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Analysis of Non-Enzymatically Glycated Peptides: Neutral-Loss Triggered MS3 Versus Multi-Stage Activation Tandem Mass Spectrometry
Zhang, Qibin; Petyuk, Vladislav A.; Schepmoes, Athena A.; Orton, Daniel J.; Monroe, Matthew E.; Yang, Feng; Smith, Richard D.; Metz, Thomas O.
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.
Enterovirus neutralizing activity in the gastrointestinal tract of piglets.
Derbyshire, J B
Neutralizing activity against porcine enterovirus strain T80 was demonstrated in the contents of the stomach, duodenum or ileum of four piglets which were suckling dams whose milk contained neutralizing substances against the same virus. No neutralizing activity was detected in the gastrointestinal contents of an unsuckled piglet or in four weaned piglets. Extracts of intestinal tissue from each of the above piglets failed to neutralize the virus. Four weaned piglets were dosed orally with live T80 virus. From nine days after infection virus neutralizing activity was found in extracts of tissue prepared from the duodenum, jejunum, ileum, caecum and colon but not in the contents of the gastrointestinal tract and a serological response to the virus was demonstrated. No virus neutralizing activity was detected in gastrointestinal tissue or contents from four weaned piglets inoculated parenterally with live T80 virus or in four piglets dosed orally with inactivated T80 virus and these piglets did not respond serologically to the virus.
Computational Prediction of Broadly Neutralizing HIV-1 Antibody Epitopes from Neutralization Activity Data
Ferguson, Andrew L.; Falkowska, Emilia; Walker, Laura M.; Seaman, Michael S.; Burton, Dennis R.; Chakraborty, Arup K.
Broadly neutralizing monoclonal antibodies effective against the majority of circulating isolates of HIV-1 have been isolated from a small number of infected individuals. Definition of the conformational epitopes on the HIV spike to which these antibodies bind is of great value in defining targets for vaccine and drug design. Drawing on techniques from compressed sensing and information theory, we developed a computational methodology to predict key residues constituting the conformational epitopes on the viral spike from cross-clade neutralization activity data. Our approach does not require the availability of structural information for either the antibody or antigen. Predictions of the conformational epitopes of ten broadly neutralizing HIV-1 antibodies are shown to be in good agreement with new and existing experimental data. Our findings suggest that our approach offers a means to accelerate epitope identification for diverse pathogenic antigens. PMID:24312481
Thermochemical Analysis of Neutralization Reactions: An Introductory Discovery Experiment
ERIC Educational Resources Information Center
Mills, Kenneth V.; Gullmette, Louise W.
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.
Seismic active control by neutral networks
Tang, Yu
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.
Park, Jeewoo; Kucharek, Harald; Möbius, Eberhard; Galli, André; Livadiotis, George; Fuselier, Steve A.; McComas, David J.
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.
Statistical Analysis of the Heavy Neutral Atoms Measured by IBEX
NASA Astrophysics Data System (ADS)
Park, Jeewoo; Kucharek, Harald; Möbius, Eberhard; Galli, André; Livadiotis, George; Fuselier, Steve A.; McComas, David J.
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&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&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.
Launch Deployment Assembly Extravehicular Activity Neutral Buoyancy Development Test Report
NASA Technical Reports Server (NTRS)
Loughead, T.
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).
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.
, yielding the necessary ion velocity spectra needed to further analysis of parent neutral exosphere properties.
Mitigating the antimicrobial activities of selected organic acids and commercial sanitizers with various neutralizing agents.
Park, Yoen Ju; Chen, Jinru
This study was conducted to evaluate the abilities of five neutralizing agents, Dey-Engley (DE) neutralizing broth (single or double strength), morpholinepropanesulfonic acid (MOPS) buffer, phosphate-buffered saline (PBS), and sodium thiosulfate buffer, in mitigating the activities of acetic or lactic acid (2%) and an alkaline or acidic sanitizer (a manufacturer-recommended concentration) againt the cells of Shiga toxin-producing Escherichia coli (STEC; n = 9). To evaluate the possible toxicity of the neutralizing agents to the STEC cells, each STEC strain was exposed to each of the neutralizing agents at room temperature for 10 min. Neutralizing efficacy was evaluated by placing each STEC strain in a mixture of sanitizer and neutralizer under the same conditions. The neutralizing agents had no detectable toxic effect on the STEC strains. PBS was least effective for neutralizing the activity of selected organic acids and sanitizers. Single-strength DE and sodium thiosulfate neutralized the activity of both acetic and lactic acids. MOPS buffer neutralized the activity of acetic acid and lactic acid against six and five STEC strains, respectively. All neutralizing agents, except double-strength DE broth, had a limited neutralizing effect on the activity of the commercial sanitizers used in the study. The double-strength DE broth effectively neutralized the activity of the two commercial sanitizers with no detectable toxic effects on STEC cells.
Neutral density filters with Risley prisms: analysis and design.
Duma, Virgil-Florin; Nicolov, Mirela
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.
Improving the neutral phytase activity from Bacillus amyloliquefaciens DSM 1061 by site-directed mutagenesis.
Xu, Wei; Shao, Rong; Wang, Zupeng; Yan, Xiuhua
Neutral phytase is used as a feed additive for degradation of anti-nutritional phytate in aquatic feed industry. Site-directed mutagenesis of Bacillus amyloliquefaciens DSM 1061 phytase was performed with an aim to increase its activity. Mutation residues were chosen based on multiple sequence alignments and structure analysis of neutral phytsaes from different microorganisms. The mutation sites on surface (D148E, S197E and N156E) and around the active site (D52E) of phytase were selected. Analysis of the phytase variants showed that the specific activities of mutants D148E and S197E remarkably increased by about 35 and 13% over a temperature range of 40-75 °C at pH 7.0, respectively. The k cat of mutants D148E and S197E were 1.50 and 1.25 times than that of the wild-type phytase, respectively. Both D148E and S197E showed much higher thermostability than that of the wild-type phytase. However, mutants N156E and D52E led to significant loss of specific activity of the enzyme. Structural analysis revealed that these mutations may affect conformation of the active site of phytase. The present mutant phytases D148E and S197E with increased activities and thermostabilities have application potential as additives in aquaculture feed.
Magnitudes of muscle activation of spine stabilizers, gluteals, and hamstrings during supine bridge to neutral position.
Youdas, James W; Hartman, James P; Murphy, Brooke A; Rundle, Ashley M; Ugorowski, Jenna M; Hollman, John H
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.
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
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).
In Vitro Selection of Cathepsin E-Activity-Enhancing Peptide Aptamers at Neutral pH
Biyani, Madhu; Futakami, Masae; Kitamura, Koichiro; Kawakubo, Tomoyo; Suzuki, Miho; Yamamoto, Kenji; Nishigaki, Koichi
The aspartic protease cathepsin E has been shown to induce apoptosis in cancer cells under physiological conditions. Therefore, cathepsin E-activity-enhancing peptides functioning in the physiological pH range are valuable potential cancer therapeutic candidates. Here, we have used a general in vitro selection method (evolutionary rapid panning analysis system (eRAPANSY)), based on inverse substrate-function link (SF-link) selection to successfully identify cathepsin E-activity-enhancing peptide aptamers at neutral pH. A successive enrichment of peptide activators was attained in the course of selection. One such peptide activated cathepsin E up to 260%, had a high affinity (KD; ∼300 nM), and had physiological activity as demonstrated by its apoptosis-inducing reaction in cancerous cells. This method is expected to be widely applicable for the identification of protease-activity-enhancing peptide aptamers. PMID:21527983
Au@Ag Heterogeneous Nanorods as Nanozyme Interfaces with Peroxidase-Like Activity and Their Application for One-Pot Analysis of Glucose at Nearly Neutral pH.
Han, Lei; Li, Cuncheng; Zhang, Tao; Lang, Qiaolin; Liu, Aihua
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.
Neutralization of Apis mellifera bee venom activities by suramin.
El-Kik, Camila Z; Fernandes, Fabrício F A; Tomaz, Marcelo Amorim; Gaban, Glauco A; Fonseca, Tatiane F; Calil-Elias, Sabrina; Oliveira, Suellen D S; Silva, Claudia L M; Martinez, Ana Maria Blanco; Melo, Paulo A
In this work we evaluated the ability of suramin, a polysulfonated naphthylurea derivative, to antagonize the cytotoxic and enzymatic effects of the crude venom of Apis mellifera. Suramin was efficient to decrease the lethality in a dose-dependent way. The hemoconcentration caused by lethal dose injection of bee venom was abolished by suramin (30 μg/g). The edematogenic activity of the venom (0.3 μg/g) was antagonized by suramin (10 μg/g) in all treatment protocols. The changes in the vascular permeability caused by A. mellifera (1 μg/g) venom were inhibited by suramin (30 μg/g) in the pre- and posttreatment as well as when the venom was preincubated with suramin. In addition, suramin also inhibited cultured endothelial cell lesion, as well as in vitro myotoxicity, evaluated in mouse extensor digitorum longus muscle, which was inhibited by suramin (10 and 25 μM), decreasing the rate of CK release, showing that suramin protected the sarcolemma against damage induced by components of bee venom (2.5 μg/mL). Moreover, suramin inhibited the in vivo myotoxicity induced by i.m. injection of A. mellifera venom in mice (0.5 μg/g). The analysis of the area under the plasma CK vs. time curve showed that preincubation, pre- and posttreatment with suramin (30 μg/g) inhibited bee venom myotoxic activity in mice by about 89%, 45% and 40%, respectively. Suramin markedly inhibited the PLA2 activity in a concentration-dependent way (1-30 μM). Being suramin a polyanion molecule, the effects observed may be due to the interaction of its charges with the polycation components present in A. mellifera bee venom.
EL4 cell-based colorimetric toxin neutralization activity assays for determination of neutralizing anti-ricin antibodies.
Lindsey, Changhong Y; Brown, J Edward; Torabazar, Nahid R; Smith, Leonard A
A recombinant ricin toxin A-chain 1-33/44-198 vaccine (RVEc), developed at the United States Army Medical Research Institute of Infectious Diseases as a vaccine candidate, is under investigation in a phase 1 clinical study. To effectively evaluate the immunogenicity of this ricin vaccine and to eliminate the use of radioactive material, an EL4 cell-based colorimetric toxin neutralization activity (TNA) assay using a CellTiter 96 AQueous One Solution Cell Proliferation Assay Reagent has been developed, optimized, and applied in the vaccine efficacy studies. The TNA assay measures the protective neutralizing anti-ricin antibodies in animal sera by determining the cell viability after ricin exposure in the assay system and comparing it to a purified mouse polyclonal antiricin IgG standard curve. The standard curve of the anti-ricin TNA assay closely fits a four-parameter logistic regression model. The unknown test sample concentration was expressed as microg/mL, but not the 50% effective concentration (EC50), which was determined by most TNA assays. The neutralizing endpoint titers, not the 50% effective dilution (ED50), of human specimens were measured with the TNA assay in support of the clinical study of the RVEc vaccine. The optimal amount of ricin toxin, EL4 cells, and concentration of standards used in the assay system was established to minimize false-negative and false-positive results of serum specimens from the nonclinical and clinical studies of RVEc. The testing conditions were adjusted to optimize assay performance. The colorimetric TNA assay replaced a radioactive TNA assay previously used in the ricin vaccine studies.
Neutralizing activities against seasonal influenza viruses in human intravenous immunoglobulin
Onodera, Hiroyuki; Urayama, Takeru; Hirota, Kazue; Maeda, Kazuhiro; Kubota-Koketsu, Ritsuko; Takahashi, Kazuo; Hagiwara, Katsuro; Okuno, Yoshinobu; Ikuta, Kazuyoshi; Yunoki, Mikihiro
Influenza viruses A/H1N1, A/H3N2, and B are known seasonal viruses that undergo annual mutation. Intravenous immunoglobulin (IVIG) contains anti-seasonal influenza virus globulins. Although the virus-neutralizing (VN) titer is an indicator of protective antibodies, changes in this titer over extended time periods have yet to be examined. In this study, variations in hemagglutination inhibition (HI) and VN titers against seasonal influenza viruses in IVIG lots over extended time periods were examined. In addition, the importance of monitoring the reactivity of IVIG against seasonal influenza viruses with varying antigenicity was evaluated. A/H1N1, A/H3N2, and B influenza virus strains and IVIG lots manufactured from 1999 to 2014 were examined. The HI titer was measured by standard methods. The VN titer was measured using a micro-focus method. IVIG exhibited significant HI and VN titers against all investigated strains. Our results suggest that the donor population maintains both specific and cross-reactive antibodies against seasonal influenza viruses, except in cases of pandemic viruses, despite major antigen changes. The titers against seasonal influenza vaccine strains, including past strains, were stable over short time periods but increased slowly over time. PMID:28331286
Promoting Active Species Generation by Electrochemical Activation in Alkaline Media for Efficient Electrocatalytic Oxygen Evolution in Neutral Media.
Xu, Kun; Cheng, Han; Liu, Linqi; Lv, Haifeng; Wu, Xiaojun; Wu, Changzheng; Xie, Yi
In this study, by using dicobalt phosphide nanoparticles as precatalysts, we demonstrated that electrochemical activation of metallic precatalysts in alkaline media (comparing with directly electrochemical activation in neutral media) could significantly promote the OER catalysis in neutral media, specifically realizing a 2-fold enhanced activity and meanwhile showing a greatly decreased overpotential of about 100 mV at 10 mA cm(-2). Compared directly with electrochemical activation in neutral media, the electrochemical activation in harsh alkaline media could easily break the strong Co-Co bond and promote active species generation on the surface of metallic Co2P, thus accounting for the enhancement of neutral OER activity, which is also evidenced by HRTEM and the electrochemical double-layer capacitance measurement. The activation of electrochemical oxidation of metallic precatalysts in alkaline media enhanced neutral OER catalysis could also be observed on CoP nanoparticles and Ni2P nanoparticles, suggesting this is a generic strategy. Our work highlights that the activation of electrochemical oxidation of metallic precatalysts in alkaline media would pave new avenues for the design of advanced neutral OER electrocatalysts.
Ion-neutral coupling: Geomagnetic activity modulation of the Equatorial Thermosphere Anomaly
NASA Astrophysics Data System (ADS)
Lei, J.; Dou, X.; Thayer, J. P.; Wang, W.; Luan, X.
The two-way momentum coupling between the neutral thermosphere and its plasma environment plays an important role in producing the trough of the Equatorial Thermosphere Anomaly (ETA). It was found that field-aligned ion drag associated with the equatorial ionosphere anomaly (EIA) can result in vertical motion in the neutral gas over the magnetic equator leading to a reduction in temperature and a trough in thermosphere density at a given altitude. It was also found that the formation of the ETA crests is attributed to plasma-neutral heating which has two peaks in the topside ionosphere aside the magnetic equator. This study is devoted to address the geomagnetic activity affect on the coupling between the equatorial ionosphere and thermosphere on the basis of satellite observations and first-principle ionosphere-thermosphere simulations. The deposited magnetospheric energy into the upper atmosphere associated with geomagnetic activity changes the ionosphere and its background neutral atmosphere significantly and hence their coupling processes. On the other hand, the enhanced electric field and neutral wind during storm time can modulate the EIA and ETA and alter the momentum and energy coupling between the thermosphere and ionosphere in the equatorial F region. Results from NCAR-TIEGCM simulations will help elucidate the ion-neutral coupling processes related to the ETA under active geomagnetic conditions
An analysis of bipropellant neutralization for spacecraft refueling operations
NASA Technical Reports Server (NTRS)
Kauffman, David
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.
Values in the Net Neutrality Debate: Applying Content Analysis to Testimonies from Public Hearings
ERIC Educational Resources Information Center
Cheng, An-Shou
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…
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.
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…
Neutral meson and direct photon analysis with ALICE
NASA Astrophysics Data System (ADS)
Whitehead, A. M.;
The measurement of neutral mesons, particularly π0, s and η's, plays an important role in the study of the Quark-Gluon Plasma (QGP), the hot and dense medium created in high-energy heavy-ion collisions. Parton energy loss in the QGP, often called jet quenching, can be assessed via measuring the suppression of high-pT π0, s in heavy-ion collisions, when compared to pp collisions using the nuclear modification factor (Raa). Furthermore, neutral mesons are the dominant source of photons in pp and Pb-Pb collisions, and their precise measurement is required to measure direct photons that are produced thermally within the QGP or in hard initial scatterings in the earliest phases of the collision. In both cases, high- quality measurements in pp collisions are required as a reference for Pb-Pb collisions. ALICE measurements of neutral meson spectra cover a large p T range, with the Photon Conversion Method - which requires measurements from the ITS and TPC - covering low to intermediate p T and the PHOS and EMCal electromagnetic calorimeters covering an intermediate to high p T range. In this presentation, measurements of π0, s and η's obtained from the ALICE experiment, for pp collisions at several collisional center of mass energies ≤ft(\\sqrt{{s}NN}\\right), from 0.9 TeV to 8 TeV and in Pb-Pb collisions at \\sqrt{{s}NN}=2.76 {{TeV}}, will be presented. The reconstruction of neutral mesons using the Photon Conversion Method (PCM) will also be discussed.
Heavy Neutral Beam Probe for Edge Plasma Analysis in Tokamaks
Castracane, J.
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.
Preclinical evaluation of three polyspecific antivenoms against the venom of Echis ocellatus: Neutralization of toxic activities and antivenomics.
Calvete, Juan J; Arias, Ana Silvia; Rodríguez, Yania; Quesada-Bernat, Sarai; Sánchez, Laura V; Chippaux, Jean Philippe; Pla, Davinia; Gutiérrez, José María
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.
Detection of Broadly Neutralizing Activity within the First Months of HIV-1 Infection
Fabra-Garcia, A.; Gonzalez, N.; Nicolas, D.; Merino-Mansilla, A.; Manzardo, C.; Ambrosioni, J.; Schultz, A.; Meyerhans, A.; Mascola, J. R.; Gatell, J. M.; Alcami, J.; Miro, J. M.; Yuste, E.
ABSTRACT A fraction of HIV-1 patients are able to generate broadly neutralizing antibodies (bNAbs) after 2 to 4 years of infection. In rare occasions such antibodies are observed close to the first year of HIV-1 infection but never within the first 6 months. In this study, we analyzed the neutralization breadth of sera from 157 antiretroviral-naive individuals who were infected for less than 1 year. A range of neutralizing activities was observed with a previously described panel of six recombinant viruses from five different subtypes (M. Medina-Ramirez et al., J Virol 85:5804–5813, 2011, http://dx.doi.org/10.1128/JVI.02482-10). Some sera were broadly reactive, predominantly targeting envelope epitopes within the V2 glycan-dependent region. The neutralization breadth was positively associated with time postinfection (P = 0.0001), but contrary to what has been reported for chronic infections, no association with the viral load was observed. Notably, five individuals within the first 6 months of infection (two as early as 77 and 96 days postinfection) showed substantial cross-neutralization. This was confirmed with an extended panel of 20 Env pseudoviruses from four different subtypes (two in tier 3, 14 in tier 2, and four in tier 1). Sera from these individuals were capable of neutralizing viruses from four different subtypes with a geometric mean 50% infective dose (ID50) between 100 and 800. These results indicate that induction of cross-neutralizing responses, albeit rare, is achievable even within 6 months of HIV-1 infection. These observations encourage the search for immunogens able to elicit this kind of response in preventive HIV-1 vaccine approaches. IMPORTANCE There are very few individuals able to mount broadly neutralizing activity (bNA) close to the first year postinfection. It is not known how early in the infection cross-neutralizing responses can be induced. In the present study, we show that bNAbs, despite being rare, can be induced much earlier
Tissue Plasminogen Activator Neurotoxicity is Neutralized by Recombinant ADAMTS 13
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
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
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.
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
Neutralizing activities of caprine antibodies towards conserved regions of the HCV envelope glycoprotein E2
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
Pseudotype-Based Neutralization Assays for Influenza: A Systematic Analysis
Carnell, George William; Ferrara, Francesca; Grehan, Keith; Thompson, Craig Peter; Temperton, Nigel James
The use of vaccination against the influenza virus remains the most effective method of mitigating the significant morbidity and mortality caused by this virus. Antibodies elicited by currently licensed influenza vaccines are predominantly hemagglutination-inhibition (HI)-competent antibodies that target the globular head of hemagglutinin (HA) thus inhibiting influenza virus entry into target cells. These antibodies predominantly confer homosubtypic/strain specific protection and only rarely confer heterosubtypic protection. However, recent academia or pharma-led R&D toward the production of a "universal vaccine" has centered on the elicitation of antibodies directed against the stalk of the influenza HA that has been shown to confer broad protection across a range of different subtypes (H1–H16). The accurate and sensitive measurement of antibody responses elicited by these "next-generation" influenza vaccines is, however, hampered by the lack of sensitivity of the traditional influenza serological assays HI, single radial hemolysis, and microneutralization. Assays utilizing pseudotypes, chimeric viruses bearing influenza glycoproteins, have been shown to be highly efficient for the measurement of homosubtypic and heterosubtypic broadly neutralizing antibodies, making them ideal serological tools for the study of cross-protective responses against multiple influenza subtypes with pandemic potential. In this review, we will analyze and compare literature involving the production of influenza pseudotypes with particular emphasis on their use in serum antibody neutralization assays. This will enable us to establish the parameters required for optimization and propose a consensus protocol to be employed for the further deployment of these assays in influenza vaccine immunogenicity studies. PMID:25972865
Method of active charge and current neutralization of intense ion beams for ICF
Guiragossian, Z.G.T.; Orthel, J.L.; Lemons, D.S.; Thode, L.E.
Methods of generating the beam neutralization electrons with required properties are given in the context of a Light Ion Fusion Experiment (LIFE) designed accelerator. Recently derived envelope equations for neutralized and ballistically focused intense ion beams are applied to the LIFE geometry in which 10 MeV He/sup +/ multiple beamlets coalesce and undergo 45:1 radial compression while beam pulses experience a 20:1 axial compression in the propagation range of 10 m. Both active and auto-neutralization methods are examined and found to produce initial electron temperatures consistent with the requirement of the envelope equation for both radial and axial adiabatic beam pulse compressions. The stability of neutralized beam propagation is also examined concerning the Pierce type electrostatic instability and for the case of LIFE beams it is found to have insignificant effect. A scaled experimental setup is presented which can serve to perform near term tests on the ballistically focused propagation of neutralized light ion beams.
Standardized, mathematical model-based and validated in vitro analysis of anthrax lethal toxin neutralization.
Li, Han; Soroka, Stephen D; Taylor, Thomas H; Stamey, Karen L; Stinson, Kelly Wallace; Freeman, Alison E; Abramson, Darbi R; Desai, Rita; Cronin, Li X; Oxford, J Wade; Caba, Joseph; Pleatman, Cynthia; Pathak, Sonal; Schmidt, Daniel S; Semenova, Vera A; Martin, Sandra K; Wilkins, Patricia P; Quinn, Conrad P
Quantification of anthrax lethal toxin (LTx) neutralization activity (TNA) is pivotal in assessing protective antibody responses to anthrax vaccines and for evaluation of immunotherapies for anthrax. We have adapted and redesigned the TNA assay to establish a unifying, standardized, quantitative and validated technology platform for LTx neutralization in the J774A.1 murine cell line. Critical design features of this platform are 1) the application of a free-form or constrained 4 parameter logistic (4-PL) function to model neutralization responses within and between boundary limits of 100% cell survival and 95% cell lysis and 2) to exploit innovative assay curve recognition algorithms for interpretive endpoints. The assay was validated using human serum ED50 (dilution of serum effecting 50% neutralization) as the primary reportable value (RV). Intra-operator and intermediate precision, expressed as the coefficient of variation (%CV), were high at 10.5-15.5%CV and 13.5-14.5%CV respectively. TNA assay dilutional linearity was demonstrated for human sera using linear regression analysis of log(10) transformed data with slope=0.99, intercept=-0.03 and r(2)=0.985. Assay accuracy, inferred from the precision and linearity data and using a spike-recovery approach, was high with a percent error (%E) range of only 3.4-20.5%E. The lower limit of detection (LLOD) was ED50=12 and the lower limit of quantification (LLOQ) was ED50=36. The cell-based assay was robust, tolerating incubation temperatures from 35 to 39 degrees C, CO(2) concentrations from 3% to 7% and reporter substrate (MTT) concentrations of 2.5-7.5 mg/ml. Strict assay quality control parameters were met for up to 25 cell culture passages. The long term (50 month) assay stability, determined using human reference standards AVR414 and AVR801, indicated high precision, consistent accuracy and no detectable assay drift. A customized software program provided two additional assay metrics, Quantification Titer (QT) and
Chemical acceleration of a neutral granulated blast-furnace slag activated by sodium carbonate
Kovtun, Maxim Kearsley, Elsabe P. Shekhovtsova, Julia
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.
Potent Intratype Neutralizing Activity Distinguishes Human Immunodeficiency Virus Type 2 (HIV-2) from HIV-1
Özkaya Şahin, Gülşen; Holmgren, Birgitta; da Silva, Zacarias; Nielsen, Jens; Nowroozalizadeh, Salma; Esbjörnsson, Joakim; Månsson, Fredrik; Andersson, Sören; Norrgren, Hans; Aaby, Peter
HIV-2 has a lower pathogenicity and transmission rate than HIV-1. Neutralizing antibodies could be contributing to these observations. Here we explored side by side the potency and breadth of intratype and intertype neutralizing activity (NAc) in plasma of 20 HIV-1-, 20 HIV-2-, and 11 dually HIV-1/2 (HIV-D)-seropositive individuals from Guinea-Bissau, West Africa. Panels of primary isolates, five HIV-1 and five HIV-2 isolates, were tested in a plaque reduction assay using U87.CD4-CCR5 cells as targets. Intratype NAc in HIV-2 plasma was found to be considerably more potent and also broader than intratype NAc in HIV-1 plasma. This indicates that HIV-2-infected individuals display potent type-specific neutralizing antibodies, whereas such strong type-specific antibodies are absent in HIV-1 infection. Furthermore, the potency of intratype NAc was positively associated with the viral load of HIV-1 but not HIV-2, suggesting that NAc in HIV-1 infection is more antigen stimulation dependent than in HIV-2 infection, where plasma viral loads typically are at least 10-fold lower than in HIV-1 infection. Intertype NAc of both HIV-1 and HIV-2 infections was, instead, of low potency. HIV-D subjects had NAc to HIV-2 with similar high potency as singly HIV-2-infected individuals, whereas neutralization of HIV-1 remained poor, indicating that the difference in NAc between HIV-1 and HIV-2 infections depends on the virus itself. We suggest that immunogenicity and/or antigenicity, meaning the neutralization phenotype, of HIV-2 is distinct from that of HIV-1 and that HIV-2 may display structures that favor triggering of potent neutralizing antibody responses. PMID:22072782
Active to sterile neutrino mixing limits from neutral-current interactions in MINOS.
Adamson, P; Auty, D J; Ayres, D S; Backhouse, C; Barr, G; Bishai, M; Blake, A; Bock, G J; Boehnlein, D J; Bogert, D; Cavanaugh, S; Cherdack, D; Childress, S; Coelho, J A B; Coleman, S J; Corwin, L; Cronin-Hennessy, D; Danko, I Z; de Jong, J K; Devenish, N E; Diwan, M V; Dorman, M; Escobar, C O; Evans, J J; Falk, E; Feldman, G J; Frohne, M V; Gallagher, H R; Gomes, R A; Goodman, M C; Gouffon, P; Graf, N; Gran, R; Grant, N; Grzelak, K; Habig, A; Harris, D; Hartnell, J; Hatcher, R; Himmel, A; Holin, A; Huang, X; Hylen, J; Ilic, J; Irwin, G M; Isvan, Z; Jaffe, D E; James, C; Jensen, D; Kafka, T; Kasahara, S M S; Koizumi, G; Kopp, S; Kordosky, M; Kreymer, A; Lang, K; Lefeuvre, G; Ling, J; Litchfield, P J; Loiacono, L; Lucas, P; Mann, W A; Marshak, M L; Mayer, N; McGowan, A M; Mehdiyev, R; Meier, J R; Messier, M D; Miller, W H; Mishra, S R; Mitchell, J; Moore, C D; Morfín, J; Mualem, L; Mufson, S; Musser, J; Naples, D; Nelson, J K; Newman, H B; Nichol, R J; Nicholls, T C; Nowak, J A; Oliver, W P; Orchanian, M; Paley, J; Patterson, R B; Pawloski, G; Pearce, G F; Petyt, D A; Phan-Budd, S; Pittam, R; Plunkett, R K; Qiu, X; Ratchford, J; Raufer, T M; Rebel, B; Rodrigues, P A; Rosenfeld, C; Rubin, H A; Sanchez, M C; Schneps, J; Schreiner, P; Sharma, R; Shanahan, P; Sousa, A; Stamoulis, P; Strait, M; Tagg, N; Talaga, R L; Tetteh-Lartey, E; Thomas, J; Thomson, M A; Tinti, G; Toner, R; Torretta, D; Tzanakos, G; Urheim, J; Vahle, P; Viren, B; Walding, J J; Weber, A; Webb, R C; White, C; Whitehead, L; Wojcicki, S G; Zwaska, R
Results are reported from a search for active to sterile neutrino oscillations in the MINOS long-baseline experiment, based on the observation of neutral-current neutrino interactions, from an exposure to the NuMI neutrino beam of 7.07×10(20) protons on target. A total of 802 neutral-current event candidates is observed in the Far Detector, compared to an expected number of 754 ± 28(stat) ± 37(syst) for oscillations among three active flavors. The fraction f(s) of disappearing ν(μ) that may transition to ν(s) is found to be less than 22% at the 90% C.L.
Activity Coefficient Derivatives of Ternary Systems Based on Scatchard's Neutral Electrolyte description
Miller, D G
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.
Pyrazinamide Is Active against Mycobacterium tuberculosis Cultures at Neutral pH and Low Temperature.
den Hertog, Alice L; Menting, Sandra; Pfeltz, Richard; Warns, Matthew; Siddiqi, Salman H; Anthony, Richard M
For the past decades, an acidic pH has been used to render Mycobacterium tuberculosis susceptible to pyrazinamide for in vitro testing. Here, we show that at the standard breakpoint concentration and reduced culture temperatures, pyrazinamide (PZA) is active against tuberculosis (TB) at neutral pH. This finding should help unravel the mechanism of action of PZA and allow drug susceptibility testing (DST) methods to be optimized.
Studies of a calcium-activated neutral protease from chicken skeletal muscle. I. Purification and characterization.
Ishiura, S; Murofushi, H; Suzuki, K; Imahori, K
A calcium-activated neutral protease was purified 2,700-fold over the crude extract from chicken skeletal muscle. The purified protease migrated as a single band on polyacrylamide gel electrophoresis with or without SDS. Its molecular weight was 80,000 and pH optimum for activity was 7.7. The activity required strictly the presence of calcium (optimum concentration: 1.8 mM) or strontium (optimum concentration: 10 mM) ions. The protease was inhibited by leupeptin, which is known to be a strong inhibitor of papain, cathepsin B, trypsin, and plasmin.
Accumulated Bending Energy Elicits Neutral Sphingomyelinase Activity in Human Red Blood Cells
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
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
Accumulated bending energy elicits neutral sphingomyelinase activity in human red blood cells.
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
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 A(2). 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 A(2) 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.
Alkaline, acid, and neutral phosphatase activities are induced during development in Myxococcus xanthus.
Weinberg, R A; Zusman, D R
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
Classification of illicit heroin by UPLC-Q-TOF analysis of acidic and neutral manufacturing impurities.
Liu, Cuimei; Hua, Zhendong; Bai, Yanping
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.
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
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.
Analysis of wave-like oscillations in parameters of sporadic E layer and neutral atmosphere
NASA Astrophysics Data System (ADS)
Mošna, Z.; Koucká Knížová, P.
The present study mainly concerns the wave-like activity in the ionospheric sporadic E layer (Es) and in the lower lying stratosphere. The proposed analysis involves parameters describing the state of plasma in the sporadic E layer. Critical frequencies foEs and layer heights hEs were measured at the Pruhonice station (50°N, 14.5°E) during summer campaigns 2004, 2006 and 2008. Further, we use neutral atmosphere (temperature data at 10 hPa) data from the same time interval. The analysis concentrates on vertically propagating wave-like structures within distant atmospheric regions. By means of continuous wavelet transform (CWT) we have detected significant wave-like oscillation at periods covering tidal and planetary oscillation domains both in the Es layer parameters (some of them were reported earlier, for instance in works of Abdu et al., 2003; Pancheva and Mitchel, 2004; Pancheva et al., 2003; Šauli and Bourdillon, 2008) and in stratospheric temperature variations. Further analyses using cross wavelet transform (XWT) and wavelet coherence analysis (WTC) show that despite high wave-like activity in a wide period range, there are only limited coherent wave-like bursts present in both spectra. Such common coherent wave bursts occur on periods close to eigen-periods of the terrestrial atmosphere. We suppose that vertical coupling between atmospheric regions realized by vertically propagating planetary waves occurs predominantly on periods close to those of Rossby modes. Analysis of the phase shift between data from distant atmospheric regions reveals high variability and very likely supports the non-linear scenario of the vertical coupling provided by planetary waves.
Kinetics of positive ions and electrically neutral active particles in afterglow in neon at low pressure
Pejović, Milić M. Nešić, Nikola T.; Pejović, Momčilo M.
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.
Activity and stability trends of perovskite oxides for oxygen evolution catalysis at neutral pH.
Han, Binghong; Risch, Marcel; Lee, Yueh-Lin; Ling, Chen; Jia, Hongfei; Shao-Horn, Yang
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.
Adsorption of Neutral Red onto Mn-impregnated activated carbons prepared from Typha orientalis.
Zhang, Jian; Shi, Qianqian; Zhang, Chenglu; Xu, Jingtao; Zhai, Bing; Zhang, Bo
Activated carbon was prepared from an inexpensive and renewable carbon source, Typha orientalis, by H(3)PO(4) activation and then impregnated with different Mn salts and tested for its Neutral Red (NR) adsorption capacities. The amount of Mn impregnated in the activated carbon was influenced by the anion species. Impregnation with Mn decreased the surface area, changed the pore size and crystal structure, and introduced more acidic functional groups such as carboxyl, lactone and phenol groups. The optimum adsorption performance for all the activated carbons was obtained at pH 3.7, Mn-Carbon dose of 0.100g/100ml solution and contact time 4.5h. The adsorption isotherms fit the Langmuir isotherm equation. The kinetic data followed the pseudo-second-order model. The thermodynamic parameters indicated that the processes were spontaneous and endothermic. According to these results, the prepared Mn modified activated carbons are promising adsorbents for the removal of Neutral Red from wastewater.
Analysis of neutralizing epitopes on foot-and-mouth disease virus.
Pfaff, E; Thiel, H J; Beck, E; Strohmaier, K; Schaller, H
For the investigation of the antigenic determinant structure of foot-and-mouth disease virus (FMDV), neutralizing monoclonal antibodies (MAbs) against complete virus were characterized by Western blot (immunoblot), enzyme immunoassay, and competition experiments with a synthetic peptide, isolated coat protein VP1, and viral particles as antigens. Two of the four MAbs reacted with each of these antigens, while the other two MAbs recognized only complete viral particles and reacted only very poorly with the peptide. The four MAbs showed different neutralization patterns with a panel of 11 different FMDV strains. cDNA-derived VP1 protein sequences of the different strains were compared to find correlations between the primary structure of the protein and the ability of virus to be neutralized. Based on this analysis, it appears that the first two MAbs recognized overlapping sequential epitopes in the known antigenic site represented by the peptide, whereas the two other MAbs recognized conformational epitopes. These conclusions were supported and extended by structural analyses of FMDV mutants resistant to neutralization by an MAb specific for a conformational epitope. These results demonstrate that no amino acid exchanges had occurred in the primary antigenic site of VP1 but instead in the other coat proteins VP2 and VP3, which by themselves do not induce neutralizing antibodies. Images PMID:2835507
Neutralization activity of patient sera collected during the 2008-2009 Chikungunya outbreak in Thailand.
Kishishita, Natsuko; Sasayama, Mikiko; Takeda, Naokazu; Sa-Ngasang, Areerat; Anuegoonpipat, Atchareeya; Anantapreecha, Surapee
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.
Novel pyrazole derivatives as neutral CB₁ antagonists with significant activity towards food intake.
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
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.
Toward Effective HIV Vaccination INDUCTION OF BINARY EPITOPE REACTIVE ANTIBODIES WITH BROAD HIV NEUTRALIZING ACTIVITY
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
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.
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.
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
Occurrence of neutral endopeptidase activity in the cat carotid body and its significance in chemoreception.
Kumar, G K; Runold, M; Ghai, R D; Cherniack, N S; Prabhakar, N R
The carotid body contains both tachykinins and enkephalins. Neutral endopeptidase (NEP, E.C. 220.127.116.11), has been suggested to involve in the metabolism of these neuropeptides in several organs. In the present study we determined neutral endopeptidase activity of the cat carotid body and assessed its significance in chemoreception. The cytosolic and membrane fractions of the carotid body contained NEP-like activity whereas it occurred only in the membrane fractions of the superior cervical and the nodose ganglia. Phosphoramidon, thiorphan and metal ion chelators inhibited NEP-like activity of all the 3 tissues studied; other protease inhibitors, however, were ineffective. Close carotid body administration of phosphoramidon significantly potentiated the carotid body response to low PO2 but not to hypercapnia. The enhanced response to hypoxia following phosphoramidon was further augmented by naloxone, an enkephalin antagonist. These results demonstrate that the glomus tissue contains detectable amounts of NEP-like activity and its inhibition selectively affects the hypoxic response of the carotid body.
Modeling activities on the negative-ion-based Neutral Beam Injectors of the Large Helical Device
Agostinetti, P.; Antoni, V.; Chitarin, G.; Pilan, N.; Serianni, G.; Veltri, P.; Cavenago, M.; Nakano, H.; Takeiri, Y.; Tsumori, K.
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.
A single amino acid deletion in the matrix protein of porcine reproductive and respiratory syndrome virus confers resistance to a polyclonal swine antibody with broadly neutralizing activity.
Trible, Benjamin R; Popescu, Luca N; Monday, Nicholas; Calvert, Jay G; Rowland, Raymond R R
Assessment of virus neutralization (VN) activity in 176 pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV) identified one pig with broadly neutralizing activity. A Tyr-10 deletion in the matrix protein provided escape from broad neutralization without affecting homologous neutralizing activity. The role of the Tyr-10 deletion was confirmed through an infectious clone with a Tyr-10 deletion. The results demonstrate differences in the properties and specificities of VN responses elicited during PRRSV infection.
Recombinant Clostridium difficile toxin fragments as carrier protein for PSII surface polysaccharide preserve their neutralizing activity.
Romano, Maria R; Leuzzi, Rosanna; Cappelletti, Emilia; Tontini, Marta; Nilo, Alberto; Proietti, Daniela; Berti, Francesco; Costantino, Paolo; Adamo, Roberto; Scarselli, Maria
Clostridium difficile is a Gram-positive bacterium and is the most commonly diagnosed cause of hospital-associated and antimicrobial-associated diarrhea. Despite the emergence of epidemic C. difficile strains having led to an increase in the incidence of the disease, a vaccine against this pathogen is not currently available. C. difficile strains produce two main toxins (TcdA and TcdB) and express three highly complex cell-surface polysaccharides (PSI, PSII and PSIII). PSII is the more abundantly expressed by most C. difficile ribotypes offering the opportunity of the development of a carbohydrate-based vaccine. In this paper, we evaluate the efficacy, in naive mice model, of PSII glycoconjugates where recombinant toxins A and B fragments (TcdA_B2 and TcdB_GT respectively) have been used as carriers. Both glycoconjugates elicited IgG titers anti-PSII although only the TcdB_GT conjugate induced a response comparable to that obtained with CRM197. Moreover, TcdA_B2 and TcdB_GT conjugated to PSII retained the ability to elicit IgG with neutralizing activity against the respective toxins. These results are a crucial proof of concept for the development of glycoconjugate vaccines against C. difficile infection (CDI) that combine different C. difficile antigens to potentially prevent bacterial colonization of the gut and neutralize toxin activity.
Analysis of variola and vaccinia virus neutralization assays for smallpox vaccines.
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
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.
Polycistronic expression of human platelet factor 4 with heparin-neutralizing activity in Escherichia coli.
Duan, Yitao; Wang, Zhe; Wu, Wei; Fang, Zhenjiang; Huang, He
Human platelet factor 4 (hPF4) was evaluated as a clinical alternative to protamine for heparin neutralization, a protector against radiation injury and an anti-neoplastic. To achieve high-level expression of hPF4, expression vectors pET-28a(+)-nf PF4 (n=4, 5, 6) containing n tandem repeats of PF4 were constructed and transformed into the Escherichia coli BL21(DE3) strain. A higher expression level, about 45% of the total proteins (TP), was obtained for E. coli BL21(DE3)/pET28a(+)-nf PF4 (n=4, 5, 6). The purified His-PF4 protein was further identified by cleavage with enterokinase and MS, and its heparin-neutralizing activity was determined by colony formation assay. This study represents a novel approach to large-scale production of PF4 in E. coli, one that might be applied to large-scale production of PF4 protein for possible clinical application. It also provides theoretical points for the expression and purification of other small-molecule peptides.
Leake, James E.; Linton, Mark G.
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.
Brain Activity in Response to Trauma-specific, Negative, and Neutral Stimuli. A fMRI Study of Recent Road Traffic Accident Survivors
Nilsen, Andre S.; Blix, Ines; Leknes, Siri; Ekeberg, Øivind; Skogstad, Laila; Endestad, Tor; Østberg, Bjørn C.; Heir, Trond
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
Measurement of the total active 8B solar neutrino flux at the Sudbury Neutrino Observatory with enhanced neutral current sensitivity.
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
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.
Monte-Carlo Analysis of the Flavour Changing Neutral Current B \\to Gamma at Babar
Smith, D.
The main theme of this thesis is a Monte-Carlo analysis of the rare Flavour Changing Neutral Current (FCNC) decay b→sγ. The analysis develops techniques that could be applied to real data, to discriminate between signal and background events in order to make a measurement of the branching ratio of this rare decay using the BaBar detector. Also included in this thesis is a description of the BaBar detector and the work I have undertaken in the development of the electronic data acquisition system for the Electromagnetic calorimeter (EMC), a subsystem of the BaBar detector.
Lipolytic activity of ricin from Ricinus sanguineus and Ricinus communis on neutral lipids.
Lombard, S; Helmy, M E; Piéroni, G
The present study was carried out with a view of determining ricin lipolytic activity on neutral lipids in emulsion and in a membrane-like model. Using 2,3-dimercapto-1-propanol tributyrate (BAL-TC(4)) as substrate, the lipolytic activity of ricin was found to be proportional to ricin and substrate concentrations, with an apparent K(m) (K(m,app)) of 2.4 mM, a k(cat) of 200 min(-1) and a specific activity of 1.0 unit/mg of protein. This work was extended to p-nitrophenyl (pNP) fatty acid esters containing two to twelve carbon atoms. Maximum lipolytic activity was registered on pNP decanoate (pNPC(10)), with a K(m,app) of 3.5 mM, a k(cat) of 173 min(-1) and a specific activity of 3.5 units/mg of protein. Ricin lipolytic activity is pH and galactose dependent, with a maximum at pH 7.0 in the presence of 0.2 M galactose. Using the monolayer technique with dicaprin as substrate, ricin showed a lipolytic activity proportional to the ricin concentration at 20 mN/m, which is dependent on the surface pressure of the lipid monolayer and is detectable up to 30 mN/m, a surface pressure that is of the same order of magnitude as that of natural cell membranes. The methods based on pNPC(10) and BAL-TC(4) hydrolysis are simple and reproducible; thus they can be used for routine studies of ricin lipolytic activity. Ricin from Ricinus communis and R. sanguineus were treated with diethyl p-nitrophenylphosphate, an irreversible serine esterase inhibitor, and their lipolytic activities on BAL-TC(4) and pNPC(10), and cytotoxic activity, were concurrently recorded. A reduction in lipolytic activity was accompanied by a decrease in cytotoxicity on Caco2 cells. These data support the idea that the lipolytic activity associated with ricin is relevant to a lipase whose activity is pH and galactose dependent, sensitive to diethyl p-nitrophenylphosphate, and that a lipolytic step may be involved in the process of cell poisoning by ricin. Both colorimetric tests used in this study are sensitive
development of a neutral mass spectrometer dedicated to the analysis of planetary envelopes (NIMEIS)
NASA Astrophysics Data System (ADS)
Becker, J.
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.
Lipophilicity is a key factor to increase the antiviral activity of HIV neutralizing antibodies.
Augusto, Marcelo T; Hollmann, Axel; Troise, Fulvia; Veiga, Ana S; Pessi, Antonello; Santos, Nuno C
The HIV broadly neutralizing antibody 2F5 targets the transiently exposed epitope in the membrane proximal external region (MPER) of HIV-1 gp41, by a two-step mechanism involving the viral membrane and this viral glycoprotein. It was recently shown that 2F5 conjugation with a cholesterol moiety outside of the antibody paratope substantially increases its antiviral activity. Additionally, the antiviral activity of D5, a human antibody that binds to the N-terminal heptad repeat (NHR) of gp41 and lacks membrane binding, was boosted by the same cholesterol conjugation. In this work, we evaluated the membrane affinity of both antibodies towards membranes of different compositions, using surface plasmon resonance. A correlation was found between membrane affinity and antiviral activity against HIV-1. We propose that the conjugation of cholesterol to 2F5 or D5 allows a higher degree of antibody pre-concentration at the viral membrane. This way, the antibodies become more available to bind efficiently to the gp41 epitope, blocking viral fusion faster than the unconjugated antibody. These results set up a relevant strategy to improve the rational design of therapeutic antibodies against HIV.
Stabilisation analysis for switched neutral systems based on sampled-data control
NASA Astrophysics Data System (ADS)
Krishnasamy, R.; Balasubramaniam, P.
In this paper, the problem of stabilisation analysis for switched neutral systems based on sampled-data control and average dwell time approach is investigated. Delay-dependent stabilisation results are derived in terms of linear matrix inequalities by constructing piecewise Lyapunov-Krasovskii functional based on the Wirtinger's inequality. Also, the controller gain matrix is designed by applying an input-delay approach. Further convex combination technique and some integral inequalities are used to derive less conservative results. The effectiveness of the derived results is validated through numerical examples.
Optimization, characterization, sulfation and antitumor activity of neutral polysaccharides from the fruit of Borojoa sorbilis cuter.
Xu, Fangfang; Liao, Kangsheng; Wu, Yunshan; Pan, Qi; Wu, Lilan; Jiao, Hong; Guo, Dean; Li, Ben; Liu, Bo
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.
Performance of thermally activated dolomite for the treatment of Ni and Zn in contaminated neutral drainage.
Calugaru, Iuliana Laura; Neculita, Carmen Mihaela; Genty, Thomas; Bussière, Bruno; Potvin, Robin
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.
Light modulated activity of root alkaline/neutral invertase involves the interaction with 14-3-3 proteins.
