WorldWideScience

Sample records for decentralized mass exchange

  1. On the efficiency of decentralized exchange with resale possibilities

    DEFF Research Database (Denmark)

    Groes, Ebbe; Tranæs, Torben

    1999-01-01

    . If resale is possible and transaction costs are negligible, we would nevertheless expect an efficient allocation to result from decentralized exchange. This paper suggests that this depends on the nature of the commodity; while the allocation of a durable good will be efficient, the allocation...

  2. Heat and mass exchanger

    Science.gov (United States)

    Lowenstein, Andrew; Sibilia, Marc J.; Miller, Jeffrey A.; Tonon, Thomas

    2007-09-18

    A mass and heat exchanger includes at least one first substrate with a surface for supporting a continuous flow of a liquid thereon that either absorbs, desorbs, evaporates or condenses one or more gaseous species from or to a surrounding gas; and at least one second substrate operatively associated with the first substrate. The second substrate includes a surface for supporting the continuous flow of the liquid thereon and is adapted to carry a heat exchange fluid therethrough, wherein heat transfer occurs between the liquid and the heat exchange fluid.

  3. Analytical aspects of hydrogen exchange mass spectrometry

    Science.gov (United States)

    Engen, John R.; Wales, Thomas E.

    2016-01-01

    The analytical aspects of measuring hydrogen exchange by mass spectrometry are reviewed. The nature of analytical selectivity in hydrogen exchange is described followed by review of the analytical tools required to accomplish fragmentation, separation, and the mass spectrometry measurements under restrictive exchange quench conditions. In contrast to analytical quantitation that relies on measurements of peak intensity or area, quantitation in hydrogen exchange mass spectrometry depends on measuring a mass change with respect to an undeuterated or deuterated control, resulting in a value between zero and the maximum amount of deuterium that could be incorporated. Reliable quantitation is a function of experimental fidelity and to achieve high measurement reproducibility, a large number of experimental variables must be controlled during sample preparation and analysis. The method also reports on important qualitative aspects of the sample, including conformational heterogeneity and population dynamics. PMID:26048552

  4. Conjugate heat and mass transfer in heat mass exchanger ducts

    CERN Document Server

    Zhang, Li-Zhi

    2013-01-01

    Conjugate Heat and Mass Transfer in Heat Mass Exchanger Ducts bridges the gap between fundamentals and recent discoveries, making it a valuable tool for anyone looking to expand their knowledge of heat exchangers. The first book on the market to cover conjugate heat and mass transfer in heat exchangers, author Li-Zhi Zhang goes beyond the basics to cover recent advancements in equipment for energy use and environmental control (such as heat and moisture recovery ventilators, hollow fiber membrane modules for humidification/dehumidification, membrane modules for air purification, desi

  5. Hydrogen/deuterium exchange in mass spectrometry.

    Science.gov (United States)

    Kostyukevich, Yury; Acter, Thamina; Zherebker, Alexander; Ahmed, Arif; Kim, Sunghwan; Nikolaev, Eugene

    2018-03-30

    The isotopic exchange approach is in use since the first observation of such reactions in 1933 by Lewis. This approach allows the investigation of the pathways of chemical and biochemical reactions, determination of structure, composition, and conformation of molecules. Mass spectrometry has now become one of the most important analytical tools for the monitoring of the isotopic exchange reactions. Investigation of conformational dynamics of proteins, quantitative measurements, obtaining chemical, and structural information about individual compounds of the complex natural mixtures are mainly based on the use of isotope exchange in combination with high resolution mass spectrometry. The most important reaction is the Hydrogen/Deuterium exchange, which is mainly performed in the solution. Recently we have developed the approach allowing performing of the Hydrogen/Deuterium reaction on-line directly in the ionization source under atmospheric pressure. Such approach simplifies the sample preparation and can accelerate the exchange reaction so that certain hydrogens that are considered as non-labile will also participate in the exchange. The use of in-ionization source H/D exchange in modern mass spectrometry for structural elucidation of molecules serves as the basic theme in this review. We will focus on the mechanisms of the isotopic exchange reactions and on the application of in-ESI, in-APCI, and in-APPI source Hydrogen/Deuterium exchange for the investigation of petroleum, natural organic matter, oligosaccharides, and proteins including protein-protein complexes. The simple scenario for adaptation of H/D exchange reactions into mass spectrometric method is also highlighted along with a couple of examples collected from previous studies. © 2018 Wiley Periodicals, Inc.

  6. Life-Cycle Cost and Environmental Assessment of Decentralized Nitrogen Recovery Using Ion Exchange from Source-Separated Urine through Spatial Modeling.

    Science.gov (United States)

    Kavvada, Olga; Tarpeh, William A; Horvath, Arpad; Nelson, Kara L

    2017-11-07

    Nitrogen standards for discharge of wastewater effluent into aquatic bodies are becoming more stringent, requiring some treatment plants to reduce effluent nitrogen concentrations. This study aimed to assess, from a life-cycle perspective, an innovative decentralized approach to nitrogen recovery: ion exchange of source-separated urine. We modeled an approach in which nitrogen from urine at individual buildings is sorbed onto resins, then transported by truck to regeneration and fertilizer production facilities. To provide insight into impacts from transportation, we enhanced the traditional economic and environmental assessment approach by combining spatial analysis, system-scale evaluation, and detailed last-mile logistics modeling using the city of San Francisco as an illustrative case study. The major contributor to energy intensity and greenhouse gas (GHG) emissions was the production of sulfuric acid to regenerate resins, rather than transportation. Energy and GHG emissions were not significantly sensitive to the number of regeneration facilities. Cost, however, increased with decentralization as rental costs per unit area are higher for smaller areas. The metrics assessed (unit energy, GHG emissions, and cost) were not significantly influenced by facility location in this high-density urban area. We determined that this decentralized approach has lower cost, unit energy, and GHG emissions than centralized nitrogen management via nitrification-denitrification if fertilizer production offsets are taken into account.

  7. CYLINDER OF THE DISPOSABLE MASS EXCHANGE DEVICE FOR HEMOSORPTION

    Directory of Open Access Journals (Sweden)

    F. I. Kazakov

    2015-01-01

    Full Text Available BACKGROUND. Hemocarboperfusion, previously widely used in our country, can universally pass out of use due to the lack of industrial production of disposable mass exchange devices.MATERIAl AND METHODS. Physicochemical properties of materials and design features of the body samples elements of various sizes have been studied.RESULTS. The elements and materials properties of the hemosorption mass exchanger cylinder have been studied. Hydrodynamic parameters of manipulation using the developed cylinders at different perfusion rates have been studied in bench experiments.CONCLUSION. The original cylinder of the disposable mass exchange device for hemosorption, which meets the current clinical needs, has been developed. 

  8. Hydraulic Validation of the LHC Cold Mass Heat Exchanger Tube

    CERN Document Server

    Provenaz, P

    1998-01-01

    The knowledge of the helium mass flow vs. the fraction of the tube wetted by the liquid helium II in the heat exchanger is a crucial input parameter for the heat exchange since the heat flux is direct ly proportional to the wetted surface. In the range of liquid and gas velocities inside the heat exchanger, the liquid flow behaves like in an open channel. Looking at the flow equations for such a s ituation, the velocity depends on the fluid properties only by the friction factor which is a function of the Reynolds number. Thus it was decided to build an experiment with water in order to check t he open channel equations in the heat exchanger geometry. This paper shows the results for water and gives the extrapolation for helium.

  9. Development of a unique decentralized rapid-response capability and contingency mass-casualty field hospital for the 1996 Democratic National Convention.

    Science.gov (United States)

    Binder, L S; Willoughby, P J; Matkaitis, L

    1997-01-01

    This article describes the planning, development, and execution of a unique, decentralized, and flexible medical response capability that was developed for the 1996 Democratic National Convention in Chicago. Concerns for coordinated acts of violence, terrorism, toxicologic exposures, and logistic problems posed by the United Center prompted the development of a decentralized and flexible rapid-response plan. Contingency planning for the remote possibility of a full-scale disaster led to the additional development of a contingency mass-casualty field hospital on site. The plans for this mass-gathering response are described in considerable detail. Forty-four patient encounters across the four days of the convention were recorded, with a combination of minor injuries and potentially serious medical presentations. The 1.46 EMS encounters per 1,000 attendees at the Democratic National Convention is comparable to other utilization rates for mass gatherings in the literature. Proactive attention to comprehensive contingency planning for equipment, supplies, personnel, and organizational needs, especially when multiagency response and cooperation are required, is essential.

  10. Radiocarbon detection by ion charge exchange mass spectrometry

    International Nuclear Information System (INIS)

    Hotchkis, Michael; Wei, Tao

    2007-01-01

    A method for detection of radiocarbon at low levels is described and the results of tests are presented. We refer to this method as ion charge exchange mass spectrometry (ICE-MS). The ICE-MS instrument is a two stage mass spectrometer. In the first stage, molecular interferences which would otherwise affect radiocarbon detection at mass 14 are eliminated by producing high charge state ions directly in the ion source (charge state ≥2). 14 N interference is eliminated in the second stage by converting the beam to negative ions in a charge exchange cell. The beam is mass-analysed at each stage. We have built a test apparatus consisting of an electron cyclotron resonance ion source and a pair of analysing magnets with a charge exchange cell in between, followed by an electrostatic analyser to improve the signal to background ratio. With this apparatus we have measured charge exchange probabilities for (C n+ → C - ) from 4.5 to 40.5 keV (n = 1-3). We have studied the sources of background including assessment of limits for nitrogen interference by searching for negative ions from charge exchange of 14 N ions. Our system has been used to detect 14 C in enriched samples of CO 2 gas with 14 C/ 12 C isotopic ratio down to the 10 -9 level. Combined with a measured sample consumption rate of 4 ng/s, this corresponds to a capability to detect transient signals containing only a few μBq of 14 C activity, such as may be obtained from chromatographic separation. The method will require further development to match the sensitivity of AMS with a gas ion source; however, even in its present state its sensitivity is well suited to tracer studies in biomedical research and drug development

  11. Considerations in the analysis of hydrogen exchange mass spectrometry data

    Science.gov (United States)

    Wales, Thomas E.; Eggertson, Michael J.; Engen, John R.

    2013-01-01

    i. Summary A major component of a hydrogen exchange mass spectrometry experiment is the analysis of protein and peptide mass spectra to yield information about deuterium incorporation. The processing of data that are produced includes the identification of each peptic peptide to create a master table/array of peptide sequence, retention time and retention time range, mass range and undeuterated mass. The amount of deuterium incorporated into each of the peptides in this array must then be determined. Various software platforms have been developed in order to perform this specific type of data analysis. We describe the fundamental parameters to be considered at each step along the way and how data processing, either by an individual or by software, must approach the analysis. PMID:23666730

  12. Minimizing Back Exchange in the Hydrogen Exchange-Mass Spectrometry Experiment

    Science.gov (United States)

    Walters, Benjamin T.; Ricciuti, Alec; Mayne, Leland; Englander, S. Walter

    2012-12-01

    The addition of mass spectrometry (MS) analysis to the hydrogen exchange (HX) proteolytic fragmentation experiment extends powerful HX methodology to the study of large biologically important proteins. A persistent problem is the degradation of HX information due to back exchange of deuterium label during the fragmentation-separation process needed to prepare samples for MS measurement. This paper reports a systematic analysis of the factors that influence back exchange (solution pH, ionic strength, desolvation temperature, LC column interaction, flow rates, system volume). The many peptides exhibit a range of back exchange due to intrinsic amino acid HX rate differences. Accordingly, large back exchange leads to large variability in D-recovery from one residue to another as well as one peptide to another that cannot be corrected for by reference to any single peptide-level measurement. The usual effort to limit back exchange by limiting LC time provides little gain. Shortening the LC elution gradient by 3-fold only reduced back exchange by ~2 %, while sacrificing S/N and peptide count. An unexpected dependence of back exchange on ionic strength as well as pH suggests a strategy in which solution conditions are changed during sample preparation. Higher salt should be used in the first stage of sample preparation (proteolysis and trapping) and lower salt (<20 mM) and pH in the second stage before electrospray injection. Adjustment of these and other factors together with recent advances in peptide fragment detection yields hundreds of peptide fragments with D-label recovery of 90 % ± 5 %.

  13. Minimizing back exchange in the hydrogen exchange-mass spectrometry experiment.

    Science.gov (United States)

    Walters, Benjamin T; Ricciuti, Alec; Mayne, Leland; Englander, S Walter

    2012-12-01

    The addition of mass spectrometry (MS) analysis to the hydrogen exchange (HX) proteolytic fragmentation experiment extends powerful HX methodology to the study of large biologically important proteins. A persistent problem is the degradation of HX information due to back exchange of deuterium label during the fragmentation-separation process needed to prepare samples for MS measurement. This paper reports a systematic analysis of the factors that influence back exchange (solution pH, ionic strength, desolvation temperature, LC column interaction, flow rates, system volume). The many peptides exhibit a range of back exchange due to intrinsic amino acid HX rate differences. Accordingly, large back exchange leads to large variability in D-recovery from one residue to another as well as one peptide to another that cannot be corrected for by reference to any single peptide-level measurement. The usual effort to limit back exchange by limiting LC time provides little gain. Shortening the LC elution gradient by 3-fold only reduced back exchange by ~2%, while sacrificing S/N and peptide count. An unexpected dependence of back exchange on ionic strength as well as pH suggests a strategy in which solution conditions are changed during sample preparation. Higher salt should be used in the first stage of sample preparation (proteolysis and trapping) and lower salt (<20 mM) and pH in the second stage before electrospray injection. Adjustment of these and other factors together with recent advances in peptide fragment detection yields hundreds of peptide fragments with D-label recovery of 90% ± 5%.

  14. Localized hydration in lyophilized myoglobin by hydrogen-deuterium exchange mass spectrometry. 2. Exchange kinetics.

    Science.gov (United States)

    Sophocleous, Andreas M; Topp, Elizabeth M

    2012-04-02

    Solid-state hydrogen-deuterium exchange with mass spectrometric analysis (ssHDX) is a promising method for characterizing proteins in amorphous solids. Though analysis of HDX kinetics is informative and well-established in solution, application of these methods to solid samples is complicated by possible heterogeneities in the solid. The studies reported here provide a detailed analysis of the kinetics of hydration and ssHDX for equine myoglobin (Mb) in solid matrices containing sucrose or mannitol. Water sorption was rapid relative to ssHDX, indicating that ssHDX kinetics was not limited by bulk water transport. Deuterium uptake in solids was well-characterized by a biexponential model; values for regression parameters provided insight into differences between the two solid matrices. Analysis of the widths of peptide mass envelopes revealed that, in solution, an apparent EX2 mechanism prevails, consistent with native conformation of the protein. In contrast, in mannitol-containing samples, a smaller non-native subpopulation exchanges by an EX1-like mechanism. Together, the results indicate that the analysis of ssHDX kinetic data and of the widths of peptide mass envelopes is useful in screening solid formulations of protein drugs for the presence of non-native species that cannot be detected by amide I FTIR.

  15. Corruption and Decentralization

    OpenAIRE

    Carbonara, Emanuela

    1999-01-01

    This paper studies the effect of decentralization on corruption in a hierarchical organization, where decentralization is intended as the delegation of control power to lower levels in a hierarchy. Decentral- ization causes a loss in control to the higher levels, thus curbing their incentives to monitor and detect corrupt activities. However, it also lowers the expected gains from corruption as, following decentralization, the number of individuals who are in charge of a single decision is re...

  16. On-tissue Direct Monitoring of Global Hydrogen/Deuterium Exchange by MALDI Mass Spectrometry: Tissue Deuterium Exchange Mass Spectrometry (TDXMS)*

    OpenAIRE

    Quanico, Jusal; Franck, Julien; Salzet, Michel; Fournier, Isabelle

    2016-01-01

    Hydrogen/deuterium exchange mass spectrometric (H/DXMS) methods for protein structural analysis are conventionally performed in solution. We present Tissue Deuterium Exchange Mass Spectrometry (TDXMS), a method to directly monitor deuterium uptake on tissue, as a means to better approximate the deuterium exchange behavior of proteins in their native microenvironment. Using this method, a difference in deuterium uptake behavior was observed when the same proteins were monitored in solution and...

  17. Localized hydration in lyophilized myoglobin by hydrogen-deuterium exchange mass spectrometry. 1. Exchange mapping.

    Science.gov (United States)

    Sophocleous, Andreas M; Zhang, Jun; Topp, Elizabeth M

    2012-04-02

    The local effects of hydration on myoglobin (Mb) in solid matrices containing mannitol or sucrose (1:1 w/w, protein:additive) were mapped using hydrogen-deuterium exchange with mass spectrometric analysis (HDX-MS) at 5 °C and compared to solution controls. Solid powders were exposed to D₂O(g) at controlled activity (a(w)) followed by reconstitution and analysis of the intact protein and peptides produced by pepsin digestion. HDX varied with matrix type, a(w), and position along the protein backbone. HDX was less in sucrose matrices than in mannitol matrices at all a(w) while the difference in solution was negligible. Differences in HDX in the two matrices were detectable despite similarities in their bulk water content. The extent of exchange in solids is proposed as a measure of the hydration of exchangeable amide groups, as well as protein conformation and dynamics; pepsin digestion allows these effects to be mapped with peptide-level resolution.

  18. The Area Between Exchange Curves as a Measure of Conformational Differences in Hydrogen-Deuterium Exchange Mass Spectrometry Studies

    Science.gov (United States)

    Mazur, Sharlyn J.; Weber, Daniel P.

    2017-05-01

    Hydrogen-deuterium exchange mass spectrometry (HDX-MS) provides information about protein conformational mobility under native conditions. The area between exchange curves, A bec , a functional data analysis concept, was adapted to the interpretation of HDX-MS data and provides a useful measure of exchange curve dissimilarity for tests of significance. Importantly, for most globular proteins under native conditions, A bec values provide an estimate of the log ratio of exchange-competent fractions in the two states, and thus are related to differences in the free energy of microdomain unfolding.

  19. Decentralized Software Architecture

    National Research Council Canada - National Science Library

    Khare, Rohit

    2002-01-01

    .... While the term "decentralization" is familiar from political and economic contexts, it has been applied extensively, if indiscriminately, to describe recent trends in software architecture towards...

  20. Recombinant Nepenthesin II for Hydrogen/Deuterium Exchange Mass Spectrometry.

    Science.gov (United States)

    Yang, Menglin; Hoeppner, Morgan; Rey, Martial; Kadek, Alan; Man, Petr; Schriemer, David C

    2015-07-07

    The pitcher secretions of the Nepenthes genus of carnivorous plants contain a proteolytic activity that is very useful for hydrogen/deuterium exchange mass spectrometry (HX-MS). Our efforts to reconstitute pitcher fluid activity using recombinant nepenthesin I (one of two known aspartic proteases in the fluid) revealed a partial cleavage profile and reduced enzymatic stability in certain HX-MS applications. We produced and characterized recombinant nepenthesin II to determine if it complemented nepenthesin I in HX-MS applications. Nepenthesin II shares many properties with nepenthesin I, such as fast digestion at reduced temperature and pH, and broad cleavage specificity, but in addition, it cleaves C-terminal to tryptophan. Neither enzyme reproduces the C-terminal proline cleavage we observed in the natural extract. Nepenthesin II is considerably more resistant to chemical denaturants and reducing agents than nepenthesin I, and it possesses a stability profile that is similar to that of pepsin. Higher stability combined with the slightly broader cleavage specificity makes nepenthesin II a useful alternative to pepsin and a more complete replacement for pitcher fluid in HX-MS applications.

  1. Decentralization: An International Perspective.

    Science.gov (United States)

    Walberg, Herbert J.; Paik, Susan J.; Komukai, Atsuko; Freeman, Karen

    2000-01-01

    Analysis of 22 definitions of decentralization shows how they are encompassed in the Organization for Economic Cooperation and Development's taxonomy. Data from 14 nations suggest that decentralization of instruction, planning, personnel management, and resource allocation and use has less effect on learning than do conditions in classrooms and…

  2. A Mass Computation Model for Lightweight Brayton Cycle Regenerator Heat Exchangers

    Science.gov (United States)

    Juhasz, Albert J.

    2010-01-01

    Based on a theoretical analysis of convective heat transfer across large internal surface areas, this paper discusses the design implications for generating lightweight gas-gas heat exchanger designs by packaging such areas into compact three-dimensional shapes. Allowances are made for hot and cold inlet and outlet headers for assembly of completed regenerator (or recuperator) heat exchanger units into closed cycle gas turbine flow ducting. Surface area and resulting volume and mass requirements are computed for a range of heat exchanger effectiveness values and internal heat transfer coefficients. Benefit cost curves show the effect of increasing heat exchanger effectiveness on Brayton cycle thermodynamic efficiency on the plus side, while also illustrating the cost in heat exchanger required surface area, volume, and mass requirements as effectiveness is increased. The equations derived for counterflow and crossflow configurations show that as effectiveness values approach unity, or 100 percent, the required surface area, and hence heat exchanger volume and mass tend toward infinity, since the implication is that heat is transferred at a zero temperature difference. To verify the dimensional accuracy of the regenerator mass computational procedure, calculation of a regenerator specific mass, that is, heat exchanger weight per unit working fluid mass flow, is performed in both English and SI units. Identical numerical values for the specific mass parameter, whether expressed in lb/(lb/sec) or kg/ (kg/sec), show the dimensional consistency of overall results.

  3. Subzero temperature chromatography for reduced back-exchange and improved dynamic range in amide hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Venable, John D; Okach, Linda; Agarwalla, Sanjay; Brock, Ansgar

    2012-11-06

    Amide hydrogen/deuterium exchange is a commonly used technique for studying the dynamics of proteins and their interactions with other proteins or ligands. When coupled with liquid chromatography and mass spectrometry, hydrogen/deuterium exchange provides several unique advantages over other structural characterization techniques including very high sensitivity, the ability to analyze proteins in complex environments, and a large mass range. A fundamental limitation of the technique arises from the loss of the deuterium label (back-exchange) during the course of the analysis. A method to limit loss of the label during the separation stage of the analysis using subzero temperature reversed-phase chromatography is presented. The approach is facilitated by the use of buffer modifiers that prevent freezing. We evaluated ethylene glycol, dimethyl formamide, formamide, and methanol for their freezing point suppression capabilities, effects on peptide retention, and their compatibilities with electrospray ionization. Ethylene glycol was used extensively because of its good electrospray ionization compatibility; however, formamide has potential to be a superior modifier if detrimental effects on ionization can be overcome. It is demonstrated using suitable buffer modifiers that separations can be performed at temperatures as low as -30 °C with negligible loss of the deuterium label, even during long chromatographic separations. The reduction in back-exchange is shown to increase the dynamic range of hydrogen/deuterium exchange mass spectrometry in terms of mixture complexity and the magnitude with which changes in deuteration level can be quantified.

  4. Advanced Heat/Mass Exchanger Technology for Geothermal and Solar Renewable Energy Systems

    Energy Technology Data Exchange (ETDEWEB)

    Greiner, Miles [Univ. of Nevada, Reno, NV (United States); Childress, Amy [Univ. of Nevada, Reno, NV (United States); Hiibel, Sage [Univ. of Nevada, Reno, NV (United States); Kim, Kwang [Univ. of Nevada, Reno, NV (United States); Park, Chanwoo [Univ. of Nevada, Reno, NV (United States); Wirtz, Richard [Univ. of Nevada, Reno, NV (United States)

    2014-12-16

    Northern Nevada has abundant geothermal and solar energy resources, and these renewable energy sources provide an ample opportunity to produce economically viable power. Heat/mass exchangers are essential components to any energy conversion system. Improvements in the heat/mass exchange process will lead to smaller, less costly (more efficient) systems. There is an emerging heat transfer technology, based on micro/nano/molecular-scale surface science that can be applied to heat/mass exchanger design. The objective is to develop and characterize unique coating materials, surface configurations and membranes capable of accommodating a 10-fold increase in heat/mass exchanger performance via phase change processes (boiling, condensation, etc.) and single phase convective heat/mass transfer.

  5. Decentralized Online Social Networks

    Science.gov (United States)

    Datta, Anwitaman; Buchegger, Sonja; Vu, Le-Hung; Strufe, Thorsten; Rzadca, Krzysztof

    Current Online social networks (OSN) are web services run on logically centralized infrastructure. Large OSN sites use content distribution networks and thus distribute some of the load by caching for performance reasons, nevertheless there is a central repository for user and application data. This centralized nature of OSNs has several drawbacks including scalability, privacy, dependence on a provider, need for being online for every transaction, and a lack of locality. There have thus been several efforts toward decentralizing OSNs while retaining the functionalities offered by centralized OSNs. A decentralized online social network (DOSN) is a distributed system for social networking with no or limited dependency on any dedicated central infrastructure. In this chapter we explore the various motivations of a decentralized approach to online social networking, discuss several concrete proposals and types of DOSN as well as challenges and opportunities associated with decentralization.

  6. Decentralized energy studies: bibliography

    Energy Technology Data Exchange (ETDEWEB)

    Ohi, J.M.; Unseld, C.T.; Levine, A.; Silversmith, J.A.

    1980-05-01

    This bibliography is a compilation of literature on decentralized energy systems. It is arranged according to topical (e.g., lifestyle and values, institutions, and economics) and geographical scale to facilitate quick reference to specific areas of interest. Also included are articles by and about Amory B. Lovins who has played a pivotal role in making energy decentralization an important topic of national debate. Periodicals, other bibliographies, and directories are also listed.

  7. On-tissue Direct Monitoring of Global Hydrogen/Deuterium Exchange by MALDI Mass Spectrometry: Tissue Deuterium Exchange Mass Spectrometry (TDXMS)*

    Science.gov (United States)

    Quanico, Jusal; Franck, Julien

    2016-01-01

    Hydrogen/deuterium exchange mass spectrometric (H/DXMS) methods for protein structural analysis are conventionally performed in solution. We present Tissue Deuterium Exchange Mass Spectrometry (TDXMS), a method to directly monitor deuterium uptake on tissue, as a means to better approximate the deuterium exchange behavior of proteins in their native microenvironment. Using this method, a difference in deuterium uptake behavior was observed when the same proteins were monitored in solution and on tissue. The higher maximum deuterium uptake at equilibrium for all proteins analyzed in solution suggests a more open conformation in the absence of interacting partners normally observed on tissue. We also demonstrate a difference in the deuterium uptake behavior of a few proteins across different morphological regions of the same tissue section. Modifications of the total number of hydrogens exchanged, as well as the kinetics of exchange, were both observed. These results provide information on the implication of protein interactions with partners as well as on the conformational changes related to these interactions, and illustrate the importance of examining protein deuterium exchange behavior in the presence of its specific microenvironment directly at the level of tissues. PMID:27512083

  8. On-tissue Direct Monitoring of Global Hydrogen/Deuterium Exchange by MALDI Mass Spectrometry: Tissue Deuterium Exchange Mass Spectrometry (TDXMS).

    Science.gov (United States)

    Quanico, Jusal; Franck, Julien; Salzet, Michel; Fournier, Isabelle

    2016-10-01

    Hydrogen/deuterium exchange mass spectrometric (H/DXMS) methods for protein structural analysis are conventionally performed in solution. We present Tissue Deuterium Exchange Mass Spectrometry (TDXMS), a method to directly monitor deuterium uptake on tissue, as a means to better approximate the deuterium exchange behavior of proteins in their native microenvironment. Using this method, a difference in deuterium uptake behavior was observed when the same proteins were monitored in solution and on tissue. The higher maximum deuterium uptake at equilibrium for all proteins analyzed in solution suggests a more open conformation in the absence of interacting partners normally observed on tissue. We also demonstrate a difference in the deuterium uptake behavior of a few proteins across different morphological regions of the same tissue section. Modifications of the total number of hydrogens exchanged, as well as the kinetics of exchange, were both observed. These results provide information on the implication of protein interactions with partners as well as on the conformational changes related to these interactions, and illustrate the importance of examining protein deuterium exchange behavior in the presence of its specific microenvironment directly at the level of tissues. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. Online hydrogen/deuterium exchange performed in the ion mobility cell of a hybrid mass spectrometer.

    Science.gov (United States)

    Nagy, Kornél; Redeuil, Karine; Rezzi, Serge

    2009-11-15

    The present paper describes the performance of online, gas-phase hydrogen/deuterium exchange implemented in the ion mobility cell of a quadrupole time-of-flight mass spectrometer. Deuterium oxide and deuterated methanol were utilized to create deuterated vapor that is introduced into the ion mobility region of the mass spectrometer. Hydrogen/deuterium exchange occurs spontaneously in the milliseconds time frame without the need of switching the instrument into ion mobility mode. The exchange was studied in case of low molecular weight molecules and proteins. The observed number of exchanged hydrogens was equal to the number of theoretically exchangeable hydrogens for all low molecular weight compounds. This method needs only minimal instrumental modifications, is simple, cheap, environment friendly, compatible with ultraperformance liquid chromatography, and can be implemented on commercially available instruments. It does not compromise choice of liquid chromatographic solvents and accurate mass or parallel-fragmentation (MS(E)) methods. The performance of this method was compared to that of conventional alternatives where the deuterated solvent is introduced into the cone gas of the instrument. Although the degree of exchange was similar between the two methods, the "cone gas method" requires 10 times higher deuterated solvent volumes (50 muL/min) and offers reduced sensitivity in the tandem mass spectrometry (MS/MS) mode. The presented method is suggested as a standard future element of mass spectrometers to aid online structural characterization of unknowns and to study conformational changes of proteins with hydrogen/deuterium exchange.

  10. Hydrogen/deuterium exchange in electrospray ionization mass spectrometry as a practical tool for structure elucidation

    International Nuclear Information System (INIS)

    Conboy, J.J.; Bean, M.F.; Hemling, M.E.; Carr, S.A.

    1993-01-01

    Measurement of the number of active hydrogens in a molecule by various types of hydrogen/deuterium exchange experiments has for a number of years proven useful in the elucidation of compounds of unknown structure, in the determination of the mechanism of chemical or biological reactions, and in the interpretation of fragmentation pathways in mass spectra. Recent work in the analysis of protein conformation by ESIMS has made use of both solution phase and gas phase (atmospheric pressure or in vacuo) hydrogen/deuterium exchange methods. The gas phase exchange experiments were performed either on FTMS instrumentation to achieve the long residence times required for exchange or with a heated dual-capillary inlet reactor interface for desolvation and then exchange of the sample molecules. The authors have implemented methods for hydrogen/deuterium exchange which require only simple changes to the plumbing of a Sciex API-III Ionspray source. Effective exchange of active hydrogens may be achieved by replacing either all of the nebulizer gas or part of the curtain gas or both with a deuterated exchange agent such as ND 3 . These methods provide for essentially complete exchange of all active hydrogens on the types of small molecules (e.g., alkaloids, glycosides, etc.) for which H/D exchange has been of proven utility in structure elucidation work; larger molecules, or at least those with a large number of potentially exchangeable hydrogens, proved to be more challenging particularly in regard to completeness of exchange. These methods also appear to work without the need for dissolving the sample in deuterated or aprotic solvents. A simple ESIMS H/D exchange experiment is a useful addition to the mass spectrometrist's armamentarium for the elucidation of unknown structures. With it, H/D exchange is now extended to classes of compounds not amenable to FAB or DCI, especially in cases where sample is limited

  11. SAIDE: A Semi-Automated Interface for Hydrogen/Deuterium Exchange Mass Spectrometry

    OpenAIRE

    Villar, Maria T.; Miller, Danny E.; Fenton, Aron W.; Artigues, Antonio

    2010-01-01

    Deuterium/hydrogen exchange in combination with mass spectrometry (DH MS) is a sensitive technique for detection of changes in protein conformation and dynamics. Since temperature, pH and timing control are the key elements for reliable and efficient measurement of hydrogen/deuterium content in proteins and peptides, we have developed a small, semiautomatic interface for deuterium exchange that interfaces the HPLC pumps with a mass spectrometer. This interface is relatively inexpensive to bui...

