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Sample records for oligomerized pool engineering

  1. Rapid mutation of endogenous zebrafish genes using zinc finger nucleases made by Oligomerized Pool ENgineering (OPEN.

    Directory of Open Access Journals (Sweden)

    Jonathan E Foley

    Full Text Available BACKGROUND: Customized zinc finger nucleases (ZFNs form the basis of a broadly applicable tool for highly efficient genome modification. ZFNs are artificial restriction endonucleases consisting of a non-specific nuclease domain fused to a zinc finger array which can be engineered to recognize specific DNA sequences of interest. Recent proof-of-principle experiments have shown that targeted knockout mutations can be efficiently generated in endogenous zebrafish genes via non-homologous end-joining-mediated repair of ZFN-induced DNA double-stranded breaks. The Zinc Finger Consortium, a group of academic laboratories committed to the development of engineered zinc finger technology, recently described the first rapid, highly effective, and publicly available method for engineering zinc finger arrays. The Consortium has previously used this new method (known as OPEN for Oligomerized Pool ENgineering to generate high quality ZFN pairs that function in human and plant cells. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that OPEN can also be used to generate ZFNs that function efficiently in zebrafish. Using OPEN, we successfully engineered ZFN pairs for five endogenous zebrafish genes: tfr2, dopamine transporter, telomerase, hif1aa, and gridlock. Each of these ZFN pairs induces targeted insertions and deletions with high efficiency at its endogenous gene target in somatic zebrafish cells. In addition, these mutations are transmitted through the germline with sufficiently high frequency such that only a small number of fish need to be screened to identify founders. Finally, in silico analysis demonstrates that one or more potential OPEN ZFN sites can be found within the first three coding exons of more than 25,000 different endogenous zebrafish gene transcripts. CONCLUSIONS AND SIGNIFICANCE: In summary, our study nearly triples the total number of endogenous zebrafish genes successfully modified using ZFNs (from three to eight and suggests that OPEN

  2. Predicting success of oligomerized pool engineering (OPEN for zinc finger target site sequences

    Directory of Open Access Journals (Sweden)

    Goodwin Mathew J

    2010-11-01

    Full Text Available Abstract Background Precise and efficient methods for gene targeting are critical for detailed functional analysis of genomes and regulatory networks and for potentially improving the efficacy and safety of gene therapies. Oligomerized Pool ENgineering (OPEN is a recently developed method for engineering C2H2 zinc finger proteins (ZFPs designed to bind specific DNA sequences with high affinity and specificity in vivo. Because generation of ZFPs using OPEN requires considerable effort, a computational method for identifying the sites in any given gene that are most likely to be successfully targeted by this method is desirable. Results Analysis of the base composition of experimentally validated ZFP target sites identified important constraints on the DNA sequence space that can be effectively targeted using OPEN. Using alternate encodings to represent ZFP target sites, we implemented Naïve Bayes and Support Vector Machine classifiers capable of distinguishing "active" targets, i.e., ZFP binding sites that can be targeted with a high rate of success, from those that are "inactive" or poor targets for ZFPs generated using current OPEN technologies. When evaluated using leave-one-out cross-validation on a dataset of 135 experimentally validated ZFP target sites, the best Naïve Bayes classifier, designated ZiFOpT, achieved overall accuracy of 87% and specificity+ of 90%, with an ROC AUC of 0.89. When challenged with a completely independent test set of 140 newly validated ZFP target sites, ZiFOpT performance was comparable in terms of overall accuracy (88% and specificity+ (92%, but with reduced ROC AUC (0.77. Users can rank potentially active ZFP target sites using a confidence score derived from the posterior probability returned by ZiFOpT. Conclusion ZiFOpT, a machine learning classifier trained to identify DNA sequences amenable for targeting by OPEN-generated zinc finger arrays, can guide users to target sites that are most likely to function

  3. Gelatin-based biomaterial engineering with anhydride-containing oligomeric cross-linkers.

    Science.gov (United States)

    Loth, Tina; Hötzel, Rudi; Kascholke, Christian; Anderegg, Ulf; Schulz-Siegmund, Michaela; Hacker, Michael C

    2014-06-01

    Chemically cross-linked gelatin hydrogels are versatile cell-adhesive hydrogel materials that have been established for a variety of biomedical applications. The most prominent cross-linker is glutaraldehyde, which, however, has been described to cause compatibility problems and loss of microscopic but relevant structural features. A recently developed oligomeric cross-linker that contains anhydride functionalities was evaluated as cross-linker for the fabrication of gelatin-based hydrogels and microparticles. In a fast curing reaction, hydrogels composed of gelatin and oligomeric cross-linker were fabricated with good conversion over a wide concentration range of constituents and with cross-linkers of different anhydride contents. Hydrogel properties, such as dry weight and mechanics, could be controlled by hydrogel composition and rheological properties correlated to elastic moduli from 1 to 10 kPa. The gels were shown to be cytocompatible and promoted cell adhesion. In soft formulations, cells migrated into the hydrogel bulk. Gelatin microparticles prepared by a standard water-in-oil emulsion technique were also treated with the novel oligomers, and cross-linking degrees matching those obtained with glutaraldehyde were obtained. At the same time, fewer interparticular cross-links were observed. Fluorescein-derivatized cross-linkers yielded labeled microparticles in a concentration-dependent manner. The oligomeric cross-linkers are presented as an efficient and possibly more functional and compatible alternative to glutaraldehyde. The engineered hydrogel materials hold potential for various biomedical applications.

  4. ITC-Euromaster Course Pool for AEC Engineers

    Directory of Open Access Journals (Sweden)

    Andrej Tibaut

    2013-05-01

    Full Text Available The paper describes the ITC-Euromaster Course Pool. It is a concept that powers e-learning environment for conducting an international Master’s level programme in information technologies for Architecture, Engineering and Construction (AEC. The e-learning environment integrates resources (units of study, learning management system, virtual classroom, teachers and students from five European universities. In the paper following aspects of the system are presented: concept and methodology of the Course Pool, technology for e-learning and evaluation results.

  5. Simulation modeling of pooling for combinatorial protein engineering.

    Science.gov (United States)

    Polizzi, Karen M; Spencer, Cody U; Dubey, Anshul; Matsumura, Ichiro; Lee, Jay H; Realff, Matthew J; Bommarius, Andreas S

    2005-12-01

    Pooling in directed-evolution experiments will greatly increase the throughput of screening systems, but important parameters such as the number of good mutants created and the activity level increase of the good mutants will depend highly on the protein being engineered. The authors developed and validated a Monte Carlo simulation model of pooling that allows the testing of various scenarios in silico before starting experimentation. Using a simplified test system of 2 enzymes, betagalactosidase (supermutant, or greatly improved enzyme) and beta-glucuronidase (dud, or enzyme with ancestral level of activity), the model accurately predicted the number of supermutants detected in experiments within a factor of 2. Additional simulations using more complex activity distributions show the versatility of the model. Pooling is most suited to cases such as the directed evolution of new function in a protein, where the background level of activity is minimized, making it easier to detect small increases in activity level. Pooling is most successful when a sensitive assay is employed. Using the model will increase the throughput of screening procedures for directed-evolution experiments and thus lead to speedier engineering of proteins.

  6. Zn(2+) site engineering at the oligomeric interface of the dopamine transporter

    DEFF Research Database (Denmark)

    Norgaard-Nielsen, Kristine; Norregaard, Lene; Hastrup, Hanne

    2002-01-01

    of the dopamine transporter (DAT) corresponding to the external end of transmembrane segment 6. Upon binding to this site, which involves a histidine inserted in position 310 (V310H) and the endogenous Cys306 within the same DAT molecule, Zn(2+) potently inhibits [(3)H]dopamine uptake. These data provide indirect...... evidence that conformational changes critical for the translocation process may occur at the interface between two transporter molecules in the oligomeric structure....

  7. Automated management of engineering infrastructure of pools of different purpose

    Directory of Open Access Journals (Sweden)

    Shirokov Lev Alekseevich

    2016-10-01

    Full Text Available Pools play an important role in people’s life. They answer people’s demand in rest and improve their health. At the same time pools are rather important for industrial use, for example in construction industry. In order to solve different construction problems it is essential to investigate the influence of microclimatic parameters on construction materials and structures. For this aim pools are in demand as special test sites for construction materials and structures in different environmental conditions including the case of a direct water impact. The efficient use of pools presupposes the necessity of constant hydroclimatic contro: air humidity and temperature, water temperature, chemical composition of water and air. Classification of pools of different purposes is presented in the article. The author considers the main problems of operation of pools as objects with complicated air-and-water environment. The questions of maintaining optimal microclimatic parameters in a pool are considered. The necessity of use of the control system of a microclimate, its efficiency, profitability and social effect of its implementation is described. A mathematical model of the thermal mode of a pool area is constructed. The process of indoor temperature regulation in the pool is considered.

  8. Crystal engineering of giant molecules based on perylene diimide conjugated polyhedral oligomeric silsesquioxane nano-atom

    Science.gov (United States)

    Ren, He

    Molecular architectures and topologies are found contributing to the formation of supramolecular structures of giant molecules. Dr. Cheng's research group developed a diverse of giant molecules via precisely controlled chemistry synthetic routes. These giant molecules can be categorized into several different families, namely giant surfactants, giant shape amphiphiles and giant polyhedron. By analyzing the hierarchical structures of these carefully designed and precisely synthesized giant molecules, the structural factors which affect, or even dominates, in some cases, the formation of supramolecular structures are revealed in these intensive researches. The results will further contribute to the understanding of dependence of supramolecular structures on molecular designs as well as molecular topology, and providing a practical solution to the scaling up of microscopic molecular functionalities to macroscopic material properties. Molecular Nano Particles (MNPs), including fullerene (C60), POSS, Polyoxometalate (POM) and proteins etc., is defined and applied as a specific type of building blocks in the design and synthesis of giant molecules. The persistence in shape and symmetry is considered as one of the major properties of MNPs. This persistence will support the construction of giant molecules for further supramolecular structures' study by introducing specific shapes, or precisely located side groups which will facilitate self-assembling behaviors with pre-programmed secondary interactions. Dictating material physical properties by its chemical composition is an attractive yet currently failed approach in the study of materials. However, the pursuit of determining material properties by microscopic molecular level properties is never seized, and found its solution when the idea of crystal engineering is raised: should each atom in the material is located exactly where it is designed to be and is properly bonded, the property of the material is hence determined

  9. Eco-engineered rock pools: a concrete solution to biodiversity loss and urban sprawl in the marine environment

    Science.gov (United States)

    Firth, Louise B.; Browne, Keith A.; Knights, Antony M.; Hawkins, Stephen J.; Nash, Róisín

    2016-09-01

    In coastal habitats artificial structures typically support lower biodiversity and can support greater numbers of non-native and opportunistic species than natural rocky reefs. Eco-engineering experiments are typically trialed to succeed; but arguably as much is learnt from failure than from success. Our goal was to trial a generic, cost effective, eco-engineering technique that could be incorporated into rock armouring anywhere in the world. Artificial rock pools were created from manipulated concrete between boulders on the exposed and sheltered sides of a causeway. Experimental treatments were installed in locations where they were expected to fail and compared to controls installed in locations in which they were expected to succeed. Control pools were created lower on the structure where they were immersed on every tidal cycle; experimental pools were created above mean high water spring tide which were only immersed on spring tides. We hypothesised that lower and exposed pools would support significantly higher taxon and functional diversity than upper and sheltered pools. The concrete pools survived the severe winter storms of 2013/14. After 12 months, non-destructive sampling revealed significantly higher mean taxon and functional richness in lower pools than upper pools on the exposed side only. After 24 months the sheltered pools had become inundated with sediments, thus failing to function as rock pools as intended. Destructive sampling on the exposed side revealed significantly higher mean functional richness in lower than upper pools. However, a surprisingly high number of taxa colonised the upper pools leading to no significant difference in mean taxon richness among shore heights. A high number of rare taxa in the lower pools led to total taxon richness being almost twice that of upper pools. These findings highlight that even when expected to fail concrete pools supported diverse assemblages, thus representing an affordable, replicable means of

  10. Fouling behaviors of polybenzimidazole (PBI)-polyhedral oligomeric silsesquioxane (POSS)/polyacrylonitrile (PAN) hollow fiber membranes for engineering osmosis processes

    KAUST Repository

    Chen, Sicong

    2014-02-01

    This paper investigated the individual effects of reverse salt flux and permeate flux on fouling behaviors of as-spun and annealed polybenzimidazole (PBI)-polyhedral oligomeric silsesquioxane (POSS)/polyacrylonitrile (PAN) hollow fiber membranes under forward osmosis (FO) and pressure retarded osmosis (PRO) processes. Two types of membrane fouling had been studied; namely, inorganic fouling (CaSO4·2H2O gypsum scaling) during FO operations and organic fouling (sodium alginate fouling) during PRO operations. It is found that gypsum scaling on the membrane surface may be inhibited and even eliminated with an increase in reverse MgCl2 flux due to competitive formations of MgSO4° and CaSO4·2H2O. In contrast, the increase of reverse NaCl flux exhibits a slight enhancement on alginate fouling in both FO and PRO processes. Comparing to the reverse salt flux, the permeate flux always plays a dominant role in fouling. Therefore, lesser fouling has been observed on the membrane surface under the pressurized PRO process than FO process because the reduced initial flux mitigates the fouling phenomena more significantly than the enhancement caused by an increase in reverse NaCl flux. © 2013 Elsevier B.V.

  11. Software requirements specification for the GIS-T/ISTEA pooled fund study phase C linear referencing engine

    Energy Technology Data Exchange (ETDEWEB)

    Amai, W.; Espinoza, J. Jr. [Sandia National Lab., Albuquerque, NM (United States); Fletcher, D.R. [Univ. of New Mexico, Albuquerque, NM (United States). Alliance for Transportation Research

    1997-06-01

    This Software Requirements Specification (SRS) describes the features to be provided by the software for the GIS-T/ISTEA Pooled Fund Study Phase C Linear Referencing Engine project. This document conforms to the recommendations of IEEE Standard 830-1984, IEEE Guide to Software Requirements Specification (Institute of Electrical and Electronics Engineers, Inc., 1984). The software specified in this SRS is a proof-of-concept implementation of the Linear Referencing Engine as described in the GIS-T/ISTEA pooled Fund Study Phase B Summary, specifically Sheet 13 of the Phase B object model. The software allows an operator to convert between two linear referencing methods and a datum network.

  12. Suppression of pool fires with HRC-125 in a simulated engine nacelle.

    Energy Technology Data Exchange (ETDEWEB)

    Keyser, David R. (INS, Inc., Lexington Park, MD); Hewson, John C.

    2007-06-01

    CFD simulations are conducted to predict the distribution of fire suppressant in an engine nacelle and to predict the suppression of pool fires by the application of this suppressant. In the baseline configuration, which is based on an installed system, suppressant is injected through four nozzles at a rate fast enough to suppress all simulated pool fires. Variations that reduce the mass of the suppression system (reducing the impact of the suppression system on meeting mission needs) are considered, including a reduction in the rate of suppressant injection, a reduction in the mass of suppressant and a reduction in the number of nozzles. In general, these variations should work to reduce the effectiveness of the suppression system, but the CFD results point out certain changes that have negligible impact, at least for the range of phenomena considered here. The results are compared with measurements where available. Comparisons with suppressant measurements are reasonable. A series of twenty-three fire suppression tests were conducted to check the predictions. The pre-test predictions were generally successful in identifying the range of successful suppression tests. In two separate cases, each where one nozzle of the suppression system was capped, the simulation results did indicate a failure to suppress for a condition where the tests indicated successful suppression. When the test-suppressant discharge rate was reduced by roughly 25%, the tests were in agreement with the predictions. That is, the simulations predict a failure to suppress slightly before observed in these cases.

  13. Engineering of charge, biotin-binding and oligomerization of avidin : new tools for avidin-biotin technology

    OpenAIRE

    Marttila, Ari

    2002-01-01

    Avidin is a tetrameric glycoprotein found in chicken egg white. Due to its remarkably high affinity for biotin, avidin has widely been used in biological and biotechnical applications in life sciences. Avidin is also a highly stable protein, maintaining its functional tetrameric structure at high temperatures, extremes of pH, and in the presence of strong denaturants. In the present study we wanted to find out whether different properties of avidin can be modified with protein engineering usi...

  14. Pool boiling performance of Novec{sup TM} 649 engineered fluid

    Energy Technology Data Exchange (ETDEWEB)

    Forrest, Eric; Buongiorno, Jacopo; McKrell, Thomas [Massachusetts Institute of Technology, Cambridge (United States). Dept. of Nuclear Science and Engineering; Hu, Lin-Wen [Massachusetts Institute of Technology, Cambridge (United States). Nuclear Reactor Lab.], e-mail: lwhu@mit.edu

    2009-07-01

    A new fluorinated ketone, C{sub 2}F{sub 5}C(O)CF(CF{sub 3}){sub 2}, is currently being considered as an environmentally friendly alternative for power electronics cooling applications due to its high dielectric strength and low global warming potential (GWP). Sold commercially by the 3M Company as Novec{sup TM} 649 Engineered Fluid, C{sub 2}F{sub 5}C(O)CF(CF{sub 3}){sub 2} exhibits very low acute toxicity while maintaining long-term stability. To assess the general two-phase heat transfer performance of Novec{sup TM} 649, pool boiling tests were conducted by resistively heating a 0.01 in. diameter nickel wire at the fluid's atmospheric saturation temperature of 49 deg C. The nucleate boiling heat transfer coefficient and critical heat flux (CHF) obtained for the fluorinated ketone compare favorably with results obtained for FC-72, a fluorocarbon widely used for the direct cooling of electronic devices. Initial results indicate that Novec{sup TM} 649 may prove to be a viable alternative to FC-72 and other halo alkanes for the cooling of high power density electronic devices. (author)

  15. Using genetic variability available in the breeder's pool to engineer fruit quality.

    Science.gov (United States)

    Orzaez, Diego; Monforte, Antonio J; Granell, Antonio

    2010-01-01

    We substantiate here the opinion that experts in biotechnology and natural biodiversity can work together on the production of successive waves of next-generation GM fruit crops to improve organoleptic and nutritional quality and therefore generate wider public acceptance. In this scenario genetic engineering becomes a faster and more precise way of transferring genes of interest to fruit crop plants from the same or sexually compatible species (intra- or cisgenesis) than more traditional methods, such as MASPB. The availability of complete genome sequences for an increasing number of crop plants, as well as the results from genomics studies, can assist in the identification of gene-to-trait association. The next wave of GM crops will be able to take full advantage of a Synthetic Biology-based strategy in the development of new fruit varieties by using DNA not necessarily present in the breeder's pool for a wide range of applications. There are still a number of challenges which require attention, such as identifying genes and allelic forms associated with traits of interest and improving the precision and stability of the transferred DNA. © 2010 Landes Bioscience

  16. Biodiversity in intertidal rock pools: informing engineering criteria for artificial habitat enhancement in the built environment.

    Science.gov (United States)

    Firth, Louise B; Schofield, Meredith; White, Freya J; Skov, Martin W; Hawkins, Stephen J

    2014-12-01

    Coastal defence structures are proliferating to counter rising and stormier seas. With increasing concern about the ecological value of built environments, efforts are being made to create novel habitat to increase biodiversity. Rock pools are infrequent on artificial structures. We compared biodiversity patterns between rock pools and emergent rock and assessed the role of pool depth and substratum incline in determining patterns of biodiversity. Rock pools were more taxon rich than emergent substrata. Patterns varied with depth and incline with algal groups being more positively associated with shallow than deeper habitats. Substratum incline had little influence on colonising epibiota, with the exception of canopy algae in deeper habitats where vertical surfaces supported greater taxon richness than horizontal surfaces. The creation of artificial rock pools in built environments will have a positive effect on biodiversity. Building pools of varying depths and inclines and shore heights will provide a range of habitats, increase environmental heterogeneity, therefore creating more possible ecological niches, promoting local biodiversity. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Oligomerization paths of the nucleoprotein of influenza A virus.

    Science.gov (United States)

    Tarus, B; Bakowiez, O; Chenavas, S; Duchemin, L; Estrozi, L F; Bourdieu, C; Lejal, N; Bernard, J; Moudjou, M; Chevalier, C; Delmas, B; Ruigrok, R W H; Di Primo, C; Slama-Schwok, A

    2012-03-01

    The influenza viruses contain a segmented, negative strand RNA genome. Each RNA segment is covered by multiple copies of the nucleoprotein (NP) and is associated with the polymerase complex into ribonucleoprotein (RNP) particles. Despite its importance in the virus life cycle, the interactions between the NP and the genome are not well understood. Here, we studied the assembly process of NP-RNA oligomers and analyzed how the oligomeric/monomeric status of RNA-free NP affects RNA binding and oligomerization. Recombinant wild-type NP purified in low salt concentrations and a derived mutant engineered for oligomerization deficiency (R416A) were mainly monomeric in RNA-free solutions as shown by biochemical and electron microscopy techniques. NP monomer formed with RNA a fast 1/1 complex characterized by surface plasmon resonance. In a subsequent and slow process that depended on the RNA length, oligomerization of NP was mediated by RNA binding. In contrast, preparations of wild-type NP purified in high salt concentrations as well as mutant Y148A engineered for deficiency in nucleic acid binding were partly or totally oligomeric in RNA-free solutions. These trimer/tetramer NP oligomers bind directly as oligomers to RNA with a higher affinity than that of the monomers. Both oligomerization routes we characterized could be exploited by cellular or viral factors to modulate or control viral RNA encapsidation by NP. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  18. Metabolic engineering of the purine biosynthetic pathway in Corynebacterium glutamicum results in increased intracellular pool sizes of IMP and hypoxanthine

    Directory of Open Access Journals (Sweden)

    Peifer Susanne

    2012-10-01

    Full Text Available Abstract Background Purine nucleotides exhibit various functions in cellular metabolism. Besides serving as building blocks for nucleic acid synthesis, they participate in signaling pathways and energy metabolism. Further, IMP and GMP represent industrially relevant biotechnological products used as flavor enhancing additives in food industry. Therefore, this work aimed towards the accumulation of IMP applying targeted genetic engineering of Corynebacterium glutamicum. Results Blocking of the degrading reactions towards AMP and GMP lead to a 45-fold increased intracellular IMP pool of 22 μmol gCDW-1. Deletion of the pgi gene encoding glucose 6-phosphate isomerase in combination with the deactivated AMP and GMP generating reactions, however, resulted in significantly decreased IMP pools (13 μmol gCDW-1. Targeted metabolite profiling of the purine biosynthetic pathway further revealed a metabolite shift towards the formation of the corresponding nucleobase hypoxanthine (102 μmol gCDW-1 derived from IMP degradation. Conclusions The purine biosynthetic pathway is strongly interconnected with various parts of the central metabolism and therefore tightly controlled. However, deleting degrading reactions from IMP to AMP and GMP significantly increased intracellular IMP levels. Due to the complexity of this pathway further degradation from IMP to the corresponding nucleobase drastically increased suggesting additional targets for future strain optimization.

  19. Populations, pools, and peccaries: simulating the impact of ecosystem engineers on rainforest frogs

    OpenAIRE

    Ringler, Max; Hödl, Walter; Ringler, Eva

    2015-01-01

    “Ecosystem engineering” describes habitat alteration by an organism that affects another organism; such nontrophic interactions between organisms are a current focus in ecological research. Our study quantifies the actual impact an ecosystem engineer can have on another species by using a previously identified model system—peccaries and rainforest frogs. In a 4-year experiment, we simulated the impact of peccaries on a population of Allobates femoralis (Dendrobatidae) by installing an array o...

  20. Oligomerization in health and disease

    CERN Document Server

    Giraldo, Jesus

    2013-01-01

    This special volume of Progress in Molecular Biology and Translational Science focuses on oligomerization in health and disease. Contributions from leading authorities Informs and updates on all the latest developments in the field.

  1. Biophysical characterization of GPCR oligomerization

    DEFF Research Database (Denmark)

    Mathiasen, Signe

    a quantitative characterization of GPCR oligomerization. The assay provided the first quantification of the association energy of the β2 Adrenergic Receptor (β2AR), a prototypical GPCR. Furthermore we directly observed the time-dependent dimerization of β2AR and Cannabinoid receptor 1 at the single molecule......The biophysical characterization of the fundamental molecular mechanisms behind G-protein coupled receptors (GPCRs) oligomerization is proposed to be paramount for understanding the pharmacological consequence of receptor self-association. Here we developed an in vitro assay that allowed...

  2. Genetic noise control via protein oligomerization

    Energy Technology Data Exchange (ETDEWEB)

    Ghim, C; Almaas, E

    2008-06-12

    general, is of direct implications to organism fitness. Finally, noise control through oligomerization suggests avenues for the design of robust synthetic gene circuits for engineering purposes.

  3. Vernal Pools

    Data.gov (United States)

    California Department of Resources — This is a polygon layer representing existing vernal pool complexes in California's Central Valley, as identified and mapped by Dr. Robert F. Holland. The purpose of...

  4. Engineering

    National Research Council Canada - National Science Library

    Includes papers in the following fields: Aerospace Engineering, Agricultural Engineering, Chemical Engineering, Civil Engineering, Electrical Engineering, Environmental Engineering, Industrial Engineering, Materials Engineering, Mechanical...

  5. Oligomeric proanthocyanidins from mangosteen pericarps.

    Science.gov (United States)

    Fu, Caili; Loo, Alvin Eng Kiat; Chia, Fiona Ping Ping; Huang, Dejian

    2007-09-19

    Oligomeric proanthocyanidins were extracted from mangosteen pericarps and fractionated by a Sephadex LH-20 column to give 0.66% yield (dry matter). (13)C and (1)H NMR signals showed the presence of predominantly procyanidins together with a few prodelphinidin units along with small amounts of stereoisomers of afzelechin/epiafzelechin, catechin/epicatechin, and gallocatechin/epigallocatechin. Depolymerization with benzylmercaptan resulted in epicatechin thioether as the major product, and the mean degree of polymerization was determined to be 6.6. The electron spray ionization-mass spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectra revealed the dominant B type oligomers with mainly epicatechin units and with a small amount of A type oligomers. The isolated proanthocyanidins are potent peroxyl radical scavengers as evidenced by the high oxygen radical scavenging capacity at 1.7 x 10 (4) micromol TE/g, much higher than that of pine bark and grape seed extracts.

  6. Thermodynamic competition between membrane protein oligomeric states

    Science.gov (United States)

    Kahraman, Osman; Haselwandter, Christoph A.

    2016-10-01

    Self-assembly of protein monomers into distinct membrane protein oligomers provides a general mechanism for diversity in the molecular architectures, and resulting biological functions, of membrane proteins. We develop a general physical framework describing the thermodynamic competition between different oligomeric states of membrane proteins. Using the mechanosensitive channel of large conductance as a model system, we show how the dominant oligomeric states of membrane proteins emerge from the interplay of protein concentration in the cell membrane, protein-induced lipid bilayer deformations, and direct monomer-monomer interactions. Our results suggest general physical mechanisms and principles underlying regulation of protein function via control of membrane protein oligomeric state.

  7. Thermodynamic competition between membrane protein oligomeric states

    CERN Document Server

    Kahraman, Osman

    2016-01-01

    Self-assembly of protein monomers into distinct membrane protein oligomers provides a general mechanism for diversity in the molecular architectures, and resulting biological functions, of membrane proteins. We develop a general physical framework describing the thermodynamic competition between different oligomeric states of membrane proteins. Using the mechanosensitive channel of large conductance as a model system, we show how the dominant oligomeric states of membrane proteins emerge from the interplay of protein concentration in the cell membrane, protein-induced lipid bilayer deformations, and direct monomer-monomer interactions. Our results suggest general physical mechanisms and principles underlying regulation of protein function via control of membrane protein oligomeric state.

  8. Espacio Pool

    OpenAIRE

    2013-01-01

    Espacio Pool es un grupo abierto de usuarios vinculados a la Facultad de Bellas Artes UCM que actúa sobre los márgenes que la circundan. Toma su nombre de los estanques vacíos ubicados en los jardines de la cafetería, y elige el término anglosajón por sus connotaciones relativas a lo participativo y autogestionado. Su objetivo es habitar los espacios de “vacuidad” para que sean efectivos y permutadores. Espacio Pool organiza la celebración de encuentros a partir de la construcción de disposit...

  9. Reversible peptide oligomerization over nanoscale gold surfaces

    Directory of Open Access Journals (Sweden)

    Kazushige Yokoyama

    2015-11-01

    Full Text Available A selective oligomeric formation of amyloid beta 1-40 (Ab1-40 monomers over a nanogold colloidal surface was investigated. An unfolded Ab1-40 monomer is considered to construct a dimer or trimer based oligomeric form with its hydrophobic segment placing outward under an acidic condition. Under a basic condition, a conformation of Ab is expected to take a folded monomeric form with its hydrophilic segment folded inward, avoiding the networking with residual colloidal particles. The most probable oligomeric form constructed over a 20 nm gold colloidal surface within a 25 ℃ to 65 ℃ temperature range is a dimer based unit and that over 30 or 40 nm gold colloidal surface below 15 ℃ is concluded to be a trimer based unit. However, selective oligomerization was not successfully reproduced under the rest of the conditions. A dipole-induced dipole interaction must cause a flexible structural change between folded and unfolded forms.

  10. Carbon Nanostructures Containing Polyhedral Oligomeric Silsesquioxanes (POSS)

    NARCIS (Netherlands)

    Potsi, Georgia; Rossos, Andreas; Kouloumpis, Antonios; Antoniou, Myrsini K.; Spyrou, Konstantinos; Karakassides, Michael A.; Gournis, Dimitrios; Rudolf, Petra

    2015-01-01

    This mini review describes the synthesis and properties of carbon nanostructures containing organic-inorganic cage-like polyhedral oligomeric silsesquioxane (POSS). The physical and chemical functionalization of carbon nanomaterials such as graphene, graphene oxide, carbon nanotubes, and fullerenes

  11. Carbon Nanostructures Containing Polyhedral Oligomeric Silsesquioxanes (POSS)

    NARCIS (Netherlands)

    Potsi, Georgia; Rossos, Andreas; Kouloumpis, Antonios; Antoniou, Myrsini K.; Spyrou, Konstantinos; Karakassides, Michael A.; Gournis, Dimitrios; Rudolf, Petra

    2015-01-01

    This mini review describes the synthesis and properties of carbon nanostructures containing organic-inorganic cage-like polyhedral oligomeric silsesquioxane (POSS). The physical and chemical functionalization of carbon nanomaterials such as graphene, graphene oxide, carbon nanotubes, and fullerenes

  12. Dimerization and oligomerization of the chaperone calreticulin

    DEFF Research Database (Denmark)

    Jørgensen, Charlotte S; Ryder, L Rebekka; Steinø, Anne;

    2003-01-01

    protein. Using PAGE, urea gradient gel electrophoresis, capillary electrophoresis and MS, we show that dimerization through the SH group can be induced by lowering the pH to 5-6, heating, or under conditions that favour partial unfolding such as urea concentrations above 2.6 m or SDS concentrations above...... 0.025%. Moreover, we show that calreticulin also has the ability to self-oligomerize through noncovalent interactions at urea concentrations above 2.6 m at pH below 4.6 or above pH 10, at temperatures above 40 degrees C, or in the presence of high concentrations of organic solvents (25%), conditions...... urea or 1% SDS, and heat-induced oligomerization could be inhibited by 8 m urea or 1% SDS when present during heating. Comparison of the binding properties of monomeric and oligomeric calreticulin in solid-phase assays showed increased binding to peptides and denatured proteins when calreticulin...

  13. Oligomeric secoiridoid glucosides from Jasminum abyssinicum.

    Science.gov (United States)

    Gallo, Francesca Romana; Palazzino, Giovanna; Federici, Elena; Iurilli, Raffaella; Monache, Franco Delle; Chifundera, Kusamba; Galeffi, Corrado

    2006-03-01

    From the root bark of Jasminum abyssinicum (Oleaceae) collected in Congo was isolated tree oligomeric secoiridoid glucosides named craigosides A-C. The three compounds are esters of a cyclopentanoid monoterpene with an iridane skeleton, esterified with three, two and two, respectively, units of oleoside 11-methyl ester. The structures were elucidated by spectroscopic methods and chemical correlations.

  14. Genetic noise control via protein oligomerization

    Directory of Open Access Journals (Sweden)

    Almaas Eivind

    2008-11-01

    Full Text Available Abstract Background Gene expression in a cell entails random reaction events occurring over disparate time scales. Thus, molecular noise that often results in phenotypic and population-dynamic consequences sets a fundamental limit to biochemical signaling. While there have been numerous studies correlating the architecture of cellular reaction networks with noise tolerance, only a limited effort has been made to understand the dynamic role of protein-protein interactions. Results We have developed a fully stochastic model for the positive feedback control of a single gene, as well as a pair of genes (toggle switch, integrating quantitative results from previous in vivo and in vitro studies. In particular, we explicitly account for the fast binding-unbinding kinetics among proteins, RNA polymerases, and the promoter/operator sequences of DNA. We find that the overall noise-level is reduced and the frequency content of the noise is dramatically shifted to the physiologically irrelevant high-frequency regime in the presence of protein dimerization. This is independent of the choice of monomer or dimer as transcription factor and persists throughout the multiple model topologies considered. For the toggle switch, we additionally find that the presence of a protein dimer, either homodimer or heterodimer, may significantly reduce its random switching rate. Hence, the dimer promotes the robust function of bistable switches by preventing the uninduced (induced state from randomly being induced (uninduced. Conclusion The specific binding between regulatory proteins provides a buffer that may prevent the propagation of fluctuations in genetic activity. The capacity of the buffer is a non-monotonic function of association-dissociation rates. Since the protein oligomerization per se does not require extra protein components to be expressed, it provides a basis for the rapid control of intrinsic or extrinsic noise. The stabilization of regulatory circuits

  15. Hawaii ESI: POOLS (Anchialine Pool Points)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data set contains sensitive biological resource data for anchialine pools in Hawaii. Anchialine pools are small, relatively shallow coastal ponds that occur...

  16. 某市跳水馆工程标准跳水池专用瓷砖镶贴施工%The special ceramic tile cover construction of a diving hall engineering standard diving pool

    Institute of Scientific and Technical Information of China (English)

    张毅

    2015-01-01

    通过阐述某市跳水馆工程跳水池建设中专用瓷砖面层镶贴项目,从技术和质量角度分析了施工过程中的施工工艺与需要注意的问题,以提高跳水池专用瓷砖的镶贴质量,为专用瓷砖镶贴积累一定的知识和经验。%Through the elaboration of special ceramic tile cover layer project in a city diving hall engineering diving pool construction,from the point of view of technology and quality analyzed the construction process and matters need to pay attention to in construction technology,in order to improve the cover quality of diving pool special ceramic tile,accumulated some experience and knowledge for special ceramic tile cover.

  17. Simulations of Oligomeric Intermediates in Prion Diseases

    CERN Document Server

    Mobley, D L; Singh, R R P; Kulkarni, R V; Slepoy, A; Mobley, David L.; Cox, Daniel L.; Singh, Rajiv R. P.; Kulkarni, Rahul V.; Slepoy, Alexander

    2003-01-01

    We extend our previous stochastic cellular automata based model for areal aggregation of prion proteins on neuronal surfaces. The new anisotropic model allow us to simulate both strong beta-sheet and weaker attachment bonds between proteins. Constraining binding directions allows us to generate aggregate structures with the hexagonal lattice symmetry found in recently observed in vitro experiments. We argue that these constraints on rules may correspond to underlying steric constraints on the aggregation process. We find that monomer dominated growth of the areal aggregate is too slow to account for some observed doubling time-to-incubation time ratios inferred from data, and so consider aggregation dominated by relatively stable but non-infectious oligomeric intermediates. We compare a kinetic theory analysis of oligomeric aggregation to spatially explicit simulations of the process. We find that with suitable rules for misfolding of oligomers, possibly due to water exclusion by the surrounding aggregate, th...

  18. Metal Catalyzed Oligomerization Reactions of Organosiloxanes.

    Science.gov (United States)

    1982-10-28

    Table I. Some monosilanes , e.g. Me3SiH and PhMe2SiH, are found, but none of the expected PhMeSiH 2 is present. However, some of the expected oligomeric...of the monosilanes as indicated in Table I suggested that reaction 17 does occur, but the initially produced PhMeSiH 2 undergoes secondary

  19. Uncertainty analysis of suppression pool heating during an ATWS in a BWR-5 plant. An application of the CSAU methodology using the BNL engineering plant analyzer

    Energy Technology Data Exchange (ETDEWEB)

    Wulff, W.; Cheng, H.S.; Mallen, A.N. [Brookhaven National Lab., Upton, NY (United States); Johnsen, G.W. [Idaho National Engineering Lab., Idaho Falls, ID (United States); Lellouche, G.S. [Technical Data Services, Chicago, IL (United States)

    1994-03-01

    The uncertainty has been estimated of predicting the peak temperature in the suppression pool of a BWR power plant, which undergoes an NRC-postulated Anticipated Transient Without Scram (ATWS). The ATWS is initiated by recirculation-pump trips, and then leads to power and flow oscillations as they had occurred at the LaSalle-2 Power Station in March of 1988. After limit-cycle oscillations have been established, the turbines are tripped, but without MSIV closure, allowing steam discharge through the turbine bypass into the condenser. Postulated operator actions, namely to lower the reactor vessel pressure and the level elevation in the downcomer, are simulated by a robot model which accounts for operator uncertainty. All balance of plant and control systems modeling uncertainties were part of the statistical uncertainty analysis that was patterned after the Code Scaling, Applicability and Uncertainty (CSAU) evaluation methodology. The analysis showed that the predicted suppression-pool peak temperature of 329.3 K (133{degrees}F) has a 95-percentile uncertainty of 14.4 K (26{degrees}F), and that the size of this uncertainty bracket is dominated by the experimental uncertainty of measuring Safety and Relief Valve mass flow rates under critical-flow conditions. The analysis showed also that the probability of exceeding the suppression-pool temperature limit of 352.6 K (175{degrees}F) is most likely zero (it is estimated as < 5-104). The square root of the sum of the squares of all the computed peak pool temperatures is 350.7 K (171.6{degrees}F).

  20. Three-Dimensional Models of the Oligomeric Human Asialoglycoprotein Receptor (ASGP-R

    Directory of Open Access Journals (Sweden)

    Anna Maria Bianucci

    2010-10-01

    Full Text Available The work presented here is aimed at suggesting plausible hypotheses for functional oligomeric forms of the human asialoglycoprotein receptor (ASGP-R, by applying a combination of different computational techniques. The functional ASGP-R is a hetero-oligomer, that comprises of several subunits of two different kinds (H1 and H2, which are highly homologous. Its stoichiometry is still unknown. An articulated step-wise modeling protocol was used in order to build the receptor model in a minimal oligomeric form, necessary for it to bind multi-antennary carbohydrate ligands. The ultimate target of the study is to contribute to increasing the knowledge of interactions between the human ASGP-R and carbohydrate ligands, at the molecular level, pertinent to applications in the field of hepatic tissue engineering.

  1. Polybenzoxazine/Polyhedral Oligomeric Silsesquioxane (POSS Nanocomposites

    Directory of Open Access Journals (Sweden)

    Mohamed Gamal Mohamed

    2016-06-01

    Full Text Available The organic/inorganic hybrid materials from polyhedral oligomeric silsesquioxane (POSS, inorganic nanoparticles and polybenzoxazine (PBZ have received much interesting recently due to their excellent thermal and mechanical properties, flame retardance, low dielectric constant, well-defined inorganic framework at nanosized scale level, and higher performance relative to those of non-hybrid PBZs. This review describes the synthesis, dielectric constants, and thermal, rheological, and mechanical properties of covalently bonded mono- and multifunctionalized benzoxazine POSS hybrids, other functionalized benzoxazine POSS derivatives, and non-covalently (hydrogen bonded benzoxazine POSS composites.

  2. Endocytic pathways mediating oligomeric Aβ42 neurotoxicity

    Directory of Open Access Journals (Sweden)

    Laxton Kevin

    2010-05-01

    Full Text Available Abstract Background One pathological hallmark of Alzheimer's disease (AD is amyloid plaques, composed primarily of amyloid-β peptide (Aβ. Over-production or diminished clearance of the 42 amino acid form of Aβ (Aβ42 in the brain leads to accumulation of soluble Aβ and plaque formation. Soluble oligomeric Aβ (oAβ has recently emerged to be as a likely proximal cause of AD. Results Here we demonstrate that endocytosis is critical in mediating oAβ42-induced neurotoxicity and intraneuronal accumulation of Aβ. Inhibition of clathrin function either with a pharmacological inhibitor, knock-down of clathrin heavy chain expression, or expression of the dominant-negative mutant of clathrin-assembly protein AP180 did not block oAβ42-induced neurotoxicity or intraneuronal accumulation of Aβ. However, inhibition of dynamin and RhoA by expression of dominant negative mutants reduced neurotoxicity and intraneuronal Aβ accumulation. Pharmacologic inhibition of the dynamin-mediated endocytic pathway by genistein also reduced neurotoxicity. Conclusions These data suggest that dynamin-mediated and RhoA-regulated endocytosis are integral steps for oligomeric Aβ42-induced neurotoxicity and intraneuronal Aβ accumulation.

  3. Pooled effect of injection pressure and turbulence inducer piston on performance, combustion, and emission characteristics of a DI diesel engine powered with biodiesel blend.

    Science.gov (United States)

    Isaac JoshuaRamesh Lalvani, J; Parthasarathy, M; Dhinesh, B; Annamalai, K

    2016-12-01

    In this study, the effect of injection pressure on combustion, performance, and emission characteristics of a diesel engine powered with turbulence inducer piston was studied. Engine tests were executed using conventional diesel and 20% blend of adelfa biodiesel [A20]. The results acquired from renewable fuel A20 in the conventional engine showed reduction in brake thermal efficiency being the result of poor air fuel mixing characteristics and the higher viscosity of the tested fuel. This prompted further research aiming at the improvement of turbulence for better air fuel mixing by a novel turbulence inducer piston [TIP]. The investigation was carried out to study the combined effect of injection pressure and turbulence inducer piston. Considerable improvement in the emission characteristics like hydrocarbon, carbon monoxide, smoke was acheived as a result of optimised injection pressure. Nevertheless, the nitrogen oxide emissions were slightly higher than those of the conventional unmodified engine. The engine with turbulence inducer piston shows the scope for reducing the major pollution and thus ensures environmental safety.

  4. Poole High Street study

    OpenAIRE

    Kilburn, David

    2007-01-01

    A presentation given to key decision makers within Poole to improve the retail offer in Poole High Street and leverage the benefit of improved town planning and the introduction of quality retail companies.

  5. Swimming pool cleaner poisoning

    Science.gov (United States)

    Swimming pool cleaner poisoning occurs when someone swallows this type of cleaner, touches it, or breathes in ... The harmful substances in swimming pool cleaner are: Bromine ... copper Chlorine Soda ash Sodium bicarbonate Various mild acids

  6. Swimming Pool Safety

    Science.gov (United States)

    ... Prevention Listen Español Text Size Email Print Share Swimming Pool Safety Page Content ​What is the best way to keep my child safe around swimming pools? An adult should actively watch children at ...

  7. Swimming pool granuloma

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/001357.htm Swimming pool granuloma To use the sharing features on this page, please enable JavaScript. A swimming pool granuloma is a long-term (chronic) skin ...

  8. The science of pooling

    Energy Technology Data Exchange (ETDEWEB)

    Gilbert, E.

    1995-10-01

    The pooling of data from radon studies is described. Pooling refers to the analysis of original data from several studies, not meta-analysis in which summary measures from published data are analyzed. A main objective for pooling is to reduce uncertainty and to obtain more precise estimates of risk than would be available from any single study.

  9. Oligomerization of Uukuniemi virus nucleocapsid protein

    Directory of Open Access Journals (Sweden)

    Katz Anna

    2010-08-01

    Full Text Available Abstract Background Uukuniemi virus (UUKV belongs to the Phlebovirus genus in the family Bunyaviridae. As a non-pathogenic virus for humans UUKV has served as a safe model bunyavirus in a number of studies addressing fundamental questions such as organization and regulation of viral genes, genome replication, structure and assembly. The present study is focused on the oligomerization of the UUKV nucleocapsid (N protein, which plays an important role in several steps of virus replication. The aim was to locate the domains involved in the N protein oligomerization and study the process in detail. Results A set of experiments concentrating on the N- and C-termini of the protein was performed, first by completely or partially deleting putative N-N-interaction domains and then by introducing point mutations of amino acid residues. Mutagenesis strategy was based on the computer modeling of secondary and tertiary structure of the N protein. The N protein mutants were studied in chemical cross-linking, immunofluorescence, mammalian two-hybrid, minigenome, and virus-like particle-forming assays. The data showed that the oligomerization ability of UUKV-N protein depends on the presence of intact α-helices on both termini of the N protein molecule and that a specific structure in the N-terminal region plays a crucial role in the N-N interaction(s. This structure is formed by two α-helices, rich in amino acid residues with aromatic (W7, F10, W19, F27, F31 or long aliphatic (I14, I24 side chains. Furthermore, some of the N-terminal mutations (e.g. I14A, I24A, F31A affected the N protein functionality both in mammalian two-hybrid and minigenome assays. Conclusions UUKV-N protein has ability to form oligomers in chemical cross-linking and mammalian two-hybrid assays. In mutational analysis, some of the introduced single-point mutations abolished the N protein functionality both in mammalian two-hybrid and minigenome assays, suggesting that especially the N

  10. Antioxidant oligomeric proanthocyanidins from Cistus salvifolius.

    Science.gov (United States)

    Qa'Dan, Fadi; Petereit, Frank; Mansoor, Kenza; Nahrstedt, Adolf

    2006-11-01

    The purified proanthocyanidin oligomers of Cistus salvifolius herb extract accounted for 78% of the total proanthocyanidins and 73% of the total antioxidant activity of this extract. To elucidate the structure of the oligomer, it was depolymerized by acid catalysis in the presence of phloroglucinol. The structures of the resulting flavan-3-ols and phloroglucinol adducts were determined on the basis of 1D- and reverse 2D-NMR (HSQC, HMBC) experiments of their peracetylated derivatives, MALDI-TOF-MS and CD spectroscopy. These observations resulting from the degradation with phloroglucinol were confirmed by 13C NMR spectroscopy of the oligomer. The mean molecular weight of the higher oligomeric fraction was estimated to be 5-6 flavan-3-ol-units.

  11. Molecular Plasticity Regulates Oligomerization and Cytotoxicity of the Multipeptide-length Amyloid-beta Peptide Pool

    NARCIS (Netherlands)

    Vandersteen, A.; Masman, M.F.; Baets, G. de; Jonckheere, W.; Werf, K. van der; Marrink, S.J.; Rozenski, J.; Benilova, I.; Strooper, B. de; Subramaniam, V.; Schymkowitz, J.; Rousseau, F.; Broersen, K.

    2012-01-01

    Current therapeutic approaches under development for Alzheimer disease, including gamma-secretase modulating therapy, aim at increasing the production of Abeta(1-38) and Abeta(1-40) at the cost of longer Abeta peptides. Here, we consider the aggregation of Abeta(1-38) and Abeta(1-43) in addition to

  12. Ordering properties of oligomeric columnar discotic liquid crystals

    NARCIS (Netherlands)

    Umesh, C.P.

    2015-01-01

    The synthesis and liquid crystalline ordering properties of oligomeric discotic liquid crystals were investigated. The phase behaviour and surface ordering properties are dependent on among others core type, spacer length and fluorination.    

  13. Cartilage oligomeric matrix protein in patients with juvenile idiopathic arthritis

    DEFF Research Database (Denmark)

    Bjørnhart, Birgitte; Juul, Anders; Nielsen, Susan

    2009-01-01

    Cartilage oligomeric matrix protein (COMP) has been identified as a prognostic marker of progressive joint destruction in rheumatoid arthritis. In this population based study we evaluated associations between plasma concentrations of COMP, disease activity, and growth velocity in patients...

  14. STRUCTURAL BASIS FOR ALLOSTERIC REGULATION OF HUMAN RIBONUCLEOTIDE REDUCTASE BY NUCLEOTIDE-INDUCED OLIGOMERIZATION

    Science.gov (United States)

    Fairman, James Wesley; Wijerathna, Sanath Ranjan; Ahmad, Md. Faiz; Xu, Hai; Nakano, Ryo; Jha, Shalini; Prendergast, Jay; Welin, Martin; Flodin, Susanne; Roos, Annette; Nordlund, Pär; Li, Zongli; Walz, Thomas; Dealwis, Chris Godfrey

    2011-01-01

    Ribonucleotide reductase (RR) is an αnβn (RR1●RR2) complex that maintains balanced dNTP pools by reducing ribonucleoside diphosphates to deoxyribonucleoside diphosphates. RR1 is the catalytic subunit and RR2 houses the free radical required for catalysis. RR is allosterically regulated by its activator ATP and its inhibitor dATP, which regulate RR activity by inducing oligomerization of RR1. Here, we report the first X-ray structures of human RR1 bound to TTP-only, dATP-only, TTP●GDP, TTP●ATP, and TTP●dATP. These structures provide insights into ATP/dATP regulation of RR. At physiological dATP concentrations, RR1 forms inactive hexamers. We determined the first X-ray structure of the RR1●dATP hexamer and used single-particle electron microscopy to visualize the α6●ββ’ 1●dATP holo complex. Site-directed mutagenesis and functional assays confirm that hexamerization is a prerequisite for inhibition by dATP. Our data provide an elegant mechanism for regulating RR activity by dATP-induced oligomerization. PMID:21336276

  15. Hydrodynamic and functional analysis of HIV-1 Vif oligomerization.

    Science.gov (United States)

    Techtmann, Stephen M; Ghirlando, Rodolfo; Kao, Sandra; Strebel, Klaus; Maynard, Ernest L

    2012-03-13

    HIV-1 Vif is an accessory protein that induces the proteasomal degradation of the host restriction factor, apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3G (APOBEC3G). The N-terminal half of Vif binds to APOBEC3G, and the C-terminal half binds to subunits of a cullin 5-based ubiquitin ligase. This Vif-directed ubiquitin ligase induces the degradation of APOBEC3G (a cytidine deaminase) and thereby protects the viral genome from mutation. A conserved PPLP motif near the C-terminus of Vif is essential for Vif function and is also involved in Vif oligomerization. However, the mechanism and functional significance of Vif oligomerization is unclear. We employed analytical ultracentrifugation to examine the oligomeric properties of Vif in solution. Contrary to previous reports, we find that Vif oligomerization does not require the conserved PPLP motif. Instead, our data suggest a more complex mechanism involving interactions among the HCCH motif, the BC box, and downstream residues in Vif. Mutation of residues near the PPLP motif (S165 and V166) affected the oligomeric properties of Vif and weakened the ability of Vif to bind and induce the degradation of APOBEC3G. We propose that Vif oligomerization may represent a mechanism for regulating interactions with APOBEC3G.

  16. 33 CFR 207.170 - Federal Dam, Oklawaha River, Moss Bluff, Fla.; pool level.

    Science.gov (United States)

    2010-07-01

    ... Bluff, Fla.; pool level. 207.170 Section 207.170 Navigation and Navigable Waters CORPS OF ENGINEERS..., Moss Bluff, Fla.; pool level. (a) The level of the pool shall normally be maintained at elevation 56.5 feet above sea level: Provided, That the level of the pool may be raised to not exceeding 58.5...

  17. 33 CFR 207.60 - Federal Dam, Hudson River, Troy, N.Y.; pool level.

    Science.gov (United States)

    2010-07-01

    ..., N.Y.; pool level. 207.60 Section 207.60 Navigation and Navigable Waters CORPS OF ENGINEERS..., N.Y.; pool level. (a) Whenever the elevation of the pool created by the Federal dam at Troy, N.Y... automatically when the pool level rises to an elevation of +18.5 feet mean sea level, and conform in...

  18. Lanthanide Complexes for Oligomerization of Phenyl Isocyanate

    Institute of Scientific and Technical Information of China (English)

    DENG,Ming-Yu; YAO,Ying-Ming; ZHOU,Yu-Fang; ZHANG,Li-Fen; SHEN,Qi

    2003-01-01

    A series of lanthanide complexes including (Ind)3Sm(THF)(1),[(MeCp)2Sm(μ-SPh)(THF)]2(2),[(MeCp)2Y(μ-O-i-Pr)]2(3),(MeCp)3Sm·THF(4),Sm(SPh)3(hmpa)3(5),[(MeCp)2Y-(μ-OCH2CF3)2(6)and (CF3CH2O)3Y(THF)3(7) were synthesized and they have good activity for the oligomerization of phenylisocyanate.Among them 5 shows the highest activity.The conversion is as high as 96.2%,with 1/2500 of the molar ratio of cat./PhNCO.The main components in oligomer were characterized to be a cycdlodimer and a cyclotrimer.The ratio of cyclodimer to cyclotrimer depends on the lanthanide complexes used.7 gave 85.2%cyclotrimer with 1/300 of the molar ratio of cat./PhNCO at 40℃ for 0.5h,while 5 gave 77.6% cyclodimer with 1/300 of the molar ratio of cat./PhNCO at 40℃ for 4h.

  19. Uncoupling of allosteric and oligomeric regulation in a functional hybrid enzyme constructed from Escherichia coli and human ribonucleotide reductase.

    Science.gov (United States)

    Fu, Yuan; Long, Marcus J C; Rigney, Mike; Parvez, Saba; Blessing, William A; Aye, Yimon

    2013-10-08

    An N-terminal-domain (NTD) and adjacent catalytic body (CB) make up subunit-α of ribonucleotide reductase (RNR), the rate-limiting enzyme for de novo dNTP biosynthesis. A strong linkage exists between ligand binding at the NTD and oligomerization-coupled RNR inhibition, inducible by both dATP and nucleotide chemotherapeutics. These observations have distinguished the NTD as an oligomeric regulation domain dictating the assembly of inactive RNR oligomers. Inactive states of RNR differ between eukaryotes and prokaryotes (α6 in human versus α4β4 in Escherichia coli , wherein β is RNR's other subunit); however, the NTD structurally interconnects individual α2 or α2 and β2 dimeric motifs within the respective α6 or α4β4 complexes. To elucidate the influence of NTD ligand binding on RNR allosteric and oligomeric regulation, we engineered a human- E. coli hybrid enzyme (HE) where human-NTD is fused to E. coli -CB. Both the NTD and the CB of the HE bind dATP. The HE specifically partners with E. coli -β to form an active holocomplex. However, although the NTD is the sole physical tether to support α2 and/or β2 associations in the dATP-bound α6 or α4β4 fully inhibited RNR complexes, the binding of dATP to the HE NTD only partially suppresses HE activity and fully precludes formation of higher-order HE oligomers. We postulate that oligomeric regulation is the ultimate mechanism for potent RNR inhibition, requiring species-specific NTD-CB interactions. Such interdomain cooperativity in RNR oligomerization is unexpected from structural studies alone or biochemical studies of point mutants.

  20. PDA: Pooled DNA analyzer

    Directory of Open Access Journals (Sweden)

    Lin Chin-Yu

    2006-04-01

    Full Text Available Abstract Background Association mapping using abundant single nucleotide polymorphisms is a powerful tool for identifying disease susceptibility genes for complex traits and exploring possible genetic diversity. Genotyping large numbers of SNPs individually is performed routinely but is cost prohibitive for large-scale genetic studies. DNA pooling is a reliable and cost-saving alternative genotyping method. However, no software has been developed for complete pooled-DNA analyses, including data standardization, allele frequency estimation, and single/multipoint DNA pooling association tests. This motivated the development of the software, 'PDA' (Pooled DNA Analyzer, to analyze pooled DNA data. Results We develop the software, PDA, for the analysis of pooled-DNA data. PDA is originally implemented with the MATLAB® language, but it can also be executed on a Windows system without installing the MATLAB®. PDA provides estimates of the coefficient of preferential amplification and allele frequency. PDA considers an extended single-point association test, which can compare allele frequencies between two DNA pools constructed under different experimental conditions. Moreover, PDA also provides novel chromosome-wide multipoint association tests based on p-value combinations and a sliding-window concept. This new multipoint testing procedure overcomes a computational bottleneck of conventional haplotype-oriented multipoint methods in DNA pooling analyses and can handle data sets having a large pool size and/or large numbers of polymorphic markers. All of the PDA functions are illustrated in the four bona fide examples. Conclusion PDA is simple to operate and does not require that users have a strong statistical background. The software is available at http://www.ibms.sinica.edu.tw/%7Ecsjfann/first%20flow/pda.htm.

  1. Pool spacing in forest channels

    Science.gov (United States)

    David R. Montgomery; John M. Buffington; Richard D. Smith; Kevin M. Schmidt; George Pess

    1995-01-01

    Field surveys of stream channels in forested mountain drainage basins in southeast Alaska and Washington reveal that pool spacing depends on large woody debris (LWD) loading and channel type, slope, and width. Mean pool spacing in pool-riffle, plane-bed, and forced pool-riffle channels systematically decreases from greater than 13 channel widths per pool to less than 1...

  2. Characterization and preparation of oligomeric procyanidins from Litchi chinensis pericarp.

    Science.gov (United States)

    Sui, Yong; Zheng, Yan; Li, Xiaopeng; Li, Shuyi; Xie, Bijun; Sun, Zhida

    2016-07-01

    The main purpose of this study is to characterize and prepare A-type oligomeric procyanidins from litchi pericarp (Litchi chinensis Baila). The variety of oligomeric procyanidins was characterized by LC-ESI-MS analysis. There were (+)-catechin, (-)-epicatechin, twelve dimers and six trimers of procyanidins were found in litchi pericarp extracts, and A-type procyanidins were much more abundant than B-type procyanidins. The main flavan-3-ol monomer and oligomeric procyanidins in litchi pericarp were (-)-epicatechin, A-type dimers (A1 and A2) and trimer (epicatechin-(4β-8, 2β-O-7)-epicatechin- (4β-8)-epicatechin). Procyanidin A1 (epicatechin-(4β-8, 2β-O-7)-catechin) was identified by NMR in litchi pericarp for the first time. (-)-Epicatechin and oligomeric procyanidins were prepared by the combination of AB-8 column chromatography and Toyopearl HW-40S column chromatography. The results showed that each fraction predominantly owned a single compound and gave a high yield with (-)-epicatechin, A-type dimers (A1 and A2) and trimer, suggesting a useful method to obtain pure (-)-epicatechin and A-type oligomeric procyanidins.

  3. Vitamin D Pooling Project

    Science.gov (United States)

    The Vitamin D Pooling Project of Rarer Cancers brought together investigators from 10 cohorts to conduct a large prospective epidemiologic study of the association between vitamin D status and seven rarer cancers.

  4. Oligomeric viral proteins: small in size, large in presence

    Science.gov (United States)

    Jayaraman, Bhargavi; Smith, Amber M.; Fernandes, Jason D.; Frankel, Alan D.

    2016-01-01

    Viruses are obligate parasites that rely heavily on host cellular processes for replication. The small number of proteins typically encoded by a virus is faced with selection pressures that lead to the evolution of distinctive structural properties, allowing each protein to maintain its function under constraints such as small genome size, high mutation rate, and rapidly changing fitness conditions. One common strategy for this evolution is to utilize small building blocks to generate protein oligomers that assemble in multiple ways, thereby diversifying protein function and regulation. In this review, we discuss specific cases that illustrate how oligomerization is used to generate a single defined functional state, to modulate activity via different oligomeric states, or to generate multiple functional forms via different oligomeric states. PMID:27685368

  5. Cartilage oligomeric matrix protein enhances the vascularization of acellular nerves

    Directory of Open Access Journals (Sweden)

    Wei-ling Cui

    2016-01-01

    Full Text Available Vascularization of acellular nerves has been shown to contribute to nerve bridging. In this study, we used a 10-mm sciatic nerve defect model in rats to determine whether cartilage oligomeric matrix protein enhances the vascularization of injured acellular nerves. The rat nerve defects were treated with acellular nerve grafting (control group alone or acellular nerve grafting combined with intraperitoneal injection of cartilage oligomeric matrix protein (experimental group. As shown through two-dimensional imaging, the vessels began to invade into the acellular nerve graft from both anastomotic ends at day 7 post-operation, and gradually covered the entire graft at day 21. The vascular density, vascular area, and the velocity of revascularization in the experimental group were all higher than those in the control group. These results indicate that cartilage oligomeric matrix protein enhances the vascularization of acellular nerves.

