB’s sub-domains 1α and 2γ are critical determinants of antibody neutralizing activity and protective immunity to ricin. PMID:22984492

  3. Neutral Beam Ion Confinement in NSTX

    SciTech Connect

    D.S. Darrow; E.D. Fredrickson; S.M. Kaye; S.S. Medley; and A.L. Roquemore

    2001-07-24

    Neutral-beam (NB) heating in the National Spherical Torus Experiment (NSTX) began in September 2000 using up to 5 MW of 80 keV deuterium (D) beams. An initial assessment of beam ion confinement has been made using neutron detectors, a neutral particle analyzer (NPA), and a Faraday cup beam ion loss probe. Preliminary neutron results indicate that confinement may be roughly classical in quiescent discharges, but the probe measurements do not match a classical loss model. MHD activity, especially reconnection events (REs) causes substantial disturbance of the beam ion population.

  4. Activation Likelihood Estimation meta-analysis revisited

    PubMed Central

    Eickhoff, Simon B.; Bzdok, Danilo; Laird, Angela R.; Kurth, Florian; Fox, Peter T.

    2011-01-01

    A widely used technique for coordinate-based meta-analysis of neuroimaging data is activation likelihood estimation (ALE), which determines the convergence of foci reported from different experiments. ALE analysis involves modelling these foci as probability distributions whose width is based on empirical estimates of the spatial uncertainty due to the between-subject and between-template variability of neuroimaging data. ALE results are assessed against a null-distribution of random spatial association between experiments, resulting in random-effects inference. In the present revision of this algorithm, we address two remaining drawbacks of the previous algorithm. First, the assessment of spatial association between experiments was based on a highly time-consuming permutation test, which nevertheless entailed the danger of underestimating the right tail of the null-distribution. In this report, we outline how this previous approach may be replaced by a faster and more precise analytical method. Second, the previously applied correction procedure, i.e. controlling the false discovery rate (FDR), is supplemented by new approaches for correcting the family-wise error rate and the cluster-level significance. The different alternatives for drawing inference on meta-analytic results are evaluated on an exemplary dataset on face perception as well as discussed with respect to their methodological limitations and advantages. In summary, we thus replaced the previous permutation algorithm with a faster and more rigorous analytical solution for the null-distribution and comprehensively address the issue of multiple-comparison corrections. The proposed revision of the ALE-algorithm should provide an improved tool for conducting coordinate-based meta-analyses on functional imaging data. PMID:21963913

  5. Photoproduction of neutral pions off protons

    NASA Astrophysics Data System (ADS)

    Crede, V.; Sparks, N.; Wilson, A.; Anisovich, A. V.; Bacelar, J. C. S.; Bantes, R.; Bartholomy, O.; Bayadilov, D.; Beck, R.; Beloglazov, Y. A.; Castelijns, R.; Dutz, H.; Elsner, D.; Ewald, R.; Frommberger, F.; Funke, Chr.; Gregor, R.; Gridnev, A.; Gutz, E.; Hillert, W.; Hoffmeister, P.; Jaegle, I.; Junkersfeld, J.; Kalinowsky, H.; Kammer, S.; Klein, Frank; Klein, Friedrich; Klempt, E.; Kotulla, M.; Krusche, B.; Löhner, H.; Lopatin, I. V.; Lugert, S.; Menze, D.; Mertens, T.; Messchendorp, J. G.; Metag, V.; Nanova, M.; Nikonov, V. A.; Novinski, D.; Novotny, R.; Ostrick, M.; Pant, L. M.; van Pee, H.; Pfeiffer, M.; Roy, A.; Sarantsev, A. V.; Schadmand, S.; Schmidt, C.; Schmieden, H.; Schoch, B.; Shende, S.; Sokhoyan, V.; Süle, A.; Sumachev, V. V.; Szczepanek, T.; Thoma, U.; Trnka, D.; Varma, R.; Walther, D.; Wendel, Ch.

    2011-11-01

    Photoproduction of neutral pions has been studied with the CBELSA/TAPS detector in the reaction γp→pπ0 for photon energies between 0.85 and 2.50 GeV. The π0 mesons are observed in their dominant neutral decay mode: π0→γγ. For the first time, the differential cross sections cover the very forward region, θc.m.<60∘. A partial-wave analysis of these data within the Bonn-Gatchina framework observes the high-mass resonances G17(2190), D13(2080), and D15(2070).

  6. PDX neutral-beam reionization losses

    SciTech Connect

    Kugel, H.W.; Dylla, H.F.; Eubank, H.P.; Kozub, T.A.; Moore, R.; Schilling, G.; Stewart, L.D.; von Halle, A.; Williams, M.D.

    1982-02-01

    Reionization losses for 1.5 MW H/sup 0/ and 2 MW D/sup 0/ neutral beams injected into the PDX tokamak were studied using pressure gauges, photo-transistors, thermocouples, surface shielding, and surface sample analysis. Considerable outgassing of conventionally prepared 304SS ducts occurred during initial injections and gradually decreased with the cumulative absorption of beam power. Reionization power losses are presently about 5% in the ducts and about 12% total for a beamline including the duct. Present duct pressures are attributed primarily to gas from the ion source and neutralizer with much smaller contributions from residual wall desorption. Physical mechanisms for the observed duct outgassing are discussed.

  7. Ergonomically neutral arm support system

    DOEpatents

    Siminovitch, Michael J; Chung, Jeffrey Y; Dellinges, Steven; Lafever, Robin E

    2005-08-02

    An ergonomic arm support system maintains a neutral position for the forearm. A mechanical support structure attached to a chair or other mounting structure supports the arms of a sitting or standing person. The system includes moving elements and tensioning elements to provide a dynamic balancing force against the forearms. The support structure is not fixed or locked in a rigid position, but is an active dynamic system that is maintained in equipoise by the continuous operation of the opposing forces. The support structure includes an armrest connected to a flexible linkage or articulated or pivoting assembly, which includes a tensioning element such as a spring. The pivoting assembly moves up and down, with the tensioning element providing the upward force that balances the downward force of the arm.

  8. Neutralization and biodegradation of sulfur mustard

    SciTech Connect

    Harvey, S.P.; Beaudry, W.T.; Szafraniec, L.L.

    1995-12-31

    One technology recommended for consideration for the disposal of the U.S. Chemical Stockpile is chemical neutralization followed by biodegradation. In the case of sulfur mustard ({open_quotes}mustard gas{close_quotes}, 2,2{prime}-dichlorodiethyl sulfide), alkaline hydrolysis yields a detoxified and biodegradable product. The hydrolysis reaction was studied with respect to the effects of temperature and sulfur mustard concentration on the rate and products of the reaction. A 28-fold overall rate enhancement was observed at 70{degrees}C vs. 30{degrees}C corresponding to an enthalpy of activation value of 17.9 Kcal/mole. Material balance studies conducted by {sup 1}H Nuclear Magnetic Resonance analysis showed that the products of the reaction consisted of thiodiglycol was relatively greater at lower sulfur mustard concentrations and higher temperatures. As temperatures were decreased or sulfur mustard concentrations was increased, the proportion of ether-type compounds increased accordingly. Conditions of 1% (vol//vol) sulfur mustard, 5% stoichiometric excess of NaOH and 90{degrees}C were selected for generation of the hydrolyzed bioreactor influent material. The bioreactor was seeded with activated sludge and was initially operated as 5 liter sequencing batch reactor with a hydraulic residence time of approximately days. Early results show total organic carbon removal of greater than 90%.

  9. Phenomenology of neutral heavy leptons

    SciTech Connect

    Kalyniak, P.; Melo, I.

    1997-02-01

    We continue our previous work on the flavor-conserving leptonic decays of the Z boson with neutral heavy leptons (NHL`s) in the loops by considering box, vertex, and self-energy diagrams for the muon decay. By inclusion of these loops (they contribute to the input parameter M{sub W}), we can probe the full parameter space spanned by the so-called flavor-conserving mixing parameters ee{sub mix},{mu}{mu}{sub mix},{tau}{tau}{sub mix}. We show that only two diagrams from each class (box, vertex, and self-energy) are important; further, after renormalization only two box diagrams {open_quotes}survive{close_quotes} as dominant. We compare the results of our analysis with the existing work in this field and conclude that flavor-conserving decays have certain advantages over traditionally considered flavor-violating ones. {copyright} {ital 1997} {ital The American Physical Society}

  10. Phenomenology of neutral heavy leptons

    NASA Astrophysics Data System (ADS)

    Kalyniak, Pat; Melo, I.

    1997-02-01

    We continue our previous work on the flavor-conserving leptonic decays of the Z boson with neutral heavy leptons (NHL's) in the loops by considering box, vertex, and self-energy diagrams for the muon decay. By inclusion of these loops (they contribute to the input parameter MW), we can probe the full parameter space spanned by the so-called flavor-conserving mixing parameters eemix,μμmix,ττmix. We show that only two diagrams from each class (box, vertex, and self-energy) are important; further, after renormalization only two box diagrams ``survive'' as dominant. We compare the results of our analysis with the existing work in this field and conclude that flavor-conserving decays have certain advantages over traditionally considered flavor-violating ones.

  11. Initial study of neutral post-instruction responses on the Maryland Physics Expectation Survey

    NASA Astrophysics Data System (ADS)

    Saltzman, J.; Price, M. F.; Rogers, M. B.

    2016-06-01

    Epistemological studies generally focus on how students think about their construction of knowledge compared to how experts think about the same ideas. Instruments such as the MPEX and CLASS use a Likert scale to gauge whether students agree or disagree with how experts think about the same ideas. During analysis, five point scale responses are typically reduced to favorable, neutral, and unfavorable with neutral being treated as a nonresponse. What if students are actively selecting neutral and not treating it as a "does not apply?" To address this question we chose to analyze the postinstruction neutral responses of students in our Physics I course using data from multiple years, multiple sections, and multiple instructors. We found that classroom average postinstruction neutral responses were consistently within a band of 15%-25% and that this was also consistent with other published results. It is not yet clear what this pattern means. Is this a measure of students receiving mixed messages from instructors or a measure of a transitional stage that students go through when learning how to be a good college physics student? These initial findings are interesting enough that we are presenting them here with a more detailed question-by-question analysis to be published in the near future. For example, high levels of neutral responses to applied questions (e.g., "All I need to do is. …") may indicate that students are receiving mixed messages from instructors. On the other hand, high levels of neutral responses to conceptual questions (e.g., "Knowledge in physics…") may indicate that students are in a transitional stage between novice and expert.

  12. HIV-1 Antibody Neutralization Breadth Is Associated with Enhanced HIV-Specific CD4+ T Cell Responses

    PubMed Central

    Soghoian, Damien Z.; Lindqvist, Madelene; Ghebremichael, Musie; Donaghey, Faith; Carrington, Mary; Seaman, Michael S.; Kaufmann, Daniel E.; Walker, Bruce D.

    2015-01-01

    ABSTRACT Antigen-specific CD4+ T helper cell responses have long been recognized to be a critical component of effective vaccine immunity. CD4+ T cells are necessary to generate and maintain humoral immune responses by providing help to antigen-specific B cells for the production of antibodies. In HIV infection, CD4+ T cells are thought to be necessary for the induction of Env-specific broadly neutralizing antibodies. However, few studies have investigated the role of HIV-specific CD4+ T cells in association with HIV neutralizing antibody activity in vaccination or natural infection settings. Here, we conducted a comprehensive analysis of HIV-specific CD4+ T cell responses in a cohort of 34 untreated HIV-infected controllers matched for viral load, with and without neutralizing antibody breadth to a panel of viral strains. Our results show that the breadth and magnitude of Gag-specific CD4+ T cell responses were significantly higher in individuals with neutralizing antibodies than in those without neutralizing antibodies. The breadth of Gag-specific CD4+ T cell responses was positively correlated with the breadth of neutralizing antibody activity. Furthermore, the breadth and magnitude of gp41-specific, but not gp120-specific, CD4+ T cell responses were significantly elevated in individuals with neutralizing antibodies. Together, these data suggest that robust Gag-specific CD4+ T cells and, to a lesser extent, gp41-specific CD4+ T cells may provide important intermolecular help to Env-specific B cells that promote the generation or maintenance of Env-specific neutralizing antibodies. IMPORTANCE One of the earliest discoveries related to CD4+ T cell function was their provision of help to B cells in the development of antibody responses. Yet little is known about the role of CD4+ T helper responses in the setting of HIV infection, and no studies to date have evaluated the impact of HIV-specific CD4+ T cells on the generation of antibodies that can neutralize

  13. The control of powerful neutral beams

    SciTech Connect

    Theil, E.; Jacobson, V.

    1986-05-02

    While significant progress has been made in the development of neutral beams for the heating and sustaining of plasmas in large fusion experiments, the control of such devices has largely been a matter of hardware interlocks and operator experience. The need for computer-assisted control becomes more evident, however, with the initiation of multi-beamline experiments. This paper describes a software system that incorporates simple mathematical models coupled to Kalman filters for control of the high power (6 to 8 MW) beams currently under development at Lawrence Berkeley Laboratory's Neutral Beam Engineering Test Facility. Among the principal features of the system are: reduction of a large number of operator variables to just a few (usually one or two); the ability to describe most of the major neutral beams in use and under development; a foundation resting on statistical data analysis and control system principles rather than rules-of-thumb.

  14. Analysis of mitogen-activated protein kinase activity in yeast.

    PubMed

    Elion, Elaine A; Sahoo, Rupam

    2010-01-01

    Mitogen-activated protein (MAP) kinases play central roles in transmitting extracellular and intracellular information in a wide variety of situations in eukaryotic cells. Their activities are perturbed in a large number of diseases, and their activating kinases are currently therapeutic targets in cancer. MAPKs are highly conserved among all eukaryotes. MAPKs were first cloned from the yeast Saccharomyces cerevisiae. Yeast has five MAPKs and one MAPK-like kinase. The mating MAPK Fus3 is the best characterized yeast MAPK. Members of all subfamilies of human MAPKs can functionally substitute S. cerevisiae MAPKs, providing systems to use genetic approaches to study the functions of either yeast or human MAPKs and to identify functionally relevant amino acid residues that enhance or reduce the effects of therapeutically relevant inhibitors and regulatory proteins. Here, we describe an assay to measure Fus3 activity in immune complexes prepared from S. cerevisiae extracts. The assay conditions are applicable to other MAPKs, as well. PMID:20811996

  15. Antihypertensive neutral lipid

    SciTech Connect

    Snyder, F.L.; Blank, M.L.