  12. Vertical air mass exchange driven by the local circulation on the northern slope of Mount Everest

    Science.gov (United States)

    Zhou, Libo; Zou, Han; Ma, Shupo; Li, Peng; Zhu, Jinhuan; Huo, Cuiping

    2011-01-01

    To better understand vertical air mass exchange driven by local circulation in the Himalayas, the volume flux of air mass is estimated in the Rongbuk Valley on the northern slope of Mount Everest, based on a volume closure method and wind-profiler measurements during the HEST2006 campaign in June 2006. Vertical air mass exchange was found to be dominated by a strong downward mass transfer from the late morning to late night. The average vertical air volume flux was 0.09 m s-1, which could be equivalent to a daily ventilation of 30 times the enclosed valley volume. This vertical air mass exchange process was greatly affected by the evolution of the South Asian summer monsoon (SASM), with a strong downward transfer during the SASM break stage, and a weak transfer during the SASM active stage.

  13. Transport and mass exchange processes in sand and gravel aquifers (v.1)

    International Nuclear Information System (INIS)

    Moltyaner, G.

    1990-01-01

    The objectives of this conference were to exchange information on promising field measurement techniques used for the characterization of spatial variability of geologic formations and on new methods used for quantifying the effect of spatial variability on groundwater flow and transport of materials; to discuss novel developments in the theory of transport processes and simulation methods; and to present views and opinions on future initiatives and directions in the design of large-scale field tracer experiments and the development of conceptual and mathematical models of transport and mass exchange processes. The 46 papers presented in these proceedings are divided into six sections: field studies of transport processes; groundwater tracers and novel field measurement techniques; promising methods and field measurement techniques for quantifying the effect of geological heterogeneities on groundwater flow and transport; novel developments in the theory of transport processes; numerical modelling of transport and mass exchange processes; and field and modelling studies of mass exchange processes. (L.L.)

  14. Decentralized Ground Staff Scheduling

    DEFF Research Database (Denmark)

    Sørensen, M. D.; Clausen, Jens

    2002-01-01

    Typically, ground staff scheduling is centrally planned for each terminal in an airport. The advantage of this is that the staff is efficiently utilized, but a disadvantage is that staff spends considerable time walking between stands. In this paper a decentralized approach for ground staff...... scheduling is investigated. The airport terminal is divided into zones, where each zone consists of a set of stands geographically next to each other. Staff is assigned to work in only one zone and the staff scheduling is planned decentralized for each zone. The advantage of this approach is that the staff...... work in a smaller area of the terminal and thus spends less time walking between stands. When planning decentralized the allocation of stands to flights influences the staff scheduling since the workload in a zone depends on which flights are allocated to stands in the zone. Hence solving the problem...

  15. Decentralization and Governance in Indonesia

    NARCIS (Netherlands)

    Holzhacker, Ronald; Wittek, Rafael; Woltjer, Johan

    2016-01-01

    I. Theoretical Reflections on Decentralization and Governance for Sustainable Society 1. Decentralization and Governance for Sustainable Society in Indonesia Ronald Holzhacker, Rafael Wittek and Johan Woltjer 2. Good Governance Contested: Exploring Human Rights and Sustainability as Normative Goals

  16. Bitcoin as a decentralized currency

    Directory of Open Access Journals (Sweden)

    Dinić Vladimir

    2014-01-01

    Full Text Available Bitcoin is the first decentralized peer-to-peer crypto-currency founded in 2009. Its main specificity is the fact that there is no issuer of this currency. On the other hand, the supply of this currency is software-programmed and limited. Among other things, its main features are relatively secure payments, low transaction costs, anonymity, inability of counterfeiting, irreversibility of transactions, but also extremely unstable exchange rate. Despite many advantages, the use of this currency is subject of numerous discussions, as this currency offers the possibility of performing various abuses and criminal activities. The future of this and other currencies in this regard depends on both security and privacy of these currencies, and legal regulation of such payments.

  17. Measuring the hydrogen/deuterium exchange of proteins at high spatial resolution by mass spectrometry

    DEFF Research Database (Denmark)

    Rand, Kasper Dyrberg; Zehl, Martin; Jørgensen, Thomas J D

    2014-01-01

    the sum of the exchange kinetics for the individual backbone amides. Local exchange kinetics is typically achieved by using pepsin digestion under quench conditions (i.e., under cold acidic conditions where the amide hydrogen exchange rate is slowed by many orders of magnitude). The ability to localize...... resolution of the lowest-energy structure of the native ensemble. There is a growing need for sensitive analytical tools to explore all of the significant molecular structures in the conformational landscape of proteins. Hydrogen/deuterium exchange monitored by mass spectrometry (HDX-MS) has recently emerged...... as a powerful method for characterizing protein conformational dynamics. The basis of this method is the fact that backbone amides in stable hydrogen-bonded structures (e.g., α-helices and β-sheets) are protected against exchange with the aqueous solvent. All protein structures are dynamic, however...

  18. Decentralized control: An overview

    Czech Academy of Sciences Publication Activity Database

    Bakule, Lubomír

    2008-01-01

    Roč. 32, č. 1 (2008), s. 87-98 ISSN 1367-5788 R&D Projects: GA AV ČR(CZ) IAA200750802; GA MŠk(CZ) LA 282 Institutional research plan: CEZ:AV0Z10750506 Keywords : decentralized control * large-scale systems * decomposition Subject RIV: BC - Control Systems Theory Impact factor: 1.109, year: 2008

  19. Decentralized control and communication

    Czech Academy of Sciences Publication Activity Database

    Bakule, Lubomír; Papík, Martin

    2012-01-01

    Roč. 36, č. 1 (2012), s. 1-10 ISSN 1367-5788 R&D Projects: GA MŠk(CZ) LG12014 Institutional research plan: CEZ:AV0Z10750506 Keywords : decentralization * communication * large-scale complex systems Subject RIV: BC - Control Systems Theory Impact factor: 1.289, year: 2012

  20. Coalition or decentralization

    DEFF Research Database (Denmark)

    Mahdiraji, Hannan Amoozad; Govindan, Kannan; Zavadskas, Edmundas Kazimieras

    2014-01-01

    retailers. The Nash equilibrium and definition are used bearing in mind inventory and pricing and marketing cost as decision variables for this matter. This paper studies a three-echelon supply chain network and focuses on the value of integrating a pair of partners in the chain. In the decentralized case...

  1. Intracellular Water Exchange for Measuring the Dry Mass, Water Mass and Changes in Chemical Composition of Living Cells

    Science.gov (United States)

    Hecht, Vivian C.; Son, Sungmin; Li, Yingzhong; Knudsen, Scott M.; Olcum, Selim; Higgins, John M.; Chen, Jianzhu; Grover, William H.; Manalis, Scott R.

    2013-01-01

    We present a method for direct non-optical quantification of dry mass, dry density and water mass of single living cells in suspension. Dry mass and dry density are obtained simultaneously by measuring a cell’s buoyant mass sequentially in an H2O-based fluid and a D2O-based fluid. Rapid exchange of intracellular H2O for D2O renders the cell’s water content neutrally buoyant in both measurements, and thus the paired measurements yield the mass and density of the cell’s dry material alone. Utilizing this same property of rapid water exchange, we also demonstrate the quantification of intracellular water mass. In a population of E. coli, we paired these measurements to estimate the percent dry weight by mass and volume. We then focused on cellular dry density – the average density of all cellular biomolecules, weighted by their relative abundances. Given that densities vary across biomolecule types (RNA, DNA, protein), we investigated whether we could detect changes in biomolecular composition in bacteria, fungi, and mammalian cells. In E. coli, and S. cerevisiae, dry density increases from stationary to exponential phase, consistent with previously known increases in the RNA/protein ratio from up-regulated ribosome production. For mammalian cells, changes in growth conditions cause substantial shifts in dry density, suggesting concurrent changes in the protein, nucleic acid and lipid content of the cell. PMID:23844039

  2. Intracellular water exchange for measuring the dry mass, water mass and changes in chemical composition of living cells.

    Science.gov (United States)

    Feijó Delgado, Francisco; Cermak, Nathan; Hecht, Vivian C; Son, Sungmin; Li, Yingzhong; Knudsen, Scott M; Olcum, Selim; Higgins, John M; Chen, Jianzhu; Grover, William H; Manalis, Scott R

    2013-01-01

    We present a method for direct non-optical quantification of dry mass, dry density and water mass of single living cells in suspension. Dry mass and dry density are obtained simultaneously by measuring a cell's buoyant mass sequentially in an H2O-based fluid and a D2O-based fluid. Rapid exchange of intracellular H2O for D2O renders the cell's water content neutrally buoyant in both measurements, and thus the paired measurements yield the mass and density of the cell's dry material alone. Utilizing this same property of rapid water exchange, we also demonstrate the quantification of intracellular water mass. In a population of E. coli, we paired these measurements to estimate the percent dry weight by mass and volume. We then focused on cellular dry density - the average density of all cellular biomolecules, weighted by their relative abundances. Given that densities vary across biomolecule types (RNA, DNA, protein), we investigated whether we could detect changes in biomolecular composition in bacteria, fungi, and mammalian cells. In E. coli, and S. cerevisiae, dry density increases from stationary to exponential phase, consistent with previously known increases in the RNA/protein ratio from up-regulated ribosome production. For mammalian cells, changes in growth conditions cause substantial shifts in dry density, suggesting concurrent changes in the protein, nucleic acid and lipid content of the cell.

  3. MACROECONOMIC IMPACT OF DECENTRALIZATION

    Directory of Open Access Journals (Sweden)

    Emilia Cornelia STOICA

    2014-05-01

    Full Text Available The concept of decentralization has a variety of expressions, but the meaning generally accepted refers to the transfer of authority and responsibility of the public functions from central government to sub-national public entities or even to the private sector. Decentralization process is complex, affecting many aspects of social and economic life and public management, and its design and implementation cover several stages, depending on the cyclical and structural developments of the country. From an economic perspective, decentralization is seen as a means of primary importance in terms of improving the effectiveness and efficiency of public services and macroeconomic stability due to the redistribution of public finances while in a much closer logic of the government policy objectives. But the decentralization process behaves as well some risks, because it involves the implementation of appropriate mechanisms for the establishment of income and expenditure programming at the subnational level, which, if is not correlated with macroeconomic policy imperatives can lead to major imbalances, both financially as in termes of economic and social life. Equally, ensuring the balance of the budget at the local level is imperative to fulfill, this goal imposing a legal framework and specific procedures to size transfers of public funds, targeted or untargeted. Also, public and local authorities have to adopt appropriate laws and regulations such that sub-national public entities can access loans - such as bank loans or debentures from domestic or external market - in terms of a strict monitoring national financial stability. In all aspects of decentralization - political, administrative, financial -, public authorities should develop and implement the most effective mechanisms to coordinate macroeconomic objectives and both sectoral and local interests and establish clear responsibilities - exclusive or shared - for all parties involved in the

  4. Intensification of heat and mass transfer by ultrasound: application to heat exchangers and membrane separation processes.

    Science.gov (United States)

    Gondrexon, N; Cheze, L; Jin, Y; Legay, M; Tissot, Q; Hengl, N; Baup, S; Boldo, P; Pignon, F; Talansier, E

    2015-07-01

    This paper aims to illustrate the interest of ultrasound technology as an efficient technique for both heat and mass transfer intensification. It is demonstrated that the use of ultrasound results in an increase of heat exchanger performances and in a possible fouling monitoring in heat exchangers. Mass transfer intensification was observed in the case of cross-flow ultrafiltration. It is shown that the enhancement of the membrane separation process strongly depends on the physico-chemical properties of the filtered suspensions. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Electrochemical reduction of disulfide-containing proteins for hydrogen/deuterium exchange monitored by mass spectrometry

    DEFF Research Database (Denmark)

    Mysling, Simon; Salbo, Rune; Ploug, Michael

    2014-01-01

    requires a high concentration (>200 mM) of the chemical reducing agent Tris(2-carboxyethyl)phosphine (TCEP) as the reduction rate constant is decreased at low pH and temperature. Serious adverse effects on chromatographic and mass spectrometric performances have been reported when using high concentrations......Characterization of disulfide bond-containing proteins by hydrogen/deuterium exchange monitored by mass spectrometry (HDX-MS) requires reduction of the disulfide bonds under acidic and cold conditions, where the amide hydrogen exchange reaction is quenched (pH 2.5, 0°C). The reduction typically...

  6. Mapping Protein-Ligand Interactions with Proteolytic Fragmentation, Hydrogen/Deuterium Exchange-Mass Spectrometry.

    Science.gov (United States)

    Gallagher, Elyssia S; Hudgens, Jeffrey W

    2016-01-01

    Biological processes are the result of noncovalent, protein-ligand interactions, where the ligands range from small organic and inorganic molecules to lipids, nucleic acids, peptides, and proteins. Amide groups within proteins constantly exchange protons with water. When immersed in heavy water (D2O), mass spectrometry (MS) can measure the change of mass associated with the hydrogen to deuterium exchange (HDX). Protein-ligand interactions modify the hydrogen exchange rates of amide protons, and the measurement of the amide exchange rates can provide rich information regarding the dynamical structure of the protein-ligand complex. This chapter describes a protocol for conducting bottom-up, continuous uptake, proteolytic fragmentation HDX-MS experiments that can help identify and map the interacting peptides of a protein-ligand interface. This tutorial outlines the fundamental theory governing hydrogen exchange; provides practical information regarding the preparation of protein samples and solutions; and describes the exchange reaction, reaction quenching, enzymatic digestion, chromatographic separation, and peptide analysis by MS. Tables list representative combinations of fluidic components used by HDX-MS researchers and summarize the available HDX-MS analysis software packages. Additionally, two HDX-MS case studies are used to illustrate protein-ligand interactions involving: (1) a continuous sequence of interacting residues and (2) a set of discontinuously numbered residues, residing spatially near each other. © 2016 Elsevier Inc. All rights reserved.

  7. Probing protein ligand interactions by automated hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Chalmers, Michael J; Busby, Scott A; Pascal, Bruce D; He, Yuanjun; Hendrickson, Christopher L; Marshall, Alan G; Griffin, Patrick R

    2006-02-15

    Amide hydrogen/deuterium exchange is a powerful biophysical technique for probing changes in protein dynamics induced by ligand interaction. The inherent low throughput of the technology has limited its impact on drug screening and lead optimization. Automation increases the throughput of H/D exchange to make it compatible with drug discovery efforts. Here we describe the first fully automated H/D exchange system that provides highly reproducible H/D exchange kinetics from 130 ms to 24 h. Throughput is maximized by parallel sample processing, and the system can run H/D exchange assays in triplicate without user intervention. We demonstrate the utility of this system to differentiate structural perturbations in the ligand-binding domain (LBD) of the nuclear receptor PPARgamma induced upon binding a full agonist and a partial agonist. PPARgamma is the target of glitazones, drugs used for treatment of insulin resistance associated with type II diabetes. Recently it has been shown that partial agonists of PPARgamma have insulin sensitization properties while lacking several adverse effects associated with full agonist drugs. To further examine the mechanism of partial agonist activation of PPARgamma, we extended our studies to the analysis of ligand interactions with the heterodimeric complex of PPARgamma/RXRalpha LBDs. To facilitate analysis of H/D exchange of large protein complexes, we performed the experiment with a 14.5-T Fourier transform ion cyclotron resonance mass spectrometer capable of measuring mass with accuracy in the ppb range.

  8. Asynchronous decentralized method for interconnected electricity markets

    International Nuclear Information System (INIS)

    Huang, Anni; Joo, Sung-Kwan; Song, Kyung-Bin; Kim, Jin-Ho; Lee, Kisung

    2008-01-01

    This paper presents an asynchronous decentralized method to solve the optimization problem of interconnected electricity markets. The proposed method decomposes the optimization problem of combined electricity markets into individual optimization problems. The impact of neighboring markets' information is included in the objective function of the individual market optimization problem by the standard Lagrangian relaxation method. Most decentralized optimization methods use synchronous models of communication to exchange updated market information among markets during the iterative process. In this paper, however, the solutions of the individual optimization problems are coordinated through an asynchronous communication model until they converge to the global optimal solution of combined markets. Numerical examples are presented to demonstrate the advantages of the proposed asynchronous method over the existing synchronous methods. (author)

  9. South Atlantic interbasin exchanges of mass, heat, salt and anthropogenic carbon

    NARCIS (Netherlands)

    Evans, G.R.; McDonagh, E.L.; King, B.A.; Bryden, H.L.; Bakker, D.C.E.; Brown, P.J.; Schuster, U.; Speer, K.G.; van Heuven, S.M.A.C.

    2017-01-01

    The exchange of mass, heat, salt and anthropogenic carbon (Cant) between the South Atlantic, south of 24°S, and adjacent ocean basins is estimated from hydrographic data obtained during 2008–2009 using an inverse method. Transports of anthropogenic carbon are calculated across the western (Drake

  10. Mixture theory for a thermoelasto-plastic porous solid considering fluid flow and internal mass exchange

    DEFF Research Database (Denmark)

    Ristinmaa, M.; Ottosen, N.S.; Johannesson, Björn

    2011-01-01

    A thermoelastic-plastic body consisting of two phases, a solid and a fluid, each comprising two constituents is considered where one constituent in one phase is allowed to exchange mass with another constituent (of the same substance) in the other phase. A large strain setting is adopted...

  11. Decentralized Portfolio Management

    Directory of Open Access Journals (Sweden)

    Benjamin Miranda Tabak

    2003-12-01

    Full Text Available We use a mean-variance model to analyze the problem of decentralized portfolio management. We find the solution for the optimal portfolio allocation for a head trader operating in n different markets, which is called the optimal centralized portfolio. However, as there are many traders specialized in different markets, the solution to the problem of optimal decentralized allocation should be different from the centralized case. In this paper we derive conditions for the solutions to be equivalent. We use multivariate normal returns and a negative exponential function to solve the problem analytically. We generate the equivalence of solutions by assuming that different traders face different interest rates for borrowing and lending. This interest rate is dependent on the ratio of the degrees of risk aversion of the trader and the head trader, on the excess return, and on the correlation between asset returns.

  12. Policy Implementation Decentralization Government in Indonesia

    Directory of Open Access Journals (Sweden)

    Kardin M. Simanjuntak

    2015-06-01

    Full Text Available Decentralization in Indonesia is that reforms not completed and until the current implementation is not maximized or have not been successful. The essence of decentralization is internalising cost and benefit' for the people and how the government closer to the people. That's the most important essence of essence 'decentralization’. However, the implementation of decentralization in Indonesia is still far from the expectations. It is shown that only benefits of decentralization elite and local authorities, decentralization is a neo-liberal octopus, decentralization of public services are lacking in character, decentralization without institutional efficiency, decentralization fosters corruption in the area, and quasi-fiscal decentralization.

  13. The mass balance of a Proton Exchange Membrane Fuel Cell (PEMFC)

    International Nuclear Information System (INIS)

    Miloud, S.; Kamaruzzaman Sopian; Wan Ramli Wan Daud

    2006-01-01

    A Proton Exchange Membrane Fuel Cell (PEMFC), operating at low temperature uses a simple chemical process to combine hydrogen and oxygen into water, producing electric current and heat during the electrochemical reaction. This work concern on the theoretical consideration of the mass balance has been evaluated to predict the mass flow rate of the both gases (hydrogen/oxygen), the water mass balance, and the heat transfer in order to design a single cell PEMFC stack with a better flow field distributor on the performance of Polymer Electrolyte membrane fuel cells

  14. Decentralized control of complex systems

    CERN Document Server

    Siljak, Dragoslav D

    2011-01-01

    Complex systems require fast control action in response to local input, and perturbations dictate the use of decentralized information and control structures. This much-cited reference book explores the approaches to synthesizing control laws under decentralized information structure constraints.Starting with a graph-theoretic framework for structural modeling of complex systems, the text presents results related to robust stabilization via decentralized state feedback. Subsequent chapters explore optimization, output feedback, the manipulative power of graphs, overlapping decompositions and t

  15. Sub-Zero Temperature Chromatography for Reduced Back-Exchange and Improved Dynamic Range in Amide Hydrogen Deuterium Exchange Mass Spectrometry

    OpenAIRE

    Venable, John D.; Okach, Linda; Agarwalla, Sanjay; Brock, Ansgar

    2012-01-01

    Amide hydrogen/deuterium exchange is a commonly used technique for studying the dynamics of proteins and their interactions with other proteins or ligands. When coupled with liquid chromatography and mass spectrometry, hydrogen/deuterium exchange provides several unique advantages over other structural characterization techniques including very high sensitivity, the ability to analyze proteins in complex environments, and a large mass range. A fundamental limitation of the technique arises fr...

  16. Ion mobility spectrometry-hydrogen deuterium exchange mass spectrometry of anions: part 1. Peptides to proteins.

    Science.gov (United States)

    Donohoe, Gregory C; Khakinejad, Mahdiar; Valentine, Stephen J

    2015-04-01

    Ion mobility spectrometry (IMS) coupled with hydrogen deuterium exchange (HDX)-mass spectrometry (MS) has been used to study the conformations of negatively-charged peptide and protein ions. Results are presented for ion conformers of angiotensin 1, a synthetic peptide (SP), bovine insulin, ubiquitin, and equine cytochrome c. In general, the SP ion conformers demonstrate a greater level of HDX efficiency as a greater proportion of the sites undergo HDX. Additionally, these ions exhibit the fastest rates of exchange. Comparatively, the angiotensin 1 ions exhibit a lower rate of exchange and HDX level presumably because of decreased accessibility of exchange sites by charge sites. The latter are likely confined to the peptide termini. Insulin ions show dramatically reduced HDX levels and exchange rates, which can be attributed to decreased conformational flexibility resulting from the disulfide bonds. For the larger ubiquitin and protein ions, increased HDX is observed for larger ions of higher charge state. For ubiquitin, a conformational transition from compact to more elongated species (from lower to higher charge states) is reflected by an increase in HDX levels. These results can be explained by a combination of interior site protection by compact conformers as well as decreased access by charge sites. The elongated cytochrome c ions provide the largest HDX levels where higher values correlate with charge state. These results are consistent with increased exchange site accessibility by additional charge sites. The data from these enhanced IMS-HDX experiments are described in terms of charge site location, conformer rigidity, and interior site protection.

  17. A mass spectrometric investigation of the exchange of deuterium with tetraborane(10) in the gas phase

    International Nuclear Information System (INIS)

    Greatrex, R.; Greenwood, N.N.; Potter, C.D.

    1984-01-01

    Mass spectrometry has been used to establish direct exchange between dideuterium (D 2 ) and B 4 H 10 in the gas phase. In a mixture of stoicheiometry B 4 H 10 :3D 2 at 42 deg C, ca. 7% of the hydrogen atoms were replaced by deuterium in 130 min. The possibility that the exchange might be occurring by an indirect or autocatalytic mechanism was eliminated by a series of experiments in which the rate of exchange was shown to be independent of small admixtures of the decomposition products. The present work resolves a troublesome inconsistency in the literature and strengthens the view that the equilibrium B 4 H 10 brace B 4 H 8 brace + H 2 plays a key role in borane interconversion reactions. (author)

  18. Enhancement of combined heat and mass transfer in a vertical-tube heat and mass exchanger

    International Nuclear Information System (INIS)

    Webb, R.L.; Perez-Blanco, H.

    1986-01-01

    This paper studies enhancement of heat and mass transfer between a countercurrent, gravity-drained water film and air flowing in a vertical tube. The enhancement technique employed is spaced, transverse wires placed in the air boundary layer, near the air--water interface. Heat transfer correlations for turbulent, single-phase heat transfer in pipes having wall-attached spaced ribs are used to select the preferred wire diameter, and to predict the gas phase heat and mass transfer coefficients. Tests were run with two different radial placements of the rib roughness: (1) at the free surface of the liquid film, and (2) the base of the roughness displaced 0.51 mm into the air flow. The authors hypothesize that the best heat/mass transfer and friction performance will be obtained with the roughness at the surface of the water film. Experiments conducted with both roughness placements show that the authors' hypothesis is correct. The measured heat/mass transfer enhancement agreed very closely with the predicted values. A unique feature of the enhancement concept is that it does not require surface wetting of the enhancement device to provide enhancement

  19. Hydrogen/deuterium exchange mass spectrometry applied to IL-23 interaction characteristics: potential impact for therapeutics.

    Science.gov (United States)

    Iacob, Roxana E; Krystek, Stanley R; Huang, Richard Y-C; Wei, Hui; Tao, Li; Lin, Zheng; Morin, Paul E; Doyle, Michael L; Tymiak, Adrienne A; Engen, John R; Chen, Guodong

    2015-04-01

    IL-23 is an important therapeutic target for the treatment of inflammatory diseases. Adnectins are targeted protein therapeutics that are derived from domain III of human fibronectin and have a similar protein scaffold to antibodies. Adnectin 2 was found to bind to IL-23 and compete with the IL-23/IL-23R interaction, posing a potential protein therapeutic. Hydrogen/deuterium exchange mass spectrometry and computational methods were applied to probe the binding interactions between IL-23 and Adnectin 2 and to determine the correlation between the two orthogonal methods. This review summarizes the current structural knowledge about IL-23 and focuses on the applicability of hydrogen/deuterium exchange mass spectrometry to investigate the higher order structure of proteins, which plays an important role in the discovery of new and improved biotherapeutics.

  20. HDXFinder: Automated Analysis and Data Reporting of Deuterium/Hydrogen Exchange Mass Spectrometry

    Science.gov (United States)

    Miller, Danny E.; Prasannan, Charulata B.; Villar, Maria T.; Fenton, Aron W.; Artigues, Antonio

    2012-02-01

    Hydrogen/deuterium exchange in combination with mass spectrometry (H/D MS) is a sensitive technique for detection of changes in protein conformation and dynamics. However, wide application of H/D MS has been hindered, in part, by the lack of computational tools necessary for efficient analysis of the large data sets associated with this technique. We report a novel web-based application for automatic analysis of H/D MS experimental data. This application relies on the high resolution of mass spectrometers to extract all isotopic envelopes before correlating these envelopes with individual peptides. Although a fully automatic analysis is possible, a variety of graphical tools are included to aid in the verification of correlations and rankings of the isotopic peptide envelopes. As a demonstration, the rate constants for H/D exchange of peptides from rabbit muscle pyruvate kinase are mapped onto the structure of this protein.

  1. Actin Isoform-specific Conformational Differences Observed with Hydrogen/Deuterium Exchange and Mass Spectrometry*

    Science.gov (United States)

    Stokasimov, Ema; Rubenstein, Peter A.

    2009-01-01

    Actin can exist in multiple conformations necessary for normal function. Actin isoforms, although highly conserved in sequence, exhibit different biochemical properties and cellular roles. We used amide proton hydrogen/deuterium (HD) exchange detected by mass spectrometry to analyze conformational differences between Saccharomyces cerevisiae and muscle actins in the G and F forms to gain insight into these differences. We also utilized HD exchange to study interdomain and allosteric communication in yeast-muscle hybrid actins to better understand the conformational dynamics of actin. Areas showing differences in HD exchange between G- and F-actins are areas of intermonomer contacts, consistent with the current filament models. Our results showed greater exchange for yeast G-actin compared with muscle actin in the barbed end pivot region and areas in subdomains 1 and 2 and for F-actin in monomer-monomer contact areas. These results suggest greater flexibility of the yeast actin monomer and filament compared with muscle actin. For hybrid G-actins, the muscle-like and yeastlike parts of the molecule generally showed exchange characteristics resembling their parent actins. A few exceptions were a peptide on top of subdomain 2 and the pivot region between subdomains 1 and 3 with muscle actin-like exchange characteristics although the areas were yeastlike. These results demonstrate that there is cross-talk between subdomains 1 and 2 and the large and small domains. Hybrid F-actin data showing greater exchange compared with both yeast and muscle actins are consistent with mismatched yeast-muscle interfaces resulting in decreased stability of the hybrid filament contacts. PMID:19605362

  2. Probing conformational changes in rhodopsin using hydrogen-deuterium exchange coupled to mass spectrometry.

    Science.gov (United States)

    Orban, Tivadar; Tsybovsky, Yaroslav

    2015-01-01

    Hydrogen-deuterium exchange coupled to mass spectrometry is a powerful tool to evaluate changes in protein conformation between two or more states. Here, we describe a complete methodology that can be used to assess conformational changes in rhodopsin accompanying its transition from the inactive to activated state upon light exposure. This approach may be employed to investigate the structure and conformational changes of various membrane proteins.

  3. On Decentralization and Life Satisfaction

    DEFF Research Database (Denmark)

    Bjørnskov, Christian; Dreher, Axel; Fischer, Justina A.V.

    2008-01-01

    We empirically analyze the impact of fiscal and political decentralization on subjective well-being in a cross-section of 60,000 individuals from 66 countries. More spending or revenue decentralization raises well-being while greater local autonomy is beneficial only via government consumption sp...

  4. A contingency approach to decentralization

    NARCIS (Netherlands)

    Fleurke, F.; Hulst, J.R.

    2006-01-01

    After decades of centralization, in 1980 the central government of the Netherlands embarked upon an ambitious project to decentralize the administrative system. It proclaimed a series of general decentralization measures that aimed to improve the performance of the administrative system and to boost

  5. High-sensitivity mass spectrometry for imaging subunit interactions: hydrogen/deuterium exchange.

    Science.gov (United States)

    Lanman, Jason; Prevelige, Peter E

    2004-04-01

    In recent years, advances in mass spectrometry have provided unprecedented knowledge of protein expression within cells. It has become apparent that many proteins function as macromolecular complexes. Structural genomics programs are determining the fold of these proteins at an increasing rate and electron microscopic tomography potentially provides a means to determine the location of these complexes within the cell. A complete understanding of the molecular mechanism of these proteins requires detailed information on the interactions and dynamics within the complex. Recent advances in mass spectrometry now make it possible to use hydrogen/deuterium exchange to detect intersubunit interfaces and dynamics within supramolecular complexes.