Gao, Jing; van Kleeff, Paula J M; Oecking, Claudia; Li, Ka Wan; Erban, Alexander; Kopka, Joachim; Hincha, Dirk K; de Boer, Albertus H
Alkaline/neutral invertases (A/N-Invs) are now recognized as essential proteins in plant life. They catalyze the irreversible breakdown of sucrose into glucose and fructose and thus supply the cells with energy as well as signaling molecules. In this study we report on a mechanism that affects the activity of the cytosolic invertase AtCINV1 (At-A/N-InvG or AT1G35580). We demonstrate that Ser547 at the extreme C-terminus of the AtCINV1 protein is a substrate of calcium-dependent kinases (CPK3 and 21) and that phosphorylation creates a high-affinity binding site for 14-3-3 proteins. The invertase as such has basal activity, but we provide evidence that interaction with 14-3-3 proteins enhances its activity. The analysis of three quadruple 14-3-3 mutants generated from six T-DNA insertion mutants of the non-epsilon family shows both specificity as well as redundancy for this function of 14-3-3 proteins. The strong reduction in hexose levels in the roots of one 14-3-3 quadruple mutant plant is in line with the activating function of 14-3-3 proteins. The physiological relevance of this mechanism that affects A/N-invertase activity is underscored by the light-induced activation and is another example of the central role of 14-3-3 proteins in mediating dark/light signaling. The nature of the light-induced signal that travels from the shoot to root and the question whether this signal is transmitted via cytosolic Ca(++) changes that activate calcium-dependent kinases, await further study.
HIV-1 Dual Infected LTNP-EC Patients Developed an Unexpected Antibody Cross-Neutralizing Activity
Pernas, Maria; Sanchez-Merino, Victor; Casado, Concepcion; Merino-Mansilla, Alberto; Olivares, Isabel; Yuste, Eloisa; Lopez-Galindez, Cecilio
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
Novel neutralizing hedgehog antibody MEDI-5304 exhibits antitumor activity by inhibiting paracrine hedgehog signaling.
Michaud, Neil R; Wang, Youzhen; McEachern, Kristen A; Jordan, Jerold J; Mazzola, Anne Marie; Hernandez, Axel; Jalla, Sanjoo; Chesebrough, Jon W; Hynes, Mark J; Belmonte, Matthew A; Wang, Lidong; Kang, Jaspal S; Jovanovic, Jelena; Laing, Naomi; Jenkins, David W; Hurt, Elaine; Liang, Meina; Frantz, Christopher; Hollingsworth, Robert E; Simeone, Diane M; Blakey, David C; Bedian, Vahe
The hedgehog pathway has been implicated in the tumorigenesis, tumor progression, and metastasis of numerous human cancers. We generated the first fully human hedgehog antibody MEDI-5304 and characterized its antitumor activity and preclinical toxicology. MEDI-5304 bound sonic hedgehog (SHH) and Indian hedgehog (IHH) with low picomolar affinity and neutralized SHH and IHH activity in cellular mGLI1 reporter assays. The antibody inhibited transcription of hedgehog target genes and osteoblast differentiation of C3H10T1/2 cells. We evaluated the activity of MEDI-5304 in vivo in model systems that allowed us to evaluate two primary hypotheses of hedgehog function in human cancer, paracrine signaling between tumor and stromal cells and cancer stem cell (CSC) self-renewal. MEDI-5304 displayed robust pharmacodynamic effects in stromal cells that translated to antitumor efficacy as a single agent in an HT-29/MEF coimplantation model of paracrine hedgehog signaling. MEDI-5304 also improved responses to carboplatin in the HT-29/MEF model. The antibody, however, had no effect as a single agent or in combination with gemcitabine on the CSC frequency or growth of several primary pancreatic cancer explant models. These findings support the conclusion that hedgehog contributes to tumor biology via paracrine tumor-stromal signaling but not via CSC maintenance or propagation. Finally, the only safety study finding associated with MEDI-5304 was ondontodysplasia in rats. Thus, MEDI-5304 represents a potent dual hedgehog inhibitor suitable for continued development to evaluate efficacy and safety in human patients with tumors harboring elevated levels of SHH or IHH.
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.
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.
Generation of antitumor active neutral medium-sized alpha-glycan in apple vinegar fermentation.
Abe, Kaoru; Kushibiki, Toshisada; Matsue, Hajime; Furukawa, Ken-Ichi; Motomura, Shigeru
The physiologically active substances in apple vinegar have not yet been chemically characterized. We studied the biological functions of apple vinegar produced from crushed apples, and found that the constituent neutral medium-sized alpha-glycan (NMalphaG) acts as an antitumor agent against experimental mouse tumors. NMalphaG is a homoglycan composed of glucose having a molecular weight of about 10,000 and a branched structure bearing alpha (1-4,6) linkages. In this study, we clarified the origin of NMalphaG in apple vinegar by examination of its content in alcohol and acetic acid fermentation products sequentially. We found that NMalphaG appeared in acetic acid fermentation, but not in alcohol fermentation. Furthermore we investigated NMalphaG origin using acetic acid fermentation from alcohol fortifiied apple without alcohol fermentation and from raw material with varying amounts of pomace. The results indicate that NMalphaG originated in the apple fruit body and that its production requires both fermentation processes.
Opportunities to exploit non-neutralizing HIV-specific antibody activity
Ackerman, Margaret E.; Alter, Galit
Antibodies act as a nexus between innate and adaptive immunity: they provide a means to engage a spectrum of innate immune effector cells in order to clear viral particles and infected cells, and prime antigen presentation. This functional landscape is remarkably complex, and depends on antibody isotype, subclass, and glycosylation; the expression levels and patterns of a suite of Fc receptors with both complementary and opposing activities; and a host of innate immune cells capable of differential responses to opsonized particles and present at different sites. In vivo, even neutralizing antibodies rely on their ability to act as molecular beacons and recruit innate immune effector cells in order to provide protection, and results from both human and macaque studies have implicated these effector functions in vaccine-mediated protection. Thus, while enhancing effector function is a tractable handle for potentiating antibody-mediated protection from HIV infection, success will depend critically on leveraging understanding of the means by which antibodies with specific functional profiles could be elicited, which effector functions could provide optimal protection, and perhaps most critically, how to efficiently recruit the innate effector cells present at sites of infection. PMID:24191934
Mean square delay dependent-probability-distribution stability analysis of neutral type stochastic neural networks.
Muralisankar, S; Manivannan, A; Balasubramaniam, P
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.
Evolution of broadly cross-reactive HIV-1-neutralizing activity: therapy-associated decline, positive association with detectable viremia, and partial restoration of B-cell subpopulations.
Ferreira, Carolina B; Merino-Mansilla, Alberto; Llano, Anuska; Pérez, Ignacio; Crespo, Isabel; Llinas, Laia; Garcia, Felipe; Gatell, Jose M; Yuste, Eloisa; Sanchez-Merino, Victor
Little is known about the stability of HIV-1 cross-neutralizing responses. Taking into account the fact that neutralization breadth has been positively associated with plasma viral load, there is no explanation for the presence of broadly neutralizing responses in a group of patients on treatment with undetectable viremia. In addition, the B-cell profile responsible for broadly cross-neutralizing responses is unknown. Here we studied the evolution of neutralizing responses and the B-cell subpopulation distribution in a group of patients with broadly cross-reactive HIV-1-neutralizing activity. We studied neutralization breadth evolution in a group of six previously identified broadly cross-neutralizing patients and six control patients during a 6-year period with a previously described minipanel of recombinant viruses from five different subtypes. B-cell subpopulation distribution during the study was also determined by multiparametric flow cytometry. Broadly cross-neutralizing activity was transient in four broad cross-neutralizers and stable, up to 4.6 years, in the other two. In four out of five broad cross-neutralizers who initiated treatment, a neutralization breadth loss occurred after viremia had been suppressed for as much as 20 months. B-cell subpopulation analyses revealed a significant increase in the frequency of naive B cells in broadly cross-reactive samples, compared with samples with less neutralization breadth (increased from 44% to 62%). We also observed a significant decrease in tissue-like and activated memory B cells (decreased from 19% to 12% and from 17% to 9%, respectively). Our data suggest that HIV-1 broadly cross-neutralizing activity is variable over time and associated with detectable viremia and partial B-cell restoration.
Endotoxin- and ATP-neutralizing activity of alkaline phosphatase as a strategy to limit neuroinflammation
Background Alkaline phosphatase (AP) is a ubiquitously expressed enzyme which can neutralize endotoxin as well as adenosine triphosphate (ATP), an endogenous danger signal released during brain injury. In this study we assessed a potential therapeutic role for AP in inhibiting neuroinflammation using three complementary approaches. Methods Mice were immunized to induce experimental autoimmune encephalomyelitis (EAE) and treated with AP for seven days during different phases of disease. In addition, serological assays to determine AP activity, endotoxin levels and endotoxin-reactive antibodies were performed in a cohort of multiple sclerosis (MS) patients and controls. Finally, the expression of AP and related enzymes CD39 and CD73 was investigated in brain tissue from MS patients and control subjects. Results AP administration during the priming phase, but not during later stages, of EAE significantly reduced neurological signs. This was accompanied by reduced proliferation of splenocytes to the immunogen, myelin oligodendrocyte glycoprotein peptide. In MS patients, AP activity and isoenzyme distribution were similar to controls. Although endotoxin-reactive IgM was reduced in primary-progressive MS patients, plasma endotoxin levels were not different between groups. Finally, unlike AP and CD73, CD39 was highly upregulated on microglia in white matter lesions of patients with MS. Conclusions Our findings demonstrate that: 1) pre-symptomatic AP treatment reduces neurological signs of EAE; 2) MS patients do not have altered circulating levels of AP or endotoxin; and 3) the expression of the AP-like enzyme CD39 is increased on microglia in white matter lesions of MS patients. PMID:23231745
Relationships between adaptive and neutral genetic diversity and ecological structure and functioning: a meta-analysis
Whitlock, Raj
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
Non-Cationic Proteins Are Associated with HIV Neutralizing Activity in Genital Secretions of Female Sex Workers
Birse, Kenzie D. M.; Cole, Amy L.; Hirbod, Taha; McKinnon, Lyle; Ball, Terry B.; Westmacott, Garrett R.; Kimani, Joshua; Plummer, Frank; Cole, Alexander M.; Burgener, Adam; Broliden, Kristina
Objective Cationic proteins found in cervicovaginal secretions (CVS) are known to contribute to the early antiviral immune response against HIV-infection in vitro. We here aimed to define additional antiviral factors that are over-expressed in CVS from female sex workers at high risk of infection. Methods CVS were collected from Kenyan HIV-seronegative (n = 34) and HIV-seropositive (n = 12) female sex workers, and were compared with those from HIV-seronegative low-risk women (n = 12). The highly exposed seronegative (HESN) sex workers were further divided into those with less (n = 22) or more (n = 12) than three years of documented sex work. Cationic protein-depleted CVS were assessed for HIV-neutralizing activity by a PBMC-based HIV-neutralizing assay, and then characterized by proteomics. Results HIV neutralizing activity was detected in all unprocessed CVS, however only CVS from the female sex worker groups maintained its HIV neutralizing activity after cationic protein-depletion. Differentially abundant proteins were identified in the cationic protein-depleted secretions including 26, 42, and 11 in the HESN>3yr, HESN<3yr, and HIV-positive groups, respectively. Gene ontology placed these proteins into functional categories including proteolysis, oxidation-reduction, and epidermal development. The proteins identified in this study include proteins previously associated with the HESN phenotype in other cohorts as well as novel proteins not yet associated with anti-HIV activities. Conclusion While cationic proteins appear to contribute to the majority of the intrinsic HIV neutralizing activity in the CVS of low-risk women, a broader range of non-cationic proteins were associated with HIV neutralizing activity in HESN and HIV-positive female sex workers. These results indicate that novel protein factors found in CVS of women with high-risk sexual practices may have inherent antiviral activity, or are involved in other aspects of anti-HIV host defense, and warrant
Neutralization Assay for Chikungunya Virus Infection: Plaque Reduction Neutralization Test.
Azami, Nor Azila Muhammad; Moi, Meng Ling; Takasaki, Tomohiko
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.
Smith, T
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
Strategic addition of an N-linked glycan to a monoclonal antibody improves its HIV-1-neutralizing activity.
Song, Ruijiang; Oren, Deena A; Franco, David; Seaman, Michael S; Ho, David D
Ibalizumab is a humanized monoclonal antibody that binds human CD4--a key receptor for HIV--and blocks HIV-1 infection. However, HIV-1 strains with mutations resulting in loss of an N-linked glycan from the V5 loop of the envelope glycoprotein gp120 are resistant to ibalizumab. Previous structural analysis suggests that this glycan fills a void between the gp120 V5 loop and the ibalizumab light chain, perhaps causing steric hindrance that disrupts viral entry. If this void contributes to HIV-1 resistance to ibalizumab, we reasoned that 'refilling' it by engineering an N-linked glycan into the ibalizumab light chain at a position spatially proximal to gp120 V5 may restore susceptibility to ibalizumab. Indeed, one such ibalizumab variant neutralized 100% of 118 diverse HIV-1 strains tested in vitro, including 10 strains resistant to parental ibalizumab. These findings demonstrate that the strategic placement of a glycan in the variable region of a monoclonal antibody can substantially enhance its activity.
Strategic addition of an N-linked glycan to a monoclonal antibody improves its HIV-1-neutralizing activity
Song, Ruijiang; Oren, Deena A.; Franco, David; Seaman, Michael S.; Ho, David D.
Ibalizumab is a humanized monoclonal antibody that binds human CD4—a key receptor for HIV—and blocks HIV-1 infection. However, HIV-1 strains with mutations resulting in loss of an N-linked glycan from the V5 loop of the envelope protein gp120 are resistant to ibalizumab. Previous structural analysis suggests that this glycan fills a void between the gp120 V5 loop and the ibalizumab L chain, perhaps causing steric hindrance that disrupts viral entry. If this void contributes to HIV-1 resistance to ibalizumab, we reasoned that 'refilling' it by engineering an N-linked glycan into the ibalizumab L chain at a position spatially proximal to gp120 V5 may restore susceptibility to ibalizumab. Indeed, one such ibalizumab variant neutralized 100% of 118 tested diverse HIV-1 strains in vitro, including ten strains resistant to parental ibalizumab. These findings demonstrate that the strategic placement of a glycan in the variable region of a monoclonal antibody can substantially enhance its activity. PMID:24097413
A role for calcium in the regulation of neutral trehalase activity in the fission yeast Schizosaccharomyces pombe.
Franco, Alejandro; Soto, Teresa; Vicente-Soler, Jero; Paredes, Vanessa; Madrid, Marisa; Gacto, Mariano; Cansado, José
Neutral trehalases mobilize trehalose accumulated by fungal cells as a protective and storage carbohydrate. A structural feature of these enzymes is the presence of an EF-like motif similar to that shown by many Ca2+-binding proteins. In this study we provide direct evidence for physical binding of Ca2+ to neutral trehalase (Ntp1p) of the fission yeast Schizosaccharomyces pombe, and show that aspartic residues at positions 97 and 108 in the conserved putative Ca2+-binding motif of Ntp1p appear to be responsible for this interaction. Mutations in these residues do not interfere with the ability of Ntp1p to associate in vivo with trehalose-6-phosphate synthase, but prevent activation of neutral trehalase triggered by the addition of glucose or by subjecting cells to stressing conditions. Strains expressing Ntp1p variants that are unable to bind Ca2+ partially resemble those devoid of the ntp1+ gene in terms of trehalose hyperaccumulation. Gel filtration of cell extracts from wild-type cells after EDTA treatment or from cells containing Ntp1p with mutations in aspartic acid residues within the Ca2+-binding site revealed that Ntp1p eluted mainly in an inactive conformation instead of the dimeric or trimeric active form of the enzyme. These results suggest that activation of S. pombe Ntp1p under different conditions depends upon Ca2+ binding through the Ca2+-binding motif as a prerequisite for correct enzyme oligomerization to its active form. Given the high degree of conservation of the Ca2+ accommodation site, this might be a general mechanism regulating neutral trehalase activity in other yeasts and filamentous fungi. PMID:12943532
Effects of Hypothermia and Self-Warming on Activity of Ca(2+)-Dependent Neutral Proteases in Tissues of Ground Squirrels and Rats.
Nurmagomedova, P M; Emirbekov, E Z; Abasova, M M
Effects of hypothermia and subsequent self-warming on activity of Ca(2+)-dependent neutral proteases were studied in tissues of ground squirrels and rats. Moderate hypothermia did not significantly change activity of Ca(2+)-dependent neutral proteases in the analyzed tissues of ground squirrels, but reduced protease activity in rat heart. Severe hypothermia reduced enzyme activity in the analyzed tissues of rats and ground squirrels. Differences in activity of Ca(2+)-dependent neutral proteases after long-term hypothermia and subsequent self-warming were found only in the heart.
Engineering HIV envelope protein to activate germline B cell receptors of broadly neutralizing anti-CD4 binding site antibodies.
McGuire, Andrew T; Hoot, Sam; Dreyer, Anita M; Lippy, Adriana; Stuart, Andrew; Cohen, Kristen W; Jardine, Joseph; Menis, Sergey; Scheid, Johannes F; West, Anthony P; Schief, William R; Stamatatos, Leonidas
Broadly neutralizing antibodies (bnAbs) against HIV are believed to be a critical component of the protective responses elicited by an effective HIV vaccine. Neutralizing antibodies against the evolutionarily conserved CD4-binding site (CD4-BS) on the HIV envelope glycoprotein (Env) are capable of inhibiting infection of diverse HIV strains, and have been isolated from HIV-infected individuals. Despite the presence of anti-CD4-BS broadly neutralizing antibody (bnAb) epitopes on recombinant Env, Env immunization has so far failed to elicit such antibodies. Here, we show that Env immunogens fail to engage the germline-reverted forms of known bnAbs that target the CD4-BS. However, we found that the elimination of a conserved glycosylation site located in Loop D and two glycosylation sites located in variable region 5 of Env allows Env-binding to, and activation of, B cells expressing the germline-reverted BCRs of two potent broadly neutralizing antibodies, VRC01 and NIH45-46. Our results offer a possible explanation as to why Env immunogens have been ineffective in stimulating the production of such bNAbs. Importantly, they provide key information as to how such immunogens can be engineered to initiate the process of antibody-affinity maturation against one of the most conserved Env regions.
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.
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.
Disproportionation and nuclease activity of bis[2-ethyl-2-hydroxybutanoato(2-)]oxochromate(V) in neutral aqueous solutions.
Levina, A; Lay, P A; Dixon, N E
Complex 1, [Cr(V)O(ehba)2]- (ehba = 2-ethyl-2-hydroxybutanoate(2-)) is the most studied model compound of relevance to the biological activity of Cr(V) with regard to Cr-induced cancers. The first detailed kinetic study of disproportionation of 1 under neutral pH conditions (pH 6.0-8.0, [NaClO4] = 1.0 M, 37 degrees C) is reported. Kinetic data were collected by stopped-flow and conventional UV-vis spectroscopies and processed by the global analysis method. The disproportionation, which follows the stoichiometry 3Cr(V) --> 2Cr(VI) + Cr(III) (1), leads to release of 5 mol of H+/3 mol of Cr(V). Reaction 1 is accelerated by phosphate, but is not affected by acetate, HEPES, or Tris buffers. Initial rates of Cr(V) decay are directly proportional to [Cr(V)]0 (0.020-1.0 mM); they increase with an increase in the pH values and decrease in the presence of a large excess of ehba ligand. The first direct evidence for the formation of Cr(IV) intermediates in reaction 1 has been obtained; however, their UV-vis spectral properties were different from those of the well-characterized Cr(IV)-ehba complexes. The Cr(III) products of reaction I in phosphate buffers differ from those in the other buffers. A mechanism is proposed for reaction 1 on the basis of kinetic modeling. Influences of the reaction time and conditions on the extent of plasmid DNA cleavage induced by 1 have been studied under conditions corresponding to those of the kinetic studies. A comparison of the kinetic and DNA cleavage results has shown that direct interaction of 1 with the phosphate backbone of DNA is the most likely first step in the mechanism of DNA cleavage in neutral media. Small additions of Mn(II) ((0.01-0.1)[Cr(V)]0) did not affect the rate and stoichiometry of reaction 1, but suppressed the formation of Cr(IV) intermediates (presumably due to the catalysis of Cr(IV) disproportionation). However, much higher concentrations of Mn(II) ((0.1-1.0)[Cr(V)]0) were required to inhibit DNA cleavage induced by
Characterization of a neutral and thermostable glucoamylase from the thermophilic mold Thermomucor indicae-seudaticae: activity, stability, and structural correlation.
Kumar, Pardeep; Islam, Asimul; Ahmad, Faizan; Satyanarayana, T
Glucoamylase from the thermophilic mold Thermomucor indicae-seudaticae was purified by anion exchange and gel filtration chromatographic techniques using a fast protein liquid chromatographic system. The structure and thermal stability of this unique 'thermostable and neutral glucoamylase' were analyzed by circular dichroism (CD). T. indicae-seudaticae glucoamylase (TGA) contained typical aromatic amino acid (tryptophan/tyrosine) fingerprints in its tertiary structure. Analysis of the far-UV CD spectrum at pH 7.0 and 25 degrees Celsius revealed the presence of 45% alpha-helix, 43% beta-sheet, and 12% remaining structures. The alpha-helix content was highest at pH 7.0, where glucoamylase is optimally active. This observation points towards the possible (alpha/alpha)(6) barrel catalytic domain in TGA, as reported in microbial glucoamylases. Thermal denaturation curves of the pure protein at different pH values revealed maximum stability at pH 7.0, where no change in the secondary structure was observed upon heating in the temperature range between 20 degrees Celsius and 60 degrees Celsius. The observed midpoint of thermal denaturation (T (m)) of glucoamylase at pH 7.0 was 67.1 degrees Celsius, which decreased on either sides of this pH. Thermostability of TGA enhanced in the presence of starch (0.1%) as no transition curve was obtained in the temperature range between 20 degrees Celsius and 85 degrees Celsius. The only product of TGA action on starch was glucose, and it did not exhibit transglycosylation activity even at 40% glucose that can also be considered as an advantage during starch saccharification.
Analysis of Potential Glove-Induced Hand Injury Metrics During Typical Neutral Buoyancy Training Operations
NASA Technical Reports Server (NTRS)
McFarland, Shane
Injuries to the hands are common among astronauts who train for extravehicular activity. When the gloves are pressurized, they restrict movement and create pressure points during tasks, sometimes resulting in pain, muscle fatigue, abrasions, and occasionally more severe injuries such as onycholysis. A brief review of NASA's Lifetime Surveillance of Astronaut Health's injury database reveals that 76 percent of astronaut hand and arm injuries from training between 1993 and 2010 occurred either to the fingernail, finger crotch, metacarpophalangeal joint, or fingertip. The purpose of this study was to assess the potential of using small sensors to measure forces acting on the fingers and hand within pressurized gloves and other variables such as skin temperature, humidity, and fingernail strain of a NASA crewmember during typical NBL (Neutral Buoyancy Laboratory) training activity. During the 5-hour exercise, the crewmember seemed to exhibit very large forces on some fingers, resulting in higher strain than seen in previous glove-box testing. In addition, vital information was collected on the glove cavity environment with respect to temperature and humidity. All of this information gathered during testing will be carried forward into future testing, potentially in glove-box- or 1G- (1 gravitational force) or NBL-suited environments, to better characterize and understand the possible causes of hand injury amongst NASA crew.
Carboxylic acid functional group analysis using constant neutral loss scanning-mass spectrometry.
Dron, Julien; Eyglunent, Gregory; Temime-Roussel, Brice; Marchand, Nicolas; Wortham, Henri
The present study describes the development of a new analytical technique for the functional group determination of the carboxylic moiety using atmospheric pressure chemical ionization-mass spectrometry (APCI-MS/MS) operated in the constant neutral loss scanning (CNLS) mode. Carboxylic groups were first derivatized into their corresponding methyl esters by reacting with BF3/methanol mix and the reaction mixture was then directly injected into the APCI chamber. The loss of methanol (m/z = 32 amu) resulting from the fragmentation of the protonated methyl esters was then monitored. Applying this method together with a statistical approach to reference mixtures containing 31 different carboxylic acids at randomly calculated concentrations demonstrated its suitability for quantitative functional group measurements with relative standard deviations below 15% and a detection limit of 0.005 mmol L(-1). Its applicability to environmental matrices was also shown through the determination of carboxylic acid concentrations inside atmospheric aerosol samples. To the best of our knowledge, it is the first time that the tandem mass spectrometry was successfully applied to functional group analysis, offering great perspectives in the characterization of complex mixtures which are prevailing in the field of environmental analysis as well as in the understanding of the chemical processes occurring in these matrices.
Organic matter in sediment layers of an acidic mining lake as assessed by lipid analysis. Part II: Neutral lipids.
Poerschmann, Juergen; Koschorreck, Matthias; Górecki, Tadeusz
Natural neutralization of acidic mining lakes is often limited by organic matter. The knowledge of the sources and degradability of organic matter is crucial for understanding alkalinity generation in these lakes. Sediments collected at different depths (surface sediment layer from 0 to 1 cm and deep sediment layer from 4 to 5cm) from an acidic mining lake were studied in order to characterize sedimentary organic matter based on neutral signature markers. Samples were exhaustively extracted, subjected to pre-chromatographic derivatizations and analyzed by GC/MS. Herein, molecular distributions of diagnostic alkanes/alkenes, terpenes/terpenoids, polycyclic aromatic hydrocarbons, aliphatic alcohols and ketones, sterols, and hopanes/hopanoids were addressed. Characterization of the contribution of natural vs. anthropogenic sources to the sedimentary organic matter in these extreme environments was then possible based on these distributions. With the exception of polycyclic aromatic hydrocarbons, combined concentrations across all marker classes proved higher in the surface sediment layer as compared to those in the deep sediment layer. Alkane and aliphatic alcohol distributions pointed to predominantly allochthonous over autochthonous contribution to sedimentary organic matter. Sterol patterns were dominated by phytosterols of terrestrial plants including stigmasterol and β-sitosterol. Hopanoid markers with the ββ-biohopanoid "biological" configuration were more abundant in the surface sediment layer, which pointed to higher bacterial activity. The pattern of polycyclic aromatic hydrocarbons pointed to prevailing anthropogenic input. Pyrolytic makers were likely to due to atmospheric deposition from a nearby former coal combustion facility. The combined analysis of the array of biomarkers provided new insights into the sources and transformations of organic matter in lake sediments.
Immobilized carbon nanotubes as matrix for MALDI-TOF-MS analysis: applications to neutral small carbohydrates.
Ren, Shi-fang; Zhang, Li; Cheng, Zhi-hong; Guo, Yin-long
In this work, we reported on the advantages of immobilized carbon nanotubes as a novel MALDI-matrix. Recently, carbon nanotubes have been reported to be an effective MALDI matrix for small molecules (Anal. Chem.2003, 75, 6191), as it can eliminate the interfering matrix peaks as well as form a web morphology to fully disperse the analyte and allow strong ultraviolet absorption for enhanced pulsed laser desorption and ionization. In our study, to overcome the problem that the carbon nanotube matrix may fly off from the target, a type of polyurethane adhesive, NIPPOLAN-DC-205, is introduced to immobilize carbon nanotubes on the target, which enables widespread application of carbon nanotubes as matrix for MALDI-MS analysis. At the same time, the properties of the carbon nanotubes as an efficient matrix remained after immobilization. The presence of NIPPOLAN-DC-205 increases the time for analysis at a particular desorption spot by minimizing the time-consuming search for "hot spots" and facilitating experiments such as post source decay (PSD) which need longer-lasting signals. Moreover, NIPPOLAN-DC-205 produces no interference peaks and can easily be cleaned with acetone. Fast evaporation technology may be used to enhance signal reproducibility in MALDI analysis using carbon nanotubes as matrix. Consequently, the applicability of the carbon nanotube as matrix for matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis of low molecular mass analytes is highly improved. The feasibility of the method employing polyurethane is demonstrated by comparison of the results produced from the carbon nanotube matrix with and without immobilization. In addition, neutral small carbohydrates, which are difficult to be ionized normally, can be cationized with high efficiency by MALDI-TOF-MS using the immobilized carbon nanotube matrix. The method was further applied to analyze peptides and detect urine glucose successfully.
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.
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
Morin Attenuates Streptococcus suis Pathogenicity in Mice by Neutralizing Suilysin Activity
Li, Gen; Lu, Gejin; Qi, Zhimin; Li, Hongen; Wang, Lin; Wang, Yanhui; Liu, Bowen; Niu, Xiaodi; Deng, Xuming; Wang, Jianfeng
Streptococcus suis, a Gram-positive pathogen, is widely recognized as an important agent of swine infection, and it is also known to cause a variety of zoonoses, such as meningitis, polyarthritis and pneumonia. Suilysin (SLY), an extracellular pore-forming toxin that belongs to the cholesterol-dependent cytolysin family, is an essential virulence factor of S. suis capsular type 2 (SS2). Here, we found that morin hydrate (morin), a natural flavonoid that lacks anti-SS2 activity, inhibits the hemolytic activity of SLY, protects J774 cells from SS2-induced injury and protects mice from SS2 infection. Further, by molecular modeling and mutational analysis, we found that morin binds to the "stem" domain 2 in SLY and hinders its transformation from the monomer form to the oligomer form, which causes the loss of SLY activity. Our study demonstrates that morin hinders the cell lysis activity of SLY through a novel mechanism of interrupting the heptamer formation. These findings may lead to the development of promising therapeutic candidates for the treatment of SS2 infections. PMID:28373868
Efficient neutralizing activity of cocktailed recombinant human antibodies against hepatitis A virus infection in vitro and in vivo.
Cao, Jingyuan; Meng, Shufang; Li, Chuan; Ji, Yan; Meng, Qingling; Zhang, Quanfu; Liu, Feng; Li, Jiandong; Bi, Shengli; Li, Dexin; Liang, Mifang
Hepatitis A virus (HAV) is the major pathogen responsible for acute infectious hepatitis A, a disease that is prevalent worldwide. Although HAV immunization effectively prevents infection, primary immunizations must be administered at least 2 weeks prior to HAV exposure. In contrast, passive immunization with pooled human immunoglobulin (Ig) can provide immediate and rapid protection from HAV infection. Because the use of human sera-derived Igs carries the risk of contamination, we sought to develop recombinant HAV-neutralizing human antibodies. We prepared a combinatorial phage display library of recombinant human anti-HAV antibodies from RNA extracted from the blood lymphocytes of a convalescent hepatitis A patient. Two recombinant human IgG antibodies, HAIgG16 and HAIgG78, were screened from the antibody library by their ability to bind with high affinity to purified, inactivated HAV virions. These antibodies recognized different epitopes of the HAV virion capsid, and competed with both patient sera and well-characterized neutralizing mouse monoclonal antibodies. A cocktailed mixture of HAIgG16 and HAIgG78 at a 3:1 ratio was prepared to compare its combined biological activity with that conferred by each antibody individually. The cocktailed antibodies displayed a stronger neutralizing activity in vitro than that observed with either HAIgG16 and HAIgG78 alone. To determine the in vivo neutralizing abilities of these antibodies, rhesus monkeys were inoculated with cocktailed antibodies and challenged with HAV. Whereas control animals developed hepatitis A and seroconverted to the HAV antibody, animals receiving cocktailed antibodies were protected either from viral infection or from developing clinical hepatitis. These results demonstrate that recombinant human antibody preparations could be used to prevent or treat early-stage HAV infection.
Analysis of Power Distribution on Beamline Components at Different Neutralization Efficiencies on NBI Test Stand
NASA Astrophysics Data System (ADS)
Li, Xiang; Xu, Yongjian; Yu, Ling; Chen, Yu; Hu, Chundong; Tao, Ling
Neutral beam injection is recognized as one of the most effective means for plasma heating. According to the research plan of the EAST physics experiment, two sets of neutral beam injector (4-8 MW, 10-100 s) were built and operated in 2014. Neutralization efficiency is one of the important parameters for neutral beam. High neutralization efficiency can not only improve injection power at the same beam energy, but also decrease the power deposited on the heat-load components in the neutral beam injector (NBI). This research explores the power deposition distribution at different neutralization efficiencies on the beamline components of the NBI device. This work has great significance for guiding the operation of EAST-NBI, especially in long pulse and high power operation, which can reduce the risk of thermal damage of the beamline components and extend the working life of the NBI device. supported by the International Science and Technology Cooperation Program of China (No. 2014DFG61950), National Natural Science Foundation of China (No. 11405207) and the Foundation of ASIPP (No. DSJJ-15-GC03)
Meta-analysis of neutralizing antibody conversion with onabotulinumtoxinA (BOTOX®) across multiple indications.
Naumann, Markus; Carruthers, Alastair; Carruthers, Jean; Aurora, Sheena K; Zafonte, Ross; Abu-Shakra, Susan; Boodhoo, Terry; Miller-Messana, Mary Ann; Demos, George; James, Lynn; Beddingfield, Frederick; VanDenburgh, Amanda; Chapman, Mary Ann; Brin, Mitchell F
This meta-analysis evaluated the frequency of neutralizing antibody (nAb) conversion with onabotulinumtoxinA (BOTOX®; Allergan) across five studied indications. The analysis was based on large, controlled or prospective, open-label trials (durations 4 months to ≥2 years). Serum samples were analyzed for nAbs using the Mouse Protection Assay. Subjects who were antibody negative at baseline and had at least one analyzable postbaseline antibody assay result were included. The 16 clinical studies included 3,006 subjects; of these, 2,240 met the inclusion criteria for this analysis. Subjects received 1-15 treatments (mean 3.8 treatments) with onabotulinumtoxinA. Total doses per treatment cycle ranged from 10 or 20 units in glabellar lines to 20-500 units in cervical dystonia. The numbers of subjects who converted from an antibody-negative status at baseline to antibody-positive status at any post-treatment time point were: cervical dystonia 4/312 (1.28%), glabellar lines 2/718 (0.28%), overactive bladder 0/22 (0%), post-stroke spasticity 1/317 (0.32%), and primary axillary hyperhidrosis 4/871 (0.46%). Across all indications, 11/2,240 subjects (0.49%) converted from antibody negative at baseline to positive at one or more post-treatment time points, but only three subjects became clinically unresponsive to onabotulinumtoxinA at some point following a positive assay. Based on these large trials, the frequency of antibody conversion after onabotulinumtoxinA treatment is very low, and infrequently leads to loss of efficacy. © 2010 Movement Disorder Society.
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
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.
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
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.
Differential neutralizing activities of a single domain camelid antibody (VHH) specific for ricin toxin's binding subunit (RTB).
Herrera, Cristina; Vance, David J; Eisele, Leslie E; Shoemaker, Charles B; Mantis, Nicholas J
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.
Influences of acidic reaction and hydrolytic conditions on monosaccharide composition analysis of acidic, neutral and basic polysaccharides.
Wang, Qing-Chi; Zhao, Xia; Pu, Jiang-Hua; Luan, Xiao-Hong
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.
Canine distemper virus neutralization activity is low in human serum and it is sensitive to an amino acid substitution in the hemagglutinin protein
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.
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.
Design of multi-megawatt actively cooled beam dumps for the Neutral-Beam Engineering Test Facility
Paterson, J.A.; Koehler, G.; Wells, R.P.
The Neutral Beam Engineering Test Facility will test Neutral Beam Sources up to 170 keV, 65 Amps, with 30 second beam-on times. For this application actively cooled beam dumps for both the neutral and ionized particles will be required. The dumps will be able to dissipate a wide range of power density profiles by utilizing a standard modular panel design which is incorporated into a moveable support structure. The thermal hydraulic design of the panels permit the dissipation of 2 kW/cm/sup 2/ anywhere on the panel surface. The water requirements of the dumps are optimized by restricting the flow to panel sections where the heat flux falls short of the design value. The mechanical design of the beam-dump structures is described along with tests performed on a prototype panel. The prototype tests were performed on two different panel designs, one manufactured by Mc Donnell Douglas (MDAC) the other by United Technologies (UT). The dissipation capabilities of the panels were tested at the critical regions to verify their use in the beam dump assemblies.
Antimicrobial activity of neutralized extracellular culture filtrates of lactic acid bacteria isolated from a cultured Indian milk product ('dahi').
Varadaraj, M C; Devi, N; Keshava, N; Manjrekar, S P
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.
Capillary gas chromatographic analysis of alditol acetates of neutral and amino sugars in bacterial cell walls.
Fox, A; Morgan, S L; Hudson, J R; Zhu, Z T; Lau, P Y
Several improvements in the preparation of alditol acetates of neutral and amino sugars and in the preparation of glass capillary columns for their separation are described. Modifications in sample preparation permitted the simultaneous processing of multiple samples and eliminated extraneous background peaks. Efficient and inert columns were tailor-made for the separation of alditol acetates of neutral and amino sugars by leaching glass capillaries with aqueous hydrochloric acid and dynamically coating with SP-2330.
Effects of acute psychosocial stress on neural activity to emotional and neutral faces in a face recognition memory paradigm.
Li, Shijia; Weerda, Riklef; Milde, Christopher; Wolf, Oliver T; Thiel, Christiane M
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.
NLTE Analysis of High-resolution H-band Spectra. II. Neutral Magnesium
NASA Astrophysics Data System (ADS)
Zhang, Junbo; Shi, Jianrong; Pan, Kaike; Allende Prieto, Carlos; Liu, Chao
Aiming at testing the validity of our magnesium atomic model and investigating the effects of non-local thermodynamical equilibrium (NLTE) on the formation of the H-band neutral magnesium lines, we derive the differential Mg abundances from selected transitions for 13 stars either adopting or relaxing the assumption of local thermodynamical equilibrium (LTE). Our analysis is based on high-resolution and high signal-to-noise ratio H-band spectra from the Apache Point Observatory Galactic Evolution Experiment (APOGEE) and optical spectra from several instruments. The absolute differences between the Mg abundances derived from the two wavelength bands are always less than 0.1 dex in the NLTE analysis, while they are slightly larger for the LTE case. This suggests that our Mg atomic model is appropriate for investigating the NLTE formation of the H-band Mg lines. The NLTE corrections for the Mg i H-band lines are sensitive to the surface gravity, becoming larger for smaller log g values, and strong lines are more susceptible to departures from LTE. For cool giants, NLTE corrections tend to be negative, and for the strong line at 15765 Å they reach ‑0.14 dex in our sample, and up to ‑0.22 dex for other APOGEE stars. Our results suggest that it is important to include NLTE corrections in determining Mg abundances from the H-band Mg i transitions, especially when strong lines are used. Based on observations collected on the 2.16 m telescope at Xinglong station, National Astronomical Observatories, Chinese Academy of Sciences, the 2.2 m telescope at the Calar Alto Observatory, the 1.88 m reflector on the Okayama Astrophysical Observatory, the Kitt Peak coudé feed telescope, and the McMath–Pierce solar telescope and the coudé focus of the Mayall 4 m reflector at Kitt Peak.
Application of subtracted gDNA microarray-assisted Bulked Segregant Analysis for rapid discovery of molecular markers associated with day-neutrality in strawberry (Fragaria x ananassa)
Gor, Mian Chee; Mantri, Nitin; Pang, Edwin
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
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
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.
Fishbone activity in experimental advanced superconducting tokamak neutral beam injection plasma
Xu, Liqing; Zhang, Jizong; Chen, Kaiyun E-mail: firstname.lastname@example.com; Hu, Liqun E-mail: email@example.com; Li, Erzhong; Lin, Shiyao; Shi, Tonghui; Duan, Yanmin; Zhu, Yubao
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.
Neutral redox-active hydrogen- and halogen-bonding [2]rotaxanes for the electrochemical sensing of chloride.
Lim, Jason Y C; Cunningham, Matthew J; Davis, Jason J; Beer, Paul D
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.
A Highly Active and Robust Copper-Based Electrocatalyst toward Hydrogen Evolution Reaction with Low Overpotential in Neutral Solution.
Du, Jialei; Wang, Jianying; Ji, Lvlv; Xu, Xiaoxiang; Chen, Zuofeng
Although significant progress has been made recently, copper-based materials have long been considered to be ineffective catalysts toward the hydrogen evolution reaction (HER), in most cases, requiring high overpotentials more than 300 mV. We report here that a Cu(0)-based nanoparticle film electrodeposited in situ from a Cu(II) oxime complex can act as a highly active and robust HER electrocatalyst in neutral phosphate buffer solution. The as-prepared nanoparticle film is of poor crystallization, which incorporates significant amounts of oxime ligand residues and buffer anions PO4(3-). The proposed mechanism suggests that the Cu(0)-based nanoparticle film is activated with incorporated or adsorbed PO4(3-) anions and the PO4(3-) anions-anchored sites might serve as the actual catalytic active sites with efficient proton transport mediators. Catalysis occurs with a low onset overpotential (η) of 65 mV, and a current density of 1 mA/cm(2) can be achieved with η = 120 mV. The nanoparticle film shows an excellent catalytic durability with slightly rising current density during electrolysis, presumably due to further incorporation or adsorption of PO4(3-) anions in the process. This electrocatalyst not only forms in situ from earth-abundant materials but also operates in neutral water with low overpotential and high stability.
Specifically modified Env immunogens activate B-cell precursors of broadly neutralizing HIV-1 antibodies in transgenic mice.
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
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.
Specifically modified Env immunogens activate B-cell precursors of broadly neutralizing HIV-1 antibodies in transgenic mice
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
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
Evaluation of Shiga toxin 2e-specific chicken egg yolk immunoglobulin: production and neutralization activity.
Arimitsu, Hideyuki; Sasaki, Keiko; Kohda, Tomoko; Shimizu, Toshiyasu; Tsuji, Takao
Chicken egg yolk immunoglobulin (IgY) against Shiga toxin 2e (Stx2e), a major cause of swine edema disease, was prepared to evaluate its possible clinical applications. The titer of Stx2e-specific IgY in egg yolk derived from three chickens that had been immunized with an Stx2e toxoid increased 2 weeks after primary immunization and remained high until 90 days after this immunization. Anti-Stx2e IgY was found to neutralize the toxicity of Stx2e by reacting with its A and B subunits, indicating that IgY is a cost-effective agent to develop for prophylactic foods or diagnosis kits for edema disease.
Reinstatement of pain-related brain activation during the recognition of neutral images previously paired with nociceptive stimuli.
Forkmann, Katarina; Wiech, Katja; Sommer, Tobias; Bingel, Ulrike
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.
Neutralizing activity of antibodies to the V3 loop region of HIV-1 gp120 relative to their epitope fine specificity
Pantophlet, Ralph; Wrin, Terri; CavaciniJ, Lisa A.; Robinson, James E.; Burton, Dennis R.
The V3 loop of HIV-1 gp120 is considered occluded on many primary viruses. However, virus sensitivity to neutralization by different V3 mAbs often varies, indicating that access to V3 is not restricted equally for all antibodies. Here, we have sought to gain a better understanding of these restrictions by determining the neutralizing activities of 7 V3 mAbs (19b, 39F, CO11, F2A3, F530, LA21, and LE311) against 15 subtype B primary isolates and relating these activities to the fine specificity of the mAbs. Not surprisingly, we found that most mAbs neutralized the same 2–3 viruses, with only mAb F530 able to neutralize 2 additional viruses not neutralized by the other mAbs. Epitope mapping revealed that positively-charged residues in or near the V3 stem are important for the binding of all the mAbs and that most mAbs seem to require the Pro residue that forms the GPGR β hairpin turn in the V3 tip for binding. Based on the mapping, we determined that V3 sequence variation accounted for neutralization resistance of approximately half the viruses tested. Comparison of these results to those of select V3 mAbs with overall better neutralizing activities in the light of structural information suggests that an antibody's mode of interaction with V3, driven by contact residue requirements, precludes the antibody from accessing its epitope on different viruses. Based on the data we propose an angle of interaction with V3 that is less stringent on access for antibodies with cross-neutralizing activity compared to antibodies that neutralize relatively fewer viruses. PMID:18822440
Purification and characterization of neutral alpha-mannosidase from hen oviduct: studies on the activation mechanism of Co2+.
Yamashiro, K; Itoh, H; Yamagishi, M; Natsuka, S; Mega, T; Hase, S
Neutral alpha-mannosidase was purified to homogeneity from hen oviduct. The molecular mass of the enzyme was 480 kDa on gel filtration, and the 110-kDa band on SDS-PAGE in the presence of 2-mercaptoethanol indicated that it is composed of four subunits. The activated enzyme hydrolyzed both p-nitrophenyl alpha-D-mannoside and high mannose-type sugar chains. This substrate specificity is almost the same as that reported for the neutral a-mannosidase from Japanese quail oviduct [Oku and Hase (1991) J. Biochem. 110, 982-989]. Manalpha1-6(Manalpha1-3)Manalpha1-6(Manalpha1-3) Manbeta1-4GlcNAc (Km =0.44 mM) was hydrolyzed four times faster than Manalpha1-6(Manalpha1-3)Manalpha1-6(Manalpha1-3) Manbeta1-4GIcNAcbeta1-4GlcNAc, and Manalpha1-6(Manalpha1-2Manalpha1-2Manalpha1-3)++ +Manbeta1-4GlcNAc was obtained as the end product from Man9GlcNAc on digestion with the activated alpha-mannosidase. The enzyme was activated 24-fold on preincubation with Co2+. The activation with other metal ions, like Mn2+, Ca2+, Fe2+, Fe3+, and Sr2+, was less than 5-fold, and Zn2+, Cu2+, and Hg2+ inhibited the enzyme activity. The optimum pHs for both the enzyme activity and activation with Co2+ were around 7. The cobalt ion contents of the purified, EDTA-treated, and Co2+-activated enzymes were 1.5, 0.0, and 3.9, respectively, per molecule. Since the Co2+-activated enzyme gradually lost its activity on incubation with EDTA and the activity was restored promptly on the addition of Co2+, the binding of Co2+ to the enzyme seems to be essential for its activation. The results obtained with protease inhibitors together with those of the SDS-PAGE before and after activation, showed that the proteolytic cleavage reported for the activation of monkey brain alpha-mannosidase seems not to be involved.
Divergent Trends of Anti-JCPyV Serum Reactivity and Neutralizing Activity in Multiple Sclerosis (MS) Patients during Treatment with Natalizumab
Diotti, Roberta Antonia; Capra, Ruggero; Moiola, Lucia; Caputo, Valeria; De Rossi, Nicola; Sangalli, Francesca; Martinelli, Vittorio; Burioni, Roberto; Clementi, Massimo; Mancini, Nicasio
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
Effect of delayed anthrax vaccine dose on Bacillus anthracis protective antigen IgG response and lethal toxin neutralization activity.
Pittman, Phillip R; Fisher, Diana; Quinn, Xiaofei; Schmader, Trevor; Barrera-Oro, Julio G
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.
Neutron activation analysis system
Taylor, M.C.; Rhodes, J.R.
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)
Impact of diagnostic neutral beam optimization on active spectroscopy in MST
NASA Astrophysics Data System (ADS)
Feng, Xiande; Nornberg, Mark. D.; den Hartog, Daniel. J.; Oliva, Steven. P.; Craig, Darren; Univ of Wisconsin, Madison Team; Wheaton College, IL Collaboration
The 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 has been optimized to operate at 50kV, 4A steady beam current with 20ms beam pulse and 75% primary energy ion fraction. It's achieved by tuning the beam voltage, arc current, fuel line pressure, arc and high voltage module timing, and the magnetic isolation field. Electron density measurements in the ion source revealed that ion extraction is maximized under low density conditions which are thought to affect the shape of the ion sheath at the extraction grid. The sheath may be transitioning from a planar or convex shape at high density to one which is concave which helps focus the ion trajectories and produce higher beam current. With the improvements in beam operation, the CHERS signal is expected to increase by 20%-30%, and the Stark broadening is expected to increase by 10%. These signal increases will help resolve convolved fine-structure components in both analyses. Beam voltage ripple is also measured to better quantify the accuracy of spectral MSE and CHERS measurement. This work is supported by the U.S. DOE.