  6. Cartilage oligomeric matrix protein enhances the vascularization of acellular nerves

    Institute of Scientific and Technical Information of China (English)

    Wei-ling Cui; Long-hai Qiu; Jia-yan Lian; Jia-chun Li; Jun Hu; Xiao-lin Liu

    2016-01-01

    Vascularization of acellular nerves has been shown to contribute to nerve bridging. In this study, we used a 10-mm sciatic nerve defect model in rats to determine whether cartilage oligomeric matrix protein enhances the vascularization of injured acellular nerves. The rat nerve defects were treated with acellular nerve grafting (control group) alone or acellular nerve grafting combined with intraperitoneal injection of cartilage oligomeric matrix protein (experimental group). As shown through two-dimensional imaging, the vessels began to invade into the acellular nerve graft from both anastomotic ends at day 7 post-operation, and gradually covered the entire graft at day 21. The vascular density, vascular area, and the velocity of revascularization in the experimental group were all higher than those in the control group. These results indicate that cartilage oligomeric matrix protein enhances the vascularization of acellular nerves.

  7. Crystal Structure of the Marburg Virus VP35 Oligomerization Domain.

    Science.gov (United States)

    Bruhn, Jessica F; Kirchdoerfer, Robert N; Urata, Sarah M; Li, Sheng; Tickle, Ian J; Bricogne, Gérard; Saphire, Erica Ollmann

    2017-01-15

    Marburg virus (MARV) is a highly pathogenic filovirus that is classified in a genus distinct from that of Ebola virus (EBOV) (genera Marburgvirus and Ebolavirus, respectively). Both viruses produce a multifunctional protein termed VP35, which acts as a polymerase cofactor, a viral protein chaperone, and an antagonist of the innate immune response. VP35 contains a central oligomerization domain with a predicted coiled-coil motif. This domain has been shown to be essential for RNA polymerase function. Here we present crystal structures of the MARV VP35 oligomerization domain. These structures and accompanying biophysical characterization suggest that MARV VP35 is a trimer. In contrast, EBOV VP35 is likely a tetramer in solution. Differences in the oligomeric state of this protein may explain mechanistic differences in replication and immune evasion observed for MARV and EBOV.

  8. A Meltwater Pool Discovered in Tianshan Mountains

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    @@ On an expedition to northwest China's Glacier 1 in the TianshanMountains, a research team led by Prof. Li Zhongqin from the CAS Cold and Arid Regions Environmental and Engineering Research Institute found a 15-meter-long, 4-meterhigh ice cliff in the northwest of the source area on the glacier top. To their surprise, the south-facing ice cliff overlooked an approximately 30-m2 pool sprawling on the glacial sheet. The experts say the pool is at least 1.5 m deep and formed by the summer melt.

  9. High Content Analysis of Compositional Heterogeneities to Study GPCR Oligomerization

    DEFF Research Database (Denmark)

    Walsh, Samuel McEwen

    In this thesis I demonstrate how the natural compositional heterogeneities of synthetic and living cell model systems can be used to quantitate the mechanics of G-protein coupled receptor (GPCR) oligomerization with Förster resonance energy transfer (FRET). The thesis is structured around three a...

  10. Cartilage oligomeric matrix protein specific antibodies are pathogenic

    DEFF Research Database (Denmark)

    Geng, Hui; Nandakumar, Kutty Selva; Pramhed, Anna

    2012-01-01

    ABSTRACT: INTRODUCTION: Cartilage oligomeric matrix protein (COMP) is a major non-collagenous component of cartilage. Earlier, we developed a new mouse model for rheumatoid arthritis using COMP. This study was undertaken to investigate the epitope specificity and immunopathogenicity of COMP...

  11. Protic Cationic Oligomeric Ionic Liquids of the Urethane Type

    DEFF Research Database (Denmark)

    Shevchenko, V. V.; Stryutsky, A. V.; Klymenko, N. S.

    2014-01-01

    Protic oligomeric cationic ionic liquids of the oligo(ether urethane) type are synthesized via the reaction of an isocyanate prepolymer based on oligo(oxy ethylene)glycol with M = 1000 with hexamethylene-diisocyanate followed by blocking of the terminal isocyanate groups with the use of amine...

  12. Polyhedral oligomeric silsesquioxane grafted polymer in polymeric foam

    Energy Technology Data Exchange (ETDEWEB)

    King, Bruce A.; Patankar, Kshitish A.; Costeux, Stephane; Jeon, Hyun K.

    2017-01-17

    A polymeric foam article with a polymer matrix defining multiple cells therein has a polymer component with a first polymer that is a polyhedral oligomeric silsesquioxane grafted polymer that has a weight-average molecular weight of two kilograms per mole or higher and 200 kilograms per mole or lower.

  13. Polyhedral oligomeric silsesquioxane grafted polymer in polymeric foam

    Science.gov (United States)

    King, Bruce A.; Patankar, Kshitish A.; Costeux, Stephane; Jeon, Hyun K.

    2017-01-17

    A polymeric foam article with a polymer matrix defining multiple cells therein has a polymer component with a first polymer that is a polyhedral oligomeric silsesquioxane grafted polymer that has a weight-average molecular weight of two kilograms per mole or higher and 200 kilograms per mole or lower.

  14. High Content Analysis of Compositional Heterogeneities to Study GPCR Oligomerization

    DEFF Research Database (Denmark)

    Walsh, Samuel McEwen

    In this thesis I demonstrate how the natural compositional heterogeneities of synthetic and living cell model systems can be used to quantitate the mechanics of G-protein coupled receptor (GPCR) oligomerization with Förster resonance energy transfer (FRET). The thesis is structured around three a...

  15. The Future of Pooling.

    Science.gov (United States)

    Young, Peter C.; Fone, Martin

    1997-01-01

    Discusses seven propositions underlying the strategies that insurance pools can, will, and must pursue: (1) risk management versus risk financing; (2) elimination of windfall advantages; (3) the maintenance of market-dominant status; (4) cost leadership; (5) client focus; (6) innovation and diversification; and (7) leadership challenges. A sidebar…

  16. Income pooling within families

    DEFF Research Database (Denmark)

    Bonke, Jens; Uldall-Poulsen, Hans

    This paper analyses the phenomenon of income-pooling by applying the Danish household expenditure survey, merged with authoritative register information. Responses to additional questions on income sharing among 1696 couples also allows us to analyses whether the intra-household distribution of r...

  17. Crystal Structure of the Marburg Virus VP35 Oligomerization Domain

    Energy Technology Data Exchange (ETDEWEB)

    Bruhn, Jessica F.; Kirchdoerfer, Robert N.; Urata, Sarah M.; Li, Sheng; Tickle, Ian J.; Bricogne, Gérard; Saphire, Erica Ollmann (Scripps); (Globel Phasing); (UCSD)

    2016-11-09

    ABSTRACT

    Marburg virus (MARV) is a highly pathogenic filovirus that is classified in a genus distinct from that of Ebola virus (EBOV) (generaMarburgvirusandEbolavirus, respectively). Both viruses produce a multifunctional protein termed VP35, which acts as a polymerase cofactor, a viral protein chaperone, and an antagonist of the innate immune response. VP35 contains a central oligomerization domain with a predicted coiled-coil motif. This domain has been shown to be essential for RNA polymerase function. Here we present crystal structures of the MARV VP35 oligomerization domain. These structures and accompanying biophysical characterization suggest that MARV VP35 is a trimer. In contrast, EBOV VP35 is likely a tetramer in solution. Differences in the oligomeric state of this protein may explain mechanistic differences in replication and immune evasion observed for MARV and EBOV.

    IMPORTANCEMarburg virus can cause severe disease, with up to 90% human lethality. Its genome is concise, only producing seven proteins. One of the proteins, VP35, is essential for replication of the viral genome and for evasion of host immune responses. VP35 oligomerizes (self-assembles) in order to function, yet the structure by which it assembles has not been visualized. Here we present two crystal structures of this oligomerization domain. In both structures, three copies of VP35 twist about each other to form a coiled coil. This trimeric assembly is in contrast to tetrameric predictions for VP35 of Ebola virus and to known structures of homologous proteins in the measles, mumps, and Nipah viruses. Distinct oligomeric states of the Marburg and Ebola virus VP35 proteins may explain differences between them in polymerase function and immune evasion. These findings may provide a more accurate understanding of the

  18. CERN Electronics Pool presentations

    CERN Multimedia

    2011-01-01

    The CERN Electronics Pool has organised a series of presentations in collaboration with oscilloscope manufacturers. The last one will take place according to the schedule below.   Time will be available at the end of the presentation to discuss your personal needs. The Agilent presentation had to be postponed and will be organised later. -     Lecroy: Thursday, 24 November 2011, in 530-R-030, 14:00 to 16:30.

  19. Preparative isolation of oligomeric procyanidins from Hawthorn (Crataegus spp.).

    Science.gov (United States)

    Zumdick, S; Petereit, F; Luftmann, H; Hensel, A

    2009-04-01

    The oligomeric procyanidins (OPC) from Hawthorn leaves and flowers (Crataegi folium cum flore) are considered to be in part responsible for the cardiotonic clinical activity of the herbal material. Effective methods for rapid isolation of these heterogenous oligomeric clusters with defined molecular weight as reference compounds are not published until now. Therefore the water soluble fraction of an acetone/water (7 + 3) extract of Hawthorn leaves and flowers was fractionated by a combination of MPLC on RP-18 material and preparative HPLC using a diol stationary phase. This procedure resulted in the effective isolation of procyanidins with a distinct degree of polymerization (DP) from dimers DP2 up to tridecamers DP13. Exact mass measurements with negative ESI-TOF/MS were employed to confirm the respective structures of the isolated procyanidins.

  20. Electron Microscopy Structural Insights into CPAP Oligomeric Behavior

    DEFF Research Database (Denmark)

    Alvarez-Cabrera, Ana L; Delgado, Sandra; Gil-Carton, David

    2017-01-01

    that represent obtaining the protein in a soluble, homogeneous state for structural studies. Our work constitutes a systematic structural analysis on multiple oligomers of HsCPAP(897)(-1338), using single-particle electron microscopy (EM) of negatively stained (NS) samples. Based on image classification...... into clearly different regular 3D maps (putatively corresponding to dimers and tetramers) and direct observation of individual images representing other complexes of HsCPAP(897-1338) (i.e., putative flexible monomers and higher-order multimers), we report a dynamic oligomeric behavior of this protein, where...... of atomic models into the NS 3D maps at resolutions around 20 Å is performed only to complement our experimental data, allowing us to hypothesize on the oligomeric composition of the different complexes. In this way, the current EM work represents an initial step toward the structural characterization...

  1. Swimming Pools and Molluscum Contagiosum

    Science.gov (United States)

    ... Travelers’ Health: Smallpox & Other Orthopoxvirus-Associated Infections Poxvirus Swimming Pools Recommend on Facebook Tweet Share Compartir The ... often ask if molluscum virus can spread in swimming pools. There is also concern that it can ...

  2. Pharmacological Profiles of Oligomerized μ-Opioid Receptors

    OpenAIRE

    Ing-Kang Ho; Cynthia Wei-Sheng Lee

    2013-01-01

    Opioids are widely prescribed pain relievers with multiple side effects and potential complications. They produce analgesia via G-protein-protein coupled receptors: μ-, δ-, κ-opioid and opioid receptor-like 1 receptors. Bivalent ligands targeted to the oligomerized opioid receptors might be the key to developing analgesics without undesired side effects and obtaining effective treatment for opioid addicts. In this review we will update the biological effects of μ-opioids on homo- or hetero-ol...

  3. Receptor Oligomerization as a Process Modulating Cellular Semiotics

    DEFF Research Database (Denmark)

    Giorgi, Franco; Bruni, Luis Emilio; Maggio, Roberto

    2010-01-01

    The majority of G protein-coupled receptors (GPCRs) self-assemble in the form dimeric/oligomeric complexes along the plasma membrane. Due to the molecular interactions they participate, GPCRs can potentially provide the framework for discriminating a wide variety of intercellular signals, as based...... and degeneracy may appear as the required feature to integrate the cell system into functional units of progressively higher hierarchical levels....

  4. DNA display of glycoconjugates to emulate oligomeric interactions of glycans

    Directory of Open Access Journals (Sweden)

    Alexandre Novoa

    2015-05-01

    Full Text Available Glycans (carbohydrate portion of glycoproteins and glycolipids frequently exert their function through oligomeric interactions involving multiple carbohydrate units. In efforts to recapitulate the diverse spatial arrangements of the carbohydrate units, assemblies based on hybridization of nucleic acid conjugates have been used to display simplified ligands with tailored interligand distances and valences. The programmability of the assemblies lends itself to a combinatorial display of multiple ligands. Recent efforts in the synthesis and applications of such conjugates are discussed.

  5. Copper-induced oligomerization of peptides: a model study.

    Science.gov (United States)

    Schlosser, Gitta; Stefanescu, Raluca; Przybylski, Michael; Murariu, Manuela; Hudecz, Ferenc; Drochioiu, Gabi

    2007-01-01

    In this work, copper-binding of the tetraglycine peptide (Gly-Gly-Gly-Gly) was studied by electrospray ionization mass spectrometry. Experiments were performed under alkaline conditions, in the presence of ethanolamine (pH 10.95). We observed that the presence of copper(II) ions induces the aggregation of the peptide and the formation of copper-bound complexes with higher molecular mass is favored, such as the oligomer complexes [3M+2Cu-3H](+) and [4M+3Cu-5H](+). At 1:1 peptide-copper(II) ion ratio, the singly charged [3M+2Cu-3H](+) oligomer complex is the base peak in the mass spectrum. Metal ion-induced oligomer-ization of neurotoxic peptides is well known in the literature; however, there are very few examples in which such oligomerization was directly observed by mass spectrometry. Our results show that application of short peptides can be useful to study the -mechanism of metal ion binding and metal ion-induced oligomerization of peptides.

  6. UMRR Pool 18 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  7. UMRR Pool 11 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  8. UMRR Pool 10 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  9. UMRR Pool 08 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  10. UMRR Pool 09 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  11. UMRR Pool 15 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  12. UMRR Pool 19 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  13. UMRR Pool 22 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  14. UMRR Pool 24 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  15. UMRR Pool 20 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  16. UMRR Pool 12 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  17. UMRR Pool 08 Topobathy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Separate data for floodplain elevation and bathymetry were collected on the Upper Mississippi River System (UMRS) by the US Army Corps of Engineers (USACE), Upper...

  18. The Productive Ligurian Pool

    CERN Document Server

    Casella, E; Couvelard, X; Caldeira, R M A

    2011-01-01

    In contrast with the behavior of the eddies in the open-ocean, the sub-mesoscale eddies generated in the constricted Ligurian Basin (NW Mediterranean), are unproductive but their combined effect, arranged in a rim-like fashion, contributes to the containment of a Productive Ligurian Pool (PLP). Data de- rived from MODIS satellite sensor showed persistent higher chlorophyll con- centrations in the centre of the basin, concurrent with high EKE values in its surroundings, derived from AVISO altimetry merged products. This sug- gested that this 'productive pool' is maintained by the intense (sub)mesoscale eddy activity in the rim. Numerical realistic experiments, using a Regional Ocean Model System, forced by MERCATOR and by a high-resolution COSMO- l7 atmospheric model, also showed that most of the sub-mesoscale eddies, during 2009 and 2010, are concentrated in the rim surrounding the basin, contributing to the formation of a basin-scale cyclonic gyre. We hypothesized that the interaction between eddies in the r...

  19. Functional Perfluoroalkyl Polyhedral Oligomeric Silsesquioxane (F-POSS): Building Blocks for Low Surface Energy Materials

    Science.gov (United States)

    2010-10-21

    Technical Paper 3. DATES COVERED (From - To) 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER Functional Perfluoroalkyl Polyhedral Oligomeric Silsesquioxane (F...long chain fluorinated alkyl groups ranging from 6-12 carbon atoms in length. Herein, a disilanol perfluoroalkyl polyhedral oligomeric...FUNCTIONAL PERFLUOROALKYL POLYHEDRAL OLIGOMERIC SILSESQUIOXANES (F-POSS): BUILDING BLOCKS FOR LOW SURFACE ENERGY MATERIA LS Sean M Rami,.e:, Yvonne Dia

  20. hA molecular switch in amyloid assembly: Met35 and amyloid beta-protein oligomerization

    OpenAIRE

    Bitan, Gal; Tarus, Bogdan; Vollers, Sabrina S.; Lashuel, Hilal A.; Condron, Margaret M.; Straub, John E.; Teplow, David B.

    2003-01-01

    Aberrant protein oligomerization is an important pathogenetic process in vivo. In Alzheimer's disease (AD), the amyloid beta-protein (Abeta) forms neurotoxic oligomers. The predominant in vivo Abeta alloforms, Abeta40 and Abeta42, have distinct oligomerization pathways. Abeta42 monomers oligomerize into pentamer/hexamer units (paranuclei) which self-associate to form larger oligomers. Abeta40 does not form these paranuclei, a fact which may explain the particularly strong linkage of Abeta42 w...

  1. Structure-based Design of a Disulfide-lined Oligomeric Form of the Simian Virus 40 (SV40) Large T Antigen DNA-Binding Domain

    Energy Technology Data Exchange (ETDEWEB)

    G Meinke; P Phelan; A Fradet-Turcotte; J Archambault; P Bullock

    2011-12-31

    The modular multifunctional protein large T antigen (T-ag) from simian virus 40 orchestrates many of the events needed for replication of the viral double-stranded DNA genome. This protein assembles into single and double hexamers on specific DNA sequences located at the origin of replication. This complicated process begins when the origin-binding domain of large T antigen (T-ag ODB) binds the GAGGC sequences in the central region (site II) of the viral origin of replication. While many of the functions of purified T-ag OBD can be studied in isolation, it is primarily monomeric in solution and cannot assemble into hexamers. To overcome this limitation, the possibility of engineering intermolecular disulfide bonds in the origin-binding domain which could oligomerize in solution was investigated. A recent crystal structure of the wild-type T-ag OBD showed that this domain forms a left-handed spiral in the crystal with six subunits per turn. Therefore, we analyzed the protein interface of this structure and identified two residues that could potentially support an intermolecular disulfide bond if changed to cysteines. SDS-PAGE analysis established that the mutant T-ag OBD formed higher oligomeric products in a redox-dependent manner. In addition, the 1.7 {angstrom} resolution crystal structure of the engineered disulfide-linked T-ag OBD is reported, which establishes that oligomerization took place in the expected manner.

  2. Photoreceptor engineering

    Directory of Open Access Journals (Sweden)

    Thea eZiegler

    2015-06-01

    Full Text Available Sensory photoreceptors not only control diverse adaptive responses in Nature, but as light-regulated actuators they also provide the foundation for optogenetics, the non-invasive and spatiotemporally precise manipulation of cellular events by light. Novel photoreceptors have been engineered that establish control by light over manifold biological processes previously inaccessible to optogenetic intervention. Recently, photoreceptor engineering has witnessed a rapid development, and light-regulated actuators for the perturbation of a plethora of cellular events are now available. Here, we review fundamental principles of photoreceptors and light-regulated allostery. Photoreceptors dichotomize into associating receptors that alter their oligomeric state as part of light-regulated allostery and non-associating receptors that do not. A survey of engineered photoreceptors pinpoints light-regulated association reactions and order-disorder transitions as particularly powerful and versatile design principles. Photochromic photoreceptors that are bidirectionally toggled by two light colors augur enhanced spatiotemporal resolution and use as photoactivatable fluorophores. By identifying desirable traits in engineered photoreceptors, we provide pointers for the design of future, light-regulated actuators.

  3. Quaternary Structure Analyses of an Essential Oligomeric Enzyme.

    Science.gov (United States)

    Soares da Costa, Tatiana P; Christensen, Janni B; Desbois, Sebastien; Gordon, Shane E; Gupta, Ruchi; Hogan, Campbell J; Nelson, Tao G; Downton, Matthew T; Gardhi, Chamodi K; Abbott, Belinda M; Wagner, John; Panjikar, Santosh; Perugini, Matthew A

    2015-01-01

    Here, we review recent studies aimed at defining the importance of quaternary structure to a model oligomeric enzyme, dihydrodipicolinate synthase. This will illustrate the complementary and synergistic outcomes of coupling the techniques of analytical ultracentrifugation with enzyme kinetics, in vitro mutagenesis, macromolecular crystallography, small angle X-ray scattering, and molecular dynamics simulations, to demonstrate the role of subunit self-association in facilitating protein dynamics and enzyme function. This multitechnique approach has yielded new insights into the molecular evolution of protein quaternary structure.

  4. Spontaneous Oligomerization of Nucleotide Alternatives in Aqueous Solutions

    Science.gov (United States)

    Smith, Karen E.; House, Christopher H.; Dworkin, Jason P.; Callahan, Michael P.

    2017-03-01

    On early Earth, a primitive polymer that could spontaneously form from likely available precursors may have preceded both RNA and DNA as the first genetic material. Here, we report that heated aqueous solutions containing 5-hydroxymethyluracil (HMU) result in oligomers of uracil, heated solutions containing 5-hydroxymethylcytosine (HMC) result in oligomers of cytosine, and heated solutions containing both HMU and HMC result in mixed oligomers of uracil and cytosine. Oligomerization of hydroxymethylated pyrimidines, which may have been abundant on the primitive Earth, might have been important in the development of simple informational polymers.

  5. Morphology of drying blood pools

    Science.gov (United States)

    Laan, Nick; Smith, Fiona; Nicloux, Celine; Brutin, David; D-Blood project Collaboration

    2016-11-01

    Often blood pools are found on crime scenes providing information concerning the events and sequence of events that took place on the scene. However, there is a lack of knowledge concerning the drying dynamics of blood pools. This study focuses on the drying process of blood pools to determine what relevant information can be obtained for the forensic application. We recorded the drying process of blood pools with a camera and measured the weight. We found that the drying process can be separated into five different: coagulation, gelation, rim desiccation, centre desiccation, and final desiccation. Moreover, we found that the weight of the blood pool diminishes similarly and in a reproducible way for blood pools created in various conditions. In addition, we verify that the size of the blood pools is directly related to its volume and the wettability of the surface. Our study clearly shows that blood pools dry in a reproducible fashion. This preliminary work highlights the difficult task that represents blood pool analysis in forensic investigations, and how internal and external parameters influence its dynamics. We conclude that understanding the drying process dynamics would be advancement in timeline reconstitution of events. ANR funded project: D-Blood Project.

  6. Reaction Mechanisms of the Initial Oligomerization of Aluminophosphate.

    Science.gov (United States)

    Xiang, Yan; Xin, Liang; Deetz, Joshua D; Sun, Huai

    2016-05-12

    The mechanisms of aluminophosphate oligomerization were investigated using density functional theory with the SMD solvation model. Two aluminum species, Al(OH)4(-) and Al(H2O)6(3+), and four phosphorus species, H3PO4, H2PO4(-), HPO4(2-), and PO4(3-), were considered as the monomers for polycondensation reactions. It was found that the most favorable pathway to dimerization was a Lewis acid-base reaction: the aprotic oxygen of phosphoric acid (P═O) performs a nucleophilic attack on the central aluminum atom of Al(OH)4(-). Using this mechanism as a pattern, plausible dimerization mechanisms were investigated by varying the proticity and hydration of the phosphorus and aluminum monomers, respectively. The relative reaction rates of each mechanism were estimated under different pH conditions. The chain growth of aluminophosphates to trimers, tetramers, and pentamers and the cyclization of a linear tetramer were also investigated. For oligomerization reactions beyond dimer formation, it is found that cluster growth favors the addition of the phosphoric monomers rather than aluminum monomers.

  7. The oligomeric state of the active Vps4 AAA ATPase

    Science.gov (United States)

    Monroe, Nicole; Han, Han; Gonciarz, Malgorzata D.; Eckert, Debra M.; Karren, Mary Anne; Whitby, Frank G.; Sundquist, Wesley I.; Hill, Christopher P.

    2013-01-01

    The cellular ESCRT pathway drives membrane constriction toward the cytosol and effects membrane fission during cytokinesis, endosomal sorting, and the release of many enveloped viruses, including HIV. A component of this pathway, the AAA ATPase Vps4, provides energy for pathway progression. Although it is established that Vps4 functions as an oligomer, subunit stoichiometry and other fundamental features of the functional enzyme are unclear. Higher-order oligomers have thus far only been characterized for a Walker B mutant of Vps4 in the presence of ATP. Here, we report that although some mutant Vps4 proteins form dodecameric assemblies, active wild-type S. cerevisiae and S. solfataricus Vps4 enzymes can form hexamers in the presence of ATP and ADP, as assayed by size exclusion chromatography and equilibrium analytical ultracentifugation. The Vta1p activator binds hexameric yeast Vps4p without changing the oligomeric state of Vps4p, implying that the active Vta1p:Vps4p complex also contains a single hexameric ring. Additionally, we report crystal structures of two different archaeal Vps4 homologs, whose structures and lattice interactions suggest a conserved mode of oligomerization. Disruption of the proposed hexamerization interface by mutagenesis abolished the ATPase activity of archaeal Vps4 proteins and blocked Vps4p function in S. cerevisiae. These data challenge the prevailing model that active Vps4 is a double ring dodecamer, and argue that, like other type I AAA ATPases, Vps4 functions as a single ring with six subunits. PMID:24161953

  8. Structure-based molecular modeling approaches to GPCR oligomerization.

    Science.gov (United States)

    Kaczor, Agnieszka A; Selent, Jana; Poso, Antti

    2013-01-01

    Classical structure-based drug design techniques using G-protein-coupled receptors (GPCRs) as targets focus nearly exclusively on binding at the orthosteric site of a single receptor. Dimerization and oligomerization of GPCRs, proposed almost 30 years ago, have, however, crucial relevance for drug design. Targeting these complexes selectively or designing small molecules that affect receptor-receptor interactions might provide new opportunities for novel drug discovery. In order to study the mechanisms and dynamics that rule GPCRs oligomerization, it is essential to understand the dynamic process of receptor-receptor association and to identify regions that are suitable for selective drug binding, which may be determined with experimental methods such as Förster resonance energy transfer (FRET) or Bioluminescence resonance energy transfer (BRET) and computational sequence- and structure-based approaches. The aim of this chapter is to provide a comprehensive description of the structure-based molecular modeling methods for studying GPCR dimerization, that is, protein-protein docking, molecular dynamics, normal mode analysis, and electrostatics studies.

  9. Modulation of Innate Immune Signalling by Lipid-Mediated MAVS Transmembrane Domain Oligomerization.

    Directory of Open Access Journals (Sweden)

    Luis Nobre

    Full Text Available RIG-I-like receptors detect viral RNA in infected cells and promote oligomerization of the outer mitochondrial membrane protein MAVS to induce innate immunity to viral infection through type I interferon production. Mitochondrial reactive oxygen species (mROS have been shown to enhance anti-viral MAVS signalling, but the mechanisms have remained obscure. Using a biochemical oligomerization-reporter fused to the transmembrane domain of MAVS, we found that mROS inducers promoted lipid-dependent MAVS transmembrane domain oligomerization in the plane of the outer mitochondrial membrane. These events were mirrored by Sendai virus infection, which similarly induced lipid peroxidation and promoted lipid-dependent MAVS transmembrane domain oligomerization. Our observations point to a role for mROS-induced changes in lipid bilayer properties in modulating antiviral innate signalling by favouring the oligomerization of MAVS transmembrane domain in the outer-mitochondrial membrane.

  10. Effect of leaving group on the oligomerization of 5'-AMP on montmorillonite. [Abstract only

    Science.gov (United States)

    Prabahar, K. Joseph; Ferris, James P.

    1994-01-01

    The oligomerization of imidazole derivative of 5'-AMP (ImpA) in the presence of montmorillonite clay yields oligomers containing up to 10 monomer units. In these reactions, the heterocyclic base, imidazole is the leaving group. In our present study, we synthesized a series of activated nucleotides of 5'AMP using other leaving groups such as pyrazole, 1,2,4-triazole, piperidine, morpholine, 4-aminopyridine, 4-methylaminopyridine, 4-dimethylaminopyridine, 2-aminobenzimidazole etc. to determine the effect of amine leaving group on the products of the oligomerization reaction. Earlier results from our laboratory showed that the presence AppA in the clay reaction of ImpA enhances the oligomerization reaction to yield higher oligomers. We also studied the effect of AppA in the clay mediated oligomerization reaction of the activated nucleotides. Oligomerization of 2-amino-benzimidazole derivative of 5'-AMP gave higher oligomers containing up to nine monomer units in the presence of AppA.

  11. An enigmatic peptide ligation reaction: protease-catalyzed oligomerization of a native protein segment in neat aqueous solution.

    Science.gov (United States)

    Kumaran, S; Datta, D; Roy, R P

    2000-04-01

    We report an enigmatic peptide ligation reaction catalyzed by Glu-specific Staphylococcus aureus V8 protease that occurs in neat aqueous solution around neutral pH utilizing a totally unprotected peptide substrate containing free alpha-carboxyl and alpha-amino groups. V8 protease catalyzed a chain of ligation steps between pH 6 and 8 at 4 degrees C, producing a gamut of covalent oligomers (dimer through octamer or higher) of a native protein segment TAAAKFE (S39) derived from ribonuclease A (RNAse A). Size-exclusion chromatography suggested the absence of strong interaction between the reacting peptides. The circular dichroism spectra of monomer through pentamer showed length-dependent enhancement of secondary structure in the oligomers, suggesting that protease-catalyzed ligation of a monomer to an oligomer resulted in a product that was more structured than its precursor. The relative conformational stability of the oligomers was reflected in their ability to resist proteolysis, indicating that the oligomerization reaction was facilitated as a consequence of the "conformational trapping" of the product. The ligation reaction proceeded in two phases-slow formation and accumulation of the dimer followed by a fast phase of oligomerization, implying that the conformational trap encountered in the oligomerization reaction was a two-step process. The Gly substitution at any position of the TAAAKFE sequence was deleterious, suggesting that the first step of the conformational trap, namely the dimerization reaction, that proceeded very slowly even with the parent peptide, was quite sensitive to amino acid sequence. In contrast, the oligomerization reaction of an Ala analog, AAAAKFE, occurred in much the same way as S39, albeit with faster rate, suggesting that Ala substitution stabilized the overall conformational trapping process. The results suggest the viability of the product-directed "conformational trap" as a mechanism to achieve peptide ligation of totally

  12. 40 CFR 721.3100 - Oligomeric silicic acid ester compound with a hy-droxyl-al-kyla-mine.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Oligomeric silicic acid ester compound... Significant New Uses for Specific Chemical Substances § 721.3100 Oligomeric silicic acid ester compound with a... chemical substance identified generically as oligomeric silicic acid ester compound with...

  13. Rank Pooling for Action Recognition.

    Science.gov (United States)

    Fernando, Basura; Gavves, Efstratios; Oramas M, Jose Oramas; Ghodrati, Amir; Tuytelaars, Tinne

    2017-04-01

    We propose a function-based temporal pooling method that captures the latent structure of the video sequence data - e.g., how frame-level features evolve over time in a video. We show how the parameters of a function that has been fit to the video data can serve as a robust new video representation. As a specific example, we learn a pooling function via ranking machines. By learning to rank the frame-level features of a video in chronological order, we obtain a new representation that captures the video-wide temporal dynamics of a video, suitable for action recognition. Other than ranking functions, we explore different parametric models that could also explain the temporal changes in videos. The proposed functional pooling methods, and rank pooling in particular, is easy to interpret and implement, fast to compute and effective in recognizing a wide variety of actions. We evaluate our method on various benchmarks for generic action, fine-grained action and gesture recognition. Results show that rank pooling brings an absolute improvement of 7-10 average pooling baseline. At the same time, rank pooling is compatible with and complementary to several appearance and local motion based methods and features, such as improved trajectories and deep learning features.

  14. Antioxidant activity of oligomeric acylphloroglucinols from Myrtus communis L.

    Science.gov (United States)

    Rosa, Antonella; Deiana, Monica; Casu, Viviana; Corona, Giulia; Appendino, Giovanni; Bianchi, Federica; Ballero, Mauro; Dessì, M Assunta

    2003-09-01

    The use of myrtle (Myrtus communis L.) as a culinary spice and as a flavoring agent for alcoholic beverages is widespread in the Mediterranean area, and especially in Sardinia. Myrtle contains unique oligomeric non-prenylated acylphloroglucinols, whose antioxidant activity was investigated in various systems. Both semimyrtucommulone (1) and myrtucommulone A (2) showed powerful antioxidant properties, protecting linoleic acid against free radical attack in simple in vitro systems, inhibiting its autoxidation and its FeCl3- and EDTA-mediated oxidation. While both compounds lacked pro-oxidant activity, semimyrtucommulone was more powerful than myrtucommulone A, and was further evaluated in rat liver homogenates for activity against lipid peroxidation induced by ferric-nitrilotriacetate, and in cell cultures for cytotoxicity and the inhibition of TBH- or FeCl3-induced oxidation. The results of these studies established semimyrtucommulone as a novel dietary antioxidant lead.

  15. Green Nanocomposites from Renewable Plant Oils and Polyhedral Oligomeric Silsesquioxanes

    Directory of Open Access Journals (Sweden)

    Takashi Tsujimoto

    2015-06-01

    Full Text Available Green nanocomposites based on renewable plant oils and polyhedral oligomeric silsesquioxanes (POSS have been developed. An acid-catalyzed curing of epoxidized plant oils with oxirane-containing POSS derivatives produced transparent nanocomposite coatings with high gloss surface, in which the organic and inorganic components were linked via covalent bonds. The hardness and mechanical strength were improved by the incorporation of the POSS unit into the organic polymer matrix. Nanostructural analyses of the nanocomposites showed the formation of homogeneous structures at the micrometer scale. On the other hand, such improvements of the coating and mechanical properties were not observed in the composite without covalent bonds between the plant oil-based polymer and POSS unit. The study demonstrates the correlation between the nanostructure of composites and macroscopic properties.

  16. Does oligomerization in fused thiophene affect reactivity and aromaticity?

    Indian Academy of Sciences (India)

    Siddhartha Kr Purkayastha; Pradip Kr Bhattacharyya

    2016-02-01

    Reactivity and aromaticity of a few fused thiophene oligomers and their conformers are discussed in the light of density functional theory (DFT) and conceptual density functional theory. Reactivity parameters, such as hardness () and electrophilicity (), chemical potential () and energy of the HOMO (highest occupied molecular orbital) have been studied. Oligomerization raises the HOMO of the species, which in turn increases the reactivity of the oligomers. The absorption spectra of the species are analysed using TDDFT (time dependent density functional theory). The absorption peaks show red shift with increasing size of the oligomers. Aromaticity of the species is gauged by nucleus independent chemical shift (NICS). The out-of-plane component, (NICSzz) values advocate higher aromatic character at longer distance whereas, NICS supports the reverse.

  17. Cartilage oligomeric matrix protein specific antibodies are pathogenic

    DEFF Research Database (Denmark)

    Geng, Hui; Nandakumar, Kutty Selva; Pramhed, Anna;

    2012-01-01

    ABSTRACT: INTRODUCTION: Cartilage oligomeric matrix protein (COMP) is a major non-collagenous component of cartilage. Earlier, we developed a new mouse model for rheumatoid arthritis using COMP. This study was undertaken to investigate the epitope specificity and immunopathogenicity of COMP...... and the pathogenicity of mAbs was investigated by passive transfer experiments. RESULTS: B cell immunodominant epitopes were localized within 4 antigenic domains of the COMP but with preferential response to the epidermal growth factor (EGF)-like domain. Some of our anti-COMP mAbs showed interactions with the native...... form of COMP, which is present in cartilage and synovium. Passive transfer of COMP-specific mAbs enhanced arthritis when co-administrated with a sub-arthritogenic dose of a mAb specific to collagen type II. Interestingly, we found that a combination of 5 COMP mAbs was capable of inducing arthritis...

  18. Revisiting the oligomerization mechanism of Vibrio cholerae cytolysin, a beta-barrel pore-forming toxin.

    Science.gov (United States)

    Rai, Anand Kumar; Chattopadhyay, Kausik

    2016-06-03

    Vibrio cholerae cytolysin (VCC) is a membrane-damaging beta-barrel pore-forming toxin (beta-PFT). VCC causes permeabilization of the target membranes by forming transmembrane oligomeric beta-barrel pores. Oligomerization is a key step in the mode of action of any beta-PFT, including that of VCC. Earlier studies have identified some of the key residues in VCC that are directly involved in the generation of the inter-protomer contacts, thus playing critical roles in the oligomerization of the membrane-bound toxin. Analysis of the VCC oligomeric pore structure reveals a potential hydrogen-bond network that appears to connect the sidechain of an asparagine residue (Asn582; located within an inter-domain linker sequence) from one protomer to the backbone CO- and NH-groups of the neighbouring protomer, indirectly through water molecules at most of the inter-protomer interfaces. In the present study, we show that the mutation of Asn582Ala affects the oligomerization and the pore-forming activity of VCC in the membrane lipid bilayer of the synthetic lipid vesicles, while the replacement of Asn582Gln results into the restoration of the oligomeric pore-forming ability of the toxin. Using a number of truncated variants of VCC, having deletion in the C-terminal region of the toxin starting from the Asn582 residue or beyond, we also show that the presence of Asn582 is critically required for the oligomerization of the truncated form of the protein.

  19. Effects of Glycine, Water, Ammonia, and Ammonium Bicarbonate on the Oligomerization of Methionine

    Science.gov (United States)

    Huang, Rui; Furukawa, Yoshihiro; Otake, Tsubasa; Kakegawa, Takeshi

    2017-06-01

    The abiotic oligomerization of amino acids may have created primordial, protein-like biological catalysts on the early Earth. Previous studies have proposed and evaluated the potential of diagenesis for the amino acid oligomerization, simulating the formation of peptides that include glycine, alanine, and valine, separately. However, whether such conditions can promote the formation of peptides composed of multiple amino acids remains unclear. Furthermore, the chemistry of pore water in sediments should affect the oligomerization and degradation of amino acids and oligomers, but these effects have not been studied extensively. In this study, we investigated the effects of water, ammonia, ammonium bicarbonate, pH, and glycine on the oligomerization and degradation of methionine under high pressure (150 MPa) and high temperature conditions (175 °C) for 96 h. Methionine is more difficult to oligomerize than glycine and methionine dimer was formed in the incubation of dry powder of methionine. Methionine oligomers as long as trimers, as well as methionylglycine and glycylmethionine, were formed under every condition with these additional compounds. Among the compounds tested, the oligomerization reaction rate was accelerated by the presence of water and by an increase in pH. Ammonia also increased the oligomerization rate but consumed methionine by side reactions and resulted in the rapid degradation of methionine and its peptides. Similarly, glycine accelerated the oligomerization rate of methionine and the degradation of methionine, producing water, ammonia, and bicarbonate through its decomposition. With Gly, heterogeneous dimers (methionylglycine and glycylmethionine) were formed in greater amounts than with other additional compounds although smaller amount of these heterogeneous dimers were formed with other additional compounds. These results suggest that accelerated reaction rates induced by water and co-existing reactive compounds promote the oligomerization

  20. Effects of Glycine, Water, Ammonia, and Ammonium Bicarbonate on the Oligomerization of Methionine

    Science.gov (United States)

    Huang, Rui; Furukawa, Yoshihiro; Otake, Tsubasa; Kakegawa, Takeshi

    2016-09-01

    The abiotic oligomerization of amino acids may have created primordial, protein-like biological catalysts on the early Earth. Previous studies have proposed and evaluated the potential of diagenesis for the amino acid oligomerization, simulating the formation of peptides that include glycine, alanine, and valine, separately. However, whether such conditions can promote the formation of peptides composed of multiple amino acids remains unclear. Furthermore, the chemistry of pore water in sediments should affect the oligomerization and degradation of amino acids and oligomers, but these effects have not been studied extensively. In this study, we investigated the effects of water, ammonia, ammonium bicarbonate, pH, and glycine on the oligomerization and degradation of methionine under high pressure (150 MPa) and high temperature conditions (175 °C) for 96 h. Methionine is more difficult to oligomerize than glycine and methionine dimer was formed in the incubation of dry powder of methionine. Methionine oligomers as long as trimers, as well as methionylglycine and glycylmethionine, were formed under every condition with these additional compounds. Among the compounds tested, the oligomerization reaction rate was accelerated by the presence of water and by an increase in pH. Ammonia also increased the oligomerization rate but consumed methionine by side reactions and resulted in the rapid degradation of methionine and its peptides. Similarly, glycine accelerated the oligomerization rate of methionine and the degradation of methionine, producing water, ammonia, and bicarbonate through its decomposition. With Gly, heterogeneous dimers (methionylglycine and glycylmethionine) were formed in greater amounts than with other additional compounds although smaller amount of these heterogeneous dimers were formed with other additional compounds. These results suggest that accelerated reaction rates induced by water and co-existing reactive compounds promote the oligomerization

  1. Grundfoss: Chlorination of Swimming Pools

    DEFF Research Database (Denmark)

    Hjorth, Poul G.; Hogan, John; Andreassen, Viggo

    1998-01-01

    Grundfos asked for a model, describing the problem of mixing chemicals, being dosed into water systems, to be developed. The application of the model should be dedicated to dosing aqueous solution of chlorine into swimming pools.......Grundfos asked for a model, describing the problem of mixing chemicals, being dosed into water systems, to be developed. The application of the model should be dedicated to dosing aqueous solution of chlorine into swimming pools....

  2. Grundfoss: Chlorination of Swimming Pools

    DEFF Research Database (Denmark)

    Hjorth, Poul G.; Hogan, John; Andreassen, Viggo

    1998-01-01

    Grundfos asked for a model, describing the problem of mixing chemicals, being dosed into water systems, to be developed. The application of the model should be dedicated to dosing aqueous solution of chlorine into swimming pools.......Grundfos asked for a model, describing the problem of mixing chemicals, being dosed into water systems, to be developed. The application of the model should be dedicated to dosing aqueous solution of chlorine into swimming pools....

  3. Optimal Assignment of Cells in C-RAN Deployments with Multiple BBU Pools

    DEFF Research Database (Denmark)

    Holm, Henrik; Checko, Aleksandra; Al-obaidi, Rami;

    2015-01-01

    recommend to divide the area into multiple BBU Pools. In this paper we show how to optimally assign cells to different BBU Pools in such a scenario. By using Integer Linear Programming (ILP) method we derive engineering guidelines for minimizing the CAPital EXpenditure (CAPEX) of C-RAN deployment....

  4. Structure-based design of a disulfide-linked oligomeric form of the simian virus 40 (SV40) large T antigen DNA-binding domain

    Energy Technology Data Exchange (ETDEWEB)

    Meinke, Gretchen; Phelan, Paul [Tufts School of Medicine and the Sackler School of Graduate Biomedical Sciences, 136 Harrison Avenue, Boston, MA 02111 (United States); Fradet-Turcotte, Amélie; Archambault, Jacques [Institut de Recherches Cliniques de Montréal (IRCM), Montreal, Quebec (Canada); Bullock, Peter A., E-mail: peter.bullock@tufts.edu [Tufts School of Medicine and the Sackler School of Graduate Biomedical Sciences, 136 Harrison Avenue, Boston, MA 02111 (United States)

    2011-06-01

    With the aim of forming the ‘lock-washer’ conformation of the origin-binding domain of SV40 large T antigen in solution, using structure-based analysis an intermolecular disulfide bridge was engineered into the origin-binding domain to generate higher order oligomers in solution. The 1.7 Å resolution structure shows that the mutant forms a spiral in the crystal and has the de novo disulfide bond at the protein interface, although structural rearrangements at the interface are observed relative to the wild type. The modular multifunctional protein large T antigen (T-ag) from simian virus 40 orchestrates many of the events needed for replication of the viral double-stranded DNA genome. This protein assembles into single and double hexamers on specific DNA sequences located at the origin of replication. This complicated process begins when the origin-binding domain of large T antigen (T-ag ODB) binds the GAGGC sequences in the central region (site II) of the viral origin of replication. While many of the functions of purified T-ag OBD can be studied in isolation, it is primarily monomeric in solution and cannot assemble into hexamers. To overcome this limitation, the possibility of engineering intermolecular disulfide bonds in the origin-binding domain which could oligomerize in solution was investigated. A recent crystal structure of the wild-type T-ag OBD showed that this domain forms a left-handed spiral in the crystal with six subunits per turn. Therefore, we analyzed the protein interface of this structure and identified two residues that could potentially support an intermolecular disulfide bond if changed to cysteines. SDS–PAGE analysis established that the mutant T-ag OBD formed higher oligomeric products in a redox-dependent manner. In addition, the 1.7 Å resolution crystal structure of the engineered disulfide-linked T-ag OBD is reported, which establishes that oligomerization took place in the expected manner.

  5. 13 CFR 120.1706 - Pool Originator's retained interest in Pool.

    Science.gov (United States)

    2010-01-01

    ... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false Pool Originator's retained interest in Pool. 120.1706 Section 120.1706 Business Credit and Assistance SMALL BUSINESS ADMINISTRATION... Pools § 120.1706 Pool Originator's retained interest in Pool. The Pool Originator must retain...

  6. Transient structural distortion of metal-free Cu/Zn superoxide dismutase triggers aberrant oligomerization

    DEFF Research Database (Denmark)

    Teilum, Kaare; Smith, Melanie H; Schulz, Eike

    2009-01-01

    of the excited-state structure that forms intermolecular contacts in the earliest nonnative dimer/oligomer. The conformational transition that triggers oligomerization is a common feature of WT SOD1 and ALS-associated mutants that have widely different physicochemical properties. But compared with WT SOD1......, the mutants have enhanced structural distortions in their excited states, and in some cases slightly higher excited-state populations and lower kinetic barriers, implying increased susceptibility to oligomerization. Our results provide a unified picture that highlights both (i) a common denominator among......Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease linked to the misfolding of Cu/Zn superoxide dismutase (SOD1). ALS-related defects in SOD1 result in a gain of toxic function that coincides with aberrant oligomerization. The structural events triggering oligomerization have...

  7. Oligomeric protein structure networks: insights into protein-protein interactions

    Directory of Open Access Journals (Sweden)

    Brinda KV

    2005-12-01

    Full Text Available Abstract Background Protein-protein association is essential for a variety of cellular processes and hence a large number of investigations are being carried out to understand the principles of protein-protein interactions. In this study, oligomeric protein structures are viewed from a network perspective to obtain new insights into protein association. Structure graphs of proteins have been constructed from a non-redundant set of protein oligomer crystal structures by considering amino acid residues as nodes and the edges are based on the strength of the non-covalent interactions between the residues. The analysis of such networks has been carried out in terms of amino acid clusters and hubs (highly connected residues with special emphasis to protein interfaces. Results A variety of interactions such as hydrogen bond, salt bridges, aromatic and hydrophobic interactions, which occur at the interfaces are identified in a consolidated manner as amino acid clusters at the interface, from this study. Moreover, the characterization of the highly connected hub-forming residues at the interfaces and their comparison with the hubs from the non-interface regions and the non-hubs in the interface regions show that there is a predominance of charged interactions at the interfaces. Further, strong and weak interfaces are identified on the basis of the interaction strength between amino acid residues and the sizes of the interface clusters, which also show that many protein interfaces are stronger than their monomeric protein cores. The interface strengths evaluated based on the interface clusters and hubs also correlate well with experimentally determined dissociation constants for known complexes. Finally, the interface hubs identified using the present method correlate very well with experimentally determined hotspots in the interfaces of protein complexes obtained from the Alanine Scanning Energetics database (ASEdb. A few predictions of interface hot

  8. Cartilage oligomeric matrix protein in patients with juvenile idiopathic arthritis: relation to growth and disease activity

    DEFF Research Database (Denmark)

    Bjørnhart, Birgitte; Juul, Anders; Nielsen, Susan

    2009-01-01

    OBJECTIVE: Cartilage oligomeric matrix protein (COMP) has been identified as a prognostic marker of progressive joint destruction in rheumatoid arthritis. In this population based study we evaluated associations between plasma concentrations of COMP, disease activity, and growth velocity in patie......OBJECTIVE: Cartilage oligomeric matrix protein (COMP) has been identified as a prognostic marker of progressive joint destruction in rheumatoid arthritis. In this population based study we evaluated associations between plasma concentrations of COMP, disease activity, and growth velocity...

  9. Synaptic vesicle pools and dynamics.

    Science.gov (United States)

    Alabi, AbdulRasheed A; Tsien, Richard W

    2012-08-01

    Synaptic vesicles release neurotransmitter at chemical synapses, thus initiating the flow of information in neural networks. To achieve this, vesicles undergo a dynamic cycle of fusion and retrieval to maintain the structural and functional integrity of the presynaptic terminals in which they reside. Moreover, compelling evidence indicates these vesicles differ in their availability for release and mobilization in response to stimuli, prompting classification into at least three different functional pools. Ongoing studies of the molecular and cellular bases for this heterogeneity attempt to link structure to physiology and clarify how regulation of vesicle pools influences synaptic strength and presynaptic plasticity. We discuss prevailing perspectives on vesicle pools, the role they play in shaping synaptic transmission, and the open questions that challenge current understanding.

  10. Patent pools: Intellectual property rights and competition

    NARCIS (Netherlands)

    Rodriguez, V.

    2010-01-01

    Patent pools do not correct all problems associated with patent thickets. In this respect, patent pools might not stop the outsider problem from striking pools. Moreover, patent pools can be expensive to negotiate, can exclude patent holders with smaller numbers of patents or enable a group of major

  11. Patent pools: Intellectual property rights and competition

    NARCIS (Netherlands)

    Rodriguez, V.

    2010-01-01

    Patent pools do not correct all problems associated with patent thickets. In this respect, patent pools might not stop the outsider problem from striking pools. Moreover, patent pools can be expensive to negotiate, can exclude patent holders with smaller numbers of patents or enable a group of major

  12. Patent pools: Intellectual property rights and competition.

    NARCIS (Netherlands)

    Rodriguez, V.F.

    2010-01-01

    Patent pools do not correct all problems associated with patent thickets. In this respect, patent pools might not stop the outsider problem from striking pools. Moreover, patent pools can be expensive to negotiate, can exclude patent holders with smaller numbers of patents or enable a group of major

  13. 21 CFR 1250.89 - Swimming pools.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Swimming pools. 1250.89 Section 1250.89 Food and... SANITATION Sanitation Facilities and Conditions on Vessels § 1250.89 Swimming pools. (a) Fill and draw swimming pools shall not be installed or used. (b) Swimming pools of the recirculation type shall be...

  14. Controlling Protein Oligomerization with Surface Curvature on the Nanoscale

    Science.gov (United States)

    Kurylowicz, Marty; Dutcher, John

    2011-03-01

    We investigate the effect of surface curvature on the conformation of beta-lactoglobulin (β LG) using Single Molecule Force Spectroscopy. β LG is a model interfacial protein which stabilizes oil droplets in milk and is known to undergo structural rearrangement when adsorbed onto a surface. We reliably control nanoscale surface curvature by creating close-packed monolayers of monodisperse polystyrene (PS) nanoparticles with diameters of 20, 40, 60, 80 and 140 nm, which are stable in aqueous buffer. By adsorbing β LG onto these hydrophobic surfaces and collecting force-extension curves in the fluid phase we can compare the conformation of β LG on 5 different surface curvatures with that on a flat PS film. We demonstrate a transition from oligomeric to monomeric β LG as the surface curvature is increased. Histograms of contour length from fits to peaks in the force-extension curves show a single maximum near 30 nm for β LG adsorbed onto nanoparticles with diameters less than 80 nm. For the larger nanoparticles, the histogram approaches that observed for β LG adsorbed onto a flat PS film, with maxima indicative of β LG dimers and trimers.

  15. Polyhedral Oligomeric Silsesquioxane Functionalized Carbon Dots for Cell Imaging.

    Science.gov (United States)

    Wang, Wen-Jing; Hai, Xin; Mao, Quan-Xing; Chen, Ming-Li; Wang, Jian-Hua

    2015-08-05

    In the present study, octa-aminopropyl polyhedral oligomeric silsesquioxane hydrochloride salt (OA-POSS) functionalized carbon dots (CDs/POSS) are prepared by a one-pot approach with glycerol as carbon source and solvent medium. OA-POSS serves as a passivation agent, and it is obtained via hydrolytic condensation of 3-aminopropyltriethoxysilane (APTES). During the functionalization process, the amino groups on OA-POSS combine with carboxylic groups on the bare CDs via formation of amide bond to construct organic-inorganic hybrid carbon dots. The obtained CDs/POSS are well dispersed in aqueous medium with a diameter of ca. 3.6 nm. It is demonstrated that CDs/POSS provide favorable photoluminescent property with a quantum yield of 24.0%. They also exhibit resistance to photobleaching and excellent photoluminescence stability in the presence of biological sample matrix (characterized by heavy metals and organic molecules), which facilitate cell imaging in biological systems. Both the photoluminescent emission wavelength and the fluorescence intensity depend closely on the excitation wavelength, and thus, it provides a potential for multicolor imaging as demonstrated with HeLa cells and MCF-7 cells.

  16. Accurate length control of supramolecular oligomerization: Vernier assemblies.

    Science.gov (United States)

    Hunter, Christopher A; Tomas, Salvador

    2006-07-12

    Linear oligomeric supramolecular assemblies of defined length have been generated using the Vernier principle. Two molecules, containing a different number (n and m) of mutually complementary binding sites, separated by the same distance, interact with each other to form an assembly of length (n x m). The assembly grows in the same way as simple supramolecular polymers, but at a molecular stop signal, when the binding sites come into register, the assembly terminates giving an oligomer of defined length. This strategy has been realized using tin and zinc porphyrin oligomers as the molecular building blocks. In the presence of isonicotinic acid, a zinc porphyrin trimer and a tin porphyrin dimer form a 3:4 triple stranded Vernier assembly six porphyrins long. The triple strand Vernier architecture introduced here adds an additional level of cooperativity, yielding a stability and selectivity that cannot be achieved via a simple Vernier approach. The assembly properties of the system were characterized using fluorescence titrations and size-exclusion chromatography (SEC). Assembly of the Vernier complex is efficient at micromolar concentrations in nonpolar solvents, and under more competitive conditions, a variety of fragmentation assemblies can be detected, allowing determination of the stability constants for this system and detailed speciation profiles to be constructed.

  17. Does carbon dioxide pool or stream in the subsurface?

    CERN Document Server

    Cardoso, Silvana S S

    2014-01-01

    Pools of carbon dioxide are found in natural geological accumulations and in engineered storage in saline aquifers. It has been thought that once this CO2 dissolves in the formation water, making it denser, convection streams would transport it efficiently to depth, but this may not be so. Here, we assess the impact of natural chemical reactions between the dissolved CO2 and the rock formation on the convection streams in the subsurface. We show that, while in carbonate rocks the streaming of dissolved carbon dioxide persists, the chemical interactions in silicate-rich rocks may curb this transport drastically and even inhibit it altogether. New laboratory experiments confirm the curtailing of convection by reaction. Wide and narrow streams of dense carbon-rich water are shut-off gradually as reaction strength increases until all transport of the pooled carbon dioxide occurs by slow molecular diffusion. These results show that the complex fluid dynamic and kinetic interactions between pooled carbon dioxide an...

  18. EP BICYCLE POOL - VIGNETTES 2002

    CERN Multimedia

    EP-SMI Help Desk

    2002-01-01

    The vignettes (insurance certificates) for 2002 become obligatory from 1 June. If you have a bicycle from the EP Pool, please bring it to the EP-SMI Help Desk (Building 124) on any working day up to 31 May between 8h.30 - 12h.00 or 13h.30 - 17h.30. EP-SMI Help Desk

  19. Oligomerization of optically active N-(4-hydroxyphenylmandelamide in the presence of β-cyclodextrin and the minor role of chirality

    Directory of Open Access Journals (Sweden)

    Helmut Ritter

    2014-10-01

    Full Text Available The oxidative oligomerization of a chiral mandelamide derivative (N-(4-hydroxyphenylmandelamide, 1 was performed in the presence of horseradish peroxidase, laccase and N,N'-bis(salicylideneethylenediamine-iron(II to obtain chiral oligophenols 2. The low enantioselectivity of the enzymatic catalyzed asymmetric enantiomer-differentiating oligomerizations was investigated. In addition, the poor influence of cyclodextrin on the enantioselectivity of enzymatic catalyzed asymmetric enantiomer-differentiating oligomerizations was studied.

  20. Binding and Oligomerization of Modified and Native Bt Toxins in Resistant and Susceptible Pink Bollworm.

    Directory of Open Access Journals (Sweden)

    Josue Ocelotl

    Full Text Available Insecticidal proteins from Bacillus thuringiensis (Bt are used extensively in sprays and transgenic crops for pest control, but their efficacy is reduced when pests evolve resistance. Better understanding of the mode of action of Bt toxins and the mechanisms of insect resistance is needed to enhance the durability of these important alternatives to conventional insecticides. Mode of action models agree that binding of Bt toxins to midgut proteins such as cadherin is essential for toxicity, but some details remain unresolved, such as the role of toxin oligomers. In this study, we evaluated how Bt toxin Cry1Ac and its genetically engineered counterpart Cry1AcMod interact with brush border membrane vesicles (BBMV from resistant and susceptible larvae of Pectinophora gossypiella (pink bollworm, a global pest of cotton. Compared with Cry1Ac, Cry1AcMod lacks 56 amino acids at the amino-terminus including helix α-1; previous work showed that Cry1AcMod formed oligomers in vitro without cadherin and killed P. gossypiella larvae harboring cadherin mutations linked with >1000-fold resistance to Cry1Ac. Here we found that resistance to Cry1Ac was associated with reduced oligomer formation and insertion. In contrast, Cry1AcMod formed oligomers in BBMV from resistant larvae. These results confirm the role of cadherin in oligomerization of Cry1Ac in susceptible larvae and imply that forming oligomers without cadherin promotes toxicity of Cry1AcMod against resistant P. gossypiella larvae that have cadherin mutations.

  1. Characterization of tau oligomeric seeds in progressive supranuclear palsy.

    Science.gov (United States)

    Gerson, Julia E; Sengupta, Urmi; Lasagna-Reeves, Cristian A; Guerrero-Muñoz, Marcos J; Troncoso, Juan; Kayed, Rakez

    2014-06-14

    Progressive supranuclear palsy (PSP) is a neurodegenerative tauopathy which is primarily defined by the deposition of tau into globose-type neurofibrillary tangles (NFT). Tau in its native form has important functions for microtubule dynamics. Tau undergoes alternative splicing in exons 2, 3, and 10 which results in six different isoforms. Products of splicing on exon 10 are the most prone to mutations. Three repeat (3R) and four repeat (4R) tau, like other disease-associated amyloids, can form oligomers which may then go on to further aggregate and form fibrils. Recent studies from our laboratory and others have provided evidence that tau oligomers, not NFTs, are the most toxic species in neurodegenerative tauopathies and seed the pathological spread of tau. Analysis of PSP brain sections revealed globose-type NFTs, as well as both phosphorylated and unphosphorylated tau oligomers. Analysis of PSP brains via Western blot and ELISA revealed the presence of increased levels of tau oligomers compared to age-matched control brains. Oligomers were immunoprecipitated from PSP brain and were capable of seeding the oligomerization of both 3R and 4R tau isoforms. This is the first time tau oligomers have been characterized in PSP. These results indicate that tau oligomers are an important component of PSP pathology, along with NFTs. The ability of PSP brain-derived tau oligomers to seed 3R and 4R tau suggests that these oligomers represent the pathological species responsible for disease propagation and the presence of oligomers in a pure neurodegenerative tauopathy implies a common neuropathological process for tau seen in diseases with other amyloid proteins.

  2. Interfacial behavior of cytoplasmic and mitochondrial creatine kinase oligomeric states.

    Science.gov (United States)

    Vernoux, Nathalie; Granjon, Thierry; Marcillat, Olivier; Besson, Françoise; Vial, Christian

    2006-03-01

    Adsorption to the air/water interface of isoenzymes of creatine kinase was investigated using surface pressure-area isotherms and Brewster angle microscopy (BAM) observations. Octameric mitochondrial creatine kinase (mtCK) exhibits a significant affinity for the air/water interface. Whatever the mode of formation of the interfacial film, i.e., injection of the protein in the subphase or spreading onto the buffer surface, the final arrangement and conformation adopted by mtCK molecules lead to a similar result. In contrast, the dimeric isoenzymes mtCK and cytosolic MMCK do not induce any surface pressure variation. However, when the subphase contains 0.3M NaCl, both isoenzymes adsorb to the interface. When treated with 0.8 or 3M GdnHCl, muscle creatine kinase (MMCK) becomes surface active and occupies a greater surface than mtCK. This result contrasts with previous observations, often derived from monomeric proteins, that their surface activity is increased upon unfolding. It underlines the possible influence exerted by the protein oligomeric state on its interfacial activity. At a subphase pH of 8.8, which corresponds to the pI of octameric mtCK, the profiles of the isotherms obtained with dimeric and octameric states and the resistance to compression of the protein monolayers are significantly affected when compared to those recorded at pH 7.4. These data suggest that the octamer is more hydrophobic than the dimer and may contribute to explaining why octamers bind to the inner mitochondrial membrane while dimers do not. Copyright 2005 Wiley Periodicals, Inc.