  6. Analysis for corruption and decentralization (Case study: earlier decentralization era in Indonesia

    Directory of Open Access Journals (Sweden)

    Joko Tri Haryanto

    2017-06-01

    Full Text Available Abstrak. Di banyak negara, hubungan antara desentralisasi pemerintah dan tingkat ekstraksi sewa oleh pihak swasta merupakan elemen penting dalam perdebatan baru pada desain institusional. Topik korupsi secara aktif, terbuka dan diperdebatkan di Indonesia oleh Pemerintah, mitra pembangunan, dan kelompok berbasis luas dari para pemimpin politik dan masyarakat sipil yang terlibat dalam pertemuan dan pertukaran setiap hari. Dalam perdebatan tentang korupsi banyak perhatian diarahkan untuk peran gaji sektor publik, terutama di era desentralisasi. Berdasarkan fenomena ini, penulis ingin menganalisis hubungan antara korupsi dan desentralisasi. Menggunakan OSL model, kita dapat menemukan hubungan positif yang sangat kuat dan konsisten antara dua variabel di seluruh sampel dari daerah, sehingga memberikan beberapa dukungan untuk teori desentralisasi yang menekankan manfaat. Asosiasi ini adalah kuat untuk mengendalikan berbagai kemungkinan potensial dari upaya menghilangkan sebagian variabel serta bias endogenitas. Kata Kunci: Korupsi, Desentralisasi, OSL Model   Abstract. In many countries, relationship between decentralization of government activities and the extent of rent extraction by private parties is an important element in the recent debate on institutional design. The topic of corruption was actively, openly and debated in Indonesia by government, its development partners, and a broadly based group of political and civil society leaders are engaged in meetings and exchange on a daily basis. In the ongoing debate on corruption a lot of attention is paid to the role of public sector salaries, particularly in the decentralization era. Based on this phenomenon, the authors want to analyze the relationship between corruption and decentralization. Using OSL model, we can find a very strong and consistent positive association between the two variables across a sample of region, thereby providing some support for theories of decentralization that

  7. On the vertical exchange of heat, mass and momentum over complex, mountainous terrain

    Directory of Open Access Journals (Sweden)

    Mathias Walter Rotach

    2015-12-01

    Full Text Available The role of the atmospheric boundary layer (ABL in the atmosphere-climate system is the exchange of heat, mass and momentum between ‘the earth’s surface’ and the atmosphere. Traditionally, it is understood that turbulent transport is responsible for this exchange and hence the understanding and physical description of the turbulence structure of the boundary layer is key to assess the effectiveness of earth-atmosphere exchange. This understanding is rooted in the (implicit assumption of a scale separation or spectral gap between turbulence and mean atmospheric motions, which in turn leads to the assumption of a horizontally homogeneous and flat (HHF surface as a reference, for which both physical understanding and model parameterizations have successfully been developed over the years. Over mountainous terrain, however, the ABL is generically inhomogeneous due to both thermal (radiative and dynamic forcing. This inhomogeneity leads to meso-scale and even sub-meso-scale flows such as slope and valley winds or wake effects. It is argued here that these (submeso-scale motions can significantly contribute to the vertical structure of the boundary layer and hence vertical exchange of heat and mass between the surface and the atmosphere. If model grid resolution is not high enough the latter will have to be parameterized (in a similar fashion as gravity wave drag parameterizations take into account the momentum transport due to gravity waves in large-scale models. In this contribution we summarize the available evidence of the contribution of (submeso-scale motions to vertical exchange in mountainous terrain from observational and numerical modeling studies. In particular, a number of recent simulation studies using idealized topography will be summarized and put into perspective – so as to identify possible limitations and areas of necessary future research.

  8. Mathematical model development of heat and mass exchange processes in the outdoor swimming pool

    Directory of Open Access Journals (Sweden)

    M. V. Shaptala

    2014-12-01

    Full Text Available Purpose. Currently exploitation of outdoor swimming pools is often not cost-effective and, despite of their relevance, such pools are closed in large quantities. At this time there is no the whole mathematical model which would allow assessing qualitatively the effect of energy-saving measures. The aim of this work is to develop a mathematical model of heat and mass exchange processes for calculating basic heat and mass losses that occur during its exploitation. Methodology. The method for determination of heat and mass loses based on the theory of similarity criteria equations is used. Findings. The main types of heat and mass losses of outdoor pool were analyzed. The most significant types were allocated and mathematically described. Namely: by evaporation of water from the surface of the pool, by natural and forced convection, by radiation to the environment, heat consumption for water heating. Originality. The mathematical model of heat and mass exchange process of the outdoor swimming pool was developed, which allows calculating the basic heat and mass loses that occur during its exploitation. Practical value. The method of determining heat and mass loses of outdoor swimming pool as a software system was developed and implemented. It is based on the mathematical model proposed by the authors. This method can be used for the conceptual design of energy-efficient structures of outdoor pools, to assess their use of energy-intensive and selecting the optimum energy-saving measures. A further step in research in this area is the experimental validation of the method of calculation of heat losses in outdoor swimming pools with its use as an example the pool of Dnipropetrovsk National University of Railway Transport named after Academician V. Lazaryan. The outdoor pool, with water heating- up from the boiler room of the university, is operated year-round.

  9. Mathematical Modeling of Non-Fickian Diffusional Mass Exchange of Radioactive Contaminants in Geological Disposal Formations

    Directory of Open Access Journals (Sweden)

    Anna Suzuki

    2018-01-01

    Full Text Available Deep geological repositories for nuclear wastes consist of both engineered and natural geologic barriers to isolate the radioactive material from the human environment. Inappropriate repositories of nuclear waste would cause severe contamination to nearby aquifers. In this complex environment, mass transport of radioactive contaminants displays anomalous behaviors and often produces power-law tails in breakthrough curves due to spatial heterogeneities in fractured rocks, velocity dispersion, adsorption, and decay of contaminants, which requires more sophisticated models beyond the typical advection-dispersion equation. In this paper, accounting for the mass exchange between a fracture and a porous matrix of complex geometry, the universal equation of mass transport within a fracture is derived. This equation represents the generalization of the previously used models and accounts for anomalous mass exchange between a fracture and porous blocks through the introduction of the integral term of convolution type and fractional derivatives. This equation can be applied for the variety of processes taking place in the complex fractured porous medium, including the transport of radioactive elements. The Laplace transform method was used to obtain the solution of the fractional diffusion equation with a time-dependent source of radioactive contaminant.

  10. Probing protein interactions with hydrogen/deuterium exchange and mass spectrometry—A review

    International Nuclear Information System (INIS)

    Percy, Andrew J.; Rey, Martial; Burns, Kyle M.; Schriemer, David C.

    2012-01-01

    Highlights: ► Protein chemistry generates mass shifts useful for structure–function studies. ► H/DX supports a powerful mass shift method for protein interaction analysis. ► H/DX mass shifts are useful for determining binding data (K d , off-rates). ► Improved H/DX–MS workflows can accommodate complex protein systems. - Abstract: Assessing the functional outcome of protein interactions in structural terms is a goal of structural biology, however most techniques have a limited capacity for making structure–function determinations with both high resolution and high throughput. Mass spectrometry can be applied as a reader of protein chemistries in order to fill this void, and enable methodologies whereby protein structure–function determinations may be made on a proteome-wide level. Protein hydrogen/deuterium exchange (H/DX) offers a chemical labeling strategy suitable for tracking changes in “dynamic topography” and thus represents a powerful means of monitoring protein structure–function relationships. This review presents the exchange method in the context of interaction analysis. Applications involving interface detection, quantitation of binding, and conformational responses to ligation are discussed, and commentary on recent analytical developments is provided.

  11. Mechanism of framework oxygen exchange in Fe-zeolites: a combined DFT and mass spectrometry study.

    Science.gov (United States)

    Andrikopoulos, Prokopis C; Sobalik, Zdenek; Novakova, Jana; Sazama, Petr; Sklenak, Stepan

    2013-02-25

    The role of framework oxygen atoms in N(2)O decomposition [N(2)O(g)→N(2)(g) and 1/2O(2)(g)] over Fe-ferrierite is investigated employing a combined experimental (N(2)(18)O decomposition in batch experiments followed by mass spectroscopy measurements) and theoretical (density functional theory calculations) approach. The occurrence of the isotope exchange indicates that framework oxygen atoms are involved in the N(2)O decomposition catalyzed by Fe-ferrierite. Our study, using an Fe-ferrierite sample with iron exclusively present as Fe(II) cations accommodated in the cationic sites, shows that the mobility of framework oxygen atoms in the temperature range: 553 to 593 K is limited to the four framework oxygen atoms of the two AlO(4)(-) tetrahedra forming cationic sites that accomodate Fe(II). They exchange with the Fe extra-framework (18)O atom originating from the decomposed N(2)(18)O. We found, using DFT calculations, that O(2) molecules facilitate the oxygen exchange. However, the corresponding calculated energy barrier of 87 kcal  mol(-1) is still very high and it is higher than the assumed experimental value based on the occurrence of the sluggish oxygen exchange at 553 K. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Structural changes of ultrasonicated bovine serum albumin revealed by hydrogen-deuterium exchange and mass spectrometry.

    Science.gov (United States)

    Zhang, Qiuting; Tu, Zongcai; Wang, Hui; Huang, Xiaoqin; Sha, Xiaomei; Xiao, Hui

    2014-11-01

    The structural changes of bovine serum albumin (BSA) under high-intensity ultrasonication were investigated by fluorescence spectroscopy and mass spectrometry. Evidence for the ultrasonication-induced conformational changes of BSA was provided by the intensity changes and maximum-wavelength shift in fluorescence spectrometry. Matrix-assisted laser desorption-ionization time-of-flight mass spectroscopy (MALDI-TOF MS) revealed the increased intensity of the peak at the charge state +5 and a newly emerged peak at charge state +6, indicating that the protein became unfolded after ultrasonication. Prevalent unfolding of BSA after ultrasonication was revealed by hydrogen-deuterium exchange coupled with mass spectrometry (HDX-MS). Increased intensity and duration of ultrasonication further promoted the unfolding of the protein. The unfolding induced by ultrasonication goes through an intermediate state similar to that induced by a low concentration of denaturant.

  13. Hydrogen deuterium exchange mass spectrometry in biopharmaceutical discovery and development – A review

    Energy Technology Data Exchange (ETDEWEB)

    Deng, Bin, E-mail: dengbin@yorku.ca [Chemistry Department, York University, 4700 Keele Street, Toronto, ON, M3J 1P3 (Canada); The Centre for Research in Mass Spectrometry, York University, Toronto, ON, M3J1P3 (Canada); Lento, Cristina, E-mail: clento@yorku.ca [Chemistry Department, York University, 4700 Keele Street, Toronto, ON, M3J 1P3 (Canada); The Centre for Research in Mass Spectrometry, York University, Toronto, ON, M3J1P3 (Canada); Wilson, Derek J., E-mail: dkwilson@yorku.ca [Chemistry Department, York University, 4700 Keele Street, Toronto, ON, M3J 1P3 (Canada); The Centre for Research in Mass Spectrometry, York University, Toronto, ON, M3J1P3 (Canada)

    2016-10-12

    Protein therapeutics have emerged as a major class of biopharmaceuticals over the past several decades, a trend that has motivated the advancement of bioanalytical technologies for protein therapeutic characterization. Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a powerful and sensitive technique that can probe the higher order structure of proteins and has been used in the assessment and development of monoclonal antibodies (mAbs), antibody-drug conjugates (ADCs) and biosimilar antibodies. It has also been used to quantify protein-ligand, protein-receptor and other protein-protein interactions involved in signaling pathways. In manufacturing and development, HDX-MS can validate storage formulations and manufacturing processes for various biotherapeutics. Currently, HDX-MS is being refined to provide additional coverage, sensitivity and structural specificity and implemented on the millisecond timescale to reveal residual structure and dynamics in disordered domains and intrinsically disordered proteins. - Highlights: • The pharmaceuticals industry is increasingly shifting to protein therapeutics. • Hydrogen deuterium exchange mass spectrometry is uniquely well suited to support biopharmaceutical development. • Applications for hydrogen deuterium exchange span drug discovery, development and manufacturing. • Future developments will allow improved sensitivity, structural resolution and a broader range of dynamics to be monitored.

  14. Improved protein hydrogen/deuterium exchange mass spectrometry platform with fully automated data processing.

    Science.gov (United States)

    Zhang, Zhongqi; Zhang, Aming; Xiao, Gang

    2012-06-05

    Protein hydrogen/deuterium exchange (HDX) followed by protease digestion and mass spectrometric (MS) analysis is accepted as a standard method for studying protein conformation and conformational dynamics. In this article, an improved HDX MS platform with fully automated data processing is described. The platform significantly reduces systematic and random errors in the measurement by introducing two types of corrections in HDX data analysis. First, a mixture of short peptides with fast HDX rates is introduced as internal standards to adjust the variations in the extent of back exchange from run to run. Second, a designed unique peptide (PPPI) with slow intrinsic HDX rate is employed as another internal standard to reflect the possible differences in protein intrinsic HDX rates when protein conformations at different solution conditions are compared. HDX data processing is achieved with a comprehensive HDX model to simulate the deuterium labeling and back exchange process. The HDX model is implemented into the in-house developed software MassAnalyzer and enables fully unattended analysis of the entire protein HDX MS data set starting from ion detection and peptide identification to final processed HDX output, typically within 1 day. The final output of the automated data processing is a set (or the average) of the most possible protection factors for each backbone amide hydrogen. The utility of the HDX MS platform is demonstrated by exploring the conformational transition of a monoclonal antibody by increasing concentrations of guanidine.

  15. Hydrogen deuterium exchange mass spectrometry in biopharmaceutical discovery and development – A review

    International Nuclear Information System (INIS)

    Deng, Bin; Lento, Cristina; Wilson, Derek J.

    2016-01-01

    Protein therapeutics have emerged as a major class of biopharmaceuticals over the past several decades, a trend that has motivated the advancement of bioanalytical technologies for protein therapeutic characterization. Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a powerful and sensitive technique that can probe the higher order structure of proteins and has been used in the assessment and development of monoclonal antibodies (mAbs), antibody-drug conjugates (ADCs) and biosimilar antibodies. It has also been used to quantify protein-ligand, protein-receptor and other protein-protein interactions involved in signaling pathways. In manufacturing and development, HDX-MS can validate storage formulations and manufacturing processes for various biotherapeutics. Currently, HDX-MS is being refined to provide additional coverage, sensitivity and structural specificity and implemented on the millisecond timescale to reveal residual structure and dynamics in disordered domains and intrinsically disordered proteins. - Highlights: • The pharmaceuticals industry is increasingly shifting to protein therapeutics. • Hydrogen deuterium exchange mass spectrometry is uniquely well suited to support biopharmaceutical development. • Applications for hydrogen deuterium exchange span drug discovery, development and manufacturing. • Future developments will allow improved sensitivity, structural resolution and a broader range of dynamics to be monitored.

  16. Decentring the Creative Self

    DEFF Research Database (Denmark)

    Glaveanu, Vlad Petre; Lubart, Todd

    2014-01-01

    Since its inception, the psychology of creativity has been concerned primarily with the study of individual creators. In contrast, this research is dedicated to an exploration of (a) who has a significant impact on a creative professional's activity and (b) what the contribution is that others make...... to themes depicting the interaction between these different others and the creator. Findings reveal both similarities and differences across the five domains in terms of the specific contribution of others to the creative process. Social interactions play a key formative, regulatory, motivational...... and informational role in relation to creative work. From ‘internalized’ to ‘distant’, other people are an integral part of the equation of creativity calling for a de-centring of the creative self and its re-centring in a social space of actions and interactions....

  17. Simple atmospheric hydrogen/deuterium exchange method for enumeration of labile hydrogens by electrospray ionization mass spectrometry.

    Science.gov (United States)

    Kostyukevich, Yury; Kononikhin, Alexey; Popov, Igor; Nikolaev, Eugene

    2013-06-04

    A simple method for hydrogen/deuterium exchange in a standard electrospray (ESI) ionization source is presented. In this method, a D₂O droplet is placed between the ESI needle and the entrance of the mass spectrometer and thus saturation of the atmosphere with deuterated vapor in the ESI region is achieved. It was shown that full exchange of up to 23 labile acidic hydrogens with a minimal back exchange with the surrounding atmospheric water can be performed by this method.

  18. Aspartic protease nepenthesin-1 as a tool for digestion in hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Kadek, Alan; Mrazek, Hynek; Halada, Petr; Rey, Martial; Schriemer, David C; Man, Petr

    2014-05-06

    Hydrogen/deuterium exchange coupled to mass spectrometry (HXMS) utilizes enzymatic digestion of proteins to localize the information about altered exchange patterns in protein structure. The ability of the protease to produce small peptides and overlapping fragments and provide sufficient coverage of the protein sequence is essential for localizing regions of interest. Recently, it was shown that there is an interesting group of proteolytic enzymes from carnivorous pitcher plants of the genus Nepenthes. In this report, we describe successful immobilization and the use of one of these enzymes, nepenthesin-1, in HXMS workflow. In contrast to pepsin, it has different cleavage specificities, and despite its high inherent susceptibility to reducing and denaturing agents, it is very stable upon immobilization and withstands even high concentration of guanidine hydrochloride and reducing agents. We show that denaturing agents can alter digestion by reducing protease activity and/or substrate solubility, and additionally, they influence the trapping of proteolytic peptides onto the reversed phase resin.

  19. Analysis of Overlapped and Noisy Hydrogen/Deuterium Exchange Mass Spectra

    Science.gov (United States)

    Guttman, Miklos; Weis, David D.; Engen, John R.; Lee, Kelly K.

    2013-12-01

    Noisy and overlapped mass spectrometry data hinder the sequence coverage that can be obtained from hydrogen deuterium exchange analysis, and places a limit on the complexity of the samples that can be studied by this technique. Advances in instrumentation have addressed these limits, but as the complexity of the biological samples under investigation increases, these problems are re-encountered. Here we describe the use of binomial distribution fitting with asymmetric linear squares regression for calculating the accurate deuterium content for mass envelopes of low signal or that contain significant overlap. The approach is demonstrated with a test data set of HIV Env gp140 wherein inclusion of the new analysis regime resulted in obtaining exchange data for 42 additional peptides, improving the sequence coverage by 11 %. At the same time, the precision of deuterium uptake measurements was improved for nearly every peptide examined. The improved processing algorithms also provide an efficient method for deconvolution of bimodal mass envelopes and EX1 kinetic signatures. All these functions and visualization tools have been implemented in the new version of the freely available software, HX-Express v2.

  20. An overview of hydrogen deuterium exchange mass spectrometry (HDX-MS) in drug discovery.

    Science.gov (United States)

    Masson, Glenn R; Jenkins, Meredith L; Burke, John E

    2017-10-01

    Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a powerful methodology to study protein dynamics, protein folding, protein-protein interactions, and protein small molecule interactions. The development of novel methodologies and technical advancements in mass spectrometers has greatly expanded the accessibility and acceptance of this technique within both academia and industry. Areas covered: This review examines the theoretical basis of how amide exchange occurs, how different mass spectrometer approaches can be used for HDX-MS experiments, as well as the use of HDX-MS in drug development, specifically focusing on how HDX-MS is used to characterize bio-therapeutics, and its use in examining protein-protein and protein small molecule interactions. Expert opinion: HDX-MS has been widely accepted within the pharmaceutical industry for the characterization of bio-therapeutics as well as in the mapping of antibody drug epitopes. However, there is room for this technique to be more widely used in the drug discovery process. This is particularly true in the use of HDX-MS as a complement to other high-resolution structural approaches, as well as in the development of small molecule therapeutics that can target both active-site and allosteric binding sites.

  1. Advantages of isotopic depletion of proteins for hydrogen/deuterium exchange experiments monitored by mass spectrometry.

    Science.gov (United States)

    Bou-Assaf, George M; Chamoun, Jean E; Emmett, Mark R; Fajer, Piotr G; Marshall, Alan G

    2010-04-15

    Solution-phase hydrogen/deuterium exchange (HDX) monitored by mass spectrometry is an excellent tool to study protein-protein interactions and conformational changes in biological systems, especially when traditional methods such as X-ray crystallography or nuclear magnetic resonance are not feasible. Peak overlap among the dozens of proteolytic fragments (including those from autolysis of the protease) can be severe, due to high protein molecular weight(s) and the broad isotopic distributions due to multiple deuterations of many peptides. In addition, different subunits of a protein complex can yield isomeric proteolytic fragments. Here, we show that depletion of (13)C and/or (15)N for one or more protein subunits of a complex can greatly simplify the mass spectra, increase the signal-to-noise ratio of the depleted fragment ions, and remove ambiguity in assignment of the m/z values to the correct isomeric peptides. Specifically, it becomes possible to monitor the exchange progress for two isobaric fragments originating from two or more different subunits within the complex, without having to resort to tandem mass spectrometry techniques that can lead to deuterium scrambling in the gas phase. Finally, because the isotopic distribution for a small to medium-size peptide is essentially just the monoisotopic species ((12)C(c)(1)H(h)(14)N(n)(16)O(o)(32)S(s)), it is not necessary to deconvolve the natural abundance distribution for each partially deuterated peptide during HDX data reduction.

  2. Analysis of overlapped and noisy hydrogen/deuterium exchange mass spectra.

    Science.gov (United States)

    Guttman, Miklos; Weis, David D; Engen, John R; Lee, Kelly K

    2013-12-01

    Noisy and overlapped mass spectrometry data hinder the sequence coverage that can be obtained from hydrogen deuterium exchange analysis, and places a limit on the complexity of the samples that can be studied by this technique. Advances in instrumentation have addressed these limits, but as the complexity of the biological samples under investigation increases, these problems are re-encountered. Here we describe the use of binomial distribution fitting with asymmetric linear squares regression for calculating the accurate deuterium content for mass envelopes of low signal or that contain significant overlap. The approach is demonstrated with a test data set of HIV Env gp140 wherein inclusion of the new analysis regime resulted in obtaining exchange data for 42 additional peptides, improving the sequence coverage by 11%. At the same time, the precision of deuterium uptake measurements was improved for nearly every peptide examined. The improved processing algorithms also provide an efficient method for deconvolution of bimodal mass envelopes and EX1 kinetic signatures. All these functions and visualization tools have been implemented in the new version of the freely available software, HX-Express v2.

  3. Structural analysis of the interleukin-8/glycosaminoglycan interactions by amide hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Hofmann, Tommy; Samsonov, Sergey A; Pichert, Annelie; Lemmnitzer, Katharina; Schiller, Jürgen; Huster, Daniel; Pisabarro, M Teresa; von Bergen, Martin; Kalkhof, Stefan

    2015-11-01

    The recruitment of different chemokines and growth factors by glycosaminoglycans (GAGs) such as chondroitin sulfate or hyaluronan plays a critical role in wound healing processes. Thus, there is a special interest in the design of artificial extracellular matrices with improved properties concerning GAG interaction with common regulating proteins. In this study, amide hydrogen/deuterium (H/D) exchange mass spectrometry (HDX MS) combined with molecular modeling and docking experiments was used to obtain structural models of proinflammatory chemokine interleukin-8 (IL-8) in complex with hexameric chondroitin sulfate. Experiments on the intact protein showed a difference in deuterium labeling of IL-8 due to chondroitin sulfate binding. The extent of deuteration was reduced from 24% to 13% after 2 min exchange time, which corresponds to a reduced exchange of approximately 10 backbone amides. By local HDX MS experiments, H/D exchange information on the complete sequence of IL-8 could be obtained. A significantly reduced H/D exchange, especially of the C-terminal α-helical region comprising amino acids 70-77 and to the loop comprising amino acids 27-29 was observed in the presence of chondroitin sulfate. HDX MS data were used to model the IL-8/chondroitin sulfate complex. The binding interface of IL-8 and chondroitin sulfate determined this way correlated excellently with the corresponding NMR based atomistic model previously published. Our results demonstrate that HDX-MS in combination with molecular modeling is a valuable approach for the analysis of protein/GAG complexes at physiological pH, temperature, and salt concentration. The fact that HDX-MS requires only micrograms of protein and GAGs makes it a very promising technique to address protein-GAG interactions. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Guanine nucleotide induced conformational change of Cdc42 revealed by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Yang, Sheng-Wei; Ting, Hsiu-Chi; Lo, Yi-Ting; Wu, Ting-Yuan; Huang, Hung-Wei; Yang, Chia-Jung; Chan, Jui-Fen Riva; Chuang, Min-Chieh; Hsu, Yuan-Hao Howard

    2016-01-01

    Cdc42 regulates pathways related to cell division. Dysregulation of Cdc42 can lead to cancer, cardiovascular diseases and neurodegenerative diseases. GTP induced activation mechanism plays an important role in the activity and biological functions of Cdc42. P-loop, Switch I and Switch II are critical regions modulating the enzymatic activity of Cdc42. We applied amide hydrogen/deuterium exchange coupled with liquid chromatography mass spectrometry (HDXMS) to investigate the dynamic changes of apo-Cdc42 after GDP, GTP and GMP-PCP binding. The natural substrate GTP induced significant decreases of deuteration in P-loop and Switch II, moderate changes of deuteration in Switch I and significant changes of deuteration in the α7 helix, a region far away from the active site. GTP binding induced similar effects on H/D exchange to its non-hydrolysable analog, GMP-PCP. HDXMS results indicate that GTP binding blocked the solvent accessibility in the active site leading to the decrease of H/D exchange rate surrounding the active site, and further triggered a conformational change resulting in the drastic decrease of H/D exchange rate at the remote α7 helix. Comparing the deuteration levels in three activation states of apo-Cdc42, Cdc42-GDP and Cdc42-GMP-PCP, the apo-Cdc42 has the most flexible structure, which can be stabilized by guanine nucleotide binding. The rates of H/D exchange of Cdc42-GDP are between the GMP-PCP-bound and the apo form, but more closely to the GMP-PCP-bound form. Our results show that the activation of Cdc42 is a process of conformational changes involved with P-loop, Switch II and α7 helix for structural stabilization. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Analysis of phosphoinositide 3-kinase inhibitors by bottom-up electron-transfer dissociation hydrogen/deuterium exchange mass spectrometry

    OpenAIRE

    Masson, Glenn R.; Maslen, Sarah L.; Williams, Roger L.

    2017-01-01

    Until recently, one of the major limitations of hydrogen/deuterium exchange mass spectrometry (HDX-MS) was the peptide-level resolution afforded by proteolytic digestion. This limitation can be selectively overcome through the use of electron-transfer dissociation to fragment peptides in a manner that allows the retention of the deuterium signal to produce hydrogen/deuterium exchange tandem mass spectrometry (HDX-MS/MS). Here, we describe the application of HDX-MS/MS to structurally screen in...

  6. Decentralization Calls for Internal Audits.

    Science.gov (United States)

    DiCello, Jim

    1995-01-01

    Outlines internal-auditing strategies necessitated by decentralization. Describes the following areas of concern: the student activities account, student attendance, and funding delegated to the site level. Guidelines for conducting an internal audit are also included. (LMI)

  7. Mathematical modeling of water mass balance for proton exchange membrane fuel cell

    International Nuclear Information System (INIS)

    Wan Ramli Wan Daud; Kamaruzzaman Sopian; Jaafar Sahari; Nik Suhaimi Mat Hassan

    2006-01-01

    Gas and water management are key to achieving good performance from a proton exchange membrane fuel cell (PEMFC) stack. Water plays a critical role in PEMFC. The proton conductivity is increase with the water content. In order to achieve enough hydration, water is normally introduced into the cell externally by a variety of methods such as liquid injection, steam introduction, and humidification of reactants by passing them through humidifiers before entering the cell. In this paper, mathematical modeling of water mass balance for PEMFC at anode and cathode side are proposed by using external humidification and assume that steady state, constant pressure, constant temperature and gases distribution are uniform

  8. Folding and assembly of large macromolecular complexes monitored by hydrogen-deuterium exchange and mass spectrometry

    Directory of Open Access Journals (Sweden)

    Tuma Roman

    2008-04-01

    Full Text Available Abstract Recent advances in protein mass spectrometry (MS have enabled determinations of hydrogen deuterium exchange (HDX in large macromolecular complexes. HDX-MS became a valuable tool to follow protein folding, assembly and aggregation. The methodology has a wide range of applications in biotechnology ranging from quality control for over-expressed proteins and their complexes to screening of potential ligands and inhibitors. This review provides an introduction to protein folding and assembly followed by the principles of HDX and MS detection, and concludes with selected examples of applications that might be of interest to the biotechnology community.

  9. Decentralization and Bailouts in Colombia

    OpenAIRE

    Juan José Echavarría; Carolina Rentería; Roberto Steiner

    2002-01-01

    This paper examines the decentralization process in Colombia, particularly as it relates to reforms embodied in the 1991 Constitution, and the bailouts of territorial entities that have occurred in subsequent years as a result of perverse incentive structures. The paper provides a summary of the basic features of the decentralization process in Colombia, including an analysis of revenue and expenditure assignments, the intergovernmental transfer system and recent developments in terms of terr...

  10. Decentralized Bribery and Market Participation

    OpenAIRE

    Popov, Sergey V.

    2012-01-01

    I propose a bribery model that examines decentralized bureaucratic decision-making. There are multiple stable equilibria. High levels of bribery reduce an economy's productivity because corruption suppresses small business, and reduces the total graft, even though the size of an individual bribe might increase. Decentralization prevents movement towards a Pareto-dominant equilibrium. Anticorruption efforts, even temporary ones, might be useful to improve participation, if they lower the bribe...

  11. Hydrogen-deuterium exchange studies utilizing a thermospray mass spectrometer interface

    International Nuclear Information System (INIS)

    Siegel, M.M.

    1988-01-01

    A routine and efficient technique was developed for exchanging acidic hydrogen for deuterium, utilizing a thermospray mass spectrometer interface, to produce deuterium-enriched deuteron-adduct molecular and fragment ions. Samples dissolved in solvents containing acidic deuterium atoms (D 2 O/trifluoroacetic acid-d 1 or ammonium acetate-d 4 ) were introduced via slug injection into the thermospray interface sandwiched between slugs of the deuteriated solvent. At all other times, the carrier solvent system introduced into the mass spectrometer consisted of a non-deuterium-enriched mixture. Deuterium enrichments achieved were greater than 95%. Deuterium-enriched deuteron-adduct molecular and fragment ions enabled the determination of the number of acidic hydrogens and aided in structural elucidation of model compounds of pharmaceutical interest

  12. Spatially resolved protein hydrogen exchange measured by subzero-cooled chip-based nanoelectrospray ionization tandem mass spectrometry

    DEFF Research Database (Denmark)

    Amon, Sabine; Trelle, Morten B; Jensen, Ole N

    2012-01-01

    Mass spectrometry has become a valuable method for studying structural dynamics of proteins in solution by measuring their backbone amide hydrogen/deuterium exchange (HDX) kinetics. In a typical exchange experiment one or more proteins are incubated in deuterated buffer at physiological conditions....... After a given period of deuteration, the exchange reaction is quenched by acidification (pH 2.5) and cooling (0 °C) and the deuterated protein (or a digest thereof) is analyzed by mass spectrometry. The unavoidable loss of deuterium (back-exchange) that occurs under quench conditions is undesired......, only 4% and 6% deuterium loss for fully deuterated ubiquitin and β(2)-microglobulin were observed after 10 min of back-exchange. The practical value of our subzero-cooled setup for top-down fragmentation HDX analyses is demonstrated by electron-transfer dissociation of ubiquitin ions under carefully...

  13. Conformational changes in oxidatively stressed monoclonal antibodies studied by hydrogen exchange mass spectrometry

    Science.gov (United States)

    Burkitt, William; Domann, Paula; O'Connor, Gavin

    2010-01-01

    Oxidation of methionine residues in biopharmaceuticals is a common and often unwanted modification that frequently occurs during their manufacture and storage. It often results in a lack of stability and biological function of the product, necessitating continuous testing for the modification throughout the product shelf life. A major class of biopharmaceutical products are monoclonal antibodies (mAbs), however, techniques for their detailed structural analysis have until recently been limited. Hydrogen/deuterium exchange mass spectrometry (HXMS) has recently been successfully applied to the analysis of mAbs. Here we used HXMS to identify and localise the structural changes that occurred in a mAb (IgG1) after accelerated oxidative stress. Structural alterations in a number of segments of the Fc region were observed and these related to oxidation of methionine residues. These included a large change in the hydrogen exchange profile of residues 247–253 of the heavy chain, while smaller changes in hydrogen exchange profile were identified for peptides that contained residues in the interface of the CH2 and CH3 domains. PMID:20162626

  14. Real-time hydrogen/deuterium exchange kinetics via supercharged electrospray ionization tandem mass spectrometry.

    Science.gov (United States)

    Sterling, Harry J; Williams, Evan R

    2010-11-01

    Amide hydrogen/deuterium exchange (HDX) rate constants of bovine ubiquitin in an ammonium acetate solution containing 1% of the electrospray ionization (ESI) "supercharging" reagent m-nitrobenzyl alcohol (m-NBA) were obtained using top-down, electron transfer dissociation (ETD) tandem mass spectrometry (MS). The supercharging reagent replaces the acid and temperature "quench" step in the conventional MS approach to HDX experiments by causing rapid protein denaturation to occur in the ESI droplet. The higher charge state ions that are produced with m-NBA are more unfolded, as measured by ion mobility, and result in higher fragmentation efficiency and higher sequence coverage with ETD. Single amino acid resolution was obtained for 44 of 72 exchangeable amide sites, and summed kinetic data were obtained for regions of the protein where adjacent fragment ions were not observed, resulting in an overall spatial resolution of 1.3 residues. Comparison of these results with previous values from NMR indicates that the supercharging reagent does not cause significant structural changes to the protein in the initial ESI solution and that scrambling or back-exchange is minimal. This new method for top-down HDX-MS enables real-time kinetic data measurements under physiological conditions, similar to those obtained using NMR, with comparable spatial resolution and significantly better sensitivity.