Cross-Neutralization Activity of Single-Chain Variable Fragment (scFv) Derived from Anti-V3 Monoclonal Antibodies Mediated by Post-Attachment Binding.
Maruta, Yasuhiro; Kuwata, Takeo; Tanaka, Kazuki; Alam, Muntasir; Valdez, Kristel Paola Ramirez; Egami, Yoshika; Suwa, Yoshiaki; Morioka, Hiroshi; Matsushita, Shuzo
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.
Opposite effects of EGTA and neutral surfactants on the loss of chicken pineal serotonin N-acetyltransferase activity.
Rodriguez-Cabello, J C; Arpa, F; Agapito, M T; Recio, J M
The effects of some general purpose drugs on the deactivation and activity measurement of the chicken pineal gland enzyme serotonin N-acetyl transferase (EC 184.108.40.206.) were studied. The drugs used were EGTA and two neutral surfactants, Nonidet P40 and Triton X-100. Enzyme activity showed significant variations ranging from 2.8 +/- 1.3 nmol/gland/h when Nonidet P40 was added to the homogenate buffer, to 31.8 +/- 1.7 nmol/gland/h when EGTA was present. This striking variation seemed to be caused by the ability of these compounds to modify the rate of NAT deactivation acting either as accelerating agents, as in the case of the detergents or as braking agent, as in the case of EGTA.
Improved volatile fatty acids anaerobic production from waste activated sludge by pH regulation: Alkaline or neutral pH?
Ma, Huijun; Chen, Xingchun; Liu, He; Liu, Hongbo; Fu, Bo
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.
Cryo-EM structures elucidate neutralizing mechanisms of anti-chikungunya human monoclonal antibodies with therapeutic activity
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.
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. In this paper, 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. Finally, 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.
Cryo-EM structures elucidate neutralizing mechanisms of anti-chikungunya human monoclonal antibodies with therapeutic activity
Long, Feng; Fong, Rachel H.; Austin, Stephen K.; ...
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. In this paper, 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 ofmore » 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. Finally, 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.« less
Analysis of heavy oils: Method development and application to Cerro Negro heavy petroleum: Preliminary separation and analysis of acid, base, saturate, and neutral-aromatic fractions
Green, J.B.; Sturm, G.P. Jr.; Reynolds, J.W.; Thomson, J.S.; Yu, S.K-T.; Grigsby, R.D.; Tang, S.Y.; Shay, J.Y.; Hirsch, D.E.; Sanchez, V.
Cerro Negro 200-425/degree/, 425-550/degree/, and 550-700/degree/C distillates and >700/degree/C residue were separated into acid, base, and neutral concentrates using an improved nonaqueous ion exchange liquid chromatographic procedure. Neutral concentrates were further separated into neutral aromatic and saturated hydrocarbon fractions. A dual column, normal phase high performance liquid chromatographic method was developed for the saturate-aromatic separation. Mass and elemental balances are given for separations of all distillates and the residue. In addition, fractions from the 200-425/degree/C and 425-550/degree/C distillates were analyzed by high resolution mass spectrometry. The applicability of published separation approaches and methods to heavy oil analysis is critically reviewed; the bulk of the available methodology developed for conventional petroleum and synfuels was found to be unproven or unsuitable for heavy oil analysis. Cerro Negro was found to contain 18.2 weight percent acids, 17.6 weight percent bases, 46.9 weight percent neutral aromatics, and 14.7 weight percent saturated hydrocarbons. Saturate fractions contained predominantly cycloparaffins, neutral-aromatics were largely comprised of aromatic hydrocarbons and sulfur compounds, bases were largely nitrogen-containing compounds, and acids were mostly oxygen-containing compounds and nitrogen compounds of pyrrolic type. 145 refs., 24 figs., 21 tabs.
Heliopause Instabilities Driven by Plasma-Neutral Interaction: Linear Analysis from Fluid Model
Dasgupta, B.; Avinash, K.; Florinski, V.; Zank, G. P.; Heerikhuisen, J.; Bandyopadhyay, A.
The heliopause (HP) is the outer edge of the heliosphere that separates the tenuous and hot heliosheath plasma from the relatively dense and cool magnetized plasma of the local interstellar medium (LISM). As a surface of tangential discontinuity, the HP is subject to both Rayleigh-Taylor (RT) and Kelvin-Helmholtz (KH) instabilities. The coupling between plasma ions and neutral atoms through charge exchange provides an ''effective gravity'' for RT, while shear flows due to the velocity difference between the heliosheath and the interstellar flows can drive KH modes. We derive the linearized dispersion relations for waves propagating along the surface of this discontinuity using a fluid picture, to study the possibility for both RT and KH instabilities, including a second population of energetic neutrals produced by charge exchange interactions between the hot solar wind plasma in the heliosheath with interstellar hydrogen. We show that due to charge exchange interactions, the HP is unconditionally unstable to all perturbations.
Numerical Analysis of Neutral Entrainment Effect on Field-Reversed Configuration Thruster Efficiency
The flow conditions include neon gas and plasma with densities on the order of 1018 molecule⁄m3, plasma temperatures between 5 and 10 eV, and relative...configuration thrusters. The flow conditions include neon gas and plasma with densities on the order of 1018 molecule∕m3, plasma temperatures between 5...includes a planar flow between a field- reversed configuration plasmoid initially at a Schmid–Burgk equilibrium and neutral gas with varying relative
Neutralization of pharmacological and toxic activities of bothrops snake venoms by Schizolobium parahyba (Fabaceae) aqueous extract and its fractions.
Vale, Luis Henrique F; Mendes, Mirian M; Hamaguchi, Amélia; Soares, Andreimar M; Rodrigues, Veridiana M; Homsi-Brandeburgo, Maria Inês
The aqueous extract prepared from Schizolobium parahyba (Sp) leaves, a native plant from Atlantic Forest (Brazil), was tested to analyse its ability to inhibit some biological and enzymatic activities induced by Bothrops alternatus (BaltCV) and Bothrops moojeni (BmooCV) snake venoms. Sp inhibited 100% of lethality, blood incoagulability, haemorrhagic and indirect haemolytic activities at a 1:10 ratio (venom/extract, w/w), as well as coagulant activity at a 1:5 ratio (venom/extract, w/w) induced by both venoms. BaltCV fibrinogenolytic activity was also neutralized by Sp at a 1:10 ratio, resulting in total protection of fibrinogen Bbeta chain and partial protection of Aalpha chain. Interaction tests have demonstrated that, at certain extract/proteins ratios, Sp precipitates proteins non-specifically suggesting the presence of tannins, which are very likely responsible for the excellent inhibiting effects of the analysed ophidian activities. Sp aqueous extract chromatography on Sephadex LH-20 was carried out aiming at the separation of these compounds that mask the obtained results. Thus, the fractionation of Sp resulted in three fractions: F1 (methanolic fraction); F2 (methanol:water fraction, 1:1 v/v); and F3 (aqueous fraction). These fractions were analysed for their ability to inhibit the BaltCV fibrinogenolytic activity. F1 inhibited 100% the venom fibrinogenolytic activity without presenting protein precipitation effect; F2 showed only partial inhibition of this venom activity. Finally, F3 did not inhibit fibrinogen proteolysis, but presented strong protein precipitating action. We conclude that Sp aqueous extract, together with tannins, also contains other compounds that can display specific inhibitory activity against snake venom toxins.
The Anti-Tumor Activity of a Neutralizing Nanobody Targeting Leptin Receptor in a Mouse Model of Melanoma
McMurphy, Travis; Xiao, Run; Magee, Daniel; Slater, Andrew; Zabeau, Lennart; Tavernier, Jan; Cao, Lei
Environmental and genetic activation of a brain-adipocyte axis inhibits cancer progression. Leptin is the primary peripheral mediator of this anticancer effect in a mouse model of melanoma. In this study we assessed the effect of a leptin receptor antagonist on melanoma progression. Local administration of a neutralizing nanobody targeting the leptin receptor at low dose adjacent to tumor decreased tumor mass with no effects on body weight or food intake. In contrast, systemic administration of the nanobody failed to suppress tumor growth. Daily intraperitoneal injection of high-dose nanobody led to weight gain, hyperphagia, increased adiposity, hyperleptinemia, and hyperinsulinemia, and central effects mimicking leptin deficiency. The blockade of central actions of leptin by systemic delivery of nanobody may compromise its anticancer effect, underscoring the need to develop peripherally acting leptin antagonists coupled with efficient cancer-targeting delivery. PMID:24587106
Sorption of ionized and neutral emerging trace organic compounds onto activated sludge from different wastewater treatment configurations.
Hyland, Katherine C; Dickenson, Eric R V; Drewes, Jörg E; Higgins, Christopher P
The objective of this study was to examine sorption of a suite of 19 trace organic contaminants (TOrCs) to activated sludge. Compounds examined in this study included neutral, nonionized TOrCs as well as acidic TOrCs which may carry a negative charge and basic TOrCs which may carry a positive charge at the pH of wastewater. These TOrCs were evaluated to examine how sorptive behavior might differ for TOrCs in different states of charge. Additionally, multiple sludges from geographically and operationally different wastewater treatment plants were studied to elicit how solid-phase characteristics influence TOrC sorption. Characterization of sludge solids from 6 full scale treatment facilities and 3 bench-scale reactors showed no significant difference in fraction organic carbon (f(oc)) and cation exchange capacity (CEC). Sorption experiments demonstrated that sorption of TOrCs also exhibits little variation between these different sludges. Organic carbon normalized partition coefficients (logK(oc)) were determined as a measure of sorption, and were found to correlate well with octanol-water partition coefficients (logK(ow)) for nonionized TOrCs, and logD(ow) for anionic TOrCs where logD(ow) is greater than 2. These data were used to construct a linear free energy relationship (LFER), which was comparable to existing LFERs for sorption onto sludge. No trend in sorption was apparent for the remaining anionic TOrCs or for the cationic TOrCs. These data suggest that predicting sorption to activated sludge based on K(ow) values is a reasonable approach for neutral TOrCs using existing LFERs, but electrostatic (and likely other) interactions may govern the sorptive behavior of the charged organic chemicals to sludge.
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.
Activity of Ca2+-dependent neutral proteases in tissues of ground squirrel during hibernation and during self-warming after induced awakening.
Nurmagomedova, P M; Abasova, M M; Emirbekov, E Z
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.
Allicin from garlic neutralizes the hemolytic activity of intra- and extra-cellular pneumolysin O in vitro.
Arzanlou, M; Bohlooli, S; Jannati, E; Mirzanejad-Asl, H
Pneumolysin (PLY) is a key virulence factor contributes to the pathogenesis of Streptococcus pneumoniae. In this study we investigated the effect of allicin and aqueous garlic extracts on hemolytic activity of PLY both in prelysed and intact cells. Additionally the antimicrobial activity of allicin was tested against the bacteria. All tested materials potently inhibited the PLY hemolytic activity. Allicin neutralizes PLY in a concentration- and time-dependent manner. Twenty five minute incubation of PLY (2 HU/mL) with 0.61 μM/mL concentration of allicin, totally inhibited hemolytic activity of PLY (IC50 = 0.28 μM/mL). The inhibitory activity of old extract of garlic was similar to pure allicin (IC50 = 50.46 μL/mL; 0.31 μM/mL; P < 0.05). In contrast fresh extract of garlic inhibits the PLY hemolytic activity at lower concentrations (IC50 = 13.96 μL/mL; 0.08 μM/mL allicin). Exposure of intact cells to allicin (1.8 μM) completely inhibited hemolytic activity of PLY inside bacterial cells. The inhibitory effect of the allicin was restored by addition of reducing agent DTT at 5 mM, proposing that allicin likely inhibits the PLY by binding to cysteinyl residue in the binding site. The MIC value of allicin was determined to be 512 μg/mL (3.15 μM/mL). These results indicate that PLY is a novel target for allicin and may provide a new line of investigation on pneumococcal diseases in the future.
Neutral particle beam intensity controller
Dagenhart, William K.
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.
Enantioselective analysis of chiral anteiso fatty acids in the polar and neutral lipids of food.
Hauff, Simone; Hottinger, Georg; Vetter, Walter
Anteiso fatty acids (aFA) are substituted with a methyl group on the antepenultimate carbon of the straight acyl chain. This feature leads to a stereogenic center. The 12-methyltetradecanoic acid (a15:0) and the 14-methylhexadecanoic acid (a17:0) are the most common aFA found in food, although they occur only in very small quantities. In this study we used gas chromatography in combination with a chiral stationary phase to determine the enantiomeric distribution of both a15:0 and a17:0 in the neutral and polar lipids of aquatic food samples and cheese. The best suited column was selected out of four custom-made combinations of heptakis(6-O-tert-butyldimethylsilyl-2,3-di-O-methyl)-beta-cyclodextrin (beta-TBDM) with different amount and polarity of an achiral polysiloxane. After separation of polar and neutral lipids of the food samples by solid phase extraction, fatty acid methyl esters were prepared and the fatty acid methyl esters were fractionated by reversed phase high performance liquid chromatography. Measurements of fractions high in aFA by enantioselective GC/MS in the selected ion monitoring mode verified the dominance of the (S)-enantiomers of a15:0 and a17:0 in both lipid fractions. However (R)-enantiomers were detectable in all samples. The relative proportion of the (R)-enantiomers was up to fivefold higher in the polar lipids than in the neutral lipids. The higher proportions in the polar lipids indicate that microorganisms might be involved in the formation of (R)-aFA.
Design and evaluation of a hybrid passive and active gravity neutral orthosis (GNO).
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
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.
Neutral Lipids and Peroxisome Proliferator-Activated Receptor-γ Control Pulmonary Gene Expression and Inflammation-Triggered Pathogenesis in Lysosomal Acid Lipase Knockout Mice
Lian, Xuemei; Yan, Cong; Qin, Yulin; Knox, Lana; Li, Tingyu; Du, Hong
The functional roles of neutral lipids in the lung are poorly understood. However, blocking cholesteryl ester and triglyceride metabolism in lysosomal acid lipase gene knockout mice (lal−/−) results in severe pathogenic phenotypes in the lung, including massive neutrophil infiltration, foamy macrophage accumulation, unwanted cell growth, and emphysema. To elucidate the mechanism underlining these pathologies, we performed Affymetrix GeneChip microarray analysis of 1-, 3-, and 6-month-old mice and identified aberrant gene expression that progressed with age. Among changed genes, matrix metalloproteinase (MMP)-12, apoptosis inhibitor 6 (Api-6), erythroblast transformation-specific domain (Ets) transcription factor family member Spi-C, and oncogene MafB were increased 100-, 70-, 40-, and 10-fold, respectively, in lal−/− lungs versus the wild-type lungs. The pathogenic increases of these molecules occurred primarily in alveolar type II epithelial cells. Transcriptional activities of the MMP-12 and Api-6 promoters were stimulated by Spi-C or MafB in respiratory epithelial cells. Treatment with 9-hydroxyoctadecanoic acids and ciglitazone significantly rescued lal−/− pulmonary inflammation and aberrant gene expression. In addition, both compounds as well as peroxisome proliferator-activated receptor gamma inhibited MMP-12 and Api-6 promoter activities. These data suggest that inflammation-triggered cell growth and emphysema during lysosomal acid lipase deficiency are partially caused by peroxisome proliferator-activated receptor-γ inactivation. PMID:16127159
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
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.
Theoretical study of chlordecone and surface groups interaction in an activated carbon model under acidic and neutral conditions.
Gamboa-Carballo, Juan José; Melchor-Rodríguez, Kenia; Hernández-Valdés, Daniel; Enriquez-Victorero, Carlos; Montero-Alejo, Ana Lilian; Gaspard, Sarra; Jáuregui-Haza, Ulises Javier
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.
Mass analysis of neutral particles and ions released during electrical breakdowns on spacecraft surfaces
NASA Technical Reports Server (NTRS)
Kendall, B. R. F.
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.
Compact, accurate description of diagnostic neutral beam propagation and attenuation in a high temperature plasma for charge exchange recombination spectroscopy analysis.
Bespamyatnov, Igor O; Rowan, William L; Granetz, Robert S
Charge exchange recombination spectroscopy on Alcator C-Mod relies on the use of the diagnostic neutral beam injector as a source of neutral particles which penetrate deep into the plasma. It employs the emission resulting from the interaction of the beam atoms with fully ionized impurity ions. To interpret the emission from a given point in the plasma as the density of emitting impurity ions, the density of beam atoms must be known. Here, an analysis of beam propagation is described which yields the beam density profile throughout the beam trajectory from the neutral beam injector to the core of the plasma. The analysis includes the effects of beam formation, attenuation in the neutral gas surrounding the plasma, and attenuation in the plasma. In the course of this work, a numerical simulation and an analytical approximation for beam divergence are developed. The description is made sufficiently compact to yield accurate results in a time consistent with between-shot analysis.
Generation and characterization of tabalumab, a human monoclonal antibody that neutralizes both soluble and membrane-bound B-cell activating factor
Manetta, Joseph; Bina, Holly; Ryan, Paul; Fox, Niles; Witcher, Derrick R; Kikly, Kristine
B-cell activating factor (BAFF) is a B-cell survival factor with a key role in B-cell homeostasis and tolerance. Dysregulated BAFF expression may contribute to autoimmune diseases or B-cell malignancies via effects on abnormal B-lymphocyte activation, proliferation, survival, and immunoglobulin secretion. Monoclonal antibodies were generated against human BAFF, characterized for species specificity and affinity, and screened for the ability to neutralize both membrane-bound and soluble BAFF. In addition, studies were undertaken to determine the relative potency of membrane-bound and soluble BAFF. Tabalumab has a high affinity for human, cynomolgus monkey, and rabbit BAFF. No binding to mouse BAFF was detected. Tabalumab was able to neutralize soluble human, cynomolgus monkey, or rabbit BAFF with equal potency. Our data demonstrate that membrane-bound BAFF can be a more potent stimulus for B-cells than soluble BAFF, and tabalumab also neutralized membrane-bound BAFF. Tabalumab prevented BAFF from binding to BAFF receptors and demonstrated pharmacodynamic effects in human BAFF transgenic mice. Tabalumab is a high-affinity human antibody with neutralizing activity against membrane-bound and soluble BAFF. Given our findings that membrane-bound BAFF can have greater in vitro potency than soluble BAFF, neutralization of both forms of BAFF is likely to be important for optimal therapeutic effect. PMID:25258549
Complexity of Neutralizing Antibodies against Multiple Dengue Virus Serotypes after Heterotypic Immunization and Secondary Infection Revealed by In-Depth Analysis of Cross-Reactive Antibodies
Tsai, Wen-Yang; Durbin, Anna; Tsai, Jih-Jin; Hsieh, Szu-Chia; Whitehead, Stephen
ABSTRACT The four serotypes of dengue virus (DENV) cause the most important and rapidly emerging arboviral diseases in humans. The recent phase 2b and 3 studies of a tetravalent dengue vaccine reported a moderate efficacy despite the presence of neutralizing antibodies, highlighting the need for a better understanding of neutralizing antibodies in polyclonal human sera. Certain type-specific (TS) antibodies were recently discovered to account for the monotypic neutralizing activity and protection after primary DENV infection. The nature of neutralizing antibodies after secondary DENV infection remains largely unknown. In this study, we examined sera from 10 vaccinees with well-documented exposure to first and second DENV serotypes through heterotypic immunization with live-attenuated vaccines. Higher serum IgG avidities to both exposed and nonexposed serotypes were found after secondary immunization than after primary immunization. Using a two-step depletion protocol to remove different anti-envelope antibodies, including group-reactive (GR) and complex-reactive (CR) antibodies separately, we found GR and CR antibodies together contributed to more than 50% of neutralizing activities against multiple serotypes after secondary immunization. Similar findings were demonstrated in patients after secondary infection. Anti-envelope antibodies recognizing previously exposed serotypes consisted of a large proportion of GR antibodies, CR antibodies, and a small proportion of TS antibodies, whereas those recognizing nonexposed serotypes consisted of GR and CR antibodies. These findings have implications for sequential heterotypic immunization or primary immunization of DENV-primed individuals as alternative strategies for DENV vaccination. The complexity of neutralizing antibodies after secondary infection provides new insights into the difficulty of their application as surrogates of protection. IMPORTANCE The four serotypes of dengue virus (DENV) are the leading cause of
Exosomes neutralize synaptic-plasticity-disrupting activity of Aβ assemblies in vivo
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
Antioxidant activity of idebenone-loaded neutral and cationic solid-lipid nanoparticles.
Leonardi, Antonio; Crasci', Lucia; Panico, Annamaria; Pignatello, Rosario
Idebenone (IDE) is a lipophilic benzoquinone electron carrier synthetic analogue of coenzyme Q10, which behaves as an antioxidant and free radical scavenging molecule. Recently, the therapeutic application of IDE in Leber's hereditary optic neuropathy has been discussed. This work was aimed at evaluating the encapsulation of IDE in solid-lipid nanoparticles (SLN). In particular, we tested the possibility of adapting the quasi-emulsion solvent diffusion technique, already proposed to produce polymeric nanoparticles, to prepare positively charged SLN with different compositions. Such a charge, due to the addition of a cationic lipid, would facilitate the interaction with the negatively charged eye surface epithelium, with a consequent longer pre-corneal residence time of the colloidal systems. In a preliminary evaluation of the produced IDE-loaded SLN, the antioxidant activity of the drug was demonstrated using an oxygen radical absorbance capacity assay. Encapsulation of the drug in the nanocarrier systems seems able to protect IDE from degradation and prolong its antioxidant potential.
The neuromuscular activity of Micrurus pyrrhocryptus venom and its neutralization by commercial and specific coral snake antivenoms
Camargo, Thiago Magalhães; de Roodt, Adolfo Rafael; da Cruz-Höfling, Maria Alice; Rodrigues-Simioni, Léa
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
Approach on quantitative structure-activity relationship for design of a pH neutral carrier containing tertiary amino group.
Cao, Zhong; Gong, Fu-Chun; Li, He-Ping; Xiao, Zhong-Liang; Long, Shu; Zhang, Ling; Peng, San-Jun
The quantitative structure-activity relationship (QSAR) for neutral carriers used to prepare hydrogen ion sensors has been studied. A series of synthesized carrier compounds were taken as the training set. Five molecular structure parameters of the compounds were calculated by using CNDO/2 algorithm and used as feature variables in constructing QSAR model. The lower and upper limits of the linear pH response range were taken as the activity measure. The corresponding model equations were derived from the stepwise regression procedure. With the established QSAR model, a new pH carrier, (4-hydroxybenzyl) didodecylamine (XIII) was proposed and synthesized. The PVC membrane pH electrode based on carrier XIII with a wide pH linear response range of 2.0-12.5 was prepared. Having a theoretical Nernstian response slope of 57.2+/-0.3 mV/pH (n=5 at 25 degrees C) without a super-Nernstian phenomenon, the sensor had low resistance, short response time, high selectivity and good reproducibility. Moreover, the sensor was successfully applied to detecting the pH value of serum samples.
Bench-scale study of active mine water treatment using cement kiln dust (CKD) as a neutralization agent.
Mackie, Allison L; Walsh, Margaret E
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.
Simulation of Cl− Secretion in Epithelial Tissues: New Methodology Estimating Activity of Electro-Neutral Cl− Transporter
Sasamoto, Kouhei; Niisato, Naomi; Taruno, Akiyuki; Marunaka, Yoshinori
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
SPEAR 3 flight analysis: Grounding by neutral gas release, and magnetic field effects on current distribution
NASA Astrophysics Data System (ADS)
Mandell, M. J.; Jongeward, G. A.; Cooke, D. L.; Raitt, W. J.
The Space Power Experiment Aboard Rockets (SPEAR) 3 experiment was launched on March 15, 1993, to test grounding devices for negative payloads. In this paper we review two aspects of the high-altitude flight data and compare them with preflight predictions. The SPEAR 3 neutral gas release experiment studied a grounding mechanism observed on previous flights during attitude control system (ACS) firings. Preflight calculations using Paschen law physics generalized to three dimensions predicted that the high rate gas release (about one order of magnitude below normal ACS) would reduce the rocket potential to within 200-300 V of plasma ground. The flight data is well fit by a value of -225V. Orientation relative to Earth's magnetic field had no effect on the floating potential or grounding operations but had a large effect on the portion of the current collected by the boom. We compare these flight measurements with preflight calculations made with the DynaPAC computer code.
Analysis of secondary particle behavior in multiaperture, multigrid accelerator for the ITER neutral beam injector.
Mizuno, T; Taniguchi, M; Kashiwagi, M; Umeda, N; Tobari, H; Watanabe, K; Dairaku, M; Sakamoto, K; Inoue, T
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.
Ebola virus-like particles produced in insect cells exhibit dendritic cell stimulating activity and induce neutralizing antibodies
Ye Ling; Lin Jianguo; Sun Yuliang; Bennouna, Soumaya; Lo, Michael; Wu Qingyang; Bu Zhigao; Pulendran, Bali; Compans, Richard W. . E-mail: email@example.com; Yang Chinglai . E-mail: email@example.com
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.
Expression patterns, activities and carbohydrate-metabolizing regulation of sucrose phosphate synthase, sucrose synthase and neutral invertase in pineapple fruit during development and ripening.
Zhang, Xiu-Mei; Wang, Wei; Du, Li-Qing; Xie, Jiang-Hui; Yao, Yan-Li; Sun, Guang-Ming
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.
Expression Patterns, Activities and Carbohydrate-Metabolizing Regulation of Sucrose Phosphate Synthase, Sucrose Synthase and Neutral Invertase in Pineapple Fruit during Development and Ripening
Zhang, Xiu-Mei; Wang, Wei; Du, Li-Qing; Xie, Jiang-Hui; Yao, Yan-Li; Sun, Guang-Ming
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. PMID:22949808
Evolutionary neutrality of mtDNA introgression: evidence from complete mitogenome analysis in roe deer.
Matosiuk, M; Sheremetyeva, I N; Sheremetyev, I S; Saveljev, A P; Borkowska, A
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.
Decanting of Neutralized H-Canyon Unirradiated Nuclear Material High Activity Waste Streams
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
Direct analysis of sialylated or sulfated glycosphingolipids and other polar and neutral lipids using TLC-MS interfaces.
Park, Hyejung; Zhou, Ying; Costello, Catherine E
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.
Changes in EEG activity before and after exhaustive exercise in sedentary women in neutral and hot environments.
Ftaiti, Foued; Kacem, Asma; Jaidane, Nadia; Tabka, Zouhair; Dogui, Mohamed
This study examined the effect of hyperthermia on brain electrical activity measured with encephalography during prolonged exhaustive exercise in a group of sedentary women (VO(2)max = 35 +/- 4 mL kg min(-1)). Two strenuous cycling exercises were performed either in neutral (N-Ex) or in heat (H-Ex) conditions. Tympanic temperature (Tty), heart rate (HR), body mass loss (BML), plasma volume decrease, and brain electrical activity [EEG: alpha (8-13 Hz) and beta(13-30 Hz)-band and alpha/beta index of fatigue: the ratio between EEG activity in the alpha band and beta-band] were recorded throughout the cycling sessions. The Tty increase 1.0 degrees C in the N-Ex and 1.8 degrees C in H-Ex. HR increased in both sessions but with significantly higher values during the H-Ex session when compared with the N-Ex session (p < 0.001) (from 85 +/- 4 beats min(-1) to 164 +/- 6 beats min(-1) and from 83 +/- 6 beats min(-1) to 181 +/- 8 beats min(-1), respectively in N-Ex and in H-Ex). This was associated with a significantly higher BML (p < 0.05) and a higher plasma volume decrease in the H-Ex session (p < 0.01). The alpha/beta index increased significantly during both trials particularly during the H-Ex session (p < 0.05). This was associated with a significant decrease of time to exhaustion (-34%). We suggest that exhausting work in the heat induced a change in gross brain activity (alpha/beta ratio) compared to a longer, less thermally demanding exposure. Fatigue in the heat could be attributed to central factors as well as thermal, cardiac and hydro-electrolytic impairment.
Cell selectivity, mechanism of action and LPS-neutralizing activity of bovine myeloid antimicrobial peptide-18 (BMAP-18) and its analogs.
Lee, Eun Kyu; Kim, Yoon-Chang; Nan, Yong Hai; Shin, Song Yub
To develop novel antimicrobial peptides (AMPs) with improved cell selectivity and potent LPS-neutralizing activity, we synthesized an 18 N-terminal residues peptide (BAMP-18) of bovine myeloid antimicrobial peptide-27 (BMAP-27) and its analogs (BMAP-18-W, BMAP-18-L, BMAP-18-I and BMAP-18-f). BMAP-18 and its analogs displayed much higher cell selectivity (about 4-97-fold increased) as compared to parental BMAP-27 because of their decreased hemolytic activity and retained antimicrobial activity. BMAP-27 caused near-complete dye leakage from bacterial-membrane-mimicking vesicles even at very low concentration of 0.5μM, whereas BMAP-18 and its analogs induced very little dye leakage (less than 40%) even at 16μM. These peptides induced near-complete membrane depolarization of Staphylococcus aureus cells under their MIC (4μM). These results suggests that BMAP-18 and its analogs exhibit lethality toward microbes due to their ability to form small channels that permit the transit of ions or protons, but not molecules as large as calcein, and not by the membrane-disruption/perturbation mode. BMAP-18 and its analogs significantly inhibited nitric oxide (NO) production or tumor necrosis factor-α (TNF-α) release in LPS-stimulated mouse macrophage RAW264.7 cells at 10μM. In particular, BMAP-18-W showed LPS-neutralizing activity comparable to that of BMAP-27. There was a significant linear correlation between the increase in the hydrophobicity of peptides and LPS-neutralizing activity. Although BMAP-18-W has lower hydrophobicity than BMAP-18-L, it showed higher LPS-neutralizing activity as compared to BMAP-18-L. This result suggests other important parameters of AMPs may be involved in their LPS-neutralizing activity, as well as positive charge and hydrophobicity.
A highly active and stable palladium catalyst on a g-C3N4 support for direct formic acid synthesis under neutral conditions.
Park, Hunmin; Lee, Ju Hyung; Kim, Eun Hyup; Kim, Kwang Young; Choi, Yo Han; Youn, Duck Hyun; Lee, Jae Sung
Graphitic carbon nitride (g-C3N4) is applied as a support of the Pd catalyst for direct HCOOH synthesis by CO2 hydrogenation under neutral conditions. The high CO2 affinity of g-C3N4 is responsible for the enhanced catalytic activity and stability relative to the inert support such as a carbon nanotube.
Oxalates as Activating Groups for Alcohols in Visible Light Photoredox Catalysis: Formation of Quaternary Centers by Redox-Neutral Fragment Coupling
MacMillan, David W. C.; Overman, Larry E.
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
Structure, Aggregation, and Activity of a Covalent Insulin Dimer Formed During Storage of Neutral Formulation of Human Insulin.
Hjorth, Christian Fogt; Norrman, Mathias; Wahlund, Per-Olof; Benie, Andrew J; Petersen, Bent O; Jessen, Christian M; Pedersen, Thomas Å; Vestergaard, Kirsten; Steensgaard, Dorte B; Pedersen, Jan Skov; Naver, Helle; Hubálek, František; Poulsen, Christian; Otzen, Daniel
A specific covalently linked dimeric species of insulin high molecular weight products (HMWPs), formed during prolonged incubation of a neutral pharmaceutical formulation of human insulin, were characterized in terms of tertiary structure, self-association, biological activity, and fibrillation properties. The dimer was formed by a covalent link between A21Asn and B29Lys. It was analyzed using static and dynamic light scattering and small-angle X-ray scattering to evaluate its self-association behavior. The tertiary structure was obtained using nuclear magnetic resonance and X-ray crystallography. The biological activity of HMWP was determined using 2 in vitro assays, and its influence on fibrillation was investigated using Thioflavin T assays. The dimer's tertiary structure was nearly identical to that of the noncovalent insulin dimer, and it was able to form hexamers in the presence of zinc. The dimer exhibited reduced propensity for self-association in the absence of zinc but significantly postponed the onset of fibrillation in insulin formulations. Consistent with its dimeric state, the tested species of HMWP showed little to no biological activity in the used assays. This study is the first detailed characterization of a specific type of human insulin HMWP formed during storage of a marketed pharmaceutical formulation. These results indicate that this specific type of HMWP is unlikely to antagonize the physical stability of the formulation, as HMWP retained a tertiary structure similar to the noncovalent dimer and participated in hexamer assembly in the presence of zinc. In addition, increasing amounts of HMWP reduce the rate of insulin fibrillation.
Learning the Relationship between the Primary Structure of HIV Envelope Glycoproteins and Neutralization Activity of Particular Antibodies by Using Artificial Neural Networks
Buiu, Cătălin; Putz, Mihai V.; Avram, Speranta
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
Nonneutralizing Antibodies Induced by the HIV-1 gp41 NHR Domain Gain Neutralizing Activity in the Presence of the HIV Fusion Inhibitor Enfuvirtide: a Potential Therapeutic Vaccine Strategy.
Wang, Qian; Bi, Wenwen; Zhu, Xiaojie; Li, Haoyang; Qi, Qianqian; Yu, Fei; Lu, Lu; Jiang, Shibo
A key barrier against developing preventive and therapeutic human immunodeficiency virus (HIV) vaccines is the inability of viral envelope glycoproteins to elicit broad and potent neutralizing antibodies. However, in the presence of fusion inhibitor enfuvirtide, we show that the nonneutralizing antibodies induced by the HIV type 1 (HIV-1) gp41 N-terminal heptad repeat (NHR) domain (N63) exhibit potent and broad neutralizing activity against laboratory-adapted HIV-1 strains, including the drug-resistant variants, and primary HIV-1 isolates with different subtypes, suggesting the potential of developing gp41-targeted HIV therapeutic vaccines.
Structural characterization of neutral oligosaccharides with blood-group A and H activity isolated from bovine submaxillary mucin.
Savage, A V; D'Arcy, S M; Donoghue, C M
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
Neutral "Cp-Free" Silyl-Lanthanide(II) Complexes: Synthesis, Structure, and Bonding Analysis
Complexes featuring lanthanide silicon bonds represent a research area still in its infancy. Herein, we report a series of Cp-free lanthanide (+II) complexes bearing σ-bonded silyl ligands. By reactions of LnI2 (Ln = Yb, Eu, Sm) either with a 1,4-oligosilanyl dianion [K-Si(SiMe3)2SiMe2SiMe2Si(SiMe3)2-K)] (1) or with 2 (Me3Si)3SiK (3) the corresponding neutral metallacyclopentasilanes ({Me2Si(Me3Si)2Si}2)Ln·(THF)4 (Ln = Yb (2a), Eu (2b), Sm (2c)), or the disilylated complexes ({Me3Si}3Si)2Ln·(THF)3 (Ln = Yb (4a), Eu (4b), Sm (4c)), were selectively obtained. Complexes 2b, 2c, 4b, and 4c represent the first examples of structurally characterized Cp-free Eu and Sm complexes with silyl ligands. In both series, a linear correlation was observed between the Ln–Si bond lengths and the covalent radii of the corresponding lanthanide metals. Density functional theory calculations were also carried out for complexes 2a–c and 4a–c to elucidate the bonding situation between the Ln(+II) centers and Si. PMID:26132550
Neutralization Analysis of a Chicken Single-Chain Variable Fragment Derived from an Immune Antibody Library Against Infectious Bronchitis Virus.
Lin, Yuan; Li, Benqiang; Ye, Jiaxin; Wang, Man; Wang, Jianhua; Zhang, Ying; Zhu, Jianguo
Avian infectious bronchitis virus (IBV), which is prevalent in many countries causing severe economic loss to the poultry industry, causes infectious bronchitis (IB) in birds. Recombinant single-chain variable fragments (scFvs) have been proven to effectively inhibit many viruses, both in vitro and in vivo, and they could be a potential diagnostic and therapeutic reagent to control IB. In this study, six anti-IBV chicken scFvs, ZL.10, ZL.64, ZL.78, ZL.80, ZL.138, and ZL.256, were obtained by screening random clones from an immune antibody library. An analysis of nucleotide sequences revealed that they represented distinctive genetic sequences and greatly varied in complementarity-determining region three of the heavy chain. Neutralization tests showed that ZL.10, which bound the S1 protein in western blots, inhibited the formation of syncytia in Vero cells 48 h post IBV infection and decreased the transcriptional level of nucleoprotein mRNA to 17.2%, while the other five scFvs, including ZL.78 and ZL.256, that bound the N protein did not. In conclusion, the results suggested that specific and neutralizing chicken scFvs against IBV, which can be safe and economical antibody reagents, can be produced in vitro through prokaryotic expression.
Neutralizing Antibodies Protect against Lethal Flavivirus Challenge but Allow for the Development of Active Humoral Immunity to a Nonstructural Virus Protein
Kreil, Thomas R.; Maier, Elisabeth; Fraiss, Sabine; Eibl, Martha M.
Antibody-mediated neutralization of viruses has been extensively studied in vitro, but the precise mechanisms that account for antibody-mediated protection against viral infection in vivo still remain largely uncharacterized. The two points under discussion are antibodies conferring sterilizing immunity by neutralizing the virus inoculum or protection against the development of disease without complete inhibition of virus replication. For tick-borne encephalitis virus (TBEV), a flavivirus, transfer of neutralizing antibodies specific for envelope glycoprotein E protected mice from subsequent TBEV challenge. Nevertheless, short-term, low-level virus replication was detected in these mice. Furthermore, mice that were exposed to replicating but not to inactivated virus while passively protected developed active immunity to TBEV rechallenge. Despite the priming of TBEV-specific cytotoxic T cells, adoptive transfer of serum but not of T cells conferred immunity upon naive recipient mice. These transferred sera were not neutralizing and were predominantly specific for NS1, a nonstructural TBEV protein which is expressed in and on infected cells and which is also secreted from these cells. Results of these experiments showed that despite passive protection by neutralizing antibodies, limited virus replication occurs, indicating protection from disease rather than sterilizing immunity. The protective immunity induced by replicating virus is surprisingly not T-cell mediated but is due to antibodies against a nonstructural virus protein absent from the virion. PMID:9525632
Monoclonal Antibody Combinations that Present Synergistic Neutralizing Activity: A Platform for Next-Generation Anti-Toxin Drugs
Diamant, Eran; Torgeman, Amram; Ozeri, Eyal; Zichel, Ran
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
The early melting of closed duplex DNA: analysis by banding in buoyant neutral rubidium trichloroacetate.
Burke, R L; Bauer, W R
Aqueous RbTCA permits the buoyant banding of both native and denatured DNA at room temperature and neutral pH. A unique property of this solvent is the bouyant resolution of closed circular, underwound DNA (I) from the corresponding nicked (II) species. Conditions are reported here in which PM-2 DNA I is physically resolved from native PM-2 DNA II, the buoyant separation being 1.27 mq/ml in 3.3 M RbTCA at 25 degrees C. The separation between nicked and closed DNAs increases with temperature up to 35.5 degrees C, at which PM-2 DNA II cooperatively melts and subsequently pellets. The isothermal buoyant density of a cloed DNA increases linearly as the linking number (Lk) of the closed DNA decreases. The early melting of closed DNA may be monitored with high precision by buoyant banding in RbTCA, it being possible to detect the disruption of as few as 40 base pairs in PM-2 DNA (10,000 base pairs). The constraint that the linking number be conserved in closed DNA requires that a change in duplex winding be accompanied by a compensating change in supercoiling. We estimate the linking number deficiency of PM-2 DNA I to be 0.094 turns per decibase pair. This result permits the estimation of the EtdBr unwinding angle, phi, by comparison with alternative determinations of the linking number deficiency which depend upom the value of phi. The result obtained here is that phi = 27.7 degrees +/- 0.5 degrees and is approximately independent of temperature over the range 15 degrees-35 degrees. PMID:7443544
NLTE Analysis of High-Resolution H-band Spectra. I. Neutral Silicon
NASA Astrophysics Data System (ADS)
Zhang, Junbo; Shi, Jianrong; Pan, Kaike; Allende Prieto, Carlos; Liu, Chao
We investigated the reliability of our silicon atomic model and the influence of non-local thermodynamical equilibrium (NLTE) on the formation of neutral silicon (Si i) lines in the near-infrared (near-IR) H-band. We derived the differential Si abundances for 13 sample stars with high-resolution H-band spectra from the Apache Point Observatory Galactic Evolution Experiment (APOGEE), as well as from optical spectra, both under local thermodynamical equilibrium (LTE) and NLTE conditions. We found that the differences between the Si abundances derived from the H-band and from optical lines for the same stars are less than 0.1 dex when the NLTE effects are included, and that NLTE reduces the line-to-line scatter in the H-band spectra for most sample stars. These results suggest that our Si atomic model is appropriate for studying the formation of H-band Si lines. Our calculations show that the NLTE corrections of the Si i H-band lines are negative, i.e., the final Si abundances will be overestimated in LTE. The corrections for strong lines depend on surface gravity, and tend to be larger for giants, reaching ˜-0.2 dex in our sample, and up to ˜-0.4 dex in extreme cases of APOGEE targets. Thus, the NLTE effects should be included in deriving silicon abundances from H-band Si i lines, especially for the cases where only strong lines are available. Based on observations collected with the 2.16 m telescope at Xinglong station, National Astronomical Observatories, Chinese Academy of Sciences, the 2.2 m telescope at the Calar Alto Observatory, the 1.88 m reflector at the Okayama Astrophysical Observatory, the Kitt Peak coudé feed telescope, and the McMath-Pierce solar telescope and the coudé focus of the Mayall 4 m reflector at Kitt Peak.
Analysis and application of a neutralizing linear epitope on liable toxin B of enterotoxin Escherichia coli.
Guan, Weikun; Liu, Wenxin; Bao, Jun; Li, Jinping; Yuan, Chaowen; Tang, Jie; Shi, Dongfang
Heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli (ETEC) is one of the major virulence factors for causing diarrhea in piglets, and LT is a strong immunogen. Thus, LT represents an important target for development of vaccines and diagnostic tests. In this study, bioinformatic tools were used to predict six antigenic B cell epitopes in the B subunit of LT protein (LTB) of ETEC strains. Then, seven antigenic B cell epitopes of LTB were identified by polyclonal antisera (polyclonal antibody (PAb)) using a set of LTB-derived peptides expressed as maltose-binding protein (MBP) fusion protein. In addition, one LTB-specific monoclonal antibody (MAb) was generated and defined its corresponding epitope as mentioned above. This MAb was able to specifically bind with native LT toxin and has no cross-reaction with LT-II (type II heat-labile enterotoxin), Stx1 (Shiga toxin I), Stx2 (Shiga toxin II), STa (heat-stable enterotoxin I), and STb (heat-stable enterotoxin II) toxins. Further, this MAb was able to interrupt LT toxin specific binding to GM1 receptor, indicating that the corresponding epitope is the specific binding region to GM1 receptor. Moreover, in vitro and in vivo assay showed that the MAb was able to neutralize the native LT toxin. Diarrheal suckling pigs challenged with LT-positive ETEC strain recovered when an enema with this purified MAb was administered. This study will provide the foundation for further studies about the interaction between LT toxin and GM1 receptor and about the developing of epitope-based vaccines and specific therapeutic agent.
A pulmonary influenza virus infection in SCID mice can be cured by treatment with hemagglutinin-specific antibodies that display very low virus-neutralizing activity in vitro.
Mozdzanowska, K; Furchner, M; Washko, G; Mozdzanowski, J; Gerhard, W
We have previously shown that a pulmonary influenza virus infection in SCID mice can be cured by treatment with monoclonal antibodies (MAbs) specific for the viral transmembrane protein hemagglutinin (HA) but not for matrix 2. Since both types of MAbs react with infected cells but only the former neutralizes the virus, it appeared that passive MAbs cured by neutralization of progeny virus rather than reaction with infected host cells. To prove this, we selected a set of four HA-specific MAbs, all of the immunoglobulin G2a isotype, which reacted well with native HA expressed on infected cells yet differed greatly (>10,000-fold) in virus neutralization (VN) activity in vitro, apparently because of differences in antibody avidity and accessibility of the respective determinants on the HA of mature virions. Since the VN activities of these MAbs in vitro were differentially enhanced by serum components, we determined their prophylactic activities in vivo and used them as measures of their actual VN activities in vivo. The comparison of therapeutic and prophylactic activities indicated that these MAbs cured the infection to a greater extent by VN activity (which was greatly enhanced in vivo) and to a lesser extent by reaction with infected host cells. Neither complement- nor NK cell-dependent mechanisms were involved in the MAb-mediated virus clearance. PMID:9151823
Hyperimmune antisera against synthetic peptides representing the glycoprotein of human immunodeficiency virus type 2 can mediate neutralization and antibody-dependent cytotoxic activity.
Björling, E; Broliden, K; Bernardi, D; Utter, G; Thorstensson, R; Chiodi, F; Norrby, E
Twenty-five 13- to 35-amino-acid-long peptides representing regions of human immunodeficiency virus type 2 (HIV-2), strain SBL6669, envelope proteins were evaluated for their immunogenic activity in guinea pigs. The peptides were selected to provide homologous representation of sites in the HIV-1 envelope proteins that were previously documented to have a particular immunogenic importance. A number of the HIV-2 peptides were found to be capable of inducing strain SBL6669 neutralizing and antibody-dependent cellular cytotoxicity (ADCC) antibodies. Two overlapping peptides covering amino acids 311-337 representing the central and C-terminal part of the variable third (V3) region, terminology according to Modrow et al. [Modrow, S., Hahn, B., Shaw, G. M., Gallo, R. C., Wong-Staal, F. & Wolf, H. (1987) J. Virol. 61, 570-578], showed the most pronounced capacity to induce neutralizing antibodies. One of the peptides (amino acids 318-337) also induced antibodies mediating ADCC. Two additional regions in the large glycoprotein, gp125, containing linear sites reacting with neutralizing antibodies were identified (amino acids, 119-137 and 472-509). The transmembrane protein, gp36, of HIV-2 harbored two regions of importance for induction of neutralizing antibodies (amino acids 595-614 and 714-729). ADCC activity was induced by two additional gp125-specific peptides (amino acids 291-311 and 446-461). Thus, except for the single V3-specific site there was no correlation between linear immunogenic sites stimulating neutralizing antibody and ADCC activity. These findings pave the way for development of synthetic vaccines against HIV-2 and possibly also simian immunodeficiency virus infections. The capacity of such a product to induce protective immunity can be evaluated in macaque monkeys. Images PMID:2068087
Are there physical links between Saturn's magnetospheric planetary period oscillations, neutral atmosphere circulation, and thunderstorm activity? (Invited)
NASA Astrophysics Data System (ADS)
Provan, G.; Cowley, S. W.
Suggestions that the planetary period oscillations (PPOs) observed in Saturn's magnetosphere may be driven or influenced by neutral atmospheric perturbations, motivates an exploratory comparison of PPO rotation periods with available tropospheric and stratospheric determinations. Non-polar atmospheric rotation periods occupy the range ~10.2-10.7 h associated with the latitudinal jet structure, are similar north and south, and independent of season, while PPO periods lie in a narrower partly overlapping range ~10.6-10.8 h, are persistently shorter north than south, and undergo a seasonal cycle. In this cycle, widely-separated north-south PPO periods during southern summer converge across equinox to values lying within the atmospheric west jet band, remaining well-separated from east jet periods. Closest convergence occurred one year post-equinox, contemporaneously with the switch in seasonal thunderstorm activity from southern to northern hemispheres. Since most large-scale atmospheric phenomena are related to the west jets, rotating with closely similar periods, they also rotate with periods close to the PPOs under post-equinoctial conditions, but not otherwise. Specifically, post-equinox northern PPOs rotate with a period close to the southern thunderstorms, as well as the north polar spot and hexagon features, while the post equinox southern PPOs rotate with a period close to the pre-equinox northern ';string of pearls' and the first co-located post-equinox northern thunderstorm, the Great White Spot event. However, even under these conditions no consistent correspondences in period are found at a detailed level, which taken together with the lack of correspondence at other times, does not suggest a direct physical link exists between these phenomena.