    1986-06-17

    A method is described for treating a warm-blooded animal comprising administering to the animal a neutral glycerolipid with a 12 to 20 carbon alkyl group at the sn-1 position, a short carbon chain acyl group at the sn-2 position and a hydroxyl group at the sn-3 position in an amount sufficient to lower the arterial blood pressure of the animal. A method is also described for treating a warm-blooded animal comprising administering a composition consisting essentially of a 1-alkyl-2-acetyl (or propionyl)-sn glycerol in combination with a 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine, wherein the 1-alkyl groups contain 12 to 20 carbon atoms, dissolved in an inert pharmaceutically acceptable solvent in amounts sufficient to lower the arterial blood pressure of the animal.

  16. Neutralizing monoclonal antibodies to an extracellular Pseudomonas cepacia protease.

    PubMed Central

    Kooi, C; Cox, A; Darling, P; Sokol, P A

    1994-01-01

    Pseudomonas cepacia produces at least two extracellular proteases with apparent molecular masses of 36,000 and 40,000 Da. The 36-kDa protease has high proteolytic activity and the 40-kDa protease has low proteolytic activity with hide powder azure as a substrate. Monoclonal antibodies (MAbs) were raised against the purified 36- and 40-kDa proteases. Several MAbs directed against the 36-kDa protease were found to recognize the 40-kDa protease by Western immunoblot analysis. Similarly, a MAb directed against the 40-kDa protease recognized the 36-kDa protease, suggesting that these two proteases may be immunologically related. A MAb directed against the 36-kDa protease, designated 36-6-8, and a MAb directed against the 40-kDa protease (MAb G-11) cross-reacted with other extracellular proteases, such as Pseudomonas aeruginosa elastase and alkaline protease, Pseudomonas pseudomallei protease, and the Vibrio cholerae hemagglutinin/protease. MAb 36-6-8 neutralized the P. cepacia 36-kDa protease, P. aeruginosa elastase, P. pseudomallei protease, and V. cholerae hemagglutinin/protease but did not affect P. aeruginosa alkaline protease activity. In contrast, MAb G-11 to the 40-kDa protease neutralized only the P. cepacia 36-kDa protease. This evidence suggests that the neutralizing MAb, 36-6-8, recognizes an epitope conserved among some metalloproteases. This epitope may lie at or near the active site of the P. cepacia 36-kDa protease and P. aeruginosa elastase. Images PMID:7516312

  17. Transient ion neutralization by electrons.

    NASA Technical Reports Server (NTRS)

    Wilhelm, H. E.

    1973-01-01

    The nonlinear initial-boundary-value problems describing the lateral neutralization of ion beams for the cases that (1) an auxiliary electric field accelerates the electrons into the ion space, and (2) the electrons are injected into the ion space at a prescribed current density are treated. Analytical solutions are derived which give the position and speed of the neutralization front as a function of time, and the temporal development of the electron density, velocity, and electric fields during the neutralization process.

  18. Chemistry of carotenoid neutral radicals.

    PubMed

    Ligia Focsan, A; Magyar, Adam; Kispert, Lowell D

    2015-04-15

    Proton loss from the carotenoid radical cations (Car(+)) to form neutral radicals (#Car) was investigated by numerous electrochemical, EPR, ENDOR and DFT studies described herein. The radical cation and neutral radicals were formed in solution electrochemically and stabilized on solid silica-alumina and MCM-41 matrices. Carotenoid neutral radicals were recently identified in Arabidopsis thaliana plant and photosystem II samples. Deprotonation at the terminal ends of a zeaxanthin radical cation could provide a secondary photoprotection pathway which involves quenching excited state chlorophyll by the long-lived zeaxanthin neutral radicals formed. PMID:25687648

  19. A proposed neutral line signature

    NASA Technical Reports Server (NTRS)

    Doxas, I.; Speiser, T. W.; Dusenbery, P. B.; Horton, W.

    1992-01-01

    An identifying signature is proposed for the existence and location of the neutral line in the magnetotail. The signature, abrupt density, and temperature changes in the Earthtail direction, was first discovered in test particle simulations. Such temperature variations have been observed in ISEE data (Huang et. al. 1992), but their connection to the possible existence of a neutral line in the tail has not yet been established. The proposed signature develops earlier than the ion velocity space ridge of Martin and Speiser (1988), but can only be seen by spacecraft in the vicinity of the neutral line, while the latter can locate a neutral line remotely.

  20. Single Cell Analysis of Transcriptional Activation Dynamics

    PubMed Central

    Rafalska-Metcalf, Ilona U.; Powers, Sara Lawrence; Joo, Lucy M.; LeRoy, Gary; Janicki, Susan M.

    2010-01-01

    Background Gene activation is thought to occur through a series of temporally defined regulatory steps. However, this process has not been completely evaluated in single living mammalian cells. Methodology/Principal Findings To investigate the timing and coordination of gene activation events, we tracked the recruitment of GCN5 (histone acetyltransferase), RNA polymerase II, Brd2 and Brd4 (acetyl-lysine binding proteins), in relation to a VP16-transcriptional activator, to a transcription site that can be visualized in single living cells. All accumulated rapidly with the VP16 activator as did the transcribed RNA. RNA was also detected at significantly more transcription sites in cells expressing the VP16-activator compared to a p53-activator. After α-amanitin pre-treatment, the VP16-activator, GCN5, and Brd2 are still recruited to the transcription site but the chromatin does not decondense. Conclusions/Significance This study demonstrates that a strong activator can rapidly overcome the condensed chromatin structure of an inactive transcription site and supercede the expected requirement for regulatory events to proceed in a temporally defined order. Additionally, activator strength determines the number of cells in which transcription is induced as well as the extent of chromatin decondensation. As chromatin decondensation is significantly reduced after α-amanitin pre-treatment, despite the recruitment of transcriptional activation factors, this provides further evidence that transcription drives large-scale chromatin decondensation. PMID:20422051

  1. Structural analysis of underivatized neutral human milk oligosaccharides in the negative ion mode by nano-electrospray MS(n) (part 1: methodology).

    PubMed

    Pfenninger, Anja; Karas, Michael; Finke, Berndt; Stahl, Bernd

    2002-11-01

    Underivatized neutral oligosaccharides from human milk were analyzed by nano-electrospray ionization (ESI) using a quadrupole ion trap mass spectrometer (QIT-MS) in the negative-ion mode. Under these conditions neutral oligosaccharides are observed as deprotonated molecules [M-H]- with high intensity. CID-experiments of these species with the charge localized at the reducing end lead to C-type fragment ions forming a "new" reducing end. Fragmentations are accompanied by cross-ring cleavages that yield information about linkages of internal monosaccharides. Several isomeric compounds with distinct structural features, such as different glycosidic linkages, fucosylation and branching sites were investigated. The rules governing the fragmentation behavior of this class of oligosaccharides were elucidated and tested for a representative number of certain isomeric glycoforms using the MS/MS and MS(n) capabilities of the QIT. On the basis of the specific fragmentation behavior of deprotonated molecules, the position of fucoses and the linkage type (Gal beta-->3 GlcNAc or Gal beta1-->4 GlcNAc) could be determined and linear and branched could be differentiated. Rules could be established which can be applied in further investigations of these types of oligosaccharides even from heterogenous mixtures. PMID:12443024

  2. Intruder Activity Analysis under Unreliable Sensor Networks

    SciTech Connect

    Tae-Sic Yoo; Humberto E. Garcia

    2007-09-01

    This paper addresses the problem of counting intruder activities within a monitored domain by a sensor network. The deployed sensors are unreliable. We characterize imperfect sensors with misdetection and false-alarm probabilities. We model intruder activities with Markov Chains. A set of Hidden Markov Models (HMM) models the imperfect sensors and intruder activities to be monitored. A novel sequential change detection/isolation algorithm is developed to detect and isolate a change from an HMM representing no intruder activity to another HMM representing some intruder activities. Procedures for estimating the entry time and the trace of intruder activities are developed. A domain monitoring example is given to illustrate the presented concepts and computational procedures.

  3. NEUTRAL-BEAM INJECTION

    SciTech Connect

    Kunkel, W.B.

    1980-06-01

    The emphasis in the preceding chapters has been on magnetic confinement of high temperature plasmas. The question of production and heating of such plasmas has been dealt with relatively more briefly. It should not be inferred, however, that these matters must therefore be either trivial or unimportant. A review of the history reveals that in the early days all these aspects of the controlled fusion problem were considered to be on a par, and were tackled simultaneously and with equal vigor. Only the confinement problem turned out to be much more complex than initially anticipated, and richer in challenge to the plasma physicist than the questions of plasma production and heating. On the other hand, the properties of high-temperature plasmas and plasma confinement can only be studied experimentally after the problems of production and of heating to adequate temperatures are solved. It is the purpose of this and the next chapter to supplement the preceding discussions with more detail on two important subjects: neutral-beam injection and radio-frequency heating. These are the major contenders for heating in present and future tokamak and mirror fusion experiments, and even in several proposed reactors. For neutral beams we emphasize here the technology involved, which has undergone a rather remarkable development. The physics of particle and energy deposition in the plasma, and the discussion of the resulting effects on the confined plasma, have been included in previous chapters, and some experimental results are quoted there. Other heating processes of relevance to fusion are mentioned elsewhere in this book, in connection with the experiments where they are used: i.e. ohmic heating, adiabatic compression heating, and alpha-particle heating in Chapter 3 by H.P. Furth; more ohmic heating in Chapter 7, and shock-implosion heating, laser heating, and relativistic-electron beam heating in Chapter 8, both by W. E. Quinn. These methods are relatively straightforward in

  4. CO2-neutral fuels

    NASA Astrophysics Data System (ADS)

    Goede, A. P. H.

    2015-08-01

    The need for storage of renewable energy (RE) generated by photovoltaic, concentrated solar and wind arises from the fact that supply and demand are ill-matched both geographically and temporarily. This already causes problems of overcapacity and grid congestion in countries where the fraction of RE exceeds the 20% level. A system approach is needed, which focusses not only on the energy source, but includes conversion, storage, transport, distribution, use and, last but not least, the recycling of waste. Furthermore, there is a need for more flexibility in the energy system, rather than relying on electrification, integration with other energy systems, for example the gas network, would yield a system less vulnerable to failure and better adapted to requirements. For example, long-term large-scale storage of electrical energy is limited by capacity, yet needed to cover weekly to seasonal demand. This limitation can be overcome by coupling the electricity net to the gas system, considering the fact that the Dutch gas network alone has a storage capacity of 552 TWh, sufficient to cover the entire EU energy demand for over a month. This lecture explores energy storage in chemicals bonds. The focus is on chemicals other than hydrogen, taking advantage of the higher volumetric energy density of hydrocarbons, in this case methane, which has an approximate 3.5 times higher volumetric energy density. More importantly, it allows the ready use of existing gas infrastructure for energy storage, transport and distribution. Intermittent wind electricity generated is converted into synthetic methane, the Power to Gas (P2G) scheme, by splitting feedstock CO2 and H2O into synthesis gas, a mixture of CO and H2. Syngas plays a central role in the synthesis of a range of hydrocarbon products, including methane, diesel and dimethyl ether. The splitting is accomplished by innovative means; plasmolysis and high-temperature solid oxygen electrolysis. A CO2-neutral fuel cycle is

  5. [Distribution pattern of neutral sugar in forest soils along an altitude gradient in Changbai Mountains, Northeast China].

    PubMed

    Tian, Qiu-Xiang; Zhang, Bin; He, Hong-Bo; Zhang, Xu-Dong; Cheng, Wei-Xin

    2013-07-01

    In July 2010, soil samples were collected from five typical forests (Pinus koraiensis and broadleaved mixed forest, Picea and Abies forest, Larix and Abies forest, Betula ermanii forest, and alpine tundra) along an altitude gradient on the northern slope of Changbai Mountains to investigate the distribution and quantity of neutral sugar in the soils and related affecting factors. The origins of the neutral sugar were differentiated to probe into the biochemical accumulation mechanisms of soil organic matter. There was a significant difference in the neutral sugar content among the forest soils. The relative content of soil neutral sugar' s carbon to soil organic carbon ranged in 80.55-170.63 mg C x g(-1), and tended to be increased with elevated altitude. The multiple regression analysis showed that the mean temperature in growth season was the main factor affecting the relative content of soil neutral sugar, and low temperature was conducive to the accumulation of neutral sugar. The ratio of (galactose + mannose) / (arabinose + xylose) in the five soils was around 1.62-2.28, and had an increasing trend with elevated altitude, illustrating that the contribution of soil microbial neutral sugar to soil organic matter increased with elevated altitude. Soil microbial metabolic quotient declined significantly along elevated altitude, suggesting that in low temperature environment, soil microbial activity decreased but the carbon utilization efficiency enhanced. As a result, a significant portion of decomposed plant residues was transformed into microbial neutral sugar and accumulated stably in soil, and thus, increased the proportion of soil microbial neutral sugar.

  6. High-Throughput Analysis of Enzyme Activities

    SciTech Connect

    Lu, Guoxin

    2007-01-01

    High-throughput screening (HTS) techniques have been applied to many research fields nowadays. Robot microarray printing technique and automation microtiter handling technique allows HTS performing in both heterogeneous and homogeneous formats, with minimal sample required for each assay element. In this dissertation, new HTS techniques for enzyme activity analysis were developed. First, patterns of immobilized enzyme on nylon screen were detected by multiplexed capillary system. The imaging resolution is limited by the outer diameter of the capillaries. In order to get finer images, capillaries with smaller outer diameters can be used to form the imaging probe. Application of capillary electrophoresis allows separation of the product from the substrate in the reaction mixture, so that the product doesn't have to have different optical properties with the substrate. UV absorption detection allows almost universal detection for organic molecules. Thus, no modifications of either the substrate or the product molecules are necessary. This technique has the potential to be used in screening of local distribution variations of specific bio-molecules in a tissue or in screening of multiple immobilized catalysts. Another high-throughput screening technique is developed by directly monitoring the light intensity of the immobilized-catalyst surface using a scientific charge-coupled device (CCD). Briefly, the surface of enzyme microarray is focused onto a scientific CCD using an objective lens. By carefully choosing the detection wavelength, generation of product on an enzyme spot can be seen by the CCD. Analyzing the light intensity change over time on an enzyme spot can give information of reaction rate. The same microarray can be used for many times. Thus, high-throughput kinetic studies of hundreds of catalytic reactions are made possible. At last, we studied the fluorescence emission spectra of ADP and obtained the detection limits for ADP under three different

  7. Signs of neutralization in a redundant gene involved in homologous recombination in Wolbachia endosymbionts.

    PubMed

    Badawi, Myriam; Giraud, Isabelle; Vavre, Fabrice; Grève, Pierre; Cordaux, Richard

    2014-09-17

    Genomic reduction in bacterial endosymbionts occurs through large genomic deletions and long-term accumulation of mutations. The latter process involves successive steps including gene neutralization, pseudogenization, and gradual erosion until complete loss. Although many examples of pseudogenes at various levels of degradation have been reported, neutralization cases are scarce because of the transient nature of the process. Gene neutralization may occur due to relaxation of selection in nonessential genes, for example, those involved in redundant functions. Here, we report an example of gene neutralization in the homologous recombination (HR) pathway of Wolbachia, a bacterial endosymbiont of arthropods and nematodes. The HR pathway is often depleted in endosymbiont genomes, but it is apparently intact in some Wolbachia strains. Analysis of 12 major HR genes showed that they have been globally under strong purifying selection during the evolution of Wolbachia strains hosted by arthropods, supporting the evolutionary importance of the HR pathway for these Wolbachia genomes. However, we detected signs of recent neutralization of the ruvA gene in a subset of Wolbachia strains, which might be related to an ancestral, clade-specific amino acid change that impaired DNA-binding activity. Strikingly, RuvA is part of the RuvAB complex involved in branch migration, whose function overlaps with the RecG helicase. Although ruvA is experiencing neutralization, recG is under strong purifying selection. Thus, our high phylogenetic resolution suggests that we identified a rare example of targeted neutralization of a gene involved in a redundant function in an endosymbiont genome.