  3. [Infections transmitted in swimming pools].

    Science.gov (United States)

    von Suzani, C; Hazeghi, P

    1976-01-01

    Public swimmingpools can be the source of infections due to micro-organism such as mycobacterium balnei, adeno and enteroviruses, the virus of plantar warts and molluscum contagiosum, the TRIC-Agent of swimmingpool-conjonctivitis and pathogenic fungi. The transmission of trichomonas vaginalis is considered unlikely-Water of pools, supposed to present satisfactory qualities by standard controls, was found to contain pathogenic staphylococci and pseudomonas aeruginosa. Effective preventive measures include the continuous recording of the redox-potential of the water, limiting the number of visitors to pool design specifications, better desinfection of sanitary installations, regular maintenance of technical equipment including frequent backwashing of filters and exclusion of visitors with communicable disease.

  4. A Consensual Linear Opinion Pool

    CERN Document Server

    Carvalho, Arthur

    2012-01-01

    We propose a pooling method to aggregate expert opinions. Intuitively, it works as if the experts were continuously updating their opinions in order to accommodate the expertise of others. Each updated opinion takes the form of a linear opinion pool, where the weight that an expert assigns to a peer's opinion is inversely related to the distance between their opinions. In other words, experts are assumed to prefer opinions that are close to their own opinions. We prove that such an updating process leads to consensus, i.e., the experts all converge towards the same opinion. Further, we show that if experts are rewarded using the quadratic scoring rule, then the above mentioned assumption follows naturally. We empirically demonstrate the efficacy of our method using real-world data.

  5. Bax phosphorylation association with nucleus and oligomerization after neonatal hypoxia-ischemia.

    Science.gov (United States)

    Infante, Smitha Krishna; Oberhauser, Andres F; Perez-Polo, J Regino

    2013-09-01

    Neonatal hypoxia-ischemia (HI) is a common occurrence in preterm and low-birth-weight infants, and the incidence of low-birth-weight and preterm births is increasing. Characterization of brain injury after HI is of critical importance in developing new treatments that more accurately target the injury. After severe HI, neuronal cells undergo necrosis and secondary apoptosis of the surrounding cells as a result of neuroinflammation. We sought to characterize the biochemical pathways associated with cell death after HI. Bax, a cell death signaling protein, is activated after HI and translocates to the nucleus, endoplasmic reticulum, and mitochondria. The translocation patterns of Bax affect the resultant cell death phenotype (necrotic or apoptotic) observed. Although Bax is known to oligomerize once it is activated, less is known about the factors that control its translocation and oligomerization. We hypothesize that Bax kinase-specific phosphorylation determines its oligomerization and intracellular localization. Using well-established in vivo and in vitro models of neonatal HI, we characterized Bax oligomerization and multiorganelle translocation. We found that HI-dependent phosphorylation of Bax determines its oligomerization status and multiorganelle localization, and, ultimately, the cell death phenotype observed. Understanding the mechanisms of Bax translocation will aid in the rational design of therapeutic strategies that decrease the trauma resulting from HI-associated inflammation.

  6. Tris-acetate polyacrylamide gradient gel electrophoresis for the analysis of protein oligomerization.

    Science.gov (United States)

    Cubillos-Rojas, Monica; Schneider, Taiane; Sánchez-Tena, Susana; Bartrons, Ramon; Ventura, Francesc; Rosa, Jose Luis

    2016-02-01

    Here we report a new approach for studying protein oligomerization in cells using a single electrophoresis gel. We combined the use of a crosslinking reagent for sample preparation, such as glutaraldehyde, with the analysis of oligomers by Tris-acetate polyacrylamide gel electrophoresis. The use of a 3-15% Tris-acetate polyacrylamide gradient gel allows for the simultaneous analysis of proteins of masses ranging from 10 to 500 kDa. We showed the usefulness of this method for analyzing endogenous p53 oligomerization with high resolution and sensitivity in human cells. Oligomerization analysis was dependent on the crosslinker concentration used. We also showed that this method could be used to study the regulation of oligomerization. In all experiments, Tris-acetate polyacrylamide gel electrophoresis proved to be a robust, manageable, and cost- and time-efficient method that provided excellent results using a single gel. This approach can be easily extrapolated to the study of other oligomers. All of these features make this method a highly useful tool for the analysis of protein oligomerization.

  7. A fluorimetric readout reporting the kinetics of nucleotide-induced human ribonucleotide reductase oligomerization.

    Science.gov (United States)

    Fu, Yuan; Lin, Hong-Yu; Wisitpitthaya, Somsinee; Blessing, William A; Aye, Yimon

    2014-11-24

    Human ribonucleotide reductase (hRNR) is a target of nucleotide chemotherapeutics in clinical use. The nucleotide-induced oligomeric regulation of hRNR subunit α is increasingly being recognized as an innate and drug-relevant mechanism for enzyme activity modulation. In the presence of negative feedback inhibitor dATP and leukemia drug clofarabine nucleotides, hRNR-α assembles into catalytically inert hexameric complexes, whereas nucleotide effectors that govern substrate specificity typically trigger α-dimerization. Currently, both knowledge of and tools to interrogate the oligomeric assembly pathway of RNR in any species in real time are lacking. We therefore developed a fluorimetric assay that reliably reports on oligomeric state changes of α with high sensitivity. The oligomerization-directed fluorescence quenching of hRNR-α, covalently labeled with two fluorophores, allows for direct readout of hRNR dimeric and hexameric states. We applied the newly developed platform to reveal the timescales of α self-assembly, driven by the feedback regulator dATP. This information is currently unavailable, despite the pharmaceutical relevance of hRNR oligomeric regulation.

  8. Biocatalytically Oligomerized Epicatechin with Potent and Specific Anti-proliferative Activity for Human Breast Cancer Cells

    Directory of Open Access Journals (Sweden)

    Ramaswamy Nagarajan

    2008-11-01

    Full Text Available Catechins, naturally occurring flavonoids derived from wine and green tea, are known to exhibit multiple health benefits. Epigallocatechin gallate (EGCG is one of the most widely investigated catechins, but its efficacy in cancer therapy is still inconsistent and limited. The poor stability of EGCG has contributed to the disparity in the reported anti-cancer activity and other beneficial properties. Here we report an innovative enzymatic strategy for the oligomerization of catechins (specifically epicatechin that yields stable, water-soluble oligomerized epicatechins with enhanced and highly specific anti-proliferative activity for human breast cancer cells. This one-pot oxidative oligomerization is carried out in ambient conditions using Horseradish Peroxidase (HRP as a catalyst yielding water-soluble oligo(epicatechins. The oligomerized epicatechins obtained exhibit excellent growth inhibitory effects against human breast cancer cells with greater specificity towards growth-inhibiting cancer cells as opposed to normal cells, achieving a high therapeutic differential. Our studies indicate that water-soluble oligomeric epicatechins surpass EGCG in stability, selectivity and efficacy at lower doses.

  9. Sustainability of common pool resources

    Science.gov (United States)

    Timilsina, Raja Rajendra; Kamijo, Yoshio

    2017-01-01

    Sustainability has become a key issue in managing natural resources together with growing concerns for capitalism, environmental and resource problems. We hypothesize that the ongoing modernization of competitive societies, which we refer to as “capitalism,” affects human nature for utilizing common pool resources, thus compromising sustainability. To test this hypothesis, we design and implement a set of dynamic common pool resource games and experiments in the following two types of Nepalese areas: (i) rural (non-capitalistic) and (ii) urban (capitalistic) areas. We find that a proportion of prosocial individuals in urban areas is lower than that in rural areas, and urban residents deplete resources more quickly than rural residents. The composition of proself and prosocial individuals in a group and the degree of capitalism are crucial in that an increase in prosocial members in a group and the rural dummy positively affect resource sustainability by 65% and 63%, respectively. Overall, this paper shows that when societies move toward more capitalistic environments, the sustainability of common pool resources tends to decrease with the changes in individual preferences, social norms, customs and views to others through human interactions. This result implies that individuals may be losing their coordination abilities for social dilemmas of resource sustainability in capitalistic societies. PMID:28212426

  10. Sustainability of common pool resources.

    Science.gov (United States)

    Timilsina, Raja Rajendra; Kotani, Koji; Kamijo, Yoshio

    2017-01-01

    Sustainability has become a key issue in managing natural resources together with growing concerns for capitalism, environmental and resource problems. We hypothesize that the ongoing modernization of competitive societies, which we refer to as "capitalism," affects human nature for utilizing common pool resources, thus compromising sustainability. To test this hypothesis, we design and implement a set of dynamic common pool resource games and experiments in the following two types of Nepalese areas: (i) rural (non-capitalistic) and (ii) urban (capitalistic) areas. We find that a proportion of prosocial individuals in urban areas is lower than that in rural areas, and urban residents deplete resources more quickly than rural residents. The composition of proself and prosocial individuals in a group and the degree of capitalism are crucial in that an increase in prosocial members in a group and the rural dummy positively affect resource sustainability by 65% and 63%, respectively. Overall, this paper shows that when societies move toward more capitalistic environments, the sustainability of common pool resources tends to decrease with the changes in individual preferences, social norms, customs and views to others through human interactions. This result implies that individuals may be losing their coordination abilities for social dilemmas of resource sustainability in capitalistic societies.

  11. Bioreactor scale up and protein product quality characterization of piggyBac transposon derived CHO pools.

    Science.gov (United States)

    Rajendra, Yashas; Balasubramanian, Sowmya; Peery, Robert B; Swartling, James R; McCracken, Neil A; Norris, Dawn L; Frye, Christopher C; Barnard, Gavin C

    2017-03-01

    Chinese hamster ovary (CHO) cells remain the most popular host for the production of biopharmaceutical drugs, particularly monoclonal antibodies (mAbs), bispecific antibodies, and Fc-fusion proteins. Creating and characterizing the stable CHO clonally-derived cell lines (CDCLs) needed to manufacture these therapeutic proteins is a lengthy and laborious process. Therefore, CHO pools have increasingly been used to rapidly produce protein to support and enable preclinical drug development. We recently described the generation of CHO pools yielding mAb titers as high as 7.6 g/L in a 16 day bioprocess using piggyBac transposon-mediated gene integration. In this study, we wanted to understand why the piggyBac pool titers were significantly higher (2-10 fold) than the control CHO pools. Higher titers were the result of a combination of increased average gene copy number, significantly higher messenger RNA levels and the homogeneity (i.e. less diverse population distribution) of the piggyBac pools, relative to the control pools. In order to validate the use of piggyBac pools to support preclinical drug development, we then performed an in-depth product quality analysis of purified protein. The product quality of protein obtained from the piggyBac pools was very similar to the product quality profile of protein obtained from the control pools. Finally, we demonstrated the scalability of these pools from shake flasks to 36L bioreactors. Overall, these results suggest that gram quantities of therapeutic protein can be rapidly obtained from piggyBac CHO pools without significantly changing product quality attributes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:534-540, 2017. © 2017 American Institute of Chemical Engineers.

  12. Essential vernal pool habitat action plan

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Vernal pool ecosystem conservation and recovery requires the recovery team to develop methods to determine the distribution of vernal pool types throughout the Great...

  13. Pool power control in remelting systems

    Science.gov (United States)

    Williamson, Rodney L.; Melgaard, David K.; Beaman, Joseph J.

    2011-12-13

    An apparatus for and method of controlling a remelting furnace comprising adjusting current supplied to an electrode based upon a predetermined pool power reference value and adjusting the electrode drive speed based upon the predetermined pool power reference value.

  14. CDC Study Finds Fecal Contamination in Pools

    Science.gov (United States)

    ... Communication (404) 639-3286 CDC study finds fecal contamination in pools A study of public pools done ... The E. coli is a marker for fecal contamination. Finding a high percentage of E. coli-positive ...

  15. Interleukin-1 beta converting enzyme requires oligomerization for activity of processed forms in vivo.

    Science.gov (United States)

    Gu, Y; Wu, J; Faucheu, C; Lalanne, J L; Diu, A; Livingston, D J; Su, M S

    1995-05-01

    Interleukin-1 beta converting enzyme (ICE) is composed of 10' (p10) and 20 kDa (p20) subunits, which are derived from a common 45 kDa precursor. Recent crystallographic studies have shown that ICE exists as a tetramer (p20/p10)2 in the crystal lattice. We provide evidence that the p10 and p20 subunits of ICE associate as oligomers in transfected COS cells. Using intragenic complementation, we show that the activity of a p10/p10 interface mutant defective in autoprocessing can be restored by co-expression with active site ICE mutants. Different active site mutants can also complement each other by oligomerization to form active ICE. These studies indicate that ICE precursor polypeptides may associate in different quaternary structures and that oligomerization is required for autoprocessing. Furthermore, integenic complementation of active site mutants of ICE and an ICE homolog restores autoprocessing activity, suggesting that hetero-oligomerization occurs between ICE homologs.

  16. N-(1-Pyrenyl Maleimide Induces Bak Oligomerization and Mitochondrial Dysfunction in Jurkat Cells

    Directory of Open Access Journals (Sweden)

    Pei-Rong Huang

    2015-01-01

    Full Text Available N-(1-pyrenyl maleimide (NPM is a fluorescent reagent that is frequently used as a derivatization agent for the detection of thio-containing compounds. NPM has been shown to display a great differential cytotoxicity against hematopoietic cancer cells. In this study, the molecular mechanism by which NPM induces apoptosis was examined. Here, we show that treatment of Jurkat cells with NPM leads to Bak oligomerization, loss of mitochondrial membrane potential (Δψm, and release of cytochrome C from mitochondria to cytosol. Induction of Bak oligomerization appears to play a critical role in NPM-induced apoptosis, as downregulation of Bak by shRNA significantly prevented NPM-induced apoptosis. Inhibition of caspase 8 by Z-IETD-FMK and/or depletion of Bid did not affect NPM-induced oligomerization of Bak. Taken together, these results suggest that NPM-induced apoptosis is mediated through a pathway that is independent of caspase-8 activation.

  17. Reactive bay functionalized perylene monoimide-polyhedral oligomeric silsesquioxane organic electronic dye

    Directory of Open Access Journals (Sweden)

    Wangatia Lodrick Makokha

    2015-03-01

    Full Text Available Aggregation-induced quenching is particularly detrimental in perylene diimides, which are characterized by a near-unity fluorescence quantum yield in solution but are far less emissive in the solid state. Previously, perylene diimide has been improved by linking it to the inorganic cage of polyhedral oligomeric silsesquioxanes. As a further study on perylene diimidepolyhedral oligomeric silsesquioxanes, we report on a double functionalized molecular structure, which can be used for substitution at the bay area and as a side group in other materials. Typical solution absorption and emission features of the perylene diimide fragment have been observed in this new reactive perylene diimide-polyhedral oligomeric silsesquioxane. Moreover, reduced stacking during aggregation and spherical particles exhibiting solid fluorescence have been obtained. Organic semiconducting material with enhanced solid state photophysical properties, like solid fluorescence is a subject of great interest owing to its possible high-tech applications in optoelectronic devices.

  18. 28 CFR 540.64 - Press pools.

    Science.gov (United States)

    2010-07-01

    ... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Press pools. 540.64 Section 540.64... PERSONS IN THE COMMUNITY Contact With News Media § 540.64 Press pools. (a) The Warden may establish a press pool whenever he or she determines that the frequency of requests for interviews and...

  19. 7 CFR 1032.7 - Pool plant.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 9 2010-01-01 2009-01-01 true Pool plant. 1032.7 Section 1032.7 Agriculture... Handling Definitions § 1032.7 Pool plant. Pool plant means a plant, unit of plants, or system of plants as specified in paragraphs (a) through (f) of this section, or a plant specified in paragraph (i) of...

  20. 7 CFR 1033.7 - Pool plant.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 9 2010-01-01 2009-01-01 true Pool plant. 1033.7 Section 1033.7 Agriculture... Handling Definitions § 1033.7 Pool plant. Pool plant means a plant, unit of plants, or system of plants as specified in paragraphs (a) through (f) of this section, or a plant specified in paragraph (j) of...

  1. 7 CFR 1124.7 - Pool plant.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 9 2010-01-01 2009-01-01 true Pool plant. 1124.7 Section 1124.7 Agriculture... Regulating Handling Definitions § 1124.7 Pool plant. Pool plant means a plant, unit of plants, or a system of plants as specified in paragraphs (a) through (f) of this section, but excluding a plant specified...

  2. 7 CFR 1126.7 - Pool plant.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 9 2010-01-01 2009-01-01 true Pool plant. 1126.7 Section 1126.7 Agriculture... Handling Definitions § 1126.7 Pool plant. Pool plant means a plant specified in paragraphs (a) through (d) of this section, a unit of plants as specified in paragraph (e) of this section, or a plant...

  3. 7 CFR 1131.7 - Pool plant.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 9 2010-01-01 2009-01-01 true Pool plant. 1131.7 Section 1131.7 Agriculture... Handling Definitions § 1131.7 Pool plant. Pool Plant means a plant or unit of plants specified in paragraphs (a) through (e) of this section, but excluding a plant specified in paragraph (g) of this...

  4. 7 CFR 1007.7 - Pool plant.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 9 2010-01-01 2009-01-01 true Pool plant. 1007.7 Section 1007.7 Agriculture... Handling Definitions § 1007.7 Pool plant. Pool plant means a plant specified in paragraphs (a) through (d) of this section, a unit of plants as specified in paragraph (e) of this section, or a plant...

  5. 7 CFR 1005.7 - Pool plant.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 9 2010-01-01 2009-01-01 true Pool plant. 1005.7 Section 1005.7 Agriculture... Handling Definitions § 1005.7 Pool plant. Pool plant means a plant specified in paragraphs (a) through (d) of this section, a unit of plants as specified in paragraph (e) of this section, or a plant...

  6. 7 CFR 1001.7 - Pool plant.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 9 2010-01-01 2009-01-01 true Pool plant. 1001.7 Section 1001.7 Agriculture... Handling Definitions § 1001.7 Pool plant. Pool plant means a plant, unit of plants, or system of plants as specified in paragraphs (a) through (f) of this section, but excluding a plant described in paragraph (h)...

  7. 7 CFR 1006.7 - Pool plant.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 9 2010-01-01 2009-01-01 true Pool plant. 1006.7 Section 1006.7 Agriculture... Handling Definitions § 1006.7 Pool plant. Pool plant means a plant specified in paragraphs (a) through (d) of this section, a unit of plants as specified in paragraph (e) of this section, or a plant...

  8. 7 CFR 1030.7 - Pool plant.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 9 2010-01-01 2009-01-01 true Pool plant. 1030.7 Section 1030.7 Agriculture... Handling Definitions § 1030.7 Pool plant. Pool plant means a plant, unit of plants, or system of plants as specified in paragraphs (a) through (f) of this section, but excluding a plant specified in paragraph (h)...

  9. p53 oligomerization status modulates cell fate decisions between growth, arrest and apoptosis.

    Science.gov (United States)

    Fischer, Nicholas W; Prodeus, Aaron; Malkin, David; Gariépy, Jean

    2016-12-01

    Mutations in the oligomerization domain of p53 are genetically linked to cancer susceptibility in Li-Fraumeni Syndrome. These mutations typically alter the oligomeric state of p53 and impair its transcriptional activity. Activation of p53 through tetramerization is required for its tumor suppressive function by inducing transcriptional programs that lead to cell fate decisions such as cell cycle arrest or apoptosis. How p53 chooses between these cell fate outcomes remains unclear. Here, we use 5 oligomeric variants of p53, including 2 novel p53 constructs, that yield either monomeric, dimeric or tetrameric forms of p53 and demonstrate that they induce distinct cellular activities and gene expression profiles that lead to different cell fate outcomes. We report that dimeric p53 variants are cytostatic and can arrest cell growth, but lack the ability to trigger apoptosis in p53-null cells. In contrast, p53 tetramers induce rapid apoptosis and cell growth arrest, while a monomeric variant is functionally inactive, supporting cell growth. In particular, the expression of pro-arrest CDKN1A and pro-apoptotic P53AIP1 genes are important cell fate determinants that are differentially regulated by the oligomeric state of p53. This study suggests that the most abundant oligomeric species of p53 present in resting cells, namely p53 dimers, neither promote cell growth or cell death and that shifting the oligomeric state equilibrium of p53 in cells toward monomers or tetramers is a key parameter in p53-based cell fate decisions.

  10. A stable transcription factor complex nucleated by oligomeric AML1–ETO controls leukaemogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Xiao-Jian; Wang, Zhanxin; Wang, Lan; Jiang, Yanwen; Kost, Nils; Soong, T. David; Chen, Wei-Yi; Tang, Zhanyun; Nakadai, Tomoyoshi; Elemento, Olivier; Fischle, Wolfgang; Melnick, Ari; Patel, Dinshaw J.; Nimer, Stephen D.; Roeder, Robert G.

    2013-06-30

    Transcription factors are frequently altered in leukaemia through chromosomal translocation, mutation or aberrant expression. AML1–ETO, a fusion protein generated by the t(8;21) translocation in acute myeloid leukaemia, is a transcription factor implicated in both gene repression and activation. AML1–ETO oligomerization, mediated by the NHR2 domain, is critical for leukaemogenesis, making it important to identify co-regulatory factors that ‘read’ the NHR2 oligomerization and contribute to leukaemogenesis. Here we show that, in human leukaemic cells, AML1–ETO resides in and functions through a stable AML1–ETO-containing transcription factor complex (AETFC) that contains several haematopoietic transcription (co)factors. These AETFC components stabilize the complex through multivalent interactions, provide multiple DNA-binding domains for diverse target genes, co-localize genome wide, cooperatively regulate gene expression, and contribute to leukaemogenesis. Within the AETFC complex, AML1–ETO oligomerization is required for a specific interaction between the oligomerized NHR2 domain and a novel NHR2-binding (N2B) motif in E proteins. Crystallographic analysis of the NHR2–N2B complex reveals a unique interaction pattern in which an N2B peptide makes direct contact with side chains of two NHR2 domains as a dimer, providing a novel model of how dimeric/oligomeric transcription factors create a new protein-binding interface through dimerization/oligomerization. Intriguingly, disruption of this interaction by point mutations abrogates AML1–ETO-induced haematopoietic stem/progenitor cell self-renewal and leukaemogenesis. These results reveal new mechanisms of action of AML1–ETO, and provide a potential therapeutic target in t(8;21)-positive acute myeloid leukaemia.

  11. Requirements and ontology for a G protein-coupled receptor oligomerization knowledge base

    Directory of Open Access Journals (Sweden)

    Parmentier Marc

    2007-05-01

    Full Text Available Abstract Background G Protein-Coupled Receptors (GPCRs are a large and diverse family of membrane proteins whose members participate in the regulation of most cellular and physiological processes and therefore represent key pharmacological targets. Although several bioinformatics resources support research on GPCRs, most of these have been designed based on the traditional assumption that monomeric GPCRs constitute the functional receptor unit. The increase in the frequency and number of reports about GPCR dimerization/oligomerization and the implication of oligomerization in receptor function makes necessary the ability to store and access information about GPCR dimers/oligomers electronically. Results We present here the requirements and ontology (the information scheme to describe oligomers and associated concepts and their relationships for an information system that can manage the elements of information needed to describe comprehensively the phenomena of both homo- and hetero-oligomerization of GPCRs. The comprehensive information management scheme that we plan to use for the development of an intuitive and user-friendly GPCR-Oligomerization Knowledge Base (GPCR-OKB is the result of a community dialog involving experimental and computational colleagues working on GPCRs. Conclusion Our long term goal is to disseminate to the scientific community organized, curated, and detailed information about GPCR dimerization/oligomerization and its related structural context. This information will be reported as close to the data as possible so the user can make his own judgment on the conclusions drawn for a particular study. The requirements and ontology described here will facilitate the development of future information systems for GPCR oligomers that contain both computational and experimental information about GPCR oligomerization. This information is freely accessible at http://www.gpcr-okb.org.

  12. Zn(2+) site engineering at the oligomeric interface of the dopamine transporter

    DEFF Research Database (Denmark)

    Norgaard-Nielsen, Kristine; Norregaard, Lene; Hastrup, Hanne;

    2002-01-01

    of the dopamine transporter (DAT) corresponding to the external end of transmembrane segment 6. Upon binding to this site, which involves a histidine inserted in position 310 (V310H) and the endogenous Cys306 within the same DAT molecule, Zn(2+) potently inhibits [(3)H]dopamine uptake. These data provide indirect...

  13. Hydrogen ions directly regulating the oligomerization state of Photosystem I in intact Spirulina platensis cells

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    H+ concentration induced-monomerization or trimerization of photosystem I (PSI) in cyanobacteria has never been directly observed. In this work, taking characteristic spectra for the trimers and monomers as the indicators, it was experimentally demonstrated that H+ could induce the oligomeric changes of PSI reaction centers in the intact Spirulina ,platensis cells and also in the isolated thylakoid membrane complexes. Especially, the higher concentration of H+ would induce the monomerization while the lower the trimerization, suggesting the electrostatic interaction should be mainly responsible forchanges in the oligomeric state of PSI in Spirulina platensis.

  14. Tip-induced C-H activation and oligomerization of thienoanthracenes.

    Science.gov (United States)

    Dinca, L E; MacLeod, J M; Lipton-Duffin, J; Fu, C; Ma, D; Perepichka, D F; Rosei, F

    2014-08-14

    The tip of a scanning tunneling microscope (STM) can be used to dehydrogenate freely-diffusing tetrathienoanthracene (TTA) molecules on Cu(111), trapping the molecules into metal-coordinated oligomeric structures. The process proceeds at bias voltages above ~3 V and produces organometallic structures identical to those resulting from the thermally-activated cross-coupling of a halogenated analogue. The process appears to be substrate dependent: no oligomerization was observed on Ag(111) or HOPG. This approach demonstrates the possibility of controlled synthesis and nanoscale patterning of 2D oligomer structures on selected surfaces.

  15. POOL development status and production experience

    CERN Document Server

    Chytracek, R; Frank, M; Girone, M; Govi, G; Moscicki, J T; Papadopoulos, I; Schmücker, H; Karr, K; Malon, D; Vaniachine, A; Tanenbaum, W; Xie, Z; Barrass, T; Cioffi, C; IT

    2005-01-01

    The pool of persistent objects for LHC (POOL) project, part of the large Hadron collider (LHC) computing grid (LCG), is now entering its third year of active development. POOL provides the baseline persistency framework for three LHC experiments. It is based on a strict component model, insulating experiment software from a variety of storage technologies. This paper gives a brief overview of the POOL architecture, its main design principles and the experience gained with integration into LHC experiment frameworks. It also presents recent developments in the POOL works areas of relational database abstraction and object storage into relational database management systems (RDBMS) systems.

  16. Patent Pools: Intellectual Property Rights and Competition

    Science.gov (United States)

    Rodriguez, Victor

    2010-01-01

    Patent pools do not correct all problems associated with patent thickets. In this respect, patent pools might not stop the outsider problem from striking pools. Moreover, patent pools can be expensive to negotiate, can exclude patent holders with smaller numbers of patents or enable a group of major players to form a cartel that excludes new competitors. For all the above reasons, patent pools are subject to regulatory clearance because they could result in a monopoly. The aim of this article is to present the relationship between patents and competition in a broad context. PMID:20200607

  17. A NOVEL APPROACH TO SPENT FUEL POOL DECOMMISSIONING

    Energy Technology Data Exchange (ETDEWEB)

    R. L. Demmer

    2011-04-01

    The Idaho National Laboratory (INL) has been at the forefront of developing methods to reduce the cost and schedule of deactivating spent fuel pools (SFP). Several pools have been deactivated at the INL using an underwater approach with divers. These projects provided a basis for the INL cooperation with the Dresden Nuclear Power Station Unit 1 SFP (Exelon Generation Company) deactivation. It represents the first time that a commercial nuclear power plant (NPP) SFP was decommissioned using this underwater coating process. This approach has advantages in many aspects, particularly in reducing airborne contamination and allowing safer, more cost effective deactivation. The INL pioneered underwater coating process was used to decommission three SFPs with a total combined pool volume of over 900,000 gallons. INL provided engineering support and shared project plans to successfully initiate the Dresden project. This report outlines the steps taken by INL and Exelon to decommission SFPs using the underwater coating process. The rationale used to select the underwater coating process and the advantages and disadvantages are described. Special circumstances are also discussed, such as the use of a remotely-operated underwater vehicle to visually and radiologically map the pool areas that were not readily accessible. A larger project, the INTEC-603 SFP in-situ (grouting) deactivation, is reviewed. Several specific areas where special equipment was employed are discussed and a Lessons Learned evaluation is included.

  18. Large-scale pool fires

    Directory of Open Access Journals (Sweden)

    Steinhaus Thomas

    2007-01-01

    Full Text Available A review of research into the burning behavior of large pool fires and fuel spill fires is presented. The features which distinguish such fires from smaller pool fires are mainly associated with the fire dynamics at low source Froude numbers and the radiative interaction with the fire source. In hydrocarbon fires, higher soot levels at increased diameters result in radiation blockage effects around the perimeter of large fire plumes; this yields lower emissive powers and a drastic reduction in the radiative loss fraction; whilst there are simplifying factors with these phenomena, arising from the fact that soot yield can saturate, there are other complications deriving from the intermittency of the behavior, with luminous regions of efficient combustion appearing randomly in the outer surface of the fire according the turbulent fluctuations in the fire plume. Knowledge of the fluid flow instabilities, which lead to the formation of large eddies, is also key to understanding the behavior of large-scale fires. Here modeling tools can be effectively exploited in order to investigate the fluid flow phenomena, including RANS- and LES-based computational fluid dynamics codes. The latter are well-suited to representation of the turbulent motions, but a number of challenges remain with their practical application. Massively-parallel computational resources are likely to be necessary in order to be able to adequately address the complex coupled phenomena to the level of detail that is necessary.

  19. Styrene oligomerization as a molecular probe reaction for Brønsted acidity at the nanoscale

    NARCIS (Netherlands)

    Aramburo Corrales, Luis; Wirick, Sue; Miedema, P.S.; Buurmans, I.L.C.; de Groot, F.M.F.; Weckhuysen, B.M.

    2012-01-01

    The Brønsted acid-catalyzed oligomerization of 4-fluorostyrene has been studied on a series ofH-ZSM-5 zeolite powders, steamed under different conditions, with a combination of UV-Vismicro-spectroscopy and Scanning Transmission X-ray Microscopy (STXM). UV-Vis microspectroscopyand STXM have been used

  20. Selective Antimicrobial Activities and Action Mechanism of Micelles Self-Assembled by Cationic Oligomeric Surfactants.

    Science.gov (United States)

    Zhou, Chengcheng; Wang, Fengyan; Chen, Hui; Li, Meng; Qiao, Fulin; Liu, Zhang; Hou, Yanbo; Wu, Chunxian; Fan, Yaxun; Liu, Libing; Wang, Shu; Wang, Yilin

    2016-02-17

    This work reports that cationic micelles formed by cationic trimeric, tetrameric, and hexameric surfactants bearing amide moieties in spacers can efficiently kill Gram-negative E. coli with a very low minimum inhibitory concentration (1.70-0.93 μM), and do not cause obvious toxicity to mammalian cells at the concentrations used. With the increase of the oligomerization degree, the antibacterial activity of the oligomeric surfactants increases, i.e., hexameric surfactant > tetrameric surfactant > trimeric surfactant. Isothermal titration microcalorimetry, scanning electron microscopy, and zeta potential results reveal that the cationic micelles interact with the cell membrane of E. coli through two processes. First, the integrity of outer membrane of E. coli is disrupted by the electrostatic interaction of the cationic ammonium groups of the surfactants with anionic groups of E. coli, resulting in loss of the barrier function of the outer membrane. The inner membrane then is disintegrated by the hydrophobic interaction of the surfactant hydrocarbon chains with the hydrophobic domains of the inner membrane, leading to the cytoplast leakage. The formation of micelles of these cationic oligomeric surfactants at very low concentration enables more efficient interaction with bacterial cell membrane, which endows the oligomeric surfactants with high antibacterial activity.

  1. Selective lowering of synapsins induced by oligomeric α-synuclein exacerbates memory deficits.

    Science.gov (United States)

    Larson, Megan E; Greimel, Susan J; Amar, Fatou; LaCroix, Michael; Boyle, Gabriel; Sherman, Mathew A; Schley, Hallie; Miel, Camille; Schneider, Julie A; Kayed, Rakez; Benfenati, Fabio; Lee, Michael K; Bennett, David A; Lesné, Sylvain E

    2017-06-06

    Mounting evidence indicates that soluble oligomeric forms of amyloid proteins linked to neurodegenerative disorders, such as amyloid-β (Aβ), tau, or α-synuclein (αSyn) might be the major deleterious species for neuronal function in these diseases. Here, we found an abnormal accumulation of oligomeric αSyn species in AD brains by custom ELISA, size-exclusion chromatography, and nondenaturing/denaturing immunoblotting techniques. Importantly, the abundance of αSyn oligomers in human brain tissue correlated with cognitive impairment and reductions in synapsin expression. By overexpressing WT human αSyn in an AD mouse model, we artificially enhanced αSyn oligomerization. These bigenic mice displayed exacerbated Aβ-induced cognitive deficits and a selective decrease in synapsins. Following isolation of various soluble αSyn assemblies from transgenic mice, we found that in vitro delivery of exogenous oligomeric αSyn but not monomeric αSyn was causing a lowering in synapsin-I/II protein abundance. For a particular αSyn oligomer, these changes were either dependent or independent on endogenous αSyn expression. Finally, at a molecular level, the expression of synapsin genes SYN1 and SYN2 was down-regulated in vivo and in vitro by αSyn oligomers, which decreased two transcription factors, cAMP response element binding and Nurr1, controlling synapsin gene promoter activity. Overall, our results demonstrate that endogenous αSyn oligomers can impair memory by selectively lowering synapsin expression.

  2. Antitumor-promoting activity of oligomeric proanthocyanidins in mouse epidermis in vivo

    Science.gov (United States)

    Mei Xiao Gao; Elisabeth M. Perchellet; Hala U. Gali; Limarie Rodriguez; Richard W. Hemingway; Jean-Pierre Perchellet

    1994-01-01

    The flavanoid catechin and heterogenous samples of oligomeric proanthocyanidins extracted from various sources were compared for their ability to inhibit the biochemical and biological effects of 12-o-tertra-decanoylphorbol-13-acetate (TPA) in mouse epidermis in vivo. Topical applications of catechin fail to alter the hydroperoxide response to TPA but inhibit the...

  3. Monitoring receptor oligomerization by line-scan fluorescence cross-correlation spectroscopy

    NARCIS (Netherlands)

    Hink, M.A.; Postma, M.; Conn, P.M.

    2013-01-01

    Membrane-localized receptor proteins are involved in many signaling cascades, and diffusion and oligomerization are key processes controlling their activity. In order to study these processes in living cells, fluorescence fluctuation spectroscopy techniques have been developed that allow the quantif

  4. Concentration Dependent Ion-Protein Interaction Patterns Underlying Protein Oligomerization Behaviours

    Science.gov (United States)

    Batoulis, Helena; Schmidt, Thomas H.; Weber, Pascal; Schloetel, Jan-Gero; Kandt, Christian; Lang, Thorsten

    2016-04-01

    Salts and proteins comprise two of the basic molecular components of biological materials. Kosmotropic/chaotropic co-solvation and matching ion water affinities explain basic ionic effects on protein aggregation observed in simple solutions. However, it is unclear how these theories apply to proteins in complex biological environments and what the underlying ionic binding patterns are. Using the positive ion Ca2+ and the negatively charged membrane protein SNAP25, we studied ion effects on protein oligomerization in solution, in native membranes and in molecular dynamics (MD) simulations. We find that concentration-dependent ion-induced protein oligomerization is a fundamental chemico-physical principle applying not only to soluble but also to membrane-anchored proteins in their native environment. Oligomerization is driven by the interaction of Ca2+ ions with the carboxylate groups of aspartate and glutamate. From low up to middle concentrations, salt bridges between Ca2+ ions and two or more protein residues lead to increasingly larger oligomers, while at high concentrations oligomers disperse due to overcharging effects. The insights provide a conceptual framework at the interface of physics, chemistry and biology to explain binding of ions to charged protein surfaces on an atomistic scale, as occurring during protein solubilisation, aggregation and oligomerization both in simple solutions and membrane systems.

  5. Styrene oligomerization as a molecular probe reaction for Brønsted acidity at the nanoscale

    NARCIS (Netherlands)

    Aramburo Corrales, Luis; Wirick, Sue; Miedema, P.S.; Buurmans, I.L.C.; de Groot, F.M.F.; Weckhuysen, B.M.

    2012-01-01

    The Brønsted acid-catalyzed oligomerization of 4-fluorostyrene has been studied on a series ofH-ZSM-5 zeolite powders, steamed under different conditions, with a combination of UV-Vismicro-spectroscopy and Scanning Transmission X-ray Microscopy (STXM). UV-Vis microspectroscopyand STXM have been used

  6. Oligomerization of a Glucagon-like Peptide 1 Analog: Bridging Experiment and Simulations

    DEFF Research Database (Denmark)

    Frederiksen, Tine Maja; Sønderby, Pernille; Ryberg, Line A.

    2015-01-01

    The glucagon-like peptide 1 (GLP-1) analog, liraglutide, is a GLP-1 agonist and is used in the treatment of type-2 diabetes mellitus and obesity. From a pharmaceutical perspective, it is important to know the oligomerization state of liraglutide with respect to stability. Compared to GLP-1...

  7. Radioisotope Power System Pool Concept

    Science.gov (United States)

    Rusick, Jeffrey J.; Bolotin, Gary S.

    2015-01-01

    Advanced Radioisotope Power Systems (RPS) for NASA deep space science missions have historically used static thermoelectric-based designs because they are highly reliable, and their radioisotope heat sources can be passively cooled throughout the mission life cycle. Recently, a significant effort to develop a dynamic RPS, the Advanced Stirling Radioisotope Generator (ASRG), was conducted by NASA and the Department of Energy, because Stirling based designs offer energy conversion efficiencies four times higher than heritage thermoelectric designs; and the efficiency would proportionately reduce the amount of radioisotope fuel needed for the same power output. However, the long term reliability of a Stirling based design is a concern compared to thermoelectric designs, because for certain Stirling system architectures the radioisotope heat sources must be actively cooled via the dynamic operation of Stirling converters throughout the mission life cycle. To address this reliability concern, a new dynamic Stirling cycle RPS architecture is proposed called the RPS Pool Concept.

  8. Systematic comparison of the effects of alpha-synuclein mutations on its oligomerization and aggregation.

    Directory of Open Access Journals (Sweden)

    Diana F Lázaro

    2014-11-01

    Full Text Available Aggregation of alpha-synuclein (ASYN in Lewy bodies and Lewy neurites is the typical pathological hallmark of Parkinson's disease (PD and other synucleinopathies. Furthermore, mutations in the gene encoding for ASYN are associated with familial and sporadic forms of PD, suggesting this protein plays a central role in the disease. However, the precise contribution of ASYN to neuronal dysfunction and death is unclear. There is intense debate about the nature of the toxic species of ASYN and little is known about the molecular determinants of oligomerization and aggregation of ASYN in the cell. In order to clarify the effects of different mutations on the propensity of ASYN to oligomerize and aggregate, we assembled a panel of 19 ASYN variants and compared their behaviour. We found that familial mutants linked to PD (A30P, E46K, H50Q, G51D and A53T exhibited identical propensities to oligomerize in living cells, but had distinct abilities to form inclusions. While the A30P mutant reduced the percentage of cells with inclusions, the E46K mutant had the opposite effect. Interestingly, artificial proline mutants designed to interfere with the helical structure of the N-terminal domain, showed increased propensity to form oligomeric species rather than inclusions. Moreover, lysine substitution mutants increased oligomerization and altered the pattern of aggregation. Altogether, our data shed light into the molecular effects of ASYN mutations in a cellular context, and established a common ground for the study of genetic and pharmacological modulators of the aggregation process, opening new perspectives for therapeutic intervention in PD and other synucleinopathies.

  9. Phenolic compounds prevent the oligomerization of α-synuclein and reduce synaptic toxicity.

    Science.gov (United States)

    Takahashi, Ryoichi; Ono, Kenjiro; Takamura, Yusaku; Mizuguchi, Mineyuki; Ikeda, Tokuhei; Nishijo, Hisao; Yamada, Masahito

    2015-09-01

    Lewy bodies, mainly composed of α-synuclein (αS), are pathological hallmarks of Parkinson's disease and dementia with Lewy bodies. Epidemiological studies showed that green tea consumption or habitual intake of phenolic compounds reduced Parkinson's disease risk. We previously reported that phenolic compounds inhibited αS fibrillation and destabilized preformed αS fibrils. Cumulative evidence suggests that low-order αS oligomers are neurotoxic and critical species in the pathogenesis of α-synucleinopathies. To develop disease modifying therapies for α-synucleinopathies, we examined effects of phenolic compounds (myricetin (Myr), curcumin, rosmarinic acid (RA), nordihydroguaiaretic acid, and ferulic acid) on αS oligomerization. Using methods such as photo-induced cross-linking of unmodified proteins, circular dichroism spectroscopy, the electron microscope, and the atomic force microscope, we showed that Myr and RA inhibited αS oligomerization and secondary structure conversion. The nuclear magnetic resonance analysis revealed that Myr directly bound to the N-terminal region of αS, whereas direct binding of RA to monomeric αS was not detected. Electrophysiological assays for long-term potentiation in mouse hippocampal slices revealed that Myr and RA ameliorated αS synaptic toxicity by inhibition of αS oligomerization. These results suggest that Myr and RA prevent the αS aggregation process, reducing the neurotoxicity of αS oligomers. To develop disease modifying therapies for α-synucleinopathies, we examined effects of phenolic compounds on α-synuclein (αS) oligomerization. Phenolic compounds, especially Myricetin (Myr) and Rosmarinic acid (RA), inhibited αS oligomerization and secondary structure conversion. Myr and RA ameliorated αS synaptic toxicity on the experiment of long-term potentiation. Our results suggest that Myr and RA prevent αS aggregation process and reduce the neurotoxicity of αS oligomers. Phenolic compounds are good

  10. Confocal Spectroscopy to Study Dimerization, Oligomerization and Aggregation of Proteins: A Practical Guide.

    Science.gov (United States)

    Gambin, Yann; Polinkovsky, Mark; Francois, Bill; Giles, Nichole; Bhumkar, Akshay; Sierecki, Emma

    2016-04-30

    Protein self-association is a key feature that can modulate the physiological role of proteins or lead to deleterious effects when uncontrolled. Protein oligomerization is a simple way to modify the activity of a protein, as the modulation of binding interfaces allows for self-activation or inhibition, or variation in the selectivity of binding partners. As such, dimerization and higher order oligomerization is a common feature in signaling proteins, for example, and more than 70% of enzymes have the potential to self-associate. On the other hand, protein aggregation can overcome the regulatory mechanisms of the cell and can have disastrous physiological effects. This is the case in a number of neurodegenerative diseases, where proteins, due to mutation or dysregulation later in life, start polymerizing and often fibrillate, leading to the creation of protein inclusion bodies in cells. Dimerization, well-defined oligomerization and random aggregation are often difficult to differentiate and characterize experimentally. Single molecule "counting" methods are particularly well suited to the study of self-oligomerization as they allow observation and quantification of behaviors in heterogeneous conditions. However, the extreme dilution of samples often causes weak complexes to dissociate, and rare events can be overlooked. Here, we discuss a straightforward alternative where the principles of single molecule detection are used at higher protein concentrations to quantify oligomers and aggregates in a background of monomers. We propose a practical guide for the use of confocal spectroscopy to quantify protein oligomerization status and also discuss about its use in monitoring changes in protein aggregation in drug screening assays.

  11. SLC30A3 (ZnT3 oligomerization by dityrosine bonds regulates its subcellular localization and metal transport capacity.

    Directory of Open Access Journals (Sweden)

    Gloria Salazar

    Full Text Available Non-covalent and covalent homo-oligomerization of membrane proteins regulates their subcellular localization and function. Here, we described a novel oligomerization mechanism affecting solute carrier family 30 member 3/zinc transporter 3 (SLC30A3/ZnT3. Oligomerization was mediated by intermolecular covalent dityrosine bonds. Using mutagenized ZnT3 expressed in PC12 cells, we identified two critical tyrosine residues necessary for dityrosine-mediated ZnT3 oligomerization. ZnT3 carrying the Y372F mutation prevented ZnT3 oligomerization, decreased ZnT3 targeting to synaptic-like microvesicles (SLMVs, and decreased resistance to zinc toxicity. Strikingly, ZnT3 harboring the Y357F mutation behaved as a "gain-of-function" mutant as it displayed increased ZnT3 oligomerization, targeting to SLMVs, and increased resistance to zinc toxicity. Single and double tyrosine ZnT3 mutants indicate that the predominant dimeric species is formed between tyrosine 357 and 372. ZnT3 tyrosine dimerization was detected under normal conditions and it was enhanced by oxidative stress. Covalent species were also detected in other SLC30A zinc transporters localized in different subcellular compartments. These results indicate that covalent tyrosine dimerization of a SLC30A family member modulates its subcellular localization and zinc transport capacity. We propose that dityrosine-dependent membrane protein oligomerization may regulate the function of diverse membrane protein in normal and disease states.

  12. A Separate Pool of Cardiac Phospholemman That Does Not Regulate or Associate with the Sodium Pump

    Science.gov (United States)

    Wypijewski, Krzysztof J.; Howie, Jacqueline; Reilly, Louise; Tulloch, Lindsay B.; Aughton, Karen L.; McLatchie, Linda M.; Shattock, Michael J.; Calaghan, Sarah C.; Fuller, William

    2013-01-01

    Phospholemman (PLM), the principal quantitative sarcolemmal substrate for protein kinases A and C in the heart, regulates the cardiac sodium pump. Much like phospholamban, which regulates the related ATPase SERCA, PLM is reported to oligomerize. We investigated subpopulations of PLM in adult rat ventricular myocytes based on phosphorylation status. Co-immunoprecipitation identified two pools of PLM: one not associated with the sodium pump phosphorylated at Ser63 and one associated with the pump, both phosphorylated at Ser68 and unphosphorylated. Phosphorylation of PLM at Ser63 following activation of PKC did not abrogate association of PLM with the pump, so its failure to associate with the pump was not due to phosphorylation at this site. All pools of PLM co-localized to cell surface caveolin-enriched microdomains with sodium pump α subunits, despite the lack of caveolin-binding motif in PLM. Mass spectrometry analysis of phosphospecific immunoprecipitation reactions revealed no unique protein interactions for Ser63-phosphorylated PLM, and cross-linking reagents also failed to identify any partner proteins for this pool. In lysates from hearts of heterozygous transgenic animals expressing wild type and unphosphorylatable PLM, Ser63-phosphorylated PLM co-immunoprecipitated unphosphorylatable PLM, confirming the existence of PLM multimers. Dephosphorylation of the PLM multimer does not change sodium pump activity. Hence like phospholamban, PLM exists as a pump-inhibiting monomer and an unassociated oligomer. The distribution of different PLM phosphorylation states to different pools may be explained by their differential proximity to protein phosphatases rather than a direct effect of phosphorylation on PLM association with the pump. PMID:23532852

  13. BAX insertion, oligomerization, and outer membrane permeabilization in brain mitochondria: role of permeability transition and SH-redox regulation

    Science.gov (United States)

    Brustovetsky, Tatiana; Li, Tsyregma; Yang, Youyun; Zhang, Jiang-Ting; Antonsson, Bruno; Brustovetsky, Nickolay

    2010-01-01

    BAX cooperates with truncated BID (tBID) and Ca2+ in permeabilizing the outer mitochondrial membrane (OMM) and releasing mitochondrial apoptogenic proteins. The mechanisms of this cooperation are still unclear. Here we show that in isolated brain mitochondria, recombinant BAX readily self-integrates/oligomerizes in the OMM but produces only a minuscule release of cytochrome c, indicating that BAX insertion/oligomerization in the OMM does not always lead to massive OMM permeabilization. Ca2+ in a mitochondrial permeability transition (mPT)-dependent and recombinant tBID in an mPT-independent manner promoted BAX insertion/oligomerization in the OMM and augmented cytochrome c release. Neither tBID nor Ca2+ induced BAX oligomerization in the solution without mitochondria, suggesting that BAX oligomerization required interaction with the organelles and followed rather than preceded BAX insertion in the OMM. Recombinant Bcl-xL failed to prevent BAX insertion/oligomerization in the OMM but strongly attenuated cytochrome c release. On the other hand, a reducing agent, dithiothreitol (DTT), inhibited BAX insertion/oligomerization augmented by tBID or Ca2+ and suppressed the BAX-mediated release of cytochrome c and Smac/DIABLO but failed to inhibit Ca2+-induced swelling. Altogether, these data suggest that in brain mitochondria, BAX insertion/oligomerization can be dissociated from OMM permeabilization and that tBID and Ca2+ stimulate BAX insertion/oligomerization and BAX-mediated OMM permeabilization by different mechanisms involving mPT induction and modulation of the SH-redox state. PMID:20655869

  14. Synthesis of Nonionic Oligomeric Manganese(Ⅱ) Complexes and Investigation of Their Toxicity and Τ1-Relaxation Enhancement

    Institute of Scientific and Technical Information of China (English)

    YU Kai-chao; LI Ying-xia; ZHOU Jin-lan; DING Shang-wu; YE Chao-hui

    2005-01-01

    Six nonionic oligomeric manganese(Ⅱ) complexes with oligomeric phosphate-polyglycol-EDTA ester ligands were synthesized and characterized. The longitudinal relaxivities of these complexes were measured. One of these complexes, which showed the highest relaxivity, was chosen to be used in the acute toxicity test and the Τ1-weighted imaging experiment. It has been found that compared to Gd-DTPA, this nonionic oligomeric Mn(Ⅱ) complex exhibits no acute toxicity, generates highly enhanced MRI signals and increases the intention time in the rat liver tissue.

  15. DNA structure modulates the oligomerization properties of the AAV initiator protein Rep68.

    Directory of Open Access Journals (Sweden)

    Jorge Mansilla-Soto

    2009-07-01

    Full Text Available Rep68 is a multifunctional protein of the adeno-associated virus (AAV, a parvovirus that is mostly known for its promise as a gene therapy vector. In addition to its role as initiator in viral DNA replication, Rep68 is essential for site-specific integration of the AAV genome into human chromosome 19. Rep68 is a member of the superfamily 3 (SF3 helicases, along with the well-studied initiator proteins simian virus 40 large T antigen (SV40-LTag and bovine papillomavirus (BPV E1. Structurally, SF3 helicases share two domains, a DNA origin interaction domain (OID and an AAA(+ motor domain. The AAA(+ motor domain is also a structural feature of cellular initiators and it functions as a platform for initiator oligomerization. Here, we studied Rep68 oligomerization in vitro in the presence of different DNA substrates using a variety of biophysical techniques and cryo-EM. We found that a dsDNA region of the AAV origin promotes the formation of a complex containing five Rep68 subunits. Interestingly, non-specific ssDNA promotes the formation of a double-ring Rep68, a known structure formed by the LTag and E1 initiator proteins. The Rep68 ring symmetry is 8-fold, thus differing from the hexameric rings formed by the other SF3 helicases. However, similiar to LTag and E1, Rep68 rings are oriented head-to-head, suggesting that DNA unwinding by the complex proceeds bidirectionally. This novel Rep68 quaternary structure requires both the DNA binding and AAA(+ domains, indicating cooperativity between these regions during oligomerization in vitro. Our study clearly demonstrates that Rep68 can oligomerize through two distinct oligomerization pathways, which depend on both the DNA structure and cooperativity of Rep68 domains. These findings provide insight into the dynamics and oligomeric adaptability of Rep68 and serve as a step towards understanding the role of this multifunctional protein during AAV DNA replication and site-specific integration.

  16. GIPP: Geophysical Instrument Pool Potsdam

    Directory of Open Access Journals (Sweden)

    Christian Haberland

    2016-04-01

    Full Text Available The Geophysical Instrument Pool Potsdam (GIPP consists of field instruments, sensors and equipment for temporary seismological studies (both controlled source and earthquake seismology as well as for magnetotelluric (electromagnetic experiments. These instruments are mainly mobile digital recorders, broadband seis­mometers and short period sensors, and they are used to reveal the subsurface structure and to investigate earth­quakes. Sensors for magnetotellurics include induction coil and fluxgate magnetometers and non-polarizing silver / silver-chloride electrodes. It is operated by the Helmholtz Centre Potsdam GFZ German Research Centre for Geosciences. The instru­ment facility is open to all academic applicants, both national and international. Instrument applications are evalu­ated and ranked by an external steering board. Currently, for seismological applications >850 geophysical recorders, >170 broadband seis­mo­meters and >1300 short period geophones are available (among others. Available for magnetotelluric experiments are > 50 real-time data-loggers, >150 induction coils, and >500 electrodes. User guidelines and data policy are in force and data archives are provided (standard exchange formats.

  17. Sulfur cycling in two Dutch moorland pools

    NARCIS (Netherlands)

    Marnette, E.C.L.

    1993-01-01

    Due to atmospheric acid deposition, the chemistry of many moorland pools has changed, resulting in changes in their fauna and flora. Most moorland pools are sensitive to acid loading because underlying and surrounding soils are low in chemical buffering capacity. Biological processes in the

  18. 10 CFR 36.33 - Irradiator pools.

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 1 2010-01-01 2010-01-01 false Irradiator pools. 36.33 Section 36.33 Energy NUCLEAR REGULATORY COMMISSION LICENSES AND RADIATION SAFETY REQUIREMENTS FOR IRRADIATORS Design and Performance... indicator must be provided in a clearly visible location to indicate if the pool water level is below the...

  19. Sulfur cycling in two Dutch moorland pools.

    NARCIS (Netherlands)

    Marnette, E.C.L.

    1993-01-01

    Due to atmospheric acid deposition, the chemistry of many moorland pools has changed, resulting in changes in their fauna and flora. Most moorland pools are sensitive to acid loading because underlying and surrounding soils are low in chemical buffering capacity. Biological processes in the sediment

  20. Screening of Drugs Inhibiting In vitro Oligomerization of Cu/Zn-Superoxide Dismutase with a Mutation Causing Amyotrophic Lateral Sclerosis.

    Science.gov (United States)

    Anzai, Itsuki; Toichi, Keisuke; Tokuda, Eiichi; Mukaiyama, Atsushi; Akiyama, Shuji; Furukawa, Yoshiaki

    2016-01-01

    Dominant mutations in Cu/Zn-superoxide dismutase (SOD1) gene have been shown to cause a familial form of amyotrophic lateral sclerosis (SOD1-ALS). A major pathological hallmark of this disease is abnormal accumulation of mutant SOD1 oligomers in the affected spinal motor neurons. While no effective therapeutics for SOD1-ALS is currently available, SOD1 oligomerization will be a good target for developing cures of this disease. Recently, we have reproduced the formation of SOD1 oligomers abnormally cross-linked via disulfide bonds in a test tube. Using our in vitro model of SOD1 oligomerization, therefore, we screened 640 FDA-approved drugs for inhibiting the oligomerization of SOD1 proteins, and three effective classes of chemical compounds were identified. Those hit compounds will provide valuable information on the chemical structures for developing a novel drug candidate suppressing the abnormal oligomerization of mutant SOD1 and possibly curing the disease.

  1. Pool scrubbing models for iodine components

    Energy Technology Data Exchange (ETDEWEB)

    Fischer, K. [Battelle Ingenieurtechnik GmbH, Eschborn (Germany)

    1996-12-01

    Pool scrubbing is an important mechanism to retain radioactive fission products from being carried into the containment atmosphere or into the secondary piping system. A number of models and computer codes has been developed to predict the retention of aerosols and fission product vapours that are released from the core and injected into water pools of BWR and PWR type reactors during severe accidents. Important codes in this field are BUSCA, SPARC and SUPRA. The present paper summarizes the models for scrubbing of gaseous Iodine components in these codes, discusses the experimental validation, and gives an assessment of the state of knowledge reached and the open questions which persist. The retention of gaseous Iodine components is modelled by the various codes in a very heterogeneous manner. Differences show up in the chemical species considered, the treatment of mass transfer boundary layers on the gaseous and liquid sides, the gas-liquid interface geometry, calculation of equilibrium concentrations and numerical procedures. Especially important is the determination of the pool water pH value. This value is affected by basic aerosols deposited in the water, e.g. Cesium and Rubidium compounds. A consistent model requires a mass balance of these compounds in the pool, thus effectively coupling the pool scrubbing phenomena of aerosols and gaseous Iodine species. Since the water pool conditions are also affected by drainage flow of condensate water from different regions in the containment, and desorption of dissolved gases on the pool surface is determined by the gas concentrations above the pool, some basic limitations of specialized pool scrubbing codes are given. The paper draws conclusions about the necessity of coupling between containment thermal-hydraulics and pool scrubbing models, and proposes ways of further simulation model development in order to improve source term predictions. (author) 2 tabs., refs.

  2. Near-Field Optical Microscopy of Defects in Cholesteric Oligomeric Liquid Crystal Films

    Energy Technology Data Exchange (ETDEWEB)

    Lukishova, S.G.; Schmid, A.W.

    2006-08-18

    This paper describes formation of 2-D hexagonal structures with a periodicity ~0.5-0.8 um in the defects of thin films of cholesteric oligomeric liquid crystals prepared by the evaporation of the solvent from the oligomer solution on the substrate. These regular arrays were observed by scanning near-field optical and concurrent atomic force microscopy. The mechanisms considered are both Benard-Marangoni and buoyancy conventions induced by solvent evaporation and air-bubble creation around the condensed water droplets from the air during evaporative cooling. Hexagonal structures prepared by this method can be used in photonic devices for emission enhancement, for instance, in liquid crystal lasers and single photon sources with oligomeric liquid crystal hosts.

  3. Structural polymorphism in the N-terminal oligomerization domain of NPM1.

    Science.gov (United States)

    Mitrea, Diana M; Grace, Christy R; Buljan, Marija; Yun, Mi-Kyung; Pytel, Nicholas J; Satumba, John; Nourse, Amanda; Park, Cheon-Gil; Madan Babu, M; White, Stephen W; Kriwacki, Richard W

    2014-03-25

    Nucleophosmin (NPM1) is a multifunctional phospho-protein with critical roles in ribosome biogenesis, tumor suppression, and nucleolar stress response. Here we show that the N-terminal oligomerization domain of NPM1 (Npm-N) exhibits structural polymorphism by populating conformational states ranging from a highly ordered, folded pentamer to a highly disordered monomer. The monomer-pentamer equilibrium is modulated by posttranslational modification and protein binding. Phosphorylation drives the equilibrium in favor of monomeric forms, and this effect can be reversed by Npm-N binding to its interaction partners. We have identified a short, arginine-rich linear motif in NPM1 binding partners that mediates Npm-N oligomerization. We propose that the diverse functional repertoire associated with NPM1 is controlled through a regulated unfolding mechanism signaled through posttranslational modifications and intermolecular interactions.

  4. Domain architecture and oligomerization properties of the paramyxovirus PIV 5 hemagglutinin-neuraminidase (HN) protein.

    Science.gov (United States)

    Yuan, Ping; Leser, George P; Demeler, Borries; Lamb, Robert A; Jardetzky, Theodore S

    2008-09-01

    The mechanism by which the paramyxovirus hemagglutinin-neuraminidase (HN) protein couples receptor binding to activation of virus entry remains to be fully understood, but the HN stalk is thought to play an important role in the process. We have characterized ectodomain constructs of the parainfluenza virus 5 HN to understand better the underlying architecture and oligomerization properties that may influence HN functions. The PIV 5 neuraminidase (NA) domain is monomeric whereas the ectodomain forms a well-defined tetramer. The HN stalk also forms tetramers and higher order oligomers with high alpha-helical content. Together, the data indicate that the globular NA domains form weak intersubunit interactions at the end of the HN stalk tetramer, while stabilizing the stalk and overall oligomeric state of the ectodomain. Electron microscopy of the HN ectodomain reveals flexible arrangements of the NA and stalk domains, which may be important for understanding how these two HN domains impact virus entry.