  15. Removal of chlorofluorocarbons by increased mass exchange between the stratosphere and troposphere in a changing climate.

    Science.gov (United States)

    Butchart, N; Scaife, A A

    2001-04-12

    Chlorofluorocarbons (CFCs), along with bromine compounds, have been unequivocally identified as being responsible for most of the anthropogenic destruction of stratospheric ozone. With curbs on emissions of these substances, the recovery of the ozone layer will depend on their removal from the atmosphere. As CFCs have no significant tropospheric removal process, but are rapidly photolysed above the lower stratosphere, the timescale for their removal is set mainly by the rate at which air is transported from the troposphere into the stratosphere. Using a global climate model we predict that, in response to the projected changes in greenhouse-gas concentrations during the first half of the twenty-first century, this rate of mass exchange will increase by 3% per decade. This increase is due to more vigorous extra-tropical planetary waves emanating from the troposphere. We estimate that this increase in mass exchange will accelerate the removal of CFCs to an extent that recovery to levels currently predicted for 2050 and 2080 will occur 5 and 10 years earlier, respectively.

  16. Localizing Carbohydrate Binding Sites in Proteins Using Hydrogen/Deuterium Exchange Mass Spectrometry

    Science.gov (United States)

    Zhang, Jingjing; Kitova, Elena N.; Li, Jun; Eugenio, Luiz; Ng, Kenneth; Klassen, John S.

    2016-01-01

    The application of hydrogen/deuterium exchange mass spectrometry (HDX-MS) to localize ligand binding sites in carbohydrate-binding proteins is described. Proteins from three bacterial toxins, the B subunit homopentamers of Cholera toxin and Shiga toxin type 1 and a fragment of Clostridium difficile toxin A, and their interactions with native carbohydrate receptors, GM1 pentasaccharides (β-Gal-(1→3)-β-GalNAc-(1→4)[α-Neu5Ac-(2→3)]-β-Gal-(1→4)-Glc), Pk trisaccharide (α-Gal-(1→4)-β-Gal-(1→4)-Glc) and CD-grease (α-Gal-(1→3)-β-Gal-(1→4)-β-GlcNAcO(CH2)8CO2CH3), respectively, served as model systems for this study. Comparison of the differences in deuterium uptake for peptic peptides produced in the absence and presence of ligand revealed regions of the proteins that are protected against deuterium exchange upon ligand binding. Notably, protected regions generally coincide with the carbohydrate binding sites identified by X-ray crystallography. However, ligand binding can also result in increased deuterium exchange in other parts of the protein, presumably through allosteric effects. Overall, the results of this study suggest that HDX-MS can serve as a useful tool for localizing the ligand binding sites in carbohydrate-binding proteins. However, a detailed interpretation of the changes in deuterium exchange upon ligand binding can be challenging because of the presence of ligand-induced changes in protein structure and dynamics.

  17. Conformational analysis of g protein-coupled receptor signaling by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Li, Sheng; Lee, Su Youn; Chung, Ka Young

    2015-01-01

    Conformational change and protein-protein interactions are two major mechanisms of membrane protein signal transduction, including G protein-coupled receptors (GPCRs). Upon agonist binding, GPCRs change conformation, resulting in interaction with downstream signaling molecules such as G proteins. To understand the precise signaling mechanism, studies have investigated the structural mechanism of GPCR signaling using X-ray crystallography, nuclear magnetic resonance (NMR), or electron paramagnetic resonance. In addition to these techniques, hydrogen/deuterium exchange mass spectrometry (HDX-MS) has recently been used in GPCR studies. HDX-MS measures the rate at which peptide amide hydrogens exchange with deuterium in the solvent. Exposed or flexible regions have higher exchange rates and excluded or ordered regions have lower exchange rates. Therefore, HDX-MS is a useful tool for studying protein-protein interfaces and conformational changes after protein activation or protein-protein interactions. Although HDX-MS does not give high-resolution structures, it analyzes protein conformations that are difficult to study with X-ray crystallography or NMR. Furthermore, conformational information from HDX-MS can help in the crystallization of X-ray crystallography by suggesting highly flexible regions. Interactions between GPCRs and downstream signaling molecules are not easily analyzed by X-ray crystallography or NMR because of the large size of the GPCR-signaling molecule complexes, hydrophobicity, and flexibility of GPCRs. HDX-MS could be useful for analyzing the conformational mechanism of GPCR signaling. In this chapter, we discuss details of HDX-MS for analyzing GPCRs using the β2AR-G protein complex as a model system. © 2015 Elsevier Inc. All rights reserved.

  18. Decentralized Monitoring and Control of Electric Power Distribution System with a use case of Community Energy Storage System

    Science.gov (United States)

    Huq, Kazi Mohammad Moyeenull

    This work focuses on the platform and framework required for developing decentralized monitoring and control applications for a distribution system. Decentralized control of Community Energy Storage (CES) devices has been considered as a use case. The first chapter gives an overview of energy management system and applications. Then the development of centralized and decentralized CES control applications are discussed. The next chapters discusses the software architecture of a typical centralized SCADAEMS platform and outlines the architecture required for a decentralized platform. Then, suitability of IEC 61850 standard for decentralized monitoring and control is investigated and an approach to developing decentralized applications in IEC 61850 enabled environment is proposed and demonstrated. Finally, CIM standard is reviewed for exchanging power system topology information for distributed control.

  19. Platform Dependencies in Bottom-up Hydrogen/Deuterium Exchange Mass Spectrometry*

    Science.gov (United States)

    Burns, Kyle M.; Rey, Martial; Baker, Charles A. H.; Schriemer, David C.

    2013-01-01

    Hydrogen-deuterium exchange mass spectrometry is an important method for protein structure-function analysis. The bottom-up approach uses protein digestion to localize deuteration to higher resolution, and the essential measurement involves centroid mass determinations on a very large set of peptides. In the course of evaluating systems for various projects, we established two (HDX-MS) platforms that consisted of a FT-MS and a high-resolution QTOF mass spectrometer, each with matched front-end fluidic systems. Digests of proteins spanning a 20–110 kDa range were deuterated to equilibrium, and figures-of-merit for a typical bottom-up (HDX-MS) experiment were compared for each platform. The Orbitrap Velos identified 64% more peptides than the 5600 QTOF, with a 42% overlap between the two systems, independent of protein size. Precision in deuterium measurements using the Orbitrap marginally exceeded that of the QTOF, depending on the Orbitrap resolution setting. However, the unique nature of FT-MS data generates situations where deuteration measurements can be inaccurate, because of destructive interference arising from mismatches in elemental mass defects. This is shown through the analysis of the peptides common to both platforms, where deuteration values can be as low as 35% of the expected values, depending on FT-MS resolution, peptide length and charge state. These findings are supported by simulations of Orbitrap transients, and highlight that caution should be exercised in deriving centroid mass values from FT transients that do not support baseline separation of the full isotopic composition. PMID:23197788

  20. Platform dependencies in bottom-up hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Burns, Kyle M; Rey, Martial; Baker, Charles A H; Schriemer, David C

    2013-02-01

    Hydrogen-deuterium exchange mass spectrometry is an important method for protein structure-function analysis. The bottom-up approach uses protein digestion to localize deuteration to higher resolution, and the essential measurement involves centroid mass determinations on a very large set of peptides. In the course of evaluating systems for various projects, we established two (HDX-MS) platforms that consisted of a FT-MS and a high-resolution QTOF mass spectrometer, each with matched front-end fluidic systems. Digests of proteins spanning a 20-110 kDa range were deuterated to equilibrium, and figures-of-merit for a typical bottom-up (HDX-MS) experiment were compared for each platform. The Orbitrap Velos identified 64% more peptides than the 5600 QTOF, with a 42% overlap between the two systems, independent of protein size. Precision in deuterium measurements using the Orbitrap marginally exceeded that of the QTOF, depending on the Orbitrap resolution setting. However, the unique nature of FT-MS data generates situations where deuteration measurements can be inaccurate, because of destructive interference arising from mismatches in elemental mass defects. This is shown through the analysis of the peptides common to both platforms, where deuteration values can be as low as 35% of the expected values, depending on FT-MS resolution, peptide length and charge state. These findings are supported by simulations of Orbitrap transients, and highlight that caution should be exercised in deriving centroid mass values from FT transients that do not support baseline separation of the full isotopic composition.

  1. Robust Decentralized Formation Flight Control

    Directory of Open Access Journals (Sweden)

    Zhao Weihua

    2011-01-01

    Full Text Available Motivated by the idea of multiplexed model predictive control (MMPC, this paper introduces a new framework for unmanned aerial vehicles (UAVs formation flight and coordination. Formulated using MMPC approach, the whole centralized formation flight system is considered as a linear periodic system with control inputs of each UAV subsystem as its periodic inputs. Divided into decentralized subsystems, the whole formation flight system is guaranteed stable if proper terminal cost and terminal constraints are added to each decentralized MPC formulation of the UAV subsystem. The decentralized robust MPC formulation for each UAV subsystem with bounded input disturbances and model uncertainties is also presented. Furthermore, an obstacle avoidance control scheme for any shape and size of obstacles, including the nonapriorily known ones, is integrated under the unified MPC framework. The results from simulations demonstrate that the proposed framework can successfully achieve robust collision-free formation flights.

  2. Pulsed hydrogen/deuterium exchange mass spectrometry for time-resolved membrane protein folding studies.

    Science.gov (United States)

    Khanal, Anil; Pan, Yan; Brown, Leonid S; Konermann, Lars

    2012-12-01

    Kinetic folding experiments by pulsed hydrogen/deuterium exchange (HDX) mass spectrometry (MS) are a well-established tool for water-soluble proteins. To the best of our knowledge, the current study is the first that applies this approach to an integral membrane protein. The native state of bacteriorhodopsin (BR) comprises seven transmembrane helices and a covalently bound retinal cofactor. BR exposure to sodium dodecyl sulfate (SDS) induces partial unfolding and retinal loss. We employ a custom-built three-stage mixing device for pulsed-HDX/MS investigations of BR refolding. The reaction is triggered by mixing SDS-denatured protein with bicelles. After a variable folding time (10 ms to 24 h), the protein is exposed to excess D(2) O buffer under rapid exchange conditions. The HDX pulse is terminated by acid quenching after 24 ms. Subsequent off-line analysis is performed by size exclusion chromatography and electrospray MS. These measurements yield the number of protected backbone N-H sites as a function of folding time, reflecting the recovery of secondary structure. Our results indicate that much of the BR secondary structure is formed quite late during the reaction, on a time scale of 10 s and beyond. It is hoped that in the future it will be possible to extend the pulsed-HDX/MS approach employed here to membrane proteins other than BR. Copyright © 2012 John Wiley & Sons, Ltd.

  3. Protein hydrogen exchange measured at single-residue resolution by electron transfer dissociation mass spectrometry

    DEFF Research Database (Denmark)

    Rand, Kasper D; Zehl, Martin; Jensen, Ole Nørregaard

    2009-01-01

    Because of unparalleled sensitivity and tolerance to protein size, mass spectrometry (MS) has become a popular method for measuring the solution hydrogen (1H/2H) exchange (HX) of biologically relevant protein states. While incorporated deuterium can be localized to different regions by pepsin...... proteolysis of the labeled protein, the assignment of deuteriums to individual residues is typically not obtained, thereby limiting a detailed understanding of HX and the dynamics of protein structure. Here we use gas-phase fragmentation of peptic peptides by electron transfer dissociation (ETD) to measure...... the HX of individual amide linkages in the amyloidogenic protein beta2-microglobulin. A comparison of the deuterium levels of 60 individual backbone amides of beta2-microglobulin measured by HX-ETD-MS analysis to the corresponding values measured by NMR spectroscopy shows an excellent correlation...

  4. Hydrogen deuterium exchange mass spectrometry in biopharmaceutical discovery and development - A review.

    Science.gov (United States)

    Deng, Bin; Lento, Cristina; Wilson, Derek J

    2016-10-12

    Protein therapeutics have emerged as a major class of biopharmaceuticals over the past several decades, a trend that has motivated the advancement of bioanalytical technologies for protein therapeutic characterization. Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a powerful and sensitive technique that can probe the higher order structure of proteins and has been used in the assessment and development of monoclonal antibodies (mAbs), antibody-drug conjugates (ADCs) and biosimilar antibodies. It has also been used to quantify protein-ligand, protein-receptor and other protein-protein interactions involved in signaling pathways. In manufacturing and development, HDX-MS can validate storage formulations and manufacturing processes for various biotherapeutics. Currently, HDX-MS is being refined to provide additional coverage, sensitivity and structural specificity and implemented on the millisecond timescale to reveal residual structure and dynamics in disordered domains and intrinsically disordered proteins. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Dynamics of urokinase receptor interaction with Peptide antagonists studied by amide hydrogen exchange and mass spectrometry

    DEFF Research Database (Denmark)

    Jørgensen, Thomas J D; Gårdsvoll, Henrik; Danø, Keld

    2004-01-01

    Using amide hydrogen exchange combined with electrospray ionization mass spectrometry, we have in this study determined the number of amide hydrogens on several peptides that become solvent-inaccessible as a result of their high-affinity interaction with the urokinase-type plasminogen activator...... receptor (uPAR). These experiments reveal that at least six out of eight amide hydrogens in a synthetic nine-mer peptide antagonist (AE105) become sequestered upon engagement in uPAR binding. Various uPAR mutants with decreased affinity for this peptide antagonist gave similar results, thereby indicating...... that deletion of the favorable interactions involving the side chains of these residues in uPAR does not affect the number of hydrogen bonds established by the main chain of the peptide ligand. The isolated growth factor-like domain (GFD) of the cognate serine protease ligand for uPAR showed 11 protected amide...

  6. Subzero Celsius separations in three-zone temperature controlled hydrogen deuterium exchange mass spectrometry.

    Science.gov (United States)

    Wales, Thomas E; Fadgen, Keith E; Eggertson, Michael J; Engen, John R

    2017-11-10

    Hydrogen deuterium exchange mass spectrometry (HDX MS) reports on the conformational landscape of proteins by monitoring the exchange between backbone amide hydrogen atoms and deuterium in the solvent. To maintain the label for analysis, quench conditions of low temperature and pH are required during the chromatography step performed after protease digestion but before mass spectrometry. Separation at 0°C is often chosen as this is the temperature where the most deuterium can be recovered without freezing of the typical water and acetonitrile mobile phases. Several recent reports of separations at subzero Celsius emphasize the promise for retaining more deuterium and using a much longer chromatographic gradient or direct infusion time. Here we present the construction and validation of a modified Waters nanoACQUITY HDX manager with a third temperature-controlled zone for peptide separations at subzero temperatures. A new Peltier-cooled door replaces the door of a traditional main cooling chamber and the separations and trapping column are routed through the door housing. To prevent freezing, 35% methanol is introduced post online digestion. No new pumps are required and online digestion is performed as in the past. Subzero separations, using conventional HPLC column geometry of 3μ m particles in a 1×50mm column, did not result in major changes to chromatographic efficiency when lowering the temperature from 0 to -20°C. There were significant increases in deuterium recovery for both model peptides and biologically relevant protein systems. Given the higher levels of deuterium recovery, expanded gradient programs can be used to allow for higher chromatographic peak capacity and therefore the analysis of larger and more complex proteins and systems. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Wage Dispersion and Decentralization of Wage Bargaining

    DEFF Research Database (Denmark)

    Dahl, Christian Møller; le Maire, Christian Daniel; Munch, Jakob R.

    2013-01-01

    This article studies how decentralization of wage bargaining from sector to firm level influences wage levels and wage dispersion. We use detailed panel data covering a period of decentralization in the Danish labor market. The decentralization process provides variation in the individual worker...

  8. Ubiquitous consultation tool for decentral knowledge workers

    OpenAIRE

    Nazari Shirehjini, A.A.; Rühl, C.; Noll, S.

    2003-01-01

    The special issue of this initial study is to examine the current work situation of consulting companies, and to elaborate a concept for supporting decentralized working consultants. The concept addresses significant challenges of decentralized work processes by deploying the Peer-to-Peer methodology to decentralized expert and Knowledge Management, cooperation, and enterprise resource planning.

  9. Denaturation Kinetics of Whey Protein Isolate Solutions and Fouling Mass Distribution in a Plate Heat Exchanger

    Directory of Open Access Journals (Sweden)

    Marwa Khaldi

    2015-01-01

    Full Text Available Few investigations have attempted to connect the mechanism of dairy fouling to the chemical reaction of denaturation (unfolding and aggregation occurring in the bulk. The objective of this study is to contribute to this aspect in order to propose innovative controls to limit fouling deposit formation. Experimental investigations have been carried out to observe the relationship between the deposit mass distribution generated in plate heat exchangers (PHE by a whey protein isolate (WPI mainly composed of β-lactoglobulin (β-Lg and the ratio between the unfolding and aggregation rate constants. Experiments using a PHE were carried out at a pilot scale to identify the deposit distribution of a model fouling solution with different calcium contents. In parallel, laboratory experiments were performed to determine the unfolding/aggregation rate constants. Data analysis showed that (i β-Lg denaturation is highly dependent on the calcium content, (ii for each fouling solution, irrespective of the imposed temperature profile, the deposit mass in each channel and the ratio between the unfolding and aggregation rate constants seem to be well correlated. This study demonstrates that both the knowledge of the thermal profile and the β-Lg denaturation rate constants are required in order to predict accurately the deposit distribution along the PHE.

  10. Decentralized control: Status and outlook

    Czech Academy of Sciences Publication Activity Database

    Bakule, Lubomír

    2014-01-01

    Roč. 38, č. 1 (2014), s. 71-80 ISSN 1367-5788 R&D Projects: GA ČR GA13-02149S Institutional support: RVO:67985556 Keywords : decentralized control * networked control systems * event-triggered approach Subject RIV: BC - Control Systems Theory Impact factor: 2.518, year: 2014

  11. Decentralized control with input saturation

    NARCIS (Netherlands)

    Saberi, Ali; Stoorvogel, Antonie Arij; Sannuti, Peddapullaiah

    In decentralized control it is known that the system can be stabilized only if the so-called fixed modes are all stable. If we have input constraints then (semi-)global stability requires all poles to be in the closed left half plane. This paper establishes that these two requirements are necessary

  12. Peer Matcher : Decentralized Partnership Formation

    NARCIS (Netherlands)

    Bozdog, Nicolae Vladimir; Voulgaris, Spyros; Bal, Henri; van Halteren, Aart

    2015-01-01

    This paper presents Peer Matcher, a fully decentralized algorithm solving the k-clique matching problem. The aim of k-clique matching is to cluster a set of nodes having pair wise weights into k-size groups of maximal total weight. Since solving the problem requires exponential time, Peer Matcher

  13. Tracking lysozyme unfolding during salt-induced precipitation with hydrogen exchange and mass spectrometry.

    Science.gov (United States)

    Tobler, S A; Sherman, N E; Fernandez, E J

    We utilized electrospray ionization mass spectrometry (ESI-MS) and hydrogen-deuterium exchange (HX) to detect unfolding of hen egg white lysozyme during salt-induced precipitation. Deuterated lysozyme was dissolved in protonated buffer at pH 2.16 and precipitated with ammonium sulfate, sodium chloride, and potassium thiocyanate. ESI-MS was used to detect mass differences in lysozyme due to the loss of deuterons for solvent protons, providing insight on the conformational history of the protein during the labeling experiment. Precipitation with ammonium sulfate and sodium chloride did not unfold lysozyme, consistent with the known stabilizing effects of kosmotropic salts. Potassium thiocyanate, an aggressive chaotrope, was an effective precipitant at 0.2 M, but also disrupted lysozyme structure and caused the formation of precipitate fractions that did not readily redissolve into aqueous solution without the use of a chemical denaturant. Precipitation with 1.0 M thiocyanate resulted in faster rates of unfolding and larger amounts of the insoluble precipitate. The unfolding kinetics were biphasic, exhibiting a slow phase after a few hours that presumably reflected a smaller propensity for lysozyme to unfold in the precipitated state. Bimodal mass distributions in the ESI-MS spectra for the thiocyanate precipitates indicate two states for lysozyme in this system, a native and a molten globule-like partially unfolded state. ESI-MS analysis of the insoluble precipitates indicated that they consisted primarily of protein molecules that had unfolded. Investigation of the HX behavior of lysozyme in a KSCN solution at low protein concentrations confirmed the destabilizing effect of the salt on the protein structure, even when there was almost no solid phase present. The HX/ESI-MS results provide insight into the mechanism combining precipitation and denaturation for such a system, both in terms of obtaining quantitative kinetic and stability information and the

  14. Mass-independent isotopic fractionation of tin in a chemical exchange reaction

    Science.gov (United States)

    Fujii, T.; Moynier, F.

    2008-12-01

    Tin isotopes were fractionated by the liquid-liquid extraction technique with a crown ether, dicyclohexano-18- crown-6 (DC18C6). Tin dichloride was dissolved in HCl to create solutions, 0.1 mol dm-3 (M) Sn(II) in various [HCl]. The organic phase was 0.1 M DC18C6 in 1,2-dichloroethane. A 5 cm3 aqueous solution and a 5 cm3 organic solution were mixed in a glass vial with a stirrer bar, and the glass vial was sealed. The two phases were stirred by a magnetic stirrer, and after the equilibrium, the two phases were separated by centrifugation. An aliquot of the upper aqueous solution was taken for analysis. These procedures were carried out at room temperature. The Sn concentration in the equilibrated aqueous phase was analyzed by ICP-AES. In order to avoid any matrix effect, possible trace of organic materials from DC18C6 was separated from the Sn on a micro-column anion-exchange resin in HBr/HNO3. An aliquot of the aqueous phase was once dried by heating and then dissolved into a HBr solution. Sn was strongly adsorbed while organic materials went through. Finally Sn was collected in a HNO3 solution. A solution containing 300 ppb of Sn in 0.05 M HNO3 was prepared for isotopic analysis. Isotopic ratios of Sn in all samples were analyzed with the MC-ICP-MS Nu plasma 500 HR at ENS Lyon coupled with a desolvating nebulizer Nu DSN-100 (with a typical precision of <100 ppm). The instrumental mass bias was controlled by bracketing each of the samples with standards. Because 120Sn is the most abundant isotopes, the isotope ratios have been measured as mSn/120Sn (m : 116, 117, 118, 119, 122, and 124). 114Sn and 115Sn were not analyzed due to their low abundances. Possible interferences with Cd on the masses 116, with Te on the mass 120, 122 and 124 were verified to be under the detection of the ICP-MS. Odd atomic mass isotopes (117Sn and 119Sn) showed deficits of enrichment comparing to the even atomic mass isotopes (116Sn, 118Sn, 122Sn, and 124Sn). This odd-even staggering

  15. Fusion-supported decentralized nuclear energy system

    International Nuclear Information System (INIS)

    Jassby, D.L.

    1979-04-01

    A decentralized nuclear energy system is proposed comprising mass-produced pressurized water reactors in the size range 10 to 300 MW (thermal), to be used for the production of process heat, space heat, and electricity in applications where petroleum and natural gas are presently used. Special attention is given to maximizing the refueling interval with no interim batch shuffling in order to minimize fuel transport, reactor downtime, and opportunity for fissile diversion. These objectives demand a substantial fissile enrichment (7 to 15%). The preferred fissile fuel is U-233, which offers an order of magnitude savings in ore requirements (compared with U-235 fuel), and whose higher conversion ratio in thermal reactors serves to extend the period of useful reactivity and relieve demand on the fissile breeding plants (compared with Pu-239 fuel). Application of the neutral-beam-driven tokamak fusion-neutron source to a U-233 breeding pilot plant is examined. This scheme can be extended in part to a decentralized fusion energy system, wherein remotely located large fusion reactors supply excess tritium to a distributed system of relatively small nonbreeding D-T reactors

  16. Dissection of conformational conversion events during prion amyloid fibril formation using hydrogen exchange and mass spectrometry.

    Science.gov (United States)

    Singh, Jogender; Udgaonkar, Jayant B

    2013-09-23

    A molecular understanding of prion diseases requires an understanding of the mechanism of amyloid fibril formation by the prion protein. In particular, it is necessary to define the sequence of the structural events describing the conformational conversion of monomeric PrP to aggregated PrP. In this study, the sequence of the structural events in the case of amyloid fibril formation by recombinant mouse prion protein at pH7 has been characterized by hydrogen-deuterium exchange and mass spectrometry. The observation that fibrils are substantially more stable to hydrogen-deuterium exchange than is native monomer allows both forms to be quantified during the course of the aggregation reaction. Under the aggregation conditions utilized, native monomeric protein and amyloid fibrils are the only forms of the protein detectable during the course of the fibril formation reaction, suggesting that monomer directly adds on to the fibril template. Conformational conversion is shown to occur in two steps after the binding of monomer to fibril, with helix 1 unfolding only after helices 2 and 3 transform into β-sheet. Local stability in the β-sheet core region (residues ~159-225) of the fibrils is shown to be sequence dependent in that it varies along the length of the core, and local stability in protein molecules that are ordered in the structurally heterogeneous sequence segment 109-132 is shown to be similar to that in the core. This new understanding of the structural events during prion protein aggregation has important bearing on our comprehension of the molecular basis of prion pathogenesis. © 2013 Elsevier Ltd. All rights reserved.

  17. Automated Hydrogen/Deuterium Exchange Electron Transfer Dissociation High Resolution Mass Spectrometry Measured at Single-Amide Resolution

    OpenAIRE

    Landgraf, Rachelle R.; Chalmers, Michael J.; Griffin, Patrick R.

    2011-01-01

    Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a well established method for the measurement of solution-phase deuterium incorporation into proteins, which can provide insight into protein conformational mobility. However, most HDX measurements are constrained to regions of the protein where pepsin proteolysis allows detection at peptide resolution. Recently, single-amide resolution deuterium incorporation has been achieved by limiting gas-phase scrambling in the mass spectrometer....

  18. The Two Edge Knife of Decentralization

    Directory of Open Access Journals (Sweden)

    Ahmad Khoirul Umam

    2011-07-01

    Full Text Available A centralistic government model has become a trend in a number of developing countries, in which the ideosycretic aspect becomes pivotal key in the policy making. The situation constitutes authoritarianism, cronyism, and corruption. To break the impasse, the decentralized system is proposed to make people closer to the public policy making. Decentralization is also convinced to be the solution to create a good governance. But a number of facts in the developing countries demonstrates that decentralization indeed has ignite emerges backfires such as decentralized corruption, parochialism, horizontal conflict, local political instability and others. This article elaborates the theoretical framework on decentralization's ouput as the a double-edge knife. In a simple words, the concept of decentralization does not have a permanent relationship with the creation of good governance and development. Without substantive democracy, decentralization is indeed potential to be a destructive political instrument threating the state's future.

  19. Decentralized neural control application to robotics

    CERN Document Server

    Garcia-Hernandez, Ramon; Sanchez, Edgar N; Alanis, Alma y; Ruz-Hernandez, Jose A

    2017-01-01

    This book provides a decentralized approach for the identification and control of robotics systems. It also presents recent research in decentralized neural control and includes applications to robotics. Decentralized control is free from difficulties due to complexity in design, debugging, data gathering and storage requirements, making it preferable for interconnected systems. Furthermore, as opposed to the centralized approach, it can be implemented with parallel processors. This approach deals with four decentralized control schemes, which are able to identify the robot dynamics. The training of each neural network is performed on-line using an extended Kalman filter (EKF). The first indirect decentralized control scheme applies the discrete-time block control approach, to formulate a nonlinear sliding manifold. The second direct decentralized neural control scheme is based on the backstepping technique, approximated by a high order neural network. The third control scheme applies a decentralized neural i...

  20. Investigating the Interaction between the Neonatal Fc Receptor and Monoclonal Antibody Variants by Hydrogen/Deuterium Exchange Mass Spectrometry

    DEFF Research Database (Denmark)

    Jensen, Pernille Foged; Larraillet, Vincent; Schlothauer, Tilman

    2015-01-01

    in understanding and modulating the IgG-FcRn interaction to optimize antibody pharmacokinetics and ultimately improve efficacy and safety. Here we studied the interaction between a full-length human IgG1 and human FcRn via hydrogen/deuterium exchange mass spectrometry and targeted electron transfer dissociation......-type glycosylated IgG. Our results provide new molecular insight into the IgG-FcRn interaction and illustrate the capability of hydrogen/deuterium exchange mass spectrometry to advance structural proteomics by providing detailed information on the conformation and dynamics of large protein complexes in solution....... to map sites perturbed by binding on both partners of the IgG-FcRn complex. Several regions in the antibody Fc region and the FcRn were protected from exchange upon complex formation, in good agreement with previous crystallographic studies of FcRn in complex with the Fc fragment. Interestingly, we found...

  1. Nepenthesin from monkey cups for hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Rey, Martial; Yang, Menglin; Burns, Kyle M; Yu, Yaping; Lees-Miller, Susan P; Schriemer, David C

    2013-02-01

    Studies of protein dynamics, structure and interactions using hydrogen/deuterium exchange mass spectrometry (HDX-MS) have sharply increased over the past 5-10 years. The predominant technology requires fast digestion at pH 2-3 to retain deuterium label. Pepsin is used almost exclusively, but it provides relatively low efficiency under the constraints of the experiment, and a selectivity profile that renders poor coverage of intrinsically disordered regions. In this study we present nepenthesin-containing secretions of the pitcher plant Nepenthes, commonly called monkey cups, for use in HDX-MS. We show that nepenthesin is at least 1400-fold more efficient than pepsin under HDX-competent conditions, with a selectivity profile that mimics pepsin in part, but also includes efficient cleavage C-terminal to "forbidden" residues K, R, H, and P. High efficiency permits a solution-based analysis with no detectable autolysis, avoiding the complication of immobilized enzyme reactors. Relaxed selectivity promotes high coverage of disordered regions and the ability to "tune" the mass map for regions of interest. Nepenthesin-enriched secretions were applied to an analysis of protein complexes in the nonhomologous end-joining DNA repair pathway. The analysis of XRCC4 binding to the BRCT domains of Ligase IV points to secondary interactions between the disordered C-terminal tail of XRCC4 and remote regions of the BRCT domains, which could only be identified with a nepenthesin-based workflow. HDX data suggest that stalk-binding to XRCC4 primes a BRCT conformation in these remote regions to support tail interaction, an event which may be phosphoregulated. We conclude that nepenthesin is an effective alternative to pepsin for all HDX-MS applications, and especially for the analysis of structural transitions among intrinsically disordered proteins and their binding partners.

  2. Boundary layers at a dynamic interface: air-sea exchange of heat and mass

    Science.gov (United States)

    Szeri, Andrew

    2017-11-01

    Exchange of mass or heat across a turbulent liquid-gas interface is a problem of critical interest, especially in air-sea transfer of natural and man-made gases involved in climate change. The goal in this research area is to determine the gas flux from air to sea or vice versa. For sparingly soluble non-reactive gases, this is controlled by liquid phase turbulent velocity fluctuations that act on the thin species concentration boundary layer on the liquid side of the interface. If the fluctuations in surface-normal velocity and gas concentration differences are known, then it is possible to determine the turbulent contribution to the gas flux. However, there is no suitable fundamental direct approach in the general case where neither of these quantities can be easily measured. A new approach is presented to deduce key aspects about the near-surface turbulent motions from remote measurements, which allows one to determine the gas transfer velocity, or gas flux per unit area if overall concentration differences are known. The approach is illustrated with conceptual examples.