  8. Signs of Neutralization in a Redundant Gene Involved in Homologous Recombination in Wolbachia Endosymbionts

    PubMed Central

    Badawi, Myriam; Giraud, Isabelle; Vavre, Fabrice; Grève, Pierre; Cordaux, Richard

    2014-01-01

    Genomic reduction in bacterial endosymbionts occurs through large genomic deletions and long-term accumulation of mutations. The latter process involves successive steps including gene neutralization, pseudogenization, and gradual erosion until complete loss. Although many examples of pseudogenes at various levels of degradation have been reported, neutralization cases are scarce because of the transient nature of the process. Gene neutralization may occur due to relaxation of selection in nonessential genes, for example, those involved in redundant functions. Here, we report an example of gene neutralization in the homologous recombination (HR) pathway of Wolbachia, a bacterial endosymbiont of arthropods and nematodes. The HR pathway is often depleted in endosymbiont genomes, but it is apparently intact in some Wolbachia strains. Analysis of 12 major HR genes showed that they have been globally under strong purifying selection during the evolution of Wolbachia strains hosted by arthropods, supporting the evolutionary importance of the HR pathway for these Wolbachia genomes. However, we detected signs of recent neutralization of the ruvA gene in a subset of Wolbachia strains, which might be related to an ancestral, clade-specific amino acid change that impaired DNA-binding activity. Strikingly, RuvA is part of the RuvAB complex involved in branch migration, whose function overlaps with the RecG helicase. Although ruvA is experiencing neutralization, recG is under strong purifying selection. Thus, our high phylogenetic resolution suggests that we identified a rare example of targeted neutralization of a gene involved in a redundant function in an endosymbiont genome. PMID:25230723

  9. H7N9 influenza virus neutralizing antibodies that possess few somatic mutations

    PubMed Central

    Thornburg, Natalie J.; Zhang, Heng; Bangaru, Sandhya; Kose, Nurgun; Lampley, Rebecca M.; Bombardi, Robin G.; Yu, Yingchun; Graham, Stephen; Branchizio, Andre; Yoder, Sandra M.; Rock, Michael T.; Creech, C. Buddy; Edwards, Kathryn M.; Lee, David; Li, Sheng; Wilson, Ian A.; García-Sastre, Adolfo; Albrecht, Randy A.; Crowe, James E.

    2016-01-01

    Avian H7N9 influenza viruses are group 2 influenza A viruses that have been identified as the etiologic agent for a current major outbreak that began in China in 2013 and may pose a pandemic threat. Here, we examined the human H7-reactive antibody response in 75 recipients of a monovalent inactivated A/Shanghai/02/2013 H7N9 vaccine. After 2 doses of vaccine, the majority of donors had memory B cells that secreted IgGs specific for H7 HA, with dominant responses against single HA subtypes, although frequencies of H7-reactive B cells ranged widely between donors. We isolated 12 naturally occurring mAbs with low half-maximal effective concentrations for binding, 5 of which possessed neutralizing and HA-inhibiting activities. The 5 neutralizing mAbs exhibited narrow breadth of reactivity with influenza H7 strains. Epitope-mapping studies using neutralization escape mutant analysis, deuterium exchange mass spectrometry, and x-ray crystallography revealed that these neutralizing mAbs bind near the receptor-binding pocket on HA. All 5 neutralizing mAbs possessed low numbers of somatic mutations, suggesting the clones arose from naive B cells. The most potent mAb, H7.167, was tested as a prophylactic treatment in a mouse intranasal virus challenge study, and systemic administration of the mAb markedly reduced viral lung titers. PMID:26950424

  10. Fine mapping of sequential neutralization epitopes on the subunit protein VP8 of human rotavirus.

    PubMed Central

    Kovacs-Nolan, Jennifer; Yoo, Dongwan; Mine, Yoshinori

    2003-01-01

    The epitopes of the HRV (human rotavirus), especially those involved in virus neutralization, have not been determined in their entirety, and would have significant implications for HRV vaccine development. In the present study, we report on the epitope mapping and identification of sequential neutralization epitopes, on the Wa strain HRV subunit protein VP8, using synthetic overlapping peptides. Polyclonal antibodies against recombinant Wa VP8 were produced previously in chicken, and purified from egg yolk, which showed neutralizing activity against HRV in vitro. Overlapping VP8 peptide fragments were synthesized and probed with the anti-VP8 antibodies, revealing five sequential epitopes on VP8. Further analysis suggested that three of the five epitopes detected, M1-L10, I55-D66 and L223-P234, were involved in virus neutralization, indicating that sequential epitopes may also be important for the HRV neutralization. The interactions of the antibodies with the five epitopes were characterized by an examination of the critical amino acids involved in antibody binding. Epitopes comprised primarily of hydrophobic amino acid residues, followed by polar and charged residues. The more critical amino acids appeared to be located near the centre of the epitopes, with proline, isoleucine, serine, glutamine and arginine playing an important role in the binding of antibody to the VP8 epitopes. PMID:12901721

  11. A novel reporter system for neutralizing and enhancing antibody assay against dengue virus

    PubMed Central

    2014-01-01

    Background Dengue virus (DENV) still poses a global public health threat, and no vaccine or antiviral therapy is currently available. Antibody plays distinct roles in controlling DENV infections. Neutralizing antibody is protective against DENV infection, whereas sub-neutralizing concentration of antibody can increase DENV infection, termed antibody-dependent enhancement (ADE). Plaque-based assay represents the most widely accepted method measuring neutralizing or enhancing antibodies. Results In this study, a novel reporter virus-based system was developed for measuring neutralization and ADE activity. A stable Renilla luciferase reporter DENV (Luc-DENV) that can produce robust luciferase signals in BHK-21 and K562 cells were used to establish the assay and validated against traditional plaque-based assay. Luciferase value analysis using various known DENV-specific monoclonal antibodies showed good repeatability and a well linear correlation with conventional plaque-based assays. The newly developed assay was finally validated with clinical samples from infected animals and individuals. Conclusions This reporter virus-based assay for neutralizing and enhancing antibody evaluation is rapid, lower cost, and high throughput, and will be helpful for laboratory detection and epidemiological investigation for DENV antibodies. PMID:24548533

  12. Potent dengue virus neutralization by a therapeutic antibody with low monovalent affinity requires bivalent engagement.

    PubMed

    Edeling, Melissa A; Austin, S Kyle; Shrestha, Bimmi; Dowd, Kimberly A; Mukherjee, Swati; Nelson, Christopher A; Johnson, Syd; Mabila, Manu N; Christian, Elizabeth A; Rucker, Joseph; Pierson, Theodore C; Diamond, Michael S; Fremont, Daved H

    2014-04-01

    We recently described our most potently neutralizing monoclonal antibody, E106, which protected against lethal Dengue virus type 1 (DENV-1) infection in mice. To further understand its functional properties, we determined the crystal structure of E106 Fab in complex with domain III (DIII) of DENV-1 envelope (E) protein to 2.45 Å resolution. Analysis of the complex revealed a small antibody-antigen interface with the epitope on DIII composed of nine residues along the lateral ridge and A-strand regions. Despite strong virus neutralizing activity of E106 IgG at picomolar concentrations, E106 Fab exhibited a ∼20,000-fold decrease in virus neutralization and bound isolated DIII, E, or viral particles with only a micromolar monovalent affinity. In comparison, E106 IgG bound DENV-1 virions with nanomolar avidity. The E106 epitope appears readily accessible on virions, as neutralization was largely temperature-independent. Collectively, our data suggest that E106 neutralizes DENV-1 infection through bivalent engagement of adjacent DIII subunits on a single virion. The isolation of anti-flavivirus antibodies that require bivalent binding to inhibit infection efficiently may be a rare event due to the unique icosahedral arrangement of envelope proteins on the virion surface. PMID:24743696

  13. Karyotype Analysis Activity: A Constructivist Learning Design

    ERIC Educational Resources Information Center

    Ahmed, Noveera T.

    2015-01-01

    This classroom activity is based on a constructivist learning design and engages students in physically constructing a karyotype of three mock patients. Students then diagnose the chromosomal aneuploidy based on the karyotype, list the symptoms associated with the disorder, and discuss the implications of the diagnosis. This activity is targeted…

  14. Europa's Neutral Gas Torus

    NASA Astrophysics Data System (ADS)

    Mauk, B. H.; Mitchell, D. G.; McEntire, R. W.; Paranicas, C. P.; Roelof, E. C.; Williams, D. J.; Krimigis, S. M.; Lagg, A.

    2004-05-01

    In-situ energetic ion measurements from the Galileo spacecraft and remote energetic neutral atom (ENA) images from the Cassini spacecraft have been previously interpreted as revealing an unexpectedly massive torus of gas co-orbiting with Jupiter's moon Europa (Lagg et al., 2003; Mauk et al., 2003). Here we report on the results of detailed modeling of the ENA emission process from the Europa regions. Updates to the distribution and composition of the trapped energetic ion populations are included in the models, as are considerations of the partitioning of the gas products into multiple atomic and molecular species. Comparisons between the models and the Cassini observations reveal a torus with a total gas content equal to (0.5 +/- 0.2) E34 atoms plus molecules. This value is higher than, but within a factor of 3 of, an estimate inferred from a prediction of gas densities derived from Voyager plasma measurements and modeling of the interaction between the plasmas and the gases assumed to be emanating from Europa (Schreier et al., 1993). Lagg, A., N. Krupp, J. Woch, and D. J. Williams, Geophys. Res. Lett., 30, DOI 10.1029/2003GL017214, 2003. Mauk, B. H., D. G. Mitchell, S. M. Krimigs, E. C. Roelof, and C. P. Paranicas, Nature, 241, 920, 2003. Schreier, S., A. Eviatar, V. M. Vasyliunas, and J. D. Richardson, J. Geophys. Res., 98, 21231, 1993.

  15. The status of neutral currents

    SciTech Connect

    Zwirner, F.

    1987-11-01

    The situation of particle physics today is quite puzzling. On the one hand, the Standard Model (SM) of strong and electroweak interactions is consistent with all confirmed experimental data but theoretically rather unsatisfactory. On the other hand, none of the many theoretical speculations which try to go beyond the SM has (yet) received the slightest experimental support. The solution to this dilemma can only come from new data: either from the detection of a new particle threshold at high energy colliders, or from the appearance of some small discrepancy in high-precision experiments. A crucial sector for testing the SM and its extensions is that of neutral currents (NC), where an impressive amount of data has been collected in recent years. While waiting for the next generation of experiments, it is certainly useful to take stock of our knowledge, determining the NC parameters as precisely as we can and putting limits on possible deviations from the SM. The present talk contains the results of a recent analysis along these lines: the first part illustrates how a set of 'model-independent' parameters can be extracted from the available NC data, the second part particularizes the analysis to the SM and to some superstring-inspired models with an additional Z' in their low-energy spectrum. 27 refs., 3 figs., 1 tab.

  16. Neutralization tests on the SERT 2 spacecraft

    NASA Technical Reports Server (NTRS)

    Kerslake, W. R.; Domitz, S.

    1979-01-01

    Neutralization test data obtained on the SERT 2 spacecraft are presented. Tests included ion beam neutralization of a thruster by a close (normal design) neutralizer as well as by a distant (1 meter) neutralizer. Parameters affecting neutralization, such as neutralizer bias voltage, neutralizer anode voltage, local spacecraft plasma density, and solar array voltage configuration were varied and changes in plasma potentials were measured. A plasma model is presented as an approximation of observed results.

  17. The IKK-neutralizing compound Bay11 kills supereffector CD8 T cells by altering caspase-dependent activation-induced cell death.

    PubMed

    Lee, Seung-Joo; Long, Meixiao; Adler, Adam J; Mittler, Robert S; Vella, Anthony T

    2009-01-01

    Antigen with dual costimulation through CD137 and CD134 induces powerful CD8 T cell responses. These effector T cells are endowed with an intrinsic survival program resulting in their accumulation in vivo, but the signaling components required for survival are unknown. We tested a cadre of pathway inhibitors and found one preclinical compound, Bay11-7082 (Bay11), which prevented survival. Even the gammac cytokine family members IL-2, -4, -7, and -15 could not block death, nor could pretreatment with IL-7. We found that dual costimulation caused loading of phosphorylated IkappaBalpha (p-IkappaBalpha) and high basal levels of NF-kappaB activity in the effector CD8 T cells. Bay11 trumped both events by reducing the presence of p-IkappaBalpha and ensuing NF-kappaB activity. Not all pathways were impacted to this degree, however, as mitogen-mediated ERK phosphorylation was evident during NF-kappaB inhibition. Nonetheless, Bay11 blocked TCR-stimulated cytokine synthesis by rapidly accentuating activation-induced cell death through elicitation of a caspase-independent pathway. Thus, in effector CD8 T cells, Bay11 forces a dominant caspase-independent death signal that cannot be overcome by an intrinsic survival program nor by survival-inducing cytokines. Therefore, Bay11 may be a useful tool to deliberately kill death-resistant effector T cells for therapeutic benefit.

  18. Electrostatics of cell membrane recognition: structure and activity of neutral and cationic rigid push-pull rods in isoelectric, anionic, and polarized lipid bilayer membranes.