Saturn's magnetospheric planetary period oscillations, neutral atmosphere circulation, and thunderstorm activity: Implications, or otherwise, for physical links
NASA Astrophysics Data System (ADS)
Cowley, S. W. H.; Provan, G.
that the planetary period oscillations (PPOs) observed in Saturn's magnetosphere may be driven or influenced by neutral atmospheric perturbations motivate an exploratory comparison of PPO rotation periods with available tropospheric and stratospheric determinations. Nonpolar atmospheric rotation periods occupy the range ~10.2-10.7 h associated with the latitudinal jet structure, are similar north and south, and are independent of season, while PPO periods lie in a narrower partly overlapping range ~10.6-10.8 h, are persistently shorter north than south, and undergo a seasonal cycle. In this cycle, widely separated north-south PPO periods during southern summer converge across equinox to values lying within the atmospheric west jet band, remaining well-separated from east jet periods. Closest convergence occurred 1 year post equinox, contemporaneously with the switch in seasonal thunderstorm activity from Southern to Northern Hemispheres. Since most large-scale atmospheric phenomena are related to the west jets, rotating with closely similar periods, they also rotate with periods close to the PPOs under post equinoctial conditions but not otherwise. Specifically, post equinox northern PPOs rotate with a period close to the southern thunderstorms, as well as the north polar spot and hexagon features, while the post equinox southern PPOs rotate with a period close to the pre-equinox northern "string of pearls" and the first colocated post equinox northern thunderstorm, the Great White Spot event. However, even under these conditions, no consistent correspondences in period are found at a detailed level, which taken together with the lack of correspondence at other times does not suggest a direct physical link exists between these phenomena.
Crystal Structure of PG16 and Chimeric Dissection with Somatically Related PG9: Structure-Function Analysis of Two Quaternary-Specific Antibodies That Effectively Neutralize HIV-1
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.
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.
Toward Understanding the Cold, Hot, and Neutral Nature of Chinese Medicines Using in Silico Mode-of-Action Analysis.
Fu, Xianjun; Mervin, Lewis H; Li, Xuebo; Yu, Huayun; Li, Jiaoyang; Mohamad Zobir, Siti Zuraidah; Zoufir, Azedine; Zhou, Yang; Song, Yongmei; Wang, Zhenguo; Bender, Andreas
One important, however, poorly understood, concept of Traditional Chinese Medicine (TCM) is that of hot, cold, and neutral nature of its bioactive principles. To advance the field, in this study, we analyzed compound-nature pairs from TCM on a large scale (>23 000 structures) via chemical space visualizations to understand its physicochemical domain and in silico target prediction to understand differences related to their modes-of-action (MoA) against proteins. We found that overall TCM natures spread into different subclusters with specific molecular patterns, as opposed to forming coherent global groups. Compounds associated with cold nature had a lower clogP and contain more aliphatic rings than the other groups and were found to control detoxification, heat-clearing, heart development processes, and have sedative function, associated with "Mental and behavioural disorders" diseases. While compounds associated with hot nature were on average of lower molecular weight, have more aromatic ring systems than other groups, frequently seemed to control body temperature, have cardio-protection function, improve fertility and sexual function, and represent excitatory or activating effects, associated with "endocrine, nutritional and metabolic diseases" and "diseases of the circulatory system". Compounds associated with neutral nature had a higher polar surface area and contain more cyclohexene moieties than other groups and seem to be related to memory function, suggesting that their nature may be a useful guide for their utility in neural degenerative diseases. We were hence able to elucidate the difference between different nature classes in TCM on the molecular level, and on a large data set, for the first time, thereby helping a better understanding of TCM nature theory and bridging the gap between traditional medicine and our current understanding of the human body.
Appreciation of Hardy's Poem "Neutral Tones" from the Point of View of Functional Analysis
ERIC Educational Resources Information Center
Chen, Hong-qin
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…
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
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…
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.
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
The neutral lipid composition present in the digestive vacuole of Plasmodium falciparum concentrates heme and mediates β-hematin formation with an unusually low activation energy.
Hoang, Anh N; Sandlin, Rebecca D; Omar, Aneesa; Egan, Timothy J; Wright, David W
In eukaryotic cells, neutral lipids serve as major energy storage molecules; however, in Plasmodium falciparum, a parasite responsible for causing malaria in humans, neutral lipids may have other functions during the intraerythrocytic stage of the parasite life cycle. Specifically, experimental data suggest that neutral lipid structures behave as a catalyst for the crystallization of hemozoin, a detoxification byproduct of several blood-feeding organisms, including malaria parasites. Synthetic neutral lipid droplets (SNLDs) were produced by depositing a lipid blend solution comprised of mono- and diglycerides onto an aqueous surface. These lipid droplets are able to mediate the production of brown pigments that are morphologically and chemically identical to hemozoin. The partitioning of heme into these SNLDs was examined by employing Nile Red, a lipid specific dye. Soluble ferriprotoporphyrin IX was observed to spontaneously localize to the lipid droplets, partitioning in a pH-dependent manner with an estimated log P of 2.6. Interestingly, the pH profile of heme partitioning closely resembles that of β-hematin formation. Differential scanning calorimetry and kinetic studies demonstrated that the SNLDs provide a unique environment that promotes hemozoin formation. SNLD-mediated formation of the malaria pigment displayed an activation energy barrier lower than those of individual lipid components. In particular, lipid droplets composed of diglycerides displayed activation barriers lower than those composed of monoglycerides. This difference was attributed to the greater fluidity of these lipids. In conjunction with the known pattern of lipid body proliferation, it is suggested that neutral lipid structures within the digestive vacuole not only are the location of in vivo hemozoin formation but are also essential for the survival of the parasite by functioning as a kinetically competent and site specific mediator for heme detoxification.
Juliano, Steven A
Competition experiments estimating the relative effects of inter- and intraspecific competition can help to resolve whether interspecific competition results in coexistence or exclusion. For mosquitoes, most such experiments have focused on invasive Aedes albopictus and its interactions with resident Aedes. A meta-analysis of such experiments tested whether the effect of interspecific competition is greater than, less than, or equal to that of intraspecific competition, and whether competitive outcomes are dependent on food quality. For A. albopictus and A. aegypti, there was significant context dependence, with interspecific competitive advantage for A. albopictus with low food quality, and competitive equivalence with high food quality. Meta-analysis of survivorship yielded more significant effects than did estimated rate of increase. Competitive effects and competitive responses of each species yielded similar results. This meta-analysis suggests competitive exclusion of A. aegypti by A. albopictus, and is thus consistent with field sampling, qualitative reviews, and interpretations from individual publications. For A. albopictus and A. triseriatus, most results indicated competitive equivalence and no context dependence, and are thus contrary to previous qualitative reviews and to interpretations from individual publications. For both pairs of species, published results suitable for meta-analysis remain scarce, and better experimental designs and improved analysis and reporting of statistical results are needed. Greater emphasis needs to be placed on estimating species' inter- and intra-specific competitive effects, rather than the more common, but theoretically less interesting, competitive responses. Experiments without low-density controls (i.e., replacement series) are inadequate for comparing competitive effects and responses.
Juliano, Steven A.
Competition experiments estimating the relative effects of inter- and intraspecific competition can help to resolve whether interspecific competition results in coexistence or exclusion. For mosquitoes, most such experiments have focused on invasive Aedes albopictus and its interactions with resident Aedes. A meta-analysis of such experiments tested whether the effect of interspecific competition is greater than, less than, or equal to that of intraspecific competition, and whether competitive outcomes are dependent on food quality. For A. albopictus and A. aegypti, there was significant context dependence, with interspecific competitive advantage for A. albopictus with low food quality, and competitive equivalence with high food quality. Meta-analysis of survivorship yielded more significant effects than did estimated rate of increase. Competitive effects and competitive responses of each species yielded similar results. This meta-analysis suggests competitive exclusion of A. aegypti by A. albopictus, and is thus consistent with field sampling, qualitative reviews, and interpretations from individual publications. For A. albopictus and A. triseriatus, most results indicated competitive equivalence and no context dependence, and are thus contrary to previous qualitative reviews and to interpretations from individual publications. For both pairs of species, published results suitable for meta-analysis remain scarce, and better experimental designs and improved analysis and reporting of statistical results are needed. Greater emphasis needs to be placed on estimating species' inter- and intra-specific competitive effects, rather than the more common, but theoretically less interesting, competitive responses. Experiments without low-density controls (i.e., replacement series) are inadequate for comparing competitive effects and responses. PMID:23482823
The Activity of Neutral α-Glucosidase and Selected Biochemical Parameters in the Annual Cycle of Breeding Carp (Cyprinus carpio L.).
Kotońska-Feiga, Julia; Dobicki, Wojciech; Pokorny, Przemysław; Nowacki, Wojciech
The aim of the study was to demonstrate seasonal changes in the hydrolytic and transferase activity of neutral α-glucosidase, the level of glucose, cholesterol, triglycerides and total protein in the annual breeding cycle of the carp. The study was conducted on fish from a fish farm in Lower Silesia (Poland). Blood serum was collected from the heart in: June, September and December of two consecutive years. The results of the study show that the hydrolytic and transferase activity of neutral α-glucosidase, as well as the results of basic biochemical parameters are highest in summer, when the fish seek and intake food intensively. The lowest values were observed in spring, when carp have the lowest metabolism after the wintering period.
Neutralized nanoparticle composed of SS-cleavable and pH-activated lipid-like material as a long-lasting and liver-specific gene delivery system.
Ukawa, Masami; Akita, Hidetaka; Hayashi, Yasuhiro; Ishiba, Ryohei; Tange, Kota; Arai, Masaya; Kubo, Kazuhiro; Higuchi, Yuriko; Shimizu, Kazunori; Konishi, Satoshi; Hashida, Mitsuru; Harashima, Hideyoshi
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.
Plasma pharmacokinetics and biological activity of a human immunodeficiency virus type 1 neutralizing human monoclonal antibody, F105, in cynomolgus monkeys.
Cavacini, L A; Power, J; Emes, C L; Mace, K; Treacy, G; Posner, M R
The IgG1 kappa human monoclonal antibody (HMab), F105 reacts with a discontinuous epitope on the CD4 binding site (CD4BS) of human immunodeficiency virus type 1 (HIV-1)/gp120 and has broad neutralizing activity. F105 HMab (60 mg/kg bolus) was administered intravenously to four monkeys and serum was collected at intervals to determine pharmacokinetics in a primate model. Average serum F105 concentrations, as determined by enzyme-linked immunosorbent assay, were analyzed with MINSQ software using a two-compartment, first-order model. The half-life for the alpha phase of the distribution curve is 6.7 h and for the beta elimination phase, 9.6 days. The volume of distribution is 0.65 L/kg and the rate of clearance 2 ml/kg/h. Serum levels of 1.3-1.6 mg/ml of F105 were maintained for 24 h. When monkey serum from day 15 postdose was tested, total serum F105 was 230 +/- 79 micrograms/ml and was immunoreactive with cells infected with the MN and IIIB strains of HIV-1 as determined by flow cytometry. Binding activity was identical to that obtained with stock F105 HMab. Identical neutralizing activity between the injected and uninjected antibody was also observed. Thus, serum neutralizing titers (90%) of 1:2000 at peak and 1:30 at day 15 postdose for MN virus were observed. These data indicate that high in vivo levels of HMab F105 can be attained by single bolus administration with full retention of biological activity. Of importance, levels of antibody necessary for effective neutralization can be achieved and maintained.
Kinetic analysis about the effects of neutral salts on the thermal stability of yeast alcohol dehydrogenase.
Ikegaya, Kazuo
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
Numerical analysis of the angular motion of a neutrally buoyant spheroid in shear flow at small Reynolds numbers.
Rosén, T; Einarsson, J; Nordmark, A; Aidun, C K; Lundell, F; Mehlig, B
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.
Quetiapine Inhibits Microglial Activation by Neutralizing Abnormal STIM1-Mediated Intercellular Calcium Homeostasis and Promotes Myelin Repair in a Cuprizone-Induced Mouse Model of Demyelination
Wang, Hanzhi; Liu, Shubao; Tian, Yanping; Wu, Xiyan; He, Yangtao; Li, Chengren; Namaka, Michael; Kong, Jiming; Li, Hongli; Xiao, Lan
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
Surface analysis of rubbers and plastics using secondary neutral mass spectrometry.
Pellin, M. J.; Savina, M. R.
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.
Constraining the Europa Neutral Torus
NASA Astrophysics Data System (ADS)
Smith, Howard T.; Mitchell, Donald; mauk, Barry; Johnson, Robert E.; clark, george
"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.
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.
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...
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...
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
B Spurrier; J Sampson; M Totrov; H Li; T ONeal; C Williams; J Robinson; M Gorny; S Zolla-Pazner; X Kong
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.
In vitro genotoxicity of neutral red after photo-activation and metabolic activation in the Ames test, the micronucleus test and the comet assay.
Guérard, Melanie; Zeller, Andreas; Singer, Thomas; Gocke, Elmar
Neutral red (Nr) is relatively non-toxic and is widely used as indicator dye in many biological test systems. It absorbs visible light and is known to act as a photosensitizer, involving the generation of reactive oxygen species (type-I reaction) and singlet oxygen (type-II reaction). The mutagenicity of Nr was determined in the Ames test (with Salmonella typhimurium strains TA1535, TA97, TA98, TA98NR, TA100, and TA102) with and without metabolic activation, and with and without photo-activation on agar plates. Similarly to the situation following metabolic activation, photo-mutagenicity of Nr was seen with all Salmonella strains tested, albeit with different effects between these strains. To our knowledge, Nr is the only photo-mutagen showing such a broad action. Since the effects are also observed in strains not known to be responsive to ROS, this indicates that ROS production is not the sole mode of action that leads to photo-genotoxicity. The reactive species produced by irradiation are short-lived as pre-irradiation of an Nr solution did not produce mutagenic effects when added to the bacteria. In addition, mutagenicity in TA98 following irradiation was stronger than in the nitroreductase-deficient strain TA98NR, indicating that nitro derivatives that are transformed by bacterial nitroreductase to hydroxylamines appear to play a role in the photo-mutagenicity of Nr. Photo-genotoxicity of Nr was further investigated in the comet assay and micronucleus test in L5178Y cells. Concentration-dependent increases in primary DNA damage and in the frequency of micronuclei were observed after irradiation.
Search for active neutrino disappearance using neutral-current interactions in the MINOS long-baseline experiment
Adamson, P.; Andreopoulos, C.; Arms, Kregg E.; Armstrong, R.; Auty, D.J.; Ayres, D.S.; Backhouse, C. Baller, B.; Barr, G.; Barrett, W.L.; Becker, B.R.; /Minnesota U. /Rutherford
We have measured the rates and spectra of neutral-current neutrino interactions in the MINOS detectors, which are separated by 734 km. A depletion in the rate at the far site would indicate mixing between {nu}{sub {mu}} and a sterile particle. The depletion of the total neutral-current event rate at the far site is limited to be below 17% at 90% confidence level without {nu}{sub e} appearance. Assuming oscillations occur at a single mass-squared splitting, a fit to the neutral- and charged-current energy spectra shows the fraction of {nu}{sub {mu}} oscillating to a sterile neutrino is 0.28{sub -0.28}{sup +0.25}(stat.+syst.). Including {nu}{sub e} appearance at the current experimental upper bound limits the depletion to be below 21% at 90% confidence level and the fit fraction of {nu}{sub {mu}} oscillating to a sterile neutrino is 0.43{sub -0.27}{sup +0.23}(stat.+syst.).
Analysis of neutral nitromusks in incenses by capillary electrophoresis in organic solvents and gas chromatography-mass spectrometry.
Gotti, Roberto; Fiori, Jessica; Mancini, Francesca; Cavrini, Vanni
Nitromusks used as fragrances in a variety of personal-care products, cleansers, and domestic deodorants, including incense sticks, are neutral nitro aromatic compounds; some of these have been reported as photosensitizers. In this work, their analysis was performed by capillary electrophoresis (CE) in a methanol-based background electrolyte (BGE). In particular, a 10 mM solution of citric acid in methanol was used; under these conditions the strong suppression of the electroosmotic flow favored the use of negatively charged surfactants as additives for the anodic migration of the studied analytes. To this end, sodium taurodeoxycholate (TDC) was supplemented at high concentration (190 mM) to the organic background electrolyte (BGE), showing strong indication of the ability to give micelle-like aggregates. Since nitromusks are characterized by the presence of a nitroaromatic ring with low charge density, their association with TDC aggregates can be ascribed to donor-acceptor interactions. Separation of musk xylene, musk ketone, and the banned musk moskene and musk ambrette was obtained under full nonaqueous BGE; the addition of relatively small water percentages (15% v/v) was found to be useful in improving the separation of pairs of adjacent peaks. Under optimized conditions (190 mM sodium TDC in methanol-water, 85-15 v/v containing citric acid 10 mM) the system was applied to the analysis of nitromusks in incense sticks extracted with methanol. The results were compared with those obtained by the analysis of the same samples using gas chromatography with mass detector. The expected different selectivity of separation obtained using the two techniques can be useful in the unambiguous determination of the analytes; furthermore, a substantial accord of the preliminary quantitative results achieved with the two methods was assumed as the confirmation of the potential reliability of CE performed with high percentage of organic solvent.
Lewis, George K; Pazgier, Marzena; Evans, David; Ferrari, Guido; Bournazos, Stylianos; Parsons, Matthew S; Bernard, Nicole F; Finzi, Andrés
It has been known for more than 30 years that Human Immunodeficiency Virus 1 (HIV-1) infection drives a very potent B cell response resulting in the production of anti-HIV-1 antibodies targeting several viral proteins, particularly its envelope glycoproteins (Env). Env epitopes are exposed on the surfaces of viral particles and infected cells where they are targets of potentially protective antibodies. These antibodies can interdict infection by neutralization and there is strong evidence suggesting that Fc-mediated effector function can also contribute to protection. Current evidence suggests that Fc-mediated effector function plays a role in protection against infection by broadly neutralizing antibodies (bnAbs) and it might be important for protection by non-neutralizing antibodies. Fc-mediated effector function includes diverse mechanisms that include antibody-dependent cellular cytotoxicity (ADCC), antibody-mediated complement activation (ADC), antibody-dependent cellular phagocytosis (ADCP), antibody-dependent cell-mediated virus inhibition (ADCVI), antibody-mediated trancytosis inhibition, and antibody-mediated virus opsonization. All these functions could be beneficial in fighting viral infections including HIV-1. In this perspective, we discuss the latest developments for ADCC responses discussed at the HIVR4P satellite session on non-neutralizing antibodies, with emphasis on the mechanisms of ADCC resistance employed by HIV-1, the structural basis of epitopes recognized by antibodies that mediate ADCC, NK-cell education and ADCC, and murine models to study ADCC against HIV-1.
The Double Helicity Asymmetry in Neutral Pion Production at PHENIX and Its Constraint on δG Through Global Analysis
NASA Astrophysics Data System (ADS)
Boyle, Kieran
Understanding the gluon spin contribution to the proton spin, δG, is a primary goal of the RHIC spin program. The abundant production of pions in p+p collisions, coupled with the PHENIX detector's fine resolution Electromagnetic Calorimeter and high pT triggering capabilities, make neutral pions a prime candidate to study the proton spin structure in polarized p+p collisions at RHIC. Measurements of the double helicity asymmetry, ALL, in ^0 production from 2005 and 2006 were shown to significantly constrain the gluon helicity distribution, δG, in the proton [1]. With improved luminosity and polarization, the figure of merit for the 2009 data set was a factor of 1.5 better that the previous runs combined. We present the 2009 results for ^0 ALL, along with a discussion of the results from inclusion of these data in a recent update of the DSSV global analysis.[4pt] [1] D. de Florian, R. Sassot, M. Stratmann, W. Vogelsang (DSSV) Phys.Rev.Lett. 101 (2008) 072001
Spatial Variations of Turbulent Properties in Neutral Hydrogen Observations of the Small Magellanic Cloud Using Structure Function Analysis
NASA Astrophysics Data System (ADS)
Nestingen-Palm, David; Stanimirovic, Snezana; Babler, Brian L.; Gonzalez Casanova, Diego; Jameson, Katherine; Bolatto, Alberto D.
Turbulence is known to play a key role in shaping the many structures and features which make the Interstellar Medium (ISM) so interesting to study. It is still not understood, however, which processes are responsible for the turbulence in the ISM, and the scales at which these processes dominate. In our study, we use the structure function to analyze neutral hydrogen (HI) observations of the Small Magellanic Cloud (SMC) to search for spacial variations in turbulent properties. Using an estimate of the star formation surface density as a guide, we separate the HI SMC observations into a central region which contains star formation rate (SFR) regions above a certain contour level, and an outer region which has SFR values below the chosen contour level. Our contour level begins at 1e-3 M⊙yr-1kpc-2 and is raised to 5e-3 M⊙yr-1kpc-2 by increments of 1e-4 M⊙yr-1kpc-2. At each contour level, we calculate the HI structure function slope for both the central and outer region by applying the Velocity Channel Analysis (VCA) and progressively averaging HI velocity channels. Our preliminary results suggest a difference in the slopes of both velocity and density fields between the inner and outer SMC.
On the biochemical classification of yeast trehalases: Candida albicans contains two enzymes with mixed features of neutral and acid trehalase activities.
Sánchez-Fresneda, Ruth; González-Párraga, Pilar; Esteban, Oscar; Laforet, Leslie; Valentín, Eulogio; Argüelles, Juan-Carlos
Two enzymes endowed with trehalase activity are present in Candida albicans. The cytosolic trehalase (Ntc1p), displayed high activity in exponential phase regardless of the carbon source (glucose, trehalose or glycerol). Ntc1p activity was similar in neutral (pH 7.1) or acid (pH 4.5) conditions, strongly inhibited by ATP, weakly stimulated by divalent cations (Ca(2+)or Mn(2+)) and unaffected in the presence of cyclic AMP. The Ntc1p activity decreased in stationary phase, except in glycerol-grown cultures, but the catalytic properties did not change. In turn, the cell wall-linked trehalase (Atc1p) showed elevated activity in resting cells or in cultures growing on trehalose or glycerol. Although Atc1p is subjected to glucose repression, exhaustion of glucose in itself did not increased the activity. Significant Atc1p values could also be measured at neutral or acid pH, but Atc1p was insensitive to ATP, cyclic AMP and divalent cations. These results are in direct contrast with the current classification of yeast trehalases based on their optimum pH. They are also relevant in the light of the proposed use of trehalase inhibitors for the treatment of candidiasis.
FeP nanoparticles film grown on carbon cloth: an ultrahighly active 3D hydrogen evolution cathode in both acidic and neutral solutions.
Tian, Jingqi; Liu, Qian; Liang, Yanhui; Xing, Zhicai; Asiri, Abdullah M; Sun, Xuping
In this Letter, we demonstrate the direct growth of FeP nanoparticles film on carbon cloth (FeP/CC) through low-temperature phosphidation of its Fe3O4/CC precursor. Remarkably, when used as an integrated 3D hydrogen evolution cathode, this FeP/CC electrode exhibits ultrahigh catalytic activity comparable to commercial Pt/C and good stability in acidic media. This electrode also performs well in neutral solutions. This work offers us the most cost-effective and active 3D cathode toward electrochemical water splitting for large-scale hydrogen fuel production.
Analysis of cross-reactive neutralizing antibodies in human HFMD serum with an EV71 pseudovirus-based assay.
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
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.
Mucosal Targeting of a BoNT/A Subunit Vaccine Adjuvanted with a Mast Cell Activator Enhances Induction of BoNT/A Neutralizing Antibodies in Rabbits
Staats, Herman F.; Fielhauer, Jeffrey R.; Thompson, Afton L.; Tripp, Alice A.; Sobel, Ashley E.; Maddaloni, Massimo; Abraham, Soman N.; Pascual, David W.
Background We previously reported that the immunogenicity of Hcβtre, a botulinum neurotoxin A (BoNT/A) immunogen, was enhanced by fusion to an epithelial cell binding domain, Ad2F, when nasally delivered to mice with cholera toxin (CT). This study was performed to determine if Ad2F would enhance the nasal immunogenicity of Hcβtre in rabbits, an animal model with a nasal cavity anatomy similar to humans. Since CT is not safe for human use, we also tested the adjuvant activity of compound 48/80 (C48/80), a mast cell activating compound previously determined to safely exhibit nasal adjuvant activity in mice. Methods New Zealand White or Dutch Belted rabbits were nasally immunized with Hcβtre or Hcβtre-Ad2F alone or combined with CT or C48/80, and serum samples were tested for the presence of Hcβtre-specific binding (ELISA) or BoNT/A neutralizing antibodies. Results Hcβtre-Ad2F nasally administered with CT induced serum anti-Hcβtre IgG ELISA and BoNT/A neutralizing antibody titers greater than those induced by Hcβtre + CT. C48/80 provided significant nasal adjuvant activity and induced BoNT/A-neutralizing antibodies similar to those induced by CT. Conclusions Ad2F enhanced the nasal immunogenicity of Hcβtre, and the mast cell activator C48/80 was an effective adjuvant for nasal immunization in rabbits, an animal model with a nasal cavity anatomy similar to that in humans. PMID:21304600
Structural Characterization of Humanized Nanobodies with Neutralizing Activity against the Bordetella pertussis CyaA-Hemolysin: Implications for a Potential Epitope of Toxin-Protective Antigen
Malik, Aijaz Ahmad; Imtong, Chompounoot; Sookrung, Nitat; Katzenmeier, Gerd; Chaicumpa, Wanpen; Angsuthanasombat, Chanan
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
Cytoskeletal protein Flightless (Flii) is elevated in chronic and acute human wounds and wound fluid: neutralizing its activity in chronic but not acute wound fluid improves cellular proliferation.
Ruzehaji, Nadira; Grose, Randall; Krumbiegel, Doreen; Zola, Heddy; Dasari, Pallave; Wallace, Hilary; Stacey, Michael; Fitridge, Robert; Cowin, Allison J
Chronic non-healing wounds form a medical need which will expand as the population ages and the obesity epidemic grows. Whilst the complex mechanisms underlying wound repair are not fully understood, remodelling of the actin cytoskeleton plays a critical role. Elevated expression of the actin cytoskeletal protein Flightless I (Flii) is known to impair wound outcomes. To determine if Flii is involved in the impaired healing observed in chronic wounds, its expression in non-healing human wounds from patients with venous leg ulcers was determined and compared to its expression in acute wounds and unwounded skin. Increased expression of Flii was observed in both chronic and acute wounds with wound fluid and plasma also containing secreted Flii protein. Inflammation is a key aspect of wound repair and fluorescence-activated cell sorting (FACS) analysis revealed Flii was located in neutrophils within the blood and that it co-localised with CD16+ neutrophils in chronic wounds. The function of secreted Flii was investigated as both chronic wound fluid and Flii have previously been shown to inhibit fibroblast proliferation. To determine if the inhibitory effect of wound fluid was due in part to the presence of Flii, wound fluids were depleted of Flii using Flii-specific neutralizing antibodies (FnAb). Flii depleted chronic wound fluid no longer inhibited fibroblast proliferation, suggesting that Flii may contribute to the inhibitory effect of chronic wound fluid on fibroblast function. Application of FnAbs to chronic wounds may therefore be a novel approach used to improve the local environment of non-healing wounds and potentially improve healing outcomes.
The cellular resistance against oxidative stress (H2O2) is independent of neutral trehalase (Ntc1p) activity in Candida albicans.
Pendreño, Yolanda; Pedreño, Y; González-Párraga, Pilar; González-Párraga, P; Conesa, Sergio; Conesa, S; Martínez-Esparza, María; Martínez-Esparza, M; Aguinaga, Ana; Aguinaga, A; Hernández, José A; Hernández, J A; Argüelles, Juan Carlos
The protective role of trehalose against oxidative stress caused by hydrogen peroxide in Candida albicans has been investigated in the homozygous mutant ntc1Delta/ntc1Delta, disrupted in the NTC1 gene, which encodes the neutral (cytosolic) trehalase (Ntc1p). After a severe oxidative exposure (50 mM H(2)O(2)), both parental (CAI-4) and ntc1Delta/ntc1Delta exponential-phase cells stored large amounts of intracellular trehalose. In turn, the degree of cell survival was roughly equivalent in both strains, although slightly higher in ntc1Delta/ntc1Delta cultures. The mechanism of 'adaptive tolerance' was functional in the two strains. Thus, a gently oxidative pretreatment (5 mM H(2)O(2)) increased the recovery of cellular viability when it was followed by a severe challenge (50 mM H(2)O(2)); this phenomenon was accompanied by a significant elevation of the endogenous trehalose content. Oxidative stress also induced specific activation of the antioxidant enzymes catalase and glutathione reductase upon gentle oxidative treatment (5 mM H(2)O(2)), whereas superoxide dismutase activity was only activated upon prolonged exposure. Taken together, these results strongly suggest that in C. albicans neutral trehalase activity does not play an essential role in the protective response against oxidative stress. They also suggest that a diminished Ntc1p activity might favour the growth of C. albicans cells subjected to a strong oxidative exposure.
Biological activity of neutral and cationic iridium(III) complexes with κP and κP,κS coordinated Ph₂PCH₂S(O)xPh (x = 0-2) ligands.
Ludwig, Gerd; Mijatović, Sanja; Ranđelović, Ivan; Bulatović, Mirna; Miljković, Djordje; Maksimović-Ivanić, Danijela; Korb, Marcus; Lang, Heinrich; Steinborn, Dirk; Kaluđerović, Goran N
Neutral iridium(III) complexes of the type [Ir(η(5)-C₅Me₅)Cl₂{Ph₂PCH₂S(O)xPh-κP}] (1-3) with diphenylphosphino-functionalized methyl phenyl sulfides, sulfoxides, and sulfones Ph₂PCH₂S(O)xPh (x = 0, L1; 1, L2; 2, L3) and the cationic complex [Ir(η(5)-C₅Me₅)Cl{Ph₂PCH₂SPh-κP,κS}][PF6] (4) were synthesized and fully characterized analytically and spectroscopically. Furthermore, the structure of 2 was determined by X-ray diffraction analysis. The biological potential of the neutral and cationic iridium(III) complexes was tested in vitro against the cell lines 8505C, A253, MCF-7, SW480 and 518A2. Complex [Ir(η(5)-C₅Me₅)Cl₂{Ph₂PCH₂S(O)Ph-κP}] (2), with ligand L2 κP coordinated containing a pendent sulfinyl group, is the most active one (IC₅₀ values of about 3 μM), thus, with activities comparable to cisplatin. Complex 2 proved to have an even a higher antiproliferative activity than cisplatin against 8505C and SW480 cell lines, used as a model system of highly anaplastic cancers with low sensitivity to conventional chemotherapeutics such as cisplatin. Additional experiments demonstrated that apoptosis and autophagic cell death contribute to the drug's tumoricidal action.
Neutralizing activity and protective immunity to ricin toxin conferred by B subunit (RTB)-specific Fab fragments.
Yermakova, Anastasiya; Mantis, Nicholas J
SylH3 and 24B11 are murine monoclonal antibodies directed against different epitopes on ricin toxin's binding (RTB) subunit that have been shown to passively protect mice against ricin challenge. Here we report that Fab fragments of SylH3 and 24B11 neutralize ricin in a cell based assay, and in a mouse challenge model as effectively as their respective full length parental IgGs. These data demonstrate that immunity to ricin can occur independent of Fc-mediated clearance.
Human Immunodeficiency Virus Type 1 gp120 Induces Apoptosis in Human Primary Neurons through Redox-Regulated Activation of Neutral Sphingomyelinase
Jana, Arundhati; Pahan, Kalipada
Human immunodeficiency virus type 1 (HIV-1) infection is known to cause disorders of the CNS, including HIV-associated dementia (HAD). HIV-1 coat protein gp120 (glycoprotein 120) induces neuronal apoptosis and has been implicated in the pathogenesis of HAD. However, the mechanism by which gp120 causes neuronal apoptosis is poorly understood. The present study underlines the importance of gp120 in inducing the production of ceramide, an important inducer of apoptosis, in human primary neurons. gp120 induced the activation of sphingomyelinases (primarily the neutral one) and the production of ceramide in primary neurons. Antisense knockdown of neutral (NSMase) but not acidic (ASMase) sphingomyelinase markedly inhibited gp120-mediated apoptosis and cell death of primary neurons, suggesting that the activation of NSMase but not ASMase plays an important role in gp120-mediated neuronal apoptosis. Similarly, the HIV-1 regulatory protein Tat also induced neuronal cell death via NSMase. Furthermore, gp120-induced production of ceramide was redox sensitive, because reactive oxygen species were involved in the activation of NSMase but not ASMase. gp120 coupled CXCR4 (CXC chemokine receptor 4) to induce NADPH oxidase-mediated production of superoxide radicals in neurons, which was involved in the activation of NSMase but not ASMase. These studies suggest that gp120 may induce neuronal apoptosis in the CNS of HAD patients through the CXCR4–NADPH oxidase–superoxide–NSMase–ceramide pathway. PMID:15509740
Effects of ethylene glycol tetraacetic acid, A23187 and calmodulin, calcium activated neutral proteinase antagonists on melatonin secretion in perifused chick pineal gland.
Agapito, M T; Pablos, M; Reiter, R J; Recio, J M; Gutierrez-Baraja, R
We have recently described, using perifused pineal glands, that calcium influx participates in the activation of chick pineal gland. This study shows that the loss of perifused chick pineal gland activity is a complex process which seems to involve the release of calcium from intracellular stores, calmodulin and calcium-activated neutral protease (CANP). Pineal glands were perifused with Krebs medium (controls) or with Krebs medium plus the drugs ethylene glycol tetraacetic acid (EGTA; calcium chelator), A23187 (calcium ionophore), EGTA plus A23187 (extra-intra cellular calcium chelation), trifluoperazine and CGS9343B (calmodulin inhibitors), and E-64 (CANP inhibitor) at the time of the natural peak of melatonin release. When EGTA or A23187 were added to the perifusion medium, no effects were observed. On the other hand, when the calcium chelator EGTA plus A23187 (free extra and intracellular calcium levels were dramatically decreased), trifluoperazine, CGS 9343B or E-64 were added to the perifusion medium melatonin synthesis increased significantly and was sustained for 8 h. We propose a prominent role for calcium output from intracellular stores in regulating melatonin production primarily by acting on Ca-calmodulin and calcium-activated neutral protease.
Characterization of the neutralizing activity of three anti-human TNF monoclonal antibodies and prediction of their TNF epitopes by molecular modeling and mutant protein approach.
Zhu, Cansheng; Liu, Xuesong; Feng, Jiannan; Zhang, Wei; Shen, Beifen; Ou'yang, Weiming; Cao, Yunxin; Jin, Boquan
The neutralizing activity of three anti-human TNF monoclonal antibodies, designated D2, E6, and F6 were investigated by three experimental systems. The results from the systems showed that all the three mAbs could neutralize TNF-mediated cytotoxicity in L929 cells, TNF-induced NF-kappaB activation in ECV304 cells, and TNF-upregulated ICAM-1 surface expression on ECV304 cells in dose-dependent manners. D2 had the highest neutralizing activity of the three mAbs, and F6 had higher level of neutralizing activity than E6. We also cloned the VH and VL cDNAs and obtained their cDNA sequences. The sequences were used in molecular modeling to establish the complex structures of TNF with variable regions of the three mAbs, respectively. In the structures, the TNF epitopes of D2, E6, and F6 were predicted at amino acids of (A109, A111-A112, C19, C21-C29, C44-C46, C66-C75, C77, C79, C90, C101, C103, C105, C114, C134-C148), (C18-C19, C21-C30, C32, C37, C43-C47, C67-C75, C83, C105-C106, C131, C135-C141), and (C21-C32, C45-C47, C65, C67-C72, C74, C81, C83, C90-C95, C105-C113, C133-C147), respectively, and the affinities of D2, E6, and F6 to TNF were predicted as -252.69, -232.83, and -299.92 kcal, respectively. Moreover, we proved the binding ability of F6 to the epitopes of amino acids of 141-146 in TNF molecule was better than that of E6, and that of D2 was the best of the three mAbs by Western blot and ELISA, in which the mutant TNF deleted the amino acids of 141-146 in TNF molecule was employed. These results make a basic foundation for selecting candidate mAbs for various purposes, such as construction of chimeric or humanized mAbs for therapeutic purpose, establishment of ELISA kits for determination of TNF, and production of affinity columns to purify TNF.
Second generation of pseudotype-based serum neutralization assay for Nipah virus antibodies: sensitive and high-throughput analysis utilizing secreted alkaline phosphatase.
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
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.
Nondestructive neutron activation analysis of volcanic samples: Hawaii
Zoller, W.H.; Finnegan, D.L.; Crowe, B.
Samples of volcanic emissions have been collected between and during eruptions of both Kilauea and Mauna Loa volcanoes during the last three years. Airborne particles have been collected on Teflon filters and acidic gases on base-impregnated cellulose filters. Chemically neutral gas-phase species are collected on charcoal-coated cellulose filters. The primary analytical technique used is nondestructive neutron activation analysis, which has been used to determine the quantities of up to 35 elements on the different filters. The use of neutron activation analysis makes it possible to analyze for a wide range of elements in the different matrices used for the collection and to learn about the distribution between particles and gas phases for each of the elements.
Analysis of the Pipe Heat Loss of the Water Flow Calorimetry System in EAST Neutral Beam Injector
NASA Astrophysics Data System (ADS)
Hu, Chundong; Chen, Yu; Xu, Yongjian; Yu, Ling; Li, Xiang; Zhang, Weitang
Neutral beam injection heating is one of the main auxiliary heating methods in controllable nuclear fusion research. In the EAST neutral beam injector, a water flow calorimetry (WFC) system is applied to measure the heat load on the electrode system of the ion source and the heat loading components of the beamline. Due to the heat loss in the return water pipe, there are some measuring errors for the current WFC system. In this paper, the errors were measured experimentally and analyzed theoretically, which lay a basis for the exact calculation of beam power deposition distribution and neutralization efficiency. supported by the National Magnetic Confinement Fusion Science Program of China (No. 2013GB101001) and the International Science & Technology Cooperation Program of China (No. 2014DFG61950)
9G4 autoreactivity is increased in HIV-infected patients and correlates with HIV broadly neutralizing serum activity.
Kobie, James J; Alcena, Danielle C; Zheng, Bo; Bryk, Peter; Mattiacio, Jonelle L; Brewer, Matthew; Labranche, Celia; Young, Faith M; Dewhurst, Stephen; Montefiori, David C; Rosenberg, Alexander F; Feng, Changyong; Jin, Xia; Keefer, Michael C; Sanz, Ignacio
The induction of a broadly neutralizing antibody (BNAb) response against HIV-1 would be a desirable feature of a protective vaccine. Vaccine strategies thus far have failed to elicit broadly neutralizing antibody responses; however a minority of HIV-infected patients do develop circulating BNAbs, from which several potent broadly neutralizing monoclonal antibodies (mAbs) have been isolated. The findings that several BNmAbs exhibit autoreactivity and that autoreactive serum antibodies are observed in some HIV patients have advanced the possibility that enforcement of self-tolerance may contribute to the rarity of BNAbs. To examine the possible breakdown of tolerance in HIV patients, we utilized the 9G4 anti-idiotype antibody system, enabling resolution of both autoreactive VH4-34 gene-expressing B cells and serum antibodies. Compared with healthy controls, HIV patients had significantly elevated 9G4+ serum IgG antibody concentrations and frequencies of 9G4+ B cells, a finding characteristic of systemic lupus erythematosus (SLE) patients, both of which positively correlated with HIV viral load. Compared to the global 9G4-IgD--memory B cell population, the 9G4+IgD--memory fraction in HIV patients was dominated by isotype switched IgG+ B cells, but had a more prominent bias toward "IgM only" memory. HIV envelope reactivity was observed both in the 9G4+ serum antibody and 9G4+ B cell population. 9G4+ IgG serum antibody levels positively correlated (r = 0.403, p = 0.0019) with the serum HIV BNAbs. Interestingly, other serum autoantibodies commonly found in SLE (anti-dsDNA, ANA, anti-CL) did not correlate with serum HIV BNAbs. 9G4-associated autoreactivity is preferentially expanded in chronic HIV infection as compared to other SLE autoreactivities. Therefore, the 9G4 system provides an effective tool to examine autoreactivity in HIV patients. Our results suggest that the development of HIV BNAbs is not merely a consequence of a general breakdown in tolerance, but rather
Comprehensive analysis of sperm DNA fragmentation by five different assays: TUNEL assay, SCSA, SCD test and alkaline and neutral Comet assay.
Ribas-Maynou, J; García-Peiró, A; Fernández-Encinas, A; Abad, C; Amengual, M J; Prada, E; Navarro, J; Benet, J
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.
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.
Wild, J; Schmiedel, B J; Maurer, A; Raab, S; Prokop, L; Stevanović, S; Dörfel, D; Schneider, P; Salih, H R
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.
Inertia-Centric Stability Analysis of a Planar Uniform Dust Molecular Cloud with Weak Neutral-Charged Dust Frictional Coupling
NASA Astrophysics Data System (ADS)
K. Karmakar, P.; Borah, B.
This paper adopts an inertia-centric evolutionary model to study the excitation mechanism of new gravito-electrostatic eigenmode structures in a one-dimensional (1-D) planar self-gravitating dust molecular cloud (DMC) on the Jeans scale. A quasi-neutral multi-fluid consisting of warm electrons, warm ions, neutral gas and identical inertial cold dust grains with partial ionization is considered. The grain-charge is assumed not to vary at the fluctuation evolution time scale. The neutral gas particles form the background, which is weakly coupled with the collapsing grainy plasma mass. The gravitational decoupling of the background neutral particles is justifiable for a higher inertial mass of the grains with higher neutral population density so that the Jeans mode frequency becomes reasonably large. Its physical basis is the Jeans assumption of a self-gravitating uniform medium adopted for fiducially analytical simplification by neglecting the zero-order field. So, the equilibrium is justifiably treated initially as "homogeneous". The efficacious inertial role of the thermal species amidst weak collisions of the neutral-charged grains is taken into account. A standard multiscale technique over the gravito-electrostatic equilibrium yields a unique pair of Korteweg-de Vries (KdV) equations. It is integrated numerically by the fourth-order Runge-Kutta method with multi-parameter variation for exact shape analyses. Interestingly, the model is conducive for the propagation of new conservative solitary spectral patterns. Their basic physics, parametric features and unique characteristics are discussed. The results go qualitatively in good correspondence with the earlier observations made by others. Tentative applications relevant to space and astrophysical environments are concisely highlighted.
MSFC Skylab neutral buoyancy simulator
NASA Technical Reports Server (NTRS)
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.
Comparative study of the cytolytic activity of snake venoms from African spitting cobras (Naja spp., Elapidae) and its neutralization by a polyspecific antivenom.
Méndez, Ileana; Gutiérrez, José María; Angulo, Yamileth; Calvete, Juan J; Lomonte, Bruno
Venoms of several Naja species found in Sub-Saharan Africa, and commonly known as "spitting cobras", induce a predominantly cytotoxic pattern of envenomings that may evolve into tissue necrosis and gangrene. Cytotoxic components of their venoms have been identified as members of the three-finger toxin and phospholipase A(2) protein families. In this study, an in vitro assay using the myogenic cell line C2C12, was utilized to compare the cytolytic activities of venoms from five species of spitting cobras: Naja nigricollis, Naja katiensis, Naja pallida, Naja nubiae, and Naja mossambica. These venoms were strongly cytotoxic, causing a 50% effect at ~1.5 μg/well (15 μg/ml), except for N. katiensis venom, which required nearly twice this amount. Using the cell-based assay, the ability of an equine polyspecific antivenom (EchiTab-Plus-ICP) to neutralize cytotoxicity was assessed. The antivenom completely inhibited the cytotoxic activity of all five venoms, although high antivenom/venom ratios were needed. Neutralization curves displayed the following decreasing order of efficiency: N. nubiae > N. pallida > N. mossambica > N. nigricollis > N. katiensis. Results indicate that neutralizing antibodies toward toxins responsible for this particular effect are present in the antivenom, albeit in low titers. Fucoidan, a natural sulfated polysaccharide known to inhibit the toxic effects of some basic snake venom components, was unable to reduce cytotoxicity of Naja venoms. Results emphasize the need of enhancing the immunogenicity of low molecular mass toxins during antivenom production, as well as to search for useful toxin inhibitors which could complement antivenom therapy.
Understanding low reliability of memories for neutral information encoded under stress: alterations in memory-related activation in the hippocampus and midbrain.
Qin, Shaozheng; Hermans, Erno J; van Marle, Hein J F; Fernández, Guillén
Exposure to an acute stressor can lead to unreliable remembrance of intrinsically neutral information, as exemplified by low reliability of eyewitness memories, which stands in contrast with enhanced memory for the stressful incident itself. Stress-sensitive neuromodulators (e.g., catecholamines) are believed to cause this low reliability by altering neurocognitive processes underlying memory formation. Using event-related functional magnetic resonance imaging, we investigated neural activity during memory formation in 44 young, healthy human participants while incidentally encoding emotionally neutral, complex scenes embedded in either a stressful or neutral context. We recorded event-related pupil dilation responses as an indirect index of phasic noradrenergic activity. Autonomic, endocrine, and psychological measures were acquired to validate stress manipulation. Acute stress during encoding led to a more liberal response bias (more hits and false alarms) when testing memory for the scenes 24 h later. The strength of this bias correlated negatively with pupil dilation responses and positively with stress-induced heart rate increases at encoding. Acute stress, moreover, reduced subsequent memory effects (SMEs; items later remembered vs forgotten) in hippocampus and midbrain, and in pupil dilation responses. The diminished SMEs indicate reduced selectivity and specificity in mnemonic processing during memory formation. This is in line with a model in which stress-induced catecholaminergic hyperactivation alters phasic neuromodulatory signaling in memory-related circuits, resulting in generalized (gist-based) processing at the cost of specificity. Thus, one may speculate that loss of specificity may yield less discrete memory representations at time of encoding, thereby causing a more liberal response bias when probing these memories.
An immunogen containing four tandem 10E8 epitope repeats with exposed key residues induces antibodies that neutralize HIV-1 and activates an ADCC reporter gene.
Sun, Zhiwu; Zhu, Yun; Wang, Qian; Ye, Ling; Dai, Yanyan; Su, Shan; Yu, Fei; Ying, Tianlei; Yang, Chinglai; Jiang, Shibo; Lu, Lu
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.
Prompt-Gamma Activation Analysis.
Lindstrom, Richard M
A permanent, 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 this 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.
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…
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...
Characterization of an invertase with pH tolerance and truncation of its N-terminal to shift optimum activity toward neutral pH.