  3. Probing Conformational Dynamics of Tau Protein by Hydrogen/Deuterium Exchange Mass Spectrometry

    Science.gov (United States)

    Huang, Richard Y.-C.; Iacob, Roxana E.; Sankaranarayanan, Sethu; Yang, Ling; Ahlijanian, Michael; Tao, Li; Tymiak, Adrienne A.; Chen, Guodong

    2018-01-01

    Fibrillization of the microtubule-associated protein tau has been recognized as one of the signature pathologies of the nervous system in Alzheimer's disease, progressive supranuclear palsy, and other tauopathies. The conformational transition of tau in the fibrillization process, tau monomer to soluble aggregates to fibrils in particular, remains unclear. Here we report on the use of hydrogen/deuterium exchange mass spectrometry (HDX-MS) in combination with other biochemical approaches, including Thioflavin S fluorescence measurements, enzyme-linked immunosorbent assay (ELISA), and Western blotting to understand the heparin-induced tau's fibrillization. HDX-MS studies including anti-tau antibody epitope mapping experiments provided molecular level details of the full-length tau's conformational dynamics and its regional solvent accessibility upon soluble aggregates formation. The results demonstrate that R3 region in the full-length tau's microtubule binding repeat region (MTBR) is stabilized in the aggregation process, leaving both N and C terminal regions to be solvent exposed in the soluble aggregates and fibrils. The findings also illustrate the practical utility of orthogonal analytical methodologies for the characterization of protein higher order structure. [Figure not available: see fulltext.

  4. Conformations of JNK3α splice variants analyzed by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Park, Ji Young; Yun, Youngjoo; Chung, Ka Young

    2017-03-01

    c-Jun N-terminal kinases (JNKs) are members of the mitogen-activated protein kinase (MAPK) family that regulate apoptosis, inflammation, cytokine production, and metabolism. MAPKs undergo various splicing within their kinase domains. Unlike other MAPKs, JNKs have alternative splicing at the C-terminus, resulting in long and short variants. Functional or conformational effects due to the elongated C-terminal tail in the long splice variants have not been investigated nor has the conformation of the C-terminal tail been analyzed. Here, we analyzed the conformation of the elongated C-terminal tail and investigated conformational differences between long and short splice variants of JNKs using JNK3α2 and JNK3α1 as models. We adopted hydrogen/deuterium exchange mass spectrometry (HDX-MS) to analyze the conformation. HDX-MS revealed that the C-terminal tail is mostly intrinsically disordered, and that the conformation of the kinase domain of JNK3α2 is more dynamic than that of JNK3α1. The different conformation dynamics between long and short splice variants of JNK3α might affect the cellular functions of JNK3. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Online deuterium hydrogen exchange and protein digestion coupled with ion mobility spectrometry and tandem mass spectrometry.

    Science.gov (United States)

    Donohoe, Gregory C; Arndt, James R; Valentine, Stephen J

    2015-05-19

    Online deuterium hydrogen exchange (DHX) and pepsin digestion (PD) is demonstrated using drift tube ion mobility spectrometry (DTIMS) coupled with linear ion trap (LTQ) mass spectrometry (MS) with electron transfer dissociation (ETD) capabilities. DHX of deuterated ubiquitin, followed by subsequent quenching and digestion, is performed within ∼60 s, yielding 100% peptide sequence coverage. The high reproducibility of the IMS separation allows spectral feature matching between two-dimensional IMS-MS datasets (undeuterated and deuterated) without the need for dataset alignment. Extracted ion drift time distributions (XIDTDs) of deuterated peptic peptides are mobility-matched to corresponding XIDTDs of undeuterated peptic peptides that were identified using collision-induced dissociation (CID). Matching XIDTDs allows a straightforward identification and deuterium retention evaluation for labeled peptides. Aside from the mobility separation, the ion trapping capabilities of the LTQ, combined with ETD, are demonstrated to provide single-residue resolution. Deuterium retention for the c- series ions across residues M(1)-L(15) and N(25)-R(42) are in good agreement with the known secondary structural elements within ubiquitin.

  6. Differential hydrogen/deuterium exchange mass spectrometry analysis of protein-ligand interactions.

    Science.gov (United States)

    Chalmers, Michael J; Busby, Scott A; Pascal, Bruce D; West, Graham M; Griffin, Patrick R

    2011-02-01

    Functional regulation of ligand-activated receptors is driven by alterations in the conformational dynamics of the protein upon ligand binding. Differential hydrogen/deuterium exchange (HDX) coupled with mass spectrometry has emerged as a rapid and sensitive approach for characterization of perturbations in conformational dynamics of proteins following ligand binding. While this technique is sensitive to detecting ligand interactions and alterations in receptor dynamics, it also can provide important mechanistic insights into ligand regulation. For example, HDX has been used to determine a novel mechanism of ligand activation of the nuclear receptor peroxisome proliferator activated receptor-γ, perform detailed analyses of binding modes of ligands within the ligand-binding pocket of two estrogen receptor isoforms, providing insight into selectivity, and helped classify different types of estrogen receptor-α ligands by correlating their pharmacology with the way they interact with the receptor based solely on hierarchical clustering of receptor HDX signatures. Beyond small-molecule-receptor interactions, this technique has also been applied to study protein-protein complexes, such as mapping antibody-antigen interactions. In this article, we summarize the current state of the differential HDX approaches and the future outlook. We summarize how HDX analysis of protein-ligand interactions has had an impact on biology and drug discovery.

  7. The structural basis of serpin polymerization studied by hydrogen/deuterium exchange and mass spectrometry.

    Science.gov (United States)

    Tsutsui, Yuko; Kuri, Barbara; Sengupta, Tanusree; Wintrode, Patrick L

    2008-11-07

    The serpinopathies are a group of inherited disorders that share as their molecular basis the misfolding and polymerization of serpins, an important class of protease inhibitors. Depending on the identity of the serpin, conditions arising from polymerization include emphysema, thrombosis, and dementia. The structure of serpin polymers is thus of considerable medical interest. Wild-type alpha(1)-antitrypsin will form polymers upon incubation at moderate temperatures and has been widely used as a model system for studying serpin polymerization. Using hydrogen/deuterium exchange and mass spectrometry, we have obtained molecular level structural information on the alpha(1)-antitrypsin polymer. We found that the flexible reactive center loop becomes strongly protected upon polymerization. We also found significant increases in protection in the center of beta-sheet A and in helix F. These results support a model in which linkage between serpins is achieved through insertion of the reactive center loop of one serpin into beta-sheet A of another. We have also examined the heat-induced conformational changes preceding polymerization. We found that polymerization is preceded by significant destabilization of beta-sheet C. On the basis of our results, we propose a mechanism for polymerization in which beta-strand 1C is displaced from the rest of beta-sheet C through a binary serpin/serpin interaction. Displacement of strand 1C triggers further conformational changes, including the opening of beta-sheet A, and allows for subsequent polymerization.

  8. Differential hydrogen/deuterium exchange mass spectrometry analysis of protein–ligand interactions

    Science.gov (United States)

    Chalmers, Michael J; Busby, Scott A; Pascal, Bruce D; West, Graham M; Griffin, Patrick R

    2011-01-01

    Functional regulation of ligand-activated receptors is driven by alterations in the conformational dynamics of the protein upon ligand binding. Differential hydrogen/deuterium exchange (HDX) coupled with mass spectrometry has emerged as a rapid and sensitive approach for characterization of perturbations in conformational dynamics of proteins following ligand binding. While this technique is sensitive to detecting ligand interactions and alterations in receptor dynamics, it also can provide important mechanistic insights into ligand regulation. For example, HDX has been used to determine a novel mechanism of ligand activation of the nuclear receptor peroxisome proliferator activated receptor-γ, perform detailed analyses of binding modes of ligands within the ligand-binding pocket of two estrogen receptor isoforms, providing insight into selectivity, and helped classify different types of estrogen receptor-α ligands by correlating their pharmacology with the way they interact with the receptor based solely on hierarchical clustering of receptor HDX signatures. Beyond small-molecule–receptor interactions, this technique has also been applied to study protein–protein complexes, such as mapping antibody–antigen interactions. In this article, we summarize the current state of the differential HDX approaches and the future outlook. We summarize how HDX analysis of protein–ligand interactions has had an impact on biology and drug discovery. PMID:21329427

  9. Mathematical model of heat-mass exchange processes in a flat solar collector SUN 1

    Directory of Open Access Journals (Sweden)

    Tunik Aleksandr Aleksandrovich

    2016-01-01

    Full Text Available In a flat solar collector SUN 1 The active development of environmental friendly energy sources alternative to HPPs is currently of great importance in the world. Such alternative energy sources are: water, ground, sun, wind, biofuel, etc. If we have a look at the atlas of solar energy resources on the territory of Russia, we can make a conclusion, that in many regions of our country solar activity level allows using solar collector. Though the analysis of different models of solar collector showed, that most of them are ineffective in the regions with cold climate, though the solar activity of these regions is of a great level. In this regard, a mathematical model of heat-mass exchange processes in flat solar collectors is introduced in this article. The model was a basis for the development of a new solar collector, named SUN 1, which has an original heating tubes form. This form allows heat transfer medium to be under the influence of solar energy for a longer time and consequently to warm to a higher temperature, increasing the warming rapidity.

  10. Decentralized stabilization of semi-active vibrating structures

    Science.gov (United States)

    Pisarski, Dominik

    2018-02-01

    A novel method of decentralized structural vibration control is presented. The control is assumed to be realized by a semi-active device. The objective is to stabilize a vibrating system with the optimal rates of decrease of the energy. The controller relies on an easily implemented decentralized switched state-feedback control law. It uses a set of communication channels to exchange the state information between the neighboring subcontrollers. The performance of the designed method is validated by means of numerical experiments performed for a double cantilever system equipped with a set of elastomers with controlled viscoelastic properties. In terms of the assumed objectives, the proposed control strategy significantly outperforms the passive damping cases and is competitive with a standard centralized control. The presented methodology can be applied to a class of bilinear control systems concerned with smart structural elements.

  11. QUDeX-MS: hydrogen/deuterium exchange calculation for mass spectra with resolved isotopic fine structure.

    Science.gov (United States)

    Salisbury, Joseph P; Liu, Qian; Agar, Jeffrey N

    2014-12-11

    Hydrogen/deuterium exchange (HDX) coupled to mass spectrometry permits analysis of structure, dynamics, and molecular interactions of proteins. HDX mass spectrometry is confounded by deuterium exchange-associated peaks overlapping with peaks of heavy, natural abundance isotopes, such as carbon-13. Recent studies demonstrated that high-performance mass spectrometers could resolve isotopic fine structure and eliminate this peak overlap, allowing direct detection and quantification of deuterium incorporation. Here, we present a graphical tool that allows for a rapid and automated estimation of deuterium incorporation from a spectrum with isotopic fine structure. Given a peptide sequence (or elemental formula) and charge state, the mass-to-charge ratios of deuterium-associated peaks of the specified ion is determined. Intensities of peaks in an experimental mass spectrum within bins corresponding to these values are used to determine the distribution of deuterium incorporated. A theoretical spectrum can then be calculated based on the estimated distribution of deuterium exchange to confirm interpretation of the spectrum. Deuterium incorporation can also be detected for ion signals without a priori specification of an elemental formula, permitting detection of exchange in complex samples of unidentified material such as natural organic matter. A tool is also incorporated into QUDeX-MS to help in assigning ion signals from peptides arising from enzymatic digestion of proteins. MATLAB-deployable and standalone versions are available for academic use at qudex-ms.sourceforge.net and agarlabs.com . Isotopic fine structure HDX-MS offers the potential to increase sequence coverage of proteins being analyzed through mass accuracy and deconvolution of overlapping ion signals. As previously demonstrated, however, the data analysis workflow for HDX-MS data with resolved isotopic fine structure is distinct. QUDeX-MS we hope will aid in the adoption of isotopic fine structure HDX

  12. Problems in decentralized data acquisition

    International Nuclear Information System (INIS)

    Eder, R.

    1985-04-01

    This paper describes INIS (International Nuclear Information System) which is operated by the International Atomic Energy Agency (IAEA) in collaboration with 73 Member States and 14 international organizations. INIS is a computerized system for collecting, processing and disseminating nuclear information. The collection and scanning of literature, input preparation and the dissemination of output are completely decentralized, the checking and merging of the information data are centralized. This paper shows the structure, management, processing and problem areas of this system. (Author)

  13. Decentralization and REDD+ in Brazil

    Directory of Open Access Journals (Sweden)

    Fabiano Toni

    2011-01-01

    Full Text Available Recent discussions on REDD+ (Reducing Emissions from Deforestation and Forest Degradation, plus conservation, sustainable management of forests and enhancement of forest carbon stocks have raised optimism about reducing carbon emissions and deforestation in tropical countries. If approved under the United Nations Framework Convention on Climate Change (UNFCCC, REDD+ mechanisms may generate a substantial influx of financial resources to developing countries. Some authors argue that this money could reverse the ongoing process of decentralization of forest policies that has spread through a large number of developing countries in the past two decades. Central states will be accountable for REDD+ money, and may be compelled to control and keep a significant share of REDD+ funds. Supporters of decentralization argue that centralized implementation of REDD+ will be ineffective and inefficient. In this paper, I examine the relation between subnational governments and REDD+ in Brazil. Data show that some state governments in the Brazilian Amazon have played a key role in creating protected areas (PAs after 2003, which helped decrease deforestation rates. Governors have different stimuli for creating PAs. Some respond to the needs of their political constituency; others have expectations to boost the forest sector so as to increase fiscal revenues. Governors also have led the discussion on REDD+ in Brazil since 2008. Considering their interests and political power, REDD+ is unlikely to curb decentralization in Brazil.

  14. Slow Histidine H/D Exchange Protocol for Thermodynamic Analysis of Protein Folding and Stability using Mass Spectrometry

    OpenAIRE

    Tran, Duc T.; Banerjee, Sambuddha; Alayash, Abdu I.; Crumbliss, Alvin L.; Fitzgerald, Michael C.

    2012-01-01

    Described here is a mass spectrometry based protocol to study the thermodynamic stability of proteins and protein-ligand complexes using the slow H/D exchange reaction of the imidazole C2 proton in histidine side chains. The protocol, which involves evaluating the denaturant dependence of this slow H/D exchange reaction in proteins, allows the global and/or subglobal unfolding/refolding properties of proteins and protein-ligand complexes to be probed. The protocol is developed using several m...

  15. Deglycosylation induces extensive dynamics changes in α-amylase revealed by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Ji, Chengjie; Wei, Gary

    2013-12-15

    N-Linked glycosylation plays important roles in modulating protein structure and function. The direct impact of the modification on protein conformation is not yet well understood. Here we probed the dynamic changes following Endo H trimming of high mannose glycans in α-amylase by means of amide hydrogen/deuterium exchange mass spectrometry. The results revealed that deglycosylation elicited extensive alterations in backbone dynamics, affecting regions both adjacent to and distal from the glycosylation site. The overall exchange rate is reduced in the glycosylated state, which indicates rigidity enhancement due to the attached carbohydrates. Copyright © 2013 John Wiley & Sons, Ltd.

  16. CO2 and O2 Gas Exchange in an Experimental Model of the Btlss with Plant Wastes and Human Wastes Included in the Mass Exchange

    Science.gov (United States)

    Ushakova, Sofya; Tikhomirov, Alexander A.; Velichko, Vladimir; Tikhomirova, Natalia; Trifonov, Sergey V.

    2016-07-01

    Mass exchange processes in the new experimental model of the biotechnical life support system (BTLSS) constructed at the Institute of Biophysics SB RAS have a higher degree of closure than in the previous BTLSS, and, thus, the technologies employed in the new system are more complex. Therefore, before closing the loops of mass exchange processes for several months, the new model of the BTLSS was run to match the technologies employed to cultivate plants and the methods used to involve inedible plant parts and human wastes into the mass exchange with the CO2 absorption rate and the amount of the resulting O2. The plant compartment included vegetables grown on the soil-like substrate (SLS) (chufa, beet, carrot, radish, and lettuce), plants hydroponically grown on expanded clay aggregate (wheat, soybean, watercress), and plants grown in aquaculture (common glasswort and watercress). Nutrient solutions for hydroponically grown plants were prepared by using products of physicochemical mineralization of human wastes. Growing the plants in aquaculture enabled maintaining NaCl concentration in the irrigation solution for hydroponically grown plants at a level safe for the plants. Inedible plant biomass was added to the SLS. Three cycles of closing the system were run, which lasted 7, 7, and 10 days. The comparison of the amount of CO2 fed into the system over 24 h (simulating human respiration) and the amount of CO2 daily exhaled by a 70-kg middle-aged human showed that between 1% and 4% of the daily emissions of CO2 were assimilated in the system, and about 3% of the average human daily O2 requirement accumulated in the system. Plant productivity was between 4 and 4.7% of the human daily vegetable requirement, or between 3 and 3.5% of the total human daily food requirement. Thus, testing of the BTLSS showed a match between the technologies employed to arrange mass exchange processes. This study was supported by the grant of the Russian Science Foundation (Project No. 14-14-00599).

  17. Trends in research on forestry decentralization policies

    DEFF Research Database (Denmark)

    Lund, Jens Friis; Rutt, Rebecca Leigh; Ribot, Jesse

    2018-01-01

    We identify and describe four strands in the literature on forestry decentralization policies: studies that assess impacts of forestry sector decentralization policies on forests and livelihoods; studies that examine whether forestry decentralization empowers public and democratic local...... institutions; studies focusing on power and the role of elites in forestry decentralization, and; studies that historicize and contextualize forestry decentralization as reflective of broader societal phenomena. We argue that these strands reflect disciplinary differences in values, epistemologies, and methods...... preferences, and that they individually provide only partial representations of forestry decentralization policies. Accordingly, we conclude that a comprehensive understanding of these policies cannot rest solely on any of these strands, but should be informed by all of them....

  18. Structural insights into glucan phosphatase dynamics using amide hydrogen-deuterium exchange mass spectrometry.

    Science.gov (United States)

    Hsu, Simon; Kim, Youngjun; Li, Sheng; Durrant, Eric S; Pace, Rachel M; Woods, Virgil L; Gentry, Matthew S

    2009-10-20

    Laforin and starch excess 4 (SEX4) are founding members of a class of phosphatases that dephosphorylate phosphoglucans. Each protein contains a carbohydrate binding module (CBM) and a dual-specificity phosphatase (DSP) domain. The gene encoding laforin is mutated in a fatal neurodegenerative disease called Lafora disease (LD). In the absence of laforin function, insoluble glucans that are hyperphosphorylated and exhibit sparse branching accumulate. It is hypothesized that these accumulations trigger the neurodegeneration and premature death of LD patients. We recently demonstrated that laforin removes phosphate from phosphoglucans and hypothesized that this function inhibits insoluble glucan accumulation. Loss of SEX4 function in plants yields a similar cellular phenotype; an excess amount of insoluble, hyperphosphorylated glucans accumulates in cells. While multiple groups have shown that these phosphatases dephosphorylate phosphoglucans, there is no structure of a glucan phosphatase and little is known about the mechanism whereby they perform this action. We utilized hydrogen-deuterium exchange mass spectrometry (DXMS) and structural modeling to probe the conformational and structural dynamics of the glucan phosphatase SEX4. We found that the enzyme does not undergo a global conformational change upon glucan binding but instead undergoes minimal rearrangement upon binding. The CBM has improved protection from deuteration when bound to glucans, confirming its role in glucan binding. More interestingly, we identified structural components of the DSP that also have improved protection from deuteration upon glucan addition. To determine the position of these regions, we generated a homology model of the SEX4 DSP. The homology model shows that all of these regions are adjacent to the DSP active site. Therefore, our results suggest that these regions of the DSP participate in the presentation of the phosphoglucan to the active site and provide the first structural

  19. Structural Insights of Glucan Phosphatase Dynamics using Amide Hydrogen/Deuterium Exchange Mass Spectrometry

    Science.gov (United States)

    Hsu, Simon; Kim, Youngjun; Li, Sheng; Durrant, Eric S.; Pace, Rachel M.; Woods, Virgil L.; Gentry, Matthew S.

    2009-01-01

    Laforin and Starch Excess 4 (SEX4) are founding members of a class of phosphatases that dephosphorylate phosphoglucans. Each protein contains a carbohydrate binding module (CBM) and a dual specificity phosphatase (DSP) domain. The gene encoding laforin is mutated in a fatal neurodegenerative disease called Lafora disease (LD). In the absence of laforin function, insoluble glucans accumulate that are hyperphosphorylated and exhibit sparse branching. It is hypothesized that these accumulations trigger the neurodegeneration and premature death of LD patients. We recently demonstrated that laforin removes phosphate from phosphoglucans and hypothesized that this function inhibits insoluble glucan accumulation. Loss of SEX4 function in plants yields a similar cellular phenotype; cells accumulate an excess amount of insoluble, hyperphosphorylated glucans. While multiple groups have shown that these phosphatases dephosphorylate phosphoglucans, there is no structure of a glucan phosphatase and little is known about the mechanism whereby they perform this action. We utilized hydrogen-deuterium exchange mass spectrometry (DXMS) and structural modeling to probe the conformational and structural dynamics of the glucan phosphatase SEX4. We found that the enzyme does not undergo a global conformational change upon glucan binding, but instead undergoes minimal rearrangement upon binding. The CBM undergoes increased protection from deuteration when bound to glucans, confirming its role in glucan binding. More interestingly, we identified structural components of the DSP that also undergo increased protection from deuteration upon glucan addition. To determine the position of these regions, we generated a homology model of the SEX4 DSP. The homology model shows that all of these regions are adjacent the DSP active site. Therefore, our results suggest that these regions of the DSP participate in presenting the phosphoglucan to the active site and provide the first structural analysis

  20. Activation of ClpP protease by ADEP antibiotics: insights from hydrogen exchange mass spectrometry.

    Science.gov (United States)

    Sowole, Modupeola A; Alexopoulos, John A; Cheng, Yi-Qiang; Ortega, Joaquin; Konermann, Lars

    2013-11-15

    The bacterial protease ClpP consists of 14 subunits that assemble into two stacked heptameric rings. The central degradation chamber can be accessed via axial pores. In free ClpP, these pores are obstructed by the N-terminal regions of the seven subunits at either end of the barrel. Acyldepsipeptides (ADEPs) are antibacterial compounds that bind in hydrophobic clefts surrounding the pore region, causing the pores to open up. The ensuing uncontrolled degradation of intracellular proteins is responsible for the antibiotic activity of ADEPs. Recently published X-ray structures yielded conflicting models regarding the conformation adopted by the N-terminal regions in the open state. Here, we use hydrogen/deuterium exchange (HDX) mass spectrometry to obtain complementary insights into the ClpP behavior with and without ADEP1. Ligand binding causes rigidification of the equatorial belt, accompanied by destabilization in the vicinity of the binding clefts. The N-terminal regions undergo rapid deuteration with only minor changes after ADEP1 binding, revealing a lack of stable H-bonding. Our data point to a mechanism where the pore opening mechanism is mediated primarily by changes in the packing of N-terminal nonpolar side chains. We propose that a "hydrophobic plug" causes pore blockage in ligand-free ClpP. ADEP1 binding provides new hydrophobic anchor points that nonpolar N-terminal residues can interact with. In this way, ADEP1 triggers the transition to an open conformation, where nonpolar moieties are clustered around the rim of the pore. This proposed mechanism helps reconcile the conflicting models that had been put forward earlier. © 2013.

  1. Application of Atmospheric Pressure Photoionization H/D-exchange Mass Spectrometry for Speciation of Sulfur-containing Compounds.

    Science.gov (United States)

    Acter, Thamina; Kim, Donghwi; Ahmed, Arif; Ha, Ji-Hyoung; Kim, Sunghwan

    2017-08-01

    Herein we report the observation of atmospheric pressure in-source hydrogen-deuterium exchange (HDX) of thiol group for the first time. The HDX for thiol group was optimized for positive atmospheric pressure photoionization (APPI) mass spectrometry (MS). The optimized HDX-MS was applied for 31 model compounds (thiols, thiophenes, and sulfides) to demonstrate that exchanged peaks were observed only for thiols. The optimized method has been successfully applied to the isolated fractions of sulfur-rich oil samples. The exchange of one and two thiol hydrogens with deuterium was observed in the thiol fraction; no HDX was observed in the other fractions. Thus, the results presented in this study demonstrate that the HDX-MS method using APPI ionization source can be effective for speciation of sulfur compounds. This method has the potential to be used to access corrosion problems caused by thiol-containing compounds. Graphical Abstract ᅟ.

  2. Application of Atmospheric Pressure Photoionization H/D-exchange Mass Spectrometry for Speciation of Sulfur-containing Compounds

    Science.gov (United States)

    Acter, Thamina; Kim, Donghwi; Ahmed, Arif; Ha, Ji-Hyoung; Kim, Sunghwan

    2017-08-01

    Herein we report the observation of atmospheric pressure in-source hydrogen-deuterium exchange (HDX) of thiol group for the first time. The HDX for thiol group was optimized for positive atmospheric pressure photoionization (APPI) mass spectrometry (MS). The optimized HDX-MS was applied for 31 model compounds (thiols, thiophenes, and sulfides) to demonstrate that exchanged peaks were observed only for thiols. The optimized method has been successfully applied to the isolated fractions of sulfur-rich oil samples. The exchange of one and two thiol hydrogens with deuterium was observed in the thiol fraction; no HDX was observed in the other fractions. Thus, the results presented in this study demonstrate that the HDX-MS method using APPI ionization source can be effective for speciation of sulfur compounds. This method has the potential to be used to access corrosion problems caused by thiol-containing compounds.

  3. Mass exchange in an experimental new-generation life support system model based on biological regeneration of environment

    Science.gov (United States)

    Tikhomirov, A. A.; Ushakova, S. A.; Manukovsky, N. S.; Lisovsky, G. M.; Kudenko, Yu. A.; Kovalev, V. S.; Gubanov, V. G.; Barkhatov, Yu. V.; Gribovskaya, I. V.; Zolotukhin, I. G.; Gros, J. B.; Lasseur, Ch.

    An experimental model of a biological life support system was used to evaluate qualitative and quantitative parameters of the internal mass exchange. The photosynthesizing unit included the higher plant component (wheat and radish), and the heterotrophic unit consisted of a soil-like substrate, California warms, mushrooms and microbial microflora. The gas mass exchange involved evolution of oxygen by the photosynthesizing component and its uptake by the heterotroph component along with the formation and maintaining of the SLS structure, growth of mushrooms and California worms, human respiration, and some other processes. Human presence in the system in the form of "virtual human" that at regular intervals took part in the respirative gas exchange during the experiment. Experimental data demonstrated good oxygen/carbon dioxide balance, and the closure of the cycles of these gases was almost complete. The water cycle was nearly 100% closed. The main components in the water mass exchange were transpiration water and the watering solution with mineral elements. Human consumption of the edible plant biomass (grains and roots) was simulated by processing these products by a unique physicochemical method of oxidizing them to inorganic mineral compounds, which were then returned into the system and fully assimilated by the plants. The oxidation was achieved by "wet combustion" of organic biomass, using hydrogen peroxide following a special procedure, which does not require high temperature and pressure. Hydrogen peroxide is produced from the water inside the system. The closure of the cycle was estimated for individual elements and compounds. Stoichiometric proportions are given for the main components included in the experimental model of the system. Approaches to the mathematical modeling of the cycling processes are discussed, using the data of the experimental model. Nitrogen, as a representative of biogmic elements, shows an almost 100% closure of the cycle inside

  4. Analysis of phosphoinositide 3-kinase inhibitors by bottom-up electron-transfer dissociation hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Masson, Glenn R; Maslen, Sarah L; Williams, Roger L

    2017-05-16

    Until recently, one of the major limitations of hydrogen/deuterium exchange mass spectrometry (HDX-MS) was the peptide-level resolution afforded by proteolytic digestion. This limitation can be selectively overcome through the use of electron-transfer dissociation to fragment peptides in a manner that allows the retention of the deuterium signal to produce hydrogen/deuterium exchange tandem mass spectrometry (HDX-MS/MS). Here, we describe the application of HDX-MS/MS to structurally screen inhibitors of the oncogene phosphoinositide 3-kinase catalytic p110α subunit. HDX-MS/MS analysis is able to discern a conserved mechanism of inhibition common to a range of inhibitors. Owing to the relatively minor amounts of protein required, this technique may be utilised in pharmaceutical development for screening potential therapeutics. © 2017 The Author(s).

  5. Decentralized or Centralized Systems for Colleges and Universities?

    Science.gov (United States)

    Heydinger, Richard B.; Norris, Donald M.

    1979-01-01

    Arguments for and against decentralization of data management, analysis, and planning systems are presented. It is suggested that technological advances have encouraged decentralization. Caution in this direction is urged and the development of an articulated decentralization program is proposed. (SF)

  6. Hydrogen/Deuterium Exchange Mass Spectrometry Reveals Mechanistic Details of Activation of Nucleoside Diphosphate Kinases by Oligomerization

    OpenAIRE

    Dautant , Alain; Meyer , Philippe; Georgescauld , Florian

    2017-01-01

    International audience; Most oligomeric proteins become active only after assembly, but why oligomerization is required to support function is not well understood. Here, we address this question using the WT and a destabilized mutant (D93N) of the hexameric nucleoside diphosphate kinase from the pathogen Mycobacterium tuberculosis (Mt-NDPK). The conformational dynamics and oligomeric states of each were analyzed during unfolding/folding by Hydrogen/Deuterium exchange mass spectrometry (HDX-MS...

  7. Protein structure change studied by hydrogen-deuterium exchange, functional labeling, and mass spectrometry

    Science.gov (United States)

    Englander, Joan J.; Del Mar, Charyl; Li, Will; Englander, S. Walter; Kim, Jack S.; Stranz, David D.; Hamuro, Yoshitomo; Woods, Virgil L.

    2003-01-01

    An automated high-throughput, high-resolution deuterium exchange HPLC-MS method (DXMS) was used to extend previous hydrogen exchange studies on the position and energetic role of regulatory structure changes in hemoglobin. The results match earlier highly accurate but much more limited tritium exchange results, extend the analysis to the entire sequence of both hemoglobin subunits, and identify some energetically important changes. Allosterically sensitive amide hydrogens located at near amino acid resolution help to confirm the reality of local unfolding reactions and their use to evaluate resolved structure changes in terms of allosteric free energy. PMID:12773622

  8. Site-Specific Hydrogen Isotope Composition of Propane: Mass spectrometric methods, equilibrium temperature dependence, and kinetics of exchange

    Science.gov (United States)

    Xie, H.; Ponton, C.; Kitchen, N.; Lloyd, M. K.; Lawson, M.; Formolo, M. J.; Eiler, J. M.