    PubMed

    Sakai, N; Gerard, D; Matile, S

    2001-03-21

    Design, synthesis, and structural and functional studies of rigid-rod ionophores of different axial electrostatic asymmetry are reported. The employed design strategy emphasized presence of (a) a rigid scaffold to minimize the conformational complexity, (b) a unimolecular ion-conducting pathway to minimize the suprastructural complexity and monitor the function, (c) an extended fluorophore to monitor structure, (d) variable axial rod dipole, and (e) variable terminal charges to create axial asymmetry. Studies in isoelectric, anionic, and polarized bilayer membranes confirmed a general increase in activity of uncharged rigid push-pull rods in polarized bilayers. The similarly increased activity of cationic rigid push-pull rods with an electrostatic asymmetry comparable to that of alpha-helical bee toxin melittin (positive charge near negative axial dipole terminus) is shown by fluorescence-depth quenching experiments to originate from the stabilization of transmembrane rod orientation by the membrane potential. The reduced activity of rigid push-pull rods having an electrostatic asymmetry comparable to that in alpha-helical natural antibiotics (a positive charge near the positive axial dipole terminus) is shown by structural studies to originate from rod "ejection" by membrane potentials comparable to that found in mammalian plasma membranes. This structural evidence for cell membrane recognition by asymmetric rods is unprecedented and of possible practical importance with regard to antibiotic resistance.

  19. Type- and Subcomplex-Specific Neutralizing Antibodies against Domain III of Dengue Virus Type 2 Envelope Protein Recognize Adjacent Epitopes▿

    PubMed Central

    Sukupolvi-Petty, Soila; Austin, S. Kyle; Purtha, Whitney E.; Oliphant, Theodore; Nybakken, Grant E.; Schlesinger, Jacob J.; Roehrig, John T.; Gromowski, Gregory D.; Barrett, Alan D.; Fremont, Daved H.; Diamond, Michael S.

    2007-01-01

    Neutralization of flaviviruses in vivo correlates with the development of an antibody response against the viral envelope (E) protein. Previous studies demonstrated that monoclonal antibodies (MAbs) against an epitope on the lateral ridge of domain III (DIII) of the West Nile virus (WNV) E protein strongly protect against infection in animals. Based on X-ray crystallography and sequence analysis, an analogous type-specific neutralizing epitope for individual serotypes of the related flavivirus dengue virus (DENV) was hypothesized. Using yeast surface display of DIII variants, we defined contact residues of a panel of type-specific, subcomplex-specific, and cross-reactive MAbs that recognize DIII of DENV type 2 (DENV-2) and have different neutralizing potentials. Type-specific MAbs with neutralizing activity against DENV-2 localized to a sequence-unique epitope on the lateral ridge of DIII, centered at the FG loop near residues E383 and P384, analogous in position to that observed with WNV-specific strongly neutralizing MAbs. Subcomplex-specific MAbs that bound some but not all DENV serotypes and neutralized DENV-2 infection recognized an adjacent epitope centered on the connecting A strand of DIII at residues K305, K307, and K310. In contrast, several MAbs that had poor neutralizing activity against DENV-2 and cross-reacted with all DENV serotypes and other flaviviruses recognized an epitope with residues in the AB loop of DIII, a conserved region that is predicted to have limited accessibility on the mature virion. Overall, our experiments define adjacent and structurally distinct epitopes on DIII of DENV-2 which elicit type-specific, subcomplex-specific, and cross-reactive antibodies with different neutralizing potentials. PMID:17881453

  20. Type- and subcomplex-specific neutralizing antibodies against domain III of dengue virus type 2 envelope protein recognize adjacent epitopes.

    PubMed

    Sukupolvi-Petty, Soila; Austin, S Kyle; Purtha, Whitney E; Oliphant, Theodore; Nybakken, Grant E; Schlesinger, Jacob J; Roehrig, John T; Gromowski, Gregory D; Barrett, Alan D; Fremont, Daved H; Diamond, Michael S

    2007-12-01

    Neutralization of flaviviruses in vivo correlates with the development of an antibody response against the viral envelope (E) protein. Previous studies demonstrated that monoclonal antibodies (MAbs) against an epitope on the lateral ridge of domain III (DIII) of the West Nile virus (WNV) E protein strongly protect against infection in animals. Based on X-ray crystallography and sequence analysis, an analogous type-specific neutralizing epitope for individual serotypes of the related flavivirus dengue virus (DENV) was hypothesized. Using yeast surface display of DIII variants, we defined contact residues of a panel of type-specific, subcomplex-specific, and cross-reactive MAbs that recognize DIII of DENV type 2 (DENV-2) and have different neutralizing potentials. Type-specific MAbs with neutralizing activity against DENV-2 localized to a sequence-unique epitope on the lateral ridge of DIII, centered at the FG loop near residues E383 and P384, analogous in position to that observed with WNV-specific strongly neutralizing MAbs. Subcomplex-specific MAbs that bound some but not all DENV serotypes and neutralized DENV-2 infection recognized an adjacent epitope centered on the connecting A strand of DIII at residues K305, K307, and K310. In contrast, several MAbs that had poor neutralizing activity against DENV-2 and cross-reacted with all DENV serotypes and other flaviviruses recognized an epitope with residues in the AB loop of DIII, a conserved region that is predicted to have limited accessibility on the mature virion. Overall, our experiments define adjacent and structurally distinct epitopes on DIII of DENV-2 which elicit type-specific, subcomplex-specific, and cross-reactive antibodies with different neutralizing potentials.

  1. Association with emotional information alters subsequent processing of neutral faces

    PubMed Central

    Riggs, Lily; Fujioka, Takako; Chan, Jessica; McQuiggan, Douglas A.; Anderson, Adam K.; Ryan, Jennifer D.

    2014-01-01

    The processing of emotional as compared to neutral information is associated with different patterns in eye movement and neural activity. However, the ‘emotionality’ of a stimulus can be conveyed not only by its physical properties, but also by the information that is presented with it. There is very limited work examining the how emotional information may influence the immediate perceptual processing of otherwise neutral information. We examined how presenting an emotion label for a neutral face may influence subsequent processing by using eye movement monitoring (EMM) and magnetoencephalography (MEG) simultaneously. Participants viewed a series of faces with neutral expressions. Each face was followed by a unique negative or neutral sentence to describe that person, and then the same face was presented in isolation again. Viewing of faces paired with a negative sentence was associated with increased early viewing of the eye region and increased neural activity between 600 and 1200 ms in emotion processing regions such as the cingulate, medial prefrontal cortex, and amygdala, as well as posterior regions such as the precuneus and occipital cortex. Viewing of faces paired with a neutral sentence was associated with increased activity in the parahippocampal gyrus during the same time window. By monitoring behavior and neural activity within the same paradigm, these findings demonstrate that emotional information alters subsequent visual scanning and the neural systems that are presumably invoked to maintain a representation of the neutral information along with its emotional details. PMID:25566024

  2. Global Profiling and Novel Structure Discovery Using Multiple Neutral Loss/Precursor Ion Scanning Combined with Substructure Recognition and Statistical Analysis (MNPSS): Characterization of Terpene-Conjugated Curcuminoids in Curcuma longa as a Case Study.

    PubMed

    Qiao, Xue; Lin, Xiong-hao; Ji, Shuai; Zhang, Zheng-xiang; Bo, Tao; Guo, De-an; Ye, Min

    2016-01-01

    To fully understand the chemical diversity of an herbal medicine is challenging. In this work, we describe a new approach to globally profile and discover novel compounds from an herbal extract using multiple neutral loss/precursor ion scanning combined with substructure recognition and statistical analysis. Turmeric (the rhizomes of Curcuma longa L.) was used as an example. This approach consists of three steps: (i) multiple neutral loss/precursor ion scanning to obtain substructure information; (ii) targeted identification of new compounds by extracted ion current and substructure recognition; and (iii) untargeted identification using total ion current and multivariate statistical analysis to discover novel structures. Using this approach, 846 terpecurcumins (terpene-conjugated curcuminoids) were discovered from turmeric, including a number of potentially novel compounds. Furthermore, two unprecedented compounds (terpecurcumins X and Y) were purified, and their structures were identified by NMR spectroscopy. This study extended the application of mass spectrometry to global profiling of natural products in herbal medicines and could help chemists to rapidly discover novel compounds from a complex matrix.

  3. Genetics Home Reference: neutral lipid storage disease with myopathy

    MedlinePlus

    ... inflammation of the pancreas (pancreatitis), reduced thyroid activity (hypothyroidism), and type 2 diabetes mellitus (the most common ... Neutral lipid storage disease with myopathy MedlinePlus Encyclopedia: Hypothyroidism MedlinePlus Encyclopedia: Type 2 Diabetes These resources from ...

  4. Neutral point detection by satellites. [magnetospheric neutral sheets

    NASA Technical Reports Server (NTRS)

    Schindler, K.; Ness, N. F.

    1974-01-01

    The concept of a neutral point depends on the physical phenomena described. The regions with B less than about 1 gamma detected by Schindler and Ness may be interpreted as neutral regions for the ion-tearing process. The assumption of the presence of a multiple neutral point structure (with temporal variations) is still the most promising interpretation of the Explorer 34 data. Alternatives suggested by Russell lead to difficulties. Nevertheless, the final answer can come only from multiple satellite systems. A 1-day displacement of the day count in the data discussed by Schindler and Ness is corrected.

  5. Neutralization of rainwater acidity at Kanpur, India

    NASA Astrophysics Data System (ADS)

    Shukla, Sheo Prasad; Sharma, Mukesh

    2010-07-01

    Particulate matter (PM) levels show significant seasonal variability and this can influence the neutralization of rainwater acidity. Months were grouped in two periods: monsoon (July to October) and non-monsoon (November to June) for studying the seasonal variability in PM and rainwater composition. To clearly establish the cause effect relationship of acid rain neutralization, a two tier model was proposed involving source apportionment of particulates at two levels: (i) ambient air and (ii) rainwater particulate interaction. For modelling purpose, PM10 (n = 100), soil (n = 4) and rainwater (n = 83) samples were collected at Kanpur, India during 2000-2002. The collected samples were analysed for metals and water soluble ion composition to employ factor analysis for source identification. Knowledge of statistical correlation and chemistry fundamentals were combined to estimate the sources for acid rain neutralization. NH4+ was a dominating ion responsible for neutralizing the acidity of rainwater in monsoon period and Ca2+ was dominating in non-monsoon period. Components of secondary particles (SO42- and NO3-) showed affinity with NH4+, signifying the major role that ammonia can play if present in excess of stoichiometric requirements.

  6. Cross-clade neutralizing antibodies against HIV-1 induced in rabbits by focusing the immune response on a neutralizing epitope

    SciTech Connect

    Zolla-Pazner, Susan; Cohen, Sandra; Pinter, Abraham; Krachmarov, Chavdar; Wrin, Terri; Wang Shixia; Lu Shan

    2009-09-15

    Studies were performed to induce cross-clade neutralizing antibodies (Abs) by testing various combinations of prime and boost constructs that focus the immune response on structurally-conserved epitopes in the V3 loop of HIV-1 gp120. Rabbits were immunized with gp120 DNA containing a V3 loop characterized by the GPGR motif at its tip, and/or with gp120 DNA with a V3 loop carrying the GPGQ motif. Priming was followed by boosts with V3-fusion proteins (V3-FPs) carrying the V3 sequence from a subtype B virus (GPGR motif), and/or with V3 sequences from subtypes A and C (GPGQ motif). The broadest and most consistent neutralizing responses were generated when using a clade C gp120 DNA prime and with the V3{sub B}-FP boost. Immune sera displayed neutralizing activity in three assays against pseudoviruses and primary isolates from subtypes A, AG, B, C, and D. Polyclonal Abs in the immune rabbit sera neutralized viruses that were not neutralized by pools of human anti-V3 monoclonal Abs. Greater than 80% of the neutralizing Abs were specific for V3, showing that the immune response could be focused on a neutralizing epitope and that vaccine-induced anti-V3 Abs have cross-clade neutralizing activity.

  7. Overview of MSFC's Applied Fluid Dynamics Analysis Group Activities

    NASA Technical Reports Server (NTRS)

    Garcia, Roberto; Griffin, Lisa; Williams, Robert

    2002-01-01

    This viewgraph report presents an overview of activities and accomplishments of NASA's Marshall Space Flight Center's Applied Fluid Dynamics Analysis Group. Expertise in this group focuses on high-fidelity fluids design and analysis with application to space shuttle propulsion and next generation launch technologies. Topics covered include: computational fluid dynamics research and goals, turbomachinery research and activities, nozzle research and activities, combustion devices, engine systems, MDA development and CFD process improvements.

  8. Acceleration of reverse analysis method using hyperbolic activation function

    NASA Astrophysics Data System (ADS)

    Pwasong, Augustine; Sathasivam, Saratha

    2015-10-01

    Hyperbolic activation function is examined for its ability to accelerate the performance of doing data mining by using a technique named as Reverse Analysis method. In this paper, we describe how Hopfield network perform better with hyperbolic activation function and able to induce logical rules from large database by using reverse analysis method: given the values of the connections of a network, we can hope to know what logical rules are entrenched in the database. We limit our analysis to Horn clauses.

  9. An Analysis of Solar Global Activity

    NASA Astrophysics Data System (ADS)

    Mouradian, Zadig

    2013-02-01

    This article proposes a unified observational model of solar activity based on sunspot number and the solar global activity in the rotation of the structures, both per 11-year cycle. The rotation rates show a variation of a half-century period and the same period is also associated to the sunspot amplitude variation. The global solar rotation interweaves with the observed global organisation of solar activity. An important role for this assembly is played by the Grand Cycle formed by the merging of five sunspot cycles: a forgotten discovery by R. Wolf. On the basis of these elements, the nature of the Dalton Minimum, the Maunder Minimum, the Gleissberg Cycle, and the Grand Minima are presented.

  10. Method for photon activation positron annihilation analysis

    DOEpatents

    Akers, Douglas W.

    2006-06-06

    A non-destructive testing method comprises providing a specimen having at least one positron emitter therein; determining a threshold energy for activating the positron emitter; and determining whether a half-life of the positron emitter is less than a selected half-life. If the half-life of the positron emitter is greater than or equal to the selected half-life, then activating the positron emitter by bombarding the specimen with photons having energies greater than the threshold energy and detecting gamma rays produced by annihilation of positrons in the specimen. If the half-life of the positron emitter is less then the selected half-life, then alternately activating the positron emitter by bombarding the specimen with photons having energies greater then the threshold energy and detecting gamma rays produced by positron annihilation within the specimen.