  5. Crystal structure of listeriolysin O reveals molecular details of oligomerization and pore formation

    Science.gov (United States)

    Köster, Stefan; van Pee, Katharina; Hudel, Martina; Leustik, Martin; Rhinow, Daniel; Kühlbrandt, Werner; Chakraborty, Trinad; Yildiz, Özkan

    2014-04-01

    Listeriolysin O (LLO) is an essential virulence factor of Listeria monocytogenes that causes listeriosis. Listeria monocytogenes owes its ability to live within cells to the pH- and temperature-dependent pore-forming activity of LLO, which is unique among cholesterol-dependent cytolysins. LLO enables the bacteria to cross the phagosomal membrane and is also involved in activation of cellular processes, including the modulation of gene expression or intracellular Ca2+ oscillations. Neither the pore-forming mechanism nor the mechanisms triggering the signalling processes in the host cell are known in detail. Here, we report the crystal structure of LLO, in which we identified regions important for oligomerization and pore formation. Mutants were characterized by determining their haemolytic and Ca2+ uptake activity. We analysed the pore formation of LLO and its variants on erythrocyte ghosts by electron microscopy and show that pore formation requires precise interface interactions during toxin oligomerization on the membrane.

  6. Effect of acid density of HZSM-5 on the oligomerization of ethylene in FCC dry gas

    Institute of Scientific and Technical Information of China (English)

    Xue Ding; Shan Geng; Chunyi Li; Chaohe Yang; Guohui Wang

    2009-01-01

    The oligomerization of ethylene in FCC dry gas over HZSM-5 catalyst with different Si/Al2 ratios was studied. The effect of acid density of catalyst on the oligomerization of ethylene was discussed. By increasing the acid density of catalyst, ethylene conversion showed a linear increase, while the yields of olefins decreased when the acid density of catalyst exceeded 0.14 mmolNH3/g owing to a promotion of hydrogen transfer reaction. Through comparing the average distance between acid sites on catalyst with kinetic diameters of olefins, it was found that the dimerization of ethylene was not restrained by the sparse distribution of acid sites, while the hydrogen transfer reaction of C3 and C4 olefins was limited. On these bases, a conclusion is proposed that the dimerization of ethylene proceeded via Eley-Rideal mechanism, while the hydrogen transfer reaction of C3 and C4 olefins followed the Langmuir-Hinshelwood mechanism.

  7. Molecular basis for oligomeric-DNA binding and episome maintenance by KSHV LANA.

    Directory of Open Access Journals (Sweden)

    John F Domsic

    Full Text Available LANA is the KSHV-encoded terminal repeat binding protein essential for viral replication and episome maintenance during latency. We have determined the X-ray crystal structure of LANA C-terminal DNA binding domain (LANADBD to reveal its capacity to form a decameric ring with an exterior DNA binding surface. The dimeric core is structurally similar to EBV EBNA1 with an N-terminal arm that regulates DNA binding and is required for replication function. The oligomeric interface between LANA dimers is dispensable for single site DNA binding, but is required for cooperative DNA binding, replication function, and episome maintenance. We also identify a basic patch opposite of the DNA binding surface that is responsible for the interaction with BRD proteins and contributes to episome maintenance function. The structural features of LANADBD suggest a novel mechanism of episome maintenance through DNA-binding induced oligomeric assembly.

  8. Multigene expression in stable CHO cell pools generated with the piggyBac transposon system.

    Science.gov (United States)

    Balasubramanian, Sowmya; Wurm, Florian M; Hacker, David L

    2016-09-01

    Heterogenous populations of recombinant cells (cell pools) stably expressing 1-4 transgenes were generated from Chinese hamster overy (CHO) cells with the piggyBac (PB) transposon system. The cell pools produced different combinations of three model proteins-enhanced green fluorescent protein (EGFP), secreted alkaline phosphatase (SEAP), and a monoclonal IgG1 antibody. Each transgene was present on a separate PB donor plasmid with either the same or a different selection gene. In both cases, we obtained PB-derived cell pools with higher recombinant protein yields than from cell pools generated by conventional gene delivery. In PB-derived cell pools generated using a single selection agent, both protein production and the number of integrated copies of each transgene declined as the number of transfected transgenes increased. However, the total number of integrated transgenes was similar regardless of the number of different transgenes transfected. For PB-derived cell pools generated by selection of each transgene with a different selection agent, the total number of integrated transgenes increased with the number of transfected transgenes. The results suggest that the generation of cell pools producing multiple recombinant proteins is feasible and that the method is more efficient when each individual transgene is selected with a different marker. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1308-1317, 2016.

  9. Three-Dimensional Models of the Oligomeric Human Asialoglycoprotein Receptor (ASGP-R)

    OpenAIRE

    Anna Maria Bianucci; Ilaria Massarelli; Federica Chiellini; Emo Chiellini

    2010-01-01

    The work presented here is aimed at suggesting plausible hypotheses for functional oligomeric forms of the human asialoglycoprotein receptor (ASGP-R), by applying a combination of different computational techniques. The functional ASGP-R is a hetero-oligomer, that comprises of several subunits of two different kinds (H1 and H2), which are highly homologous. Its stoichiometry is still unknown. An articulated step-wise modeling protocol was used in order to build the receptor model in a minimal...

  10. Alternative Models of Iron and Cobalt Catalysts for Ethylene Oligomerization and Polymerization

    Institute of Scientific and Technical Information of China (English)

    Katrin; Wedeking; Sherrif; Adewuyi; Maliha; Asma; Igor; Vystorop; Saliu; Amolegbe; Elena; Novikova

    2007-01-01

    1 Results Great progresses have been made in the field of transition metal-based complexes as catalytic precursors for olefin oligomerization and polymerization,in which the core subjects will remain as "know and how" to develop novel catalysts both in academic and industrial consideration.The key advantage of iron and cobalt catalyst for ethylene polymerization is to produce vinyl-type polyethylenes.Therefore following the pioneering works of bis(imino) pyridyl iron and cobalt catalyst by Brookhart[1] ...

  11. Oligomerization of DH Domain Is Essential for Dbl-Induced Transformation

    OpenAIRE

    Zhu, Kejin; Debreceni, Balazs; Bi, Feng; Zheng, Yi

    2001-01-01

    The dbl oncogene product (onco-Dbl) is the prototype member of a family of guanine nucleotide exchange factors (GEFs) for Rho GTPases. The Dbl homology (DH) domain of onco-Dbl is responsible for the GEF catalytic activity, and the DH domain, together with the immediately adjacent pleckstrin homology (PH) domain, constitutes the minimum module bearing transforming function. In the present study, we demonstrate that the onco-Dbl protein exists in oligomeric form in vitro and in cells. The oligo...

  12. Membrane Permeabilization by Oligomeric α-Synuclein: In Search of the Mechanism.

    Directory of Open Access Journals (Sweden)

    Bart D van Rooijen

    Full Text Available BACKGROUND: The question of how the aggregation of the neuronal protein α-synuclein contributes to neuronal toxicity in Parkinson's disease has been the subject of intensive research over the past decade. Recently, attention has shifted from the amyloid fibrils to soluble oligomeric intermediates in the α-synuclein aggregation process. These oligomers are hypothesized to be cytotoxic and to permeabilize cellular membranes, possibly by forming pore-like complexes in the bilayer. Although the subject of α-synuclein oligomer-membrane interactions has attracted much attention, there is only limited evidence that supports the pore formation by α-synuclein oligomers. In addition the existing data are contradictory. METHODOLOGY/PRINCIPAL FINDINGS: Here we have studied the mechanism of lipid bilayer disruption by a well-characterized α-synuclein oligomer species in detail using a number of in vitro bilayer systems and assays. Dye efflux from vesicles induced by oligomeric α-synuclein was found to be a fast all-or-none process. Individual vesicles swiftly lose their contents but overall vesicle morphology remains unaltered. A newly developed assay based on a dextran-coupled dye showed that non-equilibrium processes dominate the disruption of the vesicles. The membrane is highly permeable to solute influx directly after oligomer addition, after which membrane integrity is partly restored. The permeabilization of the membrane is possibly related to the intrinsic instability of the bilayer. Vesicles composed of negatively charged lipids, which are generally used for measuring α-synuclein-lipid interactions, were unstable to protein adsorption in general. CONCLUSIONS/SIGNIFICANCE: The dye efflux from negatively charged vesicles upon addition of α-synuclein has been hypothesized to occur through the formation of oligomeric membrane pores. However, our results show that the dye efflux characteristics are consistent with bilayer defects caused by

  13. Oligomeric state regulated trafficking of human platelet-activating factor acetylhydrolase type-II.

    Science.gov (United States)

    Monillas, Elizabeth S; Caplan, Jeffrey L; Thévenin, Anastasia F; Bahnson, Brian J

    2015-05-01

    The intracellular enzyme platelet-activating factor acetylhydrolase type-II (PAFAH-II) hydrolyzes platelet-activating factor and oxidatively fragmented phospholipids. PAFAH-II in its resting state is mainly cytoplasmic, and it responds to oxidative stress by becoming increasingly bound to endoplasmic reticulum and Golgi membranes. Numerous studies have indicated that this enzyme is essential for protecting cells from oxidative stress induced apoptosis. However, the regulatory mechanism of the oxidative stress response by PAFAH-II has not been fully resolved. Here, changes to the oligomeric state of human PAFAH-II were investigated as a potential regulatory mechanism toward enzyme trafficking. Native PAGE analysis in vitro and photon counting histogram within live cells showed that PAFAH-II is both monomeric and dimeric. A Gly-2-Ala site-directed mutation of PAFAH-II demonstrated that the N-terminal myristoyl group is required for homodimerization. Additionally, the distribution of oligomeric PAFAH-II is distinct within the cell; homodimers of PAFAH-II were localized to the cytoplasm while monomers were associated to the membranes of the endoplasmic reticulum and Golgi. We propose that the oligomeric state of PAFAH-II drives functional protein trafficking. PAFAH-II localization to the membrane is critical for substrate acquisition and effective oxidative stress protection. It is hypothesized that the balance between monomer and dimer serves as a regulatory mechanism of a PAFAH-II oxidative stress response.

  14. Recursive directional ligation by plasmid reconstruction allows rapid and seamless cloning of oligomeric genes.

    Science.gov (United States)

    McDaniel, Jonathan R; Mackay, J Andrew; Quiroz, Felipe García; Chilkoti, Ashutosh

    2010-04-12

    This paper reports a new strategy, recursive directional ligation by plasmid reconstruction (PRe-RDL), to rapidly clone highly repetitive polypeptides of any sequence and specified length over a large range of molecular weights. In a single cycle of PRe-RDL, two halves of a parent plasmid, each containing a copy of an oligomer, are ligated together, thereby dimerizing the oligomer and reconstituting a functional plasmid. This process is carried out recursively to assemble an oligomeric gene with the desired number of repeats. PRe-RDL has several unique features that stem from the use of type IIs restriction endonucleases: first, PRe-RDL is a seamless cloning method that leaves no extraneous nucleotides at the ligation junction. Because it uses type IIs endonucleases to ligate the two halves of the plasmid, PRe-RDL also addresses the major limitation of RDL in that it abolishes any restriction on the gene sequence that can be oligomerized. The reconstitution of a functional plasmid only upon successful ligation in PRe-RDL also addresses two other limitations of RDL: the significant background from self-ligation of the vector observed in RDL, and the decreased efficiency of ligation due to nonproductive circularization of the insert. PRe-RDL can also be used to assemble genes that encode different sequences in a predetermined order to encode block copolymers or append leader and trailer peptide sequences to the oligomerized gene.

  15. Rapid synthesis of cyclic oligomeric depsipeptides with positional, stereochemical, and macrocycle size distribution control.

    Science.gov (United States)

    Batiste, Suzanne M; Johnston, Jeffrey N

    2016-12-27

    Macrocyclic small molecules are attractive tools in the development of sensors, new materials, and therapeutics. Within early-stage drug discovery, they are increasingly sought for their potential to interact with broad surfaces of peptidic receptors rather than within their narrow folds and pockets. Cyclization of linear small molecule precursors is a straightforward strategy to constrain conformationally mobile motifs, but forging a macrocycle bond typically becomes more difficult at larger ring sizes. We report the development of a general approach to discrete collections of oligomeric macrocyclic depsipeptides using an oligomerization/macrocyclization process governed by a series of Mitsunobu reactions of hydroxy acid monomers. Ring sizes of 18, 24, 30, and 36 are formed in a single reaction from a didepsipeptide, whereas sizes of 24, 36, and 60 result from a tetradepsipeptide. The ring-size selectivity inherent to the approach can be modulated by salt additives that enhance the formation of specific ring sizes. Use of chemical synthesis to prepare the monomers suggests broad access to functionally and stereochemically diverse collections of natural product-like oligodepsipeptide macrocycles. Two cyclodepsipeptide natural products were prepared along with numerous unnatural oligomeric congeners to provide rapid access to discrete collections of complex macrocyclic small molecules from medium (18) to large (60) ring sizes.

  16. Phosphomimetic mutations enhance oligomerization of phospholemman and modulate its interaction with the Na/K-ATPase.

    Science.gov (United States)

    Song, Qiujing; Pallikkuth, Sandeep; Bossuyt, Julie; Bers, Donald M; Robia, Seth L

    2011-03-18

    Na/K-ATPase (NKA) activity is dynamically regulated by an inhibitory interaction with a small transmembrane protein, phospholemman (PLM). Inhibition is relieved upon PLM phosphorylation. Phosphorylation may alter how PLM interacts with NKA and/or itself, but details of these interactions are unknown. To address this, we quantified FRET between PLM and its regulatory target NKA in live cells. Phosphorylation of PLM was mimicked by mutation S63E (PKC site), S68E (PKA/PKC site), or S63E/S68E. The dependence of FRET on protein expression in live cells yielded information about the structure and binding affinity of the PLM-NKA regulatory complex. PLM phosphomimetic mutations altered the quaternary structure of the regulatory complex and reduced the apparent affinity of the PLM-NKA interaction. The latter effect was likely due to increased oligomerization of PLM phosphomimetic mutants, as suggested by PLM-PLM FRET measurements. Distance constraints obtained by FRET suggest that phosphomimetic mutations slightly alter the oligomer quaternary conformation. Photon-counting histogram measurements revealed that the major PLM oligomeric species is a tetramer. We conclude that phosphorylation of PLM increases its oligomerization into tetramers, decreases its binding to NKA, and alters the structures of both the tetramer and NKA regulatory complex.

  17. Structural basis for the oligomerization of the MADS domain transcription factor SEPALLATA3 in Arabidopsis.

    Science.gov (United States)

    Puranik, Sriharsha; Acajjaoui, Samira; Conn, Simon; Costa, Luca; Conn, Vanessa; Vial, Anthony; Marcellin, Romain; Melzer, Rainer; Brown, Elizabeth; Hart, Darren; Theißen, Günter; Silva, Catarina S; Parcy, François; Dumas, Renaud; Nanao, Max; Zubieta, Chloe

    2014-09-01

    In plants, MADS domain transcription factors act as central regulators of diverse developmental pathways. In Arabidopsis thaliana, one of the most central members of this family is SEPALLATA3 (SEP3), which is involved in many aspects of plant reproduction, including floral meristem and floral organ development. SEP3 has been shown to form homo and heterooligomeric complexes with other MADS domain transcription factors through its intervening (I) and keratin-like (K) domains. SEP3 function depends on its ability to form specific protein-protein complexes; however, the atomic level determinants of oligomerization are poorly understood. Here, we report the 2.5-Å crystal structure of a small portion of the intervening and the complete keratin-like domain of SEP3. The domains form two amphipathic alpha helices separated by a rigid kink, which prevents intramolecular association and presents separate dimerization and tetramerization interfaces comprising predominantly hydrophobic patches. Mutations to the tetramerization interface demonstrate the importance of highly conserved hydrophobic residues for tetramer stability. Atomic force microscopy was used to show SEP3-DNA interactions and the role of oligomerization in DNA binding and conformation. Based on these data, the oligomerization patterns of the larger family of MADS domain transcription factors can be predicted and manipulated based on the primary sequence. © 2014 American Society of Plant Biologists. All rights reserved.

  18. Functional foods effective for hepatitis C: Identification of oligomeric proanthocyanidin and its action mechanism

    Institute of Scientific and Technical Information of China (English)

    Yo-ichi; Ishida; Masahiko; Takeshita; Hiroaki; Kataoka

    2014-01-01

    Hepatitis C virus(HCV)is a major cause of viral hepatitis and currently infects approximately 170 million people worldwide.An infection by HCV causes high rates of chronic hepatitis(>75%)and progresses to liver cirrhosis and hepatocellular carcinoma ultimately.HCV can be eliminated by a combination of pegylatedα-interferon and the broad-spectrum antiviral drug ribavirin;however,this treatment is still associated with poor efficacy and tolerability and is often accompanied by serious side-effects.While some novel direct-actingantivirals against HCV have been developed recently,high medical costs limit the access to the therapy in cost-sensitive countries.To search for new natural anti-HCV agents,we screened local agricultural products for their suppressive activities against HCV replication using the HCV replicon cell system in vitro.We found a potent inhibitor of HCV RNA expression in the extracts of blueberry leaves and then identified oligomeric proanthocyanidin as the active ingredient.Further investigations into the action mechanism of oligomeric proanthocyanidin suggested that it is an inhibitor of heterogeneous nuclear ribonucleoproteins(hn RNPs)such as hn RNP A2/B1.In this review,we presented an overview of functional foods and ingredients efficient for HCV infection,the chemical structural characteristics of oligomeric proanthocyanidin,and its action mechanism.

  19. Physiological relevance of plant 2-Cys peroxiredoxin overoxidation level and oligomerization status.

    Science.gov (United States)

    Cerveau, Delphine; Ouahrani, Djelloul; Marok, Mohamed Amine; Blanchard, Laurence; Rey, Pascal

    2016-01-01

    Peroxiredoxins are ubiquitous thioredoxin-dependent peroxidases presumed to display, upon environmental constraints, a chaperone function resulting from a redox-dependent conformational switch. In this work, using biochemical and genetic approaches, we aimed to unravel the factors regulating the redox status and the conformation of the plastidial 2-Cys peroxiredoxin (2-Cys PRX) in plants. In Arabidopsis, we show that in optimal growth conditions, the overoxidation level mainly depends on the availability of thioredoxin-related electron donors, but not on sulfiredoxin, the enzyme reducing the 2-Cys PRX overoxidized form. We also observed that upon various physiological temperature, osmotic and light stress conditions, the overoxidation level and oligomerization status of 2-Cys PRX can moderately vary depending on the constraint type. Further, no major change was noticed regarding protein conformation in water-stressed Arabidopsis, barley and potato plants, whereas species-dependent up- and down-variations in overoxidation were observed. In contrast, both 2-Cys PRX overoxidation and oligomerization were strongly induced during a severe oxidative stress generated by methyl viologen. From these data, revealing that the oligomerization status of plant 2-Cys PRX does not exhibit important variation and is not tightly linked to the protein redox status upon physiologically relevant environmental constraints, the possible in planta functions of 2-Cys PRX are discussed.

  20. Structural reorganisation and potential toxicity of oligomeric species formed during the assembly of amyloid fibrils.

    Directory of Open Access Journals (Sweden)

    Mookyung Cheon

    2007-09-01

    Full Text Available Increasing evidence indicates that oligomeric protein assemblies may represent the molecular species responsible for cytotoxicity in a range of neurological disorders including Alzheimer and Parkinson diseases. We use all-atom computer simulations to reveal that the process of oligomerization can be divided into two steps. The first is characterised by a hydrophobic coalescence resulting in the formation of molten oligomers in which hydrophobic residues are sequestered away from the solvent. In the second step, the oligomers undergo a process of reorganisation driven by interchain hydrogen bonding interactions that induce the formation of beta sheet rich assemblies in which hydrophobic groups can become exposed. Our results show that the process of aggregation into either ordered or amorphous species is largely determined by a competition between the hydrophobicity of the amino acid sequence and the tendency of polypeptide chains to form arrays of hydrogen bonds. We discuss how the increase in solvent-exposed hydrophobic surface resulting from such a competition offers an explanation for recent observations concerning the cytotoxicity of oligomeric species formed prior to mature amyloid fibrils.

  1. Non-Enzymatic Oligomerization of 3’, 5’ Cyclic AMP

    Science.gov (United States)

    Costanzo, Giovanna; Pino, Samanta; Timperio, Anna Maria; Šponer, Judit E.; Šponer, Jiří; Nováková, Olga; Šedo, Ondrej; Zdráhal, Zbyněk; Di Mauro, Ernesto

    2016-01-01

    Recent studies illustrate that short oligonucleotide sequences can be easily produced from nucleotide precursors in a template-free non-enzymatic way under dehydrating conditions, i.e. using essentially dry materials. Here we report that 3’,5’ cyclic AMP may also serve as a substrate of the reaction, which proceeds under moderate conditions yet with a lower efficiency than the previously reported oligomerization of 3’,5’ cyclic GMP. Optimally the oligomerization requires (i) a temperature of 80°C, (ii) a neutral to alkaline environment and (iii) a time on the order of weeks. Differences in the yield and required reaction conditions of the oligomerizations utilizing 3’,5’ cGMP and cAMP are discussed in terms of the crystal structures of the compounds. Polymerization of 3’,5’ cyclic nucleotides, whose paramount relevance in a prebiotic chemistry context has been widely accepted for decades, supports the possibility that the origin of extant genetic materials might have followed a direct uninterrupted path since its very beginning, starting from non-elaborately pre-activated monomer compounds and simple reactions. PMID:27802310

  2. Changes in dynamics upon oligomerization regulate substrate binding and allostery in amino acid kinase family members.

    Directory of Open Access Journals (Sweden)

    Enrique Marcos

    2011-09-01

    Full Text Available Oligomerization is a functional requirement for many proteins. The interfacial interactions and the overall packing geometry of the individual monomers are viewed as important determinants of the thermodynamic stability and allosteric regulation of oligomers. The present study focuses on the role of the interfacial interactions and overall contact topology in the dynamic features acquired in the oligomeric state. To this aim, the collective dynamics of enzymes belonging to the amino acid kinase family both in dimeric and hexameric forms are examined by means of an elastic network model, and the softest collective motions (i.e., lowest frequency or global modes of motions favored by the overall architecture are analyzed. Notably, the lowest-frequency modes accessible to the individual subunits in the absence of multimerization are conserved to a large extent in the oligomer, suggesting that the oligomer takes advantage of the intrinsic dynamics of the individual monomers. At the same time, oligomerization stiffens the interfacial regions of the monomers and confers new cooperative modes that exploit the rigid-body translational and rotational degrees of freedom of the intact monomers. The present study sheds light on the mechanism of cooperative inhibition of hexameric N-acetyl-L-glutamate kinase by arginine and on the allosteric regulation of UMP kinases. It also highlights the significance of the particular quaternary design in selectively determining the oligomer dynamics congruent with required ligand-binding and allosteric activities.

  3. Differential structural remodelling of heparan sulfate by chemokines: the role of chemokine oligomerization

    Science.gov (United States)

    Migliorini, Elisa; Salanga, Catherina L.; Thakar, Dhruv

    2017-01-01

    Chemokines control the migration of cells in normal physiological processes and in the context of disease such as inflammation, autoimmunity and cancer. Two major interactions are involved: (i) binding of chemokines to chemokine receptors, which activates the cellular machinery required for movement; and (ii) binding of chemokines to glycosaminoglycans (GAGs), which facilitates the organization of chemokines into haptotactic gradients that direct cell movement. Chemokines can bind and activate their receptors as monomers; however, the ability to oligomerize is critical for the function of many chemokines in vivo. Chemokine oligomerization is thought to enhance their affinity for GAGs, and here we show that it significantly affects the ability of chemokines to accumulate on and be retained by heparan sulfate (HS). We also demonstrate that several chemokines differentially rigidify and cross-link HS, thereby affecting HS rigidity and mobility, and that HS cross-linking is significantly enhanced by chemokine oligomerization. These findings suggest that chemokine–GAG interactions may play more diverse biological roles than the traditional paradigms of physical immobilization and establishment of chemokine gradients; we hypothesize that they may promote receptor-independent events such as physical re-organization of the endothelial glycocalyx and extracellular matrix, as well as signalling through proteoglycans to facilitate leukocyte adhesion and transmigration. PMID:28123055

  4. Oligomerization of Clostridium perfringens epsilon toxin is dependent upon caveolins 1 and 2.

    Directory of Open Access Journals (Sweden)

    Christine M Fennessey

    Full Text Available Evidence from multiple studies suggests that Clostridium perfringens ε-toxin is a pore-forming toxin, assembling into oligomeric complexes in the plasma membrane of sensitive cells. In a previous study, we used gene-trap mutagenesis to identify mammalian factors contributing to toxin activity, including caveolin-2 (CAV2. In this study, we demonstrate the importance of caveolin-2 and its interaction partner, caveolin-1 (CAV1, in ε-toxin-induced cytotoxicity. Using CAV2-specific shRNA in a toxin-sensitive human kidney cell line, ACHN, we confirmed that cells deficient in CAV2 exhibit increased resistance to ε-toxin. Similarly, using CAV1-specific shRNA, we demonstrate that cells deficient in CAV1 also exhibit increased resistance to the toxin. Immunoprecipitation of CAV1 and CAV2 from ε-toxin-treated ACHN cells demonstrated interaction of both CAV1 and -2 with the toxin. Furthermore, blue-native PAGE indicated that the toxin and caveolins were components of a 670 kDa protein complex. Although ε-toxin binding was only slightly perturbed in caveolin-deficient cells, oligomerization of the toxin was dramatically reduced in both CAV1- and CAV2-deficient cells. These results indicate that CAV1 and -2 potentiate ε-toxin induced cytotoxicity by promoting toxin oligomerization - an event which is requisite for pore formation and, by extension, cell death.

  5. Phosphomimetic Mutations Enhance Oligomerization of Phospholemman and Modulate Its Interaction with the Na/K-ATPase*

    Science.gov (United States)

    Song, Qiujing; Pallikkuth, Sandeep; Bossuyt, Julie; Bers, Donald M.; Robia, Seth L.

    2011-01-01

    Na/K-ATPase (NKA) activity is dynamically regulated by an inhibitory interaction with a small transmembrane protein, phospholemman (PLM). Inhibition is relieved upon PLM phosphorylation. Phosphorylation may alter how PLM interacts with NKA and/or itself, but details of these interactions are unknown. To address this, we quantified FRET between PLM and its regulatory target NKA in live cells. Phosphorylation of PLM was mimicked by mutation S63E (PKC site), S68E (PKA/PKC site), or S63E/S68E. The dependence of FRET on protein expression in live cells yielded information about the structure and binding affinity of the PLM-NKA regulatory complex. PLM phosphomimetic mutations altered the quaternary structure of the regulatory complex and reduced the apparent affinity of the PLM-NKA interaction. The latter effect was likely due to increased oligomerization of PLM phosphomimetic mutants, as suggested by PLM-PLM FRET measurements. Distance constraints obtained by FRET suggest that phosphomimetic mutations slightly alter the oligomer quaternary conformation. Photon-counting histogram measurements revealed that the major PLM oligomeric species is a tetramer. We conclude that phosphorylation of PLM increases its oligomerization into tetramers, decreases its binding to NKA, and alters the structures of both the tetramer and NKA regulatory complex. PMID:21220422

  6. Disruption of oligomerization induces nucleocytoplasmic shuttling of leukemia-associated rho Guanine-nucleotide exchange factor.

    Science.gov (United States)

    Grabocka, Elda; Wedegaertner, Philip B

    2007-10-01

    The rgsRhoGEFs comprise a subfamily of three guanine nucleotide exchange factors, which function in linking heterotrimeric G-proteins to the monomeric RhoGTPase. Here, we reveal the novel finding that oligomerization of leukemia-associated RhoGEF (LARG) functions to prevent nucleocytoplasmic shuttling and to retain LARG in the cytoplasm. We establish that oligomerization is mediated by a predicted coiled-coil sequence (amino acids 1507-1520) in the extreme C terminus of LARG and that substitution of isoleucines 1507/1510 with alanines disrupts homo-oligomerization and leads to nucleocytoplasmic shuttling via the CRM1 nuclear transport pathway. In addition, we demonstrate that induced dimerization of an otherwise nuclear monomeric LARG mutant promotes cytoplasmic localization. Furthermore, we establish that nuclear import of monomeric LARG is mediated by the nuclear localization sequence (29)PTDKKQK(35) in the extreme N terminus. We propose that nucleocytoplasmic shuttling provides a mechanism for spatially regulating the activity of LARG toward its cytoplasmic targets and potentially new nuclear targets.

  7. Organization of the mitochondrial apoptotic BAK pore: oligomerization of the BAK homodimers.

    Science.gov (United States)

    Aluvila, Sreevidya; Mandal, Tirtha; Hustedt, Eric; Fajer, Peter; Choe, Jun Yong; Oh, Kyoung Joon

    2014-01-31

    The multidomain pro-apoptotic Bcl-2 family proteins BAK and BAX are believed to form large oligomeric pores in the mitochondrial outer membrane during apoptosis. Formation of these pores results in the release of apoptotic factors including cytochrome c from the intermembrane space into the cytoplasm, where they initiate the cascade of events that lead to cell death. Using the site-directed spin labeling method of electron paramagnetic resonance (EPR) spectroscopy, we have determined the conformational changes that occur in BAK when the protein targets to the membrane and forms pores. The data showed that helices α1 and α6 disengage from the rest of the domain, leaving helices α2-α5 as a folded unit. Helices α2-α5 were shown to form a dimeric structure, which is structurally homologous to the recently reported BAX "BH3-in-groove homodimer." Furthermore, the EPR data and a chemical cross-linking study demonstrated the existence of a hitherto unknown interface between BAK BH3-in-groove homodimers in the oligomeric BAK. This novel interface involves the C termini of α3 and α5 helices. The results provide further insights into the organization of the BAK oligomeric pores by the BAK homodimers during mitochondrial apoptosis, enabling the proposal of a BAK-induced lipidic pore with the topography of a "worm hole."

  8. Dealuminated ZSM-5 Zeolite Catalyst for Ethylene Oligomerization to Liquid Fuels

    Institute of Scientific and Technical Information of China (English)

    Nor Aishah Saidina Amin; Didi Dwi Anggoro

    2002-01-01

    Ethylene oligomerization using ZSM-5 zeolite was investigated to study the role of Bronstedacid sites in the formation of higher hydrocarbons. The oligomerization of olefins, dependent on the acidityof ZSM-5 zeolite, is an important step in the conversion of natural gas to liquid fuels. The framework Si/Alratio reflects the number of potential acid sites and the acid strength of the ZSM-5 catalyst. ZSM-5 withthe mole ratio SiO2/Al2O3 equal to 30 was dealuminated for different periods of time according to theacidic ion-exchange method to produce ZSM-5 with various Si/Al ratios. The FT-IR analysis revealedthat the integrated framework aluminum band, non-framework aluminum band, and silanol groups areasof the ZSM-5 zeolites decreased after being dealuminated. The performance of the dealuminated zeolitewas tested for ethylene oligomerization. The results demonstrated that the dealumination of ZSM-5 ledto higher ethylene conversion, but the gasoline selectivity was reduced compared to the performance of aZSM-5 zeolite. The characterization results revealed the amount of aluminum in the zeolitic framework,the crystallinity of the ZSM-5 zeolite, and the Si/Al ratio affected the formation of Bronsted acid sites.The number of the Bronsted acid sites on the catalyst active sites is important in the olefin conversion toliquid hydrocarbons.

  9. poolMC: Smart pooling of mRNA samples in microarray experiments

    Directory of Open Access Journals (Sweden)

    Schiefelbein John

    2010-06-01

    Full Text Available Abstract Background Typically, pooling of mRNA samples in microarray experiments implies mixing mRNA from several biological-replicate samples before hybridization onto a microarray chip. Here we describe an alternative smart pooling strategy in which different samples, not necessarily biological replicates, are pooled in an information theoretic efficient way. Further, each sample is tested on multiple chips, but always in pools made up of different samples. The end goal is to exploit the compressibility of microarray data to reduce the number of chips used and increase the robustness to noise in measurements. Results A theoretical framework to perform smart pooling of mRNA samples in microarray experiments was established and the software implementation of the pooling and decoding algorithms was developed in MATLAB. A proof-of-concept smart pooled experiment was performed using validated biological samples on commercially available gene chips. Differential-expression analysis of the smart pooled data was performed and compared against the unpooled control experiment. Conclusions The theoretical developments and experimental demonstration in this paper provide a useful starting point to investigate smart pooling of mRNA samples in microarray experiments. Although the smart pooled experiment did not compare favorably with the control, the experiment highlighted important conditions for the successful implementation of smart pooling - linearity of measurements, sparsity in data, and large experiment size.

  10. SAFETY AND MANAGEMENT OF SWIMING POOLS

    Directory of Open Access Journals (Sweden)

    Cemal GÜNDOĞDU

    2008-04-01

    Full Text Available In this study,it was investigated the situation related to the safety and management of swimming pools belongs to municipality This study was planned to determine the present situation related to the safety and management of open,half-olympic and closed swimming pools which are especially used for sports activities,to find out the deficiencies in practise and to overcome these.Our study included totally 80 open,closed,olympic, half-olympic public swimming pools(the pools that belongs to municipality,private administration and university,the colleges connected to the ministry of education, Military schools and police college.In our research,search,natural observation and meeting methods were used.In accordance with this aim the checklist questions,which were prepared for health and safety controls in swimming pools, were tested on the people by face-to-face meeting method. While the statistical evaluation of the available results were being done,frequency and percentage dispersion obtained from checklist was found.It was found that the 52 % of the training and lifeguard choise of the pools in Turkey,64 % of the emergency action equipment,71 % of the signs and signboards for the safety of the pool,75 % of the pool edge and its surroundings are not suitable for the standards and that these threaten the swimmers health importantly.Consequently,we are in the opinion that the pool staff should receive the periodical training about the first aid and using emergency situation equipment.And it is necessary that the Ministry Responsible for sports,Ministry of Interior,Ministry of Health,Ministry of Tourism be cooperate with water sports federations and the related civil society organizationsKey Words: .

  11. LCG POOL development status and production experience

    CERN Document Server

    Chytracek, Radovan; Cioffi, Carmine; Düllmann, Dirk; Frank, Markus; Girone, Maria; Govi, Giacomo; Karr, Kristo; Malon, David; Moscicki, Jakub T; Papadopoulos, Ioannis M; Schmücker, H; Tanenbaum, William; Vaniachine, A; Xie Zhen

    2004-01-01

    The POOL project, as a part of the LHC Computing Grid (LCG), is now entering its third year of active development POOL provides the baseline persistency framework for three LHC experiment and is based on a strict component model, insulating experiment software from a variety of storage technology choices. This paper gives a brief overview of the POOL architecture, its main design principles and the experience gained with integration into LHC experiment frameworks. In also presents recent developments in the area of relational database abstraction and object storage into RDBMS systems.

  12. Airways disorders and the swimming pool.

    Science.gov (United States)

    Bougault, Valérie; Boulet, Louis-Philippe

    2013-08-01

    Concerns have been expressed about the possible detrimental effects of chlorine derivatives in indoor swimming pool environments. Indeed, a controversy has arisen regarding the possibility that chlorine commonly used worldwide as a disinfectant favors the development of asthma and allergic diseases. The effects of swimming in indoor chlorinated pools on the airways in recreational and elite swimmers are presented. Recent studies on the influence of swimming on airway inflammation and remodeling in competitive swimmers, and the phenotypic characteristics of asthma in this population are reviewed. Preventative measures that could potentially reduce the untoward effects of pool environment on airways of swimmers are discussed. Copyright © 2013 Elsevier Inc. All rights reserved.

  13. Spectrum pooling in MnWave Networks

    DEFF Research Database (Denmark)

    Boccardi, Federico; Shokri-Ghadikolaei, Hossein; Fodor, Gabor

    2016-01-01

    the benefit of coordination among networks of different operators, study the impact of beamforming at both base stations and user terminals, and analyze the pooling performance at different frequency carriers. We also discuss the enabling spectrum mechanisms, architectures, and protocols required to make...... spectrum pooling work in real networks. Our initial results show that, from a technical perspective, spectrum pooling at mmWave has the potential to use the resources more efficiently than traditional exclusive spectrum allocation to a single operator. However, further studies are needed in order to reach...... a thorough understanding of this matter, and we hope that this article will help stimulate further research in this area....

  14. Weld pool visual sensing without external illumination

    DEFF Research Database (Denmark)

    Liu, Jinchao; Fan, Zhun; Olsen, Soren Ingvor

    2011-01-01

    Visual sensing in arc welding has become more and more important, but still remains challenging because of the harsh environment with extremely strong illumination from the arc. This paper presents a low-cost camera-based sensor system, without using external Illumination, but nevertheless able...... to sense and model the weld pool. Central is a carefully selected optical filtering as well as an active contour-based tracking of the weld pool boundary. The system is able to extract the 2D shape of the weld pool in real time. The reported experiments show the feasibility of this approach....

  15. 10 CFR 36.63 - Pool water purity.

    Science.gov (United States)

    2010-01-01

    ... § 36.63 Pool water purity. (a) Pool water purification system must be run sufficiently to maintain the conductivity of the pool water below 20 microsiemens per centimeter under normal circumstances. If pool water... 10 Energy 1 2010-01-01 2010-01-01 false Pool water purity. 36.63 Section 36.63 Energy...

  16. Microbial quality of a marine tidal pool

    CSIR Research Space (South Africa)

    Genthe, Bettina

    1995-01-01

    Full Text Available In this study the source of microbial pollution to a tidal pool was investigated. Both adjacent seawater which could contribute to possible faecal pollution and potential direct bather pollution were studied. The microbial quality of the marine...

  17. Ingestion of swimming pool water by recreational

    Data.gov (United States)

    U.S. Environmental Protection Agency — Swimming pool water ingestion data. This dataset is associated with the following publication: Dufour, A., L. Wymer, M. Magnuson, T. Behymer, and R. Cantu. Ingestion...

  18. Tehas teisel pool Kuud / Gert Kiiler

    Index Scriptorium Estoniae

    Kiiler, Gert

    2009-01-01

    Tartu Ülikooli Viljandi kultuuriakadeemia teatrikunsti tudengite seitsmendast lennust kõneleva saate "Tehas teisel pool Kuud" tegemisest. Saade Eesti Televisioonis 3. juunil. Produtsent Kadi Katarina Priske, režissöör Elo Selirand

  19. AE/VCE Confirmed Vernal Pools

    Data.gov (United States)

    Vermont Center for Geographic Information — This dataset is derived from a project by the Vermont Center for Ecostudies(VCE) and Arrowwood Environmental(AE) to map vernal pools throughout the state of Vermont....

  20. Robotic cleaning of a spent fuel pool

    Energy Technology Data Exchange (ETDEWEB)

    Roman, H.T.; Marian, F.A. (PSE and G Research Corp., Newark, NJ (US)); Silverman, E.B.; Barkley, V.P. (ARD Corp., Columbia, MD (US))

    1987-05-01

    Spent fuel pools at nuclear power plants are not cleaned routinely, other than by purifying the water that they contain. Yet, debris can collect on the bottom of a pool and should be removed prior to fuel transfer. At Public Service Electric and Gas Company's Hope Creek Nuclear Power Plant, a submersible mobile robot - ARD Corporation's SCAVENGER - was used to clean the bottom of the spent fuel pool prior to initial fuel loading. The robotic device was operated remotely (as opposed to autonomously) with a simple forward/reverse control, and it cleaned 70-80% of the pool bottom. This paper reports that a simple cost-benefit analysis shows that the robotic device would be less expensive, on a per mission basis, than other cleaning alternatives, especially if it were used for other similar cleaning operations throughout the plant.

  1. Analysis of Bitcoin Pooled Mining Reward Systems

    CERN Document Server

    Rosenfeld, Meni

    2011-01-01

    In this paper we describe the various scoring systems used to calculate rewards of participants in Bitcoin pooled mining, explain the problems each were designed to solve and analyze their respective advantages and disadvantages.

  2. How to map your industry's profit pool.

    Science.gov (United States)

    Gadiesh, O; Gilbert, J L

    1998-01-01

    Many managers chart strategy without a full understanding of the sources and distribution of profits in their industry. Sometimes they focus their sights on revenues instead of profits, mistakenly assuming that revenue growth will eventually translate into profit growth. In other cases, they simply lack the data or the analytical tools required to isolate and measure variations in profitability. In this Manager's Tool Kit, the authors present a way to think clearly about where the money's being made in any industry. They describe a framework for analyzing how profits are distributed among the activities that form an industry's value chain. Such an analysis can provide a company's managers with a rich understanding of their industry's profit structure--what the authors call its profit pool--enabling them to identify which activities are generating disproportionately large or small shares of profits. Even more important, a profit-pool map opens a window onto the underlying structure of the industry, helping managers see the various forces that are determining the distribution of profits. As such, a profit-pool map provides a solid basis for strategic thinking. Mapping a profit pool involves four steps: defining the boundaries of the pool, estimating the pool's overall size, estimating the size of each value-chain activity in the pool, and checking and reconciling the calculations. The authors briefly describe each step and then apply the process by providing a detailed example of a hypothetical retail bank. They conclude by looking at ways of organizing the data in chart form as a first step toward plotting a profit-pool strategy.

  3. Pooling strategies for St Petersburg gamblers

    OpenAIRE

    Csörgö, Sandor; Simons, Gordon

    2006-01-01

    Peter offers to play exactly one St Petersburg game with each of [math] players, Paul [math] , [math] , Paul [math] , whose conceivable pooling strategies are described by all possible probability distributions [math] . Comparing infinite expectations, we characterize among all [math] those admissible strategies for which the pooled winnings, each distributed as [math] , yield a finite added value for each and every one of Paul [math] , [math] , Paul [math] in comparison with their individual...

  4. Performance Study of Swimming Pool Heaters

    Energy Technology Data Exchange (ETDEWEB)

    McDonald, R.J.

    2009-01-01

    The objective of this report is to perform a controlled laboratory study on the efficiency and emissions of swimming pool heaters based on a limited field investigation into the range of expected variations in operational parameters. Swimming pool heater sales trends have indicated a significant decline in the number of conventional natural gas-fired swimming pool heaters (NGPH). On Long Island the decline has been quite sharp, on the order of 50%, in new installations since 2001. The major portion of the decline has been offset by a significant increase in the sales of electric powered heat pump pool heaters (HPPH) that have been gaining market favor. National Grid contracted with Brookhaven National Laboratory (BNL) to measure performance factors in order to compare the relative energy, environmental and economic consequences of using one technology versus the other. A field study was deemed inappropriate because of the wide range of differences in actual load variations (pool size), geographic orientations, ground plantings and shading variations, number of hours of use, seasonal use variations, occupancy patterns, hour of the day use patterns, temperature selection, etc. A decision was made to perform a controlled laboratory study based on a limited field investigation into the range of expected operational variations in parameters. Critical to this are the frequency of use, temperature selection, and sizing of the heater to the associated pool heating loads. This would be accomplished by installing a limited amount of relatively simple compact field data acquisition units on selected pool installations. This data included gas usage when available and alternately heater power or gas consumption rates were inferred from the manufacturer's specifications when direct metering was not available in the field. Figure 1 illustrates a typical pool heater installation layout.

  5. Profit pools: a fresh look at strategy.

    Science.gov (United States)

    Gadiesh, O; Gilbert, J L

    1998-01-01

    In charting strategy, many managers focus on revenue growth, assuming that profits will follow. But that approach is dangerous: today's deep revenue pool may become tomorrow's dry hole. To create strategies that result in profitable growth, managers need to look beyond revenues to see the shape of their industry's profit pool. The authors define an industry's profit pool as the total profits earned at all points along the industry's value chain. Although the concept is simple, the structure of a profit pool is usually quite complex. The pool will be deeper in some segments of the value chain than in others, and depths will vary within an individual segment as well. Segment profitability may, for example, vary widely by customer group, product category, geographic market, and distribution channel. Moreover, the pattern of profit concentration in an industry will often be very different from the pattern of revenue concentration. The authors describe how successful companies have gained competitive advantage by developing sophisticated profit-pool strategies. They explain how U-Haul identified new sources of profit in the consumer-truck-rental industry; how Merck reached beyond its traditional value-chain role to protect its profits in the pharmaceuticals industry; how Dell rebounded from a misguided channel decision by refocusing on its traditional source of profit; and how Anheuser-Busch made a series of astute product, pricing, and operating decisions to dominate the beer industry's profit pool. The companies with the best understanding of their industry's profit pool, the authors argue, will be in the best position to thrive over the long term.

  6. Pooling strategies for St Petersburg gamblers

    OpenAIRE

    Csörgö, Sandor; Simons, Gordon

    2006-01-01

    Peter offers to play exactly one St Petersburg game with each of [math] players, Paul [math] , [math] , Paul [math] , whose conceivable pooling strategies are described by all possible probability distributions [math] . Comparing infinite expectations, we characterize among all [math] those admissible strategies for which the pooled winnings, each distributed as [math] , yield a finite added value for each and every one of Paul [math] , [math] , Paul [math] in comparison with their individual...

  7. Welding pool measurement using thermal array sensor

    Science.gov (United States)

    Cho, Chia-Hung; Hsieh, Yi-Chen; Chen, Hsin-Yi

    2015-08-01

    Selective laser melting (SLM) is an additive manufacturing (AM) technology that uses a high-power laser beam to melt metal powder in chamber of inert gas. The process starts by slicing the 3D CAD data as a digital information source into layers to create a 2D image of each layer. Melting pool was formed by using laser irradiation on metal powders which then solidified to consolidated structure. In a selective laser melting process, the variation of melt pool affects the yield of a printed three-dimensional product. For three dimensional parts, the border conditions of the conductive heat transport have a very large influence on the melt pool dimensions. Therefore, melting pool is an important behavior that affects the final quality of the 3D object. To meet the temperature and geometry of the melting pool for monitoring in additive manufacturing technology. In this paper, we proposed the temperature sensing system which is composed of infrared photodiode, high speed camera, band-pass filter, dichroic beam splitter and focus lens. Since the infrared photodiode and high speed camera look at the process through the 2D galvanometer scanner and f-theta lens, the temperature sensing system can be used to observe the melting pool at any time, regardless of the movement of the laser spot. In order to obtain a wide temperature detecting range, 500 °C to 2500 °C, the radiation from the melting pool to be measured is filtered into a plurality of radiation portions, and since the intensity ratio distribution of the radiation portions is calculated by using black-body radiation. The experimental result shows that the system is suitable for melting pool to measure temperature.

  8. Recent advances in probabilistic species pool delineations

    Directory of Open Access Journals (Sweden)

    Dirk Nikolaus Karger

    2016-07-01

    Full Text Available A species pool is the set of species that could potentially colonize and establish within a community. It has been a commonly used concept in biogeography since the early days of MacArthur and Wilson’s work on Island Biogeography. Despite their simple and appealing definition, an operational application of species pools is bundled with a multitude of problems, which have often resulted in arbitrary decisions and workarounds when defining species pools. Two recently published papers address the operational problems of species pool delineations, and show ways of delineating them in a probabilistic fashion. In both papers, species pools were delineated using a process-based, mechanistical approach, which opens the door for a multitude of new applications in biogeography. Such applications include detecting the hidden signature of biotic interactions, disentangling the geographical structure of community assembly processes, and incorporating a temporal extent into species pools. Although similar in their conclusions, both ‘probabilistic approaches’ differ in their implementation and definitions. Here I give a brief overview of the differences and similarities of both approaches, and identify the challenges and advantages in their application.

  9. A Review of Selected International Aircraft Spares Pooling Programs: Lessons Learned for F-35 Spares Pooling

    Science.gov (United States)

    2016-01-01

    international military aircraft support program using true spares pooling for which we have any detailed and reliable data, and it is the closest current...for all F77 spares . The remaining non-C-17 unique spares are managed by the Defense Logistics Agency and the Air Force Air Logistics Centers .50...C O R P O R A T I O N Research Report A Review of Selected International Aircraft Spares Pooling Programs Lessons Learned for F-35 Spares Pooling

  10. Stade NPP. Dismantling of the reactor pool

    Energy Technology Data Exchange (ETDEWEB)

    Scharf, Daniel; Dziwis, Joachim [E.ON Anlagenservice GmbH Nukleartechnik, Gelsenkirchen (Germany); Kemp, Lutz-Hagen [KKW Stade GmbH und Co. oHG, Stade (Germany)

    2012-11-01

    Within the scope of the 4{sup th} partial decommissioning permission of Stade NPP the activated and contaminated structures of the reactor pool had to be dismantled in order to gain a completely non-radioactive reactor pool area for the subsequent clearance measurement of the reactor building. In order to achieve the aim it was intended to remove the activated pool liner sheets, its activated framework and several contaminated ventilation channels made of stainless steel, the concrete walls of the reactor pool entirely or in parts depending on their activation level, as well as the remaining activated carbon steel structures of the reactor pool bottom. Embedded in the concrete walls there were several highly contaminated excore tubes and the contaminated pool top edge, which were intended to be removed to its full extent. The contract of the Stade NPP initiated reactor pool dismantling project had been awarded to E.ON Anlagenservice GmbH (EAS) and its subsupplier sat. Kerntechnik GmbH for the concrete dismantling works and was performed as follows. In order to minimize the radiation level in the main working area in accordance with the ALARA principle, the liner sheets and middle parts of its framework were removed by means of angle grinders first, as they were the most dose rate relevant parts. As a result the primary average radiation level in the reactor pool (measured in a distance of 500 mm from the walls) was lowered from 40 {mu}Sv/h to less than 2 {mu}Sv/h. After the minimization of the radiation level in the working area the main dismantling step started with the cutting of the reactor pool walls in blocks by means of diamond rope cutters. Once a concrete block was cut out, it was transported into the fuel pool by means of a crane and crane fork, examined radiologically, marked area by area and segmented to debris by means of an electrical excavator with a hydraulic chisel. Afterwards the debris and carbon steel parts were fractioned and packed for further

  11. Vernal Pool Distribution - Central Valley, 2005 [ds650

    Data.gov (United States)

    California Department of Resources — "Great Valley Vernal Pool Distribution", originally mapped by Bob Holland, 2005. This dataset contains vernal pool areas mapped over Califorina's Central Valley,...

  12. Alkali metal pool boiler life tests for a 25 kWe advanced Stirling conversion system

    Science.gov (United States)

    Anderson, W. G.; Rosenfeld, J. H.; Noble, J.

    The overall operating temperature and efficiency of solar-powered Stirling engines can be improved by adding an alkali metal pool boiler heat transport system to supply heat more uniformly to the heater head tubes. One issue with liquid metal pool boilers is unstable boiling. Stable boiling is obtained with an enhanced boiling surface containing nucleation sites that promote continuous boiling. Over longer time periods, it is possible that the boiling behavior of the system will change. An 800-h life test was conducted to verify that pool boiling with the chosen fluid/surface combination remains stable as the system ages. The apparatus uses NaK boiling on a - 100 + 140 stainless steel sintered porous layer, with the addition of a small amount of xenon. Pool boiling remained stable to the end of life test. The pool boiler life test included a total of 82 cold starts, to simulate startup each morning, and 60 warm restarts, to simulate cloud cover transients. The behavior of the cold and warm starts showed no significant changes during the life test. In the experiments, the fluid/surface combination provided stable, high-performance boiling at the operating temperature of 700 C. Based on these experiments, a pool boiler was designed for a full-scale 25-kWe Stirling system.

  13. Evaluation of the pool chemistry model in MELCOR by considering the organic iodine formation within the pool using the phebus FPT-1 test data

    Energy Technology Data Exchange (ETDEWEB)

    Park, Jong Hwa; Kim, Dong Ha [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of)

    2004-07-01

    In the severe accident, the understanding on the chemical form of iodine and its behavior is important for estimating the source terms because it has not only a active chemical features but also hazard to the public. From the current understanding on the early phase of severe accident, it is known that the most of the injected iodine into the containment is the aerosol form of metal iodide such as cesium iodide and silver iodide. The remaining fraction is the gaseous form of iodine such as molecular iodine, elemental iodine or hydrogen iodide. These aerosol and gaseous iodine can be removed by the engineered safety features such as the spray system. Also the natural depletion processes like condensing, settling or deposit on the surface remove the most of the suspended aerosol and gaseous iodine and finally, cause them to accumulate within the pool on the bottom floor. As time goes on, in the late phase of severe accident, the iodine pool chemistry phenomena starts under the strong radiation condition. The dissolved iodine within the pool can be transformed to the volatile iodine such as molecular iodine and organic iodine due to its strong chemical activity and various kind of immersed impurities such as alcohol or grease. This transformed volatile iodine can be partitioned to the atmosphere from the inside of pool. Consequently, an equilibrium concentration is reached and this suspended gaseous form of iodine becomes the main threat to the public when the containment fails or leaks in the late phase of severe accident. Therefore, decreasing the equilibrium concentration of the suspended gaseous iodine and the trapping of iodine within the pool are the key parameters of the iodine management strategy in the late phase of severe accident. The Phebus FPT-1 showed that the silver could play an important role in trapping the iodine in the sump by forming the insoluble silver iodide from I{sub 2} or I. The formation of volatile iodine depends on the PH, pool

  14. Swimming pool use and birth defect risk.

    Science.gov (United States)

    Agopian, A J; Lupo, Philip J; Canfield, Mark A; Mitchell, Laura E

    2013-09-01

    Swimming during pregnancy is recommended. However, the use of swimming pools is also associated with infection by water-borne pathogens and exposure to water disinfection byproducts, which are 2 mechanisms that are suspected to increase risk for birth defects. Thus, we evaluated the relationship between maternal swimming pool use during early pregnancy and risk for select birth defects in offspring. Data were evaluated for nonsyndromic cases with 1 of 16 types of birth defects (n = 191-1829) and controls (n = 6826) from the National Birth Defects Prevention Study delivered during 2000-2006. Logistic regression analyses were conducted separately for each birth defect type. Separate analyses were conducted to assess any pool use (yes vs no) and frequent use (5 or more occasions in 1 month) during the month before pregnancy through the third month of pregnancy. There was no significant positive association between any or frequent pool use and any of the types of birth defects, even after adjustment for several potential confounders (maternal race/ethnicity, age at delivery, education, body mass index, folic acid use, nulliparity, smoking, annual household income, surveillance center, and season of conception). Frequent pool use was significantly negatively associated with spina bifida (adjusted odds ratio, 0.68; 95% confidence interval, 0.47-0.99). Among offspring of women 20 years old or older, pool use was associated with gastroschisis (adjusted odds ratio, 1.3; 95% confidence interval, 1.0-1.8), although not significantly so. We observed little evidence suggesting teratogenic effects of swimming pool use. Because swimming is a common and suggested form of exercise during pregnancy, these results are reassuring. Copyright © 2013 Mosby, Inc. All rights reserved.

  15. There's plenty of gloom at the bottom: the many challenges of accurate quantitation in size-based oligomeric separations.

    Science.gov (United States)

    Striegel, André M

    2013-11-01

    There is a variety of small-molecule species (e.g., tackifiers, plasticizers, oligosaccharides) the size-based characterization of which is of considerable scientific and industrial importance. Likewise, quantitation of the amount of oligomers in a polymer sample is crucial for the import and export of substances into the USA and European Union (EU). While the characterization of ultra-high molar mass macromolecules by size-based separation techniques is generally considered a challenge, it is this author's contention that a greater challenge is encountered when trying to perform, for quantitation purposes, separations in and of the oligomeric region. The latter thesis is expounded herein, by detailing the various obstacles encountered en route to accurate, quantitative oligomeric separations by entropically dominated techniques such as size-exclusion chromatography, hydrodynamic chromatography, and asymmetric flow field-flow fractionation, as well as by methods which are, principally, enthalpically driven such as liquid adsorption and temperature gradient interaction chromatography. These obstacles include, among others, the diminished sensitivity of static light scattering (SLS) detection at low molar masses, the non-constancy of the response of SLS and of commonly employed concentration-sensitive detectors across the oligomeric region, and the loss of oligomers through the accumulation wall membrane in asymmetric flow field-flow fractionation. The battle is not lost, however, because, with some care and given a sufficient supply of sample, the quantitation of both individual oligomeric species and of the total oligomeric region is often possible.

  16. Hsp70 oligomerization is mediated by an interaction between the interdomain linker and the substrate-binding domain.

    Directory of Open Access Journals (Sweden)

    Francesco A Aprile

    Full Text Available Oligomerization in the heat shock protein (Hsp 70 family has been extensively documented both in vitro and in vivo, although the mechanism, the identity of the specific protein regions involved and the physiological relevance of this process are still unclear. We have studied the oligomeric properties of a series of human Hsp70 variants by means of nanoelectrospray ionization mass spectrometry, optical spectroscopy and quantitative size exclusion chromatography. Our results show that Hsp70 oligomerization takes place through a specific interaction between the interdomain linker of one molecule and the substrate-binding domain of a different molecule, generating dimers and higher-order oligomers. We have found that substrate binding shifts the oligomerization equilibrium towards the accumulation of functional monomeric protein, probably by sequestering the helical lid sub-domain needed to stabilize the chaperone: substrate complex. Taken together, these findings suggest a possible role of chaperone oligomerization as a mechanism for regulating the availability of the active monomeric form of the chaperone and for the control of substrate binding and release.

  17. Hsp70 Oligomerization Is Mediated by an Interaction between the Interdomain Linker and the Substrate-Binding Domain

    Science.gov (United States)

    Aprile, Francesco A.; Dhulesia, Anne; Stengel, Florian; Roodveldt, Cintia; Benesch, Justin L. P.; Tortora, Paolo; Robinson, Carol V.; Salvatella, Xavier; Dobson, Christopher M.; Cremades, Nunilo

    2013-01-01

    Oligomerization in the heat shock protein (Hsp) 70 family has been extensively documented both in vitro and in vivo, although the mechanism, the identity of the specific protein regions involved and the physiological relevance of this process are still unclear. We have studied the oligomeric properties of a series of human Hsp70 variants by means of nanoelectrospray ionization mass spectrometry, optical spectroscopy and quantitative size exclusion chromatography. Our results show that Hsp70 oligomerization takes place through a specific interaction between the interdomain linker of one molecule and the substrate-binding domain of a different molecule, generating dimers and higher-order oligomers. We have found that substrate binding shifts the oligomerization equilibrium towards the accumulation of functional monomeric protein, probably by sequestering the helical lid sub-domain needed to stabilize the chaperone: substrate complex. Taken together, these findings suggest a possible role of chaperone oligomerization as a mechanism for regulating the availability of the active monomeric form of the chaperone and for the control of substrate binding and release. PMID:23840795

  18. Comparative analyses of quaternary arrangements in homo-oligomeric proteins in superfamilies: Functional implications.

    Science.gov (United States)

    Sudha, Govindarajan; Srinivasan, Narayanaswamy

    2016-09-01

    A comprehensive analysis of the quaternary features of distantly related homo-oligomeric proteins is the focus of the current study. This study has been performed at the levels of quaternary state, symmetry, and quaternary structure. Quaternary state and quaternary structure refers to the number of subunits and spatial arrangements of subunits, respectively. Using a large dataset of available 3D structures of biologically relevant assemblies, we show that only 53% of the distantly related homo-oligomeric proteins have the same quaternary state. Considering these homologous homo-oligomers with the same quaternary state, conservation of quaternary structures is observed only in 38% of the pairs. In 36% of the pairs of distantly related homo-oligomers with different quaternary states the larger assembly in a pair shows high structural similarity with the entire quaternary structure of the related protein with lower quaternary state and it is referred as "Russian doll effect." The differences in quaternary state and structure have been suggested to contribute to the functional diversity. Detailed investigations show that even though the gross functions of many distantly related homo-oligomers are the same, finer level differences in molecular functions are manifested by differences in quaternary states and structures. Comparison of structures of biological assemblies in distantly and closely related homo-oligomeric proteins throughout the study differentiates the effects of sequence divergence on the quaternary structures and function. Knowledge inferred from this study can provide insights for improved protein structure classification and function prediction of homo-oligomers. Proteins 2016; 84:1190-1202. © 2016 Wiley Periodicals, Inc.

  19. Isolation and quantification of oligomeric and polymeric procyanidins in leaves and flowers of Hawthorn (Crataegus spp.).