    2016-12-01

    Intramolecular isotope ordering can constrain temperatures of synthesis, mechanisms of formation, and/or source substrates of organic compounds. Here we explore site-specific hydrogen isotope variations of propane. Statistical thermodynamic models predict that at equilibrium methylene hydrogen (-CH2-) in propane will be 10's of per mil higher in D/H ratio than methyl hydrogen (-CH3) at geologically relevant temperatures, and that this difference is highly temperature dependent ( 0.5-1 ‰/°C). Chemical-kinetic controls on site-specific D/H in propane could constrain the mechanisms, conditions and extents of propane synthesis or destruction. We have developed a method for measuring the difference in D/H ratio between methylene and methyl hydrogen in propane by gas source mass spectrometry. The data were measured using the Thermo Fisher Double Focusing Sector high resolution mass spectrometer (DFS), and involve comparison of the D/H ratios of molecular ion (C3H8+) and the ethyl fragmental ion (C2H5+). We demonstrate the accuracy and precision of this method through analysis of D-labeled and independently analyzed propanes. In the exchange experiments, propane was heated (100-200 oC) either alone or in the presence of D-enriched water (δD=1,1419 ‰ SMOW), with or without one of several potentially catalytic substrates for hours to weeks. Propane was found to exchange hydrogen with water vigorously at 200 °C in the presence of metal catalysts. In the presence of Ni catalyst, methylene hydrogen exchanges 2.5 times faster than methyl hydrogen. Hydrogen exchange in the presence of Pd catalyst is more effective and can equilibrate hydrogen isotope distribution on propane on the order of 7 days. Isotopic exchange in the presence of natural materials have also been tested, but is only measurable in the methylene group at 200 °C. High catalytic activity of Pd permits attainment of a bracketed, time-invariant equilibrium state that we use to calibrate the site

  9. Comparative charge-exchange mass spectrometric and argon-sensitized radiolytic studies on methanol

    International Nuclear Information System (INIS)

    Jonsson, B.-O.; Lind, J.

    1976-01-01

    It is shown that the formation of CO and CH 4 in the argon-sensitized γ-radiolysis of methanol is dependent on a mechanism of charge exchange with the species Ar + . On the other hand, the ion Ar + 2 formed from Ar* is of minor importance. The formation of CH + 3 from methanol after charge exchange with Ar + is a rapid process, taking about 10 -13 s. (author)

  10. EXPERIMENTAL STUDY OF LOCAL HYDRODYNAMICS AND MASS EXCHANGE PROCESSES OF COOLANT IN FUEL ASSEMBLIES OF PRESSURIZED WATER REACTORS

    Directory of Open Access Journals (Sweden)

    S. M. Dmitriev

    2016-01-01

    Full Text Available The results of experimental studies of local hydrodynamics and mass exchange of coolant flow behind spacer and mixing grids of different structural versions that were developed for fuel assemblies of domestic and foreign nuclear reactors are presented in the article. In order to carry out the study the models of the following fuel assemblies have been fabricated: FA for VVER and VBER, FA-KVADRAT for PWR-reactor and FA for KLT-40C reactor. All the models have been fabricated with a full geometrical similarity with full-scale fuel assemblies. The study was carried out by simulating the flow of coolant in a core by air on an aerodynamic test rig. In order to measure local hydrodynamic characteristics of coolant flow five-channel Pitot probes were used that enable to measure the velocity vector in a point by its three components. The tracerpropane method was used for studying mass transfer processes. Flow hydrodynamics was studied by measuring cross-section velocities of coolant flow and coolant rates according to the model cells. The investigation of mass exchange processes consisted of a study of concentration distribution for tracer in experimental model, in determination of attenuation lengths of mass transfer processes behind mixing grids, in calculating of inter-cellar mass exchange coefficient. The database on coolant flow in fuel assemblies for different types of reactors had been accumulated that formed the basis of the engineering substantiation of reactor cores designs. The recommendations on choice of optimal versions of mixing grids have been taken into consideration by implementers of the JSC “OKBM Afrikantov” when creating commissioned fuel assemblies. The results of the study are used for verification of CFD-codes and CFD programs of detailed cell-by-cell calculation of reactor cores in order to decrease conservatism for substantiation of thermal-mechanical reliability.

  11. Policy Recommendations on Decentralization, Local Power and ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    2010-12-22

    Policy Recommendations on Decentralization, Local Power and Women's Rights. December 22, 2010. Image. The present document comprises a set of policy recommendations that define a global agenda on gender and decentralization. It emerged from the analysis and experiences shared during the Conference and the ...

  12. Decentralized Decision Making Toward Educational Goals.

    Science.gov (United States)

    Monahan, William W.; Johnson, Homer M.

    This monograph provides guidelines to help those school districts considering a more decentralized form of management. The authors discuss the levels at which different types of decisions should be made, describe the changing nature of the educational environment, identify different centralization-decentralization models, and suggest a flexible…

  13. Decentralization in Botswana: the reluctant process | Dipholo ...

    African Journals Online (AJOL)

    Botswana\\'s decentralization process has always been justified in terms of democracy and development. Consequently, the government has always argued that it is fully committed to decentralization in order to promote popular participation as well as facilitating sustainable rural development. Yet the government does not ...

  14. Policy Recommendations on Decentralization, Local Power and ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Policy Recommendations on Decentralization, Local Power and Women's Rights. 22 décembre 2010. Image. The present document comprises a set of policy recommendations that define a global agenda on gender and decentralization. It emerged from the analysis and experiences shared during the Conference and the ...

  15. Ion-exchange chromatography/electrospray mass spectrometry for the identification of organic and inorganic species in topiramate tablets.

    Science.gov (United States)

    Xiang, X; Ko, C Y; Guh, H Y

    1996-11-01

    An ion-exchange chromatograph/electrospray ionization mass spectrometer (IC/ESI-MS) was used successfully to identify organic and inorganic species present in topiramate tablets. An ion suppressor is placed between the column and detectors to replace sodium ions in the mobile phase with hydrogen ions supplied by the suppressor. The ensuing combination of the hydrogen ions with the mobile phase hydroxide ions produces water and thus allows simultaneous ion detection by an ion conductivity detector and a mass spectrometer. Analytes, including lactate, glycolate, chloride, formate, sulfate, and oxalate, were unambiguously identified by matching the mass spectra and retention times with those of the authentic compounds. Due to its capability of detecting positive and negative as well as neutral species, ESI-MS provides valuable information which is not available with ion conductivity detection alone. Though the coupling of ion-exchange chromatography to mass spectrometry has been reported previously, this is the first demonstration of IC/ESI-MS for the identification of unknown species in real samples. Finally, with the use of deuterium/carbon-13 labeling and MS/MS techniques, we have confirmed that oxalic acid (HOOC-COOH) is formed from formic acid (HCOOH) at the electrospray interface in the presence of the electric field. This observation not only confirms the identity of an unknown peak, but it also provides new insight into chemistry that can take place during electrospray ionization.

  16. Decentralized Energy from Waste Systems

    Directory of Open Access Journals (Sweden)

    Blanca Antizar-Ladislao

    2010-01-01

    Full Text Available In the last five years or so, biofuels have been given notable consideration worldwide as an alternative to fossil fuels, due to their potential to reduce greenhouse gas emissions by partial replacement of oil as a transport fuel. The production of biofuels using a sustainable approach, should consider local production of biofuels, obtained from local feedstocks and adapted to the socio-economical and environmental characteristics of the particular region where they are developed. Thus, decentralized energy from waste systems will exploit local biomass to optimize their production and consumption. Waste streams such as agricultural and wood residues, municipal solid waste, vegetable oils, and algae residues can all be integrated in energy from waste systems. An integral optimization of decentralized energy from waste systems should not be based on the optimization of each single process, but the overall optimization of the whole process. This is by obtaining optimal energy and environmental benefits, as well as collateral beneficial co-products such as soil fertilizers which will result in a higher food crop production and carbon dioxide fixation which will abate climate change.

  17. Imidazole C-2 Hydrogen/Deuterium Exchange Reaction at Histidine for Probing Protein Structure and Function with MALDI Mass Spectrometry

    Science.gov (United States)

    Hayashi, Naoka; Kuyama, Hiroki; Nakajima, Chihiro; Kawahara, Kazuki; Miyagi, Masaru; Nishimura, Osamu; Matsuo, Hisayuki; Nakazawa, Takashi

    2015-01-01

    We present a mass spectrometric method for analyzing protein structure and function, based on the imidazole C-2 or histidine Cε1 hydrogen/deuterium (H/D) exchange reaction, which is intrinsically second order with respect to the concentrations of the imidazolium cation and OD− in D2O. The second-order rate constant (k2) of this reaction was calculated from the pH-dependency of the pseudo-first-order rate constant (kφ) obtained from the change of average mass ΔMr (0 ≤ ΔMr exchange rate in terms of log(k2max/k2) representing the deviation of k2 from k2max. In the catalytic site of bovine ribonuclease A, His12 showed much larger change in log(k2max/k2) compared with His119 upon binding with cytidine 3′-monophosphate, as anticipated from the X-ray structures and the possible change in solvent accessibility. However, there is a need of considering the hydrogen bonds of the imidazole group with non-dissociable groups to interpret an extremely slow H/D exchange rate of His48 in partially solvent-exposed situation. PMID:24606199

  18. High-resolution mass spectrometry and hydrogen/deuterium exchange study of mitorubrin azaphilones and nitrogenized analogues.

    Science.gov (United States)

    Svilar, Ljubica; Stankov-Jovanovic, Vesna; Lesage, Denis; Dossmann, Héloïse; Tabet, Jean-Claude

    2012-08-01

    Azaphilones represent numerous groups of wild fungal secondary metabolites that exhibit exceptional tendency to bind to nitrogen atoms in various molecules, especially those containing the amine group. Nitrogenized analogues of mitorubrin azaphilones, natural secondary metabolites of Hypoxylon fragiforme fungus, have been detected in the fungal methanol extract in very low concentrations. Positive electrospray ionization interfaced with high-resolution mass spectrometry was applied for confirmation of the elemental composition of protonated species. Collision-induced dissociation (CID) experiments have been performed, and fragmentation mechanisms have been proposed. Additional information regarding both secondary metabolite analogue families has been reached by application of gas-phase proton/deuterium (H/D) exchanges performed in the collision cell of a triple quadrupole mass spectrometer. An incomplete H/D exchange with one proton less than expected was observed for both protonated mitorubrin azaphilones and their nitrogenized analogues. By means of the density functional theory, an appropriate explanation of this behavior was provided, and it revealed some information concerning gas-phase H/D exchange mechanism and protonation sites. Copyright © 2012 John Wiley & Sons, Ltd.

  19. Rapid Conformational Analysis of Protein Drugs in Formulation by Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS)

    DEFF Research Database (Denmark)

    Esmail Nazari, Zeinab; van de Weert, Marco; Bou-Assaf, George

    2016-01-01

    Hydrogen Deuterium Exchange coupled to Mass Spectrometry (HDX-MS) has become an established method for analysis of protein higher-order structure. Here, we use HDX-MS methodology based on manual Solid-Phase Extraction (SPE) to allow fast and simplified conformational analysis of proteins under...... pharmaceutically relevant formulation conditions. Of significant practical utility, the methodology allows global HDX-MS analyses to be performed without refrigeration or external cooling of the setup. In Mode 1, we used DMSO-containing solvents for SPE, allowing the HDX-MS analysis to be performed at acceptable...... in formulation, using an internal HDX reference peptide (P7I) to control for any sample-to-sample variations in back exchange. Advantages of the methodology include low sample use, optimized excipient removal using multiple solvents, and fast data acquisition. Our results indicate that the SPE-HDX-MS system can...

  20. Mass and Heat Transfer in Ion-Exchange Membranes Applicable to Solid Polymer Fuel Cells

    Energy Technology Data Exchange (ETDEWEB)

    Otteroey, M.

    1996-04-01

    In this doctoral thesis, an improved emf method for determination of transference numbers of two counter ions in ion-exchange membranes is presented. Transference numbers were obtained as a continuous function of the composition. The method avoids problems with diffusion by using a stack of membranes. Water transference coefficients in ion-exchange membranes is discussed and reversible and irreversible water transfer is studied by emf methods. Efforts were made to get data relevant to the solid polymer fuel cell. The results support the findings of other researchers that the reversible water transfer is lower than earlier predicted. A chapter on the conductivity of ion-exchange membranes establishes a method to separate the very thin liquid layers surrounding the membranes in a stack. Using the method it was found that the conductivity is obtained with high accuracy and that the liquid layer in a membrane stack can contribute significantly to the total measured resistance. A four point impedance method was tested to measure the conductivity of membranes under fuel cell conditions. Finally, there is a discussion of reversible heat effects and heat transfer in ion-exchange membranes. 155 refs., 45 figs., 13 tabs.

  1. Stagnant mobile phase mass transfer in chromatographic media: Intraparticle diffusion and exchange kinetics

    NARCIS (Netherlands)

    Tallarek, U.; Vergeldt, F.J.; As, van H.

    1999-01-01

    Pulsed field gradient nuclear magnetic resonance has been successfully applied to a direct and detailed experimental study of topological and dynamic aspects involved in the exchange of small, nonsorbed fluid molecules between the intraparticle pore network and the interparticle void space in

  2. Deuterium Exchange in Ethyl Acetoacetate: An Undergraduate GC-MS [Gas Chromatography-Mass Spectroscopy] Experiment

    Science.gov (United States)

    Heinson, C. D.; Williams, J. M.; Tinnerman, W. N.; Malloy, T. B.

    2005-01-01

    The role of ethanol O-d in nullifying the deuterolysis may be demonstrated by determining that transesterification of methyl acetoacetate of the ethyl ester occurs as well as deuterium exchange of the five acetoacetate hydrogens. The significant acidity of the methylene protons in the acetoacetate group, the efficacy of base catalysis, the role of…

  3. Conformational Analysis of Proteins in Highly Concentrated Solutions by Dialysis-Coupled Hydrogen/Deuterium Exchange Mass Spectrometry

    DEFF Research Database (Denmark)

    Houde, Damian; Esmail Nazari, Zeinab; Bou-Assaf, George M

    2016-01-01

    for these phenomena can be due to short range electrostatic and/or hydrophobic protein-protein interactions. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a useful tool for investigating protein conformation, dynamics, and interactions. However, "traditional" continuous dilution labeling HDX......-MS experiments have limited utility for the direct analysis of solutions with high concentrations of protein. Here, we present a dialysis-based HDX-MS (di-HDX-MS) method as an alternative HDX-MS labeling format, which takes advantage of passive dialysis rather than the classic dilution workflow. We applied...

  4. Characterization of Aggregation Propensity of a Human Fc-Fusion Protein Therapeutic by Hydrogen/Deuterium Exchange Mass Spectrometry

    Science.gov (United States)

    Huang, Richard Y.-C.; Iacob, Roxana E.; Krystek, Stanley R.; Jin, Mi; Wei, Hui; Tao, Li; Das, Tapan K.; Tymiak, Adrienne A.; Engen, John R.; Chen, Guodong

    2017-05-01

    Aggregation of protein therapeutics has long been a concern across different stages of manufacturing processes in the biopharmaceutical industry. It is often indicative of aberrant protein therapeutic higher-order structure. In this study, the aggregation propensity of a human Fc-fusion protein therapeutic was characterized. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) was applied to examine the conformational dynamics of dimers collected from a bioreactor. HDX-MS data combined with spatial aggregation propensity calculations revealed a potential aggregation interface in the Fc domain. This study provides a general strategy for the characterization of the aggregation propensity of Fc-fusion proteins at the molecular level.

  5. Hydrogen/deuterium exchange mass spectrometry for probing higher order structure of protein therapeutics: methodology and applications.

    Science.gov (United States)

    Wei, Hui; Mo, Jingjie; Tao, Li; Russell, Reb J; Tymiak, Adrienne A; Chen, Guodong; Iacob, Roxana E; Engen, John R

    2014-01-01

    The higher order structure of protein therapeutics can be interrogated with hydrogen/deuterium exchange mass spectrometry (HDX-MS). HDX-MS is now a widely used tool in the structural characterization of protein therapeutics. In this review, HDX-MS based workflows designed for protein therapeutic discovery and development processes are presented, focusing on the specific applications of epitope mapping for protein/drug interactions and biopharmaceutical comparability studies. Future trends in the application of HDX-MS in protein therapeutics characterization are also described. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Conformational analysis of Epac activation using amide hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Brock, Melissa; Fan, Fenghui; Mei, Fang C; Li, Sheng; Gessner, Christopher; Woods, Virgil L; Cheng, Xiaodong

    2007-11-02

    Exchange proteins directly activated by cAMP (Epac) play important roles in mediating the effects of cAMP through the activation of downstream small GTPases, Rap. To delineate the mechanism of Epac activation, we probed the conformation and structural dynamics of Epac using amide hydrogen/deuterium exchange and structural modeling. Our studies show that cAMP induces significant conformational changes that lead to a spatial rearrangement of the regulatory components of Epac and allows the exposure of the catalytic core for effector binding without imposing significant conformational change on the catalytic core. Homology modeling and comparative structural analyses of the cAMP binding domains of Epac and cAMP-dependent protein kinase (PKA) lead to a model of Epac activation, in which Epac and PKA activation by cAMP employs the same underlying principle, although the detailed structural and conformational changes associated with Epac and PKA activation are significantly different.

  7. Simulated North Atlantic-Nordic Seas water mass exchanges in an isopycnic coordinate OGCM

    OpenAIRE

    Nilsen, Jan Even Øie; Gao, Yongqi; Drange, Helge; Furevik, Tore; Bentsen, Mats

    2003-01-01

    The variability in the volume exchanges between the North Atlantic and the Nordic Seas during the last 50 years is investigated using a synoptic forced, global version of the Miami Isopycnic Coordinate Ocean Model (MICOM). The simulated volume fluxes agree with the existing observations. The net volume flux across the Faroe-Shetland Channel (FSC) is positively correlated with the net flux through the Denmark Strait (DS; R = 0.74 for 3 years low pass filtering), but negatively correlated with ...

  8. Mass exchange and angular distribution in a dynamical treatment of heavy ion collisions

    International Nuclear Information System (INIS)

    Ngo, C.; Hofmann, H.

    1977-01-01

    One presents a first numerical computation of the absolute value of the double differential cross section as a function of mass asymmetry and detection angle including a dynamical coupling between relative motion and mass asymmetry. One applies it to the 63 Cu+ 197 Au experiment at two different energies. The equation of motion used is a Fokker-Planck equation for distribution function in classical phase space. The coefficients needed are those known from classical model calculations, besides a friction coefficient introduced for the mass asymmetry degree. Encouraging agreement between the calculated and experimental curves is found

  9. Conformational changes in Akt1 activation probed by amide hydrogen/deuterium exchange and nano-electrospray ionization mass spectrometry†

    Science.gov (United States)

    Guo, Mingquan; Huang, Bill X.; Kim, Hee-Yong

    2009-01-01

    Amide hydrogen exchange coupled to nano-electrospray ionization mass spectrometry (nano-ESI-MS) has been used to identify and characterize localized conformational changes of Akt upon activation. Active or inactive Akt was incubated in D2O buffer, digested with pepsin, and analyzed by nano-ESI-MS to determine the deuterium incorporation. The hydrogen/deuterium (H/D) exchange profiles revealed that Akt undergoes considerable conformational changes in the core structures of all three individual domains after activation. In the PH domain, four β-strand (β1, β2 β5 and β6) regions containing membrane-binding residues displayed higher solvent accessibility in the inactive state, suggesting that the PH domain is readily available for the binding to the plasma membrane for activation. In contrast, these β-strands became less exposed or more folded in the active form, which is favored for the dissociation of Akt from the membrane. The beginning α-helix J region and the C-terminal locus (T450-470P) of the regulatory domain showed less folded structures that probably enable substrate entry. Our data also revealed detailed conformational changes of Akt in the kinase domain due to activation, some of which may be attributed to the interaction of the basic residues with phosphorylation sites. Our H/D exchange results indicating the conformational status of Akt at different activation states provided new insight for the regulation of this critical protein involved in cell survival. PMID:19462409

  10. Computing for Decentralized Systems (lecture 2)

    CERN Multimedia

    CERN. Geneva

    2018-01-01

    With the rise of Bitcoin, Ethereum, and other cryptocurrencies it is becoming apparent the paradigm shift towards decentralized computing. Computer engineers will need to understand this shift when developing systems in the coming years. Transferring value over the Internet is just one of the first working use cases of decentralized systems, but it is expected they will be used for a number of different services such as general purpose computing, data storage, or even new forms of governance. Decentralized systems, however, pose a series of challenges that cannot be addressed with traditional approaches in computing. Not having a central authority implies truth must be agreed upon rather than simply trusted and, so, consensus protocols, cryptographic data structures like the blockchain, and incentive models like mining rewards become critical for the correct behavior of decentralized system. This series of lectures will be a fast track to introduce these fundamental concepts through working examples and pra...

  11. Computing for Decentralized Systems (lecture 1)

    CERN Multimedia

    CERN. Geneva

    2018-01-01

    With the rise of Bitcoin, Ethereum, and other cryptocurrencies it is becoming apparent the paradigm shift towards decentralized computing. Computer engineers will need to understand this shift when developing systems in the coming years. Transferring value over the Internet is just one of the first working use cases of decentralized systems, but it is expected they will be used for a number of different services such as general purpose computing, data storage, or even new forms of governance. Decentralized systems, however, pose a series of challenges that cannot be addressed with traditional approaches in computing. Not having a central authority implies truth must be agreed upon rather than simply trusted and, so, consensus protocols, cryptographic data structures like the blockchain, and incentive models like mining rewards become critical for the correct behavior of decentralized system. This series of lectures will be a fast track to introduce these fundamental concepts through working examples and pra...

  12. The effects of fiscal decentralization in Albania

    Directory of Open Access Journals (Sweden)

    Dr.Sc. Blerta Dragusha

    2012-06-01

    Full Text Available “Basically decentralization is a democratic reform which seeks to transfer the political, administrative, financial and planning authority from central to local government. It seeks to develop civic participation, empowerment of local people in decision making process and to promote accountability and reliability: To achieve efficiency and effectiveness in the collection and management of resources and service delivery”1 The interest and curiosity of knowing how our country is doing in this process, still unfinished, served as a motivation forme to treat this topic: fiscal decentralization as a process of giving 'power' to local governments, not only in terms of rights deriving from this process but also on the responsibilities that come with it. Which are the stages before and after decentralization, and how has it affected the process in several key indicators? Is decentralization a good process only, or can any of its effects be seen as an disadvantage?

  13. Inclusion of human mineralized exometabolites and fish wastes as a source of higher plant mineral nutrition in BTLSS mass exchange

    Science.gov (United States)

    Tikhomirova, Natalia; Tikhomirov, Alexander A.; Ushakova, Sofya; Anischenko, Olesya; Trifonov, Sergey V.

    Human exometabolites inclusion into an intrasystem mass exchange will allow increasing of a closure level of a biological-technical life support system (BTLSS). Previously at the IBP SB RAS it was shown that human mineralized exometabolites could be incorporated in the BTLSS mass exchange as a mineral nutrition source for higher plants. However, it is not known how that combined use of human mineralized exometabolites and fish wastes in the capacity of nutrient medium, being a part of the BTLSS consumer wastes, will affect the plant productivity. Several wheat vegetations were grown in an uneven-aged conveyor on a neutral substrate. A mixture of human mineralized exometabolites and fish wastes was used as a nutrient solution in the experiment treatment and human mineralized exometabolites were used in the control. Consequently, a high wheat yield in the experiment treatment practically equal to the control yield was obtained. Thus, mineralized fish wastes can be an additional source of macro-and micronutrients for plants, and use of such wastes for the plant mineral nutrition allows increasing of BTLSS closure level.

  14. Phosphoinositide binding regulates α-actinin CH2 domain structure: Analysis by hydrogen/deuterium exchange mass spectrometry

    Science.gov (United States)

    Full, Stephen J.; Deinzer, Max L.; Ho, P. Shing; Greenwood, Jeffrey A.

    2007-01-01

    α-Actinin is an actin bundling protein that regulates cell adhesion by directly linking actin filaments to integrin adhesion receptors. Phosphatidylinositol (4,5)-diphosphate (PtdIns (4,5)-P2) and phosphatidylinositol (3,4,5)-triphosphate (PtdIns (3,4,5)-P3) bind to the calponin homology 2 domain of α-actinin, regulating its interactions with actin filaments and integrin receptors. In this study, we examine the mechanism by which phosphoinositide binding regulates α-actinin function using mass spectrometry to monitor hydrogen–deuterium (H/D) exchange within the calponin homology 2 domain. The overall level of H/D exchange for the entire protein showed that PtdIns (3,4,5)-P3 binding alters the structure of the calponin homology 2 domain increasing deuterium incorporation, whereas PtdIns (4,5)-P2 induces changes in the structure decreasing deuterium incorporation. Analysis of peptic fragments from the calponin homology 2 domain showed decreased local H/D exchange within the loop region preceding helix F with both phosphoinositides. However, the binding of PtdIns (3,4,5)-P3 also induced increased exchange within helix E. This suggests that the phosphate groups on the fourth and fifth position of the inositol head group of the phosphoinositides constrict the calponin homology 2 domain, thereby altering the orientation of actin binding sequence 3 and decreasing the affinity of α-actinin for filamentous actin. In contrast, the phosphate group on the third position of the inositol head group of PtdIns (3,4,5)-P3 perturbs the calponin homology 2 domain, altering the interaction between the N and C terminus of the full-length α-actinin antiparallel homodimer, thereby disrupting bundling activity and interaction with integrin receptors. PMID:17965186

  15. Phosphoinositide binding regulates alpha-actinin CH2 domain structure: analysis by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Full, Stephen J; Deinzer, Max L; Ho, P Shing; Greenwood, Jeffrey A

    2007-12-01

    alpha-Actinin is an actin bundling protein that regulates cell adhesion by directly linking actin filaments to integrin adhesion receptors. Phosphatidylinositol (4,5)-diphosphate (PtdIns (4,5)-P(2)) and phosphatidylinositol (3,4,5)-triphosphate (PtdIns (3,4,5)-P(3)) bind to the calponin homology 2 domain of alpha-actinin, regulating its interactions with actin filaments and integrin receptors. In this study, we examine the mechanism by which phosphoinositide binding regulates alpha-actinin function using mass spectrometry to monitor hydrogen-deuterium (H/D) exchange within the calponin homology 2 domain. The overall level of H/D exchange for the entire protein showed that PtdIns (3,4,5)-P(3) binding alters the structure of the calponin homology 2 domain increasing deuterium incorporation, whereas PtdIns (4,5)-P(2) induces changes in the structure decreasing deuterium incorporation. Analysis of peptic fragments from the calponin homology 2 domain showed decreased local H/D exchange within the loop region preceding helix F with both phosphoinositides. However, the binding of PtdIns (3,4,5)-P(3) also induced increased exchange within helix E. This suggests that the phosphate groups on the fourth and fifth position of the inositol head group of the phosphoinositides constrict the calponin homology 2 domain, thereby altering the orientation of actin binding sequence 3 and decreasing the affinity of alpha-actinin for filamentous actin. In contrast, the phosphate group on the third position of the inositol head group of PtdIns (3,4,5)-P(3) perturbs the calponin homology 2 domain, altering the interaction between the N and C terminus of the full-length alpha-actinin antiparallel homodimer, thereby disrupting bundling activity and interaction with integrin receptors.

  16. Characterizing the dynamics of α-synuclein oligomers using hydrogen/deuterium exchange monitored by mass spectrometry.

    Science.gov (United States)

    Mysling, Simon; Betzer, Cristine; Jensen, Poul H; Jorgensen, Thomas J D

    2013-12-23

    Soluble oligomers formed by α-synuclein (αSN) are suspected to play a central role in neuronal cell death during Parkinson's disease. While studies have probed the surface structure of these oligomers, little is known about the backbone dynamics of αSN when they form soluble oligomers. Using hydrogen/deuterium exchange monitored by mass spectrometry (HDX-MS), we have analyzed the structural dynamics of soluble αSN oligomers. The analyzed oligomers were metastable, slowly dissociating to monomers over a period of 21 days, after excess monomer had been removed. The C-terminal region of αSN (residues 94-140) underwent isotopic exchange very rapidly, demonstrating a highly dynamic region in the oligomeric state. Three regions (residues 4-17, 39-54, and 70-89) were strongly protected against isotopic exchange in the oligomers, indicating the presence of a stable hydrogen-bonded or solvent-shielded structure. The protected regions were interspersed by two somewhat more dynamic regions (residues 18-38 and 55-70). In the oligomeric state, the isotopic exchange pattern of the region of residues 35-95 of αSN corresponded well with previous nuclear magnetic resonance and electron paramagnetic resonance analyses performed on αSN fibrils and indicated a possible zipperlike maturation mechanism for αSN aggregates. We find the protected N-terminus (residues 4-17) to be of particular interest, as this region has previously been observed to be highly dynamic for both monomeric and fibrillar αSN. This region has mainly been described in relation to membrane binding of αSN, and structuring may be important in relation to disease.

  17. Protein Structure-Function Correlation in Living Human Red Blood Cells Probed by Isotope Exchange-based Mass Spectrometry.

    Science.gov (United States)

    Narayanan, Sreekala; Mitra, Gopa; Muralidharan, Monita; Mathew, Boby; Mandal, Amit K

    2015-12-01

    To gain insight into the underlying mechanisms of various biological events, it is important to study the structure-function correlation of proteins within cells. Structural probes used in spectroscopic tools to investigate protein conformation are similar across all proteins. Therefore, structural studies are restricted to purified proteins in vitro and these findings are extrapolated in cells to correlate their functions in vivo. However, due to cellular complexity, in vivo and in vitro environments are radically different. Here, we show a novel way to monitor the structural transition of human hemoglobin upon oxygen binding in living red blood cells (RBCs), using hydrogen/deuterium exchange-based mass spectrometry (H/DX-MS). Exploiting permeability of D2O across cell membrane, the isotope exchange of polypeptide backbone amide hydrogens of hemoglobin was carried out inside RBCs and monitored using matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). To explore the conformational transition associated with oxygenation of hemoglobin in vivo, the isotope exchange kinetics was simplified using the method of initial rates. RBC might be considered as an in vivo system of pure hemoglobin. Thus, as a proof-of-concept, the observed results were correlated with structural transition of hemoglobin associated with its function established in vitro. This is the first report on structural changes of a protein upon ligand binding in its endogenous environment. The proposed method might be applicable to proteins in their native state, irrespective of location, concentration, and size. The present in-cell approach opens a new avenue to unravel a plethora of biological processes like ligand binding, folding, and post-translational modification of proteins in living cells.

  18. Investigating the Interaction between the Neonatal Fc Receptor and Monoclonal Antibody Variants by Hydrogen/Deuterium Exchange Mass Spectrometry*

    Science.gov (United States)

    Jensen, Pernille Foged; Larraillet, Vincent; Schlothauer, Tilman; Kettenberger, Hubert; Hilger, Maximiliane; Rand, Kasper D.