  11. Faculty Activity Analysis in the Universidad Tecnica Del Estado Campuses.

    ERIC Educational Resources Information Center

    Karadima, Oscar

    An analysis of academic activities of college faculty at the eight campuses of Chile's Universidad Tecnica del Estado was conducted. Activities were grouped into seven categories: direct teaching, indirect teaching, research, community services, faculty development, academic administration, and other activities. Following the narrative…

  12. Emotionally Neutral Stimuli Are Not Neutral in Schizophrenia: A Mini Review of Functional Neuroimaging Studies

    PubMed Central

    Potvin, Stéphane; Tikàsz, Andràs; Mendrek, Adrianna

    2016-01-01

    Reliable evidence shows that schizophrenia patients tend to experience negative emotions when presented with emotionally neutral stimuli. Similarly, several functional neuroimaging studies show that schizophrenia patients have increased activations in response to neutral material. However, results are heterogeneous. Here, we review the functional neuroimaging studies that have addressed this research question. Based on the 36 functional neuroimaging studies that we retrieved, it seems that the increased brain reactivity to neutral stimuli is fairly common in schizophrenia, but that the regions involved vary considerably, apart from the amygdala. Prefrontal and cingulate sub-regions and the hippocampus may also be involved. By contrasts, results in individuals at risk for psychosis are less consistent. In schizophrenia patients, results are less consistent in the case of studies using non-facial stimuli, explicit processing paradigms, and/or event-related designs. This means that human faces may convey subtle information (e.g., trustworthiness) other than basic emotional expressions. It also means that the aberrant brain reactivity to neutral stimuli is less likely to occur when experimental paradigms are too cognitively demanding as well as in studies lacking statistical power. The main hypothesis proposed to account for this increased brain reactivity to neutral stimuli is the aberrant salience hypothesis of psychosis. Other investigators propose that the aberrant brain reactivity to neutral stimuli in schizophrenia results from abnormal associative learning, untrustworthiness judgments, priming effects, and/or reduced habituation to neutral stimuli. In the future, the effects of antipsychotics on this aberrant brain reactivity will need to be determined, as well as the potential implication of sex/gender. PMID:27445871

  13. Emotionally Neutral Stimuli Are Not Neutral in Schizophrenia: A Mini Review of Functional Neuroimaging Studies.

    PubMed

    Potvin, Stéphane; Tikàsz, Andràs; Mendrek, Adrianna

    2016-01-01

    Reliable evidence shows that schizophrenia patients tend to experience negative emotions when presented with emotionally neutral stimuli. Similarly, several functional neuroimaging studies show that schizophrenia patients have increased activations in response to neutral material. However, results are heterogeneous. Here, we review the functional neuroimaging studies that have addressed this research question. Based on the 36 functional neuroimaging studies that we retrieved, it seems that the increased brain reactivity to neutral stimuli is fairly common in schizophrenia, but that the regions involved vary considerably, apart from the amygdala. Prefrontal and cingulate sub-regions and the hippocampus may also be involved. By contrasts, results in individuals at risk for psychosis are less consistent. In schizophrenia patients, results are less consistent in the case of studies using non-facial stimuli, explicit processing paradigms, and/or event-related designs. This means that human faces may convey subtle information (e.g., trustworthiness) other than basic emotional expressions. It also means that the aberrant brain reactivity to neutral stimuli is less likely to occur when experimental paradigms are too cognitively demanding as well as in studies lacking statistical power. The main hypothesis proposed to account for this increased brain reactivity to neutral stimuli is the aberrant salience hypothesis of psychosis. Other investigators propose that the aberrant brain reactivity to neutral stimuli in schizophrenia results from abnormal associative learning, untrustworthiness judgments, priming effects, and/or reduced habituation to neutral stimuli. In the future, the effects of antipsychotics on this aberrant brain reactivity will need to be determined, as well as the potential implication of sex/gender. PMID:27445871

  14. Environmental neutralization of polonium-218

    SciTech Connect

    Goldstein, S.D.; Hopke, P.K.

    1985-01-01

    Previous work has indicated that two mechanisms of neutralization of the singly charged polonium ion exist. Charged Polonium-218 can be neutralized by reacting with oxygen to form a polonium oxide ion with a higher ionization potential than that of the polonium metal and then accepting an electron transferred from a lower ionization potential gas. In this present work, this mechanism has been verified by determining that the polonium oxide has an ionization potential in the range 10.35-10.53 eV. It was also previously reported that /sup 218/Po can be neutralized, in the absence of oxygen, by the scavenging of electrons by a trace gas such as water or nitrogen dioxide and their diffusion to the polonium ion. To verify this second neutralization mechanism, concentrations of nitrogen dioxide in nitrogen in the range of 50 ppb-1 ppm were examined for their ability to neutralize the polonium ion. Complete neutralization of /sup 218/Po was observed at nitrogen dioxide concentrations greater than 700 ppb. For concentrations below 700 ppb, the degree of neutralization was found to increase smoothly with the nitrogen dioxide concentration.

  15. Microscopic Analysis of Activated Sludge. Training Manual.

    ERIC Educational Resources Information Center

    Office of Water Program Operations (EPA), Cincinnati, OH. National Training and Operational Technology Center.

    This training manual presents material on the use of a compound microscope to analyze microscope communities, present in wastewater treatment processes, for operational control. Course topics include: sampling techniques, sample handling, laboratory analysis, identification of organisms, data interpretation, and use of the compound microscope.…

  16. Positional nystagmus showing neutral points.

    PubMed

    Hiruma, Kiyoshi; Numata, Tsutomu

    2004-01-01

    We encountered patients who had their static direction-changing positional nystagmus canceled at about 20-30 degrees yaw head rotation from the supine position. This nystagmus was also canceled when the head was rotated 180 degrees from this position. We call these head positions neutral points. At the neutral points, the cupula of the horizontal semicircular canal of the affected ear is positioned vertical to the gravitational plane and no deflection of the cupula occurs. The positional nystagmus observed (except the neutral points) was thought to occur due to a "heavy cupula" or "light cupula", which may be determined by the specific gravity of its endolymph.

  17. [Neutral Medical Claim Management Committee].

    PubMed

    Komatsu, Mitsuru

    2013-03-01

    The Ibaraki Medical Association established the Committee for Alternative Dispute Resolution called the Neutral Medical Claim Management Committee in 2006. Among 64 claims presented to the committee, 29 were settled through mediation or consultation. Patients were generally satisfied that their claims were considered fairly by the committee and that they were able to talk directly with healthcare professionals. However, some did not consider the committee to be completely neutral. The healthcare professionals involved rated the committee highly because they felt that the processes were neutral and no emotional aspects were involved. PMID:23617190

  18. Viper and cobra venom neutralization by beta-sitosterol and stigmasterol isolated from the root extract of Pluchea indica Less. (Asteraceae).

    PubMed

    Gomes, A; Saha, Archita; Chatterjee, Ipshita; Chakravarty, A K

    2007-09-01

    We reported previously that the methanolic root extract of the Indian medicinal plant Pluchea indica Less. (Asteraceae) could neutralize viper venom-induced action [Alam, M.I., Auddy, B., Gomes, A., 1996. Viper venom neutralization by Indian medicinal plant (Hemidesmus indicus and P. indica) root extracts. Phytother. Res. 10, 58-61]. The present study reports the neutralization of viper and cobra venom by beta-sitosterol and stigmasterol isolated from the root extract of P. indica Less. (Asteraceae). The active fraction (containing the major compound beta-sitosterol and the minor compound stigmasterol) was isolated and purified by silica gel column chromatography and the structure was determined using spectroscopic analysis (EIMS, (1)H NMR, (13)C NMR). Anti-snake venom activity was studied in experimental animals. The active fraction was found to significantly neutralize viper venom-induced lethal, hemorrhagic, defibrinogenation, edema and PLA(2) activity. Cobra venom-induced lethality, cardiotoxicity, neurotoxicity, respiratory changes and PLA(2) activity were also antagonized by the active component. It potentiated commercial snake venom antiserum action against venom-induced lethality in male albino mice. The active fraction could antagonize venom-induced changes in lipid peroxidation and superoxide dismutase activity. This study suggests that beta-sitosterol and stigmasterol may play an important role, along with antiserum, in neutralizing snake venom-induced actions.

  19. A Meta-Analysis of 25 Years of Mood-Creativity Research: Hedonic Tone, Activation, or Regulatory Focus?

    ERIC Educational Resources Information Center

    Baas, Matthijs; De Dreu, Carsten K. W.; Nijstad, Bernard A.

    2008-01-01

    This meta-analysis synthesized 102 effect sizes reflecting the relation between specific moods and creativity. Effect sizes overall revealed that positive moods produce more creativity than mood-neutral controls (r = 0.15), but no significant differences between negative moods and mood-neutral controls (r = -0.03) or between positive and negative…

  20. Communication: Classical threshold law for ion-neutral-neutral three-body recombination

    SciTech Connect

    Pérez-Ríos, Jesús; Greene, Chris H.

    2015-07-28

    A very recently method for classical trajectory calculations for three-body collision [Pérez-Ríos et al., J. Chem. Phys. 140, 044307 (2014)] has been applied to describe ion-neutral-neutral ternary processes for low energy collisions: 0.1 mK–10 mK. As a result, a threshold law for the three-body recombination cross section is obtained and corroborated numerically. The derived threshold law predicts the formation of weakly bound dimers, with binding energies comparable to the collision energy of the collisional partners. In this low energy range, this analysis predicts that molecular ions should dominate over molecular neutrals as the most products formed.

  1. Neutralizing antibodies in Borna disease virus-infected rats.

    PubMed Central

    Hatalski, C G; Kliche, S; Stitz, L; Lipkin, W I

    1995-01-01

    Borna disease is a neurologic syndrome caused by infection with a nonsegmented, negative-strand RNA virus, Borna disease virus. Infected animals have antibodies to two soluble viral proteins, p40 and p23, and a membrane-associated viral glycoprotein, gp18. We examined the time course for the development of neutralization activity and the expression of antibodies to individual viral proteins in sera of infected rats. The appearance of neutralizing activity correlated with the development of immunoreactivity to gp18, but not p40 or p23. Monospecific and monoclonal antibodies to native gp18 and recombinant nonglycosylated gp18 were also found to have neutralizing activity and to immunoprecipitate viral particles or subparticles. These findings suggest that gp18 is likely to be present on the surface of the viral particles and is likely to contain epitopes important for virus neutralization. PMID:7815538

  2. Mass spectrometric analysis of neutral and anionic N-glycans from a Dictyostelium discoideum model for human congenital disorder of glycosylation CDG IL.

    PubMed

    Hykollari, Alba; Balog, Crina I A; Rendić, Dubravko; Braulke, Thomas; Wilson, Iain B H; Paschinger, Katharina

    2013-03-01

    The HL241 mutant strain of the cellular slime mold Dictyostelium discoideum is a potential model for human congenital disorder of glycosylation type IL (ALG9-CDG) and has been previously predicted to possess a lower degree of modification of its N-glycans with anionic moieties than the parental wild-type. In this study, we first showed that this strain has a premature stop codon in its alg9 mannosyltransferase gene compatible with the occurrence of truncated N-glycans. These were subject to an optimized analytical workflow, considering that the mass spectrometry of acidic glycans often presents challenges due to neutral loss and suppression effects. Therefore, the protein-bound N-glycans were first fractionated, after serial enzymatic release, by solid phase extraction. Then primarily single glycan species were isolated by mixed hydrophilic-interaction/anion-exchange or reversed-phase HPLC and analyzed using chemical and enzymatic treatments and MS/MS. We show that protein-linked N-glycans of the mutant are of reduced size as compared to those of wild-type AX3, but still contain core α1,3-fucose, intersecting N-acetylglucosamine, bisecting N-acetylglucosamine, methylphosphate, phosphate, and sulfate residues. We observe that a single N-glycan can carry up to four of these six possible modifications. Due to the improved analytical procedures, we reveal fuller details regarding the N-glycomic potential of this fascinating model organism. PMID:23320427

  3. A shell-neutral modeling approach yields sustainable oyster harvest estimates: a retrospective analysis of the Louisiana state primary seed grounds

    USGS Publications Warehouse

    Soniat, Thomas M.; Klinck, John M.; Powell, Eric N.; Cooper, Nathan; Abdelguerfi, Mahdi; Hofmann, Eileen E.; Dahal, Janak; Tu, Shengru; Finigan, John; Eberline, Benjamin S.; La Peyre, Jerome F.; LaPeyre, Megan K.; Qaddoura, Fareed

    2012-01-01

    A numerical model is presented that defines a sustainability criterion as no net loss of shell, and calculates a sustainable harvest of seed (<75 mm) and sack or market oysters (≥75 mm). Stock assessments of the Primary State Seed Grounds conducted east of the Mississippi from 2009 to 2011 show a general trend toward decreasing abundance of sack and seed oysters. Retrospective simulations provide estimates of annual sustainable harvests. Comparisons of simulated sustainable harvests with actual harvests show a trend toward unsustainable harvests toward the end of the time series. Stock assessments combined with shell-neutral models can be used to estimate sustainable harvest and manage cultch through shell planting when actual harvest exceeds sustainable harvest. For exclusive restoration efforts (no fishing allowed), the model provides a metric for restoration success-namely, shell accretion. Oyster fisheries that remove shell versus reef restorations that promote shell accretion, although divergent in their goals, are convergent in their management; both require vigilant attention to shell budgets.

  4. Linking Temporal Changes in Bacterial Community Structures with the Detection and Phylogenetic Analysis of Neutral Metalloprotease Genes in the Sediments of a Hypereutrophic Lake

    PubMed Central

    Tsuboi, Shun; Yamamura, Shigeki; Imai, Akio; Satou, Takayuki; Iwasaki, Kazuhiro

    2014-01-01

    We investigated spatial and temporal variations in bacterial community structures as well as the presence of three functional proteolytic enzyme genes in the sediments of a hypereutrophic freshwater lake in order to acquire an insight into dynamic links between bacterial community structures and proteolytic functions. Bacterial communities determined from 16S rRNA gene clone libraries markedly changed bimonthly, rather than vertically in the sediment cores. The phylum Firmicutes dominated in the 4–6 cm deep sediment layer sample after August in 2007, and this correlated with increases in interstitial ammonium concentrations (p < 0.01). The Firmicutes clones were mostly composed of the genus Bacillus. npr genes encoding neutral metalloprotease, an extracellular protease gene, were detected after the phylum Firmicutes became dominant. The deduced Npr protein sequences from the retrieved npr genes also showed that most of the Npr sequences used in this study were closely related to those of the genus Bacillus, with similarities ranging from 61% to 100%. Synchronous temporal occurrences of the 16S rRNA gene and Npr sequences, both from the genus Bacillus, were positively associated with increases in interstitial ammonium concentrations, which may imply that proteolysis by Npr from the genus Bacillus may contribute to the marked increases observed in ammonium concentrations in the sediments. Our results suggest that sedimentary bacteria may play an important role in the biogeochemical nitrogen cycle of freshwater lakes. PMID:25130992

  5. The Relationship between Neutralization Techniques and Induced Abortion

    PubMed Central

    Kalateh Sadati, Ahmad; Tabei, Seyed Ziaaddin; Salehzadeh, Hamzeh; Rahnavard, Farnaz; Namavar Jahromi, Bahia; Hemmati, Soroor

    2014-01-01

    Background: Induced abortion is not only a serious threat for women’s health, but also a controversial topic for its ethical and moral problems. We aimed to evaluate the relationship between neutralization techniques and attempting to commit abortion in married women with unintended pregnancy. Methods: After in-depth interviews with some women who had attempted abortion, neutralization themes were gathered. Next, to analyze the data quantitatively, a questionnaire was created including demographic and psychosocial variables specifically related to neutralization. The participants were divided into two groups (abortion and control) of unintended pregnancy and were then compared. Results: Analysis of psychosocial variables revealed a significant difference in the two groups at neutralization, showing that neutralization in the control group (56.97±10.24) was higher than that in the abortion group (44.19±12.44). To evaluate the findings more accurately, we examined the causal factors behind the behaviors of the abortion group. Binary logistic regression showed that among psychosocial factors, neutralization significantly affected abortion (95% CI=1.07-1.35). Conclusion: Despite the network of many factors affecting induced abortion, neutralization plays an important role in reinforcing the tendency to attempt abortion. Furthermore, the decline of religious beliefs, as a result of the secular context of the modern world, seems to have an important role in neutralizing induced abortion. PMID:25349851

  6. Initiation of immune tolerance–controlled HIV gp41 neutralizing B cell lineages

    PubMed Central

    Zhang, Ruijun; Verkoczy, Laurent; Wiehe, Kevin; Alam, S. Munir; Nicely, Nathan I.; Santra, Sampa; Bradley, Todd; Pemble, Charles W.; Zhang, Jinsong; Gao, Feng; Montefiori, David C.; Bouton-Verville, Hilary; Kelsoe, Garnett; Larimore, Kevin; Greenberg, Phillip D.; Parks, Robert; Foulger, Andrew; Peel, Jessica N.; Luo, Kan; Lu, Xiaozhi; Trama, Ashley M.; Vandergrift, Nathan; Tomaras, Georgia D.; Kepler, Thomas B.; Moody, M. Anthony; Liao, Hua-Xin; Haynes, Barton F.