    Science.gov (United States)

    Hellenbrand, N; Sendker, J; Lechtenberg, M; Petereit, F; Hensel, A

    2015-07-01

    Proanthocyanidins (PAs) constitute a class of polyphenols with flavan-3-ols as monomeric building blocks. These polyphenols are mostly quantified by colorimetric methods or by chromatographic determination of monomeric flavan-3-ols or low molecular oligomers as lead compounds. No reliable analytical methods are available for unambiguous identification of the homologues series of oligo- and polymeric PAs. For Hawthorn leaf and flower (Crataegi folium cum flore) from Crataegus spp. (Rosaceae) a protocol for preparative isolation of oligomeric and polymeric PAs from an acetone-water extract was developed, yielding procyanidin reference clusters with defined degree of polymerization (DP) from 2 to 10 besides a procyanidin-polymer. Identity and purity of these clusters were proven by HPLC, MS and in part NMR studies. For identification and quantification from Hawthorn an ICH-Q2 validated UHPLC method with fluorimetric detection and less than 10min runtime was developed. The method enabled quantification of procyanidin clusters with DP from 2 to 10 besides the polymer fraction. Batch analysis revealed procyanidin contents of about 20 to 45mg/g from a homologues series of oligomeric PAs and about 50% of polymer fraction. Monitoring of procyanidin distribution during seasonal growth of fresh plants of Crataegus monogyna showed more or less constant contents between 20 and 55mg/g dry weight of oligomeric procyanidins during the growing season in the different plant organs with strong accumulation in the flowers and fruits (55mg/g dry weight). From these data it can be speculated that procyanidins serve as part of the plants defense system in the reproductive organs of the plant. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. LDRD final report on new homogeneous and supported oligomerization catalysts (LDRD 42461).

    Energy Technology Data Exchange (ETDEWEB)

    Hascall, Anthony G.; Kemp, Richard Alan

    2004-11-01

    The overall purpose of this LDRD is multifold. First, we are interested in preparing new homogeneous catalysts that can be used in the oligomerization of ethylene and in understanding commercially important systems better. Second, we are interested in attempting to support these new homogeneous catalysts in the pores of nano- or mesoporous materials in order to force new and unusual distributions of a-olefins to be formed during the oligomerization. Thus the overall purpose is to try to prepare new catalytic species and to possibly control the active site architecture in order to yield certain desired products during a catalytic reaction, much like nature does with enzymes. In order to rationally synthesize catalysts it is imperative to comprehend the function of the various components of the catalyst. In heterogeneous systems, it is of utmost importance to know how a support interacts with the active site of the catalyst. In fact, in the catalysis world this lack of fundamental understanding of the relationship between active site and support is the single largest reason catalysis is considered an 'empirical' or 'black box' science rather than a well-understood one. In this work we will be preparing novel ethylene oligomerization catalysts, which are normally P-O chelated homogeneous complexes, with new ligands that replace P with a stable carbene. We will also examine a commercially catalyst system and investigate the active site in it via X-ray crystallography. We will also attempt to support these materials inside the pores of nano- and mesoporous materials. Essentially, we will be tailoring the size and scale of the catalyst active site and its surrounding environment to match the size of the molecular product(s) we wish to make. The overall purpose of the study will be to prepare new homogeneous catalysts, and if successful in supporting them to examine the effects that steric constraints and pore structures can have on growing oligomer

  1. The effects of oligomerization on Saccharomyces cerevisiae Mcm4/6/7 function

    Directory of Open Access Journals (Sweden)

    Davey Megan J

    2010-09-01

    Full Text Available Abstract Background Minichromosome maintenance proteins (Mcm 2, 3, 4, 5, 6 and 7 are related by sequence and form a variety of complexes that unwind DNA, including Mcm4/6/7. A Mcm4/6/7 trimer forms one half of the Mcm2-7 hexameric ring and can be thought of as the catalytic core of Mcm2-7, the replicative helicase in eukaryotic cells. Oligomeric analysis of Mcm4/6/7 suggests that it forms a hexamer containing two Mcm4/6/7 trimers, however, under certain conditions trimeric Mcm4/6/7 has also been observed. The functional significance of the different Mcm4/6/7 oligomeric states has not been assessed. The results of such an assessment would have implications for studies of both Mcm4/6/7 and Mcm2-7. Results Here, we show that Saccharomyces cerevisiae Mcm4/6/7 reconstituted from individual subunits exists in an equilibrium of oligomeric forms in which smaller oligomers predominate in the absence of ATP. In addition, we found that ATP, which is required for Mcm4/6/7 activity, shifts the equilibrium towards larger oligomers, likely hexamers of Mcm4/6/7. ATPγS and to a lesser extent ADP also shift the equilibrium towards hexamers. Study of Mcm4/6/7 complexes containing mutations that interfere with the formation of inter-subunit ATP sites (arginine finger mutants indicates that full activity of Mcm4/6/7 requires all of its ATP sites, which are formed in a hexamer and not a trimer. In keeping with this observation, Mcm4/6/7 binds DNA as a hexamer. Conclusions The minimal functional unit of Mcm4/6/7 is a hexamer. One of the roles of ATP binding by Mcm4/6/7 may be to stabilize formation of hexamers.

  2. Structural properties of cyanase. Denaturation, renaturation, and role of sulfhydryls and oligomeric structure in catalytic activity.

    Science.gov (United States)

    Little, R M; Anderson, P M

    1987-07-25

    Cyanase is an inducible enzyme in Escherichia coli that catalyzes bicarbonate-dependent decomposition of cyanate to give ammonia and bicarbonate. The enzyme is composed of 8-10 identical subunits (Mr = 17,008). The objective of this study was to clarify some of the structural properties of cyanase for the purpose of understanding the relationship between oligomeric structure and catalytic activity. Circular dichroism studies showed that cyanase has a significant amount of alpha-helix and beta-sheet structure. The one sulfhydryl group per subunit does not react with 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) unless cyanase is denatured. Denaturation is apparently complete in 10 M urea or 6 M guanidine hydrochloride, but is significantly reduced in 10 M urea by the presence of azide (analog of cyanate) and is incomplete in 8 M urea. Denatured cyanase could be renatured and reactivated (greater than 85%) by removal of denaturants. Reactivation was greatly facilitated by the presence of certain anions, particularly bicarbonate, and by high ionic strength and protein concentration. The catalytic activity of renatured cyanase was associated only with oligomer. Cyanase that had been denatured in the presence of DTNB to give a cyanase-DTNB derivative could also be renatured at 26 degrees C to give active cyanase-DTNB oligomer. The active oligomeric form of the cyanase-DTNB derivative could be converted reversibly to inactive dimer by lowering the temperature to 4 degrees C or by reduction of the ionic strength and removal of monoanions. These results provide evidence that free sulfhydryl groups are not required for catalytic activity and that catalytic activity may be dependent upon oligomeric structure.

  3. Mapping the conformational dynamics and pathways of spontaneous steric zipper Peptide oligomerization.

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    Dirk Matthes

    Full Text Available The process of protein misfolding and self-assembly into various, polymorphic aggregates is associated with a number of important neurodegenerative diseases. Only recently, crystal structures of several short peptides have provided detailed structural insights into -sheet rich aggregates, known as amyloid fibrils. Knowledge about early events of the formation and interconversion of small oligomeric states, an inevitable step in the cascade of peptide self-assembly, however, remains still limited. We employ molecular dynamics simulations in explicit solvent to study the spontaneous aggregation process of steric zipper peptide segments from the tau protein and insulin in atomistic detail. Starting from separated chains with random conformations, we find a rapid formation of structurally heterogeneous, -sheet rich oligomers, emerging from multiple bimolecular association steps and diverse assembly pathways. Furthermore, our study provides evidence that aggregate intermediates as small as dimers can be kinetically trapped and thus affect the structural evolution of larger oligomers. Alternative aggregate structures are found for both peptide sequences in the different independent simulations, some of which feature characteristics of the known steric zipper conformation (e.g., -sheet bilayers with a dry interface. The final aggregates interconvert with topologically distinct oligomeric states exclusively via internal rearrangements. The peptide oligomerization was analyzed through the perspective of a minimal oligomer, i.e., the dimer. Thereby all observed multimeric aggregates can be consistently mapped onto a space of reduced dimensionality. This novel method of conformational mapping reveals heterogeneous association and reorganization dynamics that are governed by the characteristics of peptide sequence and oligomer size.

  4. Oligomerization of Mannan-binding Lectin Dictates Binding Properties and Complement Activation.

    Science.gov (United States)

    Kjaer, T R; Jensen, L; Hansen, A; Dani, R; Jensenius, J C; Dobó, J; Gál, P; Thiel, S

    2016-07-01

    The complement system is a part of the innate immune system and is involved in recognition and clearance of pathogens and altered-self structures. The lectin pathway of the complement system is initiated when soluble pattern recognition molecules (PRMs) with collagen-like regions bind to foreign or altered self-surfaces. Associated with the collagen-like stems of these PRMs are three mannan-binding lectin (MBL)-associated serine proteases (MASPs) and two MBL-associated proteins (MAps). The most studied of the PRMs, MBL, is present in serum mainly as trimeric and tetrameric oligomers of the structural subunit. We hypothesized that oligomerization of MBL may influence both the potential to bind to micro organisms and the interaction with the MASPs and MAps, thus influencing the ability to initiate complement activation. When testing binding at 37 °C, we found higher binding of tetrameric MBL to Staphylococcus aureus (S. aureus) than trimeric and dimeric MBL. In serum, we found that tetrameric MBL was the main oligomeric form present in complexes with the MASPs and MAp44. Such preference was confirmed using purified forms of recombinant MBL (rMBL) oligomers, where tetrameric rMBL interacted stronger with all of the MASPs and MAp44, compared to trimeric MBL. As a direct consequence of the weaker interaction with the MASPs, we found that trimeric rMBL was inferior to tetrameric rMBL in activating the complement system. Our data suggest that the oligomeric state of MBL is crucial both for the binding properties and the effector function of MBL.

  5. APOBEC3G oligomerization is associated with the inhibition of both Alu and LINE-1 retrotransposition.

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    Takayoshi Koyama

    Full Text Available Alu and LINE-1 (L1, which constitute ~11% and ~17% of the human genome, respectively, are transposable non-LTR retroelements. They transpose not only in germ cells but also in somatic cells, occasionally causing cancer. We have previously demonstrated that antiretroviral restriction factors, human APOBEC3 (hA3 proteins (A-H, differentially inhibit L1 retrotransposition. In this present study, we found that hA3 members also restrict Alu retrotransposition at differential levels that correlate with those observed previously for L1 inhibition. Through deletion analyses based on the best-characterized hA3 member human APOBEC3G (hA3G, its N-terminal 30 amino acids were required for its inhibitory activity against Alu retrotransposition. The inhibitory effect of hA3G on Alu retrotransposition was associated with its oligomerization that was affected by the deletion of its N-terminal 30 amino acids. Through structural modeling, the amino acids 24 to 28 of hA3G were predicted to be located at the interface of the dimer. The mutation of these residues resulted in abrogated hA3G oligomerization, and consistently abolished the inhibitory activity of hA3G against Alu retrotransposition. Importantly, the anti-L1 activity of hA3G was also associated with hA3G oligomerization. These results suggest that the inhibitory activities of hA3G against Alu and L1 retrotransposition might involve a common mechanism.

  6. Modulation of the oligomerization of myelin proteolipid protein by transmembrane helix interaction motifs.

    Science.gov (United States)

    Ng, Derek P; Deber, Charles M

    2010-08-17

    Proteolipid protein (PLP) is a highly hydrophobic 276-residue integral membrane protein that constitutes more than 50% of the total protein in central nervous system myelin. Previous studies have shown that this protein exists in myelin as an oligomer rather than as a monomer, and mutations in PLP that lead to neurological disorders such as Pelizaeus-Merzbacher disease and spastic paraplegia type 2 have been reported to affect its normal oligomerization. Here we employ peptide-based and in vivo approaches to examine the role of the TM domain in the formation of PLP quaternary structure through homo-oligomeric helix-helix interactions. Focusing on the TM4 alpha-helix (sequence (239)FIAAFVGAAATLVSLLTFMIAATY(262)), the site of several disease-causing point mutations that involve putative small residue helix-helix interaction motifs in the TM4 sequence, we used SDS-PAGE, fluorescence resonance energy transfer, size-exclusion chromatography, and TOXCAT assays in an Escherichia coli membrane to show that the PLP TM4 helix readily assembles into varying oligomeric states. In addition, through targeted studies of the PLP TM4 alpha-helix with point mutations that selectively eliminate these small residue motifs via substitution of Gly, Ala, or Ser residues with Ile residues, we describe a potential mechanism through which disease-causing point mutations can lead to aberrant PLP assembly. The overall results suggest that TM segments in misfolded PLP monomers that expose and/or create surface-exposed helix-helix interaction sites that are normally masked may have consequences for disease.

  7. Oligomeric structure of a cathelicidin antimicrobial peptide in dodecylphosphocholine micelle determined by NMR spectroscopy.

    Science.gov (United States)

    Saravanan, Rathi; Bhattacharjya, Surajit

    2011-01-01

    The broad spectrum of antibacterial activities of host defense cationic antimicrobial peptides (AMPs) arises from their ability to perturb membrane integrity of the microbes. The mechanisms are often thought to require assembly of AMPs on the membrane surface to form pores. However, three dimensional structures in the oligomeric form of AMPs in the context of lipid membranes are largely limited. Here, we demonstrate that a 22-residue antimicrobial peptide, termed VK22, derived from fowlicidin-1, a cathelicidin family of AMP from chicken oligomerizes into a predominantly tetrameric state in zwitterionic dodecylphosphocholine (DPC) micelles. An ensemble of NMR structures of VK22 determined in 200mM perdeuterated DPC, from 755 NOE constrains including 19 inter-helical NOEs, had revealed an assembly of four helices arranged in anti-parallel fashion. Hydrogen bonds, C(α)H-O=C types, and van der Waals interactions among the helical sub-units appear to be involved in the stabilization of the quaternary structures. The central region of the barrel shaped tetrameric bundle is non-polar with clusters of aromatic residues, whereas all the cationic residues are positioned at the termini. Paramagnetic spin labeled NMR experiments indicated that the tetrameric structure is embedded into micelles such that the non-polar region located inside the lipid acyl chains. Structure and micelle localization of a monomeric version, obtained from substitution of two Tyr residues with Ala, of the peptide is also compared. The mutated peptide VK22AA has been found be localized at the surface of the micelles. The tetrameric structure of VK22 delineates a small water pore that can be larger in the higher order oligomers. As these results provide structural insights, at atomic resolution, into the oligomeric states of a helical AMP in lipid environment, the structural details may be further utilized for the design of novel self-assembled membrane protein mimics.

  8. Crystal structures from the Plasmodium peroxiredoxins: new insights into oligomerization and product binding

    Directory of Open Access Journals (Sweden)

    Qiu Wei

    2012-03-01

    Full Text Available Abstract Background Plasmodium falciparum is the protozoan parasite primarily responsible for more than one million malarial deaths, annually, and is developing resistance to current therapies. Throughout its lifespan, the parasite is subjected to oxidative attack, so Plasmodium antioxidant defences are essential for its survival and are targets for disease control. Results To further understand the molecular aspects of the Plasmodium redox system, we solved 4 structures of Plasmodium peroxiredoxins (Prx. Our study has confirmed PvTrx-Px1 to be a hydrogen peroxide (H2O2-sensitive peroxiredoxin. We have identified and characterized the novel toroid octameric oligomer of PyTrx-Px1, which may be attributed to the interplay of several factors including: (1 the orientation of the conserved surface/buried arginine of the NNLA(I/LGRS-loop; and (2 the C-terminal tail positioning (also associated with the aforementioned conserved loop which facilitates the intermolecular hydrogen bond between dimers (in an A-C fashion. In addition, a notable feature of the disulfide bonds in some of the Prx crystal structures is discussed. Finally, insight into the latter stages of the peroxiredoxin reaction coordinate is gained. Our structure of PyPrx6 is not only in the sulfinic acid (RSO2H form, but it is also with glycerol bound in a way (not previously observed indicative of product binding. Conclusions The structural characterization of Plasmodium peroxiredoxins provided herein provides insight into their oligomerization and product binding which may facilitate the targeting of these antioxidant defences. Although the structural basis for the octameric oligomerization is further understood, the results yield more questions about the biological implications of the peroxiredoxin oligomerization, as multiple toroid configurations are now known. The crystal structure depicting the product bound active site gives insight into the overoxidation of the active site and

  9. Self-formed waterfall plunge pools in homogeneous rock

    Science.gov (United States)

    Scheingross, Joel S.; Lo, Daniel Y.; Lamb, Michael P.

    2017-01-01

    Waterfalls are ubiquitous, and their upstream propagation can set the pace of landscape evolution, yet no experimental studies have examined waterfall plunge pool erosion in homogeneous rock. We performed laboratory experiments, using synthetic foam as a bedrock simulant, to produce self-formed waterfall plunge pools via particle impact abrasion. Plunge pool vertical incision exceeded lateral erosion by approximately tenfold until pools deepened to the point that the supplied sediment could not be evacuated and deposition armored the pool bedrock floor. Lateral erosion of plunge pool sidewalls continued after sediment deposition, but primarily at the downstream pool wall, which might lead to undermining of the plunge pool lip, sediment evacuation, and continued vertical pool floor incision in natural streams. Undercutting of the upstream pool wall was absent, and our results suggest that vertical drilling of successive plunge pools is a more efficient waterfall retreat mechanism than the classic model of headwall undercutting and collapse in homogeneous rock.

  10. Humanitarian engineering in the engineering curriculum

    Science.gov (United States)

    Vandersteen, Jonathan Daniel James

    There are many opportunities to use engineering skills to improve the conditions for marginalized communities, but our current engineering education praxis does not instruct on how engineering can be a force for human development. In a time of great inequality and exploitation, the desire to work with the impoverished is prevalent, and it has been proposed to adjust the engineering curriculum to include a larger focus on human needs. This proposed curriculum philosophy is called humanitarian engineering. Professional engineers have played an important role in the modern history of power, wealth, economic development, war, and industrialization; they have also contributed to infrastructure, sanitation, and energy sources necessary to meet human need. Engineers are currently at an important point in time when they must look back on their history in order to be more clear about how to move forward. The changing role of the engineer in history puts into context the call for a more balanced, community-centred engineering curriculum. Qualitative, phenomenographic research was conducted in order to understand the need, opportunity, benefits, and limitations of a proposed humanitarian engineering curriculum. The potential role of the engineer in marginalized communities and details regarding what a humanitarian engineering program could look like were also investigated. Thirty-two semi-structured research interviews were conducted in Canada and Ghana in order to collect a pool of understanding before a phenomenographic analysis resulted in five distinct outcome spaces. The data suggests that an effective curriculum design will include teaching technical skills in conjunction with instructing about issues of social justice, social location, cultural awareness, root causes of marginalization, a broader understanding of technology, and unlearning many elements about the role of the engineer and the dominant economic/political ideology. Cross-cultural engineering development

  11. Isolation and identification of oligomeric procyanidins from Crataegus leaves and flowers.

    Science.gov (United States)

    Svedström, Ulla; Vuorela, Heikki; Kostiainen, Risto; Tuominen, Jari; Kokkonen, Juha; Rauha, Jussi-Pekka; Laakso, Into; Hiltunen, Raimo

    2002-08-01

    Oligomeric procyanidins were isolated from the leaves and flowers of hawthorn (Crataegus laevigata). A trimer, epicatechin-(4 beta-->8)-epicatechin-(4 beta-->6)-epicatechin, and a pentamer consisting of (-)-epicatechin units linked through C-4 beta/C-8 bonds have been isolated from hawthorn for the first time, in addition to known procyanidins including dimers B-2, B-4 and B-5, trimers C-1 and epicatechin-(4 beta-->6)-epicatechin-(4 beta-->8)-epicatechin, and tetramer D-1. A fraction containing a hexamer was also found.

  12. Thermo-mechanical characterization of a monochlorophenyl, hepta isobutyl polyhedral oligomeric silsesquioxane/polystyrene composite

    Energy Technology Data Exchange (ETDEWEB)

    Blanco, Ignazio, E-mail: iblanco@dii.unict.it; Bottino, Francesco A., E-mail: iblanco@dii.unict.it; Cicala, Gianluca, E-mail: iblanco@dii.unict.it; Cozzo, Giulia, E-mail: iblanco@dii.unict.it; Latteri, Alberta, E-mail: iblanco@dii.unict.it; Recca, Antonino, E-mail: iblanco@dii.unict.it [Department of Industrial Engineering, University of Catania, Viale A. Doria 6, 95125 Catania (Italy)

    2014-05-15

    The thermal and mechanical properties of a monochlorophenyl, hepta isobutyl Polyhedral Oligomeric Silsesquioxane/Polystyrene (ph,hib-POSS/PS) composite were studied and compared with those of pristine polymer. ph,hib-POSS/PS system was prepared by solubilization and precipitation of Polystyrene (PS) in the presence of POSS. Scanning Electron Microscopy (SEM) was performed to check the distribution of the filler in the polymer matrix. Dynamic Mechanical Analysis (DMA) was carried out to measure viscoelastic properties of solid samples. Degradations were carried out into a thermobalance and the obtained thermogravimetric (TG) and differential thermogravimetric (DTG) curves were discussed and interpreted.

  13. Esters of oligo-(glycerol carbonate-glycerol): New biobased oligomeric surfactants.

    Science.gov (United States)

    Holmiere, Sébastien; Valentin, Romain; Maréchal, Philippe; Mouloungui, Zéphirin

    2017-02-01

    Glycerol carbonate is one of the most potentially multifunction glycerol-derived compounds. Glycerol is an important by-product of the oleochemical industry. The oligomerization of glycerol carbonate, assisted by the glycerol, results in the production of polyhydroxylated oligomers rich in linear carbonate groups. The polar moieties of these oligomers (Mwesters of sorbitan polyethoxylates. The self-assembling properties of oligocarbonate esters were highlighted by their ability to stabilize inverse and multiple emulsions. The oligo-(glycerol carbonate-glycerol ether) with relatively low molecular weights showed properties of relatively high-molecular weight molecules, and constitute a viable "green" alternative to ethoxylated surfactants. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. [Theory of V.A. dogiel on polymerization and oligomerization as a general integration concept].

    Science.gov (United States)

    Makmaev, Iu V

    2010-01-01

    The theory of V.A. Dogiel on the significance of polymerization and ligomerization processes in the evolution of Protozoa and Metazoa is compared with the paper of I.I. Schmalhauisen (1972) on factors and steps of aromorph evolution. Dogiel's theory is considered as a general integration conception. Four steps are distinguished in the evolution of biological systems: (1) formation of morphofunctional system by units of the lower structural level, (2) polymerization of morphofunctional units of a system, (3) oligomerization of morphofunctional units of system by means of their reduction, uniting, or differentiation, (4) integration and stabilization of a system owing to development of morphofunctional connections between its parts.

  15. Multi-PAS domain-mediated protein oligomerization of PpsR from Rhodobacter sphaeroides

    Energy Technology Data Exchange (ETDEWEB)

    Heintz, Udo; Meinhart, Anton; Winkler, Andreas, E-mail: andreas.winkler@mpimf-heidelberg.mpg.de [Max Planck Institute for Medical Research, Heidelberg (Germany)

    2014-03-01

    Crystal structures of two truncated variants of the transcription factor PpsR from R. sphaeroides are presented that enabled the phasing of a triple PAS domain construct. Together, these structures reveal the importance of α-helical PAS extensions for multi-PAS domain-mediated protein oligomerization and function. Per–ARNT–Sim (PAS) domains are essential modules of many multi-domain signalling proteins that mediate protein interaction and/or sense environmental stimuli. Frequently, multiple PAS domains are present within single polypeptide chains, where their interplay is required for protein function. Although many isolated PAS domain structures have been reported over the last decades, only a few structures of multi-PAS proteins are known. Therefore, the molecular mechanism of multi-PAS domain-mediated protein oligomerization and function is poorly understood. The transcription factor PpsR from Rhodobacter sphaeroides is such a multi-PAS domain protein that, in addition to its three PAS domains, contains a glutamine-rich linker and a C-terminal helix–turn–helix DNA-binding motif. Here, crystal structures of two N-terminally and C-terminally truncated PpsR variants that comprise a single (PpsR{sub Q-PAS1}) and two (PpsR{sub N-Q-PAS1}) PAS domains, respectively, are presented and the multi-step strategy required for the phasing of a triple PAS domain construct (PpsR{sub ΔHTH}) is illustrated. While parts of the biologically relevant dimerization interface can already be observed in the two shorter constructs, the PpsR{sub ΔHTH} structure reveals how three PAS domains enable the formation of multiple oligomeric states (dimer, tetramer and octamer), highlighting that not only the PAS cores but also their α-helical extensions are essential for protein oligomerization. The results demonstrate that the long helical glutamine-rich linker of PpsR results from a direct fusion of the N-cap of the PAS1 domain with the C-terminal extension of the N-domain that

  16. Thermo-mechanical characterization of a monochlorophenyl, hepta isobutyl polyhedral oligomeric silsesquioxane/polystyrene composite

    Science.gov (United States)

    Blanco, Ignazio; Bottino, Francesco A.; Cicala, Gianluca; Cozzo, Giulia; Latteri, Alberta; Recca, Antonino

    2014-05-01

    The thermal and mechanical properties of a monochlorophenyl, hepta isobutyl Polyhedral Oligomeric Silsesquioxane/Polystyrene (ph,hib-POSS/PS) composite were studied and compared with those of pristine polymer. ph,hib-POSS/PS system was prepared by solubilization and precipitation of Polystyrene (PS) in the presence of POSS. Scanning Electron Microscopy (SEM) was performed to check the distribution of the filler in the polymer matrix. Dynamic Mechanical Analysis (DMA) was carried out to measure viscoelastic properties of solid samples. Degradations were carried out into a thermobalance and the obtained thermogravimetric (TG) and differential thermogravimetric (DTG) curves were discussed and interpreted.

  17. Engineering Encounters: Engineering Adaptations

    Science.gov (United States)

    Gatling, Anne; Vaughn, Meredith Houle

    2015-01-01

    Engineering is not a subject that has historically been taught in elementary schools, but with the emphasis on engineering in the "Next Generation Science Standards," curricula are being developed to explicitly teach engineering content and design. However, many of the scientific investigations already conducted with students have…

  18. Engineering Encounters: Engineering Adaptations

    Science.gov (United States)

    Gatling, Anne; Vaughn, Meredith Houle

    2015-01-01

    Engineering is not a subject that has historically been taught in elementary schools, but with the emphasis on engineering in the "Next Generation Science Standards," curricula are being developed to explicitly teach engineering content and design. However, many of the scientific investigations already conducted with students have…

  19. Reserve Growth of Alberta Oil Pools

    Science.gov (United States)

    Verma, Mahendra K.; Cook, Troy

    2008-01-01

    This Open-File Report is based on a presentation delivered at the Fourth U.S. Geological Survey Workshop on Reserve Growth on March 10-11, 2008. It summarizes the results of a study of reserve growth of oil pools in Alberta Province, Canada. The study is part of a larger effort involving similar studies of fields in other important petroleum provinces around the world, with the overall objective of gaining a better understanding of reserve growth in fields with different geologic/reservoir parameters and different operating environments. The goals of the study were to: 1. Evaluate historical oil reserve data and assess reserve growth. 2. Develop reserve growth models/functions to help forecast hydrocarbon volumes. 3. Study reserve growth sensitivity to various parameters ? for example, pool size, porosity, oil gravity, and lithology. 4. Compare reserve growth in oil pools/fields of Alberta provinces with those from other large petroleum provinces.

  20. SNP calling by sequencing pooled samples

    Directory of Open Access Journals (Sweden)

    Raineri Emanuele

    2012-09-01

    Full Text Available Abstract Background Performing high throughput sequencing on samples pooled from different individuals is a strategy to characterize genetic variability at a small fraction of the cost required for individual sequencing. In certain circumstances some variability estimators have even lower variance than those obtained with individual sequencing. SNP calling and estimating the frequency of the minor allele from pooled samples, though, is a subtle exercise for at least three reasons. First, sequencing errors may have a much larger relevance than in individual SNP calling: while their impact in individual sequencing can be reduced by setting a restriction on a minimum number of reads per allele, this would have a strong and undesired effect in pools because it is unlikely that alleles at low frequency in the pool will be read many times. Second, the prior allele frequency for heterozygous sites in individuals is usually 0.5 (assuming one is not analyzing sequences coming from, e.g. cancer tissues, but this is not true in pools: in fact, under the standard neutral model, singletons (i.e. alleles of minimum frequency are the most common class of variants because P(f ∝ 1/f and they occur more often as the sample size increases. Third, an allele appearing only once in the reads from a pool does not necessarily correspond to a singleton in the set of individuals making up the pool, and vice versa, there can be more than one read – or, more likely, none – from a true singleton. Results To improve upon existing theory and software packages, we have developed a Bayesian approach for minor allele frequency (MAF computation and SNP calling in pools (and implemented it in a program called snape: the approach takes into account sequencing errors and allows users to choose different priors. We also set up a pipeline which can simulate the coalescence process giving rise to the SNPs, the pooling procedure and the sequencing. We used it to compare the

  1. Mechanisms for convection triggering by cold pools

    CERN Document Server

    Torri, Giuseppe; Tian, Yang

    2015-01-01

    Cold pools are fundamental ingredients of deep convection. They contribute to organizing the sub-cloud layer and are considered key elements in triggering convective cells. It was long known that this could happen mechanically, through lifting by the cold pools' fronts. More recently, it has been suggested that convection could also be triggered thermodynamically, by accumulation of moisture around the edges of cold pools. A method based on Lagrangian tracking is here proposed to disentangle the signatures of both forcings and quantify their importance in a given environment. Results from a simulation of radiative-convective equilibrium over the ocean show that parcels reach their level of free convection through a combination of both forcings, each being dominant at different stages of the ascent. Mechanical forcing is an important player in lifting parcels from the surface, whereas thermodynamic forcing reduces the inhibition encountered by parcels before they reach their level of free convection.

  2. Disassembly intermediates of RbsD protein remain oligomeric despite the loss of an intact secondary structure

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    Many proteins exist as homo-oligomers in living organisms wherein the change of oligomeric status apparently serves as an effective means for modulating their biological activities. We have previously reported that the homo-decameric RbsD from Escherichia coli undergoes stepwise disassembly and non-stepwise reassembly. Here the structural status of the urea-induced RbsD disassembly intermediates was examined, mainly using urea-containing polyacrylamide gel electrophoresis and chemical cross-linking. Such intermediates were found to remain oligomeric while losing their intact secondary structures. Such disassembly intermediates were able to effectively refold when the concentration of the urea denaturant was reduced to a lower level, or to refold/reassemble into the native decamers when urea was completely removed, as detected by non-denaturing polyacrylamide gel electrophoresis. These novel observations strongly suggest that the assembly of oligomeric proteins may occur before the completion of subunit folding.

  3. Structures and formation mechanisms of aquo/hydroxo oligomeric beryllium in aqueous solution: a density functional theory study.

    Science.gov (United States)

    Jin, Xiaoyan; Liao, Rongbao; Wu, Hai; Huang, Zhengjie; Zhang, Hong

    2015-09-01

    The structures and formation mechanisms of a wide variety of aquo/hydroxo oligomeric beryllium clusters were investigated using density functional theory. The structural parameters of beryllium clusters were found to vary regularly with the stepwise substitution of bound water molecules in the inner coordination sphere by hydroxyl groups. According to the Gibbs free energies deduced from SMD solvation model computations, unhydrolyzed oligomeric beryllium species are the most favorable products of polymerization, independent of the degrees of hydrolysis of the reactants. Simulation of the formation processes of oligomeric beryllium showed that polymerization, in essence, involves the nucleophilic attack of a terminal hydroxyl group in one BeO4 tetrahedron on the beryllium center in another BeO4 tetrahedron, leading to the bridging of two BeO4 tetrahedrons by a hydroxyl group.

  4. Gene pool conservation and tree improvement in Serbia

    Directory of Open Access Journals (Sweden)

    Isajev Vasilije

    2009-01-01

    Full Text Available This paper presents the concepts applied in the gene pool conservation and tree improvement in Serbia. Gene pool conservation of tree species in Serbia includes a series of activities aiming at the sustainability and protection of genetic and species variability. This implies the investigation of genetic resources and their identification through the research of the genetic structure and the breeding system of individual species. Paper also includes the study of intra- and inter-population variability in experiments - provenance tests, progeny tests, half- and full-sib lines, etc. The increased use of the genetic potential in tree improvement in Serbia should be intensified by the following activities: improvement of production of normal forest seed, application of the concept of new selections directed primarily to the improvement of only one character, because in that case the result would be certain, establishment and management of seed orchards as specialized plantations for long-term production of genetically good-quality forest seeds, and the shortening of the improvement process by introducing new techniques and methods (molecular markers, somaclonal variation, genetic engineering, protoplast fusion, micropropagation, etc..

  5. Effects of indoor swimming pools on the nasal cytology of pool workers.

    Science.gov (United States)

    Erkul, E; Yaz, A; Cıngı, C; İnançli, H M; San, T; Bal, C

    2014-05-01

    We aimed to evaluate the relationship between swimming pool pollutants and allergic rhinitis in swimming pool workers. Twenty-seven indoor pool workers (group 1) and 49 control subjects (group 2) were enrolled in the study. A skin prick test was performed and a nasal smear was obtained from each subject to evaluate rhinitis. When the groups were compared in terms of epithelial cells, group 1 had significantly more epithelial cells than group 2. When the groups were compared with regard to eosinophils, group 1 had significantly more eosinophils than group 2. The skin prick test results for both groups were not significantly different. Indoor pool workers showed severe symptoms of rhinitis and eosinophilic nasal cytology, likely due to chlorine. Nasal cytology is an easy-to-administer diagnostic test and can be used to follow up rhinitis in indoor pool workers, along with nasal endoscopy, a detailed clinical history and a skin prick test.

  6. Topography Battles Surface Texture: An Experimental Study of Pool-riffle Formation

    Science.gov (United States)

    Chartrand, S. M.; Hassan, M. A.; Jellinek, M.

    2016-12-01

    Pool-riffles are perhaps the most common streambed shape found in streams and rivers, and not surprisingly, they are essential to salmon ecology, and are a central focus of many restoration actions. Yet, when an applied earth scientist or engineer is faced with developing a pool-riffle design, there is a lack of clear and rigorously developed design guidelines. Given the volumes of money spent annually within the restoration industry, this is a real problem. Recognition of this problem is growing, however, and an increasing level of attention has been directed to questions of pool-riffle formation in the past decade. At this point and given certain landscape characteristics, it is well established that streamwise gradients in channel width are associated with pool-riffles. Specifically, pools are associated with negative gradients in width, and riffles with positive gradients. Importantly, these associations have now been documented from field-derived data, as well as via experimental and numerical investigations. There is much to build from the present knowledge base, and central to this are questions related to (a) how pool-riffles evolve during the formative process, (b) what are the basic set of ingredients necessary for pool-riffle formation within systems characterized by relatively non-erodible channel margins, and (c) do pool-riffles persist, once formed, under a broad range of forcing conditions? We have completed four physical experiments examining the process and evolution of pool-riffle formation under a large range of upstream boundary, as well as physical channel conditions. We will report on two of the completed experiments. Our work will highlight two new non-dimensional channel evolution numbers, derived to help describe and characterize bedform development, as well as response to perturbations from near-equilibrium conditions. The channel evolution numbers lay the foundation for development of a new regime diagram, which quantifies the basic

  7. 47 CFR 90.35 - Industrial/Business Pool.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 5 2010-10-01 2010-10-01 false Industrial/Business Pool. 90.35 Section 90.35... MOBILE RADIO SERVICES Industrial/Business Radio Pool § 90.35 Industrial/Business Pool. (a) Eligibility... Industrial/Business Pool to provide commercial mobile radio service as defined in part 20 of this chapter...

  8. 48 CFR 232.470 - Advance payment pool.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 3 2010-10-01 2010-10-01 false Advance payment pool. 232... Items 232.470 Advance payment pool. (a) An advance payment pool agreement— (1) Is a means of financing... addition to any other advance payment pool agreement at a single contractor location when it is...

  9. 47 CFR 90.20 - Public Safety Pool.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 5 2010-10-01 2010-10-01 false Public Safety Pool. 90.20 Section 90.20... MOBILE RADIO SERVICES Public Safety Radio Pool § 90.20 Public Safety Pool. (a) Eligibility. The following are eligible to hold authorizations in the Public Safety Pool. (1) Any territory, possession,...

  10. Adrian Poole. Shakespeare and the Victorians

    Directory of Open Access Journals (Sweden)

    Béatrice LAURENT

    2006-06-01

    Full Text Available Dr Adrian Poole of Trinity College, Cambridge offers in this new study a comprehensive panorama of Shakespeare’s influence on the Victorians. Dr Poole investigates the interpretation and appropriation of the plays and, to a lesser extent, of the poems by Shakespeare with a view to demonstrate that they provided a major source of inspiration to Victorian actors, novelists and visual artists.The point of view is that of a distinguished Victorianist, and the reader is supposed to have a sound kn...

  11. Zooplankton at deep Red Sea brine pools

    KAUST Repository

    Kaartvedt, Stein

    2016-03-02

    The deep-sea anoxic brines of the Red Sea comprise unique, complex and extreme habitats. These environments are too harsh for metazoans, while the brine–seawater interface harbors dense microbial populations. We investigated the adjacent pelagic fauna at two brine pools using net tows, video records from a remotely operated vehicle and submerged echosounders. Waters just above the brine pool of Atlantis II Deep (2000 m depth) appeared depleted of macrofauna. In contrast, the fauna appeared to be enriched at the Kebrit Deep brine–seawater interface (1466 m).

  12. The new pooled cohort equations risk calculator

    DEFF Research Database (Denmark)

    Preiss, David; Kristensen, Søren L

    2015-01-01

    total cardiovascular risk score. During development of joint guidelines released in 2013 by the American College of Cardiology (ACC) and American Heart Association (AHA), the decision was taken to develop a new risk score. This resulted in the ACC/AHA Pooled Cohort Equations Risk Calculator. This risk...... disease and any measure of social deprivation. An early criticism of the Pooled Cohort Equations Risk Calculator has been its alleged overestimation of ASCVD risk which, if confirmed in the general population, is likely to result in statin therapy being prescribed to many individuals at lower risk than...

  13. Censored Exploration and the Dark Pool Problem

    CERN Document Server

    Ganchev, Kuzman; Nevmyvaka, Yuriy; Vaughan, Jennifer Wortman

    2012-01-01

    We introduce and analyze a natural algorithm for multi-venue exploration from censored data, which is motivated by the Dark Pool Problem of modern quantitative finance. We prove that our algorithm converges in polynomial time to a near-optimal allocation policy; prior results for similar problems in stochastic inventory control guaranteed only asymptotic convergence and examined variants in which each venue could be treated independently. Our analysis bears a strong resemblance to that of efficient exploration/ exploitation schemes in the reinforcement learning literature. We describe an extensive experimental evaluation of our algorithm on the Dark Pool Problem using real trading data.

  14. [Pool exercise therapy of rheumatoid arthritis].

    Science.gov (United States)

    Bartels, E M; Lund, H; Danneskiold-Samsøe, B

    2001-10-01

    Aquatic therapy is a subgroup of balneotherapy and consists of exercises in a hot water pool. It uses the physical properties of water to achieve better mobility for patients whose pain, lack of muscle strength, and joint deformities are inhibiting factors when exercising on land. Pool therapy shows positive effects as part of the treatment of rheumatoid arthritis patients, but too few studies with an acceptable design and a well-defined patient group have been carried out. The documentation available on aquatic therapy indicates that more large clinical, controlled, and randomised studies must be conducted.

  15. The N-terminus of TDP-43 promotes its oligomerization and enhances DNA binding affinity

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Chung-ke [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Wu, Tzong-Huah [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China); Chemical Biology and Molecular Biophysics Program, Taiwan International Graduate Program, Institute of Biochemistry, Academia Sinica, Taipei 115, Taiwan (China); Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 300, Taiwan (China); Wu, Chu-Ya [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China); Graduate Institute of Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan (China); Chiang, Ming-hui; Toh, Elsie Khai-Woon [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Hsu, Yin-Chih; Lin, Ku-Feng [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China); Liao, Yu-heng [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Huang, Tai-huang, E-mail: bmthh@gate.sinica.edu.tw [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Department of Physics, National Taiwan Normal University, Taipei 106, Taiwan (China); Huang, Joseph Jen-Tse, E-mail: jthuang@chem.sinica.edu.tw [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China)

    2012-08-24

    Highlights: Black-Right-Pointing-Pointer The N-terminus of TDP-43 contains an independently folded structural domain (NTD). Black-Right-Pointing-Pointer The structural domains of TDP-43 are arranged in a beads-on-a-string fashion. Black-Right-Pointing-Pointer The NTD promotes TDP-43 oligomerization in a concentration-dependent manner. Black-Right-Pointing-Pointer The NTD may assist nucleic acid-binding activity of TDP-43. -- Abstract: TDP-43 is a DNA/RNA-binding protein associated with different neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD-U). Here, the structural and physical properties of the N-terminus on TDP-43 have been carefully characterized through a combination of nuclear magnetic resonance (NMR), circular dichroism (CD) and fluorescence anisotropy studies. We demonstrate for the first time the importance of the N-terminus in promoting TDP-43 oligomerization and enhancing its DNA-binding affinity. An unidentified structural domain in the N-terminus is also disclosed. Our findings provide insights into the N-terminal domain function of TDP-43.

  16. Zinc ion conducting polymer electrolytes based on oligomeric polyether/PVDF-HFP blends

    Science.gov (United States)

    Ye, Hui; Xu, Jun John

    Here we report novel zinc ion conducting polymer electrolytes based on oligomeric polyether/PVDF-HFP blends with or without the incorporation of a small amount of organic carbonates. Their thermal properties, ionic conductivity and electrochemical properties are characterized and the effect of different Zn salts and incorporation of a small amount of organic carbonates are investigated. These polymer electrolyte membranes exhibit essentially no or very low volatility, high thermal stability, high ionic conductivity, wide electrochemical stability window, acceptable interfacial resistance with zinc, and the capability for reversible Zn plating/stripping. Particularly promising are electrolyte systems based on the combination of low lattice energy zinc imide salt and a special co-solvent of oligomeric poly(ethylene glycol) dimethyl ether (PEGDME) mixed with a small amount of ethylene carbonate (EC), dimensionally stabilized with PVDF-HFP. Such novel polymer electrolyte membranes could lead to the development of new kinds of electrochemical energy storage devices based on zinc electrochemistry, including solid-state, thin-film rechargeable zinc/air cells envisaged.

  17. Zinc ion conducting polymer electrolytes based on oligomeric polyether/PVDF-HFP blends

    Energy Technology Data Exchange (ETDEWEB)

    Ye, Hui; Xu, Jun John [Department of Materials Science and Engineering, Rutgers, the State University of New Jersey, Piscataway, NJ 08854 (United States)

    2007-03-20

    Here we report novel zinc ion conducting polymer electrolytes based on oligomeric polyether/PVDF-HFP blends with or without the incorporation of a small amount of organic carbonates. Their thermal properties, ionic conductivity and electrochemical properties are characterized and the effect of different Zn salts and incorporation of a small amount of organic carbonates are investigated. These polymer electrolyte membranes exhibit essentially no or very low volatility, high thermal stability, high ionic conductivity, wide electrochemical stability window, acceptable interfacial resistance with zinc, and the capability for reversible Zn plating/stripping. Particularly promising are electrolyte systems based on the combination of low lattice energy zinc imide salt and a special co-solvent of oligomeric poly(ethylene glycol) dimethyl ether (PEGDME) mixed with a small amount of ethylene carbonate (EC), dimensionally stabilized with PVDF-HFP. Such novel polymer electrolyte membranes could lead to the development of new kinds of electrochemical energy storage devices based on zinc electrochemistry, including solid-state, thin-film rechargeable zinc/air cells envisaged. (author)

  18. Oligomerization of SCFTIR1 Is Essential for Aux/IAA Degradation and Auxin Signaling in Arabidopsis.

    Science.gov (United States)

    Dezfulian, Mohammad H; Jalili, Espanta; Roberto, Don Karl A; Moss, Britney L; Khoo, Kerry; Nemhauser, Jennifer L; Crosby, William L

    2016-09-01

    The phytohormone auxin is a key regulator of plant growth and development. Molecular studies in Arabidopsis have shown that auxin perception and signaling is mediated via TIR1/AFB-Aux/IAA co-receptors that assemble as part of the SCFTIR1/AFB E3 ubiquitin-ligase complex and direct the auxin-regulated degradation of Aux/IAA transcriptional repressors. Despite the importance of auxin signaling, little is known about the functional regulation of the TIR1/AFB receptor family. Here we show that TIR1 can oligomerize in planta via a set of spatially clustered amino acid residues. While none of the residues identified reside in the interaction interface of the TIR1-Aux/IAA degron, they nonetheless regulate the binding of TIR1 to Aux/IAA substrate proteins and their subsequent degradation in vivo as an essential aspect of auxin signaling. We propose oligomerization of TIR1 as a novel regulatory mechanism in the regulation of auxin-mediated plant patterning and development.

  19. Glycation of Wild-Type Apomyoglobin Induces Formation of Highly Cytotoxic Oligomeric Species.

    Science.gov (United States)

    Iannuzzi, Clara; Carafa, Vincenzo; Altucci, Lucia; Irace, Gaetano; Borriello, Margherita; Vinciguerra, Roberto; Sirangelo, Ivana

    2015-11-01

    Protein glycation is a non-enzymatic, irreversible modification of protein amino groups by reactive carbonyl species leading to the formation of advanced glycation end products (AGEs). Several proteins implicated in neurodegenerative diseases have been found to be glycated in vivo and the extent of glycation is related to the pathologies of the patients. Although it is now accepted that there is a direct correlation between AGEs formation and the development of neurodegenerative diseases related to protein misfolding and amyloid aggregation, several questions still remain unanswered: whether glycation is the triggering event or just an additional factor acting on the aggregation pathway. We have recently shown that glycation of the amyloidogenic W7FW14F apomyoglobin mutant significantly accelerates the amyloid fibrils formation providing evidence that glycation actively participates to the process. In the present study, to test if glycation can be considered also a triggering factor in amyloidosis, we evaluated the ability of different glycation agents to induce amyloid aggregation in the soluble wild-type apomyoglobin. Our results show that glycation covalently modifies apomyoglobin and induces conformational changes that lead to the formation of oligomeric species that are not implicated in amyloid aggregation. Thus, AGEs formation does not trigger amyloid aggregation in the wild-type apomyoglobin but only induce the formation of soluble oligomeric species able to affect cell viability. The molecular bases of cell toxicity induced by AGEs formed upon glycation of wild-type apomyoglobin have been also investigated.

  20. Protein Cross-Linking and Oligomerization through Dityrosine Formation upon Exposure to Ozone.

    Science.gov (United States)

    Kampf, Christopher J; Liu, Fobang; Reinmuth-Selzle, Kathrin; Berkemeier, Thomas; Meusel, Hannah; Shiraiwa, Manabu; Pöschl, Ulrich

    2015-09-15

    Air pollution is a potential driver for the increasing prevalence of allergic disease, and post-translational modification by air pollutants can enhance the allergenic potential of proteins. Here, the kinetics and mechanism of protein oligomerization upon ozone (O3) exposure were studied in coated-wall flow tube experiments at environmentally relevant O3 concentrations, relative humidities and protein phase states (amorphous solid, semisolid, and liquid). We observed the formation of protein dimers, trimers, and higher oligomers, and attribute the cross-linking to the formation of covalent intermolecular dityrosine species. The oligomerization proceeds fast on the surface of protein films. In the bulk material, reaction rates are limited by diffusion depending on phase state and humidity. From the experimental data, we derive a chemical mechanism and rate equations for a kinetic multilayer model of surface and bulk reaction enabling the prediction of oligomer formation. Increasing levels of tropospheric O3 in the Anthropocene may promote the formation of protein oligomers with enhanced allergenicity and may thus contribute to the increasing prevalence of allergies.

  1. Quercetin Inhibits Inflammasome Activation by Interfering with ASC Oligomerization and Prevents Interleukin-1 Mediated Mouse Vasculitis

    Science.gov (United States)

    Domiciano, Talita P.; Wakita, Daiko; Jones, Heather D.; Crother, Timothy R.; Verri, Waldiceu A.; Arditi, Moshe; Shimada, Kenichi

    2017-01-01

    Interleukin-1β (IL-1β) is a highly inflammatory cytokine that significantly contributes to both acute and chronic inflammatory diseases. The secretion of IL-1β requires a unique protease, caspase-1, which is activated by various protein platforms called inflammasomes. Data suggests a key role for mitochondrial reactive oxygen species for inflammasome activation. Flavonoids constitute a group of naturally occurring polyphenolic molecules with many biological activities, including antioxidant effects. In this study, we investigated the effect of three flavonoids, quercetin (QUC), naringenin, and silymarim on inflammasome activation. We found that QUC inhibits IL-1β secretion by both the NLRP3 and AIM2 inflammasome in a dose dependent manner, but not the NLRC4 inflammasome. QUC inhibition of the inflammasome was still observed in Atg16l1 knockout macrophages, indicating that QUC’s effect was autophagy independent. Since QUC inhibited both NLRP3 and AIM2 inflammasomes but not NLRC4, we assessed ASC speck formation. QUC reduced ASC speck formation and ASC oligomerization compared with controls. Additionally, QUC inhibited IL-1β in Cryopyrin-Associated Periodic Syndromes (CAPS) macrophages, where NLRP3 inflammasome is constitutively activated. In conclusion, QUC inhibits both the NLRP3 and AIM2 inflammasome by preventing ASC oligomerization and may be a potential therapeutic candidate for Kawasaki disease vasculitis and other IL-1 mediated inflammatory diseases. PMID:28148962

  2. Characterization of Lassa virus glycoprotein oligomerization and influence of cholesterol on virus replication.

    Science.gov (United States)

    Schlie, Katrin; Maisa, Anna; Lennartz, Frank; Ströher, Ute; Garten, Wolfgang; Strecker, Thomas

    2010-01-01

    Mature glycoprotein spikes are inserted in the Lassa virus envelope and consist of the distal subunit GP-1, the transmembrane-spanning subunit GP-2, and the signal peptide, which originate from the precursor glycoprotein pre-GP-C by proteolytic processing. In this study, we analyzed the oligomeric structure of the viral surface glycoprotein. Chemical cross-linking studies of mature glycoprotein spikes from purified virus revealed the formation of trimers. Interestingly, sucrose density gradient analysis of cellularly expressed glycoprotein showed that in contrast to trimeric mature glycoprotein complexes, the noncleaved glycoprotein forms monomers and oligomers spanning a wide size range, indicating that maturation cleavage of GP by the cellular subtilase SKI-1/S1P is critical for formation of the correct oligomeric state. To shed light on a potential relation between cholesterol and GP trimer stability, we performed cholesterol depletion experiments. Although depletion of cholesterol had no effect on trimerization of the glycoprotein spike complex, our studies revealed that the cholesterol content of the viral envelope is important for the infectivity of Lassa virus. Analyses of the distribution of viral proteins in cholesterol-rich detergent-resistant membrane areas showed that Lassa virus buds from membrane areas other than those responsible for impaired infectivity due to cholesterol depletion of lipid rafts. Thus, derivation of the viral envelope from cholesterol-rich membrane areas is not a prerequisite for the impact of cholesterol on virus infectivity.

  3. Multicoil2: predicting coiled coils and their oligomerization states from sequence in the twilight zone.

    Directory of Open Access Journals (Sweden)

    Jason Trigg

    Full Text Available The alpha-helical coiled coil can adopt a variety of topologies, among the most common of which are parallel and antiparallel dimers and trimers. We present Multicoil2, an algorithm that predicts both the location and oligomerization state (two versus three helices of coiled coils in protein sequences. Multicoil2 combines the pairwise correlations of the previous Multicoil method with the flexibility of Hidden Markov Models (HMMs in a Markov Random Field (MRF. The resulting algorithm integrates sequence features, including pairwise interactions, through multinomial logistic regression to devise an optimized scoring function for distinguishing dimer, trimer and non-coiled-coil oligomerization states; this scoring function is used to produce Markov Random Field potentials that incorporate pairwise correlations localized in sequence. Multicoil2 significantly improves both coiled-coil detection and dimer versus trimer state prediction over the original Multicoil algorithm retrained on a newly-constructed database of coiled-coil sequences. The new database, comprised of 2,105 sequences containing 124,088 residues, includes reliable structural annotations based on experimental data in the literature. Notably, the enhanced performance of Multicoil2 is evident when tested in stringent leave-family-out cross-validation on the new database, reflecting expected performance on challenging new prediction targets that have minimal sequence similarity to known coiled-coil families. The Multicoil2 program and training database are available for download from http://multicoil2.csail.mit.edu.

  4. Integration and oligomerization of Bax protein in lipid bilayers characterized by single molecule fluorescence study.

    Science.gov (United States)

    Luo, Lu; Yang, Jun; Liu, Dongxiang

    2014-11-14

    Bax is a pro-apoptotic Bcl-2 family protein. The activated Bax translocates to mitochondria, where it forms pore and permeabilizes the mitochondrial outer membrane. This process requires the BH3-only activator protein (i.e. tBid) and can be inhibited by anti-apoptotic Bcl-2 family proteins such as Bcl-xL. Here by using single molecule fluorescence techniques, we studied the integration and oligomerization of Bax in lipid bilayers. Our study revealed that Bax can bind to lipid membrane spontaneously in the absence of tBid. The Bax pore formation undergoes at least two steps: pre-pore formation and membrane insertion. The activated Bax triggered by tBid or BH3 domain peptide integrates on bilayers and tends to form tetramers, which are termed as pre-pore. Subsequent insertion of the pre-pore into membrane is highly dependent on the composition of cardiolipin in lipid bilayers. Bcl-xL can translocate Bax from membrane to solution and inhibit the pore formation. The study of Bax integration and oligomerization at the single molecule level provides new evidences that may help elucidate the pore formation of Bax and its regulatory mechanism in apoptosis. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  5. Quercetin Inhibits Inflammasome Activation by Interfering with ASC Oligomerization and Prevents Interleukin-1 Mediated Mouse Vasculitis.

    Science.gov (United States)

    Domiciano, Talita P; Wakita, Daiko; Jones, Heather D; Crother, Timothy R; Verri, Waldiceu A; Arditi, Moshe; Shimada, Kenichi

    2017-02-02

    Interleukin-1β (IL-1β) is a highly inflammatory cytokine that significantly contributes to both acute and chronic inflammatory diseases. The secretion of IL-1β requires a unique protease, caspase-1, which is activated by various protein platforms called inflammasomes. Data suggests a key role for mitochondrial reactive oxygen species for inflammasome activation. Flavonoids constitute a group of naturally occurring polyphenolic molecules with many biological activities, including antioxidant effects. In this study, we investigated the effect of three flavonoids, quercetin (QUC), naringenin, and silymarim on inflammasome activation. We found that QUC inhibits IL-1β secretion by both the NLRP3 and AIM2 inflammasome in a dose dependent manner, but not the NLRC4 inflammasome. QUC inhibition of the inflammasome was still observed in Atg16l1 knockout macrophages, indicating that QUC's effect was autophagy independent. Since QUC inhibited both NLRP3 and AIM2 inflammasomes but not NLRC4, we assessed ASC speck formation. QUC reduced ASC speck formation and ASC oligomerization compared with controls. Additionally, QUC inhibited IL-1β in Cryopyrin-Associated Periodic Syndromes (CAPS) macrophages, where NLRP3 inflammasome is constitutively activated. In conclusion, QUC inhibits both the NLRP3 and AIM2 inflammasome by preventing ASC oligomerization and may be a potential therapeutic candidate for Kawasaki disease vasculitis and other IL-1 mediated inflammatory diseases.

  6. Structural insight into DNA binding and oligomerization of the multifunctional Cox protein of bacteriophage P2.

    Science.gov (United States)

    Berntsson, Ronnie P-A; Odegrip, Richard; Sehlén, Wilhelmina; Skaar, Karin; Svensson, Linda M; Massad, Tariq; Högbom, Martin; Haggård-Ljungquist, Elisabeth; Stenmark, Pål

    2014-02-01

    The Cox protein from bacteriophage P2 is a small multifunctional DNA-binding protein. It is involved in site-specific recombination leading to P2 prophage excision and functions as a transcriptional repressor of the P2 Pc promoter. Furthermore, it transcriptionally activates the unrelated, defective prophage P4 that depends on phage P2 late gene products for lytic growth. In this article, we have investigated the structural determinants to understand how P2 Cox performs these different functions. We have solved the structure of P2 Cox to 2.4 Å resolution. Interestingly, P2 Cox crystallized in a continuous oligomeric spiral with its DNA-binding helix and wing positioned outwards. The extended C-terminal part of P2 Cox is largely responsible for the oligomerization in the structure. The spacing between the repeating DNA-binding elements along the helical P2 Cox filament is consistent with DNA binding along the filament. Functional analyses of alanine mutants in P2 Cox argue for the importance of key residues for protein function. We here present the first structure from the Cox protein family and, together with previous biochemical observations, propose that P2 Cox achieves its various functions by specific binding of DNA while wrapping the DNA around its helical oligomer.

  7. Oligomeric structure of the MALT1 tandem Ig-like domains.

    Directory of Open Access Journals (Sweden)

    Liyan Qiu

    Full Text Available BACKGROUND: Mucosa-associated lymphoid tissue 1 (MALT1 plays an important role in the adaptive immune program. During TCR- or BCR-induced NF-κB activation, MALT1 serves to mediate the activation of the IKK (IκB kinase complex, which subsequently regulates the activation of NF-κB. Aggregation of MALT1 is important for E3 ligase activation and NF-κB signaling. PRINCIPAL FINDINGS: Unlike the isolated CARD or paracaspase domains, which behave as monomers, the tandem Ig-like domains of MALT1 exists as a mixture of dimer and tetramer in solution. High-resolution structures reveals a protein-protein interface that is stabilized by a buried surface area of 1256 Å(2 and contains numerous hydrogen and salt bonds. In conjunction with a second interface, these interactions may represent the basis of MALT1 oligomerization. CONCLUSIONS: The crystal structure of the tandem Ig-like domains reveals the oligomerization potential of MALT1 and a potential intermediate in the activation of the adaptive inflammatory pathway. ENHANCED VERSION: This article can also be viewed as an enhanced version in which the text of the article is integrated with interactive 3D representations and animated transitions. Please note that a web plugin is required to access this enhanced functionality. Instructions for the installation and use of the web plugin are available in Text S1.

  8. The oligomeric Rep protein of Mungbean yellow mosaic India virus (MYMIV) is a likely replicative helicase.

    Science.gov (United States)

    Choudhury, Nirupam Roy; Malik, Punjab Singh; Singh, Dharmendra Kumar; Islam, Mohammad Nurul; Kaliappan, Kosalai; Mukherjee, Sunil Kumar

    2006-01-01

    Geminiviruses replicate by rolling circle mode of replication (RCR) and the viral Rep protein initiates RCR by the site-specific nicking at a conserved nonamer (TAATATT downward arrow AC) sequence. The mechanism of subsequent steps of the replication process, e.g. helicase activity to drive fork-elongation, etc. has largely remained obscure. Here we show that Rep of a geminivirus, namely, Mungbean yellow mosaic India virus (MYMIV), acts as a replicative helicase. The Rep-helicase, requiring > or =6 nt space for its efficient activity, translocates in the 3'-->5' direction, and the presence of forked junction in the substrate does not influence the activity to any great extent. Rep forms a large oligomeric complex and the helicase activity is dependent on the oligomeric conformation ( approximately 24mer). The role of Rep as a replicative helicase has been demonstrated through ex vivo studies in Saccharomyces cerevisiae and in planta analyses in Nicotiana tabacum. We also establish that such helicase activity is not confined to the MYMIV system alone, but is also true with at least two other begomoviruses, viz., Mungbean yellow mosaic virus (MYMV) and Indian cassava mosaic virus (ICMV).