    2015-01-01

    The recycling of immunoglobulins by the neonatal Fc receptor (FcRn) is of crucial importance in the maintenance of antibody levels in plasma and is responsible for the long half-lives of endogenous and recombinant monoclonal antibodies. From a therapeutic point of view there is great interest in understanding and modulating the IgG–FcRn interaction to optimize antibody pharmacokinetics and ultimately improve efficacy and safety. Here we studied the interaction between a full-length human IgG1 and human FcRn via hydrogen/deuterium exchange mass spectrometry and targeted electron transfer dissociation to map sites perturbed by binding on both partners of the IgG–FcRn complex. Several regions in the antibody Fc region and the FcRn were protected from exchange upon complex formation, in good agreement with previous crystallographic studies of FcRn in complex with the Fc fragment. Interestingly, we found that several regions in the IgG Fab region also showed reduced deuterium uptake. Our findings indicate the presence of hitherto unknown FcRn interaction sites in the Fab region or a possible conformational link between the IgG Fc and Fab regions upon FcRn binding. Further, we investigated the role of IgG glycosylation in the conformational response of the IgG–FcRn interaction. Removal of antibody glycans increased the flexibility of the FcRn binding site in the Fc region. Consequently, FcRn binding did not induce a similar conformational stabilization of deglycosylated IgG as observed for the wild-type glycosylated IgG. Our results provide new molecular insight into the IgG–FcRn interaction and illustrate the capability of hydrogen/deuterium exchange mass spectrometry to advance structural proteomics by providing detailed information on the conformation and dynamics of large protein complexes in solution. PMID:25378534

  19. Investigating the interaction between the neonatal Fc receptor and monoclonal antibody variants by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Jensen, Pernille Foged; Larraillet, Vincent; Schlothauer, Tilman; Kettenberger, Hubert; Hilger, Maximiliane; Rand, Kasper D

    2015-01-01

    The recycling of immunoglobulins by the neonatal Fc receptor (FcRn) is of crucial importance in the maintenance of antibody levels in plasma and is responsible for the long half-lives of endogenous and recombinant monoclonal antibodies. From a therapeutic point of view there is great interest in understanding and modulating the IgG-FcRn interaction to optimize antibody pharmacokinetics and ultimately improve efficacy and safety. Here we studied the interaction between a full-length human IgG(1) and human FcRn via hydrogen/deuterium exchange mass spectrometry and targeted electron transfer dissociation to map sites perturbed by binding on both partners of the IgG-FcRn complex. Several regions in the antibody Fc region and the FcRn were protected from exchange upon complex formation, in good agreement with previous crystallographic studies of FcRn in complex with the Fc fragment. Interestingly, we found that several regions in the IgG Fab region also showed reduced deuterium uptake. Our findings indicate the presence of hitherto unknown FcRn interaction sites in the Fab region or a possible conformational link between the IgG Fc and Fab regions upon FcRn binding. Further, we investigated the role of IgG glycosylation in the conformational response of the IgG-FcRn interaction. Removal of antibody glycans increased the flexibility of the FcRn binding site in the Fc region. Consequently, FcRn binding did not induce a similar conformational stabilization of deglycosylated IgG as observed for the wild-type glycosylated IgG. Our results provide new molecular insight into the IgG-FcRn interaction and illustrate the capability of hydrogen/deuterium exchange mass spectrometry to advance structural proteomics by providing detailed information on the conformation and dynamics of large protein complexes in solution. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. Hydrogen/deuterium exchange mass spectrometry identifies two highly protected regions in recombinant full-length prion protein amyloid fibrils.

    Science.gov (United States)

    Nazabal, Alexis; Hornemann, Simone; Aguzzi, Adriano; Zenobi, Renato

    2009-06-01

    Understanding the structural basis that distinguishes the amyloid form of the prion protein from its monomeric homologue is of crucial importance to elucidate the mechanism of the lethal diseases related to this protein. Recently, an in vitro conversion system was established which reproduces the transition of recombinant prion protein PrP(23-230) from its native alpha-helical rich form into an aggregated amyloid beta-sheet rich form with physicochemical properties reminiscent to those of the disease-related isoform of the prion protein, PrPSc. To study the tertiary and quaternary structural organization within recombinant amyloid fibrils from mouse, mPrP(23-231)betaf; bovine, bPrP(23-230)betaf; and elk, ePrP(23-230)betaf; we utilized hydrogen/deuterium (H/D) exchange analyzed by matrix-assisted laser desorption/ionization (MALDI) and nano-electrospray (nano-ESI) mass spectrometry. No significant differences were found by measuring the deuterium exchange kinetics of the aggregated fibrillar forms for mPrP(23-231)betaf, bPrP(23-230)betaf and ePrP(23-230)betaf, indicating a similar overall structural organization of the fibrils from all three species. Next, we characterized the solvent accessibility for the soluble and fibrillar forms of the mouse prion protein by hydrogen exchange, pepsin proteolysis and nano-ESI ion trap mass spectrometry analysis. In its amyloid form, two highly protected regions of mPrP(23-231) comprising residues [24-98] and [182-212] were identified. The residues between the two highly protected stretches were found to be more solvent exposed, but less than in the soluble protein, and might therefore rather form part of a fibrillar interface. Copyright 2009 John Wiley & Sons, Ltd.

  1. Decentralization and the local development state

    DEFF Research Database (Denmark)

    Emmenegger, Rony Hugo

    2016-01-01

    between the 2005 and 2010 elections. Based on ethnographic field research, the empirical case presented discloses that decentralization and state-led development serve the expansion of state power into rural areas, but that state authority is simultaneously constituted and undermined in the course...... of this process. On that basis, this article aims to contribute to an inherently political understanding of decentralization, development and their entanglement in local and national politics in rural African societies....

  2. Decentralized flight trajectory planning of multiple aircraft

    OpenAIRE

    Yokoyama, Nobuhiro; 横山 信宏

    2008-01-01

    Conventional decentralized algorithms for optimal trajectory planning tend to require prohibitive computational time as the number of aircraft increases. To overcome this drawback, this paper proposes a novel decentralized trajectory planning algorithm adopting a constraints decoupling approach for parallel optimization. The constraints decoupling approach is formulated as the path constraints of the real-time trajectory optimization problem based on nonlinear programming. Due to the parallel...

  3. Interregional Inequality, Decentralization, and Corruption in Indonesia

    OpenAIRE

    Sukadana, I Wayan

    2009-01-01

    Decentralization has been implemented for eight years. However, corruption has not been lessened yet within the decentralization era. Bad regulations and investment climate are suspected to be a cause of the situations. Good regulation and investment climate appear if the interjurisdictional competitions exist. The incentive for competition is the equality of economic endowment among regions, or each region has their own tradeable ”technology” that comparable to others. Inequality in ”...

  4. Decentralized electricity production. v. 1 and 2

    International Nuclear Information System (INIS)

    1991-01-01

    The first part of the symposium is concerned with market analysis, case studies and prospectives for the decentralized production of electricity in France: cogeneration, heat networks, municipal waste incineration, etc. Financing systems and microeconomical analysis are presented. The second part is devoted to macroeconomical outlooks (France and Europe mainly) on decentralized electricity production (cogeneration, small-scale hydroelectric power plants), to other countries experience (PV systems connected to the grid, cogeneration, etc.) and to price contracts and regulations

  5. Online Simultaneous Hydrogen/Deuterium Exchange of Multitarget Gas-Phase Molecules by Electrospray Ionization Mass Spectrometry Coupled with Gas Chromatography.

    Science.gov (United States)

    Jeong, Eun Sook; Cha, Eunju; Cha, Sangwon; Kim, Sunghwan; Oh, Han Bin; Kwon, Oh-Seung; Lee, Jaeick

    2017-11-21

    In this study, a hydrogen/deuterium (H/D) exchange method using gas chromatography-electrospray ionization/mass spectrometry (GC-ESI/MS) was first investigated as a novel tool for online H/D exchange of multitarget analytes. The GC and ESI source were combined with a homemade heated column transfer line. GC-ESI/MS-based H/D exchange occurs in an atmospheric pressure ion source as a result of reacting the gas-phase analyte eluted from GC with charged droplets of deuterium oxide infused as the ESI spray solvent. The consumption of the deuterated solvent at a flow rate of 2 μL min -1 was more economical than that in online H/D exchange methods reported to date. In-ESI-source H/D exchange by GC-ESI/MS was applied to 11 stimulants with secondary amino or hydroxyl groups. After H/D exchange, the spectra of the stimulants showed unexchanged, partially exchanged, and fully exchanged ions showing various degrees of exchange. The relative abundances corrected for naturally occurring isotopes of the fully exchanged ions of stimulants, except for etamivan, were in the range 24.3-85.5%. Methylephedrine and cyclazodone showed low H/D exchange efficiency under acidic, neutral, and basic spray solvent conditions and nonexchange for etamivan with an acidic phenolic OH group. The in-ESI-source H/D exchange efficiency by GC-ESI/MS was sufficient to determine the number of hydrogen by elucidation of fragmentation from the spectrum. Therefore, this online H/D exchange technique using GC-ESI/MS has potential as an alternative method for simultaneous H/D exchange of multitarget analytes.

  6. Fully decentralized control of a soft-bodied robot inspired by true slime mold.

    Science.gov (United States)

    Umedachi, Takuya; Takeda, Koichi; Nakagaki, Toshiyuki; Kobayashi, Ryo; Ishiguro, Akio

    2010-03-01

    Animals exhibit astoundingly adaptive and supple locomotion under real world constraints. In order to endow robots with similar capabilities, we must implement many degrees of freedom, equivalent to animals, into the robots' bodies. For taming many degrees of freedom, the concept of autonomous decentralized control plays a pivotal role. However a systematic way of designing such autonomous decentralized control system is still missing. Aiming at understanding the principles that underlie animals' locomotion, we have focused on a true slime mold, a primitive living organism, and extracted a design scheme for autonomous decentralized control system. In order to validate this design scheme, this article presents a soft-bodied amoeboid robot inspired by the true slime mold. Significant features of this robot are twofold: (1) the robot has a truly soft and deformable body stemming from real-time tunable springs and protoplasm, the former is used for an outer skin of the body and the latter is to satisfy the law of conservation of mass; and (2) fully decentralized control using coupled oscillators with completely local sensory feedback mechanism is realized by exploiting the long-distance physical interaction between the body parts stemming from the law of conservation of protoplasmic mass. Simulation results show that this robot exhibits highly supple and adaptive locomotion without relying on any hierarchical structure. The results obtained are expected to shed new light on design methodology for autonomous decentralized control system.

  7. Characterization of oligosaccharides with capillary high performance anion exchange chromatography hyphenated to pulsed amperometric detection and ion trap mass spectrometry : application to the analysis of human lysosomal disorders

    NARCIS (Netherlands)

    Bruggink, Cornelis

    The development of a capillary ion chromatograph is described together with a matching desalter. This desalter made it possible to use on-line a mass spectrometer. The mass spectrometer enables partly to characterize carbohydrates eluting from the anion exchange column. This separation technology is

  8. Probing conserved helical modules of portal complexes by mass spectrometry-based hydrogen/deuterium exchange.

    Science.gov (United States)

    Kang, Sebyung; Poliakov, Anton; Sexton, Jennifer; Renfrow, Matthew B; Prevelige, Peter E

    2008-09-05

    The Double-stranded DNA bacteriophage P22 has a ring-shaped dodecameric complex composed of the 84 kDa portal protein subunit that forms the central channel of the phage DNA packaging motor. The overall morphology of the P22 portal complex is similar to that of the portal complexes of Phi29, SPP1, T3, T7 phages and herpes simplex virus. Secondary structure prediction of P22 portal protein and its threading onto the crystal structure of the Phi29 portal complexes suggested that the P22 portal protein complex shares conserved helical modules that were found in the dodecameric interfaces of the Phi29 portal complex. To identify the amino acids involved in intersubunit contacts in the P22 portal ring complexes and validate the threading model, we performed comparative hydrogen/deuterium exchange analysis of monomeric and in vitro assembled portal proteins of P22 and the dodecameric Phi29 portal. Hydrogen/deuterium exchange experiments provided evidence of intersubunit interactions in the P22 portal complex similar to those in the Phi29 portal that map to the regions predicted to be conserved helical modules.

  9. Probing Conserved Helical Modules of Portal Complexes by Mass Spectrometry based Hydrogen/deuterium Exchange

    Science.gov (United States)

    Kang, Sebyung; Poliakov, Anton; Sexton, Jennifer; Renfrow, Matthew B.; Prevelige, Peter E.

    2008-01-01

    The dsDNA bacteriophage P22 has a ring shaped dodecameric complex composed of the 84 kDa portal protein subunit which forms the central channel of the phage’s DNA packaging motor. The overall morphology of the P22 portal complex is similar to that of the portal complexes of Phi29, SPP1, T3, T7 phages and herpes simplex virus. Secondary structure prediction of P22 portal protein and its threading onto the crystal structure of the Phi29 portal complexes suggested that P22 portal protein complex shares conserved helical modules which were found in the dodecameric interfaces of the Phi29 portal complex. To identify the amino acids involved in inter-subunit contacts in the P22 portal ring complexes and validate the threading model, we performed comparative hydrogen/deuterium exchange analysis of monomeric and in vitro assembled portal proteins of P22 and the dodecameric Phi29 portal. Hydrogen/deuterium exchange experiments provided evidence of inter-subunit interactions in the P22 portal complex similar to those in the Phi29 portal which map to the regions predicted to be conserved helical modules. PMID:18621389

  10. Pulsed hydrogen-deuterium exchange mass spectrometry probes conformational changes in amyloid beta (Aβ) peptide aggregation.

    Science.gov (United States)

    Zhang, Ying; Rempel, Don L; Zhang, Jun; Sharma, Anuj K; Mirica, Liviu M; Gross, Michael L

    2013-09-03

    Probing the conformational changes of amyloid beta (Aβ) peptide aggregation is challenging owing to the vast heterogeneity of the resulting soluble aggregates. To investigate the formation of these aggregates in solution, we designed an MS-based biophysical approach and applied it to the formation of soluble aggregates of the Aβ42 peptide, the proposed causative agent in Alzheimer's disease. The approach incorporates pulsed hydrogen-deuterium exchange coupled with MS analysis. The combined approach provides evidence for a self-catalyzed aggregation with a lag phase, as observed previously by fluorescence methods. Unlike those approaches, pulsed hydrogen-deuterium exchange does not require modified Aβ42 (e.g., labeling with a fluorophore). Furthermore, the approach reveals that the center region of Aβ42 is first to aggregate, followed by the C and N termini. We also found that the lag phase in the aggregation of soluble species is affected by temperature and Cu(2+) ions. This MS approach has sufficient structural resolution to allow interrogation of Aβ aggregation in physiologically relevant environments. This platform should be generally useful for investigating the aggregation of other amyloid-forming proteins and neurotoxic soluble peptide aggregates.

  11. Meson exchange second class currents and the neutrino mass in the muon capture by light nuclei

    International Nuclear Information System (INIS)

    Katkhat, Ch.L.

    1988-01-01

    Influence of the Kubodera-Delorme-Rho model parameters (ζ and ξ), the scalar form factor (F s ) and the muonic neutrino rest mass (m νμ ) on the asymmetry coefficient (α μν ) of neutrino emission with respect to the muon spin orientation in the muon capture by light nuclei is analyzed. It is shown, that the mass m νμ , the parameters of ζ and ξ, and the form factor F s may be estimated by studying the coefficient α μν in O -> O, Gamov-Teller, and mixed transitions, respectively

  12. Effective Removal of Nonionic Detergents in Protein Mass Spectrometry, Hydrogen/Deuterium Exchange, and Proteomics

    Czech Academy of Sciences Publication Activity Database

    Rey, M.; Mrázek, H.; Pompach, Petr; Novák, P.; Pelosi, L.; Brandolin, G.; Forest, E.; Havlíček, Vladimír; Man, P.

    2010-01-01

    Roč. 82, č. 12 (2010), s. 5107-5116 ISSN 0003-2700 R&D Projects: GA AV ČR KJB500200612; GA MŠk LC545 Institutional research plan: CEZ:AV0Z50200510 Keywords : mass spectrometry * proteomics * peptides Subject RIV: CE - Biochemistry Impact factor: 5.874, year: 2010

  13. Decentralized Consistent Updates in SDN

    KAUST Repository

    Nguyen, Thanh Dang

    2017-04-10

    We present ez-Segway, a decentralized mechanism to consistently and quickly update the network state while preventing forwarding anomalies (loops and blackholes) and avoiding link congestion. In our design, the centralized SDN controller only pre-computes information needed by the switches during the update execution. This information is distributed to the switches, which use partial knowledge and direct message passing to efficiently realize the update. This separation of concerns has the key benefit of improving update performance as the communication and computation bottlenecks at the controller are removed. Our evaluations via network emulations and large-scale simulations demonstrate the efficiency of ez-Segway, which compared to a centralized approach, improves network update times by up to 45% and 57% at the median and the 99th percentile, respectively. A deployment of a system prototype in a real OpenFlow switch and an implementation in P4 demonstrate the feasibility and low overhead of implementing simple network update functionality within switches.

  14. Integration of decentralized electricity production

    International Nuclear Information System (INIS)

    Tomekova, A.

    2004-01-01

    The project SustelNet also deals with the possibilities of future development of DG sources. Within the project frame a quite general concept of the so-called 'equal field' for centralized and decentralized production was chosen for the better integration of DG. Its aim was to the achieve demanded level on the market in the future term (by 2020). Looking at the problem in the wider context means, that both forms of the production should be admitted on the market on the same conditions. The result of this project is a regulatory map, which actually serves as a definite regulatory strategy for more effective employment of DG sources. On the basis of the national regulatory strategies a proposition of regulatory map for EU will be launched, including some recommendations for European Commission. A few expert papers (scenario of proceeding, benchmarking, economical tools and criteria) are also output of this project. Five member states of the EU and four entering countries have been involved in this project. The final results of this procedure will be presented from April 2004 on international and national conferences, seminaries, or by means of other ways of publicity

  15. Estimation of air-water gas exchange coefficient in a shallow lagoon based on 222Rn mass balance.

    Science.gov (United States)

    Cockenpot, S; Claude, C; Radakovitch, O

    2015-05-01

    The radon-222 mass balance is now commonly used to quantify water fluxes due to Submarine Groundwater Discharge (SGD) in coastal areas. One of the main loss terms of this mass balance, the radon evasion to the atmosphere, is based on empirical equations. This term is generally estimated using one among the many empirical equations describing the gas transfer velocity as a function of wind speed that have been proposed in the literature. These equations were, however, mainly obtained from areas of deep water and may be less appropriate for shallow areas. Here, we calculate the radon mass balance for a windy shallow coastal lagoon (mean depth of 6m and surface area of 1.55*10(8) m(2)) and use these data to estimate the radon loss to the atmosphere and the corresponding gas transfer velocity. We present new equations, adapted to our shallow water body, to express the gas transfer velocity as a function of wind speed at 10 m height (wind range from 2 to 12.5 m/s). When compared with those from the literature, these equations fit particularly well with the one of Kremer et al. (2003). Finally, we emphasize that some gas transfer exchange may always occur, even for conditions without wind. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Hydrothermal alteration and mass exchange in the hornblende latite porphyry, Rico, Colorado

    Science.gov (United States)

    Larson, P.B.; Cunningham, C.G.; Naeser, C.W.

    1994-01-01

    The Rico paleothermal anomaly, southwestern Colorado, records the effects of a large hydrothermal system that was active at 4 Ma. This hydrothermal system produced the deep Silver Creek stockwork Mo deposit, which formed above the anomaly's heat source, and shallower base and precious-metal vein and replacement deposits. A 65 Ma hornblende latite porphyry is present as widespread sills throughout the area and provided a homogenous material that recorded the effects of the hydrothermal system up to 8 km from the center. Hydrothermal alteration in the latite can be divided into a proximal facies which consists of two assemblages, quartz-illite-calcite and chlorite-epidote, and a distal facies which consists of a distinct propylitic assemblage. Temperatures were gradational vertically and laterally in the anomaly, and decreased away from the centra heat source. A convective hydrothermal plume, 3 km wide and at least 2 km high, was present above the stock-work molybdenum deposit and consisted of upwelling, high-temperature fluids that produced the proximal alteration facies. Distal facies alteration was produced by shallower cooler fluids. The most important shallow base and precious-metal vein deposits in the Rico district are at or close to the boundary of the thermal plume. Latite within the plume had a large loss of Na2O, large addition of CaO, and variable SiO2 exchante. Distal propylitized latite samples lost small amounts of Na2O and CaO and exchanged minor variable amounts of SiO2. The edge of the plume is marked by steep Na2O exchange gradients. Na2O exchange throughout the paleothermal anomaly was controlled by the reaction of the albite components in primary plagioclase and alkali feldspars. Initial feldspar alteration in the distal facies was dominated by reaction of the plagioclase, and the initial molar ratio of reactants (alkali feldspar albite component to plagioclase albite component) was 0.35. This ratio of the moles of plagioclase to alkali feldspar

  17. Structural dynamics of soluble chloride intracellular channel protein CLIC1 examined by amide hydrogen-deuterium exchange mass spectrometry.

    Science.gov (United States)

    Stoychev, Stoyan H; Nathaniel, Christos; Fanucchi, Sylvia; Brock, Melissa; Li, Sheng; Asmus, Kyle; Woods, Virgil L; Dirr, Heini W

    2009-09-08

    Chloride intracellular channel protein 1 (CLIC1) functions as an anion channel in plasma and nuclear membranes when its soluble monomeric form converts to an integral-membrane form. The transmembrane region of CLIC1 is located in its thioredoxin-like domain 1, but the mechanism whereby the protein converts to its membrane conformation has yet to be determined. Since channel formation in membranes is enhanced at low pH (5 to 5.5), a condition that is found at the surface of membranes, the structural dynamics of soluble CLIC1 was studied at pH 7 and at pH 5.5 in the absence of membranes by amide hydrogen-deuterium exchange mass spectrometry (DXMS). Rapid hydrogen exchange data indicate that CLIC1 displays a similar core structure at these pH values. Domain 1 is less stable than the all-helical domain 2, and, while the structure of domain 1 remains intact, its conformational flexibility is further increased in an acidic environment (pH 5.5). In the absence of membrane, an acidic environment appears to prime the solution structure of CLIC1 by destabilizing domain 1 in order to lower the activation energy barrier for its conversion to the membrane-insertion conformation. The significantly enhanced H/D-exchange rates at pH 5.5 displayed by two segments (peptides 11-31 and 68-82) could be due to the protonation of acidic residues in salt bridges. One of these segments (peptide 11-31) includes part of the transmembrane region which, in the solution structure, consists of helix alpha1. This helix is intrinsically stable and is most likely retained in the membrane conformation. Strand beta2, another element of the transmembrane region, displays a propensity to form a helical structure and has putative N- and C-capping motifs, suggesting that it too most likely forms a helix in a lipid bilayer.

  18. Supercritical fluid chromatography coupled with in-source atmospheric pressure ionization hydrogen/deuterium exchange mass spectrometry for compound speciation.

    Science.gov (United States)

    Cho, Yunju; Choi, Man-Ho; Kim, Byungjoo; Kim, Sunghwan

    2016-04-29

    An experimental setup for the speciation of compounds by hydrogen/deuterium exchange (HDX) with atmospheric pressure ionization while performing chromatographic separation is presented. The proposed experimental setup combines the high performance supercritical fluid chromatography (SFC) system that can be readily used as an inlet for mass spectrometry (MS) and atmospheric pressure photo ionization (APPI) or atmospheric pressure chemical ionization (APCI) HDX. This combination overcomes the limitation of an approach using conventional liquid chromatography (LC) by minimizing the amount of deuterium solvents used for separation. In the SFC separation, supercritical CO2 was used as a major component of the mobile phase, and methanol was used as a minor co-solvent. By using deuterated methanol (CH3OD), AP HDX was achieved during SFC separation. To prove the concept, thirty one nitrogen- and/or oxygen-containing standard compounds were analyzed by SFC-AP HDX MS. The compounds were successfully speciated from the obtained SFC-MS spectra. The exchange ions were observed with as low as 1% of CH3OD in the mobile phase, and separation could be performed within approximately 20min using approximately 0.24 mL of CH3OD. The results showed that SFC separation and APPI/APCI HDX could be successfully performed using the suggested method. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Effects of sucrose and benzyl alcohol on GCSF conformational dynamics revealed by hydrogen deuterium exchange mass spectrometry.

    Science.gov (United States)

    Zhang, Jun; Banks, Douglas D; He, Feng; Treuheit, Michael J; Becker, Gerald W

    2015-05-01

    Protein stability, one of the major concerns for therapeutic protein development, can be optimized during process development by evaluating multiple formulation conditions. This can be a costly and lengthy procedure where different excipients and storage conditions are tested for their impact on protein stability. A better understanding of the effects of different formulation conditions at the molecular level will provide information on the local interactions within the protein leading to a more rational design of stable and efficacious formulations. In this study, we examined the roles of the excipients, sucrose and benzyl alcohol, on the conformational dynamics of recombinant human granulocyte colony stimulating factor using hydrogen/deuterium exchange coupled with mass spectrometry (HDX-MS). Under physiological pH and temperature, sucrose globally protects the whole molecule from deuterium uptake, whereas benzyl alcohol induces increased deuterium uptake of the regions within the α-helical bundle, with even larger extent. The HDX experiments described were incorporated a set of internal peptides (Zhang et al., 2012. Anal Chem 84:4942-4949) to monitor the differences in intrinsic exchange rates in different formulations. In addition, we discussed the feasibility of implementing HDX-MS with these peptide probes in protein formulation development. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.

  20. Functional unfolding of alpha1-antitrypsin probed by hydrogen-deuterium exchange coupled with mass spectrometry.

    Science.gov (United States)

    Baek, Je-Hyun; Yang, Won Suk; Lee, Cheolju; Yu, Myeong-Hee

    2009-05-01

    The native state of alpha(1)-antitrypsin (alpha(1)AT), a member of the serine protease inhibitor (serpin) family, is considered a kinetically trapped folding intermediate that converts to a more stable form upon complex formation with a target protease. Although previous structural and mutational studies of alpha(1)AT revealed the structural basis of the native strain and the kinetic trap, the mechanism of how the native molecule overcomes the kinetic barrier to reach the final stable conformation during complex formation remains unknown. We hypothesized that during complex formation, a substantial portion of the molecule undergoes unfolding, which we dubbed functional unfolding. Hydrogen-deuterium exchange coupled with ESI-MS was used to analyze this serpin in three forms: native, complexing, and complexed with bovine beta-trypsin. Comparing the deuterium content at the corresponding regions of these three samples, we probed the unfolding of alpha(1)AT during complex formation. A substantial portion of the alpha(1)AT molecule unfolded transiently during complex formation, including not only the regions expected from previous structural studies, such as the reactive site loop, helix F, and the following loop, but also regions not predicted previously, such as helix A, strand 6 of beta-sheet B, and the N terminus. Such unfolding of the native interactions may elevate the free energy level of the kinetically trapped native serpin sufficiently to cross the transition state during complex formation. In the current study, we provide evidence that protein unfolding has to accompany functional execution of the protein molecule.

  1. KRAS G12C Drug Development: Discrimination between Switch II Pocket Configurations Using Hydrogen/Deuterium-Exchange Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Lu, Jia; Harrison, Rane A.; Li, Lianbo; Zeng, Mei; Gondi, Sudershan; Scott, David; Gray, Nathanael S.; Engen, John R.; Westover, Kenneth D. (NEU); (DFCI); (UTSMC); (Harvard-Med)

    2017-09-01

    KRAS G12C, the most common RAS mutation found in non-small-cell lung cancer, has been the subject of multiple recent covalent small-molecule inhibitor campaigns including efforts directed at the guanine nucleotide pocket and separate work focused on an inducible pocket adjacent to the switch motifs. Multiple conformations of switch II have been observed, suggesting that switch II pocket (SIIP) binders may be capable of engaging a range of KRAS conformations. Here we report the use of hydrogen/deuterium-exchange mass spectrometry (HDX MS) to discriminate between conformations of switch II induced by two chemical classes of SIIP binders. We investigated the structural basis for differences in HDX MS using X-ray crystallography and discovered a new SIIP configuration in response to binding of a quinazoline chemotype. These results have implications for structure-guided drug design targeting the RAS SIIP.

  2. Characterizing rapid, activity-linked conformational transitions in proteins via sub-second hydrogen deuterium exchange mass spectrometry.

    Science.gov (United States)

    Resetca, Diana; Wilson, Derek J

    2013-11-01

    This review outlines the application of time-resolved electrospray ionization mass spectrometry (TRESI-MS) and hydrogen-deuterium exchange (HDX) to study rapid, activity-linked conformational transitions in proteins. The method is implemented on a microfluidic chip which incorporates all sample-handling steps required for a 'bottom-up' HDX workflow: a capillary mixer for sub-second HDX labeling, a static mixer for HDX quenching, a microreactor for rapid protein digestion, and on-chip electrospray. By combining short HDX labeling pulses with rapid digestion, this approach provides a detailed characterization of the structural transitions that occur during protein folding, ligand binding, post-translational modification and catalytic turnover in enzymes. This broad spectrum of applications in areas largely inaccessible to conventional techniques means that microfluidics-enabled TRESI-MS/HDX is a unique and powerful approach for investigating the dynamic basis of protein function. © 2013 FEBS.

  3. Gas-phase fragmentation of peptides to increase the spatial resolution of the Hydrogen Exchange Mass Spectrometry experiment

    DEFF Research Database (Denmark)

    Jensen, Pernille Foged; Rand, Kasper Dyrberg

    2016-01-01

    are produced after precursor ion selection and thus do not add complexity to the LC-MS analysis. The key to obtaining optimal spatial resolution in a hydrogen exchange mass spectrometry (HX-MS) experiment is the fragmentation efficiency. This chapter discusses common fragmentation techniques like collision......-induced dissociation (CID) occur with complete Hydrogen-deuterium (H/D) scrambling, while other techniques that induce dissociation on a faster timescale through radical-based fragmentation channels, like electron-capture dissociation (ECD) and electron-transfer dissociation (ETD), occur inherently without H......Generation of overlapping peptides in solution via multiple proteases requires a very high peak capacity for the LC-MS analysis to minimize signal overlap. An inherent advantage of a gas-phase fragmentation step is that the additional gas-phase fragment ions used to sublocalize deuterium...

  4. Probing the Binding Interfaces of Protein Complexes Using Gas-Phase H/D Exchange Mass Spectrometry

    DEFF Research Database (Denmark)

    Mistarz, Ulrik H; Brown, Jeffery M; Haselmann, Kim F

    2016-01-01

    Fast gas-phase hydrogen/deuterium exchange mediated by ND3 gas and measured by mass spectrometry (gas-phase HDX-MS) is a largely unharnessed, fast, and sensitive method for probing primary- and higher-order polypeptide structure. Labeling of heteroatom-bound non-amide hydrogens in a sub...... conditions. Lysozyme ions bound by an oligosaccharide incorporated less deuterium than the unbound ion. Similarly, trypsin ions showed reduced deuterium uptake when bound by the peptide ligand vasopressin. Our results are in good agreement with crystal structures of the native protein complexes......, and illustrate that gas-phase HDX-MS can provide a sensitive and simple approach to measure the number of heteroatom-bound non-amide side-chain hydrogens involved in the binding interface of biologically relevant protein complexes....

  5. Mapping the H-NOX/HK Binding Interface in Vibrio cholerae by Hydrogen/Deuterium Exchange Mass Spectrometry.

    Science.gov (United States)

    Guo, Yirui; Iavarone, Anthony T; Cooper, Matthew M; Marletta, Michael A

    2018-03-20

    Heme-nitric oxide/oxygen binding (H-NOX) proteins are a group of hemoproteins that bind diatomic gas ligands such as nitric oxide (NO) and oxygen (O 2 ). H-NOX proteins typically regulate histidine kinases (HK) located within the same operon. It has been reported that NO-bound H-NOXs inhibit cognate histidine kinase autophosphorylation in bacterial H-NOX/HK complexes; however, a detailed mechanism of NO-mediated regulation of the H-NOX/HK activity remains unknown. In this study, the binding interface of Vibrio cholerae ( Vc) H-NOX/HK complex was characterized by hydrogen/deuterium exchange mass spectrometry (HDX-MS) and further validated by mutagenesis, leading to a new model for NO-dependent kinase inhibition. A conformational change in Vc H-NOX introduced by NO generates a new kinase-binding interface, thus locking the kinase in an inhibitory conformation.

  6. Mg2+ dependence of 70 S ribosomal protein flexibility revealed by hydrogen/deuterium exchange and mass spectrometry.

    Science.gov (United States)

    Yamamoto, Tatsuya; Shimizu, Yoshihiro; Ueda, Takuya; Shiro, Yoshitsugu

    2010-02-19

    The ribosome from Escherichia coli requires a specific concentration of Mg(2+) to maintain the 70 S complex formation and allow protein synthesis, and then the structure must be stable and flexible. How does the ribosome acquire these conflicting factors at the same time? Here, we investigated the hydrogen/deuterium exchange of 52 proteins in the 70 S ribosome, which controlled stability and flexibility under various Mg(2+) concentrations, using mass spectrometry. Many proteins exhibited a sigmoidal curve for Mg(2+) concentration dependence, incorporating more deuterium at lower Mg(2+) concentration. By comparing deuterium incorporation with assembly, we have discovered a typical mechanism of complexes for acquiring both stability and flexibility at the same time. In addition, we got information of the localization of flexibility in ribosomal function by the analysis of related proteins with stalk protein, tRNA, mRNA, and nascent peptide, and demonstrate the relationship between structure, assembly, flexibility, and function of the ribosome.