Du, Liqin; Pang, Hao; Wang, Zilong; Lu, Jian; Wei, Yutuo; Huang, Ribo
Most invertases identified to date have optimal activity at acidic pH, and are intolerant to neutral or alkaline environments. Here, an acid invertase named uninv2 is described. Uninv2 contained 586 amino acids, with a 100 amino acids N-terminal domain, a catalytic domain and a C-terminal domain. With sucrose as the substrate, uninv2 activity was optimal at pH 4.5 and at 45°C. Removal of N-terminal domain of uninv2 has shifted the optimum pH to 6.0 while retaining its optimum temperaure at 45°C. Both uninv2 and the truncated enzyme retained highly stable at neutral pH at 37°C, and they were stable at their optimum pH at 4°C for as long as 30 days. These characteristics make them far superior to invertase from Saccharomyces cerevisiae, which is mostly used as industrial enzyme.
A constant threat for HIV: Fc-engineering to enhance broadly neutralizing antibody activity for immunotherapy of the acquired immunodeficiency syndrome.
Nimmerjahn, Falk
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.
Increased frequencies of CD8+CD57+ T cells are associated with antibody neutralization breadth against HIV in viraemic controllers
Palmer, Christine D; Romero-Tejeda, Marisol; Scully, Eileen P; Lockhart, Ainsley; Seaman, Michael S; Goldenthal, Ariel; Piechocka-Trocha, Alicja; Walker, Bruce D; Chibnik, Lori B; Jost, Stephanie; Porichis, Filippos
Introduction An effective prophylactic vaccine against HIV will need to elicit antibody responses capable of recognizing and neutralizing rapidly evolving antigenic regions. The immunologic milieu associated with development of neutralizing antibody breadth remains to be fully defined. In this study, we sought to identify immunological signatures associated with neutralization breadth in HIV controllers. We applied an immune monitoring approach to analyze markers of T cell and myeloid cell activation by flow cytometry, comparing broad neutralizers with low- and non-neutralizers using multivariate and univariate analyses. Methods Antibody neutralization breadth was determined, and cryopreserved peripheral blood mononuclear cells were stained for T cell and myeloid cell activation markers. Subjects were grouped according to neutralization breadth, and T cell and myeloid cell activation was analyzed by partial least squares discriminant analysis to determine immune signatures associated with high neutralization breadth. Results We show that neutralization breadth in HIV viraemic controllers (VC) was strongly associated with increased frequencies of CD8+CD57+ T cells and that this association was independent of viral load, CD4 count and time since HIV diagnosis. Conclusions Our data show elevated frequencies of CD8+CD57+ T cells in VC who develop neutralization breadth against HIV. This immune signature could serve as a potential biomarker of neutralization breadth and should be further investigated in other HIV-positive cohorts and in HIV vaccine trials. PMID:27938646
Computational Mechanistic Study of Redox-Neutral Rh(III)-Catalyzed C-H Activation Reactions of Arylnitrones with Alkynes: Role of Noncovalent Interactions in Controlling Selectivity.
Xing, Yang-Yang; Liu, Jian-Biao; Tian, Ying-Ying; Sun, Chuan-Zhi; Huang, Fang; Chen, De-Zhan
The mechanism of redox-neutral Rh(III)-catalyzed coupling reactions of arylnitrones with alkynes was investigated by density functional theory (DFT) calculations. The free energy profiles associated with the catalytic cycle, involving C(sp(2))-H activation, insertion of alkyne, transfer of O atom, cyclization and protodemetalation, are presented and analyzed. An overwhelming preference for alkyne insertion into Rh-C over Rh-O is observed among all pathways, and the most favorable route is determined. The pivalate-assisted C-H activation step is turnover-limiting, and the cyclization step determines the diastereoselectivity of the reaction, with the stereoselectivity arising mainly from the difference of noncovalent interactions in key transition states. The detailed mechanism of O atom transfer, Rh(III)-Rh(I)-Rh(III) versus Rh(III)-Rh(V)-Rh(III) cycle, is discussed.
Techniques of Neutralization and Persistent Sexual Abuse by Clergy: A Content Analysis of Priest Personnel Files From the Archdiocese of Milwaukee.
Spraitz, Jason D; Bowen, Kendra N
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.
Sequence analysis of cDNAs encoding the heavy and light chain variable regions of a WSSV-neutralizing monoclonal antibody.
Natividad, Karlo Dante T; Nomura, Nakao; Matsumura, Masatoshi
Antibodies raised against individual viral envelope proteins have been used in white spot syndrome virus (WSSV) neutralization assays. We report here the sequence analysis of cDNAs encoding the variable regions of a novel monoclonal antibody that binds to the viral envelope protein and neutralizes WSSV infection. The heavy and light variable chains are most homologous to VH7183 germline gene (AF120472) and IgVk RF germline gene (AJ235936), respectively. Database searches using the derived sequences predicted residues comprising CDR loops. The 12 amino acid residue of the heavy chain CDR3 is rich in negatively charged aspartic acid (25%) and did not show significant homology to any murine V gene available on the database. This study provides insights on the paratope-epitope interaction and can be used to identify compounds with comparable properties as the paratope leading to future development of drugs and vaccines for WSSV infection.
An analysis of post-vocalic /s-ʃ/ neutralization in Augsburg German: evidence for a gradient sound change
Bukmaier, Véronique; Harrington, Jonathan; Kleber, Felicitas
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
Metal-ion mutagenesis: conversion of a purple acid phosphatase from sweet potato to a neutral phosphatase with the formation of an unprecedented catalytically competent Mn(II)Mn(II) active site.
Mitić, Natasa; Noble, Christopher J; Gahan, Lawrence R; Hanson, Graeme R; Schenk, Gerhard
The currently accepted paradigm is that the purple acid phosphatases (PAPs) require a heterovalent, dinuclear metal-ion center for catalysis. It is believed that this is an essential feature for these enzymes in order for them to operate under acidic conditions. A PAP from sweet potato is unusual in that it appears to have a specific requirement for manganese, forming a unique Fe(III)-mu-(O)-Mn(II) center under catalytically optimal conditions (Schenk et al. Proc. Natl. Acad. Sci. U.S.A. 2005, 102, 273). Herein, we demonstrate, with detailed electron paramagnetic resonance (EPR) spectroscopic and kinetic studies, that in this enzyme the chromophoric Fe(III) can be replaced by Mn(II), forming a catalytically active, unprecedented antiferromagnetically coupled homodivalent Mn(II)-mu-(H)OH-mu-carboxylato-Mn(II) center in a PAP. However, although the enzyme is still active, it no longer functions as an acid phosphatase, having optimal activity at neutral pH. Thus, PAPs may have evolved from distantly related divalent dinuclear metallohydrolases that operate under pH neutral conditions by stabilization of a trivalent-divalent metal-ion core. The present Mn(II)-Mn(II) system models these distant relatives, and the results herein make a significant contribution to our understanding of the role of the chromophoric metal ion as an activator of the nucleophile. In addition, the detailed analysis of strain broadened EPR spectra from exchange-coupled dinuclear Mn(II)-Mn(II) centers described herein provides the basis for the full interpretation of the EPR spectra from other dinuclear Mn metalloenzymes.
Characterization of a plant-produced recombinant human secretory IgA with broad neutralizing activity against HIV
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
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
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.
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.
Food and drug cues activate similar brain regions: a meta-analysis of functional MRI studies.
Tang, D W; Fellows, L K; Small, D M; Dagher, A
In healthy individuals, food cues can trigger hunger and feeding behavior. Likewise, smoking cues can trigger craving and relapse in smokers. Brain imaging studies report that structures involved in appetitive behaviors and reward, notably the insula, striatum, amygdala and orbital frontal cortex, tend to be activated by both visual food and smoking cues. Here, by carrying out a meta-analysis of human neuro-imaging studies, we investigate the neural network activated by: 1) food versus neutral cues (14 studies, 142 foci) 2) smoking versus neutral cues (15 studies, 176 foci) 3) smoking versus neutral cues when correlated with craving scores (7 studies, 108 foci). PubMed was used to identify cue-reactivity imaging studies that compared brain response to visual food or smoking cues to neutral cues. Fourteen articles were identified for the food meta-analysis and fifteen articles were identified for the smoking meta-analysis. Six articles were identified for the smoking cue correlated with craving analysis. Meta-analyses were carried out using activation likelihood estimation. Food cues were associated with increased blood oxygen level dependent (BOLD) response in the left amygdala, bilateral insula, bilateral orbital frontal cortex, and striatum. Smoking cues were associated with increased BOLD signal in the same areas, with the exception of the insula. However, the smoking meta-analysis of brain maps correlating cue-reactivity with subjective craving did identify the insula, suggesting that insula activation is only found when craving levels are high. The brain areas identified here are involved in learning, memory and motivation, and their cue-induced activity is an index of the incentive salience of the cues. Using meta-analytic techniques to combine a series of studies, we found that food and smoking cues activate comparable brain networks. There is significant overlap in brain regions responding to conditioned cues associated with natural and drug rewards.
Direct on-line analysis of neutral analytes by dual sweeping via complexation and organic solvent field enhancement in nonionic MEKC.
Cao, Jun; Li, Bin; Chang, Yan-Xu; Li, Ping
Conventionally, neutral compounds cannot be separated by nonionic micelle capillary electrophoresis. In this report, the development of a novel on-line preconcentration technique combining dual sweeping based on complexation and organic solvent field enhancement is applied to the sensitive and selective analysis of three neutral glucosides: ginsenoside Rf, ginsenoside Rg1, and ginsenoside Re. Nonionic micelle detectability by CE is demonstrated through effective focusing of large sample volumes (up to 38% capillary length) using a dual sweeping mode. This results in a 50- to 130-fold improvement in the LODs relative to conventional injection method. Neutral compounds sweeping is examined in terms of analyte mobility dependence on borate complexation, solvent viscosity difference, and Brij-35 interaction. Enhanced focusing performance by this hyphenated method was demonstrated by a greater than fourfold reduction in glucoside bandwidth, as compared with common sweeping (devoid of organic solvent-mediated sweeping method in the sample matrices). Moreover, separation efficiencies greater than a million theoretical plates can be achieved by sweeping large sample volumes into narrow zones. The designated method was also tested for its ability to determine the presence of glucosides in the crude extracts obtained from plant sample.
Proposal of actively heated, long stem based Cs delivery system for diagnostic neutral beam source in ITER
NASA Astrophysics Data System (ADS)
Bansal, G.; Mishra, S.; Pandya, K.; Bandyopadhyay, M.; Soni, J.; Gahlaut, A.; Parmar, K. G.; Shah, S.; Phukan, A.; Roopesh, G.; Ahmed, I.; Chakraborty, A. K.; Singh, M. J.; Schunke, B.; Hemsworth, R.; Svensson, L.; Chareyre, J.; Graceffa, J.
Positioning of Cesium (Cs) oven modules in the complex interface dominated space envelope of a negative ion source such as Diagnostic Neutral Beam (DNB) source for ITER is a challenge not only for the designer of the ion source, but also that of remote handling. A more user friendly design of the Cs delivery could emerge from the consideration of a possibility of injecting the Cs from an oven located outside the vacuum envelope of the ion source, thereby ensuring an ease of Cs refilling and oven maintenance. The design of such a delivery system involves long transmission path of lengths ˜4 m, from ambient to vacuum. System design involves incorporation of a low loss transmission tube enveloped by highly reflective inner surface pipe to reduce the heat losses and therefore heating of the nearby systems. A combination of all metallic valves operated at high temperatures has been incorporated in such a way that the Cs refilling or oven maintenance can be done without breaking the ion source vacuum. Removable joints in the oven heating elements are provided at specific locations to remove the Cs oven for ion source maintenance. Experimental data on Cs transmission over such a long length, required for an effective design of a co-axial transmission, is not presently available. However, an experiment has been carried out in ITER-India making measurements of Cs distribution in coaxial transmission of a length of more than 5 m. These experiments incorporate an additional feature of multiple nozzle distributor based Cs delivery into the ion source which might help in reducing the need of multiple Cs ovens in large ion sources like ITER. The Cs flux from the oven is measured by surface ionization detector (SID). The angular distribution of the Cs flux is measured by a movable SID in linear direction and has been found in good agreement with the calculations. The Cs inventory in the oven reservoir was measured by electrical resistivity measurements methods. The paper
Redox regulation of neutral sphingomyelinase-1 activity in HEK293 cells through a GSH-dependent mechanism.
Martín, Sergio F; Sawai, Hirofumi; Villalba, José M; Hannun, Yusuf A
Phospholipases are essential enzymes in cellular signalling processes such as cellular differentiation, proliferation and apoptosis. Based on its high degree of homology with sequences of prokaryote SMases, a type of Mg(2+)-dependent PLC (nSMase-1) was recently discovered which displayed strong redox dependence for activity in vitro [F. Rodrigues-Lima, A.C. Fensome, M. Josephs, J. Evans, R.J. Veldman, M. Katan (2000), J. Biol. Chem. 275 (36) 28316-28325]. The aim of this work was to test the hypothesis that glutathione could be a natural regulator of nSMase-1 activity ex vivo. We studied how altering glutathione levels and redox ratio modulate nSMase-1 activity in a HEK293 cell line that ectopically overexpressed the nSMase-1 gene. Diminishing total glutathione with BSO without altering significantly the GSH/GSSG ratio did not affect nSMase-1 activity. Treatment of cells with diamide produced a transient decrease of total glutathione and a sharp, but also transient, decrease of the GSH/GSSG ratio. Under these conditions, nSMase-1 activity was temporarily activated and then returned to normal levels. Simultaneous treatment with BSO and diamide that resulted in permanent decreases of total glutathione and GSH/GSSG redox ratio produced a sustained activation of nSMase-1 activity. Taken together, these data indicate that altering the GSH/GSSG ratio by increasing GSSG or decreasing GSH levels, but not the total concentration of glutathione, modulates nSMase-1 activity. Our findings are the first evidence supporting the ex vivo regulation of nSMase-1 through a redox glutathione-dependent mechanism.
Apoptosis induced by capsaicin in prostate PC-3 cells involves ceramide accumulation, neutral sphingomyelinase, and JNK activation.
Sánchez, Ana Maria; Malagarie-Cazenave, Sophie; Olea, Nuria; Vara, Diana; Chiloeches, Antonio; Díaz-Laviada, Inés
Numerous studies have recently focused on the anticarcinogenic, antimutagenic, or chemopreventive activities of the main pungent component of red pepper, capsaicin (N-vanillyl-8-methyl-1-nonenamide). We have previously shown that, in the androgen-independent prostate cancer PC-3 cells, capsaicin inhibits cell growth and induces apoptosis through reactive oxygen species (ROS) generation [Apoptosis 11 (2006) 89-99]. In the present study, we investigated the signaling pathways involved in the antiproliferative effect of capsaicin. Here, we report that capsaicin apoptotic effect was mediated by ceramide generation which occurred by sphingomyelin hydrolysis. Using siRNA, we demonstrated that N-SMase expression is required for the effect of capsaicin on prostate cell viability. We then investigated the role of MAP kinase cascades, extracellular signal-regulated protein kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK, in the antiproliferative effect of capsaicin, and we confirmed that capsaicin could activate ERK and JNK but not p38 MAPK. Pharmacological inhibition of JNK kinase, as well as inhibition of ROS by the reducing agent N-acetylcysteine, prevented ceramide accumulation and capsaicin-induced cell death. However, inhibition of ceramide accumulation by the SMase inhibitor D609 did not modify JNK activation. These data reveal JNK as an upstream regulator of ceramide production. Capsaicin-promoted activation of ERK was prevented with all the inhibitors tested. We conclude that capsaicin induces apoptosis in PC-3 cells via ROS generation, JNK activation, ceramide accumulation, and second, ERK activation.
A Label as a Hidden Persuader: Chemists' Neutralization Concept.
ERIC Educational Resources Information Center
Schmidt, Hans-Jurgen
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…
Neutral buoyancy test evaluation of hardware and extravehicular activity procedures for on-orbit assembly of a 14 meter precision reflector
NASA Astrophysics Data System (ADS)
Heard, Walter L., Jr.; Lake, Mark S.
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.
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.
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.
Assessing SABU (Serum Anti Bisa Ular), the sole Indonesian antivenom: A proteomic analysis and neutralization efficacy study
Tan, Choo Hock; Liew, Jia Lee; Tan, Kae Yi; Tan, Nget Hong
Serum Anti Ular Bisa (SABU) is the only snake antivenom produced locally in Indonesia; however, its effectiveness has not been rigorously evaluated. This study aimed to assess the protein composition and neutralization efficacy of SABU. SDS polyacrylamide gel electrophoresis, size-exclusion liquid chromatography and shotgun proteomics revealed that SABU consists of F(ab')2 but a significant amount of dimers, protein aggregates and contaminant albumins. SABU moderately neutralized Calloselasma rhodostoma venom (potency of 12.7 mg venom neutralized per ml antivenom, or 121.8 mg venom per g antivenom protein) and Bungarus fasciatus venom (0.9 mg/ml; 8.5 mg/g) but it was weak against the venoms of Naja sputatrix (0.3 mg/ml; 2.9 mg/g), Naja sumatrana (0.2 mg/ml; 1.8 mg/g) and Bungarus candidus (0.1 mg/ml; 1.0 mg/g). In comparison, NPAV, the Thai Neuro Polyvalent Antivenom, outperformed SABU with greater potencies against the venoms of N. sputatrix (0.6 mg/ml; 8.3 mg/g), N. sumatrana (0.5 mg/ml; 7.1 mg/g) and B. candidus (1.7 mg/ml; 23.2 mg/g). The inferior efficacy of SABU implies that a large antivenom dose is required clinically for effective treatment. Besides, the antivenom contains numerous impurities e.g., albumins that greatly increase the risk of hypersensitivity. Together, the findings indicate that the production of SABU warrants further improvement. PMID:27869134
Assessing SABU (Serum Anti Bisa Ular), the sole Indonesian antivenom: A proteomic analysis and neutralization efficacy study.
Tan, Choo Hock; Liew, Jia Lee; Tan, Kae Yi; Tan, Nget Hong
Serum Anti Ular Bisa (SABU) is the only snake antivenom produced locally in Indonesia; however, its effectiveness has not been rigorously evaluated. This study aimed to assess the protein composition and neutralization efficacy of SABU. SDS polyacrylamide gel electrophoresis, size-exclusion liquid chromatography and shotgun proteomics revealed that SABU consists of F(ab')2 but a significant amount of dimers, protein aggregates and contaminant albumins. SABU moderately neutralized Calloselasma rhodostoma venom (potency of 12.7 mg venom neutralized per ml antivenom, or 121.8 mg venom per g antivenom protein) and Bungarus fasciatus venom (0.9 mg/ml; 8.5 mg/g) but it was weak against the venoms of Naja sputatrix (0.3 mg/ml; 2.9 mg/g), Naja sumatrana (0.2 mg/ml; 1.8 mg/g) and Bungarus candidus (0.1 mg/ml; 1.0 mg/g). In comparison, NPAV, the Thai Neuro Polyvalent Antivenom, outperformed SABU with greater potencies against the venoms of N. sputatrix (0.6 mg/ml; 8.3 mg/g), N. sumatrana (0.5 mg/ml; 7.1 mg/g) and B. candidus (1.7 mg/ml; 23.2 mg/g). The inferior efficacy of SABU implies that a large antivenom dose is required clinically for effective treatment. Besides, the antivenom contains numerous impurities e.g., albumins that greatly increase the risk of hypersensitivity. Together, the findings indicate that the production of SABU warrants further improvement.
Characterization of the variable regions of a chimpanzee monoclonal antibody with potent neutralizing activity against HIV-1.
Vijh-Warrier, S; Murphy, E; Yokoyama, I; Tilley, S A
The variable (V) regions of C108G, a potent neutralizing chimpanzee mAb against a glycan-dependent epitope in the V2 region of HIV-1 gp120, have been characterized for reactivity with human VH and VK family-specific antisera, and their nucleotide sequences have been determined and analysed. To our knowledge, this is the first study characterizing expressed chimpanzee VH and VK genes. Results show that C108G expresses members of the VH3 and VK1 families, the largest VH and VK families in humans, respectively. Nucleotide and amino acid sequence analyses reveal that C108G VH is most homologous to the human VH3 germline gene, hsigdp33 or V3-43, and the human JH4 minigene. The human germline VK1 gene that is most homologous to C108G VK, hsigk1012, was previously observed in unmutated form in a human autoantibody with anti-i red blood cell antigen specificity and in seven human Fabs and a mAb directed against epitopes overlapping the CD4-binding site of HIV-1 gp120. This germline gene was unmutated in three of the human Fabs and was somatically mutated in the other four Fabs and the mAb. In addition, the JK minigene was used in C108G VK, JK2, is apparently over-represented in anti-HIV-1 mAbs/Fabs; this minigene was used in 61% of the anti-gp120 human Fabs recently described and in three other anti-CD4-binding site human mAbs derived by EBV transformation. While the significance of these findings is unclear, they may suggest a bias in VK/JK gene usage and/or network regulation involving an hsigk1012/JK2 idiotope(s) in the antibody response to HIV-1. Both the C108G VH and VK genes showed evidence of somatic mutation and antigen selection that apparently occurred in vivo during chronic exposure to HIV-1 and its antigens. Surprisingly, this somatic mutation was most profound in the CDR3 region of C108G VK; this region shared only 48% nucleotide homology with hsigk1012 contrasted with a homology of 94% over the remainder of these two V gene sequences. Perhaps the most
In vivo selection for the enhancement of Thermotoga maritima exopolygalacturonase activity at neutral pH and low temperature.
Drone, Jullien; Dion, Michel; Tellier, Charles; Rabiller, Claude
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.
Neutral particle analyzer diagnostics on the TCV tokamak
Karpushov, Alexander N.; Duval, Basil P.; Schlatter, Christian; Afanasyev, Valery I.; Chernyshev, Fedor V.
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.
Neutral particle analyzer diagnostics on the TCV tokamak
NASA Astrophysics Data System (ADS)
Karpushov, Alexander N.; Duval, Basil P.; Schlatter, Christian; Afanasyev, Valery I.; Chernyshev, Fedor V.
Experimental apparatus and data analysis techniques used in neutral particle analyzer (NPA) diagnostics on the Tokamak à 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 0.6-8.0keV with a time resolution of 0.5-2.0ms 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 0.5-50keV energy range with a resolution of 60%-10% and a time resolution of 0.5-4.0ms. 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.
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
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…
Analysis of arginine and lysine methylation utilizing peptide separations at neutral pH and electron transfer dissociation mass spectrometry.
Snijders, Ambrosius P L; Hung, Ming-Lung; Wilson, Stuart A; Dickman, Mark J
Arginine and lysine methylation are widespread protein post-translational modifications. Peptides containing these modifications are difficult to retain using traditional reversed-phase liquid chromatography because they are intrinsically basic/hydrophilic and often fragment poorly during collision induced fragmentation (CID). Therefore, they are difficult to analyze using standard proteomic workflows. To overcome these caveats, we performed peptide separations at neutral pH, resulting in increased retention of the hydrophilic/basic methylated peptides before identification using MS/MS. Alternatively trifluoroacetic acid (TFA) was used for increased trapping of methylated peptides. Electron-transfer dissociation (ETD) mass spectrometry was then used to identify and characterize methylated residues. In contrast to previous reports utilizing ETD for arginine methylation, we observed significant amount of side-chain fragmentation. Using heavy methyl stable isotope labeling with amino acids in cell culture it was shown that, similar to CID, a loss of monomethylamine or dimethylamine from the arginine methylated side-chain during ETD can be used as a diagnostic to determine the type of arginine methylation. CID of lysine methylated peptides does not lead to significant neutral losses, but ETD is still beneficial because of the high charge states of such peptides. The developed LC MS/MS methods were successfully applied to tryptic digests of a number of methylated proteins, including splicing factor proline-glutamine-rich protein (SFPQ), RNA and export factor-binding protein 2 (REF2-I) and Sul7D, demonstrating significant advantages over traditional LC MS/MS approaches.
Fast targeted analysis of 132 acidic and neutral drugs and poisons in whole blood using LC-MS/MS.
Di Rago, Matthew; Saar, Eva; Rodda, Luke N; Turfus, Sophie; Kotsos, Alex; Gerostamoulos, Dimitri; Drummer, Olaf H
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.
Heavy Neutral Beam Probe for edge plasma analysis in Tokamaks. Annual progress report, December 1, 1992--November 30, 1993
Castracane, J.; Saravia, E.; Beckstead, J.; Aceto, S.
The contents of this report present the progress achieved to date on the Heavy Neutral Beam Probe project. This effort is an international collaboration in magnetic confinement fusion energy research sponsored by the US Department of Energy, Office of Energy Research (Confinement Systems Division) and the Centre Canadien de Fusion Magnetique (CCFM). The overall objective of the effort is to develop and apply a neutral particle beam to the study of edge plasma dynamics in discharges on the Tokamak de Varennes (TdeV) facility in Montreal, Canada. To achieve this goal, a research and development project was established to produce the necessary hardware to make such measurements and meet the scheduling requirements of the program. At present the project is in the middle of its second budget period with the instrumentation on-site at TdeV. The first half of this budget period was used to complete total system tests at InterScience, Inc., dismantle and ship the hardware to TdeV, re-assemble and install the HNBP on the tokamak. Integration of the diagnostic into the TdeV facility has progressed to the point of first beam production and measurement on the plasma. At this time, the HNBP system is undergoing final de-bugging prior to re-start of machine operation in early Fall of this year.
Directed evolution and mutagenesis of glutamate decarboxylase from Lactobacillus brevis Lb85 to broaden the range of its activity toward a near-neutral pH.
Shi, Feng; Xie, Yilong; Jiang, Junjun; Wang, Nannan; Li, Yongfu; Wang, Xiaoyuan
Glutamate decarboxylase (GAD) transforms l-glutamate into γ-aminobutyric acid (GABA) with the consumption of a proton. GAD derived from lactic acid bacteria exhibits optimum activity at pH 4.0-5.0 and significantly loses activity at near-neutral pH. To broaden the active range of the GAD GadB1 from Lactobacillus brevis Lb85 toward a near-neutral pH, irrational design using directed evolution and rational design using site-specific mutagenesis were performed. For directed evolution of GadB1, a sensitive high-throughput screening strategy based on a pH indicator was established. One improved mutant, GadB1(T17I/D294G/Q346H), was selected from 800 variants after one round of EP-PCR. It exhibited 3.9- and 25.0-fold increase in activity and catalytic efficiency, respectively at pH 6.0. Through site-specific mutagenesis, several improved mutants were obtained, with GadB1(E312S) being the best one. The combined mutant GadB1(T17I/D294G/E312S/Q346H) showed even higher catalytic efficiency, 13.1- and 43.2-fold that of wild-type GadB1 at pH 4.6 and 6.0, respectively. The amount of GABA produced in gadB1(T17I/D294G/Q346H)-, gadB1(E312S)- and gadB1(T17I/D294G/E312S/Q346H)-expressing Corynebacterium glutamicum ATCC 13032 from endogenous l-glutamate increased by 9.6%, 20.3% and 63.9%, respectively. These results indicate that these mutations have beneficial effects on expanding the active pH range and on GABA biosynthesis, suggesting these GadB1 variants as potent candidates for GABA production.
Generation and reactivity of putative support systems, Ce-Al neutral binary oxide nanoclusters: CO oxidation and C-H bond activation
NASA Astrophysics Data System (ADS)
Wang, Zhe-Chen; Yin, Shi; Bernstein, Elliot R.
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.
The chimeric antibody chLpMab-7 targeting human podoplanin suppresses pulmonary metastasis via ADCC and CDC rather than via its neutralizing activity
Ogasawara, Satoshi; Fujii, Yuki; Oki, Hiroharu; Fukayama, Masashi; Nishioka, Yasuhiko; Kaneko, Mika K.
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
Neutralizing properties of Musa paradisiaca L. (Musaceae) juice on phospholipase A2, myotoxic, hemorrhagic and lethal activities of crotalidae venoms.
Borges, M H; Alves, D L F; Raslan, D S; Piló-Veloso, D; Rodrigues, V M; Homsi-Brandeburgo, M I; de Lima, M E
The use of plants as medicine has been referred to since ancient peoples, perhaps as early as Neanderthal man. Plants are a source of many biologically active products and nowadays they are of great interest to the pharmaceutical industry. The study of how people of different culture use plants in particular ways has led to the discovery of important new medicines. In this work, we verify the possible activity of Musa paradisiaca L. (Musaceae) against the toxicity of snake venoms. Musa paradisiaca, an important source of food in the world, has also been reported to be popularly used as an anti-venom. Interaction of Musa paradisiaca extract (MsE) with snake venom proteins has been examined in this study. Phospholipase A2 (PLA2), myotoxic and hemorrhagic activities, including lethality in mice, induced by crotalidae venoms were significantly inhibited when different amounts of MsE were mixed with these venoms before assays. On the other hand, mice that received MsE and venoms without previous mixture or by separated routes were not protected against venom toxicity. Partial chemical characterization of MsE showed the presence of polyphenols and tannins and they are known to non-specifically inactivate proteins. We suggest that these compounds can be responsible for the in vitro inhibition of the toxic effects of snake venoms. In conclusion, according to our results, using mice as experimental model, MsE does not show protection against the toxic effects of snake venoms in vivo, but if was very effective when the experiments were done in vitro.
Potent neutralization of VEGF biological activities with a fully human antibody Fab fragment directed against VEGF receptor 2
Miao, H.-Q. . E-mail: email@example.com; Hu, Kun; Jimenez, Xenia; Navarro, Elizabeth; Zhang, Haifan; Lu Dan; Ludwig, Dale L.; Balderes, Paul; Zhu Zhenping . E-mail: firstname.lastname@example.com
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.
CDAW 7 revisited - Further evidence for the creation of a near-earth substorm neutral line. [Coordinated Data Analysis Workshop
NASA Technical Reports Server (NTRS)
Kettmann, G.; Fritz, T. A.; Hones, E. W., Jr.
Eastman et al. (1988) have interpreted the CDAW 7 substorm of April 24, 1979, previously taken as unambiguously supporting the near-earth neutral line model of magnetospheric substorms, in terms of spatial movements of a preexisting plasma-sheet boundary layer (PSBL) and its associated current sheets across the observing ISEE 1 and 2 spacercraft. It is presently noted that, by contrast, a reinvestigation of ISEE 1 and 2 energetic particle measurements around substorm onset on short time-scales shows the observed flux pattern to require the formation of a particle source eastward of the ISEE spacecraft, well within the plasma sheet, associated with the substorm onset. Strong flows were absent prior to substorm onset, indicating the temporal nature of the event, as opposed to an encounter with a preexisting PSBL containing large flows.
Geographical variability of the venoms of four populations of Bothrops asper from Panama: Toxicological analysis and neutralization by a polyvalent antivenom.
Vélez, Sara María; Salazar, Marcos; Acosta de Patiño, Hildaura; Gómez, Leandra; Rodriguez, Abdiel; Correa, David; Saldaña, Julio; Navarro, Deyvi; Lomonte, Bruno; Otero-Patiño, Rafael; Gutiérrez, José María
Bothrops asper is the medically most important venomous snake in Central America. In Panama, the country having the highest incidence of snakebites in Latin America, B. asper is widely distributed throughout the country and is responsible for the vast majority of snakebites. This study was performed to analyze whether there are variations in the toxicological profile and in some biochemical parameters between the venoms of B. asper from four different regions in Panama. The venoms showed a similar profile of lethal, hemorrhagic, in vitro coagulant, defibrinogenating, edema-forming, myotoxic and indirect hemolytic activities, with subtle quantitative variations between samples of some regions. The venoms also had similar SDS-PAGE patterns and reverse phase HPLC profiles. A polyvalent antivenom manufactured in Costa Rica, and regularly used in Panama, was effective in the neutralization of lethal activity of the venoms of the four populations, with Mean Effective Doses (ED50) ranging from 5.98 to 9.72 mg venom/mL antivenom. In agreement, a widespread pattern of cross-reactivity between this antivenom and the four venoms was observed by immunoblotting. Overall, results highlight the lack of marked differences between the venoms of the various populations of B. asper in Panama, and that the antivenom from Costa Rica is effective in neutralizing lethality.
An in-source multiple collision-neutral loss filtering based nontargeted metabolomics approach for the comprehensive analysis of malonyl-ginsenosides from Panax ginseng, P. quinquefolius, and P. notoginseng.
Shi, Xiao-Jian; Yang, Wen-Zhi; Qiu, Shi; Yao, Chang-Liang; Shen, Yao; Pan, Hui-Qin; Bi, Qi-Rui; Yang, Min; Wu, Wan-Ying; Guo, De-An
The simultaneous identification and quantification of target metabolites from herbal medicines are difficult to implement by the full-scan MS based nontargeted metabolomics approaches. Here an in-source multiple collision-neutral loss filtering (IMC-NLF) based nontargeted metabolomics approach is developed and applied to identify and quantify the variations of malonyl-ginsenosides, a common group of acyl saponins with potential anti-diabetic activity, among Panax ginseng, P. quinquefolius, and P. notoginseng. The key steps of the IMC-NLF strategy are the acquisition of specific high-resolution neutral loss data and the efficient filtering of target precursor ions from the full-scan spectra. Using a hybrid LTQ-Orbitrap mass spectrometer after UHPLC separation, abundant in-source product ions, [M-H-CO2](-) (due to the vulnerability of the carboxyl group) and [M-H-Mal.](-), were generated at the energies of 70 V and 90 V, respectively. After spectral deconvolution, the generated peak list was screened by dual NLF using a Neutral Loss MS Finder software (NL of 43.9898 Da for CO2 and 86.0004 Da for the malonyl substituent). By combining the precursor ions list-triggered HCD-MS/MS and basic hydrolysis, a total of 101 malonyl-ginsenosides (including 69 from P. ginseng, 52 from P. quinquefolius, and 44 from P. notoginseng) were identified or tentatively characterized. The variations of 81 characterized malonyl-ginsenosides among 45 batches of Ginseng samples were statistically analyzed disclosing ten potential markers. It is the first systematic analysis of malonyl-ginsenosides. The IMC-NLF approach by a single analytical platform is promising in targeted analyses of modification-specific metabolites in metabolomics and drug metabolism.
Selective neutrality and enzyme kinetics.
Demetrius, L
This article appeals to a recent theory of enzyme evolution to show that the properties, neutral or adaptive, which characterize the observed allelic variation in natural populations can be inferred from the functional parameters, substrate specificity, and reaction rate. This study delineates the following relations between activity variables, and the forces--adaptive or neutral--determining allelic variation: (1) Enzymes with broad substrate specificity: The observed polymorphism is adaptive; mutations in this class of enzymes can result in increased fitness of the organism and hence be relevant for positive selection. (2) Enzymes with absolute substrate specificity and diffusion-controlled rates: Observed allelic variation will be absolutely neutral; mutations in this class of enzymes will be either deleterious or have no effect on fitness. (3) Enzymes with absolute or group specificity and nondiffusion-controlled rates: Observed variation will be partially neutral; mutants which are selectively neutral may become advantageous under an appropriate environmental condition or different genetic background. We illustrate each of the relations between kinetic properties and evolutionary states with examples drawn from enzymes whose evolutionary dynamics have been intensively studied.
Energetic Neutral Atom Precipitation (ENAP)
NASA Technical Reports Server (NTRS)
Tinsley, B. A.
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.
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.
Murali, Malliga Raman; Raja, Subramaniya Bharathi; Devaraj, Sivasitambaram Niranjali
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.
Canine distemper virus neutralization activity is low in human serum and it is sensitive to an amino acid substitution in the hemagglutinin protein.
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
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.
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.
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.
Pesticide residue analysis of a dietary ingredient by gas chromatography/selected-ion monitoring mass spectrometry using neutral alumina solid-phase extraction cleanup.
Jeong, Mijeong Lee; Zahn, Michael; Trinh, Thao; Brooke, Fay A; Ma, Wenwen
A sample cleanup procedure has been developed to remove coextractives that interfere with pesticide residue analysis of a dietary ingredient (Product B), an extract consisting of Scutellaria baicalensis and Acacia catechu. Samples were extracted using 1% acetic acid in acetonitrile, followed by solid-phase extraction and analysis by capillary gas chromatography with mass spectrometry in the selective-ion monitoring mode. Neutral alumina (alumina N) was found to be the most effective sorbent to remove coextractives from Product B; other materials that were tested but failed to remove interference were graphitized carbon black/primary-secondary amine (PSA), octadecylsilane (C18), Florisil, Oasis MCX, and strong anion exchange-PSA. The method was specifically developed for Product B, which was spiked with 41 organochlorine and organophosphorus pesticides, and resulted in the recovery of 80 to 120% at U.S. Pharmacopeia limits (0.06 to 4 microg/g) for the majority of the pesticides.
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.
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
Effects of Physically Effective Neutral Detergent Fiber Content on Intake, Digestibility, and Chewing Activity in Fattening Heifer Fed Total Mixed Ration
Oh, Mi Rae; Hong, Heeok; Li, Hong Liang; Jeon, Byong Tae; Choi, Cheong Hee; Ding, Yu Ling; Tang, Yu Jiao; Kim, Eun Kyung; Jang, Se Young; Seong, Hye Jin; Moon, Sang Ho
The objective of this study was to determine the effects of physically effective neutral detergent fiber (peNDF) content in total mixed ration (TMR) on dry matter intake, digestibility, and chewing activity in fattening Hanwoo (Bos taurus coreanae) heifers. The experiment was designed as a replicated 3×3 Latin square using 12 heifers. Fattening heifers were offered one of three diets [high (T1), medium (T2), and low (T3) peNDF] obtained by different mixing times (3, 10, and 25 min) for the same TMR feed. The peNDF content of TMR was determined by multiplying the proportion of dry matter retained by a 1.18 mm-screen in a Penn State Particle Separator by the dietary NDF content. The peNDF1.18 content was 30.36%, 29.20%, and 27.50% for the T1, T2, and T3 diets, respectively (p<0.05). Dry matter intake was not affected by peNDF content in TMR. Total weight gain in T1 group was significantly higher (p<0.05) than in T2 and T3 groups. However, weight gain did not differ between T2 and T3 groups. The feed conversion ratio decreased with an increase in the peNDF content (T1: 12.18, T2: 14.17, and T3: 14.01 g/g). An increase in the peNDF content of TMR was associated with a linear increase in the digestibility of dry matter, crude protein, crude fiber, neutral detergent fiber, and acid detergent fiber (p<0.05). Also, an increase in peNDF content of the TMR resulted in a linear increase in the number of chews in eating and ruminating (p<0.05), and consequently in the number of total chews (p<0.05). These results indicate that peNDF content affects digestibility and chewing activity. Consequently, the peNDF content of TMR should be considered for improving feed efficiency, digestibility, body weight gain, and performance in fattening heifers. PMID:27488845
Total body nitrogen analysis. [neutron activation analysis
NASA Technical Reports Server (NTRS)
Palmer, H. E.
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.
Phylogenetic analysis of the main neutralization and hemagglutination determinants of all human adenovirus prototypes as a basis for molecular classification and taxonomy.
Madisch, Ijad; Harste, Gabi; Pommer, Heidi; Heim, Albert
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.
Neutral-current detectors for the Sudbury Neutrino Observatory
Hime, A.; SNO Collaboration
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.
Neutral Higgs boson production at e^+e^- colliders in the complex MSSM: a full one-loop analysis
NASA Astrophysics Data System (ADS)
Heinemeyer, S.; Schappacher, C.
For the search for additional Higgs bosons in the Minimal Supersymmetric Standard Model (MSSM) as well as for future precision analyses in the Higgs sector precise knowledge of their production properties is mandatory. We evaluate the cross sections for the neutral Higgs boson production at e^+e^- colliders in the MSSM with complex parameters (cMSSM). The evaluation is based on a full one-loop calculation of the production mechanism e^+e^- → h_i Z, h_i γ , h_i h_j (i,j = 1,2,3), including soft and hard QED radiation. The dependence of the Higgs boson production cross sections on the relevant cMSSM parameters is analyzed numerically. We find sizable contributions to many cross sections. They are, depending on the production channel, roughly of 10-20 % of the tree-level results, but can go up to 50 % or higher. The full one-loop contributions are important for a future linear e^+e^- collider such as the ILC or CLIC. There are plans to implement the evaluation of the Higgs boson production cross sections into the code FeynHiggs.
Analysis of the Henderson-Abraham-Barker equation in the case of a polar liquid near a neutral hard wall
NASA Astrophysics Data System (ADS)
Badiali, J. P.; Russier, V.; Holovko, M. E.
The physical content of the wall-particle direct correlation function cw(1), defined from the Ornstein-Zernike equation in the framework of the Henderson-Abraham-Barker approach, is analyzed in the case of a dipolar-hard-sphere fluid near a pure and dielectric neutral wall. The exact asymptotic behavior of cw(1) is established and we show that it is not related to simple physical concepts as, e.g., the image potential. We show that the exact Henderson-Abraham-Barker equation introduces some bridge diagrams which are more simple graphs in another approach. Due to this fact, at least for systems with long range interactions, it is misleading to use the usual closures of the theory of homogeneous liquids for cw(1). In the case of a dielectric wall, we emphasize that the diagrammatic structure of cw(1) requires the introduction of a three-body Mayer function. The dipolar-hard-sphere liquid is a good candidate for analyzing cw(1) because some exact results are known and related to simple electrostatic effects; however, the present results are not restricted to this system.
Analysis of γ-ray production in neutral-current neutrino-oxygen interactions at energies above 200 MeV.
Ankowski, Artur M; Benhar, Omar; Mori, Takaaki; Yamaguchi, Ryuta; Sakuda, Makoto
It has long been recognized that the observation of γ rays originating from nuclear deexcitation can be exploited to identify neutral-current neutrino-nucleus interactions in water-Cherenkov detectors. We report the results of a calculation of the neutrino- and antineutrino-induced γ-ray production cross section for the oxygen target. Our analysis is focused on the kinematical region of neutrino energy larger than ∼200 MeV, in which a single-nucleon knockout is known to be the dominant reaction mechanism. The numerical results have been obtained using for the first time a realistic model of the target spectral function, extensively tested against electron-nucleus scattering data. We find that at a neutrino energy of 600 MeV the fraction of neutral-current interactions leading to emission of γ rays of energy larger than 6 MeV is ∼41%, and that the contribution of the p_{3/2} state is overwhelming.
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.
Kumar, Vidya P; Basavannacharya, Chandrakala; de Sousa, Sunita M
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.
Characterization and epitope mapping of monoclonal antibodies raised against rat hepatitis E virus capsid protein: An evaluation of their neutralizing activity in a cell culture system.
Kobayashi, Tominari; Takahashi, Masaharu; Tanggis; Mulyanto; Jirintai, Suljid; Nagashima, Shigeo; Nishizawa, Tsutomu; Okamoto, Hiroaki
Hepatitis E virus (HEV) is the causative agent of acute hepatitis. Rat HEV is a recently discovered virus related to, but distinct from, human HEV. Since laboratory rats can be reproducibly infected with rat HEV and a cell culture system has been established for rat HEV, this virus may be used as a surrogate virus for human HEV, enabling studies on virus replication and mechanism of infection. However, monoclonal antibodies (MAbs) against rat HEV capsid (ORF2) protein are not available. In this study, 12 murine MAbs were generated against a recombinant ORF2 protein of rat HEV (rRatHEV-ORF2: amino acids 101-644) and were classified into at least six distinct groups by epitope mapping and a cross-reactivity analysis with human HEV ORF2 proteins. Two non-cross-reactive MAbs recognizing the protruding (P) domain detected both non-denatured and denatured rRatHEV-ORF2 protein and efficiently captured cell culture-produced rat HEV particles that had been treated with deoxycholate and trypsin, but not those without prior treatment. In addition, these two MAbs were able to efficiently neutralize replication of cell culture-generated rat HEV particles without lipid membranes (but not those with lipid membranes) in a cell culture system, similar to human HEV.
In vitro assay for neutralizing antibody to hepatitis C virus: evidence for broadly conserved neutralization epitopes.
Bartosch, Birke; Bukh, Jens; Meunier, Jean-Christophe; Granier, Christelle; Engle, Ronald E; Blackwelder, William C; Emerson, Suzanne U; Cosset, François-Loïc; Purcell, Robert H
Our understanding of the humoral immune response to hepatitis C virus (HCV) is limited because the virus can be studied only in humans and chimpanzees and because previously described neutralization assays have not been robust or simple to perform. Nevertheless, epidemiologic and laboratory studies suggested that neutralizing Ab to HCV might be important in preventing infection. We have recently described a neutralization assay based on the neutralization of pseudotyped murine retrovirus constructs bearing HCV envelope glycoproteins on their surface. We have applied the assay to well characterized clinical samples from HCV-infected patients and chimpanzees, confirmed the existence of neutralizing Ab to HCV, and validated most previously reported neutralizations of the virus. We did not find neutralizing anti-HCV in resolving infections but did find relatively high titers (>1:320) of such Ab in chronic infections. Neutralizing Ab was directed not only to epitope(s) in the hypervariable region of the E2 envelope protein but also to one or more epitopes elsewhere in the envelope of the virus. Neutralizing Ab was broadly reactive and could neutralize pseudotype particles bearing the envelope glycoproteins of two different subgenotypes (1a and 1b). The ability to assay neutralizing anti-HCV should permit an assessment of the prospects for successful Ab-mediated passive and active immunoprophylaxis against hepatitis C.
In vitro assay for neutralizing antibody to hepatitis C virus: Evidence for broadly conserved neutralization epitopes
Bartosch, Birke; Bukh, Jens; Meunier, Jean-Christophe; Granier, Christelle; Engle, Ronald E.; Blackwelder, William C.; Emerson, Suzanne U.; Cosset, François-Loïc; Purcell, Robert H.
Our understanding of the humoral immune response to hepatitis C virus (HCV) is limited because the virus can be studied only in humans and chimpanzees and because previously described neutralization assays have not been robust or simple to perform. Nevertheless, epidemiologic and laboratory studies suggested that neutralizing Ab to HCV might be important in preventing infection. We have recently described a neutralization assay based on the neutralization of pseudotyped murine retrovirus constructs bearing HCV envelope glycoproteins on their surface. We have applied the assay to well characterized clinical samples from HCV-infected patients and chimpanzees, confirmed the existence of neutralizing Ab to HCV, and validated most previously reported neutralizations of the virus. We did not find neutralizing anti-HCV in resolving infections but did find relatively high titers (>1:320) of such Ab in chronic infections. Neutralizing Ab was directed not only to epitope(s) in the hypervariable region of the E2 envelope protein but also to one or more epitopes elsewhere in the envelope of the virus. Neutralizing Ab was broadly reactive and could neutralize pseudotype particles bearing the envelope glycoproteins of two different subgenotypes (1a and 1b). The ability to assay neutralizing anti-HCV should permit an assessment of the prospects for successful Ab-mediated passive and active immunoprophylaxis against hepatitis C. PMID:14617769
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
Brents, Lisa K.; Gallus-Zawada, Anna; Radominska-Pandya, Anna; Vasiljevik, Tamara; Prisinzano, Thomas E.; Fantegrossi, William E.; Moran, Jeffery H.; Prather, Paul L.
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
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.
Brents, Lisa K; Gallus-Zawada, Anna; Radominska-Pandya, Anna; Vasiljevik, Tamara; Prisinzano, Thomas E; Fantegrossi, William E; Moran, Jeffery H; Prather, Paul L
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.
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
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.
Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle: Mechanistic insights into outer-membrane permeabilization and endotoxin neutralization.
Saravanan, Rathi; Mohanram, Harini; Joshi, Mangesh; Domadia, Prerna N; Torres, Jaume; Ruedl, Christiane; Bhattacharjya, Surajit
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.