  9. Lipid-II Independent Antimicrobial Mechanism of Nisin Depends On Its Crowding And Degree Of Oligomerization

    Science.gov (United States)

    Prince, Ashutosh; Sandhu, Padmani; Kumar, Pankaj; Dash, Eva; Sharma, Shingarika; Arakha, Manoranjan; Jha, Suman; Akhter, Yusuf; Saleem, Mohammed

    2016-11-01

    Nisin inhibits bacterial growth by generating pores in cell membrane and interrupting cell-wall biosynthesis through specific lipid II interaction. However, the role of the hinge region and C-terminus residues of the peptide in antibacterial action of nisin is largely unknown. Here, using molecular dynamics simulations and experimental approach, we report that at high concentration regimes of nisin, interaction with phospholipids may equally deform the bacterial cell membranes even under significantly varying amounts of lipid-II. Membrane thinning, destabilization and decrease in lipid density depend on the degree of oligomerization of nisin. Growth kinetics of Bacillus subtilis and Escherichia coli interestingly show recovery by extended lag phase under low concentrations of nisin treatment while high concentrations of nisin caused decrease in cell viability as recorded by striking reduction in membrane potential and surface area. The significant changes in the dipole potential and fluorescence anisotropy were observed in negatively charged membranes in the absence of lipid-II with increasing concentration of nisin. The identical correlation of cell viability, membrane potential dissipation and morphology with the concentration regime of nisin, in both Bacillus subtilis (lipid II rich) and Escherichia coli (lipid II impoverished), hints at a non-specific physical mechanism where degree of membrane deformation depends on degree of crowding and oligomerization of nisin.

  10. SYNTHESIS AND CHARACTERIZATION OF A NOVEL REACTIVE OLIGOMERIC LADDERLIKE COPOLYMETHYL-AMINOPROPYL SILSESQUIOXANE

    Institute of Scientific and Technical Information of China (English)

    Qing-hua Duan; Tao-yi Zhang; Kui-lin Deng; Ping Xie; Rong-ben Zhang

    2005-01-01

    A novel ordered reactive oligomeric ladderlike copolymethyl-aminopropylsilsesquioxane (Me-AP-T), was successfully synthesized by an efficient hydrosilylation graft reaction of 1,1,1-trimethyl-N-2-propenyl-silanamine with a reactive oligomeric ladderlike copolymethyl-hydrosilsesquioxane (Me-H-T), which was prepared by "supramolecular chemistry-dictated stepwise coupling polymerization" process. FT-IR, 1H- and 29Si-NMR spectra show that hydrosilylation reaction proceeds completely. Moreover, 1H-NMR spectra also indicate that both the molar ratios of ≡SiMe group to Si-H and ≡ SiMe group to ≡ SiPrNH2 are approximately 1:1 respectively based on the ratio of integrated resonance areas of the different groups. XRD analysis and 29Si-NMR spectra are used for investigating the ladderlike structure of the polymers. A high Tg (128.38℃) of Me-AP-T shown in DSC measurement implies that the ordered reactive Me-AP-T macromolecules have much stiff chains.

  11. Oligomerization states of Bowman-Birk inhibitor by atomic force microscopy and computational approaches.

    Science.gov (United States)

    Silva, Luciano P; Azevedo, Ricardo B; Morais, Paulo C; Ventura, Manuel M; Freitas, Sonia M

    2005-11-15

    Several methods have been applied to study protein-protein interaction from structural and thermodynamic point of view. The present study reveals that atomic force microscopy (AFM), molecular modeling, and docking approaches represent alternative methods offering new strategy to investigate structural aspects in oligomerization process of proteinase inhibitors. The topography of the black-eyed pea trypsin/chymotrypsin inhibitor (BTCI) was recorded by AFM and compared with computational rigid-bodies docking approaches. Multimeric states of BTCI identified from AFM analysis showed globular-ellipsoidal shapes. Monomers, dimers, trimers, and hexamers were the most prominent molecular arrays observed in AFM images as evaluated by molecular volume calculations and corroborated by in silico docking and theoretical approaches. We therefore propose that BTCI adopts stable and well-packed self-assembled states in monomer-dimer-trimer-hexamer equilibrium. Although there are no correlation between specificity and packing efficiency among proteinases and proteinase inhibitors, the AFM and docked BTCI analyses suggest that these assemblies may exist in situ to play their potential function in oligomerization process.

  12. Evolutionary trace-based peptides identify a novel asymmetric interaction that mediates oligomerization in nuclear receptors.

    Science.gov (United States)

    Gu, Peili; Morgan, Daniel H; Sattar, Minawar; Xu, Xueping; Wagner, Ryan; Raviscioni, Michele; Lichtarge, Olivier; Cooney, Austin J

    2005-09-01

    Germ cell nuclear factor (GCNF) is an orphan nuclear receptor that plays important roles in development and reproduction, by repressing the expression of essential genes such as Oct4, GDF9, and BMP15, through binding to DR0 elements. Surprisingly, whereas recombinant GCNF binds to DR0 sequences as a homodimer, endogenous GCNF does not exist as a homodimer but rather as part of a large complex termed the transiently retinoid-induced factor (TRIF). Here, we use evolutionary trace (ET) analysis to design mutations and peptides that probe the molecular basis for the formation of this unusual complex. We find that GCNF homodimerization and TRIF complex formation are DNA-dependent, and ET suggests that dimerization involves key functional sites on both helix 3 and helix 11, which are located on opposing surfaces of the ligand binding domain. Targeted mutations in either helix of GCNF disrupt the formation of both the homodimer and the endogenous TRIF complex. Moreover, peptide mimetics of both of these ET-determined sites inhibit dimerization and TRIF complex formation. This suggests that a novel helix 3-helix 11 heterotypic interaction mediates GCNF interaction and would facilitate oligomerization. Indeed, it was determined that the endogenous TRIF complex is composed of a GCNF oligomer. These findings shed light on an evolutionarily selected mechanism that reveals the unusual DNA-binding, dimerization, and oligomerization properties of GCNF.

  13. Apolipoprotein E, especially apolipoprotein E4, increases the oligomerization of amyloid β peptide.

    Science.gov (United States)

    Hashimoto, Tadafumi; Serrano-Pozo, Alberto; Hori, Yukiko; Adams, Kenneth W; Takeda, Shuko; Banerji, Adrian Olaf; Mitani, Akinori; Joyner, Daniel; Thyssen, Diana H; Bacskai, Brian J; Frosch, Matthew P; Spires-Jones, Tara L; Finn, Mary Beth; Holtzman, David M; Hyman, Bradley T

    2012-10-24

    Alzheimer's disease (AD) is the most common progressive neurodegenerative disorder causing dementia. Massive deposition of amyloid β peptide (Aβ) as senile plaques in the brain is the pathological hallmark of AD, but oligomeric, soluble forms of Aβ have been implicated as the synaptotoxic component. The apolipoprotein E ε 4 (apoE ε4) allele is known to be a genetic risk factor for developing AD. However, it is still unknown how apoE impacts the process of Aβ oligomerization. Here, we found that the level of Aβ oligomers in APOE ε4/ε4 AD patient brains is 2.7 times higher than those in APOE ε3/ε3 AD patient brains, matched for total plaque burden, suggesting that apoE4 impacts the metabolism of Aβ oligomers. To test this hypothesis, we examined the effect of apoE on Aβ oligomer formation. Using both synthetic Aβ and a split-luciferase method for monitoring Aβ oligomers, we observed that apoE increased the level of Aβ oligomers in an isoform-dependent manner (E2 apoE4, increases Aβ oligomers in the brain. Higher levels of Aβ oligomers in the brains of APOE ε4/ε4 carriers compared with APOE ε3/ε3 carriers may increase the loss of dendritic spines and accelerate memory impairments, leading to earlier cognitive decline in AD.

  14. Phosphate and HEPES buffers potently affect the fibrillation and oligomerization mechanism of Alzheimer's Aβ peptide.

    Science.gov (United States)

    Garvey, Megan; Tepper, Katharina; Haupt, Caroline; Knüpfer, Uwe; Klement, Karolin; Meinhardt, Jessica; Horn, Uwe; Balbach, Jochen; Fändrich, Marcus

    2011-06-10

    The oligomerization of Aβ peptide into amyloid fibrils is a hallmark of Alzheimer's disease. Due to its biological relevance, phosphate is the most commonly used buffer system for studying the formation of Aβ and other amyloid fibrils. Investigation into the characteristics and formation of amyloid fibrils frequently relies upon material formed in vitro, predominantly in phosphate buffers. Herein, we examine the effects on the fibrillation and oligomerization mechanism of Aβ peptide that occur due solely to the influence of phosphate buffer. We reveal that significant differences in amyloid fibrillation are observed due to fibrillation being initiated in phosphate or HEPES buffer (at physiological pH and temperature). Except for the differing buffer ions, all experimental parameters were kept constant. Fibril formation was assessed using fluorescently monitored kinetic studies, microscopy, X-ray fiber diffraction and infrared and nuclear magnetic resonance spectroscopies. Based on this set up, we herein reveal profound effects on the mechanism and speed of Aβ fibrillation. The three histidine residues at positions 6, 13 and 14 of Aβ(1-40) are instrumental in these mechanistic changes. We conclude that buffer plays a more significant role in fibril formation than has been generally acknowledged.

  15. Characterization of oligomeric procyanidins and identification of quercetin glucuronide from lotus ( Nelumbo nucifera Gaertn.) seedpod.

    Science.gov (United States)

    Xiao, Jun-Song; Xie, Bi-Jun; Cao, Yan-Ping; Wu, Hua; Sun, Zhi-Da; Xiao, Di

    2012-03-21

    Procyanidins are a class of polyphenols in the plant kingdom. Lotus ( Nelumbo nucifera Gaertn.) seedpods, the inedible part of lotus and a byproduct during the production of lotus seeds, were found to be a new source rich in procyanidins. Detailed information about oligomeric procyanidins in lotus seedpods remains unknown. In this study, lotus seedpods were extracted using 60% aqueous methanol and characterized with phloroglucinolysis and liquid chromatography (mass spectrometry with an electrospray ionization source). The results indicate that the oligomeric and polymeric fraction had a mean degree of polymerization of 3.2 and 15.4, respectively, and consisted of (+)-catechin (m/z 289), gallocatechin or epigallocatechin (m/z 305), quercetin glycoside (m/z 463), quercetin glucuronide (m/z 477), procyanidin dimers (m/z 577.1), proanthocyanidin dimer gallate (m/z 593.3), prodelphinidin dimers (m/z 609.1), procyanidin trimers (m/z 865.1), etc. Quercetin glucuronide was further purified using flash chromatography and identified as quercetin-3-O-β-glucuronide by determining its exact mass using ion-trap time-of-flight mass spectrometry and ¹H and ¹³C nuclear magnetic resonance, ¹H-detected heteronuclear single-quantum coherence, and ¹H-detected heteronuclear multiple-bond correlation analyses.

  16. Redox and metal-regulated oligomeric state for human porphobilinogen synthase activation.

    Science.gov (United States)

    Sawada, N; Nagahara, N; Arisaka, F; Mitsuoka, K; Minami, M

    2011-06-01

    The oligomeric state of human porphobilinogen synthase (PBGS) [EC.4.2.1.24] is homooctamer, which consists of conformationally heterogenous subunits in the tertiary structure under air-saturated conditions. When PBGS is activated by reducing agent with zinc ion, a reservoir zinc ion coordinated by Cys(223) is transferred in the active center to be coordinated by Cys(122), Cys(124), and Cys(132) (Sawada et al. in J Biol Inorg Chem 10:199-207, 2005). The latter zinc ion serves as an electrophilic catalysis. In this study, we investigated a conformational change associated with the PBGS activation by reducing agent and zinc ion using analytical ultracentrifugation, negative staining electron microscopy, native PAGE, and enzyme activity staining. The results are in good agreement with our notion that the main component of PBGS is octamer with a few percent of hexamer and that the octamer changes spatial subunit arrangement upon reduction and further addition of zinc ion, accompanying decrease in f/f (0). It is concluded that redox-regulated PBGS activation via cleavage of disulfide bonds among Cys(122), Cys(124), and Cys(132) and coordination with zinc ion is closely linked to change in the oligomeric state.

  17. Dealuminated ZSM—5 Zeolite Catalyst for Ethylene Oligomerization to Liquid Fuels

    Institute of Scientific and Technical Information of China (English)

    NorAishahSaidinaAmin; DidiDwiAnggoro

    2002-01-01

    Ethylene oligomerization using ZSM-5 zeolite was investigated to study the role of Broensted acid sites in the formation of higher hydrocarbons,The oligomeriztion of olefins,dependent on the acidity of ZSM-5 zeolite ,is an important step in the conversion of natural gas to liquied fuels,The framework Si/Al ratio reflects the number of potential acid sites and the acid strength of the ZSM-5 catalyst,ZSM-5 with the mole ratio SiO2/Al2O3 equal to 30 was dealuminated for different periods of time according to the acidic ion-exchange method to produce ZSM-5 with various Si/Al ratios,The FT-IR analysis revealed that the integrated framework aluminum band,non-framework aluminum band,and silanol groups areas of the ZSM-5 zeolites decreased after being dealuminated,The performanc of the dealuminated zeolite was tested for ethylene oligomerization.The results demonstrated that the dealumination of ZSM-5 led to higher ethylene conversion,but the gasoline selectivity was reduced compared to the performance of a ZSM-5 zeolite ,The characterization results revealed the amount of aluminum in the zeolitic framework,the crystallinity of the ZSM-5 zeolite,and the Si/Al ration affected the formation of Broensted acid sites,The number of the Broensted acid sites on the catalyst active sites is important in the olefin conversion to liquied hydrocarbons.

  18. Oligomerization of Glycine and Alanine Catalyzed by Iron Oxides: Implications for Prebiotic Chemistry

    Science.gov (United States)

    Shanker, Uma; Bhushan, Brij; Bhattacharjee, G.; Kamaluddin

    2012-02-01

    Iron oxide minerals are probable constituents of the sediments present in geothermal regions of the primitive earth. They might have adsorbed different organic monomers (amino acids, nucleotides etc.) and catalyzed polymerization processes leading to the formation of the first living cell. In the present work we tested the catalytic activity of three forms of iron oxides (Goethite, Akaganeite and Hematite) in the intermolecular condensation of each of the amino acids glycine and L-alanine. The effect of zinc oxide and titanium dioxide on the oligomerization has also been studied. Oligomerization studies were performed for 35 days at three different temperatures 50, 90 and 120°C without applying drying/wetting cycling. The products formed were characterized by HPLC and ESI-MS techniques. All three forms of iron oxides catalyzed peptide bond formation (23.2% of gly2 and 10.65% of ala2). The reaction was monitored every 7 days. Formation of peptides was observed to start after 7 days at 50°C. Maximum yield of peptides was found after 35 days at 90°C. Reaction at 120°C favors formation of diketopiperazine derivatives. It is also important to note that after 35 days of reaction, goethite produced dimer and trimer with the highest yield among the oxides tested. We suggest that the activity of goethite could probably be due to its high surface area and surface acidity.

  19. Synthesis and Characterization of A Polynuclear Yttrium Trifluoroethoxide and Its Activity for Oligomerization of Phenyl Isocyanate

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A polynuclear yttrium trifluoroethoxide was synthesized and its activity for the oligomerization of phenyl isocyanate was tested. Reaction of anhydrous YCl3 with CF3CH2ONa in a 1:3 molar ratio in THF at room temperature, after workup, gave complex 1 as colorless crystals. The composition of complex 1 is [Y(OCH2CF3)3]6ONa2(THF)12, which was identified by elemental analysis and 1H NMR. Further X-ray structure determination reveals that complex 1 is a polynuclear ion pair compound, [Y6(μ6-O)(OCH2CF3)18]2-[Na(THF)6]+2 (1). The crystal data of complex 1 are trigonal, P-3 space group, a=1.78440(14) nm, b=1.78440(14) nm, c=1.27395(12) nm, γ=120°, V=3.5129(5) nm3, Z=1, Dc=1.607 mg·m-3, μ=2.591 mm-1 (Mo Kα), F(000)=1710, R=0.067, wR=0.182. Preliminary results demonstrated that complex 1 is a highly effective catalyst for the oligomerization of phenyl isocyanate.

  20. Influence of oligomeric silsesquioxane coating treatment on interfacial properties of CF/PAA composites

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xuezhong; HUANG Yudong; WANG Tianyu; HU Lijiang

    2007-01-01

    Carbon fibres (CF) were modified with different oligomeric silsesquioxane (SSO) coatings to improve the interfacial property of carbon fibres/polyarylacetylene (CF/PAA).The interlaminar shear strength (ILSS) of CF/PAA was tested to determine the effect of the treatment.Atomic force microscopy (AFM) in force modulation mode was adopted to study the cross-section surface of unidirectional CF/PAA composites and the relative stiffness of various phases, including CF,interphase and resin.The probability histogram and line distribution of CF/PAA cross-section surface relative stiffness,obtained from the statistical analysis of relative stiffness image,were used to compare and study the interface characterizations of composites.The results show that the ILSS increases effectively and the effects on interfacial characterizations are distinguished from each other in accor-dance with the CF surface modified with different SSO coatings owing to the various structures.Cage oligomeric silsesquioxane,including large organic groups (methacryl isobutyl-POSS),has better treatment result.AFM observa-tions lead to the conclusion that an interfacial transition layer with different morphology and stiffness appears in CF/PAA composites after being treated by the SSO coatings of different structures.It can be inferred that the appearance of the transition layer may contribute to the improvement of fibre/matrix adhesion.

  1. Development of a fast atom bombardment tandem mass spectrometric screening method for alkyl-ended oligomeric biguanides

    Science.gov (United States)

    Monaghan, J. J.; Morden, W. E.

    1992-12-01

    The FAB-MS and FAB-MS---MS behaviour of a series of "alkyl-ended" oligomeric biguanides has been studied. MS---MS product ion scans showed that fragmentation of these species occurred by predictable cleavages of the biguanide chains. Results are presented from a number of MS---MS precursor ion and neutral loss scans studied in an attempt to develop a screening method for biguanides in complex mixtures. The most effective of these scans is shown to be the neutral loss of the N-alkylguanidine molecule. Use of this scan is demonstrated for a mixture of oligomeric biguanides.

  2. Regulation of complement by cartilage oligomeric matrix protein allows for a novel molecular diagnostic principle in rheumatoid arthritis

    DEFF Research Database (Denmark)

    Happonen, Kaisa E; Saxne, Tore; Aspberg, Anders;

    2010-01-01

    Cartilage oligomeric matrix protein (COMP) is a structural component of cartilage, where it catalyzes collagen fibrillogenesis. Elevated amounts of COMP are found in serum during increased turnover of cartilage associated with active joint disease, such as rheumatoid arthritis (RA) and osteoarthr......Cartilage oligomeric matrix protein (COMP) is a structural component of cartilage, where it catalyzes collagen fibrillogenesis. Elevated amounts of COMP are found in serum during increased turnover of cartilage associated with active joint disease, such as rheumatoid arthritis (RA......) and osteoarthritis (OA). This study was undertaken to investigate the ability of COMP to regulate complement, a capacity that has previously been shown for some other cartilage proteins....

  3. Catalysis of the Oligomerization of O-Phospho-Serine, Aspartic Acid, or Glutamic Acid by Cationic Micelles

    Science.gov (United States)

    Boehler, Christof; Hill, Aubrey R., Jr.; Orgel, Leslie E.

    1996-01-01

    Treatment of relatively concentrated aqueous solutions of O-phospho-serine (50 mM), aspartic acid (100 mM) or glutamic acid (100 mM) with carbonyldiimidazole leads to the formation of an activated intermediate that oligomerizes efficiently. When the concentration of amino acid is reduced tenfold, few long oligomers can be detected. Positively-charged cetyltrimethyl ammonium bromide micelles concentrate the negatively-charged activated intermediates of the amino acids at their surfaces and catalyze efficient oligomerization even from dilute solutions.

  4. Vision-based detection of MAG weld pool

    Institute of Scientific and Technical Information of China (English)

    Gao Jinqiang; Wu Chuansong; Zhang Min; Zhao Yanhua

    2007-01-01

    Weld pool contains significant information about the welding process. The weld pool images of MAG welding are detected by LaserStrobe system. An algorithm for extracting weld pool edge is proposed according to the characteristics of MAG weld pool images. The maximum weld pool length and width are calculated. The measurement data can be used to verify the results of welding process simulation and to provide a good foundation for automatic control of MAG welding process.

  5. Pooling ASR data for closely related languages

    CSIR Research Space (South Africa)

    Van Heerden, C

    2010-05-01

    Full Text Available We describe several experiments that were conducted to assess the viability of data pooling as a means to improve speech-recognition performance for under-resourced languages. Two groups of closely related languages from the Southern Bantu language...

  6. Transferring Goods or Splitting a Resource Pool

    Science.gov (United States)

    Dijkstra, Jacob; Van Assen, Marcel A. L. M.

    2008-01-01

    We investigated the consequences for exchange outcomes of the violation of an assumption underlying most social psychological research on exchange. This assumption is that the negotiated direct exchange of commodities between two actors (pure exchange) can be validly represented as two actors splitting a fixed pool of resources (split pool…

  7. The Pool with the Movable Bottom

    Science.gov (United States)

    American School and University, 1977

    1977-01-01

    A major diagnostic, therapeutic, educational, and training center for the handicapped has under construction a swimming pool with a floor that will rise to deck level to enable handicapped persons to roll their wheel chairs on and then float free as the floor is lowered. (Author/MLF)

  8. Enhanced heat transfer in confined pool boiling

    NARCIS (Netherlands)

    Rops, C.M.; Lindken, R.; Velthuis, J.F.M.; Westerweel, J.

    2009-01-01

    We report the results of an experimental investigation of the heat transfer during nucleate boiling on a spatially confined boiling surface. The heat flux as a function of the boiling surface temperature was measured in pool boiling pots with diameters ranging from 15 mm down to 4.5 mm. It was found

  9. Enhanced heat transfer in confined pool boiling

    Energy Technology Data Exchange (ETDEWEB)

    Rops, C.M. [TNO Science and Industry, P.O. Box 155, 2600 AD Delft (Netherlands)], E-mail: cor.rops@tno.nl; Lindken, R. [Laboratory for Aero and Hydrodynamics, Delft University of Technology (Netherlands); Velthuis, J.F.M. [TNO Science and Industry, P.O. Box 155, 2600 AD Delft (Netherlands); Westerweel, J. [Laboratory for Aero and Hydrodynamics, Delft University of Technology (Netherlands)

    2009-08-15

    We report the results of an experimental investigation of the heat transfer during nucleate boiling on a spatially confined boiling surface. The heat flux as a function of the boiling surface temperature was measured in pool boiling pots with diameters ranging from 15 mm down to 4.5 mm. It was found that a reduction of the pool diameter leads to an enhancement of the nucleate boiling heat flux for most of the boiling curve. Our experimental results indicate that this enhancement is not affected by the depth of the boiling pot, the material of the bounding wall, or the diameter of the inlet water supply. High-speed camera imaging shows that the heat transfer enhancement for the spatially confined pool boiling occurs in conjunction with a stable circulating flow, which is in contrast to the chaotic and mainly upward motion for boiling in larger pool diameters. An explanation for the enhancement of the heat transfer and the associated change in flow pattern is found in the singularisation of the nucleate boiling process.

  10. "Teisele poole" Agambeniga ja Agambenita / Ragne Nukk

    Index Scriptorium Estoniae

    Nukk, Ragne, 1984-

    2012-01-01

    Tallinna Fotokuu rahvusvahelise fotonäituse "Teisele poole" ideeliste lähtekohtade tõlgendamisest. Autor kõrvutab Adam Budaki kuraatornäituse aluseks olnud Giorgio Agambeni esseed „Viimne kohtupäev“ näitusel eksponeeritud piltidega

  11. 24 CFR 320.9 - Pool administration.

    Science.gov (United States)

    2010-04-01

    ...) GOVERNMENT NATIONAL MORTGAGE ASSOCIATION, DEPARTMENT OF HOUSING AND URBAN DEVELOPMENT GUARANTY OF MORTGAGE-BACKED SECURITIES Pass-Through Type Securities § 320.9 Pool administration. The Association will only guarantee securities if the issuer executes a guaranty agreement or contractual agreement in the form...

  12. Pricing Electricity in Pools With Wind Producers

    DEFF Research Database (Denmark)

    Morales González, Juan Miguel; Conejo, A. J.; Kai Liu

    2012-01-01

    This paper considers an electricity pool that includes a significant number of wind producers and is cleared through a network-constrained auction, one day in advance and on an hourly basis. The hourly auction is formulated as a two-stage stochastic programming problem, where the first stage repr...

  13. Pooling Problems with Polynomial-Time Algorithms

    NARCIS (Netherlands)

    Haugland, Dag; Hendrix, Eligius M.T.

    2016-01-01

    The computational challenge offered by many traditional network flow models is modest, and large-scale instances can be solved fast. When the composition of the flow is part of the model, the required computation time may increase substantially. This is in particular true for the pooling problem,

  14. Enhanced heat transfer in confined pool boiling

    NARCIS (Netherlands)

    Rops, C.M.; Lindken, R.; Velthuis, J.F.M.; Westerweel, J.

    2009-01-01

    We report the results of an experimental investigation of the heat transfer during nucleate boiling on a spatially confined boiling surface. The heat flux as a function of the boiling surface temperature was measured in pool boiling pots with diameters ranging from 15 mm down to 4.5 mm. It was found

  15. Carbon Residence Times in Pedogenic Carbonate Pools

    Science.gov (United States)

    Monger, H.; Feng, Y.; Karnjanapiboonwang, A.

    2013-12-01

    Soil carbonate is a huge pool of terrestrial carbon that contains at least 930 to 940 Pg C and has influx rates on the order of 1 to 12 g CaCO3/m2/yr. Such large mass to flux ratios yield long mean residence times for carbon (e.g., 85,000 years)--assuming steady state. Like other global carbon pools, the soil carbonate pool has smaller sub-pools with higher influx rates and shorter mean residence times. For example, pedogenic carbonate in coppice dunes known to have formed since 1858 and carbonate formed on lithic artifacts in soils at archaeology sites suggests mean residence times can be as short as 120 years--again assuming steady state. Harder to assess are efflux rates as CO2 emissions or bicarbonate leaching. Some Bowen-ratio studies have nevertheless found evidence for CO2 emissions resulting from carbonate dissolution, and other studies have found evidence for bicarbonate leaching based on dissolution pipes through calcic horizons using soil morphology studies. Since an understanding of mean residence times are prerequisite for a better understanding of soil carbonate in the global carbon cycle, especially in a scenario of an expanding Aridosphere, more influx and efflux measurements are needed to evaluate the possibility of carbon sequestration by soil carbonate in hyperarid, arid, semiarid, or subhumid soils.

  16. The Pool Is Not Just for Swimming

    Science.gov (United States)

    Metzker, Andrea

    2004-01-01

    Participating in water fitness workouts is one way to benefit one's health at very little cost. If the pool at a school is used only for swimming, then the benefits of having one barely causes a ripple. When the properties of water and how humans react to water are understood and applied to water activity programs, health benefits and enjoyment…

  17. Strategies for chemically healthy public swimming pools

    DEFF Research Database (Denmark)

    Hansen, Kamilla Marie Speht

    of the strategies which can be used to achieve microbiological safe water with low levels of DBPs to ensure healthy environment for bathers. There are different approaches to achieve healthy environment in public swimming pools which in this thesis are divided into three strategies: alternatives to chlorination...

  18. The warm pool in the Indian Ocean

    Digital Repository Service at National Institute of Oceanography (India)

    Vinayachandran, P.N.; Shetye, S.R.

    ) a single maximum/minimum (northern and southern part of the Pacific warm pool and the south Indian Ocean), (iii) two maxima/minima (Arabian Sea, western equatorial Indian Ocean and Southern Bay of Bengal), and (iv) a rapid rise, a steady phase and a...

  19. "Teisele poole" Agambeniga ja Agambenita / Ragne Nukk

    Index Scriptorium Estoniae

    Nukk, Ragne, 1984-

    2012-01-01

    Tallinna Fotokuu rahvusvahelise fotonäituse "Teisele poole" ideeliste lähtekohtade tõlgendamisest. Autor kõrvutab Adam Budaki kuraatornäituse aluseks olnud Giorgio Agambeni esseed „Viimne kohtupäev“ näitusel eksponeeritud piltidega

  20. Suncatcher and cool pool. Project report

    Energy Technology Data Exchange (ETDEWEB)

    Hammond, J.

    1981-03-01

    The Suncatcher is a simple, conical solar concentrating device that captures light entering clerestory windows and directs it onto thermal storage elements at the back of a south facing living space. The cone shape and inclination are designed to capture low angle winter sunlight and to reflect away higher angle summer sunlight. It is found that winter radiation through a Suncatcher window is 40 to 50% higher than through an ordinary window, and that the average solar fraction is 59%. Water-filled steal culvert pipes used for thermal storage are found to undergo less stratification, and thus to be more effective, when located where sunlight strikes the bottom rather than the top. Five Suncatcher buildings are described. Designs are considered for 32/sup 0/, 40/sup 0/ and 48/sup 0/ north latitude, and as the latitude increases, the inclination angle of the cone should be lowered. The Cool Pool is an evaporating, shaded roof pond which thermosiphons cool water into water-filled columns within a building. Preliminary experiments indicate that the best shade design has unimpeded north sky view, good ventilation, complete summer shading, a low architectural profile, and low cost attic vent lowers work. Another series of experiments established the satisfactory performance of the Cool Pool on a test building using four water-filled cylinders, two cylinders, and two cylinders connected to the Cool Pool through a heat exchanger. Although an unshaded pool cools better at night than a shaded one, daytime heat gain far offsets this advantage. A vinyl waterbag heat exchanger was developed for use with the Cool Pool. (LEW)

  1. The Tropical Western Hemisphere Warm Pool

    Science.gov (United States)

    Wang, C.; Enfield, D. B.

    2002-12-01

    The paper describes and examines variability of the tropical Western Hemisphere warm pool (WHWP) of water warmer than 28.5oC. The WHWP is the second-largest tropical warm pool on Earth. Unlike the Eastern Hemisphere warm pool in the western Pacific, which straddles the equator, the WHWP is entirely north of the equator. At various stages of development the WHWP extends over parts of the eastern North Pacific, the Gulf of Mexico, the Caribbean, and the western tropical North Atlantic. It has a large seasonal cycle and its interannual fluctuations of area and intensity are significant. Surface heat fluxes warm the WHWP through the boreal spring to an annual maximum of SST and WHWP area in the late summer/early fall, associated with eastern North Pacific and Atlantic hurricane activities and rainfall from northern South America to the southern tier of the United States. Observations suggest that a positive ocean-atmosphere feedback operating through longwave radiation and associated cloudiness seems to operate in the WHWP. During winter preceding large warm pool, there is an alteration of the Walker and Hadley circulation cells that serves as a "tropospheric bridge" for transferring Pacific ENSO effects to the Atlantic sector and inducing initial warming of warm pool. Associated with the warm SST anomalies is a decrease in sea level pressure anomalies and an anomalous increase in atmospheric convection and cloudiness. The increase in convective activity and cloudiness results in less net longwave radiation loss from the sea surface, which then reinforces SST anomalies.

  2. Sodium reflux pool-boiler solar receiver on-sun test results

    Energy Technology Data Exchange (ETDEWEB)

    Andraka, C E; Moreno, J B; Diver, R B; Moss, T A [Oak Ridge National Lab., TN (United States)

    1992-06-01

    The efficient operation of a Stirling engine requires the application of a high heat flux to the relatively small area occupied by the heater head tubes. Previous attempts to couple solar energy to Stirling engines generally involved directly illuminating the heater head tubes with concentrated sunlight. In this study, operation of a 75-kW{sub t} sodium reflux pool-boiler solar receiver has been demonstrated and its performance characterized on Sandia's nominal 75-kW{sub t} parabolic-dish concentrator, using a cold-water gas-gap calorimeter to simulate Stirling engine operation. The pool boiler (and more generally liquid-metal reflux receivers) supplies heat to the engine in the form of latent heat released from condensation of the metal vapor on the heater head tubes. The advantages of the pool boiler include uniform tube temperature, leading to longer life and higher temperature available to the engine, and decoupling of the design of the solar absorber from the engine heater head. The two-phase system allows high input thermal flux, reducing the receiver size and losses, therefore improving system efficiency. The receiver thermal efficiency was about 90% when operated at full power and 800{degree}C. Stable sodium boiling was promoted by the addition of 35 equally spaced artificial cavities in the wetted absorber surface. High incipient boiling superheats following cloud transients were suppressed passively by the addition of small amounts of xenon gas to the receiver volume. Stable boiling without excessive incipient boiling superheats was observed under all operating conditions. The receiver developed a leak during performance evaluation, terminating the testing after accumulating about 50 hours on sun. The receiver design is reported here along with test results including transient operations, steady-state performance evaluation, operation at various temperatures, infrared thermography, x-ray studies of the boiling behavior, and a postmortem analysis.

  3. Styrene oligomerization as a molecular probe reaction for zeolite acidity: a UV-Vis spectroscopy and DFT study

    NARCIS (Netherlands)

    Buurmans, I.L.C.; Pidko, E.A.; de Groot, J.M.; Stavitski, I.; van Santen, R.A.; Weckhuysen, B.M.

    2013-01-01

    A series of H-ZSM-5 crystallites with different framework Si/Al ratios was studied by analyzing the kinetics and reaction mechanism of the oligomerization of 4-fluorostyrene as molecular probe reaction for Brønsted acidity. The formation of carbocationic species was followed by UV-Vis spectroscopy.

  4. The upstream conserved regions (UCRs) mediate homo- and hetero-oligomerization of type 4 cyclic nucleotide phosphodiesterases (PDE4s).

    Science.gov (United States)

    Xie, Moses; Blackman, Brigitte; Scheitrum, Colleen; Mika, Delphine; Blanchard, Elise; Lei, Tao; Conti, Marco; Richter, Wito

    2014-05-01

    PDE4s (type 4 cyclic nucleotide phosphodiesterases) are divided into long and short forms by the presence or absence of conserved N-terminal domains termed UCRs (upstream conserved regions). We have shown previously that PDE4D2, a short variant, is a monomer, whereas PDE4D3, a long variant, is a dimer. In the present study, we have determined the apparent molecular masses of various long and short PDE4 variants by size-exclusion chromatography and sucrose density-gradient centrifugation. Our results indicate that dimerization is a conserved property of all long PDE4 forms, whereas short forms are monomers. Dimerization is mediated by the UCR domains. Given their high sequence conservation, the UCR domains mediate not only homo-oligomerization, but also hetero-oligomerization of distinct PDE4 long forms as detected by co-immunoprecipitation assays and FRET microscopy. Endogenous PDE4 hetero-oligomers are, however, low in abundance compared with homo-dimers, revealing the presence of mechanisms that predispose PDE4s towards homo-oligomerization. Oligomerization is a prerequisite for the regulatory properties of the PDE4 long forms, such as their PKA (protein kinase A)-dependent activation, but is not necessary for PDE4 protein-protein interactions. As a result, individual PDE4 protomers may independently mediate protein-protein interactions, providing a mechanism whereby PDE4s contribute to the assembly of macromolecular signalling complexes.

  5. Disulphide bond restrains the C-terminal region of thermostable direct hemolysin during folding to promote oligomerization.

    Science.gov (United States)

    Kundu, Nidhi; Tichkule, Swapnil; Pandit, Shashi Bhushan; Chattopadhyay, Kausik

    2017-01-15

    Pore-forming toxins (PFTs) are typically produced as water-soluble monomers, which upon interacting with target cells assemble into transmembrane oligomeric pores. Vibrio parahaemolyticus thermostable direct hemolysin (TDH) is an atypical PFT that exists as a tetramer in solution, prior to membrane binding. The TDH structure highlights a core β-sandwich domain similar to those found in the eukaryotic actinoporin family of PFTs. However, the TDH structure harbors an extended C-terminal region (CTR) that is not documented in the actinoporins. This CTR remains tethered to the β-sandwich domain through an intra-molecular disulphide bond. Part of the CTR is positioned at the inter-protomer interface in the TDH tetramer. Here we show that the truncation, as well as mutation, of the CTR compromise tetrameric assembly, and the membrane-damaging activity of TDH. Our study also reveals that intra-protomer disulphide bond formation during the folding/assembly process of TDH restrains the CTR to mediate its participation in the formation of inter-protomer contact, thus facilitating TDH oligomerization. However, once tetramerization is achieved, disruption of the disulphide bond does not affect oligomeric assembly. Our study provides critical insights regarding the regulation of the oligomerization mechanism of TDH, which has not been previously documented in the PFT family.

  6. A Small-Molecule Inhibitor of Bax and Bak Oligomerization Prevents Genotoxic Cell Death and Promotes Neuroprotection.

    Science.gov (United States)

    Niu, Xin; Brahmbhatt, Hetal; Mergenthaler, Philipp; Zhang, Zhi; Sang, Jing; Daude, Michael; Ehlert, Fabian G R; Diederich, Wibke E; Wong, Eve; Zhu, Weijia; Pogmore, Justin; Nandy, Jyoti P; Satyanarayana, Maragani; Jimmidi, Ravi K; Arya, Prabhat; Leber, Brian; Lin, Jialing; Culmsee, Carsten; Yi, Jing; Andrews, David W

    2017-04-20

    Aberrant apoptosis can lead to acute or chronic degenerative diseases. Mitochondrial outer membrane permeabilization (MOMP) triggered by the oligomerization of the Bcl-2 family proteins Bax/Bak is an irreversible step leading to execution of apoptosis. Here, we describe the discovery of small-molecule inhibitors of Bax/Bak oligomerization that prevent MOMP. We demonstrate that these molecules disrupt multiple, but not all, interactions between Bax dimer interfaces thereby interfering with the formation of higher-order oligomers in the MOM, but not recruitment of Bax to the MOM. Small-molecule inhibition of Bax/Bak oligomerization allowed cells to evade apoptotic stimuli and rescued neurons from death after excitotoxicity, demonstrating that oligomerization of Bax is essential for MOMP. Our discovery of small-molecule Bax/Bak inhibitors provides novel tools for the investigation of the mechanisms leading to MOMP and will ultimately facilitate development of compounds inhibiting Bax/Bak in acute and chronic degenerative diseases. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Orally administrated cinnamon extract reduces β-amyloid oligomerization and corrects cognitive impairment in Alzheimer's disease animal models.

    Directory of Open Access Journals (Sweden)

    Anat Frydman-Marom

    Full Text Available An increasing body of evidence indicates that accumulation of soluble oligomeric assemblies of β-amyloid polypeptide (Aβ play a key role in Alzheimer's disease (AD pathology. Specifically, 56 kDa oligomeric species were shown to be correlated with impaired cognitive function in AD model mice. Several reports have documented the inhibition of Aβ plaque formation by compounds from natural sources. Yet, evidence for the ability of common edible elements to modulate Aβ oligomerization remains an unmet challenge. Here we identify a natural substance, based on cinnamon extract (CEppt, which markedly inhibits the formation of toxic Aβ oligomers and prevents the toxicity of Aβ on neuronal PC12 cells. When administered to an AD fly model, CEppt rectified their reduced longevity, fully recovered their locomotion defects and totally abolished tetrameric species of Aβ in their brain. Furthermore, oral administration of CEppt to an aggressive AD transgenic mice model led to marked decrease in 56 kDa Aβ oligomers, reduction of plaques and improvement in cognitive behavior. Our results present a novel prophylactic approach for inhibition of toxic oligomeric Aβ species formation in AD through the utilization of a compound that is currently in use in human diet.

  8. Permeability Changes of Manduca sexta Midgut Brush Border Membranes Induced by Oligomeric Structures of Different Cry Toxins

    NARCIS (Netherlands)

    Muñoz-Garay, C.; Sánchez, J.; Darszon, A.; Maagd, de R.A.; Bakker, P.; Soberón, M.; Bravo, A.

    2006-01-01

    The pore-formation activity of monomeric and oligomeric forms of different Cry1 toxins (from Cry1A to Cry1G) was analyzed by monitoring ionic permeability across Manduca sexta brush border membrane vesicles. The membrane vesicles were isolated from microvilli structures, showing a high enrichment of

  9. Recognition of coxiella burnetii by toll-like receptors and nucleotide-binding oligomerization domain-like receptors

    NARCIS (Netherlands)

    Ammerdorffer, Anne; Schoffelen, Teske; Gresnigt, Mark S.; Oosting, Marije; Brok, Den Martijn H.; Abdollahi-Roodsaz, Shahla; Kanneganti, Thirumala Devi; Jong, De Dirk J.; Deuren, Van Marcel; Roest, Hendrik Jan; Rebel, Johanna M.J.; Netea, Mihai G.; Joosten, Leo A.B.; Sprong, Tom

    2015-01-01

    Background. Infection with Coxiella burnetii can lead to acute and chronic Q fever. Toll-like receptor 1 (TLR1), TLR2, TLR4, TLR6, nucleotide-binding oligomerization domain receptor 1 (NOD1), NOD2, and the mitogen-activated protein kinases are central in the innate immune response against

  10. Recognition of Coxiella burnetii by Toll-like Receptors and Nucleotide-Binding Oligomerization Domain-like Receptors

    NARCIS (Netherlands)

    Ammerdorffer, A.; Schoffelen, T.; Gresnigt, M.S.; Oosting, M.; Brok, M.H.M.G.M. den; Abdollahi-Roodsaz, S.; Kanneganti, T.D.; Jong, D.J. de; Deuren, M. van; Roest, H.J.; Rebel, J.M.; Netea, M.G.; Joosten, L.A.B.; Sprong, T.

    2015-01-01

    BACKGROUND: Infection with Coxiella burnetii can lead to acute and chronic Q fever. Toll-like receptor 1 (TLR1), TLR2, TLR4, TLR6, nucleotide-binding oligomerization domain receptor 1 (NOD1), NOD2, and the mitogen-activated protein kinases are central in the innate immune response against

  11. Interference with RUNX1/ETO Leukemogenic Function by Cell-Penetrating Peptides Targeting the NHR2 Oligomerization Domain

    Directory of Open Access Journals (Sweden)

    Yvonne Bartel

    2013-01-01

    Full Text Available The leukemia-associated fusion protein RUNX1/ETO is generated by the chromosomal translocation t(8;21 which appears in about 12% of all de novo acute myeloid leukemias (AMLs. Essential for the oncogenic potential of RUNX1/ETO is the oligomerization of the chimeric fusion protein through the nervy homology region 2 (NHR2 within ETO. In previous studies, we have shown that the intracellular expression of peptides containing the NHR2 domain inhibits RUNX1/ETO oligomerization, thereby preventing cell proliferation and inducing differentiation of RUNX1/ETO transformed cells. Here, we show that introduction of a recombinant TAT-NHR2 fusion polypeptide into the RUNX1/ETO growth-dependent myeloid cell line Kasumi-1 results in decreased cell proliferation and increased numbers of apoptotic cells. This effect was highly specific and mediated by binding the TAT-NHR2 peptide to ETO sequences, as TAT-polypeptides containing the oligomerization domain of BCR did not affect cell proliferation or apoptosis in Kasumi-1 cells. Thus, the selective interference with NHR2-mediated oligomerization by peptides represents a challenging but promising strategy for the inhibition of the leukemogenic potential of RUNX1/ETO in t(8;21-positive leukemia.

  12. Interference with RUNX1/ETO Leukemogenic Function by Cell-Penetrating Peptides Targeting the NHR2 Oligomerization Domain

    Science.gov (United States)

    Bartel, Yvonne; Grez, Manuel; Wichmann, Christian

    2013-01-01

    The leukemia-associated fusion protein RUNX1/ETO is generated by the chromosomal translocation t(8;21) which appears in about 12% of all de novo acute myeloid leukemias (AMLs). Essential for the oncogenic potential of RUNX1/ETO is the oligomerization of the chimeric fusion protein through the nervy homology region 2 (NHR2) within ETO. In previous studies, we have shown that the intracellular expression of peptides containing the NHR2 domain inhibits RUNX1/ETO oligomerization, thereby preventing cell proliferation and inducing differentiation of RUNX1/ETO transformed cells. Here, we show that introduction of a recombinant TAT-NHR2 fusion polypeptide into the RUNX1/ETO growth-dependent myeloid cell line Kasumi-1 results in decreased cell proliferation and increased numbers of apoptotic cells. This effect was highly specific and mediated by binding the TAT-NHR2 peptide to ETO sequences, as TAT-polypeptides containing the oligomerization domain of BCR did not affect cell proliferation or apoptosis in Kasumi-1 cells. Thus, the selective interference with NHR2-mediated oligomerization by peptides represents a challenging but promising strategy for the inhibition of the leukemogenic potential of RUNX1/ETO in t(8;21)-positive leukemia. PMID:23865046

  13. The Solvent Effect on the Chemoselectivity of Palladium-catalyzed Oligomerization of 3, 3-Dimethyl-1-butyne

    Institute of Scientific and Technical Information of China (English)

    Jin Sheng CHENG; Jin Heng LI; Huan Feng JIANG; Xiao Yue OUYANG

    2003-01-01

    The chemoselectivities of PdC12 and CuC12-catalyzed oligomerization of 3, 3-dimethyl-butyne: 1, 3, 5-tri-tert-butylbenzene, 2, 2, 7, 7-tetramethyl-3, 6-dichloro-3, 5-octadiene and 2, 2, 7,7- tetramethyl-3,5-octadiyne were obtained, respectively, by regulating the polarity of the solvent.

  14. Structure of viroid replicative intermediates: physico-chemical studies on SP6 transcripts of cloned oligomeric potato spindle tuber viroid.

    Science.gov (United States)

    Steger, G; Tabler, M; Brüggemann, W; Colpan, M; Klotz, G; Sänger, H L; Riesner, D

    1986-12-22

    The structure and structural transitions of transcripts of cloned oligomeric viroid were studied in physico-chemical experiments and stability calculations. Transcripts of (+) and (-) polarity, from unit up to sixfold length, were synthesized from DNA clones of the potato spindle tuber viroid (PSTV) with the SP6 transcription system. Their structural properties were investigated by optical denaturation curves, high performance liquid chromatography (HPLC), electron microscopy, sedimentation-diffusion equilibrium and velocity sedimentation. Secondary structures of the RNAs and theoretical denaturation curves were calculated using an energy optimization program. The secondary structure of lowest free energy for unit length and oligomeric transcripts is a rod-like structure similar to that of the mature circular viroids. When this structure is used as a model for calculations, there is a large degree of agreement between the theoretical and the experimental denaturation curves. At high temperatures, however, (+) strand transcripts exhibited a transition which was more stable than expected from the calculations or than was known from curves of mature viroids. This transition arises from a rearrangement of the central conserved region of viroids to a helical region of 28 stable base pairs either intermolecularly leading to bimolecular complexes, or intramolecularly giving rise to a branched secondary structure. The rearrangement could be detected by electron microscopy, HPLC, and analytical ultracentrifugation. The helical region serves to divide up the oligomeric (+) strand into structural units which may be recognized by cleavage and ligation enzymes which process the oligomeric intermediates to circular mature viroids.

  15. A rapid, semi-quantitative assay to screen for modulators of alpha-synuclein oligomerization ex vivo

    Directory of Open Access Journals (Sweden)

    Marion eDelenclos

    2016-01-01

    Full Text Available Alpha synuclein (αsyn aggregates are associated with the pathogenesis of Parkinson’s disease and others related disorders. Although modulation of αsyn aggregation is an attractive therapeutic target, new powerful methodologies are desperately needed to facilitate in vivo screening of novel therapeutics. Here we describe an in vivo rodent model with the unique ability to rapidly track αsyn-αsyn interactions and thus oligomerization using a bioluminescent protein complementation strategy that monitors spatial and temporal αsyn oligomerization ex vivo. We find that αsyn forms oligomers in vivo as early as 1 week after stereotactic AAV injection into rat substantia nigra. Strikingly, although abundant αsyn expression is also detected in striatum at one week, no αsyn oligomers are detected at this time point. By 4 weeks, oligomerization of αsyn is detected in both striatum and substantia nigra homogenates. Moreover, in a proof-of-principle experiment, the effect of a previously described Hsp90 inhibitor known to prevent αsyn oligomer formation, demonstrates the utility of this rapid and sensitive animal model to monitor αsyn oligomerization status in the rat brain.

  16. Engineering Encounters: Reverse Engineering

    Science.gov (United States)

    McGowan, Veronica Cassone; Ventura, Marcia; Bell, Philip

    2017-01-01

    This column presents ideas and techniques to enhance your science teaching. This month's issue shares information on how students' everyday experiences can support science learning through engineering design. In this article, the authors outline a reverse-engineering model of instruction and describe one example of how it looked in our fifth-grade…

  17. Oligomerization of Vibrio cholerae Hemolysin Induces CXCR3 Upregulation and Activation of B-1a Cell

    Institute of Scientific and Technical Information of China (English)

    Gayatri Mukherjee; Kalyan K Banerjee; Tapas Biswas

    2008-01-01

    The hemolysin oligomer promotes the proliferation of B-1a cells and the expression of CD25, which is indicative of cell activation, on B-1a cells. The upregulation of CD86 induced by the oligomer showed its selective bias for the B7-2 member of B7 family while the monomer failed to induce these effects. The oligomer induced the expression of CXCR3, associated with B cell activation, while the monomer induced the expression of CXCL4, a powerful angiostatic chemokine. In conclusion, we found that B-1a cells responded to the apoptogenic monomer by expressing CXCL4, whereas oligomerization of the immunogen induced CXCR3 to shift the response towards activation. Cellular & Molecular Immunology. 2008;5(3):231-234.

  18. Efficient light-emitting devices based on platinum-complexes-anchored polyhedral oligomeric silsesquioxane materials

    KAUST Repository

    Yang, Xiaohui

    2010-08-24

    The synthesis, photophysical, and electrochemical characterization of macromolecules, consisting of an emissive platinum complex and carbazole moieties covalently attached to a polyhedral oligomeric silsesquioxane (POSS) core, is reported. Organic light-emitting devices based on these POSS materials exhibit a peak external quantum efficiency of ca. 8%, which is significantly higher than that of the analogous devices with a physical blend of the platinum complexes and a polymer matrix, and they represent noticeable improvement in the device efficiency of solution-processable phosphorescent excimer devices. Furthermore, the ratio of monomer and excimer/aggregate electroluminescent emission intensity, as well as the device efficiency, increases as the platinum complex moiety presence on the POSS macromolecules decreases. © 2010 American Chemical Society.

  19. Entropic estimate of cooperative binding of substrate on a single oligomeric enzyme: An index of cooperativity

    CERN Document Server

    Banerjee, Kinshuk; Gangopadhyay, Gautam

    2012-01-01

    Here we have systematically studied the cooperative binding of substrate molecules on the active sites of a single oligomeric enzyme in a chemiostatic condition. The average number of bound substrate and the net velocity of the enzyme catalyzed reaction are studied by the formulation of stochastic master equation for the cooperative binding classified here as spatial and temporal. We have estimated the entropy production for the cooperative binding schemes based on single trajectory analysis using a kinetic Monte Carlo technique. It is found that the total as well as the medium entropy production show the same generic diagnostic signature for detecting the cooperativity, usually characterized in terms of the net velocity of the reaction. This feature is also found to be valid for the total entropy production rate at the nonequilibrium steady state. We have introduced an index of cooperativity, C, defined in terms of the ratio of the surprisals or equivalently, the stochastic system entropy associated with the...

  20. Energetic Calculations to Decipher pH-Dependent Oligomerization and Domain Swapping of Proteins.

    Science.gov (United States)

    Shingate, Prashant; Warwicker, Jim; Sowdhamini, Ramanathan

    2015-01-01

    Domain swapping mechanism is a specialised mode of oligomerization of proteins in which part of a protein is exchanged in a non-covalent manner between constituent subunits. This mechanism is highly affected by several physiological conditions. Here, we present a detailed analysis ofthe effect of pH on different regions of the domain swapped oligomer by considering examples which are known to be sensitive to pH in transiting from monomeric to domain-swapped dimeric form. The energetic calculations were performed using a specialized method which considers changes in pH and subsequent changes in the interactions between subunits. This analysis provides definitive hints about the pH-dependence switch from monomer to domain-swapped oligomer and the steps that may be involved in the swapping mechanism.

  1. Pattern transfer during deposition and fixation of oligomeric bisphenol A on pre-structured copper surfaces.

    Science.gov (United States)

    Szillat, Florian; Fechner, Renate; Mayr, Stefan G

    2013-09-01

    Pattern transfer during deposition of oligomeric bisphenol A (OBA) on pre-structured Cu surfaces is investigated by means of a combined experimental-computational approach. Aiming for quantitative prediction of experiments, as characterized by atomic force microscopy (AFM), we explore the capabilities of stochastic rate equations to quantitatively account for the spatio-temporal evolution of surface topography. While surface diffusion and deposition noise constitute the main mechanisms, pattern transfer is affected by the inclusion of retardation in the interface potential, which appears to be necessary beyond a critical initial surface slope. In addition, routes for successful surface fixation by cross-linking are also demonstrated, which may pave the way for further technological use.

  2. The Role of Nucleotide-Binding Oligomerization Domain-Like Receptors in Pulmonary Infection.

    Science.gov (United States)

    Wiese, Kristin M; Coates, Bria M; Ridge, Karen M

    2017-08-01

    Pneumonia is caused by both viral and bacterial pathogens and is responsible for a significant health burden in the Unites States. The innate immune system is the human body's first line of defense against these pathogens. The recognition of invading pathogens via pattern recognition receptors leads to proinflammatory cytokine and chemokine production, followed by recruitment and activation of effector immune cells. The nonspecific inflammatory nature of the innate immune response can result in immunopathology that is detrimental to the host. In this review, we focus on one class of pattern recognition receptors, the nucleotide-binding oligomerization domain (NOD)-like receptors, specifically NOD1 and NOD2, and their role in host defense against viral and bacterial pathogens of the lung, including influenza, respiratory syncytial virus, Streptococcus pneumoniae, Chlamydophila pneumoniae, and Staphylococcus aureus. It is hoped that improved understanding of NOD1 and NOD2 activity in pneumonia will facilitate the development of novel therapies and promote improved patient outcomes.

  3. Building blocks of the apoptotic pore: how Bax and Bak are activated and oligomerize during apoptosis

    Science.gov (United States)

    Westphal, D; Kluck, R M; Dewson, G

    2014-01-01

    The central role of the Bcl-2 family in regulating apoptotic cell death was first identified in the 1980s. Since then, significant in-roads have been made in identifying the multiple members of this family, characterizing their form and function and understanding how their interactions determine whether a cell lives or dies. In this review we focus on the recent progress made in characterizing the proapoptotic Bcl-2 family members, Bax and Bak. This progress has resolved longstanding controversies, but has also challenged established theories in the apoptosis field. We will discuss different models of how these two proteins become activated and different ‘modes' by which they are inhibited by other Bcl-2 family members. We will also discuss novel conformation changes leading to Bak and Bax oligomerization and speculate how these oligomers might permeabilize the mitochondrial outer membrane. PMID:24162660

  4. Dissecting functions of the conserved oligomeric Golgi tethering complex using a cell-free assay.

    Science.gov (United States)

    Cottam, Nathanael P; Wilson, Katherine M; Ng, Bobby G; Körner, Christian; Freeze, Hudson H; Ungar, Daniel

    2014-01-01

    Vesicle transport sorts proteins between compartments and is thereby responsible for generating the non-uniform protein distribution along the eukaryotic secretory and endocytic pathways. The mechanistic details of specific vesicle targeting are not yet well characterized at the molecular level. We have developed a cell-free assay that reconstitutes vesicle targeting utilizing the recycling of resident enzymes within the Golgi apparatus. The assay has physiological properties, and could be used to show that the two lobes of the conserved oligomeric Golgi tethering complex play antagonistic roles in trans-Golgi vesicle targeting. Moreover, we can show that the assay is sensitive to several different congenital defects that disrupt Golgi function and therefore cause glycosylation disorders. Consequently, this assay will allow mechanistic insight into the targeting step of vesicle transport at the Golgi, and could also be useful for characterizing some novel cases of congenital glycosylation disorders.

  5. Energetic Calculations to Decipher pH-Dependent Oligomerization and Domain Swapping of Proteins.

    Directory of Open Access Journals (Sweden)

    Prashant Shingate

    Full Text Available Domain swapping mechanism is a specialised mode of oligomerization of proteins in which part of a protein is exchanged in a non-covalent manner between constituent subunits. This mechanism is highly affected by several physiological conditions. Here, we present a detailed analysis ofthe effect of pH on different regions of the domain swapped oligomer by considering examples which are known to be sensitive to pH in transiting from monomeric to domain-swapped dimeric form. The energetic calculations were performed using a specialized method which considers changes in pH and subsequent changes in the interactions between subunits. This analysis provides definitive hints about the pH-dependence switch from monomer to domain-swapped oligomer and the steps that may be involved in the swapping mechanism.

  6. Assembly of the intrinsic factor domains and oligomerization of the protein in the presence of cobalamin

    DEFF Research Database (Denmark)

    Fedosov, Sergey N; Fedosova, Natalya U; Berglund, Lars;

    2004-01-01

    oligomerized. A mixture of two fragments IF(30) + IF(20) and Cbl produced a firm complex, IF(30+20).Cbl, which could not associate to dimers. In contrast to IF(30+20).Cbl, the saturated full-length monomers IF(50).Cbl dimerized with K(d) approximately 1 microM. We suggest a two-domain organization of the full......-length protein, where two distant units, IF(30) and IF(20), can be assembled only by Cbl. They are connected by a protease-sensitive link, whose native structure is likely to be important for dimerization. However, linkage between two domains is not compulsory for Cbl binding. Advantages of the two...

  7. Inhibition of listeriolysin O oligomerization by lutein prevents Listeria monocytogenes infection.

    Science.gov (United States)

    Liu, Bowen; Teng, Zihao; Wang, Jianfeng; Lu, Gejin; Deng, Xuming; Li, Li

    2017-01-01

    The foodborne pathogenic bacterial species Listeria monocytogenes (L. monocytogenes) has caused incalculable damages to public health, and its successful infection requires various virulence factors, including Listeriolysin O (LLO). By forming pores in phagosomal membranes and even in some organelles, LLO plays an indispensable role in the ability of L. monocytogenes to escape from host immune attacks. Because of its critical role, LLO offers an appropriate therapeutic target against L. monocytogenes infection. Here, lutein, a natural small molecule existing widely in fruits and vegetables, is demonstrated as an effective inhibitor of LLO that works by blocking its oligomerization during invasion without showing significant bacteriostatic activity. Further assays applying lutein in cell culture models of invasion and in animal models showed that lutein could effectively inhibit L. monocytogenes infection. Overall, our results indicate that lutein may represent a promising and novel therapeutic agent against L. monocytogenes infection. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Synthesis and characterization of luminescent organic-inorganic hybrid nanocomposite from polyhedral oligomeric silsesquioxane

    Institute of Scientific and Technical Information of China (English)

    Yan

    2010-01-01

    A novel polyhedral oligomeric silsesquioxane (POSS)-based organic-inorganic hybrid nanocomposite (EF-POSS) was prepared by Pt-catalyzed hydrosilylation reaction of octahydridosilsesquioxane (T8H8, POSS) with a luminescent substituted acetylene (2-ethynyl-7-(4-(4-methylstyryl)styryl)-9,9-dioctyl-9H-fluorene (EF)) in high yield. The hybrid nanocomposite was soluble in common solvents such as CH2Cl2, CHCl3, THF and 1,4-dioxane. Its structure and property were characterized by FTIR, NMR, TGA, UV and PL, respectively. The results show that the hybrid nanocomposite with high thermal stability emits stable blue light as a result of photo excitation and possesses high photoluminescence quantum efficiency (φfL).

  9. Properties of PMR Polyimides Improved by Preparation of Polyhedral Oligomeric Silsesquioxane (POSS) Nanocomposites

    Science.gov (United States)

    Campbell, Sandi G.; Lee, Andre

    2005-01-01

    The field of hybrid organic-inorganic materials has grown drastically over the last several years. This interest stems from our ever-increasing ability to custom-build and control molecular structure at several length scales. This ability to control both the composition and structure of hybrid materials is sometimes broadly referred to as nanocomposite systems. One class of hybrid (organic-inorganic) nanostructured material is polyhedral oligomeric silsesquioxane (POSS), shown in the preceding diagram. The hybrid composition gives POSS materials dramatically enhanced properties relative to traditional hydrocarbons and inorganics. An important benefit of this technology is that it makes possible the formulations of nanostructured chemicals with excellent thermal and oxidative stability. This is largely due to the inorganic component.

  10. Chalcane-stilbene conjugates and oligomeric flavonoids from Chinese Dragon's Blood produced from Dracaena cochinchinensis.