    2016-01-01

    Development of an HIV vaccine is a global priority. A major roadblock to a vaccine is an inability to induce protective broadly neutralizing antibodies (bnAbs). HIV gp41 bnAbs have characteristics that predispose them to be controlled by tolerance. We used gp41 2F5 bnAb germline knock-in mice and macaques vaccinated with immunogens reactive with germline precursors to activate neutralizing antibodies. In germline knock-in mice, bnAb precursors were deleted, with remaining anergic B cells capable of being activated by germline-binding immunogens to make gp41-reactive immunoglobulin M (IgM). Immunized macaques made B cell clonal lineages targeted to the 2F5 bnAb epitope, but 2F5-like antibodies were either deleted or did not attain sufficient affinity for gp41-lipid complexes to achieve the neutralization potency of 2F5. Structural analysis of members of a vaccine-induced antibody lineage revealed that heavy chain complementarity-determining region 3 (HCDR3) hydrophobicity was important for neutralization. Thus, gp41 bnAbs are controlled by immune tolerance, requiring vaccination strategies to transiently circumvent tolerance controls. PMID:27122615

  7. Incomplete Neutralization and Deviation from Sigmoidal Neutralization Curves for HIV Broadly Neutralizing Monoclonal Antibodies.

    PubMed

    McCoy, Laura E; Falkowska, Emilia; Doores, Katie J; Le, Khoa; Sok, Devin; van Gils, Marit J; Euler, Zelda; Burger, Judith A; Seaman, Michael S; Sanders, Rogier W; Schuitemaker, Hanneke; Poignard, Pascal; Wrin, Terri; Burton, Dennis R

    2015-08-01

    The broadly neutralizing HIV monoclonal antibodies (bnMAbs) PG9, PG16, PGT151, and PGT152 have been shown earlier to occasionally display an unusual virus neutralization profile with a non-sigmoidal slope and a plateau at <100% neutralization. In the current study, we were interested in determining the extent of non-sigmoidal slopes and plateaus at <100% for HIV bnMAbs more generally. Using both a 278 panel of pseudoviruses in a CD4 T-cell (U87.CCR5.CXCR4) assay and a panel of 117 viruses in the TZM-bl assay, we found that bnMAbs targeting many neutralizing epitopes of the spike had neutralization profiles for at least one virus that plateaued at <90%. Across both panels the bnMAbs targeting the V2 apex of Env and gp41 were most likely to show neutralization curves that plateaued <100%. Conversely, bnMAbs targeting the high-mannose patch epitopes were less likely to show such behavior. Two CD4 binding site (CD4bs) Abs also showed this behavior relatively infrequently. The phenomenon of incomplete neutralization was also observed in a large peripheral blood mononuclear cells (PBMC)-grown molecular virus clone panel derived from patient viral swarms. In addition, five bnMAbs were compared against an 18-virus panel of molecular clones produced in 293T cells and PBMCs and assayed in TZM-bl cells. Examples of plateaus <90% were seen with both types of virus production with no consistent patterns observed. In conclusion, incomplete neutralization and non-sigmoidal neutralization curves are possible for all HIV bnMAbs against a wide range of viruses produced and assayed in both cell lines and primary cells with implications for the use of antibodies in therapy and as tools for vaccine design. PMID:26267277

  8. Incomplete Neutralization and Deviation from Sigmoidal Neutralization Curves for HIV Broadly Neutralizing Monoclonal Antibodies.

    PubMed

    McCoy, Laura E; Falkowska, Emilia; Doores, Katie J; Le, Khoa; Sok, Devin; van Gils, Marit J; Euler, Zelda; Burger, Judith A; Seaman, Michael S; Sanders, Rogier W; Schuitemaker, Hanneke; Poignard, Pascal; Wrin, Terri; Burton, Dennis R

    2015-08-01

    The broadly neutralizing HIV monoclonal antibodies (bnMAbs) PG9, PG16, PGT151, and PGT152 have been shown earlier to occasionally display an unusual virus neutralization profile with a non-sigmoidal slope and a plateau at <100% neutralization. In the current study, we were interested in determining the extent of non-sigmoidal slopes and plateaus at <100% for HIV bnMAbs more generally. Using both a 278 panel of pseudoviruses in a CD4 T-cell (U87.CCR5.CXCR4) assay and a panel of 117 viruses in the TZM-bl assay, we found that bnMAbs targeting many neutralizing epitopes of the spike had neutralization profiles for at least one virus that plateaued at <90%. Across both panels the bnMAbs targeting the V2 apex of Env and gp41 were most likely to show neutralization curves that plateaued <100%. Conversely, bnMAbs targeting the high-mannose patch epitopes were less likely to show such behavior. Two CD4 binding site (CD4bs) Abs also showed this behavior relatively infrequently. The phenomenon of incomplete neutralization was also observed in a large peripheral blood mononuclear cells (PBMC)-grown molecular virus clone panel derived from patient viral swarms. In addition, five bnMAbs were compared against an 18-virus panel of molecular clones produced in 293T cells and PBMCs and assayed in TZM-bl cells. Examples of plateaus <90% were seen with both types of virus production with no consistent patterns observed. In conclusion, incomplete neutralization and non-sigmoidal neutralization curves are possible for all HIV bnMAbs against a wide range of viruses produced and assayed in both cell lines and primary cells with implications for the use of antibodies in therapy and as tools for vaccine design.

  9. Plasma-Neutral Coupling on the Dark and Bright Sides of Antarctica

    NASA Astrophysics Data System (ADS)

    Chu, X.; Yu, Z.; Fong, W.; Chen, C.; Zhao, J.; Huang, W.; Roberts, B. R.; Fuller-Rowell, T. J.; Richmond, A. D.; Gerrard, A. J.; Weatherwax, A. T.; Gardner, C. S.

    2014-12-01

    The polar mesosphere and thermosphere provide a unique natural laboratory for studying the complex physical, chemical, neutral dynamical and electrodynamics processes in the Earth's atmosphere and space environment. McMurdo (geographic 77.83S, geomagnetic 80S) is located by the poleward edge of the aurora oval; so energetic particles may penetrate into the lower thermosphere and mesosphere along nearly vertical geomagnetic field lines. Lidar observations at McMurdo from December 2010 to 2014 have discovered several neutral atmosphere phenomena closely related to ionosphereic parameters and geomagnetic activity. For example, the diurnal tidal amplitude of temperatures not only increases super-exponentially from 100 to 110 km but also its growth rate becomes larger at larger Kp index. The lidar discovery of neutral iron (Fe) layers with gravity wave signatures in the thermosphere enabled the direct measurements of neutral temperatures from 30 to 170 km, revealing the neutral-ion coupling and aurora-enhanced Joule heating. A lidar 'marathon' of 174-hour continuous observations showed dramatic changes of composition (Fe atoms and ice particles) densities (over 40 times) in the mesopause region and their correlations to solar events. In this paper we will study the plasma-neutral coupling on the dark side of Antarctica via observation analysis and numerical modeling of the thermospheric Fe layers in the 100-200 km. A newly developed thermospheric Fe/Fe+ model is used to quantify how Fe+ ions are transported from their main deposition region to the E-F region and then neutralized to form Fe layers under dark polar conditions. We will also study the plasma-neutral coupling on the bright side of Antarctica via analyzing Fe events in summer. Complementary observations will be combined to show how the extreme changes of Fe layers are related to aurora particle precipitation and visible/sub-visible ice particles. These observations and studies will open new areas of

  10. Identification of a Neutralizing Epitope within Antigenic Domain 5 of Glycoprotein B of Human Cytomegalovirus

    PubMed Central

    Wiegers, Anna-Katharina; Sticht, Heinrich; Winkler, Thomas H.; Britt, William J.

    2014-01-01

    ABSTRACT Human cytomegalovirus (HCMV) is an important, ubiquitous pathogen that causes severe clinical disease in immunocompromised individuals, such as organ transplant recipients and infants infected in utero. The envelope glycoprotein B (gB) of HCMV is a major antigen for the induction of virus-neutralizing antibodies. We have begun to define target structures within gB that are recognized by virus-neutralizing antibodies. Antigenic domain 5 (AD-5) of gB has been identified as an important target for neutralizing antibodies in studies using human monoclonal antibodies (MAbs). Anti-AD-5 MAbs share a target site on gB, despite originating from different, healthy, HCMV-infected donors. Mutational analysis of AD-5 identified tyrosine 280 in combination with other surface-exposed residues (the YNND epitope) as critical for antibody binding. The YNND epitope is strictly conserved among different HCMV strains. Recombinant viruses carrying YNND mutations in AD-5 were resistant to virus-neutralizing MAbs. Competition enzyme-linked immunosorbent assays (ELISAs) with human HCMV-convalescent-phase sera from unselected donors confirmed the conserved antibody response for the YNND epitope in HCMV-infected individuals and, because a significant fraction of the gB AD-5 response was directed against the YNND epitope, further argued that this epitope is a major target of anti-AD-5 antibody responses. In addition, affinity-purified polyclonal anti-AD-5 antibodies prepared from individual sera showed reactivity to AD-5 and neutralization activity toward gB mutant viruses that were similar to those of AD-5-specific MAbs. Taken together, our data indicate that the YNND epitope represents an important target for anti-gB antibody responses as well as for anti-AD-5 virus-neutralizing antibodies. IMPORTANCE HCMV is a major global health concern, and a vaccine to prevent HCMV disease is a widely recognized medical need. Glycoprotein B of HCMV is an important target for neutralizing

  11. Active vision in satellite scene analysis

    NASA Technical Reports Server (NTRS)

    Naillon, Martine

    1994-01-01

    In earth observation or planetary exploration it is necessary to have more and, more autonomous systems, able to adapt to unpredictable situations. This imposes the use, in artificial systems, of new concepts in cognition, based on the fact that perception should not be separated from recognition and decision making levels. This means that low level signal processing (perception level) should interact with symbolic and high level processing (decision level). This paper is going to describe the new concept of active vision, implemented in Distributed Artificial Intelligence by Dassault Aviation following a 'structuralist' principle. An application to spatial image interpretation is given, oriented toward flexible robotics.

  12. Passive versus active mitigation cost analysis

    SciTech Connect

    Parazin, R.J.; Galbraith, J.D.

    1995-04-01

    The scope of this task is to assess the impact of mitigation alternatives for Tanks 241-SY-101 and 241-SY-103 on the Project W-236A Multi-Function Waste Tank Facility. This assessment and other related tasks are part of an Action Plan Path Forward prepared by the Tank Waste Remediation System (TWRS) Life Extension and Transition Program. Task 3.7 of the Action Plan for Project W-236A MWTF analyzed the comparative cost/risk of two hydrogen gas mitigation alternatives (active versus passive) to recommend the most appropriate course of action to resolve the hydrogen gas safety issue. The qualitative success of active mitigation has been demonstrated through Tank 241-SY-101 testing. Passive mitigation has not been demonstrated but will be validated by laboratory test work performed under Task 3.1 of the Action Plan. It is assumed for this assessment that the uncertainties associated with the performance of either alternative is comparable. Determining alternative specific performance measures beyond those noted are not in the scope of this effort.

  13. Stability of Non-Neutral Plasma Cylinder Consisting of Magnetized Cold Electrons and of Small Density Fraction of Ions Born at Rest: Non-Local Analysis

    SciTech Connect

    Yeliseyev, Y. N.

    2009-03-30

    The non-local stability problem of the plasma cylinder, filled with 'cold' magnetized rigidly rotating electrons, and a small density fraction of ions, is solved. The ions are supposed to be born at rest by ionization of background gas. The study is based on the kinetic description of ions. The equilibrium distribution function, taking into account the peculiarity of ions birth, is used. The radial electric field is caused by space charge of non-neutral plasma. The dispersion equation for plasma eigen frequencies is obtained analytically. It is valid within the total admissible range of values of electric and magnetic fields. Normalized eigen frequencies {omega}'/{omega}{sub i} are calculated for the basic azimuth mode m = 1({omega}' {omega}-m{omega}{sub i}{sup +}, {omega}{sub +} = (-{omega}{sub ci}+{omega}{sub i})/2, {omega}{sub i} ({omega}{sub ci}{sup 2}-4eE{sub r}/m{sub i}r){sup 1/2} is called the 'modified' ion cyclotron (MIC) frequency), for the density fraction of ions of atomic nitrogen f N{sub i}/n{sub e} = 0,01 and are presented in graphic form versus parameter 2{omega}{sub pe}{sup 2}/{omega}{sub ce}{sup 2}. The spectra of oscillations {omega}'/{omega}{sub i} consist of the family of electron Trivel-piece--Gould (TG) modes and of the families of MIC modes. The frequencies of MIC modes are located in a small vicinity of harmonics of the MIC frequency {omega}{sub i} above and below the harmonic. The TG modes in non-neutral plasma fall in the region of MIC frequencies {omega}{sub i} and interact strongly with MIC modes. The slow TG modes become unstable near the crossings with non-negative harmonics of MIC frequencies. The instabilities have a resonant character. The lowest radial TG mode has a maximum growth rate at crossing with a zero harmonic of {omega}{sub i} ((Im {omega}'/{omega}{sub i}){sub max}{approx_equal}0,074). The growth rates of MIC modes are much lower ((Im {omega}'/{omega}{sub i}){sub max} < or approx. 0,002). Their instability has a threshold

  14. Stability of Non-Neutral Plasma Cylinder Consisting of Magnetized Cold Electrons and of Small Density Fraction of Ions Born at Rest: Non-Local Analysis

    NASA Astrophysics Data System (ADS)

    Yeliseyev, Y. N.