The effects of adding fat to diets of lactating dairy cows on total-tract neutral detergent fiber digestibility: A meta-analysis.
Weld, K A; Armentano, L E
The objective of this meta-analysis was to determine the effects of supplemental fat on fiber digestibility in lactating dairy cattle. Published papers that evaluated the effects of adding fat to the diets of lactating dairy cattle on total-tract neutral detergent fiber digestibility (ttNDFd) and dry matter intake (DMI) were compiled. The final data set included 108 fat-supplemented treatment means, not including low-fat controls, from 38 publications. The fat-supplemented treatment means exhibited a wide range of ttNDFd (49.4% ± 9.3, mean ± standard deviation) and DMI (21.3 kg/d ± 3.5). Observations were summarized as the difference between the treatment means for fat-supplemented diets minus their respective low-fat control means. Additionally, those differences were divided by the difference in diet fatty acid (FA) concentration between the treatment and control diets. Treatment means were categorized by the type of fat supplement. Supplementing 3% FA in the diet as medium-chain fats (containing predominately 12- and 14-carbon saturated FA) or unsaturated vegetable oil decreased ttNDFd by 8.0 and 1.2 percentage units, respectively. Adding 3% calcium salts of long-chain FA or saturated fats increased ttNDFd by 3.2 and 1.3 percentage units, respectively. No other fat supplement type affected ttNDFd. Except for saturated fats and animal-vegetable fats, supplementing dietary fat decreased DMI. When the values for changes in ttNDFd are regressed on changes in DMI there was a positive relationship, though the coefficient of determination is only 0.20. When changes in ttNDFd were regressed on changes in DMI, within individual fat supplement types, there was no relationship within calcium salt supplements. There was a positive relationship between changes in ttNDFd and changes in DMI for saturated fats. Neither relationship suggested that the increased ttNDFd with calcium salts or saturated FA was due to decreased DMI for these fat sources. A subset of the means
Origins and variation of terrestrial energetic neutral atoms outflow
NASA Astrophysics Data System (ADS)
Wilson, G. R.; Moore, T. E.
Analysis of ENA data from the LENA instrument on the IMAGE spacecraft shows that the terrestrial atmosphere is a copious emitter of energetic neutral atoms (<300 eV) under all conditions. When activity is low, the observed emissions are concentrated close to the Earth and are presumed to be the high-energy tail of the warm oxygen geocorona, with energies <2 eV. When activity increases, the relative abundance of the higher-energy neutrals increases, and the emissions can be seen farther from the Earth. Because of the close correlation between the postperigee ENA flux (fluxes seen 1-2 hours after spacecraft perigee) and Ap and the fact that the postperigee fluxes are seen when no magnetic storm is in progress we conclude that many of the emitted ENA come from the auroral zone and are produced by energized ionospheric ions rather than by precipitating energetic ions. In more spectacular events, such as the Bastille Day storm event (14-16 July 2000), oxygen neutral emissions produced by precipitation of keV ring current oxygen ions can also make an important contribution to the total neutral emission. We conclude that diurnal variation in ENA emissions is a winter hemisphere feature that is absent in the summer hemisphere. As activity increases, the altitude range of the auroral oval ENA emission region increases.
Neutron Activation Analysis of Water - A Review
NASA Technical Reports Server (NTRS)
Buchanan, John D.
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.
NASA Technical Reports Server (NTRS)
Liu, W. Timothy; Tang, Wenqing
The definition of equivalent neutral wind and the rationale for using it as the geophysical product of a spaceborne scatterometer are reviewed. The differences between equivalent neutral wind and actual wind, which are caused by atmospheric density stratification, are demonstrated with measurements at selected locations. A method of computing this parameter from ship and buoy measurements is described and some common fallacies in accounting for the effects of atmospheric stratification on wind shear are discussed. The computer code for the model to derive equivalent neutral wind is provided.
A new EV71 VP3 epitope in norovirus P particle vector displays neutralizing activity and protection in vivo in mice.
Jiang, Liping; Fan, Rongjun; Sun, Shiyang; Fan, Peihu; Su, Weiheng; Zhou, Yan; Gao, Feng; Xu, Fei; Kong, Wei; Jiang, Chunlai
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.
NASA Astrophysics Data System (ADS)
Ghosh, S. N.
After World War II, Professor S.K. Mitra wrote a comprehensive book called The Upper Atmosphere, which dealt with information available from ground-based and balloon-borne experiments. As a result, topics such as day airglow were investigated and further ground-based experiments using incoherent back-scattering were carried out. These activities resulted in important new information on the ozonosphere. The dramatic discovery of ozone holes forms a new and exciting chapter in the discovery of atmospheric processes. While dealing with the limits of the atmosphere, reference may be made to interstellar molecules whose discovery has raised considerable scientific curiosity. Knowledge on the solar-terrestrial relationship advanced a great deal when more information on solar radiation became available by measuring higher energy photons in the UV, EUV, and even X-ray regime. All this information is incorporated in this volume and presented under the title The Neutral Upper Atmosphere. Link: http://www.wkap.nl/prod/b/0-7923-6434-1
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 ...
NASA Astrophysics Data System (ADS)
Poncet, A.
After a short presentation of intensity limitations examples due to trapped ions, the processes of ionization and neutralization build up in particle accelerators and storage rings are briefly reviewed. The tolerable limits in neutralization are then assessed at the light of current theories of incoherent and coherent effects driven by ions. Finally the usual antidotes such as clearing electrodes, missing bunch schemes and beam shaking are presented.
Analysis of plasma and neutral gas flow inside of a PET bottle under PIII condition by particle-in-cell/Monte Carlo simulation
NASA Astrophysics Data System (ADS)
Miyagawa, Y.; Tanaka, M.; Ikeyama, M.; Nakao, S.; Choi, J.; Miyagawa, S.
The plasma behavior inside of a PET bottle has been simulated under the condition of plasma immersed ion implantation and deposition (PIII&D) using the simulation software "PEGASUS". The software uses the "PIC-MCCM" module for the plasma analysis and the "DSMCM" module for the gas flow field analysis. DSMCM gives densities, velocities, fluxes, temperatures and pressures of each neutral species such as the fed gas species and radicals. By coupling PIC-MCCM with DSMCM simulation, the plasma behavior in the flowing Ar gas and N2 gas has been simulated. The gas was injected from the tip of the gas inlet which was inserted into the center of the bottle. The base gas pressure was 1-50 Pa and a positive pulse voltage (maximum voltage = 0.1-1 kV) was applied to the center rod. A two-dimensional cylindrical coordinate system was used. Time evolution of the spacial distribution was obtained for densities of electrons, N2+ ions, N2∗ radicals and N atoms in N2 gas, and Ar+ ions, Ar∗ and Ar∗(4s) radicals in Ar gas. Time evolution of the particle flux and the energy flux of electrons and ions on the target surface was also obtained.
Characterizing and compensating for matrix effects using atmospheric pressure chemical ionization liquid chromatography-tandem mass spectrometry: analysis of neutral pharmaceuticals in municipal wastewater.
Zhao, Xiaoming; Metcalfe, Chris D
Matrix effects are a great challenge for the quantitative analysis of environmental samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Signal suppression or enhancement can compromise the accuracy of analytical results. While matrix effects have been relatively well studied for applications of LC-MS/MS instrumentation with electrospray ionization, there have been relatively few studies to evaluate matrix effects when using atmospheric pressure chemical ionization (APCI) as the ion source. In this study, we determined the effects of sample matrix on the analysis of six neutral pharmaceuticals (i.e., caffeine, cotinine, carbamazepine and its major metabolite, carbamazepine-10,11-dihydrodiol, trimethoprim, and fluoxetine) in samples of municipal wastewater using LC-APCI-MS/MS and evaluated whether isotope-labeled internal standards can be used to compensate for matrix effects. The matrix effects were measured using postextraction spikes and postcolumn direct infusion, respectively. The results showed that the matrix in the extracts prepared from municipal wastewater enhanced the signals for four of the six analytes when using an APCI source. Without correction for signal enhancement, apparent recoveries of the analytes from wastewater samples were overestimated to levels as high as 178% of the spiked amount. Isotope-labeled compounds corrected for these overestimates that occurred as a result of interferences from the sample matrix.
Serial mixed-mode cation- and anion-exchange solid-phase extraction for separation of basic, neutral and acidic pharmaceuticals in wastewater and analysis by high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry.
Lavén, Martin; Alsberg, Tomas; Yu, Yong; Adolfsson-Erici, Margaretha; Sun, Hongwen
A novel solid-phase extraction (SPE) method is presented whereby 15 basic, neutral and acidic pharmaceuticals in wastewater were simultaneously extracted and subsequently separated into different fractions. This was achieved using mixed-mode cation- and anion-exchange SPE (Oasis MCX and MAX) in series. Analysis was performed by high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC/QTOF-MS). A fast separation was achieved, with all compounds eluting within 6min, narrow chromatographic peaks, with a peak base width of 6s on average, and a high mass accuracy of quantified wastewater sample ions, with average mass errors in absolute value of 0.7mDa or 2.7ppm. The recovery of the SPE method in the analysis of sewage treatment plant (STP) influent and effluent wastewater was on average 80% and the ion suppression 30%. For less demanding samples Oasis MCX used alone may be an alternative method, although for STP influent waters containing high loads of organic compounds the clean-up achieved using only Oasis MCX was insufficient, leading to unreliable quantitation. Furthermore, serial SPE separation according to molecular charge added an additional degree of analyte confirmation. For quantitation, an approach combining external standard calibration curves, isotopically labelled surrogate standards and single-point standard addition was used. The applicability of the method was demonstrated in the analysis of influent and effluent wastewater from an STP, using small sample volumes (25-50mL). The effluent wastewater had been subjected to three different treatments; activated sludge, activated sludge followed by ozonation, and a membrane bioreactor (MBR). Ozone treatment proved superior in removal of the analysed pharmaceuticals, while the MBR provided higher removal efficiencies than the activated sludge process.
Neutral matter in planetary nebulae
NASA Technical Reports Server (NTRS)
Dinerstein, Harriet L.
A review of current studies of neutral envelopes is presented with particular attention given to the use of the envelopes as test cases for understanding the ionization and thermal structure of photodissociation regions. The study of near-IR H2 emission is discussed with detailed spectra given for a few planetary nebulae, and airborne observations of far-IR atomic lines are discussed. These two methods can discern photodissociation regions with warm gas and UV flux is fairly prominent. The use of resonance-absorption-line spectroscopy is also reviewed with respect to the analysis of the Na D lines, and thereby allows the measurement of integrated columns of material through the shell. The methods provide evidence for the notion that planetary nebulae consist of more than just ionized material; large amounts of neutral and molecular material are being confirmed, which has important implications for the mass-loss episode of the nebulae.
A Analysis of Neutral pi Mesons Produced in Minimum Bias 515 Gev/c Negative Pion Nuclear Collisions
NASA Astrophysics Data System (ADS)
Kuehler, John Fredrick
The results of an analysis of the minimum bias data sample collected from experiment E706 at Fermi National Accelerator Laboratory are reported on. The measured invariant differential cross sections, E{dsigma over d^3p}, for pi ^0 production from the interaction of a 515 GeV/c pi^- beam incident on the nuclear targets of Beryllium, pi^ - + {bf Be} to pi^0 + {bf X}, and Copper, pi ^- + {bf Cu} to pi^0 + {bf X}, are measured in terms of p_{t} (transverse momentum) over the range of.6 GeV/c <= p_{t } <= 2. GeV/c. Further, the cross sections for particle production off of nuclear targets can be parameterized as E{dsigmaover d^3p} = sigma_0A^{ alpha(p_{t})}, and the alpha(p_{t}) dependence of the differential cross section is reported on.
Evaluation of smallpox vaccines using variola neutralization.
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
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.
Children's Performance on and Attraction to an Activity as a Function of Masculine, Feminine or Neutral Labels on Sex-Role Preference.
ERIC Educational Resources Information Center
Montemayor, Raymond
Children six to eight years old each played a game which was labeled either sex-appropriate, sex-neutral, or sex-inappropriate. Measures of performance and attractiveness of the game were obtained. For both boys and girls, performance was highest when the game was labeled sex-appropriate, intermediate when no sex label was given, and lowest when…
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...
Spatial and temporal distribution of the neutral polymorphisms in the last ZFX intron: analysis of the haplotype structure and genealogy.
Jaruzelska, J; Zietkiewicz, E; Batzer, M; Cole, D E; Moisan, J P; Scozzari, R; Tavaré, S; Labuda, D
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.
Structural Analysis of Human and Macaque mAbs 2909 and 2.5B: Implications for the Configuration of the Quaternary Neutralizing Epitope of HIV-1 gp120
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
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.
A class of neutral functional differential equations.
NASA Technical Reports Server (NTRS)
Melvin, W. R.
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.
Characterization of a Novel Neutralizing Monoclonal Antibody Against Ebola Virus GP.
Reynard, Olivier; Volchkov, Viktor E
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.
Current Analysis Status for the Inclusive Neutral Current π0 Production Cross section Measurement with the NOvA Near Detector
NASA Astrophysics Data System (ADS)
Kalra, Daisy; Brunetti, Giulia
NOvA (NuMI Off-axis νe Appearance) experiment is designed to study long-baseline neutrino oscillations. It uses two detectors, the Near Detector (ND) at Fermilab and the Far Detector (FD) at a distance 810 km in Northern Minnesota. NOvA looks for the νe appearance at the FD using a narrow band νμ beam peaked at 2 GeV in energy. Neutral Current (NC) interactions with a π0 in the final state represents the main background in the νe appearance measurement. The π0 decay into two photons can fake the νe appearance either due to merging of two photon showers or one of the two photons escaping the detection. Therefore, a complete understanding of νμ induced NC interactions with a π0 in the final state is very important. It will also help in reducing the background uncertainties for current and future long-baseline neutrino oscillation experiments. We will present the status of the analysis related to the inclusive NC π0 production cross section measurement with the NOvA ND.
Content Analysis in Systems Engineering Acquisition Activities
shape requirements definitions for system upgrade or modification contracts and new baseline contracts. Finally, content analysis training and skill...back to the system designers, this information can then be used to shape requirements definition for system upgrade or modification contracts and new...Activity System Requirements Definition Ensuring the system requirements adequately reflect the stakeholder requirements Negotiating modifications to
Accession Medical Standards Analysis and Research Activity
Chief, Accession Medical Standards Analysis & Research Activity Li Yuanzhang, PhD Senior Statistician Department of Epidemiology David N...ORGANIZATION NAME(S) AND ADDRESS(ES) AMSARA, Department of Epidemiology , Division of Preventive Medicine Walter Reed Army Institute of Research 503... Epidemiology of Injury form the Assessment of Recruit Strength and Motivation study ARMS) and Program
Tautomerism in neutral histidine.
Bermúdez, Celina; Mata, Santiago; Cabezas, Carlos; Alonso, José L
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.
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.
Chen, Yixin; Li, Chuan; He, Delei; Cheng, Tong; Ge, Shengxiang; Shih, James Wai-Kuo; Zhao, Qinjian; Chen, Pei-Jer; Zhang, Jun; Xia, Ningshao
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.
Hippocampus and nucleus accumbens activity during neutral word recognition related to trait physical anhedonia in patients with schizophrenia: an fMRI study.
Lee, Jung Suk; Chun, Ji Won; Kang, Jee In; Kang, Dong-Il; Park, Hae-Jeong; Kim, Jae-Jin
Emotional memory dysfunction may be associated with anhedonia in schizophrenia. This study aimed to investigate the neurobiological basis of emotional memory and its relationship with anhedonia in schizophrenia specifically in emotional memory relate brain regions of interest (ROIs) including the amygdala, hippocampus, nucleus accumbens, and ventromedial prefrontal cortex. Fourteen patients with schizophrenia and 16 healthy subjects performed a word-image associative encoding task, during which a neutral word was presented with a positive, neutral, or control image. Subjects underwent functional magnetic resonance imaging while performing the recognition task. Correlation analyses were performed between the percent signal change (PSC) in the ROIs and the anhedonia scores. We found no group differences in recognition accuracy and reaction time. The PSC of the hippocampus in the positive and neutral conditions, and the PSC in the nucleus accumbens in the control condition, appeared to be negatively correlated with the Physical Anhedonia Scale (PAS) scores in patients with schizophrenia, while significant correlations with the PAS scores were not observed in healthy subjects. This study provides further evidences of the role of the hippocampus and nucleus accumbens in trait physical anhedonia and possible associations between emotional memory deficit and trait physical anhedonia in patients with schizophrenia.
Transition state of a SH3 domain detected with principle component analysis and a charge-neutralized all-atom protein model.
Mitomo, Daisuke; Nakamura, Hironori K; Ikeda, Kazuyoshi; Yamagishi, Akihiko; Higo, Junichi
The src SH3 domain has been known to be a two-state folder near room temperature. However, in a previous study with an all-atom model simulation near room temperature, the transition state of this protein was not successfully detected on a free-energy profile using two axes: the radius of gyration (R(g)) and native contact reproduction ratio (Q value). In this study, we focused on an atom packing effect to characterize the transition state and tried another analysis to detect it. To explore the atom packing effect more efficiently, we introduced a charge-neutralized all-atom model, where all of the atoms in the protein and water molecules were treated explicitly, but their partial atomic charges were set to zero. Ten molecular dynamics simulations were performed starting from the native structure at 300 K, where the simulation length of each run was 90 ns, and the protein unfolded in all runs. The integrated trajectories (10 x 90 = 900 ns) were analyzed by a principal component analysis (PCA) and showed a clear free-energy barrier between folded- and unfolded-state conformational clusters in a conformational space generated by PCA. There were segments that largely deformed when the conformation passed through the free-energy barrier. These segments correlated well with the structural core regions characterized by large phi-values, and the atom-packing changes correlated with the conformational deformations. Interestingly, using the same simulation data, no significant barrier was found in a free-energy profile using the R(g) and Q values for the coordinate axes. These results suggest that the atom packing effect may be one of the most important determinants of the transition state.
NASA Astrophysics Data System (ADS)
Goede, Adelbert; van de Sanden, Richard
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.
Bleach Neutralizes Mold Allergens
ERIC Educational Resources Information Center
Science Teacher, 2005
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…
Neutrality tests for sequences with missing data.
Ferretti, Luca; Raineri, Emanuele; Ramos-Onsins, Sebastian
Missing data are common in DNA sequences obtained through high-throughput sequencing. Furthermore, samples of low quality or problems in the experimental protocol often cause a loss of data even with traditional sequencing technologies. Here we propose modified estimators of variability and neutrality tests that can be naturally applied to sequences with missing data, without the need to remove bases or individuals from the analysis. Modified statistics include the Watterson estimator θW, Tajima's D, Fay and Wu's H, and HKA. We develop a general framework to take missing data into account in frequency spectrum-based neutrality tests and we derive the exact expression for the variance of these statistics under the neutral model. The neutrality tests proposed here can also be used as summary statistics to describe the information contained in other classes of data like DNA microarrays.
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…
Recombinant Sheep Pox Virus Proteins Elicit Neutralizing Antibodies
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.
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
Characteristics of HPV-Specific Antibody Responses Induced by Infection and Vaccination: Cross-Reactivity, Neutralizing Activity, Avidity and IgG Subclasses
Scherpenisse, Mirte; Schepp, Rutger M.; Mollers, Madelief; Meijer, Chris J. L. M.; Berbers, Guy A. M.; van der Klis, Fiona R. M.
Objectives In order to assess HPV-specific IgG characteristics, we evaluated multiple aspects of the humoral antibody response that will provide insight in the HPV humoral immune response induced by HPV infection and vaccination. Methods Cross-reactivity of HPV-specific antibodies induced by infection or vaccination was assessed with VLP16 or 18 inhibition using a VLP-based multiplex immunoassay (MIA) for HPV16, 18, 31, 33, 45, 52 and 58. HPV16/18 specific IgG1-4 subclasses and avidity were determined with the VLP-MIA in sera after HPV infection and after vaccination. Neutralizing antibodies were determined in a small subset of single-seropositive and multi-seropositive naturally derived antibodies. Results Naturally derived antibodies from single-positive sera were highly genotype-specific as homologue VLP-inhibition percentages varied between 78-94%. In multi-positive sera, cross-reactive antibodies were observed both within and between α7 and α9 species. After vaccination, cross-reactive antibodies were mainly species-specific. Avidity of vaccine-derived HPV-specific antibodies was 3 times higher than that of antibodies induced by HPV infection (p<0.0001). IgG1 and IgG3 were found to be the predominant subclasses observed after HPV infection and vaccination. In the small subset tested, the number of single-positive sera with neutralizing capacity was higher than of multi-positive sera. Conclusion Naturally derived HPV-specific antibodies from single-positive samples showed different characteristics in terms of cross-reactivity and neutralizing capacity compared with antibodies from multi-positive sera. Post-vaccination, HPV antibody avidity was approximately 3 times higher than antibody avidity induced by HPV infection. Therefore, antibody avidity might be a potential surrogate of protection. PMID:24058629
Perturbative studies of toroidal momentum transport using neutral beam injection modulation in the Joint European Torus: Experimental results, analysis methodology, and first principles modeling
Mantica, P.; Ferreira, J. S.; Salmi, A.; Strintzi, D.; Weiland, J.; Brix, M.; Giroud, C.; Corrigan, G.; Zastrow, K.-D.; Tardini, G.
Perturbative experiments have been carried out in the Joint European Torus [Fusion Sci. Technol. 53(4) (2008)] in order to identify the diffusive and convective components of toroidal momentum transport. The torque source was modulated either by modulating tangential neutral beam power or by modulating in antiphase tangential and normal beams to produce a torque perturbation in the absence of a power perturbation. The resulting periodic perturbation in the toroidal rotation velocity was modeled using time-dependent transport simulations in order to extract empirical profiles of momentum diffusivity and pinch. Details of the experimental technique, data analysis, and modeling are provided. The momentum diffusivity in the core region (0.2<{rho}<0.8) was found to be close to the ion heat diffusivity ({chi}{sub {phi}/{chi}i{approx}}0.7-1.7) and a significant inward momentum convection term, up to 20 m/s, was found, leading to an effective momentum diffusivity significantly lower than the ion heat diffusivity ({chi}{sub {phi}}{sup eff}/{chi}{sub i}{sup eff{approx}}0.4). These results have significant implications on the prediction of toroidal rotation velocities in future tokamaks and are qualitatively consistent with recent developments in momentum transport theory. Detailed quantitative comparisons with the theoretical predictions of the linear gyrokinetic code GKW [A. G. Peeters et al., Comput. Phys. Commun. 180, 2650 (2009)] and of the quasilinear fluid Weiland model [J. Weiland, Collective Modes in Inhomogeneous Plasmas (IOP, Bristol, 2000)] are presented for two analyzed discharges.
Antitumor resistance induced by zinostatin stimalamer (ZSS), a polymer-conjugated neocarzinostatin (NCS) derivative. I. Meth A tumor eradication and tumor-neutralizing activity in mice pretreated with ZSS or NCS.
Masuda, E; Maeda, H
Zinostatin stimalamer (ZSS) is a new anticancer agent derived from neocarzinostatin (NCS), which is synthesized by conjugation of one molecule of NCS and two molecules of poly(styrene-co-maleic acid). ZSS exhibited potent in vitro and in vivo antitumor activity in preclinical experiments, and a clinical trial of the intra-arterial administration of ZSS with iodized oil on hepatocellular carcinoma showed potent antitumor activity. We investigated the effect of ZSS and NCS on antitumor resistance and found that pretreatment with either drug suppressed the growth of MethA tumors in Balb/c mice and induced tumor eradication when given separately by single administration at therapeutic doses between 1 day and 4 weeks before tumor transplantation. The findings that the cytocidal activity of these drugs was not detected in vivo at the time of tumor transplantation and that tumor regression was preceded by a period of transient growth suggested that tumor regression was due to host-mediated antitumor activity induced by these drugs. Pretreatment with ZSS or NCS also suppressed the growth of Colon 26 carcinoma and Sarcoma 180. The finding that NCS showed the same effect as ZSS suggests that poly(styrene-comaleic acid) is not essential for the induction of host-mediated antitumor activity. Furthermore, apo-ZSS, which lacks cytocidal activity, did not induce antitumor activity. From this, it is suggested that the cytocidal effect of ZSS involves the induction of host-mediated antitumor resistance. In athymic Balb/c nu/nu mice, pretreatment with ZSS or NCS did not induce tumor eradication, suggesting that mature T lymphocytes play an important role in tumor eradication. Challenging MethA was rejected without transient growth in mice that had been cured of MethA, but challenging Colon 26 was not, showing that anti-MethA resistance was augmented selectively in the MethA-eradicated mice. Splenocytes from MethA-bearing mice pretreated with the drug showed tumor-neutralizing
Zeman, Frank S; Keith, David W
Reducing greenhouse gas emissions from the transportation sector may be the most difficult aspect of climate change mitigation. We suggest that carbon neutral hydrocarbons (CNHCs) offer an alternative pathway for deep emission cuts that complement the use of decarbonized energy carriers. Such fuels are synthesized from atmospheric carbon dioxide (CO2) and carbon neutral hydrogen. The result is a liquid fuel compatible with the existing transportation infrastructure and therefore capable of a gradual deployment with minimum supply disruption. Capturing the atmospheric CO2 can be accomplished using biomass or industrial methods referred to as air capture. The viability of biomass fuels is strongly dependent on the environmental impacts of biomass production. Strong constraints on land use may favour the use of air capture. We conclude that CNHCs may be a viable alternative to hydrogen or conventional biofuels and warrant a comparable level of research effort and support.
Pack, Michael Vern
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.
Antihypertensive neutral lipid
Snyder, Fred L.; Blank, Merle L.
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.
Exercise Equipment: Neutral Buoyancy
NASA Technical Reports Server (NTRS)
Shackelford, Linda; Valle, Paul
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.
Antihypertensive neutral lipid
Snyder, F.L.; Blank, M.L.
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.
Neutral particle beam intensity controller
Dagenhart, W.K.
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.
Thermal neutral format based on the step technology
NASA Technical Reports Server (NTRS)
Almazan, P. Planas; Legal, J. L.
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.
Comparative Sensitivity Analysis of Muscle Activation Dynamics
Rockenfeller, Robert; Günther, Michael; Schmitt, Syn; Götz, Thomas
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
Neutron activation analysis in archaeological chemistry
Harbottle, G.
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.
Neutrality between Government and Religion.
ERIC Educational Resources Information Center
Mawdsley, Ralph D.
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…
Cross-Reactivity of Neutralizing Antibodies among Malignant Catarrhal Fever Viruses
Taus, Naomi S.; Cunha, Cristina W.; Marquard, Jana; O'Toole, Donal; Li, Hong
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
Analysis of DOE international environmental management activities
Ragaini, R.C.
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.
Peptides in Seminal Fluid and Their Role in Infertility: A Potential Role for Opiorphin Inhibition of Neutral Endopeptidase Activity as a Clinically Relevant Modulator of Sperm Motility
Davies, Kelvin P.; Neal-Perry, Genevieve S.
Infertility is a devastating medical condition that adversely affects emotional health and well-being of couples who desire pregnancy and parenthood. The overall demographic data suggest that the indication for more than one-third of assisted reproductive technology cycles performed in the United States includes male factor infertility. There is increasing recognition of the role that peptides present in seminal plasma have in determining sperm motility. Several recent studies suggest that peptidases, such as neutral endopeptidase (NEP) and aminopeptidase N (APN), impose significant adverse effects on sperm motility. Interestingly, several recent studies demonstrate that there is an endogenous NEP/APN inhibitor peptide called opiorphin in human seminal plasma. Our pilot studies suggest opiorphin promotes sperm motility and may positively influence sperm motility parameters in some cases of males infertility characterized by asthenozoospermia. PMID:24855109
Characterization of human single-chain antibodies against highly pathogenic avian influenza H5N1 viruses: mimotope and neutralizing activity.
Yang, Jiupian; Yoshida, Reiko; Kariya, Yuki; Zhang, Xu; Hashiguchi, Shuhei; Nakashima, Toshihiro; Suda, Yasuo; Takada, Ayato; Ito, Yuji; Sugimura, Kazuhisa
The development of new therapeutic targets and strategies to control highly pathogenic avian influenza (HPAI) H5N1 virus infection in humans is urgently needed. Neutralizing recombinant human antibodies would provide important agents for immunotherapy on human H5N1 virus infection and definition of the critical mimotope for vaccine development. In this study, we have characterized an anti-H5-specific scFv clone, 3D1 from the human-scFv-displaying phage library. 3D1 blocked the binding of H5-Fc to MDCK cells in flow cytometry and neutralized H5N1 subtype influenza A viruses in a microneutralization assay. Employing a peptide-displaying phage library, Ph.D-12, the mimotope was determined to be at #128-131 and #204-211 of H5, which are silic acid-binding regions. In consistency with this result, 3D1 binds the recombinant sugar-binding domain (#50G-#272E) produced by a baculovirus vector. The 3D1 antibody employs the germline gene VH1-23. As this antibody is the first human anti-H5 scFv clearly defined on the sugar-binding epitope, it allows us to investigate the influence of amino acid substitutions in this region on the determination of the binding specificity to either sialic acid α2,6-galactose (SA α2,6Gal) or sialic acid α2,3-galactose (SA α2,3Gal) providing new insight for the development of effective H5N1 pandemic vaccines.
Exchanging physically effective neutral detergent fiber does not affect chewing activity and performance of late-lactation dairy cows fed corn and sugarcane silages.
Sá Neto, A; Bispo, A W; Junges, D; Bercht, A K; Zopollatto, M; Daniel, J L P; Nussio, L G
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.
NASA Astrophysics Data System (ADS)
Lyon, M.; Rolston, S. L.
By photoionizing samples of laser-cooled atoms with laser light tuned just above the ionization limit, plasmas can be created with electron and ion temperatures below 10 K. These ultracold neutral plasmas have extended the temperature bounds of plasma physics by two orders of magnitude. Table-top experiments, using many of the tools from atomic physics, allow for the study of plasma phenomena in this new regime with independent control over the density and temperature of the plasma through the excitation process. Characteristic of these systems is an inhomogeneous density profile, inherited from the density distribution of the laser-cooled neutral atom sample. Most work has dealt with unconfined plasmas in vacuum, which expand outward at velocities of order 100 m/s, governed by electron pressure, and with lifetimes of order 100 μs, limited by stray electric fields. Using detection of charged particles and optical detection techniques, a wide variety of properties and phenomena have been observed, including expansion dynamics, collective excitations in both the electrons and ions, and collisional properties. Through three-body recombination collisions, the plasmas rapidly form Rydberg atoms, and clouds of cold Rydberg atoms have been observed to spontaneously avalanche ionize to form plasmas. Of particular interest is the possibility of the formation of strongly coupled plasmas, where Coulomb forces dominate thermal motion and correlations become important. The strongest impediment to strong coupling is disorder-induced heating, a process in which Coulomb energy from an initially disordered sample is converted into thermal energy. This restricts electrons to a weakly coupled regime and leaves the ions barely within the strongly coupled regime. This review will give an overview of the field of ultracold neutral plasmas, from its inception in 1999 to current work, including efforts to increase strong coupling and effects on plasma properties due to strong coupling.
Pulsed field sample neutralization
Appelhans, Anthony D.; Dahl, David A.; Delmore, James E.
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.
Duesing, G.; Altmann, H.; Falter, H.; Goede, A.; Haange, R.; Hemsworth, R.S.; Kupschus, P.; Stork, D.; Thompson, E.
The development of the neutral injection (NI) system for the Joint European Torus and its status in 1985 are reported. First the system parameters are discussed and the layout is described, followed by a summary of the physics design calculations, the development, production, and testing of the components and the subsystem assembly. The system commissioning is presented, including a description of the function and the realization of the NI test bed. A summary of performance predictions for 80-keV beam heating experiments, and of the experimental evidence on balanced versus coinjection, is presented. The operational experience with the first injector and the plasma physics results obtained so far are summarized.
Coherent and neutral pion production results from MINERνA
Palomino, J. L.; Higuera, A.
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.
Profiling the neutralizing antibody response in chronically HIV-1 CRF07_BC-infected intravenous drug users naïve to antiretroviral therapy
Hu, Xintao; Hu, Yuanyuan; Zhao, Chunhong; Gao, Hongmei; Greene, Kelli M.; Ren, Li; Ma, Liying; Ruan, Yuhua; Sarzotti-Kelsoe, Marcella; Montefiori, David C.; Hong, Kunxue; Shao, Yiming
Characterizing neutralizing antibody (NAb) responses in individuals infected with diverse HIV-1 strains is necessary to reveal the novel targets for regional preventive and therapeutic strategies development. We evaluated the prevalence, breadth, and potency of NAb responses in 98 CRF07_BC-infected individuals using a large, multi-subtype panel of 30 tier 2-3 Env-pseudotyped viruses. Furthermore, we compared the neutralization pattern of CRF07_BC-infected people with that of subtype B'-infected individuals in China. Of the 98 plasma samples tested, 18% neutralized more than 80% of viruses in the panel, and 53% neutralized more than 50%, suggesting the presence of broadly NAbs in these individuals. A preferential intra-subtype neutralization of CRF07_BC was found. Notably, CRF07_BC-infected individuals generated higher neutralization titers against intra-subtype viruses than subtype B'-infected individuals with longer infection length. However, subtype B'-infected individuals mounted broader neutralization responses against inter-subtype viruses than CRF07_BC infection with shorter infection time, indicating the transition from narrow autologous to broad heterologous neutralization over time. Neutralization activity of the top six plasmas from each cohort was attributable to IgG fraction, and half of them developed CD4 binding site antibody reactivity. Heatmap analysis identified three statistically robust clusters of plasmas that offer valuable resources for further in-depth virological and immunological study. PMID:28387330
Characterization of Neutralizing Antibodies and Identification of Neutralizing Epitope Mimics on the Clostridium botulinum Neurotoxin Type A
Wu, Han-Chung; Yeh, Chia-Tsui; Huang, Yue-Ling; Tarn, Lih-Jeng; Lung, Chien-Cheng
Clostridium botulinum neurotoxin type A (BTx-A) is known to inhibit the release of acetylcholine at the neuromuscular junctions and synapses and to cause neuroparalysis and death. In this study, we have identified two monoclonal antibodies, BT57-1 and BT150-3, which protect ICR mice against lethal doses of BTx-A challenge. The neutralizing activities for BT57-1 and BT150-3 were 103 and 104 times the 50% lethal dose, respectively. Using immunoblotting analysis, BT57-1 was recognized as a light chain and BT150-3 was recognized as a heavy chain of BTx-A. Also, applying the phage display method, we investigated the antibodies' neutralizing B-cell epitopes. These immunopositive phage clones displayed consensus motifs, Asp-Pro-Leu for BT57-1 and Cys-X-Asp-Cys for BT150. The synthetic peptide P4M (KGTFDPLQEPRT) corresponded to the phage-displayed peptide selected by BT57-1 and was able to bind the antibodies specifically. This peptide was also shown by competitive inhibition assay to be able to inhibit phage clone binding to BT57-1. Aspartic acid (D5) in P4M was crucial to the binding of P4M to BT57-1, since its binding activity dramatically decreased when it was changed to lysine (K5). Finally, immunizing mice with the selected phage clones elicited a specific humoral response against BTx-A. These results suggest that phage-displayed random-peptide libraries are useful in identifying the neutralizing epitopes of monoclonal antibodies. In the future, the identification of the neutralizing epitopes of BTx-A may provide important information for the identification of the BTx-A receptor and the design of a BTx-A vaccine. PMID:11425742
Self-assembly of heterobimetallic neutral macrocycles incorporating ferrocene spacer groups: spectroelectrochemical analysis of the double two-electron oxidation of a molecular rectangle.
Das, Neeladri; Arif, Atta M; Stang, Peter J; Sieger, Monika; Sarkar, Biprajit; Kaim, Wolfgang; Fiedler, Jan
Two Pt(4)-Fe(2) mixed-metal neutral assemblies, 4 and 5, incorporating four bis(triethylphosphine)platinum(II) centers, two flexible bridging 1,1'-ferrocenedicarboxylates, and two rigid 2,9-phenanthrenediyl (4) or 1,8-anthracenediyl (5) bridges, have been synthesized. X-ray characterization of 4 and 5 reveals the formation of discrete and highly symmetrical heterobimetallic neutral species possessing a rhomboidal and rectangular shape, respectively. The rectangular molecules, 5, could be reversibly oxidized in two two-electron steps, separated by 0.21 V. Spectroelectrochemistry in the UV-vis-NIR region confirms the ferrocene groups as primary oxidation sites; however, the intermediate 5(2+) is EPR silent even at 4 K due to enhanced EPR relaxation involving the oxidizable 1,8-anthracenediyl linkers.
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.
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).
Ergonomically neutral arm support system
Siminovitch, Michael J; Chung, Jeffrey Y; Dellinges, Steven; Lafever, Robin E
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.
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.
Chebabhi, Ali; Fellah, Mohammed Karim; Kessal, Abdelhalim; Benkhoris, Mohamed F
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.
Retinol binding protein and vitamin D associations with serum antibody isotypes, serum influenza virus-specific neutralizing activities and airway cytokine profiles.
Jones, B G; Oshansky, C M; Bajracharya, R; Tang, L; Sun, Y; Wong, S S; Webby, R; Thomas, P G; Hurwitz, J L
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.
Hydrolysis of ionized deoxycholic acid in the aqueous phase and rate analysis for transfer of neutralized deoxycholic acid into the benzene phase across the benzene/water interface.
Ohno, Ryo; Nakamura, Shohei; Moroi, Yoshikiyo; Isoda-Yamashita, Teruyo
Sodium deoxycholate in water dissociates into sodium cation and deoxycholate anion in the aqueous phase, and then, the latter anions partially hydrolyze to form deionized deoxycholic acids. The acids move into the benzene phase, when liquid benzene is placed upon the aqueous phase, and finally the partition equilibrium is reached. The above processes were traced by pH change in the aqueous phase by a pH meter or the change in [OH-] with time, from which the rate for transfer of neutralized acid to the organic phase was analyzed. From the trace, the rate constants for hydrolysis of acid anion ( kf), neutralization of acid ( kb), transfer of neutralized acid from the aqueous phase to the organic phase ( kin*), and its back-transfer from the organic phase to the aqueous phase ( kut*) were evaluated; kf = 2.18 x 10 (-4) mol (-1) dm (3) min (-1), kb = 1.24 x 10 (5) mol (-1) dm (3) min (-1), kin* = 4.06 x 10 (-1) min (-1) cm (-2), and kout*) = 8.00 x 10 (-2) min (-1) cm (-2). The above values are supported by the partition constant of deoxycholic acid between the benzene phase and the aqueous phase.
Viewing perspective in energetic neutral atom intensity
NASA Astrophysics Data System (ADS)
Zheng, Yihua; Lui, Anthony T. Y.; Fok, Mei-Ching
Through interspacecraft comparison of energetic neutral oxygen (ENO) intensity from two different vantage points provided by IMAGE and Geotail, Lui et al. (2005) showed that viewing perspective plays a very important role in the observed ENO intensity level during a magnetic storm period. Motivated by the findings of Lui et al. (2005), we investigate how viewing perspective influences energetic neutral atom emissions from a modeling perspective. The main results of this paper are that (1) our simulation results, based upon O+ ion fluxes from the Comprehensive Ring Current Model and the subsequent ENO calculation, reproduce the total differential ENO intensity obtained from two spacecraft to a reasonable degree and (2) further analysis of our results indicates that pitch angle anisotropy in ring current ion flux, a crucial physical quantity in ring current dynamics, is one major contributor to the difference in energetic neutral atom intensity from different viewing perspectives.
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.
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.
A multistate analysis of active life expectancy.
Rogers, A; Rogers, R G; Branch, L G
With today's lower mortality rates, longer expectations of life, and new medical technologies, the nation's health policy focus has shifted from emphasis on individual survival to emphasis on personal health and independent living. Using longitudinal data sets and new methodological techniques, researchers have begun to assess active life expectancies, estimating not only how long a subpopulation can expect to live beyond each age, but what fractions of the expected remaining lifetime will be lived as independent, dependent, or institutionalized. New ideas are addressed, applying recently developed multistate life table methods to Waves One and Two of the Massachusetts Health Care Panel Study. Expectations of active life are presented for those 65 and older who initially are in one of two functional states of well-being. Included are expectations of life, for those, for example, who were independent and remained so, or those who were dependent and became independent. Although public health officials are concerned about the number of elderly who cease being independent, preliminary analysis shows that a significant number of the dependent elderly regain their independence, a situation which needs to be addressed in health care planning.
Neutron activation analysis of some building materials
NASA Astrophysics Data System (ADS)
Salagean, M. N.; Pantelica, A. I.; Georgescu, I. I.; Muntean, M. I.
Concentrations of As, Au, Ba, Br, Ca, Ce, Co, Cr, Cs, Eu, Fe, Hf, K, La, Lu, Mo, Na, Nd, Rb, Sb, Sc, Sr, Ta, Tb, Th, U. Yb, W and Zn in seven Romanian building materials were determined by the Instrumental Neutron Activation Analysis (INAA) method using the VVR-S Reactor of NIPNE- Bucharest. Raw matarials used in cement obtaining ≈ 75% of limestone and ≈ 25% of clay, cement samples from three different factories, furnace slag, phosphogypsum, and a type of brick have been analyzed. The brick was compacted from furnace slay, fly coal ash, phosphogypsum, lime and cement. The U, Th and K concentrations determined in the brick are in agreement with the natural radioactivity measurements of226Ra,232Th and40K. These specific activities were found about twice and 1.5 higher than the accepted levels in the case of226Ra and232Th, as well as40K, respectively. By consequence, the investigated brick is considered a radioactive waste. The rather high content of Co, Cr, K, Th, and Zh in the brick is especially due to the slag and fly ash, the main componets. The presence of U, Th and K in slag is mainly correlated with the limestone and dolomite as fluxes in matallurgy.
Active polarimeter optical system laser hazard analysis.
Augustoni, Arnold L.
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.
Neutral polypropylene laser welding
NASA Astrophysics Data System (ADS)
Mandolfino, Chiara; Lertora, Enrico; Gambaro, Carla
The joining of polymeric materials is a technology used in many industrial applications, from transport to telecommunications and the medical sector. A new technology for the joining of polymers is the laser welding process. In particular, fibre laser welding is a flexible technology which allows high process speed and the realization of good quality joints. Despite its application becoming more widespread in the production of assemblies of high precision, the application of laser technology for the welding of polymers has not been the subject of many studies up to now. This study focused on the welding of neutral polypropylene. The window process parameter was identified, without the use of additives to increase radiation absorption, and a mechanical characterization was conducted in order to evaluate the quality of the joints realized.
Protection of mice against the highly pathogenic VVIHD-J by DNA and fowlpox recombinant vaccines, administered by electroporation and intranasal routes, correlates with serum neutralizing activity.
Bissa, Massimiliano; Quaglino, Elena; Zanotto, Carlo; Illiano, Elena; Rolih, Valeria; Pacchioni, Sole; Cavallo, Federica; De Giuli Morghen, Carlo; Radaelli, Antonia
The control of smallpox was achieved using live vaccinia virus (VV) vaccine, which successfully eradicated the disease worldwide. As the variola virus no longer exists as a natural infection agent, mass vaccination was discontinued after 1980. However, emergence of smallpox outbreaks caused by accidental or deliberate release of variola virus has stimulated new research for second-generation vaccine development based on attenuated VV strains. Considering the closely related animal poxviruses that also arise as zoonoses, and the increasing number of unvaccinated or immunocompromised people, a safer and more effective vaccine is still required. With this aim, new vectors based on avian poxviruses that cannot replicate in mammals should improve the safety of conventional vaccines, and protect from zoonotic orthopoxvirus diseases, such as cowpox and monkeypox. In this study, DNA and fowlpox (FP) recombinants that expressed the VV L1R, A27L, A33R, and B5R genes were generated (4DNAmix, 4FPmix, respectively) and tested in mice using novel administration routes. Mice were primed with 4DNAmix by electroporation, and boosted with 4FPmix applied intranasally. The lethal VVIHD-J strain was then administered by intranasal challenge. All of the mice receiving 4DNAmix followed by 4FPmix, and 20% of the mice immunized only with 4FPmix, were protected. The induction of specific humoral and cellular immune responses directly correlated with this protection. In particular, higher anti-A27 antibodies and IFNγ-producing T lymphocytes were measured in the blood and spleen of the protected mice, as compared to controls. VVIHD-J neutralizing antibodies in sera from the protected mice suggest that the prime/boost vaccination regimen with 4DNAmix plus 4FPmix may be an effective and safe mode to induce protection against smallpox and poxvirus zoonotic infections. The electroporation/intranasal administration routes contributed to effective immune responses and mouse survival.
Decreasing sensitivity to RANTES (regulated on activation, normally T cell-expressed and -secreted) neutralization of CC chemokine receptor 5-using, non-syncytium-inducing virus variants in the course of human immunodeficiency virus type 1 infection.
Koning, Fransje A; Kwa, David; Boeser-Nunnink, Brigitte; Dekker, Jos; Vingerhoed, Jose; Hiemstra, Harry; Schuitemaker, Hanneke
In approximately half of human immunodeficiency virus (HIV) type 1-infected individuals, the development of CXC chemokine receptor 4-using, syncytium-inducing (SI) virus variants precedes a rapid progression to acquired immunodeficiency syndrome (AIDS). In other individuals, only CC chemokine receptor 5-using (R5), non-SI (NSI) virus variants are present throughout infection. These individuals may be either long-term survivors (LTSs) or rapid progressors. The basis for this variable disease progression in individuals with only R5 virus variants is not yet fully understood. In this study, the beta-chemokine sensitivity of biological HIV-1 clones isolated from 13 individuals who harbored only R5, NSI virus variants (7 LTSs and 6 progressors) was investigated. We found a statistically significant decrease in sensitivity of virus variants to RANTES (regulated on activation, normally T cell-expressed and -secreted) neutralization during the course of progressive infection, but not during follow-up of LTSs. Our data suggest that a role exists for RANTES neutralization sensitivity of HIV-1 in AIDS pathogenesis.
Transient Photochemistry of Neutral Red.
ascorbic acid system to 50 successive flashes, indicating that no ground state neutral red is permanently converted to leuco dye . Since leuco neutral...complete regeneration of ground state neutral red in this pH range in the present study suggests that formation of leuco dye is not significant. The second...radical disproportionation step is followed by a slower step which converts leuco dye to semireduced radical. Because coupling the two steps
Transient ion neutralization by electrons.
NASA Technical Reports Server (NTRS)
Wilhelm, H. E.
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.
Correlation of serum triglyceride and its reduction by omega-3 fatty acids with lipid transfer activity and the neutral lipid compositions of high-density and low-density lipoproteins.