    Science.gov (United States)

    Hao, Qian; Saito, Yoshinori; Matsuo, Yosuke; Li, Hai-Zhou; Tanaka, Takashi

    2015-11-01

    A detailed chemical investigation of Chinese Dragon's Blood, which is a traditional medicine produced form the red resin of Dracaena cochinchinensis, yielded two chalcane-stilbene conjugates, named cochinchinenenes G and H, together with 25 known compounds. The structures of these compounds were determined by spectroscopic examination. HPLC analysis of the resin indicated that the major constituents were a complex mixture of oligomeric polyphenols, which were detected as a broad hump on the base line of a HPLC chromatogram. (13)C NMR analysis indicated that the oligomers were mainly composed of oxygenated chalcane units. This suggestion was supported by the results of a thiol degradation experiment with mercaptoethanol, which yielded a thioether of 4-[(4-hydroxyphenyl)propyl]-3-methoxyphenol. Furthermore, methylation followed by electrospray ionization mass spectroscopic analysis of the resulting fractions established the presence of at least one heptamer of chalcane units. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Auto-oligomerization and hydration of pyrrole revealed by x-ray absorption spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Advanced Light Source; Schwartz, Craig P.; Uejio, Janel S.; Duffin, Andrew M.; England, Alice H.; Prendergast, David; Saykally, Richard J

    2009-05-29

    Near edge x-ray absorption fine structure (NEXAFS) spectra have been measured at the carbon and nitrogen K-edges of the prototypical aromatic molecule, pyrrole, both in the gas phase and when solvated in water, and compared with spectra simulated using a combination of classical molecular dynamics and first principles density functional theory in the excited state core hole approximation. The excellent agreement enabled detailed assignments. Pyrrole is highly reactive, particularly in water, and reaction products formed by the auto-oligomerization of pyrrole are identified. The solvated spectra have been measured at two different temperatures, indicating that the final states remain largely unaffected by both hydration and temperature. This is somewhat unexpected, since the nitrogen in pyrrole can donate a hydrogen bond to water.

  12. Tunable Gravimetric and Volumetric Hydrogen Storage Capacities in Polyhedral Oligomeric Silsesquioxane Frameworks.

    Science.gov (United States)

    Deshmukh, Amol; Chiu, Cheng-Chau; Chen, Yun-Wen; Kuo, Jer-Lai

    2016-09-28

    We study the hydrogen adsorption in porous frameworks composed of silsesquioxane cages linked via boron substituted aromatic structures by first-principles modeling. Such polyhedral oligomeric silsesquioxane (POSS) frameworks can be further modified by decorating them with metal atoms binding to the ring structures of the linkers. We have considered Sc- and Ti-doped frameworks which bind H2 via so-called Kubas interaction between hydrogen molecules and transition metal atoms. It will be demonstrated that the maximum H2 gravimetric capacity can be improved to more than 7.5 wt % by using longer linkers with more ring structures. However, the maximum H2 volumetric capacity can be tuned to more than 70 g/L by varying the size of silsesquioxane cages. We are optimistic that by varying the building blocks, POSS frameworks can be modified to meet the targets for the gravimetric and volumetric capacities set by the U.S. Department of Energy.

  13. Radiation-induced pink nickel oligomeric clusters in water. Pulse radiolysis study.

    Science.gov (United States)

    Hioul, Mohamed Larbi; Lin, Mingzhang; Belloni, Jacqueline; Keghouche, Nassira; Marignier, Jean-Louis

    2014-10-09

    γ-rays and pulse radiolysis of aqueous solutions of Ni(2+) ions in the presence of polyacrylate (PA(-)) and 2-propanol leads to the formation of metastable species absorbing at 540 nm that are ascribed to "pink" oligomeric clusters of a few nickel atoms only. The molar absorption coefficient is evaluated as ε540 nm = 3300 ± 300 L mol(-1) cm(-1) per Ni(0) atom. The successive steps from the reduction of Ni(2+) into Ni(+) ions to the formation of the pink clusters at 540 nm under conditions of complexation by PA(-) are investigated by pulse radiolysis. The yield of the formation of pink clusters increases markedly with the irradiation dose rate, demonstrating the occurrence of the disproportionation of the [Ni(+), PA(-)] complex after a single electron pulse. The reduction and nucleation mechanisms, including rate constants, in competition with the back oxidation by protons, particularly at low dose rate, are discussed.

  14. Understanding Controls on Wetting at Fluorinated Polyhedral Oligomeric Silsesquioxane/Polymer Surfaces.

    Science.gov (United States)

    Ye, Yi; Tian, Ming; Zhang, Chen; Du, Zhongjie; Mi, Jianguo

    2016-01-12

    Fluorinated polyhedral oligomeric silsesquioxane (F-POSS) nanoparticles have been widely used to enhance the hydrophobicity or oleophobicity of polymer films via constructing the specific micro/nanoscale roughness. In this work, we study the oleophobicity of pure and F-POSS-decorated poly(vinylidene fluoride) (PVDF) and poly(methyl methacrylate) (PMMA) films using a dynamic density functional theory approach. The role of nanoparticle size and coverage and the chemical features of F-POSS and the polymer film in the wetting behavior of diiodomethane droplets has been integrated to the remaining ratio of surface potential to quantitatively characterize the corner effect. It is shown that, on the basis of universal force field parameters, the theoretically predicted contact angles are in general agreement with the available experimental data.

  15. Oligomeric α-synuclein and β-amyloid variants as potential biomarkers for Parkinson's and Alzheimer's diseases.

    Science.gov (United States)

    Williams, Stephanie M; Schulz, Philip; Sierks, Michael R

    2016-01-01

    Oligomeric forms of α-synuclein and β-amyloid are toxic protein variants that are thought to contribute to the onset and progression of Parkinson's disease (PD) and Alzheimer's disease (AD), respectively. The detection of toxic variants in human cerebrospinal fluid (CSF) and blood has great promise for facilitating early and accurate diagnoses of these devastating diseases. Two hurdles that have impeded the use of these protein variants as biomarkers are the availability of reagents that can bind the different variants and a sensitive assay to detect their very low concentrations. We previously isolated antibody-based reagents that selectively bind two different oligomeric variants of α-synuclein and two of β-amyloid, and developed a phage-based capture enzyme-linked immunosorbent assay (ELISA) with subfemtomolar sensitivity to quantify their presence. Here, we used these reagents to show that these oligomeric α-synuclein variants are preferentially present in PD brain tissue, CSF and serum, and that the oligomeric β-amyloid variants are preferentially present in AD brain tissue, CSF, and serum. Some AD samples also had α-synuclein pathology and some PD samples also had β-amyloid pathology, and, very intriguingly, these PD cases also had a history of dementia. Detection of different oligomeric α-synuclein and β-amyloid species is an effective method for identifying tissue, CSF and sera from PD and AD samples, respectively, and samples that also contained early stages of other protein pathologies, indicating their potential value as blood-based biomarkers for neurodegenerative diseases.

  16. Uncovering the Mechanism of Forkhead-Associated Domain-Mediated TIFA Oligomerization That Plays a Central Role in Immune Responses.

    Science.gov (United States)

    Weng, Jui-Hung; Hsieh, Yin-Cheng; Huang, Chia-Chi Flora; Wei, Tong-You Wade; Lim, Liang-Hin; Chen, Yu-Hou; Ho, Meng-Ru; Wang, Iren; Huang, Kai-Fa; Chen, Chun-Jung; Tsai, Ming-Daw

    2015-10-13

    Forkhead-associated (FHA) domain is the only signaling domain that recognizes phosphothreonine (pThr) specifically. TRAF-interacting protein with an FHA domain (TIFA) was shown to be involved in immune responses by binding with TRAF2 and TRAF6. We recently reported that TIFA is a dimer in solution and that, upon stimulation by TNF-α, TIFA is phosphorylated at Thr9, which triggers TIFA oligomerization via pThr9-FHA domain binding and activates nuclear factor κB (NF-κB). However, the structural mechanism for the functionally important TIFA oligomerization remains to be established. While FHA domain-pThr binding is known to mediate protein dimerization, its role in oligomerization has not been demonstrated at the structural level. Here we report the crystal structures of TIFA (residues 1-150, with the unstructured C-terminal tail truncated) and its complex with the N-terminal pThr9 peptide (residues 1-15), which show unique features in the FHA structure (intrinsic dimer and extra β-strand) and in its interaction with the pThr peptide (with residues preceding rather than following pThr). These structural features support previous and additional functional analyses. Furthermore, the structure of the complex suggests that the pThr9-FHA domain interaction can occur only between different sets of dimers rather than between the two protomers within a dimer, providing the structural mechanism for TIFA oligomerization. Our results uncover the mechanism of FHA domain-mediated oligomerization in a key step of immune responses and expand the paradigm of FHA domain structure and function.

  17. The protective effects and underlying mechanism of an anti-oligomeric Aβ42 single-chain variable fragment antibody.

    Science.gov (United States)

    Zhang, Yuan; Chen, Xu; Liu, Jinyu; Zhang, Yingjiu

    2015-12-01

    Oligomeric Aβ42 aggregates have been identified as one of the major neurotoxic components of Alzheimer's disease (AD). Immunotherapy targeted against these Aβ42 aggregates has been proposed as an appropriate therapeutic approach for the treatment of AD. Here, we report an anti-oligomeric Aβ42 single-chain variable fragment (scFv) antibody, named MO6, obtained from the human antibody library of a healthy donor. ScFv MO6 specifically recognized and bound to the oligomeric Aβ42 (Aβ42 oligomers and immature protofibrils; 18-37 kDa), and reduced their levels mainly by blocking their formation, although scFv MO6 also induced disaggregation of Aβ42 aggregates. More importantly, scFv MO6 ameliorated or attenuated Aβ42-induced cytotoxicity and increased cell viability by up to 33%. Furthermore, scFv MO6 efficiently passed through an in vitro blood-brain barrier (BBB) model with a delivery efficiency of 66% after 60 min post-administration. ScFv MO6 is a monovalent antibody with an affinity constant (KD) of 5.2×10(-6) M for Aβ42 oligomers. Molecular docking simulations of Aβ42 to scFv MO6 revealed that the approach and specific binding of scFv MO6 to oligomeric Aβ42 aggregates was achieved by conformational recognition and directed induction, which resulted in a more dynamic adaptation of Aβ42 to scFv MO6, occurring mainly in the N-terminal (3-4), middle (12-19) and C-terminal (34-42) regions of Aβ42. This binding mode of scFv MO6 to Aβ42 explains its protective effects against oligomeric Aβ42. Our findings may be applied for the design of a smaller antibody specific for Aβ42 oligermers.

  18. Alternative heating of a municipal swimming pool

    Energy Technology Data Exchange (ETDEWEB)

    Lazzarin, R.

    1982-03-01

    Swimming pools require great amounts of energy at low temperature levels. Therefore the application of alternative heating systems is very suitable. Four different systems are taken into account: compression heat pump, absorption heat pump, motor driven heat pump with thermal recovery, and a combined system. A short thermodynamic analysis is carried out in order to evaluate operating savings. Initial cost estimates demonstrate the advantages of each proposed solution over the conventional system.

  19. Health risks of early swimming pool attendance.

    Science.gov (United States)

    Schoefer, Yvonne; Zutavern, Anne; Brockow, Inken; Schäfer, Torsten; Krämer, Ursula; Schaaf, Beate; Herbarth, Olf; von Berg, Andrea; Wichmann, H-Erich; Heinrich, Joachim

    2008-07-01

    Swimming pool attendance and exposure to chlorination by-products showed adverse health effects on children. We assessed whether early swimming pool attendance, especially baby swimming, is related to higher rates of early infections and to the development of allergic diseases. In 2003-2005, 2192 children were analysed for the 6-year follow-up of a prospective birth cohort study. Data on early swimming pool attendance, other lifestyle factors and medical history were collected by parental-administered questionnaire. Bivariate and multivariate logistic regression analyses were used to evaluate associations. Babies who did not participate in baby swimming had lower rates of infection in the 1st year of life (i) diarrhoea: OR 0.68 CI 95% 0.54-0.85; (ii) otitis media: OR 0.81 CI 95% 0.62-1.05; (iii) airway infections: OR 0.85 CI 95% 0.67-1.09. No clear association could be found between late or non-swimmers and atopic dermatitis or hay fever until the age of 6 years, while higher rates of asthma were found (OR 2.15 95% CI 1.16-3.99), however, potentially due to reverse causation. The study indicates that, in terms of infections, baby swimming might not be as harmless as commonly thought. Further evidence is needed to make conclusions if the current regulations on chlorine in Germany might not protect swimming pool attendees from an increased risk of gastrointestinal infections. In terms of developing atopic diseases there is no verifiable detrimental effect of early swimming.

  20. Labor market pooling and occupational agglomeration

    OpenAIRE

    Gabe, Todd M.; Jaison R. Abel

    2009-01-01

    This paper examines the micro-foundations of occupational agglomeration in U.S. metropolitan areas, with an emphasis on labor market pooling. Controlling for a wide range of occupational attributes, including proxies for the use of specialized machinery and for the importance of knowledge spillovers, we find that jobs characterized by a unique knowledge base exhibit higher levels of geographic concentration than do occupations with generic knowledge requirements. Further, by analyzing co-aggl...

  1. Poole-frenkel piezoconductive element and sensor

    Science.gov (United States)

    Habermehl, Scott D.

    2004-08-03

    A new class of highly sensitive piezoconductive strain sensor elements and sensors has been invented. The new elements function under conditions such that electrical conductivity is dominated by Poole-Frenkel transport. A substantial piezoconductive effect appears in this regime, allowing the new sensors to exhibit sensitivity to applied strain as much as two orders of magnitude in excess of prior art sensors based on doped silicon.

  2. Functional role of oligomerization for bacterial and plant SWEET sugar transporter family.

    Science.gov (United States)

    Xuan, Yuan Hu; Hu, Yi Bing; Chen, Li-Qing; Sosso, Davide; Ducat, Daniel C; Hou, Bi-Huei; Frommer, Wolf B

    2013-09-24

    Eukaryotic sugar transporters of the MFS and SWEET superfamilies consist of 12 and 7 α-helical transmembrane domains (TMs), respectively. Structural analyses indicate that MFS transporters evolved from a series of tandem duplications of an ancestral 3-TM unit. SWEETs are heptahelical proteins carrying a tandem repeat of 3-TM separated by a single TM. Here, we show that prokaryotes have ancestral SWEET homologs with only 3-TM and that the Bradyrhizobium japonicum SemiSWEET1, like Arabidopsis SWEET11, mediates sucrose transport. Eukaryotic SWEETs most likely evolved by internal duplication of the 3-TM, suggesting that SemiSWEETs form oligomers to create a functional pore. However, it remains elusive whether the 7-TM SWEETs are the functional unit or require oligomerization to form a pore sufficiently large to allow for sucrose passage. Split ubiquitin yeast two-hybrid and split GFP assays indicate that Arabidopsis SWEETs homo- and heterooligomerize. We examined mutant SWEET variants for negative dominance to test if oligomerization is necessary for function. Mutation of the conserved Y57 or G58 in SWEET1 led to loss of activity. Coexpression of the defective mutants with functional A. thaliana SWEET1 inhibited glucose transport, indicating that homooligomerization is necessary for function. Collectively, these data imply that the basic unit of SWEETs, similar to MFS sugar transporters, is a 3-TM unit and that a functional transporter contains at least four such domains. We hypothesize that the functional unit of the SWEET family of transporters possesses a structure resembling the 12-TM MFS structure, however, with a parallel orientation of the 3-TM unit.

  3. Aqueous SOA formation from radical oligomerization of methyl vinyl ketone (MVK) and methacrolein (MACR)

    Science.gov (United States)

    Renard, P.; Siekmann, F.; Ravier, S.; Temime-Roussel, B.; Clément, J.; Ervens, B.; Monod, A.

    2013-12-01

    It is now accepted that one of the important pathways of secondary organic aerosol (SOA) formation occurs through aqueous phase chemistry in the atmosphere. However, the chemical mechanisms leading to macromolecules are still not well understood. It was recently shown that oligomer production by OH radical oxidation in the aerosol aqueous phase from α-dicarbonyl precursors, such as methylglyoxal and glyoxal, is irreversible and fast. We have investigated the aqueous phase photooxidation of MACR and MVK, which are biogenic organic compounds derived from isoprene. Aqueous phase photooxidation of MVK and MACR was investigated in a photoreactor using photolysis of H2O2 as OH radical source. Electrospray high resolution mass spectrometry analysis of the solutions brought clear evidence for the formation of oligomer systems having a mass range of up to 1800 Da within less than 15 minutes of reaction. Highest oligomer formation rates were obtained under conditions of low dissolved oxygen, highest temperature (T = 298 K) and highest precursor initial concentrations ([MVK]0 = 20 mM). A radical mechanism of oligomerization is proposed to explain the formation of the high molecular weight products. Furthermore, we quantified the total amount of carbon present in oligomers. Kinetic parameters of the proposed oligomerization mechanism are constrained by means of a box model that is able to reproduce the temporal evolution of intermediates and products as observed in the laboratory experiments. Additional model simulations for atmospherically-relevant conditions will be presented that show the extent to which these radical processes contribute to SOA formation in the atmospheric multiphase system as compared to other aqueous phase as well as traditional SOA sources. MVK time profile (as measured by UV Spectroscopy) and mass spectra (obtained using UPLC-ESI-MS for the retention time range 0-5 min in the positive mode) at 5, 10 and 50 min of reaction (MVK 20 mM, 25° C, under

  4. Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Zhengliang L., E-mail: Leon.wu@bio-techne.com [Bio-Techne Inc., 614 McKinley Place NE, Minneapolis, MN 55413 (United States); Zhou, Hui [Gregg Hall, UNH Glycomics Center, University of New Hampshire (United States); Ethen, Cheryl M. [Bio-Techne Inc., 614 McKinley Place NE, Minneapolis, MN 55413 (United States); Reinhold, Vernon N., E-mail: Vernon.Reinhold@unh.edu [Gregg Hall, UNH Glycomics Center, University of New Hampshire (United States)

    2016-04-29

    The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. Yet to date, the structure component responsible for its virulence is still a mystery. In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. In this report, the N-glycans from both forms were released and characterized by mass spectrometry. It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. These findings were further supported by the results obtained from in vitro incorporation of azido fucose and {sup 3}H-labeled fucose using core-6 fucosyltransferase, FUT8. In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. - Graphical abstract: Proposed mechanism for how core-fucose prohibits the tetramerization of the 1918 pandemic viral neuraminidase. Only the cross section of the stalk region with two N-linked glycans are depicted for clarity. (A) Carbohydrate–carbohydrate interaction on non-fucosylated monomer allows tetramerization. (B) Core-fucosylation disrupts the interaction and prevents the tetramerization. - Highlights: • Expressed 1918 pandemic influenza viral neuraminidase has inactive and active forms. • The inactive form contains high level of core-6 fucose, while the active form lacks such modification. • Core fucose could interfere the oligomerization of the neuraminidase and thus its activation. • This discovery may explain

  5. Targeting Bax interaction sites reveals that only homo-oligomerization sites are essential for its activation

    Science.gov (United States)

    Peng, R; Tong, J-S; Li, H; Yue, B; Zou, F; Yu, J; Zhang, L

    2013-01-01

    Bax is a proapoptotic Bcl-2 family member that has a central role in the initiation of mitochondria-dependent apoptosis. However, the mechanism of Bax activation during apoptosis remains unsettled. It is believed that the activation of Bax is mediated by either dissociation from prosurvival Bcl-2 family members, or direct association with BH3-only members. Several interaction sites on Bax that mediate its interactions with other Bcl-2 family members, as well as its proapoptotic activity, have been identified in previous studies by other groups. To rigorously investigate the functional role of these interaction sites, we knocked in their respective mutants using HCT116 colon cancer cells, in which apoptosis induced by several stimuli is strictly Bax-dependent. Bax-mediated apoptosis was intact upon knock-in (KI) of K21E and D33A, which were shown to block the interaction of Bax with BH3-only activators. Apoptosis was partially reduced by KI of D68R, which impairs the interaction of Bax with prosurvival members, and S184V, a constitutively mitochondria-targeting mutant. In contrast, apoptosis was largely suppressed by KI of L70A/D71A, which blocks homo-oligomerization of Bax and its binding to prosurvival Bcl-2 family proteins. Collectively, our results suggest that the activation of endogenous Bax in HCT116 cells is dependent on its homo-oligomerization sites, but not those previously shown to interact with BH3-only activators or prosurvival proteins only. We therefore postulate that critical interaction sites yet to be identified, or mechanisms other than protein-protein interactions, need to be pursued to delineate the mechanism of Bax activation during apoptosis. PMID:23392123

  6. Defining the oligomerization state of γ-synuclein in solution and in cells.

    Science.gov (United States)

    Golebiewska, Urszula; Zurawsky, Cassandra; Scarlata, Suzanne

    2014-01-21

    γ-Synuclein is expressed at high levels in neuronal cells and in multiple invasive cancers. Like its family member α-synuclein, γ-synuclein is thought to be natively unfolded but does not readily form fibrils. The function of γ-synuclein is unknown, but we have found that it interacts strongly with the enzyme phospholipase Cβ (PLCβ), altering its interaction with G proteins. As a first step in determining its role, we have characterized its oligomerization using fluorescence homotransfer, photon-counting histogram analysis, and native gel electrophoresis. We found that when its expressed in Escherichia coli and purified, γ-synuclein appears monomeric on chromatographs under denaturing conditions, but under native conditions, it appears as oligomers of varying sizes. We followed the monomer-to-tetramer association by labeling the protein with fluorescein and following the concentration-dependent loss in fluorescence anisotropy resulting from fluorescence homotransfer. We also performed photon-counting histogram analysis at increasing concentrations of fluorescein-labeled γ-synuclein and found concentration-dependent oligomerization. Addition of PLCβ2, a strong γ-synuclein binding partner whose cellular expression is correlated with γ-synuclein, results in disruption of γ-synuclein oligomers. Similarly, its binding to lipid membranes promotes the monomer form. When we exogenously express γ-synuclein or microinject purified protein into cells, the protein appears monomeric. Our studies show that even though purified γ-synuclein form oligomers, when binding partners are present, as in cells, it dissociates to a monomer to bind these partners, which in turn may modify protein function and integrity.

  7. The R439C mutation in LMNA causes lamin oligomerization and susceptibility to oxidative stress

    Science.gov (United States)

    Verstraeten, Valerie LRM; Caputo, Sandrine; Van Steensel, Maurice AM; Duband-Goulet, Isabelle; Zinn-Justin, Sophie; Kamps, Miriam; Kuijpers, Helma JH; Östlund, Cecilia; Worman, Howard J; Briedé, Jacob J; Le Dour, Caroline; Marcelis, Carlo LM; Van Geel, Michel; Steijlen, Peter M; Van Den Wijngaard, Arthur; Ramaekers, Frans CS; Broers, Jos LV

    2009-01-01

    Abstract Dunnigan-type familial partial lipodystrophy (FPLD) is a laminopathy characterized by an aberrant fat distribution and a metabolic syndrome for which oxidative stress has recently been suggested as one of the disease-causing mechanisms. In a family affected with FPLD, we identified a heterozygous missense mutation c.1315C>T in the LMNA gene leading to the p.R439C substitution. Cultured patient fibroblasts do not show any prelamin A accumulation and reveal honeycomb-like lamin A/C formations in a significant percentage of nuclei. The mutation affects a region in the C-terminal globular domain of lamins A and C, different from the FPLD-related hot spot. Here, the introduction of an extra cysteine allows for the formation of disulphide-mediated lamin A/C oligomers. This oligomerization affects the interaction properties of the C-terminal domain with DNA as shown by gel retardation assays and causes a DNA-interaction pattern that is distinct from the classical R482W FPLD mutant. Particularly, whereas the R482W mutation decreases the binding efficiency of the C-terminal domain to DNA, the R439C mutation increases it. Electron spin resonance spectroscopy studies show significantly higher levels of reactive oxygen species (ROS) upon induction of oxidative stress in R439C patient fibroblasts compared to healthy controls. This increased sensitivity to oxidative stress seems independent of the oligomerization and enhanced DNA binding typical for R439C, as both the R439C and R482W mutants show a similar and significant increase in ROS upon induction of oxidative stress by H2O2. PMID:19220582

  8. Genetic analysis and serum level of cartilage oligomeric matrix protein in patients with pseudoachondroplasia

    Institute of Scientific and Technical Information of China (English)

    LIU Feng-xia; LI Zhi-ling; WEI Zhen-ji; MENG yan; REN Cui-ai; ZHANG Xu-de; YU Meng-xue; HUANG Shang-zhi

    2010-01-01

    Background Pseudoachondroplasia (PSACH) is an autosomal-dominant osteochondrodysplasia due to mutations in the gene encoding cartilage oligomeric matrix protein (COMP).Clinical diagnosis of PSACH is based primarily on family history, physical examination, and radiographic evaluation.There is evidence that decreased serum COMP concentration may serve as a diagnostic marker in PSACH.Here, we investigated the role of this gene and the serum COMP concentration in Chinese patients with PSACH.Methods A family with three patients and a sporadic case were recruited.Genomic and phenotypic data were recorded.The diagnosis of PSACH was made on the base of clinical evaluation.The genomic DNA was extracted from peripheral blood leukocytes.The 8-19 exons and flanking intron-exon boundary sequences of COMP were amplified by polymerase chain reaction (PCR) and screened for mutation by direct DNA sequencing.Serum COMP concentrations of 4 patients and age-compatible control group of 20 unrelated healthy subjects were analyzed on the basis of an ELISA Kit for human cartilage oligomeric matrix protein.Results A deletion (c.1447-1455del) was identified in exon 13 in the sporadic case.The mean serum COMP concentrations of four patients (3.12±2.28) were significantly lower than those of control group (10.86±2.21, P <0.05).There was no overlap in the distribution of serum COMP concentration between PSACH patients and controls.Conclusions Mutations in COMP gene are responsible for the PSACH.Serum COMP concentration may be suggested as an additional diagnostic marker to aid clinical findings in suspected cases of PSACH.

  9. Following activation of the amyloid cascade, apolipoprotein E4 drives the in vivo oligomerization of amyloid-β resulting in neurodegeneration.

    Science.gov (United States)

    Belinson, Haim; Kariv-Inbal, Zehavit; Kayed, Rakez; Masliah, Eliezer; Michaelson, Daniel M

    2010-01-01

    According to the amyloid hypothesis, the accumulation of oligomerized amyloid-β (Aβ) is a primary event in the pathogenesis of Alzheimer's disease (AD). The trigger of the amyloid cascade and of Aβ oligomerization in sporadic AD, the most prevalent form of the disease, remains elusive. Here, we examined the hypothesis that apolipoprotein E4 (ApoE4), the most prevalent genetic risk factor for AD, triggers the accumulation of intraneuronal oligomerized Aβ following activation of the amyloid cascade. We investigated the intracellular organelles that are targeted by these processes and govern their pathological consequences. This revealed that activation of the amyloid cascade in vivo by inhibition of the Aβ degrading enzyme neprilysin specifically results in accumulation of Aβ and oligomerized Aβ and of ApoE4 in the CA1 neurons of ApoE4 mice. This was accompanied by lysosomal and mitochondrial pathology and the co-localization of Aβ, oligomerized Aβ, and ApoE4 with enlarged lysosomes and of Aβ and oligomerized Aβ with mitochondria. The time course of the lysosomal effects paralleled that of the loss of CA1 neurons, whereas the mitochondrial effects reached an earlier plateau. These findings suggest that ApoE4 potentiates the pathological effects of Aβ and the amyloid cascade by triggering the oligomerization of Aβ, which in turn, impairs intraneuronal mitochondria and lysosomes and drives neurodegeneration.

  10. Ulk4 Regulates Neural Stem Cell Pool.

    Science.gov (United States)

    Liu, Min; Guan, Zhenlong; Shen, Qin; Flinter, Frances; Domínguez, Laura; Ahn, Joo Wook; Collier, David A; O'Brien, Timothy; Shen, Sanbing

    2016-09-01

    The size of neural stem cell (NSC) pool at birth determines the starting point of adult neurogenesis. Aberrant neurogenesis is associated with major mental illness, in which ULK4 is proposed as a rare risk factor. Little is known about factors regulating the NSC pool, or function of the ULK4. Here, we showed that Ulk4(tm1a/tm1a) mice displayed a dramatically reduced NSC pool at birth. Ulk4 was expressed in a cell cycle-dependent manner and peaked in G2/M phases. Targeted disruption of the Ulk4 perturbed mid-neurogenesis and significantly reduced cerebral cortex in postnatal mice. Pathway analyses of dysregulated genes in Ulk4(tm1a/tm1a) mice revealed Ulk4 as a key regulator of cell cycle and NSC proliferation, partially through regulation of the Wnt signaling. In addition, we identified hemizygous deletion of ULK4 gene in 1.2/1,000 patients with pleiotropic symptoms including severe language delay and learning difficulties. ULK4, therefore, may significantly contribute to neurodevelopmental, neuropsychiatric, and neurodegenerative disorders. Stem Cells 2016;34:2318-2331.

  11. Forced pooling and mediation: the odd couple

    Energy Technology Data Exchange (ETDEWEB)

    Pascoe, D.

    2001-07-01

    The mandate and activities of the office of the Ontario Mining and Lands Commissioner is explained. The office has jurisdiction under the Mining Act, the Conservation Authorities Act, the Assessment Act, the Aggregates Resources Act , the Lakes and Rivers Improvement Act, and most importantly, under the Oil, Gas and Salt Resources Act. Under the Oil, Gas and Salt Resources Act, the Commissioner may under clause (a) order the joining of interest within a spacing unit for the purpose of drilling and operating a well and apportioning the associated costs and benefits. Under clause (b) the Commissioner may, by order, require and regulate the joining of interests within a field or a pool for the purpose of drilling or operating wells and may order also the designation of management and apportioning of costs. Pooling (matters relating to spacing units) and unitization (matters relating to joining interests in a field or pool) are legislated departures from the old rule of capture which essentially allowed any landowner to sink a well on his land and remove the resource without consideration of the interest of surrounding landowners. In essence, the Mining and Land Commissioner protects the interest of all landowners within a spacing unit, while ensuring that the resource extraction can proceed under the economically and technically most favorable conditions.

  12. Vesicle Pools: Lessons from Adrenal Chromaffin Cells

    Directory of Open Access Journals (Sweden)

    David R Stevens

    2011-02-01

    Full Text Available The adrenal chromaffin cell serves as a model system to study fast Ca2+-dependent exocytosis. Membrane capacitance measurements in combination with Ca2+ uncaging offers a temporal resolution in the millisecond range and reveals that catecholamine release occurs in three distinct phases. Release of a readily releasable (RRP and a slowly releasable (SRP pool are followed by sustained release, due to maturation and release of vesicles which were not release-ready at the start of the stimulus. Trains of depolarizations, a more physiological stimulus, induce release from a small immediately releasable pool of vesicles residing adjacent to calcium channels, as well as from the RRP. The SRP is poorly activated by depolarization. A sequential model, in which non-releasable docked vesicles are primed to a slowly releasable state, and then further mature to the readily releasable state, has been proposed. The docked state, dependent on membrane proximity, requires SNAP-25, synaptotagmin and syntaxin. The ablation or modification of SNAP-25 and syntaxin, components of the SNARE complex, as well as of synaptotagmin, the calcium sensor, and modulators such complexins and Snapin alter the properties and/or magnitudes of different phases of release, and in particular can ablate the RRP. These results indicate that the composition of the SNARE complex and its interaction with modulatory molecules drives priming and provides a molecular basis for different pools of releasable vesicles.

  13. A High Diversity in Chitinolytic and Chitosanolytic Species and Enzymes and Their Oligomeric Products Exist in Soil with a History of Chitin and Chitosan Exposure.

    Science.gov (United States)

    Nampally, Malathi; Rajulu, M B Govinda; Gillet, Dominique; Suryanarayanan, T S; Moerschbacher, Bruno B

    2015-01-01

    Chitin is one of the most abundant biomolecules on earth, and its partially de-N-acetylated counterpart, chitosan, is one of the most promising biotechnological resources due to its diversity in structure and function. Recently, chitin and chitosan modifying enzymes (CCMEs) have gained increasing interest as tools to engineer chitosans with specific functions and reliable performance in biotechnological and biomedical applications. In a search for novel CCME, we isolated chitinolytic and chitosanolytic microorganisms from soils with more than ten-years history of chitin and chitosan exposure and screened them for chitinase and chitosanase isoenzymes as well as for their patterns of oligomeric products by incubating their secretomes with chitosan polymers. Of the 60 bacterial strains isolated, only eight were chitinolytic and/or chitosanolytic, while 20 out of 25 fungal isolates were chitinolytic and/or chitosanolytic. The bacterial isolates produced rather similar patterns of chitinolytic and chitosanolytic enzymes, while the fungal isolates produced a much broader range of different isoenzymes. Furthermore, diverse mixtures of oligosaccharides were formed when chitosan polymers were incubated with the secretomes of select fungal species. Our study indicates that soils with a history of chitin and chitosan exposure are a good source of novel CCME for chitosan bioengineering.

  14. Inhibiting α-synuclein oligomerization by stable cell-penetrating β-synuclein fragments recovers phenotype of Parkinson's disease model flies.

    Directory of Open Access Journals (Sweden)

    Ronit Shaltiel-Karyo

    Full Text Available The intracellular oligomerization of α-synuclein is associated with Parkinson's disease and appears to be an important target for disease-modifying treatment. Yet, to date, there is no specific inhibitor for this aggregation process. Using unbiased systematic peptide array analysis, we identified molecular interaction domains within the β-synuclein polypeptide that specifically binds α-synuclein. Adding such peptide fragments to α-synuclein significantly reduced both amyloid fibrils and soluble oligomer formation in vitro. A retro-inverso analogue of the best peptide inhibitor was designed to develop the identified molecular recognition module into a drug candidate. While this peptide shows indistinguishable activity as compared to the native peptide, it is stable in mouse serum and penetrates α-synuclein over-expressing cells. The interaction interface between the D-amino acid peptide and α-synuclein was mapped by Nuclear Magnetic Resonance spectroscopy. Finally, administering the retro-inverso peptide to a Drosophila model expressing mutant A53T α-synuclein in the nervous system, resulted in a significant recovery of the behavioral abnormalities of the treated flies and in a significant reduction in α-synuclein accumulation in the brains of the flies. The engineered retro-inverso peptide can serve as a lead for developing a novel class of therapeutic agents to treat Parkinson's disease.

  15. POOL WATER TREATMENT AND COOLING SYSTEM DESCRIPTION DOCUMENT

    Energy Technology Data Exchange (ETDEWEB)

    V. King

    2000-06-19

    The Pool Water Treatment and Cooling System is located in the Waste Handling Building (WHB), and is comprised of various process subsystems designed to support waste handling operations. This system maintains the pool water temperature within an acceptable range, maintains water quality standards that support remote underwater operations and prevent corrosion, detects leakage from the pool liner, provides the capability to remove debris from the pool, controls the pool water level, and helps limit radiological exposure to personnel. The pool structure and liner, pool lighting, and the fuel staging racks in the pool are not within the scope of the Pool Water Treatment and Cooling System. Pool water temperature control is accomplished by circulating the pool water through heat exchangers. Adequate circulation and mixing of the pool water is provided to prevent localized thermal hotspots in the pool. Treatment of the pool water is accomplished by a water treatment system that circulates the pool water through filters, and ion exchange units. These water treatment units remove radioactive and non-radioactive particulate and dissolved solids from the water, thereby providing the water clarity needed to conduct waste handling operations. The system also controls pool water chemistry to prevent advanced corrosion of the pool liner, pool components, and fuel assemblies. Removal of radioactivity from the pool water contributes to the project ALARA (as low as is reasonably achievable) goals. A leak detection system is provided to detect and alarm leaks through the pool liner. The pool level control system monitors the water level to ensure that the minimum water level required for adequate radiological shielding is maintained. Through interface with a demineralized water system, adequate makeup is provided to compensate for loss of water inventory through evaporation and waste handling operations. Interface with the Site Radiological Monitoring System provides continuous

  16. UMRS LTRMP 2010/11 LCU Mapping -- Pool 21

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of...

  17. UMRS LTRMP 2010/11 LCU Mapping -- Pool 3

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of...

  18. UMRS LTRMP 2010/11 LCU Mapping -- Pool 6

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of...

  19. UMRS LTRMP 2010/11 LCU Mapping -- Pool 14

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  20. UMRS LTRMP 2010/11 LCU Mapping -- Pool 7

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  1. UMRS LTRMP 2010/11 LCU Mapping -- Pool 6

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  2. UMRS LTRMP 2010/11 LCU Mapping -- Pool 10

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  3. UMRS LTRMP 2010/11 LCU Mapping -- Pool 4

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  4. UMRS LTRMP 2010/11 LCU Mapping -- Pool 13

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  5. UMRS LTRMP 2010/11 LCU Mapping -- Pool 22

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  6. UMRS LTRMP 2010/11 LCU Mapping -- Pool 18

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  7. UMRS LTRMP 2010/11 LCU Mapping -- Pool 5a

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  8. UMRS LTRMP 2010/11 LCU Mapping -- Pool 3

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  9. UMRS LTRMP 2010/11 LCU Mapping -- Pool 8

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  10. UMRS LTRMP 2010/11 LCU Mapping -- Pool 24

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  11. UMRS LTRMP 2010/11 LCU Mapping -- Pool 19

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  12. UMRS LTRMP 2010/11 LCU Mapping -- Pool 20

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  13. UMRS LTRMP 2010/11 LCU Mapping -- Pool 26

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  14. UMRS LTRMP 2010/11 LCU Mapping -- Pool 09

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  15. UMRS LTRMP 2010/11 LCU Mapping -- Pool 25

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  16. UMRS LTRMP 2010/11 LCU Mapping -- Pool 21

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  17. UMRS LTRMP 2010/11 LCU Mapping -- Pool 12

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  18. UMRS LTRMP 2010/11 LCU Mapping -- Pool 5

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of 2010...

  19. UMRS LTRMP 2010/11 LCU Mapping -- Pool 25

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Aerial photographs for Pools 1-13 Upper Mississippi River System and Pools, Alton-Marseilles, Illinois River were collected in color infrared (CIR) in August of...

  20. Performance Study and Dynamic Optimization Design for Thread Pool Systems

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Dongping [Iowa State Univ., Ames, IA (United States)

    2004-12-19

    Thread pools have been widely used by many multithreaded applications. However, the determination of the pool size according to the application behavior still remains problematic. To automate this process, in this thesis we have developed a set of performance metrics for quantitatively analyzing thread pool performance. For our experiments, we built a thread pool system which provides a general framework for thread pool research. Based on this simulation environment, we studied the performance impact brought by the thread pool on different multithreaded applications. Additionally, the correlations between internal characterizations of thread pools and their throughput were also examined. We then proposed and evaluated a heuristic algorithm to dynamically determine the optimal thread pool size. The simulation results show that this approach is effective in improving overall application performance.

  1. [Tiit Hennoste loengusarjast 'Hüpped modernismi poole'] / Hasso Krull

    Index Scriptorium Estoniae

    Krull, Hasso, 1964-

    1997-01-01

    Tiit Hennoste loengusarjast 'Hüpped modernismi poole : Eesti 20. sajandi kirjandusest Euroopa modernismi taustal' (1993, nr. 10 - 1997, nr. 10/11).Vastukaja: Hüpped modernismi poole: kajad ja vastukajad // Vikerkaar (1998) nr. 6, lk. 99-111

  2. [Tiit Hennoste loengusarjast 'Hüpped modernismi poole'] / Hasso Krull

    Index Scriptorium Estoniae

    Krull, Hasso, 1964-

    1997-01-01

    Tiit Hennoste loengusarjast 'Hüpped modernismi poole : Eesti 20. sajandi kirjandusest Euroopa modernismi taustal' (1993, nr. 10 - 1997, nr. 10/11).Vastukaja: Hüpped modernismi poole: kajad ja vastukajad // Vikerkaar (1998) nr. 6, lk. 99-111

  3. Understanding the Impacts of Dark Pools on Price Discovery

    OpenAIRE

    Ye, Linlin

    2016-01-01

    This paper investigates the impact of dark pools on price discovery (the efficiency of prices on stock exchanges to aggregate information). Assets are traded in either an exchange or a dark pool, with the dark pool offering better prices but lower execution rates. Informed traders receive noisy and heterogeneous signals about an asset's fundamental. We find that informed traders use dark pools to mitigate their information risk and there is a sorting effect: in equilibrium, traders with stron...

  4. Layered Systems Engineering Engines

    Science.gov (United States)

    Breidenthal, Julian C.; Overman, Marvin J.

    2009-01-01

    A notation is described for depicting the relationships between multiple, contemporaneous systems engineering efforts undertaken within a multi-layer system-of-systems hierarchy. We combined the concepts of remoteness of activity from the end customer, depiction of activity on a timeline, and data flow to create a new kind of diagram which we call a "Layered Vee Diagram." This notation is an advance over previous notations because it is able to be simultaneously precise about activity, level of granularity, product exchanges, and timing; these advances provide systems engineering managers a significantly improved ability to express and understand the relationships between many systems engineering efforts. Using the new notation, we obtain a key insight into the relationship between project duration and the strategy selected for chaining the systems engineering effort between layers, as well as insights into the costs, opportunities, and risks associated with alternate chaining strategies.

  5. Metabolically engineered cells for the production of resveratrol or an oligomeric or glycosidically-bound derivative thereof

    DEFF Research Database (Denmark)

    2006-01-01

    A recombinant micro-organism producing resveratrol by a pathway in which phenylalanine ammonia lyase (PAL) produces trans-cinnamic acid from phenylalanine, cinnamate 4- hydroxylase (C4H) produces 4-coumaric acid from said trans- cinnamic acid, 4-coumarate-CoA ligase (4CL) produces 4- coumaroyl Co......A from said 4-coumaric acid, and resveratrol synthase (VST) produces said resveratrol from said 4- coumaroyl CoA, or in which L-phenylalanine- or tyrosine- ammonia lyase (PAL/TAL) produces 4-coumaric acid, 4- coumarate-CoA ligase (4CL) produces 4-coumaroyl CoA from said 4-coumaric acid, and resveratrol...... synthase (VST) produces said resveratrol from said 4-coumaroyl CoA. The micro-organism may be a yeast, fungus or bacterium including Saccharomyces cerevisiae, E. coli, Lactococcus lactis, Aspergillus niger, or Aspergillus oryzae....

  6. A three-pool model dissecting readily releasable pool replenishment at the calyx of held.

    Science.gov (United States)

    Guo, Jun; Ge, Jian-long; Hao, Mei; Sun, Zhi-cheng; Wu, Xin-sheng; Zhu, Jian-bing; Wang, Wei; Yao, Pan-tong; Lin, Wei; Xue, Lei

    2015-03-31

    Although vesicle replenishment is critical in maintaining exo-endocytosis recycling, the underlying mechanisms are not well understood. Previous studies have shown that both rapid and slow endocytosis recycle into a very large recycling pool instead of within the readily releasable pool (RRP), and the time course of RRP replenishment is slowed down by more intense stimulation. This finding contradicts the calcium/calmodulin-dependence of RRP replenishment. Here we address this issue and report a three-pool model for RRP replenishment at a central synapse. Both rapid and slow endocytosis provide vesicles to a large reserve pool (RP) ~42.3 times the RRP size. When moving from the RP to the RRP, vesicles entered an intermediate pool (IP) ~2.7 times the RRP size with slow RP-IP kinetics and fast IP-RRP kinetics, which was responsible for the well-established slow and rapid components of RRP replenishment. Depletion of the IP caused the slower RRP replenishment observed after intense stimulation. These results establish, for the first time, a realistic cycling model with all parameters measured, revealing the contribution of each cycling step in synaptic transmission. The results call for modification of the current view of the vesicle recycling steps and their roles.

  7. 40 CFR 97.143 - Compliance supplement pool.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Compliance supplement pool. 97.143... Allocations § 97.143 Compliance supplement pool. (a) In addition to the CAIR NOX allowances allocated under... CAIR NOX allowances to CAIR NOX units in the respective State: State Compliancesupplement pool...

  8. 40 CFR 96.143 - Compliance supplement pool.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Compliance supplement pool. 96.143... CAIR NOX Allowance Allocations § 96.143 Compliance supplement pool. (a) In addition to the CAIR NOX... Compliancesupplement pool Alabama 10,166 Delaware 843 District Of Columbia 0 Florida 8,335 Georgia 12,397 Illinois...

  9. 40 CFR 97.43 - Compliance Supplement Pool.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Compliance Supplement Pool. 97.43... Allocations § 97.43 Compliance Supplement Pool. (a) For any NOX Budget unit that reduces its NOX emission rate... State's compliance supplement pool set forth in appendix D of this part has a number of NOX...

  10. 48 CFR 873.114 - Best value pool.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false Best value pool. 873.114... SUPPLEMENTARY REGULATIONS SIMPLIFIED ACQUISITION PROCEDURES FOR HEALTH-CARE RESOURCES 873.114 Best value pool... solicitation. These vendors constitute the best value pool. This determination is within the sole discretion...

  11. 48 CFR 252.232-7000 - Advance payment pool.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 3 2010-10-01 2010-10-01 false Advance payment pool. 252... Provisions And Clauses 252.232-7000 Advance payment pool. As prescribed in 232.412-70(a), use the following clause: Advance Payment Pool (DEC 1991) (a) Notwithstanding any other provision of this contract,...

  12. New algebraic constructions for pooling design in DNA library screening.

    Science.gov (United States)

    Li, Zengti; Gao, Suogang; Du, Hongjie; Shi, Yan

    2010-01-01

    Pooling design is an important mathematical tool in DNA library screening. It has been showed that using pooling design, the number of tests in DNA library screening can be greatly reduced. In this paper, we present some new algebraic constructions for pooling design.

  13. Spatial and temporal variation in ephemeral pool crustacean communities

    Science.gov (United States)

    Janette Holtz; Marie A. Simovich; Thomas Philippi

    2005-01-01

    Vernal pool habitat losses in San Diego County, California, are estimated at 95 percent and will increase as development continues. The majority of San Diego’s remaining pools are located at the Marine Corps Air Station, Miramar. Crustacean communities in eight vernal pools in San Diego County were sampled over 3 years that differed in rainfall amount and pattern. In...

  14. 7 CFR 985.57 - Reserve pool requirements.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Reserve pool requirements. 985.57 Section 985.57... Reserve pool requirements. (a) On November 1, or such other date as the Committee, with the approval of the Secretary may establish, the Committee shall pool identified excess oil as reserve oil in...

  15. Soil organic carbon pools in the northern circumpolar permafrost region

    Science.gov (United States)

    C. Tarnocai; J.G. Canadell; E.A.G. Schuur; P. Kuhry; G. Mazhitova; S. Zimov

    2009-01-01

    The Northern Circumpolar Soil Carbon Database was developed to determine carbon pools in soils of the northern circumpolar permafrost region. Here we report a new estimate of the carbon pools in soils of the northern permafrost region, including deeper layers and pools not accounted for in previous analyses.

  16. A Skeleton for Distributed Work Pools in Eden

    DEFF Research Database (Denmark)

    Dieterle, Mischa; Berthold, Jost; Loogen, Rita

    2010-01-01

    We present a flexible skeleton for implementing distributed work pools in our parallel functional language Eden. The skeleton manages a pool of tasks (work pool) in a distributed manner using a demand-driven work stealing approach for load balancing. All coordination is done locally within...

  17. NACUBO's Guide to Unitizing Investment Pools. Second Edition

    Science.gov (United States)

    Wheeler, Mary S.

    2011-01-01

    The National Association of College and University Business Officers' (NACUBO's) "Guide to Unitizing Investment Pools" addresses the principles and concepts for administering a consolidated investment pool. Unitization is the mechanism by which investment funds are pooled to maximize investment efficiencies and provide information for donors,…

  18. Engineer Ethics

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Dae Sik; Kim, Yeong Pil; Kim, Yeong Jin

    2003-03-15

    This book tells of engineer ethics such as basic understanding of engineer ethics with history of engineering as a occupation, definition of engineering and specialized job and engineering, engineer ethics as professional ethics, general principles of ethics and its limitation, ethical theory and application, technique to solve the ethical problems, responsibility, safety and danger, information engineer ethics, biotechnological ethics like artificial insemination, life reproduction, gene therapy and environmental ethics.

  19. In situ genetic correction of the sickle cell anemia mutation in human induced pluripotent stem cells using engineered zinc finger nucleases.

    Science.gov (United States)

    Sebastiano, Vittorio; Maeder, Morgan L; Angstman, James F; Haddad, Bahareh; Khayter, Cyd; Yeo, Dana T; Goodwin, Mathew J; Hawkins, John S; Ramirez, Cherie L; Batista, Luis F Z; Artandi, Steven E; Wernig, Marius; Joung, J Keith

    2011-11-01

    The combination of induced pluripotent stem cell (iPSC) technology and targeted gene modification by homologous recombination (HR) represents a promising new approach to generate genetically corrected, patient-derived cells that could be used for autologous transplantation therapies. This strategy has several potential advantages over conventional gene therapy including eliminating the need for immunosuppression, avoiding the risk of insertional mutagenesis by therapeutic vectors, and maintaining expression of the corrected gene by endogenous control elements rather than a constitutive promoter. However, gene targeting in human pluripotent cells has remained challenging and inefficient. Recently, engineered zinc finger nucleases (ZFNs) have been shown to substantially increase HR frequencies in human iPSCs, raising the prospect of using this technology to correct disease causing mutations. Here, we describe the generation of iPSC lines from sickle cell anemia patients and in situ correction of the disease causing mutation using three ZFN pairs made by the publicly available oligomerized pool engineering method (OPEN). Gene-corrected cells retained full pluripotency and a normal karyotype following removal of reprogramming factor and drug-resistance genes. By testing various conditions, we also demonstrated that HR events in human iPSCs can occur as far as 82 bps from a ZFN-induced break. Our approach delineates a roadmap for using ZFNs made by an open-source method to achieve efficient, transgene-free correction of monogenic disease mutations in patient-derived iPSCs. Our results provide an important proof of principle that ZFNs can be used to produce gene-corrected human iPSCs that could be used for therapeutic applications.

  20. Swimming Pools, Swimming pools, licensed - name, address, contact, volume, Published in 2006, Iowa Dept. of Public Health.

    Data.gov (United States)

    NSGIC GIS Inventory (aka Ramona) — This Swimming Pools dataset, was produced all or in part from Published Reports/Deeds information as of 2006. It is described as 'Swimming pools, licensed - name,...

  1. Dominant factors in controlling marine gas pools in South China

    Science.gov (United States)

    Xu, S.; Watney, W.L.

    2007-01-01

    In marine strata from Sinian to Middle Triassic in South China, there develop four sets of regional and six sets of local source rocks, and ten sets of reservoir rocks. The occurrence of four main formation periods in association with five main reconstruction periods, results in a secondary origin for the most marine gas pools in South China. To improve the understanding of marine gas pools in South China with severely deformed geological background, the dominant control factors are discussed in this paper. The fluid sources, including the gas cracked from crude oil, the gas dissolved in water, the gas of inorganic origin, hydrocarbons generated during the second phase, and the mixed pool fluid source, were the most significant control factors of the types and the development stage of pools. The period of the pool formation and the reconstruction controlled the pool evolution and the distribution on a regional scale. Owing to the multiple periods of the pool formation and the reconstruction, the distribution of marine gas pools was complex both in space and in time, and the gas in the pools is heterogeneous. Pool elements, such as preservation conditions, traps and migration paths, and reservoir rocks and facies, also served as important control factors to marine gas pools in South China. Especially, the preservation conditions played a key role in maintaining marine oil and gas accumulations on a regional or local scale. According to several dominant control factors of a pool, the pool-controlling model can be constructed. As an example, the pool-controlling model of Sinian gas pool in Weiyuan gas field in Sichuan basin was summed up. ?? Higher Education Press and Springer-Verlag 2007.

  2. Cold Pool and Surface Flux Interactions in Different Environments

    Science.gov (United States)

    Grant, L. D.; van den Heever, S. C.

    2015-12-01

    Cold pools play important roles in tropical and midlatitude deep convective initiation and organization through their influence on near-surface kinematic and thermodynamic fields. Because temperature, moisture, and winds are perturbed within cold pools, cold pools can also impact surface sensible and latent heat fluxes. In turn, surface fluxes both within the cold pool and in the environment can modify the characteristics of cold pools and their evolution, with subsequent implications for convective initiation and organization. The two-way interaction between cold pools and surface energy fluxes has not been well studied and is likely to vary according to the environment and surface type. The goal of this study is therefore to investigate the mechanisms by which surface fluxes and cold pools interact in environmental conditions ranging from tropical oceanic to dry continental. This goal will be accomplished using high-resolution (grid spacings as fine as 10 m), idealized, 2D simulations of isolated cold pools; such modeling experiments have proven useful for investigating cold pools and their dynamics in many previous studies. In the proposed experiments, the surface flux formulation, surface type, and environmental conditions will be systematically varied. The impact of surface fluxes on various cold pool characteristics and their evolution, including the buoyancy, maximum vertical velocity, and moisture distribution, will be analyzed and presented. Results suggest that the mechanisms by which surface fluxes and cold pools interact vary substantially with the environment. Additionally, the indirect effects of surface fluxes on turbulent entrainment rates into the cold pool are found to play an important role in cold pool evolution. These results suggest that surface fluxes can impact the timing and manner in which cold pools initiate convection, and that their effects may be important to incorporate into cold pool parameterizations for climate simulations.

  3. Dominant factors in controlling marine gas pools in South China

    Institute of Scientific and Technical Information of China (English)

    XU Sihuang; W.Lynn Watney

    2007-01-01

    In marine strata from Sinian to Middle Triassic in South China,there develop four sets of regional and six sets of local source rocks,and ten sets of reservoir rocks.The occurrence of four main formation periods in association with five main reconstruction periods,results in a secondary origin for the most marine gas pools in South China.To improve the understanding of marine gas pools in South China with severely deformed geological background,the dominant control factors are discussed in this paper.The fluid sources,including the gas cracked from crude oil,the gas dissolved in water,the gas of inorganic origin,hydrocarbons generated during the second phase,and the mixed pool fluid source,were the most significant control factors of the types and the development stage of pools.The period of the pool formation and the reconstruction controlled the pool evolution and the distribution on a regional scale.Owing to the multiple periods of the pool formation and the reconstruction,the distribution of marine gas pools was complex both in space and in time,and the gas in the pools is heterogeneous.Pool elements,such as preservation conditions,traps and migration paths,and reservoir rocks and facies,also served as important control factors to marine gas pools in South China.Especially,the preservation conditions played a key role in maintaining marine oil and gas accumulations on a regional or local scale.According to several dominant control factors of a pool,the pool-controlling model can be constructed.As an example,the pool-controlling model of Sinian gas pool in Weiyuan gas field in Sichuan basin was summed up.

  4. An introduction to mid-Atlantic seasonal pools

    Science.gov (United States)

    Brown, L.J.; Jung, R.E.

    2005-01-01

    Seasonal pools, also known as vernal ponds, provide important ecological services to the mid-Atlantic region. This publication serves as an introduction to seasonal pool ecology and management; it also provides tools for exploring seasonal pools, including a full-color field guide to wildlife. Seasonal pools are defined as having four distinctive features: surface water isolation, periodic drying, small size and shallow depth, and support of a characteristic biological community. Seasonal pools experience regular drying that excludes populations of predatory fish. Thus, pools in the mid-Atlantic region provide critical breeding habitat for amphibian and invertebrate species (e.g., spotted salamander (Ambystoma maculatum), wood frog (Rana sylvatica), and fairy shrimp (Order Anostraca)) that would be at increased risk of predation in more permanent waters. The distinctive features of seasonal pools also make them vulnerable to human disturbance. In the mid-Atlantic region, land-use changes pose the greatest challenges to seasonal pool conservation. Seasonal pools are threatened by direct loss (e.g., filling or draining of the pool) as well as by destruction and fragmentation of adjoining terrestrial habitat. Many of the species that depend on seasonal pools for breeding spend the majority of their lives in the surrounding lands that extend a radius of 1000 feet or more from the pools; these vital habitats are being transected by roads and converted to other land uses. Other threats to seasonal pools include biological introductions and removals, mosquito control practices, amphibian diseases, atmospheric deposition, and climate change. The authors recommend a three-pronged strategy for seasonal pool conservation and management in the mid-Atlantic region: education and research, inventory and monitoring of seasonal pools, and landscape-level planning and management.

  5. Pool boiling visualization on open microchannel surfaces

    Directory of Open Access Journals (Sweden)

    Kaniowski Robert

    2017-01-01

    Full Text Available The paper presents visualization investigations into pool boiling heat transfer for open minichannel surfaces. The experiments were carried out wih saturated water at atmospheric pressure. Parallel microchannels fabricated by machining were about 0.3 mm wide and 0.2 to 0.4 mm deep. High-speed videos were used as an aid to understanding the heat transfer mechanism. The visualization study aimed at identifying nucleation sites of the departing bubbles and determining their diameters and frequency at various superheats.

  6. Subcooled pool boiling CHF in ethanol

    OpenAIRE

    Park, Jongdoc; Fukuda, Katsuya; Liu, Qiusheng

    2006-01-01

    Steady-state and transient critical heat fluxes (CHFs) were measured using a 1.0 mm diameter horizontal cylinder in a pool of highly wetting liquid, such as ethanol, due to steady and transient heat generation rate for wide range of subcoolings and pressures. Boiling CHF was assumed to happen based on a kind of hydrodynamic instability (HI) at CHF, and the model is supposed that the increase in vapor generation from the cylinder surface causes a limit of the steady-state vapor escape flow whe...

  7. Four Engineers...

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    There are four engineers traveling in a car;a mechanical engineer,a chemical engi-neer,an electrical engineer and a comput-er engineer.The car breaks down.“Sounds to me as if the pistons have seized.We ll have to strip down the engine before we canget the car working again,”says the mechanical

  8. EDITORIAL: In vitro experiments with primary mammalian cells: To Pool or not to Pool?

    Directory of Open Access Journals (Sweden)

    MJ Stoddart

    2012-11-01

    Full Text Available There are various strategies that can be adopted when performing in vitro experiments with primary cells such as mesenchymal stem cells. It is generally accepted that multiple donors need to be investigated to take into account donor to donor variability; this is especially critical when investigating primary human cells. However, increasingly it is being seen that studies are pooling the cells from multiple donors prior to performing the experiment. This has obvious advantages but also many disadvantages, the greatest being loss of statistical power. This loss of statistical power is the reason why the pooling of primary cells for experiments is not to be recommended.

  9. Weld pool vision sensing and image processing for GMAW

    Institute of Scientific and Technical Information of China (English)

    YUE Jianfeng; LI Liangyu; FAN Fanglei; WU Baolin

    2009-01-01

    It is difficult to acquire satisfied weld pool image by CCD sensor during gas metal arc welding(GMAW), for arc disturbs violently, welding current is great and working frequeacy is high. By using CMOS vision sensor to GMA W process, the vivid weld pool image is collected at any time, furthermore, whose gray compression ratio is controllable by sensor hardware circuit developed. Acquired weld pool image is firstly pre-processed by using Wiener filter and Ostu threshold segmentation algorithm. Subsequently separation between weld pool intage and cathode mist region is conducted by means of mathematical morphological algorithm, and the edge of weld pool image is extracted by using Prewitt algorithm.

  10. RECONSTRUCTION OF WELD POOL SURFACE BASED ON SHAPE FROM SHADING

    Institute of Scientific and Technical Information of China (English)

    DU Quanying; CHEN Shanben; LIN Tao

    2006-01-01

    A valid image-processing algorithm of weld pool surface reconstruction according to an input image of weld pool based on shape from shading (SFS) in computer vision is presented. The weld pool surface information is related to the backside weld width, which is crucial to the quality of weldjoint. The image of weld pool is recorded with an optical sensing method. Firstly, the reflectance map model, which specifies the imaging process, is estimated. Then, the algorithm of weld pool surface reconstruction based on SFS is implemented by iteration scheme and speeded by hierarchical structure. The results indicate the accuracy and effectiveness of the approach.

  11. THE CLIMATE FEATURES OF THE SOUTH CHINA SEA WARM POOL

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    There exists a warm pool in the South China Sea (SCS). The temporal and spatial distribution and evolution of SCS warm pool is investigated using water temperatures at a depth of 20 m in the sea. The formation of the warm pool is discussed by combining water temperatures with geostrophic currents and simulated oceanic circulation. It is found that there are significant seasonal and interannual changes in the warm pool and in association with the general circulation of the atmosphere. The development of SCS warm pool is also closely related to the gyre activities in the sea and imported warm water from Indian Ocean (Java Sea) besides radiative warming.