    2009-03-01

    The non-local stability problem of the plasma cylinder, filled with "cold" magnetized rigidly rotating electrons, and a small density fraction of ions, is solved. The ions are supposed to be born at rest by ionization of background gas. The study is based on the kinetic description of ions. The equilibrium distribution function, taking into account the peculiarity of ions birth, is used. The radial electric field is caused by space charge of non-neutral plasma. The dispersion equation for plasma eigen frequencies is obtained analytically. It is valid within the total admissible range of values of electric and magnetic fields. Normalized eigen frequencies ω'/Ωi are calculated for the basic azimuth mode m = 1 (ω' = ω-mωi+, ω+ = (-ωci+Ωi)/2, Ωi = (ωci2-4eEr/mir)1/2 is called the "modified" ion cyclotron (MIC) frequency), for the density fraction of ions of atomic nitrogen f = Ni/ne = 0,01 and are presented in graphic form versus parameter 2ωpe2/ωce2. The spectra of oscillations ω'/Ωi consist of the family of electron Trivel-piece—Gould (TG) modes and of the families of MIC modes. The frequencies of MIC modes are located in a small vicinity of harmonics of the MIC frequency Ωi above and below the harmonic. The TG modes in non-neutral plasma fall in the region of MIC frequencies Ωi and interact strongly with MIC modes. The slow TG modes become unstable near the crossings with non-negative harmonics of MIC frequencies. The instabilities have a resonant character. The lowest radial TG mode has a maximum growth rate at crossing with a zero harmonic of Ωi ((Im ω'/Ωi)max≈0,074). The growth rates of MIC modes are much lower ((Im ω'/Ωi)max≲0,002). Their instability has a threshold character. The instabilities of TG and MIC modes take place mainly at the values of parameter 2ωpe2/ωce2, corresponding to strong radial electric fields (ωci2≪|eEr/mir|), in which the ions are unmagnetized. The oscillations of small amplitude are seen on some frequency

  15. Molecular Design, Structural Analysis and Antifungal Activity of Derivatives of Peptide CGA-N46.

    PubMed

    Li, Rui-Fang; Lu, Zhi-Fang; Sun, Ya-Nan; Chen, Shi-Hua; Yi, Yan-Jie; Zhang, Hui-Ru; Yang, Shuo-Ye; Yu, Guang-Hai; Huang, Liang; Li, Chao-Nan

    2016-09-01

    Chromogranin A (CGA)-N46, a derived peptide of human chromogranin A, has antifungal activity. To further research the active domain of CGA-N46, a series of derivatives were designed by successively deleting amino acid from both terminus of CGA-N46, and the amino acid sequence of each derivative was analyzed by bioinformatic software. Based on the predicted physicochemical properties of the peptides, including half-life time in mammalian reticulocytes (in vitro), yeast (in vivo) and E. coli (in vivo), instability index, aliphatic index and grand average of hydropathicity (GRAVY), the secondary structure, net charge, the distribution of hydrophobic residues and hydrophilic residues, the final derivatives CGA-N15, CGA-N16, CGA-N12 and CGA-N8 were synthesized by solid-phase peptide synthesis. The results of bioinformatic analysis showed that CGA-N46 and its derivatives were α-helix, neutral or weak positive charge, hydrophilic, and CGA-N12 and CGA-N8 were more stable than the other derivatives. The results of circular dichroism confirmed that CGA-N46 and its derived peptides displayed α-helical structure in an aqueous solution and 30 mM sodium dodecylsulfate, but α-helical contents decreased in hydrophobic lipid vesicles. CGA-N15, CGA-N16, CGA-N12 and CGA-N8 had higher antifungal activities than their mother peptide CGA-N46. Among of the derived peptides, CGA-N12 showed the least hemolytic activity. In conclusion, we have successfully identified the active domain of CGA-N46 with strong antifungal activity and weak hemolytic activity, which provides the possibility to develop a new class of antibiotics.

  16. Knowledge and institutional requirements to promote land degradation neutrality in drylands - An analysis of the outcomes of the 3rd UNCCD scientific conference

    NASA Astrophysics Data System (ADS)

    Akhtar-Schuster, Mariam; Safriel, Uriel; Abraham, Elena; de Vente, Joris; Essahli, Wafa; Escadafal, Richard; Stringer, Lindsay

    2015-04-01

    Achieving land degradation neutrality (LDN) through sustainable land management (SLM) targets the maintenance or restoration of the productivity of land, and therefore has to include decision-makers, knowledge generators and knowledge holders at the different relevant geographic scales. In order to enhance the implementation of the Convention, the Conference of the Parties (COP) of the United Nations Convention to Combat Desertification therefore decided that each future session of its Committee on Science and Technology (CST) would be organized in a predominantly scientific and technical conference-style format. This contribution will outline the major outcomes of UNCCD's 3rd scientific conference that will be held in Cancún, Mexico, from 9 to 12 March 2015, on addressing desertification, land degradation and drought issues (DLDD) for poverty reduction and sustainable development. The conference follows an exceptional new round table conference format that will allow the various stakeholders to discuss scientific as well as the contribution of traditional knowledge and practices in combating land degradation. This format should provide two-way communication and enable deeper insight into the availability and contribution of all forms of knowledge for achieving LDN through the assessment of: • the vulnerability of lands to DLDD and climate change and the adaptive capacities of socio-ecosystems; • best examples of adapted, knowledge-based practices and technologies; • monitoring and assessment methods to evaluate the effectiveness of adaptation practices and technologies. The outcomes of UNCCD's 3rd scientific conference will serve as a basis for discussing: • contributions of science to diagnose the status of land; • research gaps that need to be addressed to achieve LDN for poverty reduction; • additional institutional requirements to optimally bridge knowledge generation, knowledge maintenance and knowledge implementation at the science

  17. Activation analysis of the compact ignition tokamak

    SciTech Connect

    Selcow, E.C.

    1986-01-01

    The US fusion program has completed the conceptual design of a compact tokamak device that achieves ignition. The high neutron wall loadings associated with this compact deuterium-tritium-burning device indicate that radiation-related issues may be significant considerations in the overall system design. Sufficient shielding will be requied for the radiation protection of both reactor components and occupational personnel. A close-in igloo shield has been designed around the periphery of the tokamak structure to permit personnel access into the test cell after shutdown and limit the total activation of the test cell components. This paper describes the conceptual design of the igloo shield system and discusses the major neutronic concerns related to the design of the Compact Ignition Tokamak.

  18. Fast Neutral Pressure Gauges in NSTX

    SciTech Connect

    R. Raman; H.W. Kugel; R. Gernhardt; T. Provost; T.R. Jarboe; V. Soukhanovskii

    2004-04-26

    Successful operation in NSTX of two prototype fast-response micro ionization gauges during plasma operations has motivated us to install five gauges at different toroidal and poloidal locations to measure the edge neutral pressure and its dependence on the type of discharge (L-mode, H-mode, CHI) and the fueling method and location. The edge neutral pressure is also used as an input to the transport analysis codes TRANSP and DEGAS-2. The modified PDX-type Penning gauges are well suited for pressure measurements in the NSTX divertor where the toroidal field is relatively high. Behind the NSTX outer divertor plates where the field is lower, an unshielded fast ion gauge of a new design has been installed. This gauge was developed after laboratory testing of several different designs in a vacuum chamber with applied magnetic fields.

  19. Low-energy neutral-atom spectrometer

    SciTech Connect

    Voss, D.E.; Cohen, S.A.

    1982-04-01

    The design, calibration, and performance of a low energy neutral atom spectrometer are described. Time-of-flight analysis is used to measure the energy spectrum of charge-exchange deuterium atoms emitted from the PLT tokamak plasma in the energy range from 20 to 1000 eV. The neutral outflux is gated on a 1 ..mu..sec time scale by a slotted rotating chopper disc, supported against gravity in vacuum by magnetic levitation, and is detected by secondary electron emission from a Cu-Be plate. The energy dependent detection efficiency has been measured in particle beam experiments and on the tokamak so that the diagnostic is absolutely calibrated, allowing quantitative particle fluxes to be determined with 200 ..mu..sec time resolution. In addition to its present application as a plasma diagnostic, the instrument is capable of making a wide variety of measurements relevant to atomic and surface physics.

  20. Phosphorus Determination by Derivative Activation Analysis: A Multifaceted Radiochemical Application.

    ERIC Educational Resources Information Center

    Kleppinger, E. W.; And Others

    1984-01-01

    Although determination of phosphorus is important in biology, physiology, and environmental science, traditional gravimetric and colorimetric methods are cumbersome and lack the requisite sensitivity. Therefore, a derivative activation analysis method is suggested. Background information, procedures, and results are provided. (JN)

  1. Human cytomegalovirus neutralizing antibody-resistant phenotype is associated with reduced expression of glycoprotein H.

    PubMed Central

    Li, L; Coelingh, K L; Britt, W J

    1995-01-01

    We have characterized a neutralizing antibody-resistant mutant human cytomegalovirus (HCMV) obtained from a patient treated with a human monoclonal antiglycoprotein H (gH; unique long region 75) antibody. This virus exhibited resistance to several different neutralizing anti-gH murine monoclonal antibodies (MAbs), as well as to a polyvalent anti-gH serum. The resistant phenotype was unstable and could be maintained only by passage of plaque-purified virus under neutralizing MAb selection. In the absence of a MAb, the resistant phenotype reverted to a neutralizing antibody-sensitive phenotype within one passage. The predicted amino acid sequences of gH from the MAb-resistant and -susceptible parent viruses were identical. Biochemical analysis of the MAb-resistant and -susceptible parent viruses revealed a marked decrease of gH expression in the envelope of the MAb-resistant virus. Furthermore, propagation of the virus in various MAb concentrations resulted in the production of extracellular virions with various levels of resistance to the neutralizing activity of the MAb. These results suggest a mechanism for the generation of neutralizing antibody-resistant viruses which could evade host-derived antiviral antibody responses. In addition, our findings indicate that the stoichiometry of gH in the envelope of infectious HCMV virions is not rigidly fixed and therefore offer a simple explanation for production of phenotypic variants of HCMV through an assembly process in which the content of gH in the envelope of progeny virions varies randomly. PMID:7666509

  2. TPX/TFTR Neutral Beam energy absorbers

    SciTech Connect

    Dahlgren, F.; Wright, K.; Kamperschroer, J.; Grisham, L.; Lontai, L.; Peters, C.; VonHalle, A.

    1993-11-01

    The present beam energy absorbing surfaces on the TFTR Neutral Beams such as Ion Dumps, Calorimeters, beam defining apertures, and scrapers, are simple water cooled copper plates which wee designed to absorb (via their thermal inertia) the incident beam power for two seconds with a five minute coal down interval between pulses. These components are not capable of absorbing the anticipated beam power loading for 1000 second TPX pulses and will have to be replaced with an actively cooled design. While several actively cooled energy absorbing designs were considered,, the hypervapotron elements currently being used on the JET beamlines were chosen due to their lower cooling water demands and reliable performance on JET.

  3. Expression of apical Na(+)-L-glutamine co-transport activity, B(0)-system neutral amino acid co-transporter (B(0)AT1) and angiotensin-converting enzyme 2 along the jejunal crypt-villus axis in young pigs fed a liquid formula

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Gut apical amino acid (AA) transport activity is high at birth and during suckling, thus being essential to maintain luminal nutrient-dependent mucosal growth through providing AA as essential metabolic fuel, substrates and nutrient stimuli for cellular growth. Because system-B(0) Na(+)-neutral AA c...

  4. Neutral Supersymmetric Higgs Boson Searches

    SciTech Connect

    Robinson, Stephen Luke

    2008-07-01

    In some Supersymmetric extensions of the Standard Model, including the Minimal Supersymmetric Standard Model (MSSM), the coupling of Higgs bosons to b-quarks is enhanced. This enhancement makes the associated production of the Higgs with b-quarks an interesting search channel for the Higgs and Supersymmetry at D0. The identification of b-quarks, both online and offline, is essential to this search effort. This thesis describes the author's involvement in the development of both types of b-tagging and in the application of these techniques to the MSSM Higgs search. Work was carried out on the Level-3 trigger b-tagging algorithms. The impact parameter (IP) b-tagger was retuned and the effects of increased instantaneous luminosity on the tagger were studied. An extension of the IP-tagger to use the z-tracking information was developed. A new b-tagger using secondary vertices was developed and commissioned. A tool was developed to allow the use of large multi-run samples for trigger studies involving b-quarks. Offline, a neural network (NN) b-tagger was trained combining the existing offline lifetime based b-tagging tools. The efficiency and fake rate of the NN b-tagger were measured in data and MC. This b-tagger was internally reviewed and certified by the Collaboration and now provides the official b-tagging for all analyses using the Run IIa dataset at D0. A search was performed for neutral MSSM Higgs bosons decaying to a b{bar b} pair and produced in association with one or more b-quarks. Limits are set on the cross-section times the branching ratio for such a process. The limits were interpreted in various MSSM scenarios. This analysis uses the NN b-tagger and was the first to use this tool. The analysis also relies on triggers using the Level-3 IP b-tagging tool described previously. A likelihood discriminant was used to improve the analysis and a neural network was developed to cross-check this technique. The result of the analysis has been submitted to PRL and

  5. Prompt Gamma-Ray Neutron Activation Analysis (PGNAA) for Elemental Analysis

    SciTech Connect

    Robin P. Gardner

    2006-04-11

    This research project was to improve the prompt gamma-ray neutron activation analysis (PGNAA) measurement approach for bulk analysis, oil well logging, and small sample thermal enutron bean applications.

  6. Analysis of body calcium (regional changes in body calcium by in vivo neutron activation analysis)

    NASA Technical Reports Server (NTRS)

    Suki, W.; Johnson, P. C.; Leblanc, A.; Evans, H. J.

    1981-01-01

    The effect of space flight on urine and fecal calcium loss was documented during the three long-term Skylab flights. Neutron activation analysis was used to determine regional calcium loss. Various designs for regional analysis were investigated.