Pownall, H J; Brauchi, D; Kilinç, C; Osmundsen, K; Pao, Q; Payton-Ross, C; Gotto, A M; Ballantyne, C M
Serum triglyceride (TG) and high-density lipoprotein cholesterol (HDL-C) concentrations are inversely correlated and mechanistically linked by means of lipid transfer activities. Phospholipid transfer activity (PLTA) moves phospholipids among serum lipoproteins; cholesteryl ester transfer activity (CETA), which exchanges cholesteryl esters (CE) and TG among lipoproteins, is stimulated by nonesterified fatty acids (NEFA). The aims of this study were (a) to develop a quantitative model that correlates the neutral lipid (NL = CE + TG) compositions of HDL and LDL with serum TG concentration; (b) identify the serum lipid determinants of CETA and PLTA, and; (c) identify the effects of serum TG reductions on the neutral lipid compositions of HDL and LDL, serum NEFA concentrations, and on PLTA and CETA. These aims were addressed in 40 hypertriglyceridemic subjects before and after treatment with an 85% concentrate of omega-3 fatty acids (Omacor) and in 16 untreated normolipidemic subjects. In vivo, the NL compositions of LDL and HDL were described by a mathematical model having the form of adsorption isotherms: HDL - (TG/NL) = (0.90 +/- 0.07) serum TG/(7.0 +/- 1.2 mmol/l + serum TG) and LDL - (TG/NL) = (0.65 +/- 0.08) serum TG/(4.9 +/- 1.5 mmol/l + serum TG). Reduction of serum TG was associated with reductions in HDL - (TG/NL), serum NEFA concentration, and serum CETA but not PLTA. These data suggest that both hypertriglyceridemia and the attendant elevated serum CETA but not PLTA are determinants of HDL and LDL composition and structure and that serum TG concentrations are good predictors of the NL compositions of HDL and LDL.
Kunkel, W.B.
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
NASA Astrophysics Data System (ADS)
Goede, A. P. H.
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
A standard immunoglobulin preparation produced from bovine colostra shows antibody reactivity and neutralization activity against Shiga-like toxins and EHEC-hemolysin of Escherichia coli O157:H7.
Lissner, R; Schmidit, H; Karch, H
Enterohemorrhagic Escherichia coli (EHEC) causes a variety of clinical conditions, the most important being hemorrhagic colitis and hemolytic uremic syndrome. A curative therapy of EHEC diseases is not yet feasible. This study investigates the antibody reactivity of Lactobin, a standardized immunoglobulin (Ig) preparation, obtained from the colostra of non-immunized cows. Three different batches of Lactobin exhibited equally high titers of specific antibodies against Shiga-like toxins (SLTs, verocytotoxins) and EHEC hemolysin (EHEC-Hly) produced by E. coli O157. In addition, Lactobin blocked the cytotoxic effect of SLT-I and SLT-II on Vero cell monolayers and inhibited the cytolytic effects of EHEC-Hly on human erythrocytes. Since Lactobin contains high levels of antibodies and neutralizing activity against important virulence factors of EHEC O157, this drug has potential use in the treatment of diarrhea and the prevention of EHEC-associated hemolytic uremic syndrome.
[Distribution pattern of neutral sugar in forest soils along an altitude gradient in Changbai Mountains, Northeast China].
Tian, Qiu-Xiang; Zhang, Bin; He, Hong-Bo; Zhang, Xu-Dong; Cheng, Wei-Xin
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.
Eulogy for a neutron activation analysis facility
Lepel, E.A.
A relatively inexpensive facility for neutron activation analysis (NAA) was developed in the early 1970s at Pacific Northwest National Laboratory (PNNL). With the availability of large {sup 252}Cf sources, a subcritical facility was designed that could contain up to 100 mg of {sup 252}Cf (T{sub 1/2} = 2.645 yr and a spontaneous fission yield of 2.34 x 10{sup 9} n/s{center_dot}mg{sup {minus}1}). The {sup 252}Cf source was surrounded by a hexagonal array of {sup 235}U enriched fuel rods, which provided a 10- to 20-fold multiplication of the neutrons emitted from the {sup 252}Cf source. This assembly was located near the bottom of a 1.52-m-diam x 6.10-m-deep water-filled pool. The Neutron Multiplier Facility (NMF) was operational from November 1977 to April 1998--a period of 20.4 yr. The NMF began operation with {approximately}100 mg of {sup 252}Cf, and because of decay of the {sup 252}Cf, it had decreased to 0.34 mg at the time of shutdown. Decommissioning of the NMF began April 1998 and was completed in October 1999.
Signs of neutralization in a redundant gene involved in homologous recombination in Wolbachia endosymbionts.
Badawi, Myriam; Giraud, Isabelle; Vavre, Fabrice; Grève, Pierre; Cordaux, Richard
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.
Signs of Neutralization in a Redundant Gene Involved in Homologous Recombination in Wolbachia Endosymbionts
Badawi, Myriam; Giraud, Isabelle; Vavre, Fabrice; Grève, Pierre; Cordaux, Richard
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
Is science metaphysically neutral?
Fry, Iris
This paper challenges the claim that science is metaphysically neutral upheld by contenders of the separation of peacefully co-existent science and religion and by evolutionary theists. True, naturalistic metaphysical claims can neither be refuted nor proved and are thus distinct from empirical hypotheses. However, metaphysical assumptions not only regulate the theoretical and empirical study of nature, but are increasingly supported by the growing empirical body of science. This historically evolving interaction has contributed to the development of a naturalistic worldview that renounces the necessity of a transcendent god and of purposeful design. The thesis presented here differs not only from the claims of the "separatists" and of evolutionary theists. In pointing to the metaphysical aspects of science, I also criticize the failure of some evolutionary naturalists to distinguish between empirical and metaphysical contentions. Most important, based on the examination of science suggested here, creationists' false accusation that science is only a naturalistic dogma is refuted. Finally, the difficulties involved in the position endorsed here for the public support of evolution are acknowledged, taking into account the high religious profile of the American society and the social and political context in the US and in other countries.
Weak neutral current chemistry
Mohan, R.
Metal cluster organic complexes, neither atomic nor solid but in analogy to atomic nuclei and to mesoscopic systems, have unusual dynamics and catalytic properties. Organo-metal clusters as quintessence prebiotic enzymes could have originated the homochirality of the molecules from achiral precursors, controlled from the atomic-nucleus, with the initial product itself serving subsequently as chiral auxiliary transferring and amplifying the chirality in the autocatalytic process now. High resolution spectroscopic studies of diatomic molecules beginning now may lead to upper estimates of the interaction strength of weak neutral currents (WNG) with valence electrons of metal clusters and suggest kinetic pathways to dynamic symmetry breaking in the asymmetric synthesis of chiral molecules. An estimate of 10{sup {minus}5} kT (thousand times larger than for radiolysis) for the parity violating energy (PVE) could be sufficient to run an entropy driven spin-catalyzed asymmetric synthesis. Expect then, wherever there are metal clusters in interstellar dust or under the sea chiral molecular production. {copyright} {ital 1996 American Institute of Physics.}
Weak neutral current chemistry
NASA Astrophysics Data System (ADS)
Mohan, R.
Metal cluster organic complexes, neither atomic nor solid but in analogy to atomic nuclei and to mesoscopic systems, have unusual dynamics and catalytic properties. Organo-metal clusters as quintessence prebiotic enzymes could have originated the homochirality of the molecules from achiral precursors, controlled from the atomic-nucleus, with the initial product itself serving subsequently as chiral auxiliary transferring and amplifying the chirality in the autocatalytic process now. High resolution spectroscopic studies of diatomic molecules beginning now may lead to upper estimates of the interaction strength of weak neutral currents (WNG) with valence electrons of metal clusters and suggest kinetic pathways to dynamic symmetry breaking in the asymmetric synthesis of chiral molecules. An estimate of 10-5 kT (thousand times larger than for radiolysis) for the parity violating energy (PVE) could be sufficient to run an entropy driven spin-catalyzed asymmetric synthesis. Expect then, wherever there are metal clusters in interstellar dust or under the sea chiral molecular production.
[Neutralizing Monoclonal and Chimeric Antibodies to Human IFN-γ].
Larina, M V; Aliev, T K; Solopova, O N; Pozdnyakova, L P; Korobova, S V; Yakimov, S A; Sveshnikov, P G; Dolgikh, D A; Kirpichnikov, M P
Autoiminune disorders are chronic diseases characterized by abnormal immune response directed against self-antigens that leads to tissue damage and violation of its normal functioning. Such diseases often result in disability or even death of patients. Nowadays a number of monoclonal antibodies to pro-inflammatory cytokines and their receptors are successfully used for the targeted treatment of autoimmune diseases. One of the perspective targets in autoimmune disease therapy is interferon gamma, a key cytokine in Th1 cells differentiation, activation of macrophages, and inflammation. In the present work, 5 monoclonal antibodies to human IFN-γ were obtained. For the development of potential therapeutic agent, we have performed neutralizing activity and affinity analysis of the antibodies. Based on the data obtained, the monoclonal antibody F1 was selected. This antibody has a dissociation constant 1.7 x 10(-9) M and IC90 = 8.9 ± 2.0 nM measured upon antibody inhibition of the IFN-γ-induced HLA-DR expression on the surface of U937 cells. We have constructed a bicistronic vector for the production of recombinant chimeric Fab fragment F1 chim in E. coli cells. The recombinant chimeric Fab fragment Fl chim neutralizes IFN-γ activity in vitro and has a dissociation constant 1.8 x 10(-9) M.
Fine mapping of sequential neutralization epitopes on the subunit protein VP8 of human rotavirus.
Kovacs-Nolan, Jennifer; Yoo, Dongwan; Mine, Yoshinori
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
H7N9 influenza virus neutralizing antibodies that possess few somatic mutations
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.
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
Neutralization tests on the SERT 2 spacecraft
NASA Technical Reports Server (NTRS)
Kerslake, W. R.; Domitz, S.
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.
The Effects of Plasma-Neutral Interactions on Neutral Dynamics
NASA Astrophysics Data System (ADS)
Hsu, V.; Thayer, J. P.
Plasma-neutral interactions are fundamental to the structure and behavior of the neutral thermosphere. This interaction, primarily through ion-neutral collisions, ties electrodynamics with hydrodynamics requiring a fully coupled ionosphere - thermosphere model to simulate and dissect the sequence of responses that occur in the neutral gas when a change occurs in the ionosphere. In particular, changes in the ion drag force prompt a hydrodynamic response that will alter several properties of the thermosphere, including neutral winds. Here, the fully coupled National Center for Atmospheric Research Thermosphere-Ionosphere Electrodynamics General Circulation Model (NCAR TIEGCM) is used to evaluate how changes in mechanical coupling, through the ion drag force, alter thermosphere properties, with a focus on thermospheric neutral winds. The equatorial thermosphere anomaly (ETA) produces a transient wind system, and a dissection of the hydrodynamic processes responsible for its formation will be used to demonstrate the causal structure in neutral gas response to a change in field-aligned ion drag force. This well-behaved response elucidates processes that must be occurring in other regions of the thermosphere where more significant changes in the ion drag force occur.
Intraprostatic injection of neutralized zinc in rats
Fahim, M.S.; Wang, M.; Sutcu, M.F.; Fahim, Z.; Safron, J.A.; Ganjam, V.K. Xian Medical University )
Zinc has been implicated in steroid endocrinology of the prostate gland. The conversion of testosterone to dihydrotestosterone (DHT) by 5{alpha}-reductase enzyme is believed to express androgenic responses in the prostate. To note the effect of neutralized zinc on the prostate, 50 sexually mature rats, weighing 325 {plus minus} 20 grams, were divided into 5 groups as follows: (1) control, (2) sham, (3) castrated, (4) injected intraprostatically with 10 mg. neutralized zinc, and (5) injected intraprostatically with 20 mg. neutralized zinc. Results in the treated groups indicated significant reduction of prostate weights, 12% and 53% and histologically normal prostate; no significant change in weight and histological structure of testes, epididymides, and seminal vesicles; significant reduction in 5{alpha}-reductase activity and total protein and DNA concentrations in prostate tissue; and no significant effect on progeny of treated animals. These results suggest that direct application of neutralized zinc to the prostate offers a new modality for treatment of prostatitis without affecting spermatogenesis and testosterone production.
Pairwise transcriptomic analysis of the interactions between the ectomycorrhizal fungus Laccaria bicolor S238N and three beneficial, neutral and antagonistic soil bacteria.
Deveau, Aurélie; Barret, Matthieu; Diedhiou, Abdala G; Leveau, Johan; de Boer, Wietse; Martin, Francis; Sarniguet, Alain; Frey-Klett, Pascale
Ectomycorrhizal fungi are surrounded by bacterial communities with which they interact physically and metabolically during their life cycle. These bacteria can have positive or negative effects on the formation and the functioning of ectomycorrhizae. However, relatively little is known about the mechanisms by which ectomycorrhizal fungi and associated bacteria interact. To understand how ectomycorrhizal fungi perceive their biotic environment and the mechanisms supporting interactions between ectomycorrhizal fungi and soil bacteria, we analysed the pairwise transcriptomic responses of the ectomycorrhizal fungus Laccaria bicolor (Basidiomycota: Agaricales) when confronted with beneficial, neutral or detrimental soil bacteria. Comparative analyses of the three transcriptomes indicated that the fungus reacted differently to each bacterial strain. Similarly, each bacterial strain produced a specific and distinct response to the presence of the fungus. Despite these differences in responses observed at the gene level, we found common classes of genes linked to cell-cell interaction, stress response and metabolic processes to be involved in the interaction of the four microorganisms.
Killing of Staphylococcus aureus and Salmonella enteritidis and neutralization of lipopolysaccharide by 17-residue bovine lactoferricins: improved activity of Trp/Ala-containing molecules
Hao, Ya; Yang, Na; Wang, Xiumin; Teng, Da; Mao, Ruoyu; Wang, Xiao; Li, Zhanzhan; Wang, Jianhua
Bovine lactoferricin (LfcinB) has potent antibacterial, antifungal and antiparasitic activities but is also hemolytic. Our objective was to identify LfcinB17-31 derivatives with reduced hemolysis and improved antimicrobial activity via substituting Cys3, Arg4, Gln7, Met10, and Gly14 with more hydrophobic residues. Two peptides, Lfcin4 and Lfcin5, showed higher activity against Staphylococcus aureus and Salmonella enteritidis and lower hemolytic activity than the parent peptide LfcinB17-31. These peptides permeabilized the outer and inner membranes of S. enteritidis; however, Lfcin5 did not permeabilize the inner membrane of S. aureus. Gel retardation and circular dichroism spectra showed that Lfcin4 and Lfcin5 bound to bacterial genomic DNA. Lfcin4 inhibited DNA, RNA and protein synthesis. Both peptides induced the peeling of membranes and the lysis of S. enteritidis. At doses of 10 and 15 mg/kg, Lfcin4 and Lfcin5 reduced the bacterial counts in infected thigh muscles by 0.03‒0.10 and 0.05‒0.63 log10 CFU/g of tissue, respectively, within 10 h. Lfcin4 and Lfcin5 enhanced the survival rate of endotoxemic mice; reduced serum IL-6, IL-1β and TNF-α levels; and protected mice from lipopolysaccharide-induced lung injury. These data suggest that Lfcin4 and Lfcin5 may be antimicrobial and anti-endotoxin peptides that could serve as the basis for the development of dual-function agents. PMID:28287172
Killing of Staphylococcus aureus and Salmonella enteritidis and neutralization of lipopolysaccharide by 17-residue bovine lactoferricins: improved activity of Trp/Ala-containing molecules.
Hao, Ya; Yang, Na; Wang, Xiumin; Teng, Da; Mao, Ruoyu; Wang, Xiao; Li, Zhanzhan; Wang, Jianhua
Bovine lactoferricin (LfcinB) has potent antibacterial, antifungal and antiparasitic activities but is also hemolytic. Our objective was to identify LfcinB17-31 derivatives with reduced hemolysis and improved antimicrobial activity via substituting Cys3, Arg4, Gln7, Met10, and Gly14 with more hydrophobic residues. Two peptides, Lfcin4 and Lfcin5, showed higher activity against Staphylococcus aureus and Salmonella enteritidis and lower hemolytic activity than the parent peptide LfcinB17-31. These peptides permeabilized the outer and inner membranes of S. enteritidis; however, Lfcin5 did not permeabilize the inner membrane of S. aureus. Gel retardation and circular dichroism spectra showed that Lfcin4 and Lfcin5 bound to bacterial genomic DNA. Lfcin4 inhibited DNA, RNA and protein synthesis. Both peptides induced the peeling of membranes and the lysis of S. enteritidis. At doses of 10 and 15 mg/kg, Lfcin4 and Lfcin5 reduced the bacterial counts in infected thigh muscles by 0.03‒0.10 and 0.05‒0.63 log10 CFU/g of tissue, respectively, within 10 h. Lfcin4 and Lfcin5 enhanced the survival rate of endotoxemic mice; reduced serum IL-6, IL-1β and TNF-α levels; and protected mice from lipopolysaccharide-induced lung injury. These data suggest that Lfcin4 and Lfcin5 may be antimicrobial and anti-endotoxin peptides that could serve as the basis for the development of dual-function agents.
Expression of the surface antigen 4F2hc affects system-L-like neutral-amino-acid-transport activity in mammalian cells.
Bröer, S; Bröer, A; Hamprecht, B
Mammalian cells possess a variety of amino acid-transport systems with overlapping substrate specificity. System L is one of the major amino acid-transport systems of non-epithelial cells. By expression cloning we have recently demonstrated that the surface antigen 4F2hc (CD98) is a necessary component for expression of system-L-like amino acid-transport activity in C6-BU-1 rat glioma cells [Bröer, Bröer and Hamprecht (1995) Biochem. J. 312, 863-870]. 4F2hc mRNA was detected in CHO cells, COS cells, activated lymphocytes isolated from mouse spleen and primary cultures of astrocytes. In all these cell types, Na+-independent isoleucine transport was mediated by system L. No contribution of system y+L to isoleucine or arginine transport was detected in C6-BU-1 cells. In lymphocytes, both system-L-like amino acid-transport activity and 4F2hc mRNA levels increased after treatment with phorbol ester plus ionomycin. Antisense oligonucleotides caused modest inhibition of Na+-independent isoleucine transport in C6-BU-1 cells and primary cultures of astroglial cells, whereas arginine transport was unaffected. Overexpression of 4F2hc cDNA in CHO cells resulted in an increase in Na+-independent isoleucine transport. PMID:9182715
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.
Qiao, Xue; Lin, Xiong-hao; Ji, Shuai; Zhang, Zheng-xiang; Bo, Tao; Guo, De-an; Ye, Min
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.
Cross-clade neutralizing antibodies against HIV-1 induced in rabbits by focusing the immune response on a neutralizing epitope
Zolla-Pazner, Susan; Cohen, Sandra; Pinter, Abraham; Krachmarov, Chavdar; Wrin, Terri; Wang Shixia; Lu Shan
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.
Selection of Peptide Mimics of HIV-1 Epitope Recognized by Neutralizing Antibody VRC01
Chikaev, Anton N.; Bakulina, Anastasiya Yu.; Burdick, Ryan C.; Karpenko, Larisa I.; Pathak, Vinay K.; Ilyichev, Alexander A.
The ability to induce anti-HIV-1 antibodies that can neutralize a broad spectrum of viral isolates from different subtypes seems to be a key requirement for development of an effective HIV-1 vaccine. The epitopes recognized by the most potent broadly neutralizing antibodies that have been characterized are largely discontinuous. Mimetics of such conformational epitopes could be potentially used as components of a synthetic immunogen that can elicit neutralizing antibodies. Here we used phage display technology to identify peptide motifs that mimic the epitope recognized by monoclonal antibody VRC01, which is able to neutralize up to 91% of circulating primary isolates. Three rounds of biopanning were performed against 2 different phage peptide libraries for this purpose. The binding specificity of selected phage clones to monoclonal antibody VRC01 was estimated using dot blot analysis. The putative peptide mimics exposed on the surface of selected phages were analyzed for conformational and linear homology to the surface of HIV-1 gp120 fragment using computational analysis. Corresponding peptides were synthesized and checked for their ability to interfere with neutralization activity of VRC01 in a competitive inhibition assay. One of the most common peptides selected from 12-mer phage library was found to partially mimic a CD4-binding loop fragment, whereas none of the circular C7C-mer peptides was able to mimic any HIV-1 domains. However, peptides identified from both the 12-mer and C7C-mer peptide libraries showed rescue of HIV-1 infectivity in the competitive inhibition assay. The identification of epitope mimics may lead to novel immunogens capable of inducing broadly reactive neutralizing antibodies. PMID:25785734
Environmental neutralization of polonium-218
Goldstein, S.D.; Hopke, P.K.
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.
Neutralization of reovirus: the gene responsible for the neutralization antigen
The S1 genome segment of reovirus is linked to type specificity as determined by neutralization antibody. This gene segment codes for a minor outer capsid polypeptide (sigma1). Therefore, sigma1 is the peptide responsible for induction of neutralization antibody and confers type specificity. This biologic property of reovirus was defined using hybrid recombinants clones between reovirus types 1 and 3 and 2 and 3. PMID:925604
Neutralization Serotyping of BK Polyomavirus Infection in Kidney Transplant Recipients
Pastrana, Diana V.; Brennan, Daniel C.; Çuburu, Nicolas; Storch, Gregory A.; Viscidi, Raphael P.; Randhawa, Parmjeet S.; Buck, Christopher B.
BK polyomavirus (BKV or BKPyV) associated nephropathy affects up to 10% of kidney transplant recipients (KTRs). BKV isolates are categorized into four genotypes. It is currently unclear whether the four genotypes are also serotypes. To address this issue, we developed high-throughput serological assays based on antibody-mediated neutralization of BKV genotype I and IV reporter vectors (pseudoviruses). Neutralization-based testing of sera from mice immunized with BKV-I or BKV-IV virus-like particles (VLPs) or sera from naturally infected human subjects revealed that BKV-I specific serum antibodies are poorly neutralizing against BKV-IV and vice versa. The fact that BKV-I and BKV-IV are distinct serotypes was less evident in traditional VLP-based ELISAs. BKV-I and BKV-IV neutralization assays were used to examine BKV type-specific neutralizing antibody responses in KTRs at various time points after transplantation. At study entry, sera from 5% and 49% of KTRs showed no detectable neutralizing activity for BKV-I or BKV-IV neutralization, respectively. By one year after transplantation, all KTRs were neutralization seropositive for BKV-I, and 43% of the initially BKV-IV seronegative subjects showed evidence of acute seroconversion for BKV-IV neutralization. The results suggest a model in which BKV-IV-specific seroconversion reflects a de novo BKV-IV infection in KTRs who initially lack protective antibody responses capable of neutralizing genotype IV BKVs. If this model is correct, it suggests that pre-vaccinating prospective KTRs with a multivalent VLP-based vaccine against all BKV serotypes, or administration of BKV-neutralizing antibodies, might offer protection against graft loss or dysfunction due to BKV associated nephropathy. PMID:22511874
High-Throughput Analysis of Enzyme Activities
Lu, Guoxin
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
Investigation of Relationship between QBO and Ionospheric Neutral Temperature
NASA Astrophysics Data System (ADS)
Saǧır, Selçuk; Atıcı, Ramazan; Özcan, Osman
The relationship between Quasi Biennial Oscillation (QBO) measured at 10 hPa altitude and neutral temperature obtained from NRLMSIS-00 model for 90 km altitude of ionosphere is statistically investigated. For this study, multiple-regression model is used. To see effect on neutral temperature of QBO directions, Dummy variables are added to model established. In the results of performed analysis, it is observed that QBO is effected on neutral temperature of ionosphere. It is determined that 57% of variations at neutral temperature can be explainable by QBO. According to the established model, statistical explainable ratio was determined as 1% that it is the highest rate. Also, it is seen that an increase/a decrease of 1 meter per second at QBO give rise to an increase/a decrease of 0,07 K at neutral temperature.
Presence of neutralizing antibodies to heterologous human immunodeficiency virus type 1 isolates in sera of infected individuals is not predictive of rate of disease progression.
Warren, R Q; Wong, M T; Melcher, G P; Blatt, S P; Zapiola, I; Bouzas, M B; Muchinik, G; Anderson, S A; Kennedy, R C
These studies were undertaken to examine whether the presence of human immunodeficiency virus type 1 (HIV-1)-neutralizing antibodies in sera of infected individuals would alter the rate of disease progression. HIV-1-infected individuals (n = 87) were initially examined for neutralizing activity in vitro against both laboratory and tissue culture-adapted clinical heterologous HIV-1 isolates. The neutralizing activities of sera were determined by a 90% or greater reduction in HIV-1 p24 levels in vitro. In a cross-sectional analysis of all infected individuals, we observed that sera from asymptomatic individuals neutralized a significantly greater number of heterologous HIV-1 isolates than sera from symptomatic patients. Patients who could be followed up longitudinally (n = 24) were then studied to determine the impact of neutralizing antibodies on the rate of disease progression. We observed no significant difference between the numbers of HIV-1 isolates neutralized in vitro by sera from patients who remained clinically stable and by those from patients who progressed rapidly. Our data indicated that the presence or absence of neutralizing antibodies to heterologous HIV-1 isolates was not associated with the rate of disease progression. PMID:7583913
Perturbations to the neutral atmosphere caused by acoustic gravity waves at thermospheric altitudes as obtained from Dynasonde data
NASA Astrophysics Data System (ADS)
Negrea, Catalin; Zabotin, Nikolay; Bullett, Terry
Gravity waves are known to have a major impact on the dynamics of the thermosphere-ionosphere. A number of recent studies addressed the issue of determining the characteristics of thermospheric gravity waves and associated Travelling Ionospheric Disturbances and also their impact on the background system. However, there are currently no methods that would allow for the continuous and uninterrupted study of both the spatial and temporal characteristics of gravity wave activity over a broad range of thermospheric altitudes. We present results obtained using Dynasonde measurements of electron density and ionospheric tilts. The data covers the bottom E- and F-Layers and implicitly contains information on induced perturbations in the horizontal plane at all accessible altitudes. The methodology that we developed is largely automated, allowing for the analysis of large amounts of data. A model of the thermosphere-ionosphere coupling is implemented to infer neutral atmosphere parameters from ionospheric measurements. This is done by accounting for ion-neutral interactions, changes to chemical composition due to wave propagation and the effect of the geomagnetic field. Background neutral temperature, neutral density and neutral composition are used from a numerical model. A sample dataset from October 24th at Wallops Island, Virginia is used to illustrate our approach. The frequency, wavevector components, group velocity, phase speed and amplitude of induced thermospheric and ionospheric perturbations are obtained. These include the TID amplitude as well as the underlying gravity wave amplitude in neutral density, temperature and zonal and meridional winds.
Karyotype Analysis Activity: A Constructivist Learning Design
ERIC Educational Resources Information Center
Ahmed, Noveera T.
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…
A nickel-borate nanoarray: a highly active 3D oxygen-evolving catalyst electrode operating in near-neutral water.
Ji, Xuqiang; Cui, Liang; Liu, Danni; Hao, Shuai; Liu, Jingquan; Qu, Fengli; Ma, Yongjun; Du, Gu; Asiri, Abdullah M; Sun, Xuping
The exploration of high-performance and cost-effective water oxidation catalysts operating under mild conditions is still urgent and challenging. In this communication, a nickel-borate nanoarray supported on carbon cloth (Ni-Bi/CC) has been fabricated through oxidative polarization of a nickel oxide nanoarray on CC (NiO/CC) in a borate electrolyte (pH 9.2). As a 3D electrode, this Ni-Bi/CC exhibits superior catalytic activity for water oxidation in 0.1 M potassium borate (K-Bi) solution, yielding a geometrical catalytic current density of 10 mA cm(-2) at an overpotential of 470 mV. Notably, this electrode also demonstrates outstanding long-term electrochemical durability for 25 h with 100% Faradaic efficiency.
Communication: Classical threshold law for ion-neutral-neutral three-body recombination
Pérez-Ríos, Jesús; Greene, Chris H.
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.
The Relationship between Neutralization Techniques and Induced Abortion
Kalateh Sadati, Ahmad; Tabei, Seyed Ziaaddin; Salehzadeh, Hamzeh; Rahnavard, Farnaz; Namavar Jahromi, Bahia; Hemmati, Soroor
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
Incomplete Neutralization and Deviation from Sigmoidal Neutralization Curves for HIV Broadly Neutralizing Monoclonal Antibodies.
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
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.
Initiation of immune tolerance–controlled HIV gp41 neutralizing B cell lineages
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.
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
Activity Analysis for Cognitive-Perceptual-Motor Dysfunction
ERIC Educational Resources Information Center
Llorens, Lela A.
This paper presents a review of several approaches to activity and task analysis for their selection for use in occupational therapy and proposes a neuro-behavioral approach to activity analysis and selection for use in treatment of cognitive-perceptual-motor dysfunction. (Editors/JA)
Overview of MSFC's Applied Fluid Dynamics Analysis Group Activities
NASA Technical Reports Server (NTRS)
Garcia, Roberto; Griffin, Lisa; Williams, Robert
TD64, the Applied Fluid Dynamics Analysis Group, is one of several groups with high-fidelity fluids design and analysis expertise in the Space Transportation Directorate at Marshall Space Flight Center (MSFC). TD64 assists personnel working on other programs. The group participates in projects in the following areas: turbomachinery activities, nozzle activities, combustion devices, and the Columbia accident investigation.
Sono-incorporation of CuO nanoparticles on the surface and into the mesoporous hexatitanate layers: Enhanced Fenton-like activity in degradation of orange-G at its neutral pH
NASA Astrophysics Data System (ADS)
Sehati, S.; Entezari, M. H.
In this work, for the first time, CuO/Ti6O13 mesoporous nanocomposite was synthesized by direct intercalation of CuO nanoparticles into hexatitanate layers in the presence of ultrasound. In fact mesoporous potassium hexatitanate with high pore size (44.94 nm) was used as a support. CuO nanoparticles were grown into the titanate pores which caused formation of ultra small CuO with uniform size and high surface area. In fact, titanate is considered as a substrate for better dispersion and nucleation of the CuO nanoparticles which prevented the agglomeration and overgrowth of guest molecules. The prepared sample was characterized by XRD, FE-SEM, TEM, UV-vis spectra, N2 adsorption-desorption, Raman spectra and FT-IR techniques. The product was used as a heterogeneous Fenton-like catalyst for the degradation of Orang G (OG). The effect of important parameters, including pH, H2O2 addition rate and catalyst loading on the decolorization of OG were investigated. Based on the results, CuO/Ti6O13 catalyst exhibited high catalytic activity for OG degradation in aqueous solution at neutral pH of the dye. Moreover, breaking of H2O2 during the catalytic reaction was monitored by spectroscopic method. The results confirmed the decomposition of H2O2 to produce rad OH which is the main active species for the degradation of OG.
A shell-neutral modeling approach yields sustainable oyster harvest estimates: a retrospective analysis of the Louisiana state primary seed grounds
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
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.
Linking Temporal Changes in Bacterial Community Structures with the Detection and Phylogenetic Analysis of Neutral Metalloprotease Genes in the Sediments of a Hypereutrophic Lake
Tsuboi, Shun; Yamamura, Shigeki; Imai, Akio; Satou, Takayuki; Iwasaki, Kazuhiro
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
Vendor neutral archive in PACS.
Agarwal, Tapesh Kumar; Sanjeev
An archive is a location containing a collection of records, documents, or other materials of historical importance. An integral part of Picture Archiving and Communication System (PACS) is archiving. When a hospital needs to migrate a PACS vendor, the complete earlier data need to be migrated in the format of the newly procured PACS. It is both time and money consuming. To address this issue, the new concept of vendor neutral archive (VNA) has emerged. A VNA simply decouples the PACS and workstations at the archival layer. This is achieved by developing an application engine that receives, integrates, and transmits the data using the different syntax of a Digital Imaging and Communication in Medicine (DICOM) format. Transferring the data belonging to the old PACS to a new one is performed by a process called migration of data. In VNA, a number of different data migration techniques are available to facilitate transfer from the old PACS to the new one, the choice depending on the speed of migration and the importance of data. The techniques include simple DICOM migration, prefetch-based DICOM migration, medium migration, and the expensive non-DICOM migration. "Vendor neutral" may not be a suitable term, and "architecture neutral," "PACS neutral," "content neutral," or "third-party neutral" are probably better and preferred terms. Notwithstanding this, the VNA acronym has come to stay in both the medical IT user terminology and in vendor nomenclature, and radiologists need to be aware of its impact in PACS across the globe.
Breath gas metabolites and bacterial metagenomes from cystic fibrosis airways indicate active pH neutral 2,3-butanedione fermentation.
Whiteson, Katrine L; Meinardi, Simone; Lim, Yan Wei; Schmieder, Robert; Maughan, Heather; Quinn, Robert; Blake, Donald R; Conrad, Douglas; Rohwer, Forest
The airways of cystic fibrosis (CF) patients are chronically colonized by patient-specific polymicrobial communities. The conditions and nutrients available in CF lungs affect the physiology and composition of the colonizing microbes. Recent work in bioreactors has shown that the fermentation product 2,3-butanediol mediates cross-feeding between some fermenting bacteria and Pseudomonas aeruginosa, and that this mechanism increases bacterial current production. To examine bacterial fermentation in the respiratory tract, breath gas metabolites were measured and several metagenomes were sequenced from CF and non-CF volunteers. 2,3-butanedione was produced in nearly all respiratory tracts. Elevated levels in one patient decreased during antibiotic treatment, and breath concentrations varied between CF patients at the same time point. Some patients had high enough levels of 2,3-butanedione to irreversibly damage lung tissue. Antibiotic therapy likely dictates the activities of 2,3-butanedione-producing microbes, which suggests a need for further study with larger sample size. Sputum microbiomes were dominated by P. aeruginosa, Streptococcus spp. and Rothia mucilaginosa, and revealed the potential for 2,3-butanedione biosynthesis. Genes encoding 2,3-butanedione biosynthesis were disproportionately abundant in Streptococcus spp, whereas genes for consumption of butanedione pathway products were encoded by P. aeruginosa and R. mucilaginosa. We propose a model where low oxygen conditions in CF lung lead to fermentation and a decrease in pH, triggering 2,3-butanedione fermentation to avoid lethal acidification. We hypothesize that this may also increase phenazine production by P. aeruginosa, increasing reactive oxygen species and providing additional electron acceptors to CF microbes.
Anti-cancer Activity of Novel TM4SF5-Targeting Antibodies through TM4SF5 Neutralization and Immune Cell-Mediated Cytotoxicity
Ahn, Hye-Mi; Ryu, Jihye; Song, Jin Myeong; Lee, Yunhee; Kim, Hye-Jin; Ko, Dongjoon; Choi, Inpyo; Kim, Sang Jick; Lee, Jung Weon; Kim, Semi
The transmembrane four L6 family member 5 (TM4SF5) protein is a novel molecular target for the prevention and treatment of hepatocellular carcinoma. TM4SF5 is highly expressed in liver, colon, esophageal, and pancreatic cancers and is implicated in tumor progression. Here, we screened monoclonal antibodies that specifically bound to the extracellular loop 2 (EC2) of TM4SF5 from a phage-displayed murine antibody (single-chain variable fragment; scFv) library. We constructed and characterized chimeric antibodies, Ab27 and Ab79, of scFv fused with Fc domain of human IgG1. The affinity (KD) of Ab27 and Ab79 for soluble EC2 was approximately 9.2 nM and 16.9 nM, respectively, as determined by surface plasmon resonance analysis. Ab27 and Ab79 efficiently bound to native TM4SF5 on the cell surface were internalized into the cancer cells, leading to a decrease in cell surface TM4SF5. Ab27 and Ab79 inhibited the proliferation and invasion of TM4SF5-positive liver and colon cancer cells and reduced FAK and c-Src phosphorylation. Ab27 and Ab79 also enhanced anoikis sensitivity and reduced survivin. Ab27 mediated antibody-dependent cell-mediated cytotoxicity in vitro. Ab27 and Ab79 efficiently inhibited tumor growth in a liver cancer xenograft model. These results strongly support the further development of Ab27 as a novel anti-cancer agent in the clinic. PMID:28255353
Measurement of the relative amplitude and strong phase between antineutral D meson decaying to kaon+ resonance kaon- and neutral D meson decaying to kaon+ resonance kaon- via Dalitz plot analysis of neutral D meson decaying to kaon+ kaon- neutral pion decays
NASA Astrophysics Data System (ADS)
Naik, Paras P.
I present physics concepts, which are useful to understand our analyses, and describe the CLEO III and CLEO-c experiments at the Cornell Electron Storage Ring. I also present motivations for a Dalitz plot analysis of the Cabibbo-suppressed charmed meson decay mode D0 → K+K-pi 0 at CLEO. The analysis uses 9.0 fb-1 of data collected at s ≈ 10.58 GeV with the CLEO III detector. We find the strong phase difference deltaD ≡ argAD 0→K*+K- AD0→K*+K - = 332° +/- 8° +/- 11° and relative amplitude rD ≡ AD0→ K*+K-A D0→K*+K- = 0.52 +/- 0.05 +/- 0.04. This measurement indicates significant destructive interference between D0 → K+(K-pi 0)K*- and D0 → K- (K+pi0) K*+ in the D 0 → K+ K-pi0 Dalitz plot region where these two modes overlap. The fit includes the K*+/- and φ resonances and a non-resonant amplitude, and the measured fit fractions for each resonance (with statistical uncertainty only) are (46.1 +/- 3.1)% for the K*+, (12.3 +/- 2.2)% for the K*-, (14.9 +/- 1.6)% for the φ, and (36.0 +/- 3.7)% for the non-resonant contribution. We find deltaD = 313° +/- 9° (stat.) and an amplitude ratio of rD = 0.52 +/- 0.05 (stat.) from a second fit which substitutes scalar kappa+/- (mass 878 MeV/c2, width 499 MeV/c2) amplitudes for the non-resonant amplitude. The measured fit fractions for each resonance (with statistical uncertainty only) are (48.1 +/- 4.5)% for the K*+, (12.9 +/- 2.6)% for the K*-, (16.1 +/- 1.9)% for the φ, (12.6 +/- 5.8)% for the kappa+, and (11.1 +/- 4.7)% for the kappa-. We also investigate the D 0 → K+K -pi0 Dalitz plot in 281 pb-1 of data collected at s ≈ 3.77 GeV with the CLEO-c detector. We find results which are consistent with the CLEO III analysis. I conclude by summarizing our results and present a brief appendix detailing the K-matrix formalism.
Activity Landscape Plotter: A Web-Based Application for the Analysis of Structure-Activity Relationships.
González-Medina, Mariana; Méndez-Lucio, Oscar; Medina-Franco, José L
Activity landscape modeling is a powerful method for the quantitative analysis of structure-activity relationships. This cheminformatics area is in continuous growth, and several quantitative and visual approaches are constantly being developed. However, these approaches often fall into disuse due to their limited access. Herein, we present Activity Landscape Plotter as the first freely available web-based tool to automatically analyze structure-activity relationships of compound data sets. Based on the concept of activity landscape modeling, the online service performs pairwise structure and activity relationships from an input data set supplied by the user. For visual analysis, Activity Landscape Plotter generates Structure-Activity Similarity and Dual-Activity Difference maps. The user can interactively navigate through the maps and export all the pairwise structure-activity information as comma delimited files. Activity Landscape Plotter is freely accessible at https://unam-shiny-difacquim.shinyapps.io/ActLSmaps /.
Overview of MSFC's Applied Fluid Dynamics Analysis Group Activities
NASA Technical Reports Server (NTRS)
Garcia, Roberto; Griffin, Lisa; Williams, Robert
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.
Neutron Activation Analysis, A Titanium Material Study
NASA Astrophysics Data System (ADS)
Dresser, Charles
In order to obtain faster and more accurate measurements of radioactive contaminates within a sample of titanium we expose it to a neutron flux. This flux will activate the stable and quasi stable (those with extremely long half lives) isotopes into resultant daughter cells that are unstable which will result in shorter half lives on the order of minutes to days. We measured the resulting decays in the Germanium Crystal Detector and obtained a complex gamma spectrum. A mathematical model was used to recreate the production of the measured isotopes in the neutron flux and the resultant decays. Using this model we calculated the mass percent of the contaminate isotopes inside our titanium sample. Our mathematical model accounted for two types of neutron activation, fast or thermal activation, since this would determine which contaminate was the source of our signals. By looking at the percent abundances, neutron absorption cross-sections and the resulting mass percents of each contaminate we are able to determine the exact source of our measured signals. Additionally we implemented a unique ratio method to cross check the mathematical model. Our results have verified that for fast neutron activation and thermal neutron activation the method is accurate.
Low-energy neutral-atom spectrometer
Voss, D.E.; Cohen, S.A.
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.
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…
Method for photon activation positron annihilation analysis
Akers, Douglas W.
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.
Neutral Supersymmetric Higgs Boson Searches
Robinson, Stephen Luke
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
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
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
Catalytic Transfer of Magnetism using a Neutral Iridium Phenoxide Complex
Ruddlesden, Amy J.; Mewis, Ryan E.; Green, Gary G. R.; Whitwood, Adrian C.; Duckett, Simon B.
A novel neutral iridium carbene complex Ir(κC,O-L1)(COD) (1) [where COD = cyclooctadiene and L1 = 3-(2-methylene-4-nitrophenolate)-1-(2,4,6-trimethylphenyl) imidazolylidene] with a pendant alkoxide ligand has been prepared and characterized. It contains a strong Ir-O bond and X-ray analysis reveals a distorted square planar structure. NMR spectroscopy reveals dynamic solution state behavior commensurate with rapid seven-membered ring flipping. In CD2Cl2 solution, under hydrogen at low temperature, this complex dominates although it exists in equilibrium with a reactive iridium dihydride cyclooctadiene complex. 1 reacts with pyridine and H2 to form neutral Ir(H)2(κC,O-L1)(py)2 which also exists in two conformers that differ according to the orientation of the seven-membered metallocycle and whilst its Ir-O bond remains intact, the complex undergoes both pyridine and H2 exchange. As a consequence, when placed under parahydrogen, efficient polarization transfer catalysis (PTC) is observed via the Signal Amplification By Reversible Exchange (SABRE) approach. Due to the neutral character of this catalyst, good hyperpolarization activity is shown in a wide range of solvents for a number of substrates. These observations reflect a dramatic improvement in solvent tolerance of SABRE over that reported for the best PTC precursor IrCl(IMes)(COD). For THF, the associated 1H NMR signal enhancement for the ortho proton signal of pyridine shows an increase of 600-fold at 298 K. The level of signal enhancement can be increased further through warming or varying the magnetic field experienced by the sample at the point of catalytic magnetization transfer.
Surface conversion techniques for low energy neutral atom imagers
NASA Technical Reports Server (NTRS)
Quinn, J. M.
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.
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.…
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...
Digital image processing and analysis for activated sludge wastewater treatment.
Khan, Muhammad Burhan; Lee, Xue Yong; Nisar, Humaira; Ng, Choon Aun; Yeap, Kim Ho; Malik, Aamir Saeed
Activated sludge system is generally used in wastewater treatment plants for processing domestic influent. Conventionally the activated sludge wastewater treatment is monitored by measuring physico-chemical parameters like total suspended solids (TSSol), sludge volume index (SVI) and chemical oxygen demand (COD) etc. For the measurement, tests are conducted in the laboratory, which take many hours to give the final measurement. Digital image processing and analysis offers a better alternative not only to monitor and characterize the current state of activated sludge but also to predict the future state. The characterization by image processing and analysis is done by correlating the time evolution of parameters extracted by image analysis of floc and filaments with the physico-chemical parameters. This chapter briefly reviews the activated sludge wastewater treatment; and, procedures of image acquisition, preprocessing, segmentation and analysis in the specific context of activated sludge wastewater treatment. In the latter part additional procedures like z-stacking, image stitching are introduced for wastewater image preprocessing, which are not previously used in the context of activated sludge. Different preprocessing and segmentation techniques are proposed, along with the survey of imaging procedures reported in the literature. Finally the image analysis based morphological parameters and correlation of the parameters with regard to monitoring and prediction of activated sludge are discussed. Hence it is observed that image analysis can play a very useful role in the monitoring of activated sludge wastewater treatment plants.
Modeling of modification experiments involving neutral-gas release
Bernhardt, P.A.
Many experiments involve the injection of neutral gases into the upper atmosphere. Examples are critical velocity experiments, MHD wave generation, ionospheric hole production, plasma striation formation, and ion tracing. Many of these experiments are discussed in other sessions of the Active Experiments Conference. This paper limits its discussion to: (1) the modeling of the neutral gas dynamics after injection, (2) subsequent formation of ionosphere holes, and (3) use of such holes as experimental tools.
Characterization of Hierarchical α-MoO3 Plates Toward Resistive Heating Synthesis: Electrochemical Activity of α-MoO3/Pt Modified Electrode Toward Methanol Oxidation in Neutral pH.
Filippo, Emanuela; Baldassarre, Francesca; Tepore, Marco; Guascito, Maria Rachele; Chirizzi, Daniela; Tepore, Antonio
The growth of MoO3 hierarchical plates was obtained by direct resistive heating of molybdenum foil at ambient pressure in absence of any catalysts and templates. Plates synthesized after 60 min resistive heating typically growth in an single-crystalline orthorhombic structure that develop preferentially in [001] direction, as characterized by HRTEM, SAD and Raman-scattering measurements. They are about 100-200nm in thickness and a few tens micrometers in length. As heating time proceeds to 80 min, plates of α-MoO3 form a branched structure. A more attentive look shows that a primary plates formed at until 60 min could serve as substrates for the subsequent growth of secondary belts. Moreover, a full electrochemical characterization of α-MoO3 plates on platinum electrodes was done by Cyclic Voltammetric experiments, at pH 7 in phosphate buffer, to probe the activity of the proposed composite material as anode to methanol electrooxidation. Reported results indicate that Pt MoO3 modified electrodes are appropriate to develop new amperometric non-enzymatic sensor for methanol measurements and as anode in Direct Methanol Fuel Cells (DMFCs) making at neutral pH.
Gas cell neutralizers (Fundamental principles)
Fuehrer, B.
Neutralizing an ion-beam of the size and energy levels involved in the neutral-particle-beam program represents a considerable extension of the state-of-the-art of neutralizer technology. Many different mediums (e.g., solid, liquid, gas, plasma, photons) can be used to strip the hydrogen ion of its extra electron. A large, multidisciplinary R and D effort will no doubt be required to sort out all of the ''pros and cons'' of these various techniques. The purpose of this particular presentation is to discuss some basic configurations and fundamental principles of the gas type of neutralizer cell. Particular emphasis is placed on the ''Gasdynamic Free-Jet'' neutralizer since this configuration has the potential of being much shorter than other type of gas cells (in the beam direction) and it could operate in nearly a continuous mode (CW) if necessary. These were important considerations in the ATSU design which is discussed in some detail in the second presentation entitled ''ATSU Point Design''.
Reduced neutral XLPE cable design
Valli, G.F.; Zawadzki, J.A.; Orton, H.E. )
This paper describes the theoretical, laboratory and economic analyses undertaken to determine the optimum metallic concentric neutral design for its single conductor 750 and 500 kcmil aluminum XLPE 15 kV insulated concentric-neutral type feeder cables. The results suggest that reducing the cross-sectional area of this concentric neutral from the currently-recognized industry standard of 20 percent of the central conductor to 7% results in overall present-worth system cost saving of approximately $3 per conductor meter or approximately 22% of the cable first cost. The neutral configuration ultimately chosen to replace the previous standard 37 - number 14 AWG wires was 2 - 1 inch {times} 5 mil tinned copper tapes overlapped by 25%. Line voltage fault test were run in the high-power laboratory on samples with various neutral configurations to confirm they would successfully pass our worst-case fault duty of 10 kA for 20 cycles (i.e., .33 sec) with no reclosing.
ACTIVE: A Tool for Integrating Analysis Contracts
for modeling systems in the Architecture Analysis and Design Language (AADL). In the paper we analyze the problems that occur when independently...5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) Dionisio de Niz Ivan Ruchkin; Sagar Chaki; David Garlan 5d. PROJECT NUMBER 5e. TASK NUMBER 5f. WORK UNIT...tectural description language because AADL offers a con- venient way to represent the structural, design-time aspect of the system. In particular
Potent neutralizing monoclonal antibodies against Ebola virus infection
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
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
Neutral recirculation—the key to control of divertor operation
NASA Astrophysics Data System (ADS)
Kukushkin, A. S.; Pacher, H. D.