  12. Plant diversity associated with pools in natural and restored peatlands

    Directory of Open Access Journals (Sweden)

    N. Fontaine

    2007-06-01

    Full Text Available This study describes plant assemblages associated with the edges of peatland pools. We conducted inventories in six natural peatlands in the province of Québec (Canada in order to measure the contribution of pools to species diversity in climatic regions where peatlands are used for peat extraction. We also carried out vegetation surveys in a peatland that has been restored after peat extraction/harvesting to determine whether pool vegetation establishes along the edges of created pools when dry surface restoration techniques only are used. Pools enhanced plant species richness in natural peatlands. Around created pools, species associated with natural pools were still absent, and non-bog species were present, six years after restoration. On this basis, we emphasise the importance of preserving natural peatlands with pools. In order to restore fully the plant diversity associated with peatlands at harvested sites, it may be necessary to modify pool excavation techniques so that created pools resemble more closely those in natural peatlands. Active introduction of the plant species or communities associated with natural pools may also be needed; candidate species for North America include Andromeda glaucophylla, Cladopodiella fluitans, Carex limosa, Eriophorum virginicum, Rhynchospora alba and Sphagnum cuspidatum.

  13. Opportunities and challenges when pooling milk samples using ELISA

    DEFF Research Database (Denmark)

    Græsbøll, Kaare; Andresen, Lars Ole; Hisham Beshara Halasa, Tariq

    2017-01-01

    to standard pooling. Their efficiency and the potential savings were determined as a function of prevalence and the number of pooled samples. The potential benefit of pooling samples is dependent upon the changes in the analytical sensitivity and specificity of the test used when diluting test......-positive samples by pooling. To illustrate this, the sensitivity of antibody ELISA on pooled samples of bovine milk for Salmonella Dublin, Mycobacterium avium spp. paratuberculosis, and bovine virus diarrhea was tested. For these milk assays, the analytical sensitivity decreased rapidly with increasing pool sizes...... of samples, the two-stage non-hierarchical test may be more efficient, both in terms of number of tests and overall cost. In order to apply these results in different laboratory settings, a free Shiny WebApp was developed, to compare several pooling schemes with different cost parameters....

  14. Design of inventory pools in spare part support operation systems

    Science.gov (United States)

    Mo, Daniel Y.; Tseng, Mitchell M.; Cheung, Raymond K.

    2014-06-01

    The objective of a spare part support operation is to fulfill the part request order with different service contracts in the agreed response time. With this objective to achieve different service targets for multiple service contracts and the considerations of inventory investment, it is not only important to determine the inventory policy but also to design the structure of inventory pools and the order fulfilment strategies. In this research, we focused on two types of inventory pools: multiple inventory pool (MIP) and consolidated inventory pool (CIP). The idea of MIP is to maintain separated inventory pools based on the types of service contract, while CIP solely maintains a single inventory pool regardless of service contract. Our research aims to design the inventory pool analytically and propose reserve strategies to manage the order fulfilment risks in CIP. Mathematical models and simulation experiments would be applied for analysis and evaluation.

  15. Vision detection of weld pool shape parameters and numerical simulation

    Science.gov (United States)

    Yuan, Youzhi; Liu, Nansheng; Wang, Yanfeng; Luo, Wei; Liu, Xiaorui

    2008-12-01

    An active vision sensing system which established by low power laser and common CCD is used to shoot GTAW welding pool images. Based on choose the appropriate LASER and filters, projected angle of laser and shoot angle of CCD, we avoid the disturbance of arc effectively and obtain clear images of the welding pool. The shape parameters of welding pool can obtain by demarcated and calculated. And then a three-dimentional numerical model of GTAW welding pool is established based on the theories of fluid dynamics and heat transfer. It considered the convection heat transfer of liquid metal and heat conduction of solid metal. And FLUENT software is used for simulate the GTAW welding pool. Extract the weld pool shape parameters and done comparative study with the vision detection of pool. This will be beneficial to avoid repeat experiments and improve work efficiency.

  16. The Indo-Pacific Warm Pool: critical to world oceanography and world climate

    Science.gov (United States)

    De Deckker, Patrick

    2016-12-01

    The Indo-Pacific Warm Pool holds a unique place on the globe. It is a large area [>30 × 106 km2] that is characterised by permanent surface temperature >28 °C and is therefore called the `heat engine' of the globe. High convective clouds which can reach altitudes up to 15 km generate much latent heat in the process of convection and this area is therefore called the `steam engine' of the world. Seasonal and contrasting monsoonal activity over the region is the cause for a broad seasonal change of surface salinities, and since the area lies along the path of the Great Ocean Conveyor Belt, it is coined the `dilution' basin due to the high incidence of tropical rain and, away from the equator, tropical cyclones contribute to a significant drop in sea water salinity. Discussion about what may happen in the future of the Warm Pool under global warming is presented together with a description of the Warm Pool during the past, such as the Last Glacial Maximum when sea levels had dropped by ~125 m. A call for urgent monitoring of the IPWP area is justified on the grounds of the significance of this area for global oceanographic and climatological processes, but also because of the concerned threats to human population living there.

  17. Oligomeric rare-earth metal cluster complexes with endohedral transition metal atoms

    Energy Technology Data Exchange (ETDEWEB)

    Steinberg, Simon; Zimmermann, Sina; Brühmann, Matthias; Meyer, Eva; Rustige, Christian; Wolberg, Marike; Daub, Kathrin; Bell, Thomas; Meyer, Gerd, E-mail: gerd.meyer@uni-koeln.de

    2014-11-15

    Comproportionation reactions of rare-earth metal trihalides (RX{sub 3}) with the respective rare-earth metals (R) and transition metals (T) led to the formation of 22 oligomeric R cluster halides encapsulating T, in 19 cases for the first time. The structures of these compounds were determined by single-crystal X-ray diffraction and are composed of trimers ((T{sub 3}R{sub 11})X{sub 15}-type, P6{sub 3}/m), tetramers ((T{sub 4}R{sub 16})X{sub 28}(R{sub 4}) (P-43m), (T{sub 4}R{sub 16})X{sub 20} (P4{sub 2}/nnm), (T{sub 4}R{sub 16})X{sub 24}(RX{sub 3}){sub 4} (I4{sub 1}/a) and (T{sub 4}R{sub 16})X{sub 23} (C2/m) types of structure) and pentamers ((Ru{sub 5}La{sub 14}){sub 2}Br{sub 39}, Cc) of (TR{sub r}){sub n} (n=2–5) clusters. These oligomers are further enveloped by inner (X{sup i}) as well as outer (X{sup a}) halido ligands, which possess diverse functionalities and interconnect like oligomers through i–i, i–a and/or a–i bridges. The general features of the crystal structures for these new compounds are discussed and compared to literature entries as well as different structure types with oligomeric T centered R clusters. Dimers and tetramers originating from the aggregation of (TR{sub 6}) octahedra via common edges are more frequent than trimers and pentamers, in which the (TR{sub r}) clusters share common faces. - Graphical abstract: Rare earth-metal cluster complexes with endohedral transition metal atoms (TR{sub 6}) may connect via common edges or faces to form dimers, trimers, tetramers and pentamers of which the tetramers are the most prolific. Packing effects and electron counts play an important role. - Highlights: • Rare-earth metal cluster complexes encapsulate transition metal atoms. • Oligomers are built via connection of octahedral clusters via common edges or faces. • Dimers through pentamers with closed structures are known. • Tetramers including a tetrahedron of endohedral atoms are the most prolific.

  18. Molecular underpinnings of synaptic vesicle pool heterogeneity.

    Science.gov (United States)

    Crawford, Devon C; Kavalali, Ege T

    2015-04-01

    Neuronal communication relies on chemical synaptic transmission for information transfer and processing. Chemical neurotransmission is initiated by synaptic vesicle fusion with the presynaptic active zone resulting in release of neurotransmitters. Classical models have assumed that all synaptic vesicles within a synapse have the same potential to fuse under different functional contexts. In this model, functional differences among synaptic vesicle populations are ascribed to their spatial distribution in the synapse with respect to the active zone. Emerging evidence suggests, however, that synaptic vesicles are not a homogenous population of organelles, and they possess intrinsic molecular differences and differential interaction partners. Recent studies have reported a diverse array of synaptic molecules that selectively regulate synaptic vesicles' ability to fuse synchronously and asynchronously in response to action potentials or spontaneously irrespective of action potentials. Here we discuss these molecular mediators of vesicle pool heterogeneity that are found on the synaptic vesicle membrane, on the presynaptic plasma membrane, or within the cytosol and consider some of the functional consequences of this diversity. This emerging molecular framework presents novel avenues to probe synaptic function and uncover how synaptic vesicle pools impact neuronal signaling.

  19. Visual attention and flexible normalization pools

    Science.gov (United States)

    Schwartz, Odelia; Coen-Cagli, Ruben

    2013-01-01

    Attention to a spatial location or feature in a visual scene can modulate the responses of cortical neurons and affect perceptual biases in illusions. We add attention to a cortical model of spatial context based on a well-founded account of natural scene statistics. The cortical model amounts to a generalized form of divisive normalization, in which the surround is in the normalization pool of the center target only if they are considered statistically dependent. Here we propose that attention influences this computation by accentuating the neural unit activations at the attended location, and that the amount of attentional influence of the surround on the center thus depends on whether center and surround are deemed in the same normalization pool. The resulting form of model extends a recent divisive normalization model of attention (Reynolds & Heeger, 2009). We simulate cortical surround orientation experiments with attention and show that the flexible model is suitable for capturing additional data and makes nontrivial testable predictions. PMID:23345413

  20. Hardening Stratum, the Bitcoin Pool Mining Protocol

    Directory of Open Access Journals (Sweden)

    Recabarren Ruben

    2017-07-01

    Full Text Available Stratum, the de-facto mining communication protocol used by blockchain based cryptocurrency systems, enables miners to reliably and efficiently fetch jobs from mining pool servers. In this paper we exploit Stratum’s lack of encryption to develop passive and active attacks on Bitcoin’s mining protocol, with important implications on the privacy, security and even safety of mining equipment owners. We introduce StraTap and ISP Log attacks, that infer miner earnings if given access to miner communications, or even their logs. We develop BiteCoin, an active attack that hijacks shares submitted by miners, and their associated payouts. We build BiteCoin on WireGhost, a tool we developed to hijack and surreptitiously maintain Stratum connections. Our attacks reveal that securing Stratum through pervasive encryption is not only undesirable (due to large overheads, but also ineffective: an adversary can predict miner earnings even when given access to only packet timestamps. Instead, we devise Bedrock, a minimalistic Stratum extension that protects the privacy and security of mining participants. We introduce and leverage the mining cookie concept, a secret that each miner shares with the pool and includes in its puzzle computations, and that prevents attackers from reconstructing or hijacking the puzzles.

  1. Effects of aerobic exercise intervention on serum cartilage oligomeric matrix protein levels and lymphocyte dna damage in obese elderly females

    Science.gov (United States)

    Cho, Su Youn; Roh, Hee Tae

    2016-01-01

    [Purpose] The aim of the reported research was to investigate the effects of regular aerobic exercise on cartilage oligomeric matrix protein and oxidative DNA damage in obese, elderly females. [Subjects and Methods] Sixteen class I obese, elderly females, according to World Health Organization criteria, were randomly and equally assigned to a control group (n=8) or an exercise group (n=8). The exercise group participated in exercise sessions of 60 minutes per day, 3 days per week, for a period of 8 weeks. [Results] After aerobic exercise intervention, weight, body mass index, body fat, waist circumference, and DNA damage (Tail moment) were significantly decreased, compared with baseline values. In contrast, serum cartilage oligomeric matrix protein levels were not significantly different among any groups or time-points. [Conclusion] Regular aerobic exercise may be effective for reducing obesity-induced high DNA damage levels in obese females, without causing the deformation or degradation of lower extremity articular cartilage. PMID:27390441

  2. Binding of an Oligomeric Ellagitannin Series to Bovine Serum Albumin (BSA): Analysis by Isothermal Titration Calorimetry (ITC).

    Science.gov (United States)

    Karonen, Maarit; Oraviita, Marianne; Mueller-Harvey, Irene; Salminen, Juha-Pekka; Green, Rebecca J

    2015-12-16

    A unique series of oligomeric ellagitannins was used to study their interactions with bovine serum albumin (BSA) by isothermal titration calorimetry. Oligomeric ellagitannins, ranging from monomer to heptamer and a mixture of octamer-undecamers, were isolated as individual pure compounds. This series allowed studying the effects of oligomer size and other structural features. The monomeric to trimeric ellagitannins deviated most from the overall trends. The interactions of ellagitannin oligomers from tetramers to octa-undecamers with BSA revealed strong similarities. In contrast to the equilibrium binding constant, enthalpy showed an increasing trend from the dimer to larger oligomers. It is likely that first the macrocyclic part of the ellagitannin binds to the defined binding sites on the protein surface and then the "flexible tail" of the ellagitannin coats the protein surface. The results highlight the importance of molecular flexibility to maximize binding between the ellagitannin and protein surfaces.

  3. Characterization of oligomerization of a peptide from the ebola virus glycoprotein by small-angle neutron scattering

    Energy Technology Data Exchange (ETDEWEB)

    Egorov, V. V., E-mail: vlaegur@omrb.pnpi.spb.ru [National Research Center “Kurchatov Institute”, Konstantinov Petersburg Nuclear Physics Institute (Russian Federation); Gorshkov, A. N. [Ministry of Health of the Russian Federation, Research Institute of Influenza (Russian Federation); Murugova, T. N. [Joint Institute for Nuclear Research (Russian Federation); Vasin, A. V. [Ministry of Health of the Russian Federation, Research Institute of Influenza (Russian Federation); Lebedev, D. V.; Isaev-Ivanov, V. V. [National Research Center “Kurchatov Institute”, Konstantinov Petersburg Nuclear Physics Institute (Russian Federation); Kiselev, O. I. [Ministry of Health of the Russian Federation, Research Institute of Influenza (Russian Federation)

    2016-01-15

    Transmission electron microscopy (TEM) and small-angle neutron scattering (SANS) studies showed that model peptides QNALVCGLRQ (G33) and QNALVCGLRG (G31) corresponding to region 551–560 of the GP protein of the Sudan Ebola virus are prone to oligomerization in solution. Both peptides can form amyloid-like fibrills. The G33 peptide forms fibrils within one day of incubation, whereas the fibrillogenesis of the G31 peptide is observed only after incubation for several months. The possible role of the observed processes in the pathogenesis and the possibility of applying a combination of the TEM and SANS techniques to search for new compounds that are able to influence the protein oligomerization are discussed.

  4. Bax monomers form dimer units in the membrane that further self-assemble into multiple oligomeric species

    Science.gov (United States)

    Subburaj, Yamunadevi; Cosentino, Katia; Axmann, Markus; Pedrueza-Villalmanzo, Esteban; Hermann, Eduard; Bleicken, Stephanie; Spatz, Joachim; García-Sáez, Ana J.

    2015-08-01

    Bax is a key regulator of apoptosis that mediates the release of cytochrome c to the cytosol via oligomerization in the outer mitochondrial membrane before pore formation. However, the molecular mechanism of Bax assembly and regulation by other Bcl-2 members remains obscure. Here, by analysing the stoichiometry of Bax oligomers at the single-molecule level, we find that Bax binds to the membrane in a monomeric state and then self-assembles in Bax does not exist in a unique oligomeric state, but as several different species based on dimer units. Moreover, we show that cBid activates Bax without affecting its assembly, while Bcl-xL induces the dissociation of Bax oligomers. On the basis of our experimental data and theoretical modelling, we propose a new mechanism for the molecular pathway of Bax assembly to form the apoptotic pore.

  5. Characterization of oligomerization of a peptide from the ebola virus glycoprotein by small-angle neutron scattering

    Science.gov (United States)

    Egorov, V. V.; Gorshkov, A. N.; Murugova, T. N.; Vasin, A. V.; Lebedev, D. V.; Isaev-Ivanov, V. V.; Kiselev, O. I.

    2016-01-01

    Transmission electron microscopy (TEM) and small-angle neutron scattering (SANS) studies showed that model peptides QNALVCGLRQ (G33) and QNALVCGLRG (G31) corresponding to region 551-560 of the GP protein of the Sudan Ebola virus are prone to oligomerization in solution. Both peptides can form amyloid-like fibrills. The G33 peptide forms fibrils within one day of incubation, whereas the fibrillogenesis of the G31 peptide is observed only after incubation for several months. The possible role of the observed processes in the pathogenesis and the possibility of applying a combination of the TEM and SANS techniques to search for new compounds that are able to influence the protein oligomerization are discussed.

  6. Oligomerization and Polymerization of Ethylene Initiated by a Novel Ni(Ⅱ)-Based Acetyliminopyridine Complexes as Single-Site Catalysts

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Novel Ni(Ⅱ)-based acetyliminopyridine complexes 1b, 2b, 3b (1-3b), which are synthesized from ligands 1a, 2a, 3a (1-3a) and [NiCl2(DME)], are suitable precursors for the catalysts that are necessary for ethylene oligomerization and polymerization reactions, activated by methylaluminoxane (MAO).The MAO-treated 1-3b presents an active catalytic center, which may oligomerize and polymerize ethylene to produce linear α-olefins and polyethylene, respectively. The molecular weight distributions of oligomers that are obtained are in good agreement with the Schulz-Flory rules for oligomers>C4. The activity of oligomers show significant reliance on the structures of catalyst precursors.

  7. Synthesis, Characterization and Ethylene Oligomerization Behavior of Neutral Nickel Complexes Bearing N-Fluorinated Phenyl Salicylaldiminato Chelate Ligands

    Institute of Scientific and Technical Information of China (English)

    宋力平; 宋绍迪; 黄吉玲

    2012-01-01

    A series of neutral nickel complexes featuring N-fluorinated phenyl salicylaldiminato chelate ligands was syn- thesized, and the novel molecular structure of complex C14 was further confirmed by X-ray crystallographic analy- sis. The neutral nickel complexes showed high activity up to 9.96× 10^5 g oligomers/(mol Nioh) and high selectivity of C6 in catalyzing ethylene oligomerization using methylaluminoxane (MAO) as cocatalyst. It was observed that the strong electron-withdrawing effect of the fluorinated salicylaldiminato ligand was able to significantly increase the catalytic activity for oligomerization of ethylene. In addition, the influence of reaction parameters such as A1/Ni molar ratio, reaction temperature, a variety of cocatalyst and ethylene pressure on catalytic activity was investigated.

  8. Moyamoya disease-associated protein mysterin/RNF213 is a novel AAA+ ATPase, which dynamically changes its oligomeric state

    Science.gov (United States)

    Morito, Daisuke; Nishikawa, Kouki; Hoseki, Jun; Kitamura, Akira; Kotani, Yuri; Kiso, Kazumi; Kinjo, Masataka; Fujiyoshi, Yoshinori; Nagata, Kazuhiro

    2014-03-01

    Moyamoya disease is an idiopathic human cerebrovascular disorder that is characterized by progressive stenosis and abnormal collateral vessels. We recently identified mysterin/RNF213 as its first susceptibility gene, which encodes a 591-kDa protein containing enzymatically active P-loop ATPase and ubiquitin ligase domains and is involved in proper vascular development in zebrafish. Here we demonstrate that mysterin further contains two tandem AAA+ ATPase modules and forms huge ring-shaped oligomeric complex. AAA+ ATPases are known to generally mediate various biophysical and mechanical processes with the characteristic ring-shaped structure. Fluorescence correlation spectroscopy and biochemical evaluation suggested that mysterin dynamically changes its oligomeric forms through ATP/ADP binding and hydrolysis cycles. Thus, the moyamoya disease-associated gene product is a unique protein that functions as ubiquitin ligase and AAA+ ATPase, which possibly contributes to vascular development through mechanical processes in the cell.

  9. Increased serum cartilage oligomeric matrix protein levels and decreased patellar bone mineral density in patients with chondromalacia patellae.

    OpenAIRE

    2002-01-01

    BACKGROUND: Chondromalacia patellae is a potentially disabling disorder characterised by features of patellar cartilage degradation. OBJECTIVE: To evaluate markers of cartilage and bone turnover in patients with chondromalacia patellae. METHODS: 18 patients with chondromalacia patellae were studied. Serum cartilage oligomeric matrix protein (s-COMP) and bone sialoprotein (s-BSP) levels were measured by enzyme linked immunosorbent assay (ELISA) and compared with those of age and sex matched he...

  10. Atmospheric protein chemistry influenced by anthropogenic air pollutants: nitration and oligomerization upon exposure to ozone and nitrogen dioxide.

    Science.gov (United States)

    Liu, Fobang; Lakey, Pascale S J; Berkemeier, Thomas; Tong, Haijie; Kunert, Anna Theresa; Meusel, Hannah; Cheng, Yafang; Su, Hang; Fröhlich-Nowoisky, Janine; Lai, Senchao; Weller, Michael G; Shiraiwa, Manabu; Pöschl, Ulrich; Kampf, Christopher J

    2017-08-24

    The allergenic potential of airborne proteins may be enhanced via post-translational modification induced by air pollutants like ozone (O3) and nitrogen dioxide (NO2). The molecular mechanisms and kinetics of the chemical modifications that enhance the allergenicity of proteins, however, are still not fully understood. Here, protein tyrosine nitration and oligomerization upon simultaneous exposure of O3 and NO2 were studied in coated-wall flow-tube and bulk solution experiments under varying atmospherically relevant conditions (5-200 ppb O3, 5-200 ppb NO2, 45-96% RH), using bovine serum albumin as a model protein. Generally, more tyrosine residues were found to react via the nitration pathway than via the oligomerization pathway. Depending on reaction conditions, oligomer mass fractions and nitration degrees were in the ranges of 2.5-25% and 0.5-7%, respectively. The experimental results were well reproduced by the kinetic multilayer model of aerosol surface and bulk chemistry (KM-SUB). The extent of nitration and oligomerization strongly depends on relative humidity (RH) due to moisture-induced phase transition of proteins, highlighting the importance of cloud processing conditions for accelerated protein chemistry. Dimeric and nitrated species were major products in the liquid phase, while protein oligomerization was observed to a greater extent for the solid and semi-solid phase states of proteins. Our results show that the rate of both processes was sensitive towards ambient ozone concentration, but rather insensitive towards different NO2 levels. An increase of tropospheric ozone concentrations in the Anthropocene may thus promote pro-allergic protein modifications and contribute to the observed increase of allergies over the past decades.

  11. Protective role of Cys-178 against the inactivation and oligomerization of human insulin-degrading enzyme by oxidation and nitrosylation.

    Science.gov (United States)

    Ralat, Luis A; Ren, Min; Schilling, Alexander B; Tang, Wei-Jen

    2009-12-01

    Insulin-degrading enzyme (IDE), a 110-kDa metalloendopeptidase, hydrolyzes several physiologically relevant peptides, including insulin and amyloid-beta (Abeta). Human IDE has 13 cysteines and is inhibited by hydrogen peroxide and S-nitrosoglutathione (GSNO), donors of reactive oxygen and nitrogen species, respectively. Here, we report that the oxidative burst of BV-2 microglial cells leads to oxidation or nitrosylation of secreted IDE, leading to the reduced activity. Hydrogen peroxide and GSNO treatment of IDE reduces the V(max) for Abeta degradation, increases IDE oligomerization, and decreases IDE thermostability. Additionally, this inhibitory response of IDE is substrate-dependent, biphasic for Abeta degradation but monophasic for a shorter bradykinin-mimetic substrate. Our mutational analysis of IDE and peptide mass fingerprinting of GSNO-treated IDE using Fourier transform-ion cyclotron resonance mass spectrometer reveal a surprising interplay of Cys-178 with Cys-110 and Cys-819 for catalytic activity and with Cys-789 and Cys-966 for oligomerization. Cys-110 is near the zinc-binding catalytic center and is normally buried. The oxidation and nitrosylation of Cys-819 allow Cys-110 to be oxidized or nitrosylated, leading to complete inactivation of IDE. Cys-789 is spatially adjacent to Cys-966, and their nitrosylation and oxidation together trigger the oligomerization and inhibition of IDE. Interestingly, the Cys-178 modification buffers the inhibition caused by Cys-819 modification and prevents the oxidation or nitrosylation of Cys-110. The Cys-178 modification can also prevent the oligomerization-mediated inhibition. Thus, IDE can be intricately regulated by reactive oxygen or nitrogen species. The structure of IDE reveals the molecular basis for the long distance interactions of these cysteines and how they regulate IDE function.

  12. Screening of drugs inhibiting in vitro oligomerization of Cu/Zn-superoxide dismutase with a mutation causing amyotrophic lateral sclerosis

    OpenAIRE

    Itsuki Anzai; Keisuke Toichi; Eiichi Tokuda; Atsushi Mukaiyama; Shuji Akiyama; Yoshiaki Furukawa

    2016-01-01

    Dominant mutations in Cu/Zn-superoxide dismutase (SOD1) gene have been shown to cause a familial form of amyotrophic lateral sclerosis (SOD1-ALS). A major pathological hallmark of this disease is abnormal accumulation of mutant SOD1 oligomers in the affected spinal motor neurons. While no effective therapeutics for SOD1-ALS is currently available, SOD1 oligomerization will be a good target for developing cures of this disease. Recently, we have reproduced the formation of SOD1 oligomers ab...

  13. Synthesis, Characterization of Neutral Nickel Complexes Bearing N-Fluorophenylsalicylaldimine Chelate Ligands and Their Catalytic Activity to Ethylene Oligomerization

    Institute of Scientific and Technical Information of China (English)

    赵蔚; 黄吉玲; 朱仕正

    2005-01-01

    A series of neutral nickel complexes featuring N-fluorophenylsalicylaldimine chelate ligands was synthesized,and the molecular structure of complex 4 was further confirmed by X-ray crystallographic analysis. The neutral nickel complexes showed high activity up to 4.22×105 g oligomers/(mol Nioh) and high selectivity to C6 olefins in catalyzing ethylene oligomerization using methylaluminoxane (MAO) as cocatalyst.

  14. Transmembrane oligomeric form of Vibrio cholerae cytolysin triggers TLR2/TLR6-dependent proinflammatory responses in monocytes and macrophages.

    Science.gov (United States)

    Khilwani, Barkha; Mukhopadhaya, Arunika; Chattopadhyay, Kausik

    2015-02-15

    Vibrio cholerae cytolysin (VCC) kills target eukaryotic cells by forming transmembrane oligomeric β-barrel pores. Once irreversibly converted into the transmembrane oligomeric form, VCC acquires an unusual structural stability and loses its cytotoxic property. It is therefore possible that, on exertion of its cytotoxic activity, the oligomeric form of VCC retained in the disintegrated membrane fractions of the lysed cells would survive within the host cellular milieu for a long period, without causing any further cytotoxicity. Under such circumstances, VCC oligomers may potentially be recognized by the host immune cells. Based on such a hypothesis, in the present study we explored the interaction of the transmembrane oligomeric form of VCC with the monocytes and macrophages of the innate immune system. Our study shows that the VCC oligomers assembled in the liposome membranes elicit potent proinflammatory responses in monocytes and macrophages, via stimulation of the toll-like receptor (TLR)2/TLR6-dependent signalling cascades that involve myeloid differentiation factor 88 (MyD88)/interleukin-1-receptor-associated kinase (IRAK)1/tumour-necrosis-factor-receptor-associated factor (TRAF)6. VCC oligomer-mediated proinflammatory responses critically depend on the activation of the transcription factor nuclear factor-κB. Proinflammatory responses induced by the VCC oligomers also require activation of the mitogen-activated protein kinase (MAPK) family member c-Jun N-terminal kinase, which presumably acts via stimulation of the transcription factor activator protein-1. Notably, the role of the MAPK p38 could not be documented in the process.

  15. Both Ca2+ and Zn2+ are essential for S100A12 protein oligomerization and function

    Directory of Open Access Journals (Sweden)

    Shekhtman Alexander

    2009-04-01

    Full Text Available Abstract Background Human S100A12 is a member of the S100 family of EF-hand calcium-modulated proteins that are associated with many diseases including cancer, chronic inflammation and neurological disorders. S100A12 is an important factor in host/parasite defenses and in the inflammatory response. Like several other S100 proteins, it binds zinc and copper in addition to calcium. Mechanisms of zinc regulation have been proposed for a number of S100 proteins e.g. S100B, S100A2, S100A7, S100A8/9. The interaction of S100 proteins with their targets is strongly dependent on cellular microenvironment. Results The aim of the study was to explore the factors that influence S100A12 oligomerization and target interaction. A comprehensive series of biochemical and biophysical experiments indicated that changes in the concentration of calcium and zinc led to changes in the oligomeric state of S100A12. Surface plasmon resonance confirmed that the presence of both calcium and zinc is essential for the interaction of S100A12 with one of its extracellular targets, RAGE – the Receptor for Advanced Glycation End products. By using a single-molecule approach we have shown that the presence of zinc in tissue culture medium favors both the oligomerization of exogenous S100A12 protein and its interaction with targets on the cell surface. Conclusion We have shown that oligomerization and target recognition by S100A12 is regulated by both zinc and calcium. Our present work highlighted the potential role of calcium-binding S100 proteins in zinc metabolism and, in particular, the role of S100A12 in the cross talk between zinc and calcium in cell signaling.

  16. Increased serum cartilage oligomeric matrix protein levels and decreased patellar bone mineral density in patients with chondromalacia patellae.

    OpenAIRE

    Murphy, E; Fitzgerald, O; Saxne, Tore; Bresnihan, B

    2002-01-01

    BACKGROUND: Chondromalacia patellae is a potentially disabling disorder characterised by features of patellar cartilage degradation. OBJECTIVE: To evaluate markers of cartilage and bone turnover in patients with chondromalacia patellae. METHODS: 18 patients with chondromalacia patellae were studied. Serum cartilage oligomeric matrix protein (s-COMP) and bone sialoprotein (s-BSP) levels were measured by enzyme linked immunosorbent assay (ELISA) and compared with those of age and sex matched he...

  17. Synthesis and Catalytic Performances of a Novel Zn-MOF Catalyst Bearing Nickel Chelating Diimine Carboxylate Ligands for Ethylene Oligomerization

    Directory of Open Access Journals (Sweden)

    Suyan Liu

    2015-01-01

    Full Text Available A novel Zn-MOF[Zn3(OH2L2] was synthesized from dicarboxylate ligands with diimine groups (1,4-bis(4-CO2HC6H4-2,3-dimethyl-1,4-diazabutadiene. The physicochemical properties of the material were characterized by a series of technologies including XRD, SEM, and ICP. In order to adapt to the ethylene oligomerization process, a catalyst [Zn3OH2L1Ni2] (denoted as Cat.A possessing active Ni2+ centers was prepared by a postsynthetic treatment method using dichloride nickel as a nickel source in this work. For comparison, α-diimine ligands with/without dicarboxylic acid groups reacted with dichloride nickel to obtain homogenous Cat.B and Cat.C, respectively. The effects of reaction parameters, including n(Al/n(Ni, temperature, and pressure on the oligomerization activities and oligomers distribution were investigated. The results demonstrated that all of catalysts used with diethylaluminum chloride were active for the ethylene oligomerization. Among them, Cat.A and Cat.B showed higher catalytic activities and higher selectivities to low-carbon α-olefins at atmospheric pressure. The Cat.A exhibited the optimal catalytic activity [6.7 × 105 g/(mol·Ni·h·atm] for C4 (91.8% under the conditions of Al/Ni = 1500, P = 1.0 atm, T = 20°C. In addition, Cat.A and Cat.B presented large amount of ethylene polymer, while Cat.C had a higher catalytic activity of ethylene oligomerization at high pressure.

  18. VLAN classification and IP pool management in IaaS cloud resource pools%IaaS云资源池中VLAN划分及IP池管理*

    Institute of Scientific and Technical Information of China (English)

    李小康; 廖建新; 沈奇威; 曹予飞

    2013-01-01

    VLAN划分和IP池管理是IaaS云资源池对网络资源进行分配的重要环节,合理规划管理VLAN和IP地址对于提高网络安全性、稳定性、高效性有非常重要的意义。本文从工程实践角度,将传统的VLAN及IP地址分配方式与云计算资源池网络自动化的特点相结合,进行实际应用方面的设计,有着重要的实践意义。%VLAN classiifcation and IP pool management are important parts of network resources allocation in IaaS cloud resource pool management. It has a very important signiifcance in the improvement of network security, stability, efficiency to appropriately manage VLAN and IP address. From the perspective of engineering practice, this article combines the traditional VLAN and IP address allocation with the characteristics of cloud computing resource pools network automation to realize the practical application, thus has important practical signiifcance.

  19. Expanding the live kidney donor pool: ethical considerations regarding altruistic donors, paired and pooled programs.

    Science.gov (United States)

    Patel, Shaneel Rajendra; Chadha, Priyanka; Papalois, Vassilios

    2011-06-01

    In renal transplant, there is a well-known deficiency in organ supply relative to demand. Live donation provides superior results when compared with deceased donation including a better rate of graft success and fewer immunologic complications. This deficiency in organs leads to significant morbidity and mortality rates. Alternative avenues have been extensively explored that may expand the live donor pool. They include altruistic donation as well as paired and pooled exchange programs. Altruistic donation is a truly selfless act from a donor unknown to the recipient. Kidney paired donation involves 2 incompatible donor-recipient pairs swapping donors to produce compatibility. Pooled donation involves at least 2 pairs, and can take the form of domino chains in which altruistic input sets up a chain of transplants, in which each recipient's incompatible donor makes a donation for the next recipient. Despite application of these various methods, there lie extensive ethical issues surrounding them. Misconceptions frequently occur; for instance, the perceived benefit that donating an organ to a loved one is greater for a related donor than for an altruistic one. Additionally, it is frequently believed that immunologic incompatibility offers coerced donors liberation from surgery, and that overcoming these barriers by introducing exchange programs provides vulnerable donors less protection. This article explores these and other complex ethical issues surrounding the various methods of expanding the donor pool. The authors offer opinions that challenge the ethical issues and attempt to overcome those views that hinder progress in the field.

  20. Oligomeric adiponectin forms and their complexes in the blood of healthy donors and patients with type 2 diabetes mellitus.

    Science.gov (United States)

    Kogan, Alexander E; Filatov, Vladimir L; Kolosova, Olga V; Katrukha, Ivan A; Mironova, Ekaterina V; Zhuravleva, Natalya S; Nagibin, Oleg A; Kara, Andrei N; Bereznikova, Anastasiya V; Katrukha, Alexey G

    2013-01-01

    Adiponectin (Adn) is a protein that circulates in the blood in several oligomeric forms, namely low-, medium-, and high-molecular-weight forms. Adn may serve as a risk factor for type 2 diabetes mellitus (T2DM). The aims of this work were (1) to produce monoclonal antibodies (MAbs) specific to different Adn oligomeric forms, (2) to design immunoassays suitable for measuring the Adn forms present in human blood, and (3) to investigate the changes in Adn forms that occur in patients with T2DM. Gel filtration, fluoroimmunoassays, and Western blotting were utilized as major techniques in this study. MAbs recognizing various oligomeric forms of Adn were obtained. Complexes between Adn and complement component C1q and between the low molecular weight form of Adn and albumin were described in human blood. A decrease in the total Adn and Adn-albumin complex levels in the blood of patients with T2DM and no difference in the levels of the Adn-C1q complex in comparison with healthy volunteers were demonstrated. An Adn94-Adn63 fluoroimmunoassay was selected as the technique that most accurately measured the mass ratio of Adn oligomers in blood samples, and an Adn214-Adn27 assay that measured the low-molecular-weight form of Adn only.

  1. Understanding Peptide Oligomeric State in Langmuir Monolayers of Amphiphilic 3-Helix Bundle-Forming Peptide-PEG Conjugates

    Science.gov (United States)

    Shu, Jessica Y.; Xu, Ting

    2016-01-01

    Coiled-coil peptide–polymer conjugates are an emerging class of biomaterials. Fundamental understanding of the coiled-coil oligomeric state and assembly process of these hybrid building blocks is necessary to exert control over their assembly into well-defined structures. Here, we studied the effect of peptide structure and PEGylation on the self-assembly process and oligomeric state of a Langmuir monolayer of amphiphilic coiled-coil peptide–polymer conjugates using X-ray reflectivity (XR) and grazing-incidence X-ray diffraction (GIXD). Our results show that the oligomeric state of PEGylated amphiphiles based on 3-helix bundle-forming peptide is surface pressure dependent, a mixture of dimers and trimers was formed at intermediate surface pressure but transitions into trimers completely upon increasing surface pressure. Moreover, the interhelical distance within the coiled-coil bundle of 3-helix peptide-PEG conjugate amphiphiles was not perturbed under high surface pressure. Present studies provide valuable insights into the self-assembly process of hybrid peptide–polymer conjugates and guidance to develop biomaterials with controlled multivalency of ligand presentation. PMID:27784156

  2. Metal-dependent SpoIIE oligomerization stabilizes FtsZ during asymmetric division in Bacillus subtilis

    Science.gov (United States)

    Król, Ewa; de Sousa Borges, Anabela; Kopacz, Malgorzata

    2017-01-01

    SpoIIE is a bifunctional protein involved in asymmetric septum formation and in activation of the forespore compartment-specific transcription factor σF through dephosphorylation of SpoIIAA-P. The phosphatase activity of SpoIIE requires Mn2+ as a metal cofactor. Here, we show that the presence of a metal cofactor also influences SpoIIE oligomerization and asymmetric septum formation. Absence of Mn2+ from sporulation medium results in a delay of the formation of polar FtsZ-rings, similar to a spoIIE null mutant. We purified the entire cytoplasmic part of the SpoIIE protein, and show that the protein copurifies with bound metals. Metal binding both stimulates SpoIIE oligomerization, and results in the formation of larger oligomeric structures. The presence of SpoIIE oligomers reduces FtsZ GTP hydrolysis activity and stabilizes FtsZ polymers in a light scattering assay. Combined, these results indicate that metal binding is not just required for SpoIIE phosphatase activity but also is important for SpoIIE's role in asymmetric septum formation. PMID:28358838

  3. Irradiation of Microbes from Spent Nuclear Fuel Storage Pool Environments

    Energy Technology Data Exchange (ETDEWEB)

    Breckenridge, C.R.; Watkins, C.S.; Bruhn, D.F.; Roberto, F.F.; Tsang, M.N.; Pinhero, P.J. [INEEL (US); Brey, R.F. [ISU (US); Wright, R.N.; Windes, W.F.

    1999-09-03

    Microbes have been isolated and identified from spent nuclear fuel storage pools at the Idaho National Engineering and Environmental Laboratory (INEEL). Included among these are Corynebacterium aquaticum, Pseudomonas putida, Comamonas acidovorans, Gluconobacter cerinus, Micrococcus diversus, Rhodococcus rhodochrous, and two strains of sulfate-reducing bacteria (SRB). We examined the sensitivity of these microbes to a variety of total exposures of radiation generated by a 6-MeV linear accelerator (LINAC). The advantage of using a LINAC is that it provides a relatively quick screen of radiation tolerance. In the first set of experiments, we exposed each of the aforementioned microbes along with four additional microbes, pseudomonas aeruginosa, Micrococcus luteus, Escherchia coli, and Deinococcus radiodurans to exposures of 5 x 10{sup 3} and 6 x 10{sup 4} rad. All microbial specimens withstood the lower exposure with little or no reduction in cell population. Upon exposing the microbes to the larger dose of 6 x 10{sup 4} rad, we observed two distinct groupings: microbes that demonstrate resistance to radiation, and microbes that display intolerance through a dramatic reduction from their initial population. Microbes in the radiation tolerant grouping were exposed to 1.1 x 10{sup 5} rad to examine the extent of their resistance. We observe a correlation between radiation resistance and gram stain. The gram-positive species we examined seem to demonstrate a greater radiation resistance.

  4. To pool or not to pool? Impact of the use of individual and pooled fecal samples for in vitro fermentation studies

    NARCIS (Netherlands)

    Aguirre, M.; Ramiro Garcia, J.; Koenen, M.E.; Venema, K.

    2014-01-01

    This study investigated the stability and the activity of the microbiota from a single and a pool of donors in the TNO in vitro model of the colon (TIM-2 system). Our findings demonstrate the suitability of the preparation of a pool of fecal sample to be used for fermentation experiments.

  5. To pool or not to pool? Impact of the use of individual and pooled fecal samples for in vitro fermentation studies

    NARCIS (Netherlands)

    Aguirre, M.; Ramiro-Garcia, J.; Koenen, M.E.; Venema, K.

    2014-01-01

    This study investigated the stability and the activity of the microbiota from a single and a pool of donors in the TNO in vitro model of the colon (TIM-2 system). Our findings demonstrate the suitability of the preparation of a pool of fecal sample to be used for fermentation experiments. Chemicals/

  6. Structure-properties relationships of polyhedral oligomeric silsesquioxane (POSS filled PS nanocomposites

    Directory of Open Access Journals (Sweden)

    J. J. Schwab

    2012-07-01

    Full Text Available The polyhedral oligomeric silsesquioxane (POSS additivated polystyrene (PS based nanocomposites were prepared by melt processing and the structure-properties relationships of the POSS-PS systems were compared to those of the neat PS. In order to investigate the effect of these structural parameters on the final properties of the polymer nanocomposites, five different kinds of POSS samples were used, in particular, POSS with different inorganic cage and with different organic pendent groups. The rheological investigation suggests clearly that the POSS acts as a plasticizer and that the processability of the PS was positively modified. The affinity between the POSS samples and the PS matrix was estimated by the calculated theoretical solubility parameters, considering the Hoy’s method and by morphology analysis. Minor difference between the solubility parameter of POSS and the matrix means better compatibility and no aggregation tendency. Furthermore, the POSS loading leads to a decrease of the rigidity, of the glass transition temperature and of the damping factor of the nanocomposite systems. The loading of different POSS molecules with open cage leads to a more pronounced effect on all the investigated properties that the loading of the POSS molecules with closed cage. Moreover, the melt properties are significantly influenced by the type of inorganic framework, by the type of the pendent organic groups and by the interaction between the POSS organic groups and the host matrix, while, the solid state properties appears to be influenced more by the kind of cage.

  7. Microwave dielectric study of an oligomeric electrolyte gelator by time domain reflectometry.

    Science.gov (United States)

    Kundu, Shyamal Kumar; Yagihara, Shin; Yoshida, Masaru; Shibayama, Mitsuhiro

    2009-07-30

    The dynamics of water molecules in aqueous solutions of an oligomeric electrolyte gelator, poly[pyridinium-1,4-diyliminocarbonyl-1,4-phenylene-methylene chloride] (1-Cl) was characterized by microwave dielectric measurements using the time domain reflectometry method. The dielectric dispersion and absorption curves related to the orientational motion of water molecules were described by the Cole-Cole equation. Discontinuities were observed in the concentration dependence of the dielectric relaxation strength, Deltaepsilonh, as well as in the Cole-Cole parameter, betah. These discontinuities were observed between the samples with concentrations of 6 and 7 g/L 1-Cl/water, which correspond to a change in the transparency. Such a discontinuity corresponds to the observation of the critical concentration of gelation. The interaction between water and 1-Cl molecules was discussed from the tauh-betah diagram. As 1-Cl carries an amide group, it could be expected that 1-Cl may interact hydrophilically with water, but the present result suggests that 1-Cl interact hydrophobically with water.

  8. Design of a minimal protein oligomerization domain by a structural approach.

    Science.gov (United States)

    Burkhard, P; Meier, M; Lustig, A

    2000-12-01

    Because of the simplicity and regularity of the alpha-helical coiled coil relative to other structural motifs, it can be conveniently used to clarify the molecular interactions responsible for protein folding and stability. Here we describe the de novo design and characterization of a two heptad-repeat peptide stabilized by a complex network of inter- and intrahelical salt bridges. Circular dichroism spectroscopy and analytical ultracentrifugation show that this peptide is highly alpha-helical and 100% dimeric tinder physiological buffer conditions. Interestingly, the peptide was shown to switch its oligomerization state from a dimer to a trimer upon increasing ionic strength. The correctness of the rational design principles used here is supported by details of the atomic structure of the peptide deduced from X-ray crystallography. The structure of the peptide shows that it is not a molten globule but assumes a unique, native-like conformation. This de novo peptide thus represents an attractive model system for the design of a molecular recognition system.

  9. Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase.

    Science.gov (United States)

    Wu, Zhengliang L; Zhou, Hui; Ethen, Cheryl M; N Reinhold, Vernon

    2016-04-29

    The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. Yet to date, the structure component responsible for its virulence is still a mystery. In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. In this report, the N-glycans from both forms were released and characterized by mass spectrometry. It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. These findings were further supported by the results obtained from in vitro incorporation of azido fucose and (3)H-labeled fucose using core-6 fucosyltransferase, FUT8. In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme.

  10. Oligomeric synthesis and density functional theory of leucoemeraldine base form of polyaniline

    Science.gov (United States)

    Ullah, Rizwan; Ullah, Habib; Shah, Anwar-ul-Haq Ali; Bilal, Salma; Ali, Khurshid

    2017-01-01

    Oligomeric synthesis of phenyl-end-capped oligoaniline (4PANI LB) has been carried out through a weak oxidizing agent, CuCl2, using chemical oxidative polymerization protocol. The sample was characterized by mass spectrometry, UV-vis, IR, and CHN elemental analysis. The experimental results are counterchecked with the aid of Quantum mechanical calculations such as density functional theory (DFT). DFT at B3LYP/6-31 G (d) level of theory was used for the geometric and electronic properties simulations which also confirm the existence of 4PANI LB. Excellent correlation is observed between the experiment and theory, particularly in the UV-vis spectra which conclude the formation of tetramer (fully reduced form) 4PANI LB (C24H20.06N4.07). Electronic properties such as Ionization Potential (I.P), Electron Affinities (E.A), the coefficient of highest occupied molecular orbital (HOMO), the coefficient of lowest unoccupied molecular orbital (LUMO) of 4PANI LB were evaluated at the above-mentioned level of theory.

  11. Determination of GLUT1 Oligomerization Parameters using Bioluminescent Förster Resonance Energy Transfer.

    Science.gov (United States)

    Looyenga, Brendan; VanOpstall, Calvin; Lee, Zion; Bell, Jed; Lodge, Evans; Wrobel, Katherine; Arnoys, Eric; Louters, Larry

    2016-06-30

    The facilitated glucose transporter GLUT1 (SLC2A1) is an important mediator of glucose homeostasis in humans. Though it is found in most cell types to some extent, the level of GLUT1 expression across different cell types can vary dramatically. Prior studies in erythrocytes-which express particularly high levels of GLUT1-have suggested that GLUT1 is able to form tetrameric complexes with enhanced transport activity. Whether dynamic aggregation of GLUT1 also occurs in cell types with more modest expression of GLUT1, however, is unclear. To address this question, we developed a genetically encoded bioluminescent Förster resonance energy transfer (BRET) assay using the luminescent donor Nanoluciferase and fluorescent acceptor mCherry. By tethering these proteins to the N-terminus of GLUT1 and performing saturation BRET analysis, we were able to demonstrate the formation of multimeric complexes in live cells. Parallel use of flow cytometry and immunoblotting further enabled us to estimate the density of GLUT1 proteins required for spontaneous oligomerization. These data provide new insights into the physiological relevance of GLUT1 multimerization as well as a new variant of BRET assay that is useful for measuring the interactions among other cell membrane proteins in live cells.

  12. Oncogenic Mutations Differentially Affect Bax Monomer, Dimer, and Oligomeric Pore Formation in the Membrane

    Science.gov (United States)

    Zhang, Mingzhen; Zheng, Jie; Nussinov, Ruth; Ma, Buyong

    2016-09-01

    Dysfunction of Bax, a pro-apoptotic regulator of cellular metabolism is implicated in neurodegenerative diseases and cancer. We have constructed the first atomistic models of the Bax oligomeric pore consisting with experimental residue-residue distances. The models are stable, capturing well double electron-electron resonance (DEER) spectroscopy measurements and provide structural details in line with the DEER data. Comparison with the latest experimental results revealed that our models agree well with both Bax and Bak pores, pointed to a converged structural arrangement for Bax and Bak pore formation. Using multi-scale molecular dynamics simulations, we probed mutational effects on Bax transformation from monomer → dimer → membrane pore formation at atomic resolution. We observe that two cancer-related mutations, G40E and S118I, allosterically destabilize the monomer and stabilize an off-pathway swapped dimer, preventing productive pore formation. This observation suggests a mechanism whereby the mutations may work mainly by over-stabilizing the monomer → dimer transformation toward an unproductive off-pathway swapped-dimer state. Our observations point to misfolded Bax states, shedding light on the molecular mechanism of Bax mutation-elicited cancer. Most importantly, the structure of the Bax pore facilitates future study of releases cytochrome C in atomic detail.

  13. Structure–property relationships in hybrid dental nanocomposite resins containing monofunctional and multifunctional polyhedral oligomeric silsesquioxanes

    Science.gov (United States)

    Wang, Weiguo; Sun, Xiang; Huang, Li; Gao, Yu; Ban, Jinghao; Shen, Lijuan; Chen, Jihua

    2014-01-01

    Organic-inorganic hybrid materials, such as polyhedral oligomeric silsesquioxanes (POSS), have the potential to improve the mechanical properties of the methacrylate-based composites and resins used in dentistry. In this article, nanocomposites of methacryl isobutyl POSS (MI-POSS [bears only one methacrylate functional group]) and methacryl POSS (MA-POSS [bears eight methacrylate functional groups]) were investigated to determine the effect of structures on the properties of dental resin. The structures of the POSS-containing networks were determined by scanning electron microscopy, transmission electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. Monofunctional POSS showed a strong tendency toward aggregation and crystallization, while multifunctional POSS showed higher miscibility with the dimethacrylate monomer. The mechanical properties and wear resistance decreased with increasing amounts of MI-POSS, indicating that the MI-POSS agglomerates act as the mechanical weak point in the dental resins. The addition of small amounts of MA-POSS improved the mechanical and shrinkage properties. However, samples with a higher MA-POSS concentration showed lower flexural strength and flexural modulus, indicating that there is a limited range in which the reinforcement properties of MA-POSS can operate. This concentration dependence is attributed to phase separation at higher concentrations of POSS, which affects the structural integrity, and thus, the mechanical and shrinkage properties of the dental resin. Our results show that resin with 3% MA-POSS is a potential candidate for resin-based dental materials. PMID:24550674

  14. Structure-property relationships in hybrid dental nanocomposite resins containing monofunctional and multifunctional polyhedral oligomeric silsesquioxanes.

    Science.gov (United States)

    Wang, Weiguo; Sun, Xiang; Huang, Li; Gao, Yu; Ban, Jinghao; Shen, Lijuan; Chen, Jihua

    2014-01-01

    Organic-inorganic hybrid materials, such as polyhedral oligomeric silsesquioxanes (POSS), have the potential to improve the mechanical properties of the methacrylate-based composites and resins used in dentistry. In this article, nanocomposites of methacryl isobutyl POSS (MI-POSS [bears only one methacrylate functional group]) and methacryl POSS (MA-POSS [bears eight methacrylate functional groups]) were investigated to determine the effect of structures on the properties of dental resin. The structures of the POSS-containing networks were determined by scanning electron microscopy, transmission electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. Monofunctional POSS showed a strong tendency toward aggregation and crystallization, while multifunctional POSS showed higher miscibility with the dimethacrylate monomer. The mechanical properties and wear resistance decreased with increasing amounts of MI-POSS, indicating that the MI-POSS agglomerates act as the mechanical weak point in the dental resins. The addition of small amounts of MA-POSS improved the mechanical and shrinkage properties. However, samples with a higher MA-POSS concentration showed lower flexural strength and flexural modulus, indicating that there is a limited range in which the reinforcement properties of MA-POSS can operate. This concentration dependence is attributed to phase separation at higher concentrations of POSS, which affects the structural integrity, and thus, the mechanical and shrinkage properties of the dental resin. Our results show that resin with 3% MA-POSS is a potential candidate for resin-based dental materials.

  15. Role of redox environment on the oligomerization of higher molecular weight adiponectin

    Directory of Open Access Journals (Sweden)

    Nuñez Martha

    2011-05-01

    Full Text Available Abstract Background Adiponectin is an adipocyte-secreted hormone with insulin-sensitizing and anti-inflammatory actions. The assembly of trimeric, hexameric, and higher molecular weight (HMW species of adiponectin is a topic of significant interest because physiological actions of adiponectin are oligomer-specific. In addition, adiponectin assembly is an example of oxidative oligomerization of multi-subunit protein complexes in endoplasmic reticulum (ER. Results We previously reported that trimers assemble into HMW adiponectin via intermediates stabilized by disulfide bonds, and complete oxidation of available cysteines locks adiponectin in hexameric conformation. In this study, we examined the effects of redox environment on the rate of oligomer formation and the distribution of oligomers. Reassembly of adiponectin under oxidizing conditions accelerated disulfide bonding but favored formation of hexamers over the HMW species. Increased ratios of HMW to hexameric adiponectin could be achieved rapidly under oxidizing conditions by promoting disulfide rearrangement. Conclusions Based upon these observations, we propose oxidative assembly of multi-subunit adiponectin complexes in a defined and stable redox environment is favored under oxidizing conditions coupled with high rates of disulfide rearrangement.

  16. DHPC strongly affects the structure and oligomerization propensity of Alzheimer's Aβ(1-40) peptide.

    Science.gov (United States)

    Dahse, Kirsten; Garvey, Megan; Kovermann, Michael; Vogel, Alexander; Balbach, Jochen; Fändrich, Marcus; Fahr, Alfred

    2010-11-05

    Alzheimer's disease (AD) is thought to depend on the deleterious action of amyloid fibrils or oligomers derived from β-amyloid (Aβ) peptide. Out of various known Aβ alloforms, the 40-residue peptide Aβ(1-40) occurs at highest concentrations inside the brains of AD patients. Its aggregation properties critically depend on lipids, and it was thus proposed that lipids could play a major role in AD. To better understand their possible effects on the structure of Aβ and on the ability of this peptide to form potentially detrimental amyloid structures, we here analyze the interactions between Aβ(1-40) and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC). DHPC has served, due to its controlled properties, as a major model system for studying general lipid properties. Here, we show that DHPC concentrations of 8 mM or higher exert dramatic effects on the conformation of soluble Aβ(1-40) peptide and induce the formation of β-sheet structure at high levels. By contrast, we find that DHPC concentrations well below the critical micelle concentration present no discernible effect on the conformation of soluble Aβ, although they substantially affect the peptide's oligomerization and fibrillation kinetics. These data imply that subtle lipid-peptide interactions suffice in controlling the overall aggregation properties and drastically accelerate, or delay, the fibrillation kinetics of Aβ peptide in near-physiological buffer solutions.

  17. An integrative approach to the study of filamentous oligomeric assemblies, with application to RecA.

    Science.gov (United States)

    Boyer, Benjamin; Ezelin, Johann; Poulain, Pierre; Saladin, Adrien; Zacharias, Martin; Robert, Charles H; Prévost, Chantal

    2015-01-01

    Oligomeric macromolecules in the cell self-organize into a wide variety of geometrical motifs such as helices, rings or linear filaments. The recombinase proteins involved in homologous recombination present many such assembly motifs. Here, we examine in particular the polymorphic characteristics of RecA, the most studied member of the recombinase family, using an integrative approach that relates local modes of monomer/monomer association to the global architecture of their screw-type organization. In our approach, local modes of association are sampled via docking or Monte Carlo simulations. This enables shedding new light on fiber morphologies that may be adopted by the RecA protein. Two distinct RecA helical morphologies, the so-called "extended" and "compressed" forms, are known to play a role in homologous recombination. We investigate the variability within each form in terms of helical parameters and steric accessibility. We also address possible helical discontinuities in RecA filaments due to multiple monomer-monomer association modes. By relating local interface organization to global filament morphology, the strategies developed here to study RecA self-assembly are particularly well suited to other DNA-binding proteins and to filamentous protein assemblies in general.

  18. Oligomeric procyanidins of French maritime pine bark extract (Pycnogenol) effectively inhibit alpha-glucosidase.

    Science.gov (United States)

    Schäfer, Angelika; Högger, Petra

    2007-07-01

    The standardized maritime pine bark extract (Pycnogenol) was reported to exert clinical anti-diabetic effects after peroral intake. However, an increased insulin secretion was not observed after administration of the extract to patients. Our aim was to elucidate whether the described clinical effects of Pycnogenol are related to inhibition of alpha-glucosidase. Therefore, we analyzed the inhibitory activity of Pycnogenol, green tea extract and acarbose towards alpha-glucosidase. Furthermore, we explored different fractions of Pycnogenol containing compounds of diverse molecular masses from polyphenolic monomers, dimers and higher oligomers to uncover which components exhibited the most pronounced inhibitory activity. We found that Pycnogenol exhibited the most potent inhibition (IC(50) about 5 microg/mL) on alpha-glucosidase compared to green tea extract (IC(50) about 20 microg/mL) and acarbose (IC(50) about 1mg/mL). The inhibitory action of Pycnogenol was stronger in extract fractions containing higher procyanidin oligomers. The results obtained assign a novel, local effect to oligomeric procyanidins and contribute to the explanation of glucose-lowering effects of Pycnogenol observed in clinical trials with diabetic patients.

  19. Oligomerization of 10,16-Dihydroxyhexadecanoic Acid and Methyl 10,16-Dihydroxyhexadecanoate Catalyzed by Lipases

    Directory of Open Access Journals (Sweden)

    Daniel Arrieta-Baez

    2013-08-01

    Full Text Available The main monomer of tomato cuticle, 10,16-dihydroxyhexadecanoic acid (10,16-DHPA and its methyl ester derivative (methyl-10,16-dihydroxyhexadecanote; methyl-10,16-DHHD, were used to study their oligomerization reactions catalyzed by five lipases: Candida antarctica lipase B (CAL-B, Rhizomucor miehei lipase (RM, Thermomyces lanuginosus lipase (TL, Pseudomonas cepacia lipase (PCL and porcine pancreatic lipase (PPL. For 10,16-DHPA, optimum yields were obtained at 60 °C using toluene and 2-methyl-2-butanol (2M2B as solvent, while for methyl-10,16-DHHD the bests yields were obtained in toluene and acetonitrile. Both reactions leaded to linear polyesters according to the NMR and FT-IR analysis, and there was no data indicating the presence of branched polymers. Using optimized conditions, poly(10,16-DHPA and poly(methyl-10,16-DHHD with Mw = 814 and Mn = 1,206 Da, and Mw = 982 and Mn = 860 Da, respectively, were formed according to their MALDI-TOF MS and ESI-MS data. The self-assembly of the polyesters obtained were analyzed by AFM.

  20. Strong Screening Effect of Polyhedral Oligomeric Silsesquioxanes (POSS Nanoparticles on Hydrogen Bonded Polymer Blends

    Directory of Open Access Journals (Sweden)

    Chin-Wei Chiou

    2014-03-01

    Full Text Available In this study we used anionic living polymerization to prepare two different homopolymers: a poly(methyl methacrylate (PMMA and a PMMA derivative presenting polyhedral oligomeric silsesquioxane (PMA-POSS units as its side chains. We then employed differential scanning calorimetry (DSC, Fourier transform infrared (FTIR spectroscopy, and wide-angle X-ray diffraction (WAXD to investigate the miscibility and specific interactions of PMMA and PMA-POSS with three hydrogen bonding donor compounds: poly(vinyl phenol (PVPh, phenolic resin, and bisphenol A (BPA. DSC revealed that all of the PVPh/PMMA, phenolic/PMMA, and BPA/PMMA blends exhibited a single glass transition temperature, characteristic of miscible systems; FTIR spectroscopic analyses revealed that such miscibility resulted from hydrogen bonding interactions between the C=O groups of PMMA and the OH groups of these three hydrogen bonding donor compounds. In contrast, all of the PVPh/PMA-POSS, phenolic/PMA-POSS, and BPA/PMA-POSS blends were immiscible: DSC revealed two glass transition temperatures arising from strong screening effects (FTIR spectroscopy and high degrees of aggregation (WAXD of the POSS nanoparticles. We propose that the value of the intramolecular screening effect (γ should be very close to 1 for all PMA-POSS blend systems when POSS nanoparticles appear as the side chains of PMMA, such that the OH groups of the hydrogen bonding donor compounds cannot interact with the C=O groups of PMA-POSS.