  7. Analysis of supercooling activities of surfactants.

    PubMed

    Kuwabara, Chikako; Terauchi, Ryuji; Tochigi, Hiroshi; Takaoka, Hisao; Arakawa, Keita; Fujikawa, Seizo

    2014-08-01

    Supercooling-promoting activities (SCAs) of 25 kinds of surfactants including non-ionic, anionic, cationic and amphoteric types were examined in solutions (buffered Milli-Q water, BMQW) containing the ice nucleation bacterium (INB) Erwinia ananas, silver iodide (AgI) or BMQW alone, which unintentionally contained unidentified ice nucleators, by a droplet freezing assay. Most of the surfactants exhibited SCA in solutions containing AgI but not in solutions containing the INB E. ananas or BMQW alone. SCAs of many surfactants in solutions containing AgI were very high compared with those of previously reported supercooling-promoting substances. Cationic surfactants, hexadecyltrimethylammonium bromide (C16TAB) and hexadecyltrimethylammonium chloride (C16TAC), at concentrations of 0.01% (w/v) exhibited SCA of 11.8 °C, which is the highest SCA so far reported. These surfactants also showed high SCAs at very low concentrations in solutions containing AgI. C16TAB exhibited SCA of 5.7 °C at a concentration of 0.0005% (w/v). PMID:24792543

  8. Analysis of supercooling activities of surfactants.

    PubMed

    Kuwabara, Chikako; Terauchi, Ryuji; Tochigi, Hiroshi; Takaoka, Hisao; Arakawa, Keita; Fujikawa, Seizo

    2014-08-01

    Supercooling-promoting activities (SCAs) of 25 kinds of surfactants including non-ionic, anionic, cationic and amphoteric types were examined in solutions (buffered Milli-Q water, BMQW) containing the ice nucleation bacterium (INB) Erwinia ananas, silver iodide (AgI) or BMQW alone, which unintentionally contained unidentified ice nucleators, by a droplet freezing assay. Most of the surfactants exhibited SCA in solutions containing AgI but not in solutions containing the INB E. ananas or BMQW alone. SCAs of many surfactants in solutions containing AgI were very high compared with those of previously reported supercooling-promoting substances. Cationic surfactants, hexadecyltrimethylammonium bromide (C16TAB) and hexadecyltrimethylammonium chloride (C16TAC), at concentrations of 0.01% (w/v) exhibited SCA of 11.8 °C, which is the highest SCA so far reported. These surfactants also showed high SCAs at very low concentrations in solutions containing AgI. C16TAB exhibited SCA of 5.7 °C at a concentration of 0.0005% (w/v).

  9. Surface conversion techniques for low energy neutral atom imagers

    NASA Technical Reports Server (NTRS)

    Quinn, J. M.

    1995-01-01

    This investigation has focused on development of key technology elements for low energy neutral atom imaging. More specifically, we have investigated the conversion of low energy neutral atoms to negatively charged ions upon reflection from specially prepared surfaces. This 'surface conversion' technique appears to offer a unique capability of detecting, and thus imaging, neutral atoms at energies of 0.01 - 1 keV with high enough efficiencies to make practical its application to low energy neutral atom imaging in space. Such imaging offers the opportunity to obtain the first instantaneous global maps of macroscopic plasma features and their temporal variation. Through previous in situ plasma measurements, we have a statistical picture of large scale morphology and local measurements of dynamic processes. However, with in situ techniques it is impossible to characterize or understand many of the global plasma transport and energization processes. A series of global plasma images would greatly advance our understanding of these processes and would provide the context for interpreting previous and future in situ measurements. Fast neutral atoms, created from ions that are neutralized in collisions with exospheric neutrals, offer the means for remotely imaging plasma populations. Energy and mass analysis of these neutrals provides critical information about the source plasma distribution. The flux of neutral atoms available for imaging depends upon a convolution of the ambient plasma distribution with the charge exchange cross section for the background neutral population. Some of the highest signals are at relatively low energies (well below 1 keV). This energy range also includes some of the most important plasma populations to be imaged, for example the base of the cleft ion fountain.

  10. Overview af MSFC's Applied Fluid Dynamics Analysis Group Activities

    NASA Technical Reports Server (NTRS)

    Garcia, Roberto; Griffin, Lisa; Williams, Robert

    2004-01-01

    This paper presents viewgraphs on NASA Marshall Space Flight Center's Applied Fluid Dynamics Analysis Group Activities. The topics include: 1) Status of programs at MSFC; 2) Fluid Mechanics at MSFC; 3) Relevant Fluid Dynamics Activities at MSFC; and 4) Shuttle Return to Flight.

  11. Spatiotemporal Data Mining, Analysis, and Visualization of Human Activity Data

    ERIC Educational Resources Information Center

    Li, Xun

    2012-01-01

    This dissertation addresses the research challenge of developing efficient new methods for discovering useful patterns and knowledge in large volumes of electronically collected spatiotemporal activity data. I propose to analyze three types of such spatiotemporal activity data in a methodological framework that integrates spatial analysis, data…

  12. Understanding Tensions: Activity Systems Analysis of Cross-Continental Collaboration

    ERIC Educational Resources Information Center

    Ryder, LanHui Zhang; Yamagata-Lynch, Lisa

    2014-01-01

    Using the lens of Vygotsky's sociocultural theory, activity theory, and Engeström's activity systems analysis, this qualitative study explores students' experiences in the context of a sixteen-week transpacific collaboration between seven students at Northern Illinois University (NIU) and seven students from Shandong Normal…

  13. Project-Based Language Learning: An Activity Theory Analysis

    ERIC Educational Resources Information Center

    Gibbes, Marina; Carson, Lorna

    2014-01-01

    This paper reports on an investigation of project-based language learning (PBLL) in a university language programme. Learner reflections of project work were analysed through Activity Theory, where tool-mediated activity is understood as the central unit of analysis for human interaction. Data were categorised according to the components of human…

  14. Neutral and Non-Neutral Evolution of Drosophila Mitochondrial DNA

    PubMed Central

    Rand, D. M.; Dorfsman, M.; Kann, L. M.

    1994-01-01

    To test hypotheses of neutral evolution of mitochondrial DNA (mtDNA), nucleotide sequences were determined for 1515 base pairs of the NADH dehydrogenase subunit 5 (ND5) gene in the mitochondrial DNA of 29 lines of Drosophila melanogaster and 9 lines of its sibling species Drosophila simulans. In contrast to the patterns for nuclear genes, where D. melanogaster generally exhibits much less nucleotide polymorphism, the number of segregating sites was slightly higher in a global sample of nine ND5 sequences in D. melanogaster (s = 8) than in the nine lines of D. simulans (s = 6). When compared to variation at nuclear loci, the mtDNA variation in D. melanogaster does not depart from neutral expectations. The ND5 sequences in D. simulans, however, show fewer than half the number of variable sites expected under neutrality when compared to sequences from the period locus. While this reduction in variation is not significant at the 5% level, HKA tests with published restriction data for mtDNA in D. simulans do show a significant reduction of variation suggesting a selective sweep of variation in the mtDNA in this species. Tests of neutral evolution based on the ratios of synonymous and replacement polymorphism and divergence are generally consistent with neutral expectations, although a significant excess of amino acid polymorphism within both species is localized in one region of the protein. The rate of mtDNA evolution has been faster in D. melanogaster than in D. simulans and the population structure of mtDNA is distinct in these species. The data reveal how different rates of mtDNA evolution between species and different histories of neutral and adaptive evolution within species can compromise historical inferences in population and evolutionary biology. PMID:7851771

  15. Potent neutralizing monoclonal antibodies against Ebola virus infection

    PubMed Central

    Zhang, Qi; Gui, Miao; Niu, Xuefeng; He, Shihua; Wang, Ruoke; Feng, Yupeng; Kroeker, Andrea; Zuo, Yanan; Wang, Hua; Wang, Ying; Li, Jiade; Li, Chufang; Shi, Yi; Shi, Xuanling; Gao, George F.; Xiang, Ye; Qiu, Xiangguo; Chen, Ling; Zhang, Linqi

    2016-01-01

    Ebola virus infections cause a deadly hemorrhagic disease for which no vaccines or therapeutics has received regulatory approval. Here we show isolation of three (Q206, Q314 and Q411) neutralizing monoclonal antibodies (mAbs) against the surface glycoprotein (GP) of Ebola virus identified in West Africa in 2014 through sequential immunization of Chinese rhesus macaques and antigen-specific single B cell sorting. These mAbs demonstrated potent neutralizing activities against both pseudo and live Ebola virus independent of complement. Biochemical, single particle EM, and mutagenesis analysis suggested Q206 and Q411 recognized novel epitopes in the head while Q314 targeted the glycan cap in the GP1 subunit. Q206 and Q411 appeared to influence GP binding to its receptor NPC1. Treatment with these mAbs provided partial but significant protection against disease in a mouse model of Ebola virus infection. These novel mAbs could serve as promising candidates for prophylactic and therapeutic interventions against Ebola virus infection. PMID:27181584

  16. Spatial heterogeneity analysis of brain activation in fMRI

    PubMed Central

    Gupta, Lalit; Besseling, René M.H.; Overvliet, Geke M.; Hofman, Paul A.M.; de Louw, Anton; Vaessen, Maarten J.; Aldenkamp, Albert P.; Ulman, Shrutin; Jansen, Jacobus F.A.; Backes, Walter H.

    2014-01-01

    In many brain diseases it can be qualitatively observed that spatial patterns in blood oxygenation level dependent (BOLD) activation maps appear more (diffusively) distributed than in healthy controls. However, measures that can quantitatively characterize this spatial distributiveness in individual subjects are lacking. In this study, we propose a number of spatial heterogeneity measures to characterize brain activation maps. The proposed methods focus on different aspects of heterogeneity, including the shape (compactness), complexity in the distribution of activated regions (fractal dimension and co-occurrence matrix), and gappiness between activated regions (lacunarity). To this end, functional MRI derived activation maps of a language and a motor task were obtained in language impaired children with (Rolandic) epilepsy and compared to age-matched healthy controls. Group analysis of the activation maps revealed no significant differences between patients and controls for both tasks. However, for the language task the activation maps in patients appeared more heterogeneous than in controls. Lacunarity was the best measure to discriminate activation patterns of patients from controls (sensitivity 74%, specificity 70%) and illustrates the increased irregularity of gaps between activated regions in patients. The combination of heterogeneity measures and a support vector machine approach yielded further increase in sensitivity and specificity to 78% and 80%, respectively. This illustrates that activation distributions in impaired brains can be complex and more heterogeneous than in normal brains and cannot be captured fully by a single quantity. In conclusion, heterogeneity analysis has potential to robustly characterize the increased distributiveness of brain activation in individual patients. PMID:25161893

  17. Neutral recirculation—the key to control of divertor operation

    NASA Astrophysics Data System (ADS)

    Kukushkin, A. S.; Pacher, H. D.

    2016-12-01

    Interaction of the plasma with neutral gas in the divertor affects virtually all aspects of divertor functionality (power loading of the targets, pumping and fuelling, sustaining the operational conditions of the core plasma). In the course of ITER design development, this interaction has been the subject of intense modelling analysis, supported by experiments on various tokamaks. Neutral gas puffing is found to be the most effective means of divertor control. The results of those studies are summarized and assessed in the paper.

  18. Neutron activation analysis for antimetabolites. [in food samples

    NASA Technical Reports Server (NTRS)

    1973-01-01

    Determination of metal ion contaminants in food samples is studied. A weighed quantity of each sample was digested in a concentrated mixture of nitric, hydrochloric and perchloric acids to affect complete solution of the food products. The samples were diluted with water and the pH adjusted according to the specific analysis performed. The samples were analyzed by neutron activation analysis, polarography, and atomic absorption spectrophotometry. The solid food samples were also analyzed by neutron activation analysis for increased sensitivity and lower levels of detectability. The results are presented in tabular form.

  19. Purification and characterization of neutral and alkaline invertase from carrot.

    PubMed Central

    Lee, H S; Sturm, A

    1996-01-01

    Neutral and alkaline invertase were identified in cells of a suspension culture of carrot (Daucus carota L.) and purified to electrophoretic homogeneity. Neutral invertase is an octamer with a molecular mass of 456 kD and subunits of 57 kD, whereas alkaline invertase is a tetramer with a molecular mass of 504 kD and subunits of 126 kD. Both enzymes had sharp pH profiles, with maximal activities at pH 6.8 for neutral invertase and pH 8.0 for alkaline invertase, and both hydrolyzed sucrose with typical hyperbolic kinetics and similar Km values of about 20 mM at pH 7.5. Neutral invertase also hydrolyzed raffinose and stachyose and, therefore, is a beta-fructofuranosidase. In contrast, alkaline invertase was highly specific for sucrose. Fructose acted as a competitive inhibitor of both enzymes, with Ki values of about 15 mM. Glucose was a noncompetitive inhibitor of both neutral and alkaline invertase, with a Ki of about 30 mM. Neither enzyme was inhibited by HgCl2. Alkaline invertase was markedly inhibited by CaCl2, MgCl2, and MnCl2, and neutral invertase was not. In contrast to alkaline invertase, neutral invertase was inhibited by the nucleotides ATP, CTP, GTP, and UTP. PMID:8972597

  20. Neutralization of HIV by Milk Expressed Antibody

    PubMed Central

    Yu, Xiaocong; Pollock, Daniel; Duval, Mark; Lewis, Christopher; Joseph, Kristin; Meade, Harry; Cavacini, Lisa

    2012-01-01

    Background In some areas of the world mother-to-child transmission of HIV remains a significant problem in part due to widespread breastfeeding which is essential due to scarce supply of a safe replacement, protection conferred by breast milk against many enteric illnesses, and cultural norms. We propose that sustained, adequate levels of protective antibodies in breast milk will prevent transmission of HIV. Methods The HIV neutralizing human monoclonal antibody b12 (IgG1) has been expressed as an IgA2 in CHO cells and shown to retain full immunoreactivity and neutralizing activity as the parental IgG1. The expression plasmids containing the b12 heavy and light chains were also used to construct milk specific expression vectors using the GTC goat β-casein expression vector to direct expression of linked genes to the mammary gland with subsequent secretion into the milk. Female transgenic mice were generated and following parturition, their milk was tested for antibody immunoreactivity with gp120 and neutralization of HIV. Results When compared to CHO derived b12 IgA2 (or IgG1), immunoreactivity was retained. When tested for neutralization, milk derived b12 IgA2 was at least comparable to CHO derived antibody and in some cases superior to CHO derived antibody. Furthermore, milk that expressed b12 IgA2 was significantly more effective at mediating antibody dependent cell killing. Conclusions These results suggest it is possible to achieve functional HIV-specific mAb in the milk of transgenic mice and further investigations are warranted to explore ways for inducing this type of antibody response in the breast milk of HIV infected women. PMID:23269241