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.
Neutral current pi0 production in MiniBooNE
Link, J.M.; /Virginia Tech.
This paper describes the analysis used to determine the neutral current {pi}{sup 0} production in MiniBooNE in bins of momentum. Additionally, a measurement of the relative coherent production of {pi}{sup 0}s is discussed. The coherent production rate is found to be (19.5 {+-}1.1 (stat) {+-}2.5 (sys))% of the total exclusive neutral current {pi}{sup 0} production rate.
Neutralization of H- at Nanostructured Surfaces
NASA Astrophysics Data System (ADS)
Obreshkov, Boyan; Thumm, Uwe
The charge transfer rates and the neutralization probabilities for hydrogen anions colliding with nanostructured (vicinal) surfaces are obtained by direct numerical integration of the time-dependent Schroedinger equation for the motion of the active electron in the field of the projectile-surface compound. The electronic structure of the surface is calculated from a Thomas-Fermi - von Weizsaecker statistical model with local density approximation for the exchange-correlation energy. In fixed-ion approximation, the decay rate of the electronic state of the anion in front of the surface is obtained by projecting the density of states of the collision system onto the unperturbed projectile level. The ion neutralization probability is calculated from this static width within a rate equation approach for a set of broken-straight-line collision trajectories for kinetic energies of 1 keV. The dependence of decay rates and neutralization probabilities on the surface morphology and the scattering trajectories, and a comparison of our numerical results with the experiments will be discussed.
Active vision in satellite scene analysis
NASA Technical Reports Server (NTRS)
Naillon, Martine
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.
Simulations of neutralized final focus
Welch, D.R.; Rose, D.V.; Genoni, T.C.; Yu, S.S.; Barnard, J.J.
In order to drive an inertial fusion target or study high energy density physics with heavy ion beams, the beam radius must be focused to < 3 mm and the pulse length must be compressed to < 10 ns. The conventional scheme for temporal pulse compression makes use of an increasing ion velocity to compress the beam as it drifts and beam space charge to stagnate the compression before final focus. Beam compression in a neutralizing plasma does not require stagnation of the compression, enabling a more robust method. The final pulse shape at the target can be programmed by an applied velocity tilt. In this paper, neutralized drift compression is investigated. The sensitivity of the compression and focusing to beam momentum spread, plasma, and magnetic field conditions is studied with realistic driver examples. Using the 3D particle-in-cell code, we examine issues associated with self-field generation, stability, and vacuum-neutralized transport transition and focusing.
Higher Throughput Quantification of Neutralizing Antibody to Herpes Simplex Viruses.
Blevins, Tamara P; Mitchell, Michelle C; Korom, Maria; Wang, Hong; Yu, Yinyi; Morrison, Lynda A; Belshe, Robert B
We report a rapid, higher throughput method for measuring neutralizing antibody to herpes simplex virus (HSV) in human sera. Clinical isolates and sera from the Herpevac Trial for Women were used in a colorimetric assay in which infection of tissue culture (lack of neutralization) was indicated by substrate metabolism by beta-galactosidase induced in the ELVIS cell line. The neutralization assay was optimized by addition of guinea pig complement, which particularly enhanced neutralizing antibody titers to HSV-2. Higher neutralizing antibody titers were also achieved using virus particles isolated from the supernatant of infected cells rather than lysate of infected cells as the source of virus. The effect of assay incubation time and incubation time with substrate were also optimized. We found that incubating with substrate until a standard optical density of 1.0 was reached permitted a better comparison among virus isolates, and achieved reliable measurement of neutralizing antibody activity. Interestingly, in contrast to results in the absence of complement, addition of complement allowed sera from HSV-2 gD-vaccinated subjects to neutralize HSV-1 and HSV-2 clinical and laboratory isolates with equal potency.
Measurements of the E region neutral wind field
NASA Technical Reports Server (NTRS)
Cogger, L. L.; Murphree, J. S.; Tepley, C. A.; Meriwether, J. W., Jr.
The neutral E-region wind field was measured at Calgary, Canada (51 N, 114 N) during 75 nights in 1982. Observations of the Doppler shift of the 5577-A emission line of atomic oxygen using a Fabry-Perot interferometer were converted to horizontal wind vectors. From the analysis of the data, four categories of wind characteristics were identified. In order of increasing magnetic activity these categories are: (1) wind field mostly variable in space and time; (2) predominantly equatorward flow throughout the night, (3) predominantly poleward flow throughout the night and (4) north-westward flow before midnight and southward after midnight. The wind magnitude was also variable and on some disturbed days exceeded 200 m/s.
A Re-Examiniation of Phonological Neutralization.
ERIC Educational Resources Information Center
Dinnsen, D.
Reviews research studies that raise serious questions about phonological neutralization, that is, the merger of a contrast in certain contexts. Some findings cast doubt on the very existence of neutralization and the correctness of the theoretical principles that make assumptions based on neutralization. Reanalyzes neutralization in light of these…
Passive versus active mitigation cost analysis
Parazin, R.J.; Galbraith, J.D.
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.
The neutral surface layer above rough surfaces
NASA Astrophysics Data System (ADS)
Smedman, Ann-Sofi; Sahlee, Erik
It is generally accepted that turbulent fluxes (momentum and scalar fluxes) are approx. constant with height above horizontal surfaces with low roughness. But what will happen when the roughness sub-layer is large as found over cities, forests and rough seas? In a study of the kinematic structure of the near neutral atmospheric surface layer, Högström, Hunt and Smedman, 2002, it was demonstrated that a model with detached eddies from above the surface layer impinging on to the surface (Hunt and Morison, 2000) could explain some of the observed features in the neutral atmospheric boundary layer. Thus the detached eddy model proved successful in explaining the dynamic structure of the near neutral atmospheric surface layer, especially the shape of the spectra of the wind components and scalars and corresponding fluxes. Here we make the hypothesis that the detached-eddy model can also be used to explain the experimental results related to the 3-dimensional turbulence structure above rough surfaces. Measurements are taken both over land (grass and forest) and over sea (Baltic Sea and hurricane Fabian in the Atlantic) above the roughness sub-layer. Analysis of the turbulence structure shows a striking similarity between the different sites. Hunt, J.C.R and Morrison, J.F., 2000: Eddy structure in turbulent boundary layers, Euro. J. Mech. B-Fluids, 19, 673-694. Högström, U., Hunt, J.C.R., and Smedman, A., 2002: Theory and measurements for turbulence spectra and variances in the atmospheric neutral surface layer, Bound.-Layer Meteorol., 103,101-124.
PEG conjugation moderately protects adeno-associated viral vectors against antibody neutralization.
Lee, Gary K; Maheshri, Narendra; Kaspar, Brian; Schaffer, David V
AAV gene therapy vectors have significant clinical promise, but serum neutralization poses a challenge that must be overcome. We have examined the potential of conjugating the AAV surface with activated polyethylene glycol chains to protect the vector from neutralizing antibodies. Two key parameters were investigated: the polymer chain size and the PEG:lysine conjugation ratio. Transduction data revealed that the vector is fully infectious until a critical PEG conjugation reaction ratio was exceeded, and this critical level was found to vary with polymer chain size. At this key conjugation ratio, however, particles were moderately protected from serum neutralization, 2.3-fold over unmodified vector, demonstrating that there is a small window of PEGylation for which particles are still fully infective and benefit from antibody protection. TEM results and structural analysis indicate that the drop of infectivity as the PEG concentration is increased beyond the critical conjugation ratio may be due to a combination of steric interference with viral regions necessary for infection as well as reaction at important lysine residues. However, this first study analyzing the potential of PEG to protect AAV from serum neutralization shows that the approach has promise, which can be further enhanced if the locations of PEG attachment can be more finely controlled.
Instrumental Neutron Activation Analysis and Multivariate Statistics for Pottery Provenance
NASA Astrophysics Data System (ADS)
Glascock, M. D.; Neff, H.; Vaughn, K. J.
The application of instrumental neutron activation analysis and multivariate statistics to archaeological studies of ceramics and clays is described. A small pottery data set from the Nasca culture in southern Peru is presented for illustration.
Neutron-activation analysis applied to copper ores and artifacts
NASA Technical Reports Server (NTRS)
Linder, N. F.
Neutron activation analysis is used for quantitative identification of trace metals in copper. Establishing a unique fingerprint of impurities in Michigan copper would enable identification of artifacts made from this copper.
Phosphorus Determination by Derivative Activation Analysis: A Multifaceted Radiochemical Application.
ERIC Educational Resources Information Center
Kleppinger, E. W.; And Others
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)
Elemental analysis of combustion products by neutron activation
Heft, R.E.; Koszykowski, R.F.
This paper gives a brief description of the neutron activation analysis method, which is being used to determine the elemental profile of combustion products from coal-fired power plants, oil shale retorting, and underground coal gasification. (DLC)
Activation analysis of the compact ignition tokamak
Selcow, E.C.
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.
Swimming active droplet: A theoretical analysis
NASA Astrophysics Data System (ADS)
Schmitt, M.; Stark, H.
Recently, an active microswimmer was constructed where a micron-sized droplet of bromine water was placed into a surfactant-laden oil phase. Due to a bromination reaction of the surfactant at the interface, the surface tension locally increases and becomes non-uniform. This drives a Marangoni flow which propels the squirming droplet forward. We develop a diffusion-advection-reaction equation for the order parameter of the surfactant mixture at the droplet interface using a mixing free energy. Numerical solutions reveal a stable swimming regime above a critical Marangoni number M but also stopping and oscillating states when M is increased further. The swimming droplet is identified as a pusher whereas in the oscillating state it oscillates between being a puller and a pusher.
Structural basis of hepatitis C virus neutralization by broadly neutralizing antibody HCV1
Kong, Leopold; Giang, Erick; Robbins, Justin B.; Stanfield, Robyn L.; Burton, Dennis R.; Wilson, Ian A.; Law, Mansun
Hepatitis C virus (HCV) infects more than 2% of the global population and is a leading cause of liver cirrhosis, hepatocellular carcinoma, and end-stage liver diseases. Circulating HCV is genetically diverse, and therefore a broadly effective vaccine must target conserved T- and B-cell epitopes of the virus. Human mAb HCV1 has broad neutralizing activity against HCV isolates from at least four major genotypes and protects in the chimpanzee model from primary HCV challenge. The antibody targets a conserved antigenic site (residues 412-423) on the virus E2 envelope glycoprotein. Two crystal structures of HCV1 Fab in complex with an epitope peptide at 1.8-{angstrom} resolution reveal that the epitope is a {beta}-hairpin displaying a hydrophilic face and a hydrophobic face on opposing sides of the hairpin. The antibody predominantly interacts with E2 residues Leu{sup 413} and Trp{sup 420} on the hydrophobic face of the epitope, thus providing an explanation for how HCV isolates bearing mutations at Asn{sup 415} on the same binding face escape neutralization by this antibody. The results provide structural information for a neutralizing epitope on the HCV E2 glycoprotein and should help guide rational design of HCV immunogens to elicit similar broadly neutralizing antibodies through vaccination.
Structural basis of anthrax edema factor neutralization by a neutralizing antibody
Makiya, Michelle; Dolan, Michael; Agulto, Liane; Purcell, Robert; Chen, Zhaochun
Fine epitope mapping of EF13D, ahighly potent neutralizing monoclonal antibody specific for the anthrax edema factor (EF), was accomplished through random mutagenesis and yeast surface display. A yeast-displayed library of single point mutants of an EF domain III (DIII), comprising amino acids624-800, was constructed by random mutagenesis and screened for reduced binding to EF13D. With this method, residues Leu 667, Ser 668, Arg 671, and Arg 672 were identified as key residues important for EF13D binding. They form a contiguous patch on a solvent-exposed surface at one end of the four-helixbundle of DIII. Computational protein-protein docking experiments between an EF13D model and a crystal structure of EF indicate that the EF13D heavy chain complementarity-determining region 3 (HCDR3) is deeply buried within a hydrophobic cleft between two helices of DIII and interacts directly with residues Leu 667, Ser 668, Arg 671 and Arg 672, providing an explanation for the high binding affinity. In addition, they show that the HCDR3 binding site overlaps with the binding site of the N-terminal lobe of calmodulin (CaM), an EF enzymatic activator, consistent with a previous finding showing direct competition with CaM that results in neutralization of EF. Identifying the neutralization epitope of EF13D on EF improves our understanding of the neutralization mechanism and has implications for vaccine development. PMID:22155239
Broadly neutralizing antibodies developed by an HIV-positive elite neutralizer exact a replication fitness cost on the contemporaneous virus.
Sather, D Noah; Carbonetti, Sara; Kehayia, Jenny; Kraft, Zane; Mikell, Iliyana; Scheid, Johannes F; Klein, Florian; Stamatatos, Leonidas
Approximately 1% of those infected with HIV-1 develop broad and potent serum cross-neutralizing antibody activities. It is unknown whether or not the development of such immune responses affects the replication of the contemporaneous autologous virus. Here, we defined a pathway of autologous viral escape from contemporaneous potent and broad serum neutralizing antibodies developed by an elite HIV-1-positive (HIV-1(+)) neutralizer. These antibodies potently neutralize diverse isolates from different clades and target primarily the CD4-binding site (CD4-BS) of the viral envelope glycoprotein. Viral escape required mutations in the viral envelope glycoprotein which limited the accessibility of the CD4-binding site to the autologous broadly neutralizing anti-CD4-BS antibodies but which allowed the virus to infect cells by utilizing CD4 receptors on their surface. The acquisition of neutralization resistance, however, resulted in reduced cell entry potential and slower viral replication kinetics. Our results indicate that in vivo escape from autologous broadly neutralizing antibodies exacts fitness costs to HIV-1.
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.
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.
N(1520) 3/2- Helicity Amplitudes from an Energy-Independent Multipole Analysis Based on New Polarization Data on Photoproduction of Neutral Pions
NASA Astrophysics Data System (ADS)
Hartmann, J.; Dutz, H.; Anisovich, A. V.; Bayadilov, D.; Beck, R.; Becker, M.; Beloglazov, Y.; Berlin, A.; Bichow, M.; Böse, S.; Brinkmann, K.-Th.; Crede, V.; Dieterle, M.; Eberhardt, H.; Elsner, D.; Fornet-Ponse, K.; Friedrich, St.; Frommberger, F.; Funke, Ch.; Gottschall, M.; Gridnev, A.; Grüner, M.; Gutz, E.; Hammann, Ch.; Hannappel, J.; Hannen, V.; Herick, J.; Hillert, W.; Hoffmeister, Ph.; Honisch, Ch.; Jahn, O.; Jude, T.; Käser, A.; Kaiser, D.; Kalinowsky, H.; Kalischewski, F.; Klassen, P.; Keshelashvili, I.; Klein, F.; Klempt, E.; Koop, K.; Krusche, B.; Kube, M.; Lang, M.; Lopatin, I.; Makonyi, K.; Messi, F.; Metag, V.; Meyer, W.; Müller, J.; Nanova, M.; Nikonov, V.; Novinski, D.; Novotny, R.; Piontek, D.; Rosenbaum, C.; Roth, B.; Reicherz, G.; Rostomyan, T.; Sarantsev, A.; Schmidt, Ch.; Schmieden, H.; Schmitz, R.; Seifen, T.; Sokhoyan, V.; Thämer, Ph.; Thiel, A.; Thoma, U.; Urban, M.; van Pee, H.; Walther, D.; Wendel, Ch.; Wiedner, U.; Wilson, A.; Winnebeck, A.; Witthauer, L.; Wunderlich, Y.; Cbelsa/Taps Collaboration
New data on the polarization observables T, P, and H for the reaction γp→pπ0 are reported. The results are extracted from azimuthal asymmetries when a transversely polarized butanol target and a linearly polarized photon beam are used. The data were taken at the Bonn electron stretcher accelerator ELSA using the CBELSA/TAPS detector. These and earlier data are used to perform a truncated energy-independent partial wave analysis in sliced-energy bins. This energy-independent analysis is compared to the results from energy-dependent partial wave analyses.
Analysis of Smad Phosphatase Activity In Vitro.
Shen, Tao; Qin, Lan; Lin, Xia
Phosphorylation of Smad1/5/8 at the C-terminal SXS motif by BMP type I receptors is one of the most critical events in BMP signaling. Conversely, protein phosphatases that dephosphorylate phospho-Smad1/5/8 can consequently prevent or terminate BMP signaling. PPM1H is an undercharacterized phosphatase in the PPM family. We recently demonstrated that PPM1H can dephosphorylate Smad1 in the cytoplasm and block BMP signaling responses in cellular assays. Here we describe in vitro method showing that PPM1H is a bona fide phosphatase for Smad1/5/8. PPM1H is produced as GST fusion protein in E. coli, and purified against glutathione sepharose beads. Bacterially purified recombinant PPM1H possesses phosphatase activity toward artificial substrate para-nitrophenyl phosphate (pNPP). Recombinant PPM1H also dephosphorylates immuno-purified phosphorylated Smad1 in test tubes. These direct in vitro phosphatase assays provide convincing evidence demonstrating the role of PPM1H as a specific phosphatase for P-Smad1.
Recent Data Analysis of Carbon ACtivation
NASA Astrophysics Data System (ADS)
Jiang, Hui Ming; Smith, Elizabeth; Padalino, Stephen; Baumgart, Leigh; Suny Geneseooltz, Katie; Colburn, Robyn; Fuschino, Julia
A method for measuring tertiary neutrons produced in Inertial Confinement Fusion reactions has been developed using carbon activation. Ultra pure samples of carbon, free from positron-emitting contaminants must be used in the detection. Our primary goal has been to reduce the contamination level by refining purification and packaging procedures. This process involves baking the disks in a vacuum oven to 1000¢XC @ 200 microns for a prescribed bake time without exposing the disks to nitrogen in the air which is a major contaminant. Recent experiments were conducted to determine the optimal bake time for purification. Disks were baked for varying times, from one hour to five hours, and then exposed to high-neutron-yield ( 5 x 1013) shots on OMEGA. Data collected was normalized to the same time interval and the same primary neutron yield, and no significant difference in the number of background counts was seen. Experimental results also indicated that disks that were exposed to air for short time intervals showed a significant increase in the number of contamination counts. This further supports our findings that the gaseous diffusion through graphite disks is very high. Experimental results of these findings will be presented. Research funded in part by the United States Department of Energy.
Analysis of supercooling activities of surfactants.
Kuwabara, Chikako; Terauchi, Ryuji; Tochigi, Hiroshi; Takaoka, Hisao; Arakawa, Keita; Fujikawa, Seizo
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).
Stellar Activity from Analysis of Planetary Transits
NASA Astrophysics Data System (ADS)
Valio, A.
About a third of the extra solar planets discovered so far transit their host star. During the eclipse of the star by its orbiting planet, spots on the surface of the star may be occulted, causing small variation in the transit lightcurve. These variations can be modelled using a method developed by Silva (2003) that yields the starspots physical properties such as size, position, temperature (or intensity), and lifetime. Just like Galileo did four centuries ago for the Sun, from the spot analysis it is also possible to calculate the stellar rotation period and whether it presents or not differential rotation. The mean rotation period of the star is obtained from the out-of-transit light curve modulation, whereas the value of the rotation period at the latitude of the transit is determined from the successive transits of the same spot. Adopting a solar-like rotation profile as a function of latitude, the differential rotation for several CoRoT planet hosting stars is estimated.
Escape from neutralization by the respiratory syncytial virus-specific neutralizing monoclonal antibody palivizumab is driven by changes in on-rate of binding to the fusion protein.
Bates, John T; Keefer, Christopher J; Slaughter, James C; Kulp, Daniel W; Schief, William R; Crowe, James E
The role of binding kinetics in determining neutralizing potency for antiviral antibodies is poorly understood. While it is believed that increased steady-state affinity correlates positively with increased virus-neutralizing activity, the relationship between association or dissociation rate and neutralization potency is unclear. We investigated the effect of naturally-occurring antibody resistance mutations in the RSV F protein on the kinetics of binding to palivizumab. Escape from palivizumab-mediated neutralization of RSV occurred with reduced association rate (Kon) for binding to RSV F protein, while alteration of dissociation rate (Koff) did not significantly affect neutralizing activity. Interestingly, linkage of reduced Kon with reduced potency mirrored the effect of increased Kon found in a high-affinity enhanced potency palivizumab variant (motavizumab). These data suggest that association rate is the dominant factor driving neutralization potency for antibodies to RSV F protein antigenic site A and determines the potency of antibody somatic variants or efficiency of escape of viral glycoprotein variants.
Applying an Activity System to Online Collaborative Group Work Analysis
ERIC Educational Resources Information Center
Choi, Hyungshin; Kang, Myunghee
This study determines whether an activity system provides a systematic framework to analyse collaborative group work. Using an activity system as a unit of analysis, the research examined learner behaviours, conflicting factors and facilitating factors while students engaged in collaborative work via asynchronous computer-mediated communication.…
Spatiotemporal Data Mining, Analysis, and Visualization of Human Activity Data
ERIC Educational Resources Information Center
Li, Xun
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…
Project-Based Language Learning: An Activity Theory Analysis
ERIC Educational Resources Information Center
Gibbes, Marina; Carson, Lorna
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…
Understanding Tensions: Activity Systems Analysis of Cross-Continental Collaboration
ERIC Educational Resources Information Center
Ryder, LanHui Zhang; Yamagata-Lynch, Lisa
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 University (SDNU),…
Overview af MSFC's Applied Fluid Dynamics Analysis Group Activities
NASA Technical Reports Server (NTRS)
Garcia, Roberto; Griffin, Lisa; Williams, Robert
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.
PIXE and neutron activation methods in human hair material analysis
NASA Astrophysics Data System (ADS)
Bǎdicǎ, T.; Ciortea, C.; Cojocaru, V.; Ivaşcu, M.; Petrovici, A.; Popa, A.; Popescu, I.; Sǎlǎgean, M.; Spiridon, S.
In order to compare some of the nuclear methods in human hair material analysis, proton induced X-ray excitation and variant techniques of neutron activation analysis have been used. The elemental concentrations are compared with the IAEA-Vienna certified values. The efficiency and reliability of the methods used are briefly discussed.
Molecular clips and tweezers hosting neutral guests.
Hardouin-Lerouge, Marie; Hudhomme, Piétrick; Sallé, Marc
Intense current interest in supramolecular chemistry is devoted to the construction of molecular assemblies displaying controlled molecular motion associated to recognition. On this ground, molecular clips and tweezers have focused an increasing attention. This tutorial review points out the recent advances in the construction of always more sophisticated molecular clips and tweezers, illustrating their remarkably broad structural variety and focusing on their binding ability towards neutral guests. A particular attention is brought to recent findings in dynamic molecular tweezers whose recognition ability can be regulated by external stimuli. Porphyrin-based systems will not be covered here as this very active field has been recently reviewed.
Photodetachment process for beam neutralization
Fink, Joel H. [Livermore, CA; Frank, Alan M. [Livermore, CA
A process for neutralization of accelerated ions employing photo-induced charge detachment. The process involves directing a laser beam across the path of a negative ion beam such as to effect photodetachment of electrons from the beam ions. The frequency of the laser beam employed is selected to provide the maximum cross-section for the photodetachment process.
Photodetachment process for beam neutralization
Fink, J.H.; Frank, A.M.
A process for neutralization of accelerated ions employing photo-induced charge detachment is disclosed. The process involves directing a laser beam across the path of a negative ion beam such as to effect photodetachment of electrons from the beam ions. The frequency of the laser beam employed is selected to provide the maximum cross-section for the photodetachment process. 2 figs.
Low energy neutral atom imaging
McComas, D.J.; Funsten, H.O.; Gosling, J.T.; Moore, K.R.; Thomsen, M.F.
Energetic neutral atom (ENA) and low energy neutral atom (LENA) imaging of space plasmas are emerging new technology which promises to revolutionize the way we view and understand large scale space plasma phenomena and dynamics. ENAs and LENAs are produced in the magnetosphere by charge exchange between energetic and plasma ions and cold geocoronal neutrals. While imaging techniques have been previously developed for observing ENAs, with energies above several tens of keV, most of the ions found in the terrestrial magnetosphere have lower energies. We recently suggested that LENAs could be imaged by first converting the neutrals to ions and then electrostatically analyzing them to reject the UV background. In this paper we extend this work to examine in detail the sensor elements needed to make an LENA imager. These elements are (1) a biased collimator to remove the ambient plasma ions and electrons and set the azimuthal field-of-view; (2) a charge modifier to convert a portion of the incident LENAs to ions; (3) an electrostatic analyzer to reject UV light and set the energy passband; and (4) a coincidence detector to measure converted LENAs while rejecting noise and penetrating radiation. We also examine the issue of LENA imager sensitivity and describe ways of optimizing sensitivity in the various sensor components. Finally, we demonstrate in detail how these general considerations are implemented by describing one relatively straightforward design based on a hemispherical electrostatic analyzer.
Energy distributions of sputtered copper neutrals and ions
NASA Technical Reports Server (NTRS)
Lundquist, T. R.
Direct quantitative analysis of surfaces by secondary ion mass spectrometry will depend on an understanding of the yield ratio of ions to neutrals. This ratio as a function of the energy of the sputtered particles has been obtained for a clean polycrystalline copper surface sputtered by 1000-3000 eV Ar(+). The energy distributions of both neutral and ionized copper were measured with a retarding potential analyzer using potential modulation differentiation and signal averaging. The maximum for both distributions is identical and occurs near 2.5 eV. The energy distributions of neutrals is more sharply peaked than that of the ions, presumably as a consequence of more efficient nutralization of slow escaping ions by the mobile electrons of copper. The ion-neutral ratio is compared with results from various ionization models.
Neutralization of Home Made Explosives (HME) Hazards
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Neutral atomic carbon in dense molecular clouds
NASA Technical Reports Server (NTRS)
Zmuidzinas, J.; Betz, A. L.; Boreiko, R. T.; Goldhaber, D. M.
The 370 micron 3P2-3P1 fine-structure line of neutral carbon was detected in seven sources: OMC 1, NGC 2024, S140, W3, DR 21, M17, and W51. Simultaneous analysis of J = 2-1 data and available observations of the J = 1-0 line make it possible to deduce optical depths and excitation temperatures for these lines. These data indicate that both C I lines are likely to be optically thin, and that the ratio of C I to CO column densities in these clouds is typically about 0.1.
Surface Electromyographic Analysis of Core Trunk and Hip Muscles During Selected Rehabilitation Exercises in the Side-Bridge to Neutral Spine Position
Youdas, James W.; Boor, Mallory M.P.; Darfler, Arynn L.; Koenig, Margaret K.; Mills, Katherine M.; Hollman, John H.
Background: Strengthening of core hip, trunk, and abdominal muscles has been utilized with injury prevention and low back pain and has the potential to improve athletic performance. Hypothesis: During a side-bridge, trunk and thigh muscles on the ipsilateral weightbearing side would produce greater activation than their counterparts on the contralateral nonweightbearing side. Study Design: Descriptive laboratory study. Methods: Twelve females and 13 males participated. Electromyography (EMG) signals were gathered for 5 right-sided muscles (rectus abdominis [RA], external oblique [EO], longissimus thoracis [LT], lumbar multifidus [LM], and gluteus medius [GM]) during 3 repetitions of 4 side-bridging exercises (trunk-elevated side support [TESS], foot-elevated side support [FESS], clamshell, and rotational side-bridge [RSB]) performed bilaterally in random order using surface electrodes. EMG signals were normalized to peak activity in maximum voluntary isometric contraction (MVIC) trials and expressed as a percentage. Descriptive EMG data were calculated for EMG recruitment (% MVIC) and compared between right side up and right side down conditions and between exercises with 2-way repeated-measures analyses of variance at α = 0.05. Results: RSB created the most muscle activation in 3 of 4 recorded trunk muscles (RA, 43.9% MVIC; EO, 62.8 % MVIC; and LT, 41.3% MVIC). Activation of the GM exceeded 69% MVIC for TESS, FESS, and RSB. With the exception of the RA in RSB and LT in TESS, recruitment within muscles of the ipsilateral weightbearing trunk and thigh (% MVIC) was significantly greater than their counterparts on the nonweightbearing trunk and thigh for all muscles during the side-bridge exercise conditions. Conclusion: Muscle recruitment was greater within muscles of the ipsilateral weightbearing trunk and thigh for all examined muscles except RA during RSB and LT during TESS. Activation at or above 50% MVIC is needed for strengthening. Activation of the GM and EO
Low frequency of broadly neutralizing HIV antibodies during chronic infection even in quaternary epitope targeting antibodies containing large numbers of somatic mutations.
Hicar, Mark D; Chen, Xuemin; Kalams, Spyros A; Sojar, Hakimuddin; Landucci, Gary; Forthal, Donald N; Spearman, Paul; Crowe, James E
Neutralizing antibodies (Abs) are thought to be a critical component of an appropriate HIV vaccine response. It has been proposed that Abs recognizing conformationally dependent quaternary epitopes on the HIV envelope (Env) trimer may be necessary to neutralize diverse HIV strains. A number of recently described broadly neutralizing monoclonal Abs (mAbs) recognize complex and quaternary epitopes. Generally, many such Abs exhibit extensive numbers of somatic mutations and unique structural characteristics. We sought to characterize the native antibody (Ab) response against circulating HIV focusing on such conformational responses, without a prior selection based on neutralization. Using a capture system based on VLPs incorporating cleaved envelope protein, we identified a selection of B cells that produce quaternary epitope targeting Abs (QtAbs). Similar to a number of broadly neutralizing Abs, the Ab genes encoding these QtAbs showed extensive numbers of somatic mutations. However, when expressed as recombinant molecules, these Abs failed to neutralize virus or mediate ADCVI activity. Molecular analysis showed unusually high numbers of mutations in the Ab heavy chain framework 3 region of the variable genes. The analysis suggests that large numbers of somatic mutations occur in Ab genes encoding HIV Abs in chronically infected individuals in a non-directed, stochastic, manner.
Low frequency of broadly neutralizing HIV antibodies during chronic infection even in quaternary epitope targeting antibodies containing large numbers of somatic mutations
Hicar, Mark D.; Chen, Xuemin; Kalams, Spyros A.; Sojar, Hakimuddin; Landucci, Gary; Forthal, Donald N.; Spearman, Paul; Crowe, James E.
Neutralizing antibodies (Abs) are thought to be a critical component of an appropriate HIV vaccine response. It has been proposed that Abs recognizing conformationally dependent quaternary epitopes on the HIV envelope (Env) trimer may be necessary to neutralize diverse HIV strains. A number of recently described broadly neutralizing monoclonal Abs (mAbs) recognize complex and quaternary epitopes. Generally, many such Abs exhibit extensive numbers of somatic mutations and unique structural characteristics. We sought to characterize the native antibody (Ab) response against circulating HIV focusing on such conformational responses, without a prior selection based on neutralization. Using a capture system based on VLPs incorporating cleaved envelope protein, we identified a selection of B cells that produce quaternary epitope targeting Abs (QtAbs). Similar to a number of broadly neutralizing Abs, the Ab genes encoding these QtAbs showed extensive numbers of somatic mutations. However, when expressed as recombinant molecules, these Abs failed to neutralize virus or mediate ADCVI activity. Molecular analysis showed unusually high numbers of mutations in the Ab heavy chain framework 3 region of the variable genes. The analysis suggests that large numbers of somatic mutations occur in Ab genes encoding HIV Abs in chronically infected individuals in a non-directed, stochastic, manner. PMID:26748387
Lack of protection following passive transfer of polyclonal highly functional low-dose non-neutralizing antibodies.
Dugast, Anne-Sophie; Chan, Ying; Hoffner, Michelle; Licht, Anna; Nkolola, Joseph; Li, Hualin; Streeck, Hendrik; Suscovich, Todd J; Ghebremichael, Musie; Ackerman, Margaret E; Barouch, Dan H; Alter, Galit
Recent immune correlates analysis from the RV144 vaccine trial has renewed interest in the role of non-neutralizing antibodies in mediating protection from infection. While neutralizing antibodies have proven difficult to induce through vaccination, extra-neutralizing antibodies, such as those that mediate antibody-dependent cellular cytotoxicity (ADCC), are associated with long-term control of infection. However, while several non-neutralizing monoclonal antibodies have been tested for their protective efficacy in vivo, no studies to date have tested the protective activity of naturally produced polyclonal antibodies from individuals harboring potent ADCC activity. Because ADCC-inducing antibodies are highly enriched in elite controllers (EC), we passively transferred highly functional non-neutralizing polyclonal antibodies, purified from an EC, to assess the potential impact of polyclonal non-neutralizing antibodies on a stringent SHIV-SF162P3 challenge in rhesus monkeys. Passive transfer of a low-dose of ADCC inducing antibodies did not protect from infection following SHIV-SF162P3 challenge. Passively administered antibody titers and gp120-specific, but not gp41-specific, ADCC and antibody induced phagocytosis (ADCP) were detected in the majority of the monkeys, but did not correlate with post infection viral control. Thus these data raise the possibility that gp120-specific ADCC activity alone may not be sufficient to control viremia post infection but that other specificities or Fc-effector profiles, alone or in combination, may have an impact on viral control and should be tested in future passive transfer experiments.
Neutron activation analysis for antimetabolites. [in food samples
NASA Technical Reports Server (NTRS)
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.
Recent Progress toward Engineering HIV-1-Specific Neutralizing Monoclonal Antibodies
Sun, Ming; Li, Yue; Zheng, Huiwen; Shao, Yiming
The recent discoveries of broadly potent neutralizing human monoclonal antibodies represent a new generation of antiretrovirals for the treatment and prophylaxis. Antibodies are generally considered more effective and safer and have been proved to provide passive protection against mucosal challenge in humanized mice and macaques. Several neutralizing Abs could protect animals against HIV-1 but are not effective when used in an established infected model for therapy. In order to overcome the limitation of antiviral activities, multiple antibody-engineering technologies have been explored to generate "the better" neutralizing antibodies against HIV-1 since bNAbs attack viral entry by various mechanisms. Thus, a promising direction of research is to discover and exploit rational antibody combination or engineered antibodies (eAbs) as potential candidate therapeutics against HIV-1. It has been reported that inclusion of fusion-neutralizing antibodies in a set of bNAbs could improve their overall activities and neutralizing spectrum. Here, we review several routes for engineering bNAbs, such as design and generation of bispecific antibodies, specific glycosylation of antibodies to enhance antiviral activity, and variable region-specific modification guided by structure and computer, as well as reviewing antibody-delivery technologies by non-viral vector, viral vector, and human hematopoietic stem/progenitor cells transduced with a lentiviral construct. We also discuss the optimized antiviral activities and benefits of these strategy and potential mechanisms. PMID:27746780
Characterization of the wild type and truncated form of a neutral GH10 xylanase from Coprinus cinereus: Roles of C-terminal basic amino acid-rich extension in its SDS resistance, thermostability and activity.
Hu, Hang; Chen, Kaixiang; Li, Lulu; Long, Liangkun; Ding, Shaojun
A neutral xylanase (Ccxyn) was identified from Coprinus cinereus. It has a single GH10 catalytic domain with a basic amino acid-rich extension (PVRRK) at C-terminus. In this study, the wild-type (Ccxyn) and C-terminus truncated xylanase (CcXynΔ5C) were heterologously expressed in Pichia pastoris and their characteristics were comparatively analyzed with aims to examine the effect of this extension on the enzyme function. The circular dichorism (CD) analysis indicated that both enzymes in general had a similar structure, but CcXynΔ5C contained less α-helices (42.9%) and more random coil contents (35.5%) than CcXyn (47.0% and 32.8%, respectively). Both enzymes had same pH (7.0) and temperature (45°C) optima, and similar substrate specificity on different xylans. They all hydrolyzed beechwood xylan primarily to xylobiose and xylotriose. The amounts of xylobiose and xylotriose account for 91.5% and 92.2 % (w/w) of total xylooligosaccharides (XOS) generated from beechwood by Ccxyn and CcxynΔ5C, respectively. However, truncation of C-terminal 5-amino acids extension significantly improves its thermostability, SDS resistance and pH stability at pH 6.0-9.0. Furthermore, CcxynΔ5C exhibited much lower Km value than Ccxyn (0.27 mg ml⁻¹ vs 0.83 mg ml⁻¹), therefore, the catalytic efficiency of CcXynΔ5C was 2.4 times higher than that of CcXyn. These properties make CcxynΔ5C a good model for the structure-function study of (α/β)₈-barrel folded enzyme and a promising candidate for various applications, especially in the detergent industry and XOS production.
Thermostability of Well-Ordered HIV Spikes Correlates with the Elicitation of Autologous Tier 2 Neutralizing Antibodies.
Feng, Yu; Tran, Karen; Bale, Shridhar; Kumar, Shailendra; Guenaga, Javier; Wilson, Richard; de Val, Natalia; Arendt, Heather; DeStefano, Joanne; Ward, Andrew B; Wyatt, Richard T
In the context of HIV vaccine design and development, HIV-1 spike mimetics displaying a range of stabilities were evaluated to determine whether more stable, well-ordered trimers would more efficiently elicit neutralizing antibodies. To begin, in vitro analysis of trimers derived from the cysteine-stabilized SOSIP platform or the uncleaved, covalently linked NFL platform were evaluated. These native-like trimers, derived from HIV subtypes A, B, and C, displayed a range of thermostabilities, and were "stress-tested" at varying temperatures as a prelude to in vivo immunogenicity. Analysis was performed both in the absence and in the presence of two different adjuvants. Since partial trimer degradation was detected at 37°C before or after formulation with adjuvant, we sought to remedy such an undesirable outcome. Cross-linking (fixing) of the well-ordered trimers with glutaraldehyde increased overall thermostability, maintenance of well-ordered trimer integrity without or with adjuvant, and increased resistance to solid phase-associated trimer unfolding. Immunization of unfixed and fixed well-ordered trimers into animals revealed that the elicited tier 2 autologous neutralizing activity correlated with overall trimer thermostability, or melting temperature (Tm). Glutaraldehyde fixation also led to higher tier 2 autologous neutralization titers. These results link retention of trimer quaternary packing with elicitation of tier 2 autologous neutralizing activity, providing important insights for HIV-1 vaccine design.
The neutralization of interferons by antibody. I. Quantitative and theoretical analyses of the neutralization reaction in different bioassay systems.
Grossberg, S E; Kawade, Y; Kohase, M; Yokoyama, H; Finter, N
The highly specific ability of antibodies to inhibit the biologic activity of cytokines or other therapeutic proteins is widely used in research and a subject of increasing clinical importance. The need exists for a standardized approach to the reporting of neutralizing antibody potency soundly based on theoretical and practical considerations and tested by experimental data. Pursuant to the original studies of Kawade on the theoretical and functional aspects of neutralization of interferons (IFN), experimental data were obtained by different laboratories employing varied methodology to address two hypotheses concerning the nature of IFN neutralization reactions, based on a derived formula that allows expression of neutralizing power as the reduction of 10 laboratory units (LU)/ml to 1 LU/ml, the end point of most bioassays. Two hypotheses are posed: (1) antibody acts to neutralize a fixed amount of biologically active IFN molecules, or (2) antibody reduces IFN activity in a set ratio of added/residual biologically active IFN. The first, or fixed amount, hypothesis relates to the reactivity of high-affinity antibodies neutralizing equimolar amounts of antigen, whereas the second, or constant proportion, hypothesis postulates a reduction in the ratio of total added IFN to residual active IFN molecules, such as a low-affinity antibody might exhibit. Analyses of data of the neutralization of IFN-alpha and IFN-beta are presented, employing human polyclonal antibodies and murine monoclonal antibodies (mAb). The theoretical constructs of Kawade are extended in the Appendix and correlated with new experimental data in the text. The data clearly indicate that the low-antibody affinity, constant proportion hypothesis, rather than the high-antibody affinity, fixed amount hypothesis, is applicable, if the bioassay is sensitive to IFN. The findings presented here and in the following paper (pp. 743-755, this issue) taken together provide the basis for a standardized method of
Neutralization of Epstein-Barr Virus by Nonimmune Human Serum
Nemerow, Glen R.; Jensen, Fred C.; Cooper, Neil R.
These studies were carried out to investigate the mechanism of neutralization of purified Epstein-Barr virus (EBV) by fresh human serum from normal individuals lacking antibody to the EBV viral capsid (VCA) and nuclear antigens (EBNA). Such individuals thus lack serological evidence of immunity to EBV. Although an enzyme-linked immunosorbent assay (ELISA) with highly purified immobilized EBV detected low levels of IgG antibody reactive with EBV in these normal nonimmune sera, this antibody failed to neutralize EBV in the absence of complement. Studies with depleted sera and mixtures of purified complement proteins at physiologic concentrations showed that the IgG antibody and C1, C4, C2, and C3 of the classical pathway were able to fully neutralize EBV. Mixtures of the purified components of the alternative pathway at physiologic concentrations failed to neutralize purified EBV in the presence or absence of the antibody and the alternative pathway did not potentiate classical pathway-mediated neutralization. No evidence for a requirement for C8 was obtained, precluding lysis as the mechanism of neutralization. Since C3 deposition on the viral surface accompanied classical pathway activation, viral neutralization is most likely secondary to the accumulation of complement protein on the viral surface. A coating of protein on the virus could interfere with attachment to, or penetration of potentially susceptible cells. Experiments were undertaken to determine the specificity of the IgG antibody in the sera of EBV nonimmune individuals which, together with complement, neutralized EBV. Both purified EBV and herpes simplex I (HSV-1) absorbed the EBV ELISA reactivity and EBV-neutralizing activity of nonimmune sera, whereas another member of the herpesvirus group, cytomegalovirus, was inactive in this regard. HSV-1 was quantitatively more efficient than EBV in absorbing reactivity, a finding that indicates that the antibody has a higher affinity for HSV-1 than for EBV
Crystallization and preliminary analysis of active nitroalkane oxidase in three crystal forms
Nagpal, Akanksha; Valley, Michael P.; Fitzpatrick, Paul F.; Orville, Allen M.
Nitroalkane oxidase (NAO), a flavoprotein cloned and purified from Fusarium oxysporum, catalyzes the oxidation of neutral nitroalkanes to the corresponding aldehydes or ketones, with the production of H2O2 and nitrite. In this paper, the crystallization and preliminary X-ray data analysis of three crystal forms of active nitroalkane oxidase are described. The first crystal form belongs to a trigonal space group (either P3121 or P3221, with unit-cell parameters a = b = 103.8, c = 487.0 Å) and diffracts to at least 1.6 Å resolution. Several data sets were collected using 2θ and κ geometry in order to obtain a complete data set to 2.07 Å resolution. Solvent-content and Matthews coefficient analysis suggests that crystal form 1 contains two homotetramers per asymmetric unit. Crystal form 2 (P212121; a = 147.3, b = 153.5, c = 169.5 Å) and crystal form 3 (P31 or P32; a = b = 108.9, c = 342.5 Å) are obtained from slightly different conditions and also contain two homotetramers per asymmetric unit, but have different solvent contents. A three-wavelength MAD data set was collected from selenomethionine-enriched NAO (SeMet-NAO) in crystal form 3 and will be used for phasing. PMID:15272176
A statistical analysis of eruptive activity on Mount Etna, Sicily
NASA Astrophysics Data System (ADS)
Smethurst, Lucy; James, Mike R.; Pinkerton, Harry; Tawn, Jonathan A.
A rigorous analysis of the timing and location of flank eruptions of Mount Etna on Sicily is important for the creation of hazard maps of the densely populated area surrounding the volcano. In this paper, we analyse the temporal, volumetric and spatial data on eruptive activity on Etna. Our analyses are based on the two most recent and robust historical data catalogues of flank eruption activity on Etna, with one from 1669 to 2008 and the other from 1610 to 2008. We use standard statistical methodology and modelling techniques, though a number of features are new to the analysis of eruption data. Our temporal analysis reveals that flank eruptions on Mount Etna between 1610 and 2008 follow an inhomogeneous Poisson process, with intensity of eruptions increasing nearly linearly since the mid-1900s. Our temporal analysis reveals no evidence of cyclicity over this period. An analysis of volumetric lava flow rates shows a marked increase in activity since 1971. This increase, which coincides with the formation of the Southeast Crater (SEC), appears to be related to increased activity on and around the SEC. This has significant implications for hazard analysis on Etna.
Isolation and immunizations with hepatitis A viral structural proteins: induction of antiprotein, antiviral, and neutralizing responses.
Hughes, J V; Stanton, L W
An immune affinity purification procedure for hepatitis A virus (HAV) was designed which yielded milligram quantities of the virus with greater than 95% purity. The major structural proteins VP-1, VP-2, and VP-3 were isolated from the purified virus by electroelution from sodium dodecyl sulfate-polyacrylamide gels and used to immunize Lewis rats (three to four doses, 10 to 15 micrograms per dose). The two Lewis rats immunized with VP-1 developed a strong antibody response to VP-1, as determined by Western blot analysis and immune precipitation of the denatured protein. These animals also developed a good antibody response to the whole virus, as demonstrated by a positive response in a competitive radioimmunoassay (HAV antibody test) and by precipitation of the whole virus. In addition, both animals developed a low titer neutralizing antibody to HAV, as demonstrated by an in vitro cell culture assay. While the two rats receiving VP-2 developed only minimal responses to the protein and to the virus by the same assays described above, one of the two developed a significant neutralizing antibody to HAV. The immunization of one Lewis rat with VP-3 induced a good antibody response to both denatured protein and the whole virus. This serum sample was also demonstrated to neutralize the viral infectivity. Finally, two rabbits that had received inoculations of sodium dodecyl sulfate and heat-disrupted HAV (containing 20 to 30 micrograms of each protein per dose) developed good antiprotein responses to all of the proteins and good antiviral responses, including a consistently significant neutralizing activity. The neutralizing antibody responses suggest that the structural proteins of HAV, or a portion of them, could provide the basis for a subunit vaccine for HAV.
Preferential Amygdala Reactivity to the Negative Assessment of Neutral Faces
Blasi, Giuseppe; Hariri, Ahmad R.; Alce, Guilna; Taurisano, Paolo; Sambataro, Fabio; Das, Saumitra; Bertolino, Alessandro; Weinberger, Daniel R.; Mattay, Venkata S.
Background Prior studies suggest that the amygdala shapes complex behavioral responses to socially ambiguous cues. We explored human amygdala function during explicit behavioral decision making about discrete emotional facial expressions that can represent socially unambiguous and ambiguous cues. Methods During functional magnetic resonance imaging, 43 healthy adults were required to make complex social decisions (i.e., approach or avoid) about either relatively unambiguous (i.e., angry, fearful, happy) or ambiguous (i.e., neutral) facial expressions. Amygdala activation during this task was compared with that elicited by simple, perceptual decisions (sex discrimination) about the identical facial stimuli. Results Angry and fearful expressions were more frequently judged as avoidable and happy expressions most often as approachable. Neutral expressions were equally judged as avoidable and approachable. Reaction times to neutral expressions were longer than those to angry, fearful, and happy expressions during social judgment only. Imaging data on stimuli judged to be avoided revealed a significant task by emotion interaction in the amygdala. Here, only neutral facial expressions elicited greater activity during social judgment than during sex discrimination. Furthermore, during social judgment only, neutral faces judged to be avoided were associated with greater amygdala activity relative to neutral faces that were judged as approachable. Moreover, functional coupling between the amygdala and both dorsolateral prefrontal (social ju