  7. Analysis of two-phase flow inter-subchannel mass and momentum exchanges by the two-fluid model approach

    Energy Technology Data Exchange (ETDEWEB)

    Ninokata, H. [Tokyo Institute of Technology (Japan); Deguchi, A. [ENO Mathematical Analysis, Tokyo (Japan); Kawahara, A. [Kumamoto Univ., Kumamoto (Japan)

    1995-09-01

    A new void drift model for the subchannel analysis method is presented for the thermohydraulics calculation of two-phase flows in rod bundles where the flow model uses a two-fluid formulation for the conservation of mass, momentum and energy. A void drift model is constructed based on the experimental data obtained in a geometrically simple inter-connected two circular channel test sections using air-water as working fluids. The void drift force is assumed to be an origin of void drift velocity components of the two-phase cross-flow in a gap area between two adjacent rods and to overcome the momentum exchanges at the phase interface and wall-fluid interface. This void drift force is implemented in the cross flow momentum equations. Computational results have been successfully compared to experimental data available including 3x3 rod bundle data.

  8. Study of the mass and energy resolution of the E parallel B charge exchange analyzer for TFTR

    International Nuclear Information System (INIS)

    Kaita, R.; Medley, S.S.

    1979-09-01

    The charge exchange diagnostic for TFTR requires simultaneous multispecie (H + , D + , γ + ) analysis of particles in the energy range of 0.5 - 150 keV. The analyzer design chosen to provide this capability employs a wide gap semi-circular region of superimposed parallel electric and magnetic fields to accomplish mass and energy resolution, respectively. Combined with a large area, multi-anode microchannel plate detector, this arrangement will enable the energy distributions of protons, deuterions, and tritons to be measured concurrently as a function of time during each discharge. A computer simulation program for calculating ion trajectories through the analyzer was written that includes a realistic model of the magnetic and electrostatic fringe fields. This report presents the results of a study of the proposed E parallel B analyzer, and it reveals that the fringe fields are not detrimental to the performance of the analyzer

  9. Energy performance of an innovative liquid desiccant dehumidification system with a counter-flow heat and mass exchanger using potassium formate

    DEFF Research Database (Denmark)

    Jradi, Muhyiddine; Riffat, Saffa

    2014-01-01

    An innovative micro-scale liquid desiccant dehumidification system is numerically investigated. The liquid desiccant dehumidification unit employs a counter-flow low-cost and efficient heat and mass exchange core, improving the thermal performance and eliminating desiccant carryover...... that the dehumidifier effectiveness is directly proportional to the intake air temperature, intake air relative humidity and liquid desiccant flow rate where the effectiveness is inversely proportional to the intake air velocity and the heat exchanger air channel height....

  10. Sustainability evaluation of decentralized electricity generation

    International Nuclear Information System (INIS)

    Karger, Cornelia R.; Hennings, Wilfried

    2009-01-01

    Decentralized power generation is gaining significance in liberalized electricity markets. An increasing decentralization of power supply is expected to make a particular contribution to climate protection. This article investigates the advantages and disadvantages of decentralized electricity generation according to the overall concept of sustainable development. On the basis of a hierarchically structured set of sustainability criteria, four future scenarios for Germany are assessed, all of which describe different concepts of electricity supply in the context of the corresponding social and economic developments. The scenarios are developed in an explorative way according to the scenario method and the sustainability criteria are established by a discursive method with societal actors. The evaluation is carried out by scientific experts. By applying an expanded analytic hierarchy process (AHP), a multicriteria evaluation is conducted that identifies dissent among the experts. The results demonstrate that decentralized electricity generation can contribute to climate protection. The extent to which it simultaneously guarantees security of supply is still a matter of controversy. However, experts agree that technical and economic boundary conditions are of major importance in this field. In the final section, the article discusses the method employed here as well as implications for future decentralized energy supply. (author)

  11. Sustainability evaluation of decentralized electricity generation

    Energy Technology Data Exchange (ETDEWEB)

    Karger, Cornelia R.; Hennings, Wilfried [Research Centre Juelich, Programme Group Humans, Environment, Technology (MUT), 52425 Juelich (Germany)

    2009-04-15

    Decentralized power generation is gaining significance in liberalized electricity markets. An increasing decentralization of power supply is expected to make a particular contribution to climate protection. This article investigates the advantages and disadvantages of decentralized electricity generation according to the overall concept of sustainable development. On the basis of a hierarchically structured set of sustainability criteria, four future scenarios for Germany are assessed, all of which describe different concepts of electricity supply in the context of the corresponding social and economic developments. The scenarios are developed in an explorative way according to the scenario method and the sustainability criteria are established by a discursive method with societal actors. The evaluation is carried out by scientific experts. By applying an expanded analytic hierarchy process (AHP), a multicriteria evaluation is conducted that identifies dissent among the experts. The results demonstrate that decentralized electricity generation can contribute to climate protection. The extent to which it simultaneously guarantees security of supply is still a matter of controversy. However, experts agree that technical and economic boundary conditions are of major importance in this field. In the final section, the article discusses the method employed here as well as implications for future decentralized energy supply. (author)

  12. Two-step ion-exchange chromatographic purification combined with reversed-phase chromatography to isolate C-peptide for mass spectrometric analysis.

    Science.gov (United States)

    Kabytaev, Kuanysh; Durairaj, Anita; Shin, Dmitriy; Rohlfing, Curt L; Connolly, Shawn; Little, Randie R; Stoyanov, Alexander V

    2016-02-01

    A liquid chromatography with mass spectrometry on-line platform that includes the orthogonal techniques of ion exchange and reversed phase chromatography is applied for C-peptide analysis. Additional improvement is achieved by the subsequent application of cation- and anion-exchange purification steps that allow for isolating components that have their isoelectric points in a narrow pH range before final reversed-phase mass spectrometry analysis. The utility of this approach for isolating fractions in the desired "pI window" for profiling complex mixtures is discussed. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Partially decentralized control for ALSTOM gasifier.

    Science.gov (United States)

    Tan, Wen; Lou, Guannan; Liang, Luping

    2011-07-01

    The gasifier plays a key role in the operation of the whole IGCC power plant. It is a typical multivariable control system with strict constraints on the inputs and outputs which makes it very difficult to control. This paper presents a partially decentralized controller design method based on the stabilizer idea. The method only requires identifying some closed-loop transfer functions and solving an H(∞) optimization problem. The final partially decentralized controller is easy to implement and test in practice. Two partially decentralized controllers are designed for the ALSTOM gasifier benchmark problem, and simulation results show that they both meet the design specifications. Copyright © 2011 ISA. Published by Elsevier Ltd. All rights reserved.

  14. Transition-Independent Decentralized Markov Decision Processes

    Science.gov (United States)

    Becker, Raphen; Silberstein, Shlomo; Lesser, Victor; Goldman, Claudia V.; Morris, Robert (Technical Monitor)

    2003-01-01

    There has been substantial progress with formal models for sequential decision making by individual agents using the Markov decision process (MDP). However, similar treatment of multi-agent systems is lacking. A recent complexity result, showing that solving decentralized MDPs is NEXP-hard, provides a partial explanation. To overcome this complexity barrier, we identify a general class of transition-independent decentralized MDPs that is widely applicable. The class consists of independent collaborating agents that are tied up by a global reward function that depends on both of their histories. We present a novel algorithm for solving this class of problems and examine its properties. The result is the first effective technique to solve optimally a class of decentralized MDPs. This lays the foundation for further work in this area on both exact and approximate solutions.

  15. Time resolved investigations on biogenic trace gases exchanges using proton-transfer-reaction mass spectrometry

    International Nuclear Information System (INIS)

    Karl, T.

    2000-02-01

    Volatile organic compounds (VOCs) released from vegetation, including wound-induced VOCs, can have important effects on atmospheric chemistry. The analytical methods for measuring wound-induced VOCs, especially the hexenal family of VOCs (hexenals, hexenols and hexenyl esters) but also compounds like acetaldehyde, are complicated by their chemical instability and the transient nature of their formation after leaf and stem wounding. The goal of this thesis was to assess, quantify and complement our understanding on the origin of tropospheric VOCs. This thesis demonstrates that formation and emission of hexenal family compounds can be monitored on-line using proton-transfer-reaction mass spectrometry (PTR-MS), avoiding the need for preconcentration or chromatography. These measurements revealed the rapid emission of the parent compound, (Z)-3-hexenal, within 1-2 seconds of wounding of leaves from various woody and nonwoody plants, and its metabolites including (E)-2-hexenal, hexenols and hexenyl acetates. Emission of (Z)-3-hexenal from detached, drying leaves averaged 500 μg (gram dry weight)-1. PTR-MS showed to be a useful tool for the analysis of VOC emissions resulting from grazing, herbivory, harvesting and senescing leaves. The release of reactive VOCs during lawn mowing was observed in on-line ambient air measurements in July and August 1998 in the outskirts of Innsbruck. Also obtained were data on emission rates of reactive aldehydes (hexenyl compounds) and other abundant VOCs such as methanol, acetaldehyde and acetone from drying grass in various chamber experiments. Fluxes were measured after cutting of grass using eddy covariance measurements and the micrometeorological gradient method (Obhukov-Similarity-Method). Comparison of data obtained by these different methods showed satisfactory agreement. The highest fluxes for methanol during drying were 5 mg/m2h, for (Z)-3-hexenal 1.5 mg/m2h. Experiments conducted on the Sonnblick Observatory in Fall and Winter

  16. Automated Hydrogen/Deuterium Exchange Electron Transfer Dissociation High Resolution Mass Spectrometry Measured at Single-Amide Resolution

    Science.gov (United States)

    Landgraf, Rachelle R.; Chalmers, Michael J.; Griffin, Patrick R.

    2012-02-01

    Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a well established method for the measurement of solution-phase deuterium incorporation into proteins, which can provide insight into protein conformational mobility. However, most HDX measurements are constrained to regions of the protein where pepsin proteolysis allows detection at peptide resolution. Recently, single-amide resolution deuterium incorporation has been achieved by limiting gas-phase scrambling in the mass spectrometer. This was accomplished by employing a combination of soft ionization and desolvation conditions coupled with the radical-driven fragmentation technique electron transfer dissociation (ETD). Here, a hybrid LTQ-Orbitrap XL is systematically evaluated for its utility in providing single-amide deuterium incorporation for differential HDX analysis of a nuclear receptor upon binding small molecule ligands. We are able to show that instrumental parameters can be optimized to minimize scrambling and can be incorporated into an established and fully automated HDX platform making differential single-amide HDX possible for bottom-up analysis of complex systems. We have applied this system to determine differential single amide resolution HDX data for the peroxizome proliferator activated receptor bound with two ligands of interest.

  17. Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics.

    Science.gov (United States)

    Lento, Cristina; Zhu, Shaolong; Brown, Kerene A; Knox, Ruth; Liuni, Peter; Wilson, Derek J

    2017-04-17

    Intrinsically disordered proteins (IDPs) have long been a challenge to structural biologists due to their lack of stable secondary structure elements. Hydrogen-Deuterium Exchange (HDX) measured at rapid time scales is uniquely suited to detect structures and hydrogen bonding networks that are briefly populated, allowing for the characterization of transient conformers in native ensembles. Coupling of HDX to mass spectrometry offers several key advantages, including high sensitivity, low sample consumption and no restriction on protein size. This technique has advanced greatly in the last several decades, including the ability to monitor HDX labeling times on the millisecond time scale. In addition, by incorporating the HDX workflow onto a microfluidic platform housing an acidic protease microreactor, we are able to localize dynamic properties at the peptide level. In this study, Time-Resolved ElectroSpray Ionization Mass Spectrometry (TRESI-MS) coupled to HDX was used to provide a detailed picture of residual structure in the tau protein, as well as the conformational shifts induced upon hyperphosphorylation.

  18. [Sulfonation modification-assisted enrichment and identification of histidine-containing peptides by strong cation exchange chromatography and mass spectrometry].

    Science.gov (United States)

    Cao, Dong; Zhou, Chunxi; Zhang, Yangjun; Han, Chunguang; Deng, Yulin; Qian, Xiaohong

    2009-03-01

    By the sulfonation at the N-terminal of peptides, the charge state of histidine-containing peptides is different from that of other peptides in pH sulfonated histidine-containing peptides from tryptic digest of proteins by strong cation exchange (SCX) chromatography and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF MS/MS). Using the standard proteins containing histidines as the model, the methodology was evaluated. The results show that sulfonated histidine-containing peptides were efficiently enriched by SCX, and the N-terminal sulfonation of the peptides simplifies the interpretation of the acquired mass spectra and facilitates the sequencing of histidine-containing peptides by producing consecutive and predominant ions in positive mode MS2 spectra, which is thought to be the result of the charge neutralization of b ions by the N-terminal sulfonic acid group. The discrimination of b ions and y ions can greatly enhance the confidence in peptide and subsequent protein identification. It is feasible to isolate and enrich the histidine-containing peptides by using this method which has the potential applications in proteomics.

  19. Decentralization, Local Rights and the Construction of Women's ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Decentralization, Local Rights and the Construction of Women's Citizenship : a Comparative Study in Kenya, Tanzania and Uganda - Phase II. Kenya, Tanzania and Uganda have adopted new land laws, policies and institutional arrangements to accommodate decentralization of land administration and management.

  20. Analysis of transient and hysteresis behavior of cross-flow heat exchangers under variable fluid mass flow rate for data center cooling applications

    International Nuclear Information System (INIS)

    Gao, Tianyi; Murray, Bruce; Sammakia, Bahgat

    2015-01-01

    Effective thermal management of data centers is an important aspect of reducing the energy required for the reliable operation of data processing and communications equipment. Liquid and hybrid (air/liquid) cooling approaches are becoming more widely used in today's large and complex data center facilities. Examples of these approaches include rear door heat exchangers, in-row and overhead coolers and direct liquid cooled servers. Heat exchangers are primary components of liquid and hybrid cooling systems, and the effectiveness of a heat exchanger strongly influences the thermal performance of a cooling system. Characterizing and modeling the dynamic behavior of heat exchangers is important for the design of cooling systems, especially for control strategies to improve energy efficiency. In this study, a dynamic thermal model is solved numerically in order to predict the transient response of an unmixed–unmixed crossflow heat exchanger, of the type that is widely used in data center cooling equipment. The transient response to step and ramp changes in the mass flow rate of both the hot and cold fluid is investigated. Five model parameters are varied over specific ranges to characterize the transient performance. The parameter range investigated is based on available heat exchanger data. The thermal response to the magnitude, time period and initial and final conditions of the transient input functions is studied in detail. Also, the hysteresis associated with the fluid mass flow rate variation is investigated. The modeling results and performance data are used to analyze specific dynamic performance of heat exchangers used in practical data center cooling applications. - Highlights: • The transient performance of a crossflow heat exchanger was modeled and studied. • This study provides design information for data center thermal management. • The time constant metric was used to study the impacts of many variable inputs. • The hysteresis behavior

  1. Decentralization and Economic Growth per capita in Europe

    NARCIS (Netherlands)

    Crucq, Pieter; Hemminga, Hendrik-Jan

    2007-01-01

    In this paper the relationship between decentralization and economic growth is investigated. The focus is on decentralization from the national government to the highest substate level in a country, which we define as regional decentralization. Section 2 discusses the different dimensions of

  2. Time varying controllers in discrete-time decentralized control

    NARCIS (Netherlands)

    Deliu, C.; Deliu, C.; Stoorvogel, Antonie Arij; Saberi, Ali; Roy, Sandip; Malek, Babak

    2009-01-01

    In this paper, we consider the problem of finding a time-varying controller which can stabilize a decentralized discrete-time system. In continuous-time, it was already known that time-varying decentralized controllers can achieve stabilization in cases where time-invariant decentralized controllers

  3. Decentralization of school management to boards of governors

    African Journals Online (AJOL)

    ICL

    structures of decentralization gleaned from the literature, and then discuss some of the elements of decentralization. Theoretical model. The decentralization of ... leadership”. Leadership can only be effective in the Kenyan context if principals work seamlessly with BOGs in school management. Pushpanadham (2006) ...

  4. Online Hydrogen-Deuterium Exchange Traveling Wave Ion Mobility Mass Spectrometry (HDX-IM-MS): a Systematic Evaluation

    Science.gov (United States)

    Cryar, Adam; Groves, Kate; Quaglia, Milena

    2017-06-01

    Hydrogen-deuterium exchange mass spectrometry (HDX-MS) is an important tool for measuring and monitoring protein structure. A bottom-up approach to HDX-MS provides peptide level deuterium uptake values and a more refined localization of deuterium incorporation compared with global HDX-MS measurements. The degree of localization provided by HDX-MS is proportional to the number of peptides that can be identified and monitored across an exchange experiment. Ion mobility spectrometry (IMS) has been shown to improve MS-based peptide analysis of biological samples through increased separation capacity. The integration of IMS within HDX-MS workflows has been commercialized but presently its adoption has not been widespread. The potential benefits of IMS, therefore, have not yet been fully explored. We herein describe a comprehensive evaluation of traveling wave ion mobility integrated within an online-HDX-MS system and present the first reported example of UDMSE acquisition for HDX analysis. Instrument settings required for optimal peptide identifications are described and the effects of detector saturation due to peak compression are discussed. A model system is utilized to confirm the comparability of HDX-IM-MS and HDX-MS uptake values prior to an evaluation of the benefits of IMS at increasing sample complexity. Interestingly, MS and IM-MS acquisitions were found to identify distinct populations of peptides that were unique to the respective methods, a property that can be utilized to increase the spatial resolution of HDX-MS experiments by >60%. [Figure not available: see fulltext.

  5. Conformational dynamics of free and catalytically active thermolysin are indistinguishable by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Liu, Yu-Hong; Konermann, Lars

    2008-06-17

    Conformational dynamics are thought to be a prerequisite for the catalytic activity of enzymes. However, the exact relationship between structural fluctuations and function is not well understood. In this work hydrogen/deuterium exchange (HDX) and electrospray ionization mass spectrometry (ESI-MS) are used for exploring the conformational dynamics of thermolysin. Amide HDX reflects the internal mobility of proteins; regions that undergo frequent unfolding-refolding show faster exchange than segments that are highly stable. Thermolysin is a zinc protease with an active site that is located between two lobes. Substrate turnover is associated with hinge bending that leads to a closed conformation. Product release regenerates the open form, such that steady-state catalysis involves a continuous closing/opening cycle. HDX/ESI-MS with proteolytic peptide mapping in the absence of substrate shows that elements in the periphery of the two lobes are most mobile. A comparison with previous X-ray data suggests that these peripheral regions undergo quite pronounced structural changes during the catalytic cycle. In contrast, active site residues exhibit only a moderate degree of backbone flexibility, and the central zinc appears to be in a fairly rigid environment. The presence of both rigid and moderately flexible elements in the active site may reflect a carefully tuned balance that is required for function. Interestingly, the HDX behavior of catalytically active thermolysin is indistinguishable from that of the free enzyme. This result is consistent with the view that catalytically relevant motions preexist in the resting state and that enzyme function can only be performed within the limitations given by the intrinsic dynamics of the protein. The data presented in this work indicate the prevalence of stochastic elements in the function of thermolysin, rather than supporting a deterministic mechanism.

  6. Application of isotope exchange based mass spectrometry to understand the mechanism of inhibition of sickle hemoglobin polymerization upon oxygenation.

    Science.gov (United States)

    Das, Rajdeep; Mitra, Amrita; Bhat, Vijay; Mandal, Amit Kumar

    2017-07-01

    Sickle hemoglobin (HbS) polymerization initiates in the deoxy state with the binding of hydrophobic patch formed by the isopropyl group of βVal6 residue of a hemoglobin tetramer with the hydrophobic pocket of another tetramer, whose hydrophobic patch binds to the hydrophobic groove of a third molecule. Subsequent elongation of a single stranded polymer followed by the formation of a double strand and finally combination of seven such pairs of double strands results in a fourteen stranded fibrous polymer. Precipitation of this fiber inside the erythrocytes results in sickling of red blood cells. Surprisingly, the polymerization does not occur in the oxy state of HbS. Due to the unavailability of crystal structure of oxy form of HbS, the molecular basis of inhibition of polymerization in the oxy state is unknown to date. In the present study, we have attempted to understand the molecular mechanism of inhibition of polymerization by exploiting the exchange of backbone amide hydrogens of HbS with deuterated solvent. Hydrogen/deuterium exchange kinetics of peptide amide hydrogens of both oxy and deoxy form of HbS were monitored through ESI mass spectrometry. Upon oxygenation changes in the conformational flexibility across different regions of α and β globin chains in the tetrameric HbS molecule were investigated. It was observed that oxygenation led to perturbation in the conformation of several residues around the hydrophobic patch, groove of a tetramer and axial, lateral contacts across the double strands that are involved in HbS polymerization. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Mechanism of Lipid Binding of Human Apolipoprotein E3 by Hydrogen/Deuterium Exchange/Mass Spectrometry and Fluorescence Polarization.

    Science.gov (United States)

    Fabilane, Charina S; Nguyen, Patricia N; Hernandez, Roy V; Nirudodhi, Sasidhar; Duong, Mai; Maier, Claudia S; Narayanaswami, Vasanthy

    2016-01-01

    Human apolipoprotein E3 (apoE3) is an exchangeable apolipoprotein that plays a critical role in maintaining plasma cholesterol/triglyceride homeostasis. The C-terminal (CT) domain of apoE3 (residues 201-299) is composed of amphipathic α-helices C1: W210-S223, C2: V236-E266, and C3: D271-W276, which play a dominant role in mediating high-affinity lipid binding. The objective is to understand the accessibility of the CT domain at the sub-domain level and the mechanistic details regarding lipid-binding interaction. Hydrogen-deuterium exchange coupled to mass spectrometry (HDX/MS) of recombinant wild type (WT) apoE(201-299), chemical-induced unfolding monitored as changes in fluorescence polarization (FP) of labeled apoE(201-299) bearing a probe at specified sites, and lipid binding studies were carried out. HDX/MS revealed that residues towards the C-terminal end of the domain display significantly lower %D uptake compared to those towards the center, suggesting extensive protein-protein interaction in this segment. Functional assays showed that locking apoE(201-299) in an inter-molecular disulfide-bonded state at position 209, 223, 255, or 277 significantly decreases its ability to interact with lipids, especially when tethered towards the ends; this could be restored by reduction. Unfolding studies indicate that the C-terminal end offers less resistance to unfolding compared to the central portion of the domain. Taken together, our data suggest that two dimers of CT domain are juxtaposed around helix C3 leading to apoE3 tetramerization, and that dissociation to monomeric units is a required step in lipid binding, with helix C3 likely seeking stability via lipid interaction prior to helices C1 or C2.

  8. Mapping the Binding Interface in a Noncovalent Size Variant of a Monoclonal Antibody Using Native Mass Spectrometry, Hydrogen-Deuterium Exchange Mass Spectrometry, and Computational Analysis.

    Science.gov (United States)

    Yan, Yuetian; Wei, Hui; Jusuf, Sutjano; Krystek, Stanley R; Chen, Jie; Chen, Guodong; Ludwig, Richard T; Tao, Li; Das, Tapan K

    2017-11-01

    Variants of monoclonal antibody containing an extra light chain have been reported in protein products. Due to potential impact on potency and immunogenicity, it is important to understand the formation mechanism of such variants so that appropriate control strategies can be implemented to assure product quality. In a model monoclonal antibody, we observed a size variant with an extra light chain noncovalently associated with the monomer (later named as "1.2mer"). The interaction between monomer and the extra light chain was characterized by native spray and hydrogen-deuterium exchange mass spectrometry techniques. The goal is to understand the nature of the noncovalent interaction, to map out the interaction interface and regions of potential conformational distortions. In addition, computational modeling was used to aid in binding site identification. The combined results identify the interaction interface to be located in the heavy chain region 38-57 and in the extra light chain region 30-50. To the best of our knowledge, this study is the first to characterize noncovalent interaction of a size variant comprising an antibody monomer and an extra light chain. Structural knowledge generated in this research work is invaluable for process development and construct design of antibody-based biopharmaceuticals. Copyright © 2017. Published by Elsevier Inc.

  9. Selection of greenery plants' group tolerant to mineralized human wastes for their inclusion into intrasystem mass exchange of BTLSS

    Science.gov (United States)

    Tikhomirova, Natalia; Tikhomirov, Alexander A.; Kalacheva, Galina; Ushakova, Sofya; Trifonov, Sergey V.; Pavlova, Anastasiya

    2016-07-01

    A possible way solving the inclusion problem of the human liquid wastes containing sodium chloride into intrasystem mass exchange of bio-technical life support system (BTLSS) is selection of such species of greenery plants that can utilize sodium chloride, be edible for a human and have rather a high productivity. Our previous works showed that salt-accumulating halophyte Salicornia europaea L. was a promising candidate for sodium chloride inclusion into BTLSS mass exchange. However, with the aim of creation of more various human diet possibilities a set of greenery plants cultivated by the water culture method was estimated. Based on preliminary experiments the plants Brassica juncea L., Nasturtium officinale R. Br., Lepidium sativum angustifolia L. and Salicornia europaea L. were chosen as the investigation objects. The nutrient solution for greenery plant's cultivation was obtained after harvesting the wheat plants grown on the solution with mineralized human wastes' addition. The results of the first stage of the investigations carried out showed that plants of Brassica juncea and Lepidium sativum angustifolia are unpromising for their inclusion into BTLSS because of a set of physiological characteristics. On the next stage of investigations, an experimental model of closed ecosystem was created. For that purpose the plants of Salicornia europaea and Nasturtium officinale were introduced in the phototroph unit structure. It was determined that there was deficiency of main macronutrients for Salicornia europaea and Nasturtium officinale including sodium for Salicornia europaea. The deficiency had an effect on plants productivity and on carry-over of sodium from solution by Salicornia europaea. Thus in the future experiment it is necessary to carry out desalinization of solution by Salicornia europaea plants. Nasturtium officinale, that is rich in essential fatty acids and contributes into variety of human vegetable diet is planned to be cultivated by the

  10. Dissecting the Binding Mode of Low Affinity Phage Display Peptide Ligands to Protein Targets by Hydrogen/Deuterium Exchange Coupled to Mass Spectrometry

    DEFF Research Database (Denmark)

    Leurs, Ulrike; Lohse, Brian; Ming, Shonoi A

    2014-01-01

    of hydrogen/deuterium exchange mass spectrometry (HDX-MS) to characterize interactions of low affinity peptides with their cognate protein targets. The HDX-MS workflow was optimized to accurately detect low-affinity peptide-protein interactions by use of ion mobility, electron transfer dissociation, non...

  11. Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins

    DEFF Research Database (Denmark)

    Jensen, Pernille Foged; Comamala Grau, Gerard; Trelle, Morten Beck

    2016-01-01

    and their glycans challenging for most analytical techniques. Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. N-linked glycoproteins however pose a challenge for HDX-MS. HDX...

  12. Assessment of differences in the conformational flexibility of hepatitis B virus core-antigen and e-antigen by hydrogen deuterium exchange-mass spectrometry

    NARCIS (Netherlands)

    Bereszczak, Jessica Z; Watts, Norman R; Wingfield, Paul T; Steven, Alasdair C; Heck, Albert J R

    Hepatitis B virus core-antigen (capsid protein) and e-antigen (an immune regulator) have almost complete sequence identity, yet the dimeric proteins (termed Cp149d and Cp(-10)149d , respectively) adopt quite distinct quaternary structures. Here we use hydrogen deuterium exchange-mass spectrometry

  13. Gas dispersion and bubble-to-emulsion phase mass exchange in a gas-solid bubbling fluidized bed: a computational and experimental study

    NARCIS (Netherlands)

    Patil, D.J.; van Sint Annaland, M.; Kuipers, J.A.M.

    2003-01-01

    Knowledge of gas dispersion and mass exchange between the bubble and the emulsion phases is essential for a correct prediction of the performance of fluidized beds, particularly when catalytic reactions take place. Test cases of single rising bubble and a bubbling fluidized bed operated with a jet

  14. Decentralized Networked Control of Building Structures

    Czech Academy of Sciences Publication Activity Database

    Bakule, Lubomír; Rehák, Branislav; Papík, Martin

    2016-01-01

    Roč. 31, č. 11 (2016), s. 871-886 ISSN 1093-9687 R&D Projects: GA ČR GA13-02149S Institutional support: RVO:67985556 Keywords : decentralized control * networked control * building structures Subject RIV: BC - Control Systems Theory Impact factor: 5.786, year: 2016

  15. GDCluster: A General Decentralized Clustering Algorithm

    NARCIS (Netherlands)

    Mashayekhi, Hoda; Habibi, Jafar; Khalafbeigi, Tania; Voulgaris, Spyros; van Steen, Martinus Richardus

    In many popular applications like peer-to-peer systems, large amounts of data are distributed among multiple sources. Analysis of this data and identifying clusters is challenging due to processing, storage, and transmission costs. In this paper, we propose GDCluster, a general fully decentralized

  16. Women's Political Representation and Participation in Decentralized ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Huairou Commission User

    facilitate people's participation in national development through ensuring sound local level politics. • RC evolved into local councils which then led to the implementation of decentralization through the local government act (1997). • This policy has provided opportunities for women to participate in local leadership from.

  17. Decentralized Development Planning and Fragmentation of ...

    African Journals Online (AJOL)

    Using the Greater Accra Metropolitan Area (GAMA) as a case study, this paper argues that the proliferation of autonomous local government areas within the context of urban sprawl and other challenges have inhibited metropolitan-wide development planning. Keywords: Decentralization; local government; urban growth; ...

  18. Decentralized Sparse Multitask RLS Over Networks

    Science.gov (United States)

    Cao, Xuanyu; Liu, K. J. Ray

    2017-12-01

    Distributed adaptive signal processing has attracted much attention in the recent decade owing to its effectiveness in many decentralized real-time applications in networked systems. Because many natural signals are highly sparse with most entries equal to zero, several decentralized sparse adaptive algorithms have been proposed recently. Most of them is focused on the single task estimation problems, in which all nodes receive data associated with the same unknown vector and collaborate to estimate it. However, many applications are inherently multitask oriented and each node has its own unknown vector different from others'. The related multitask estimation problem benefits from collaborations among the nodes as neighbor nodes usually share analogous properties and thus similar unknown vectors. In this work, we study the distributed sparse multitask recursive least squares (RLS) problem over networks. We first propose a decentralized online alternating direction method of multipliers (ADMM) algorithm for the formulated RLS problem. The algorithm is simplified for easy implementation with closed-form computations in each iteration and low storage requirements. Moreover, to further reduce the complexity, we present a decentralized online subgradient method with low computational overhead. We theoretically analyze the convergence behavior of the proposed subgradient method and derive an error bound related to the network topology and algorithm parameters. The effectiveness of the proposed algorithms is corroborated by numerical simulations and an accuracy-complexity tradeoff between the proposed two algorithms is highlighted.

  19. Group Formation Among Decentralized Autonomous Agents

    NARCIS (Netherlands)

    Ogston, E.F.Y.L.; van Steen, M.R.; Brazier, F.M.

    2004-01-01

    This paper examines a method of clustering within a fully decentralized multi-agent system. Our goal is to group agents with similar objectives or data, as is done in traditional clustering. However, we add the additional constraint that agents must remain in place on a network, instead of first

  20. Metropolitan Schools: Administrative Decentralization vs. Community Control.

    Science.gov (United States)

    Ornstein, Allan C.

    This book is divided into four chapters. The first examines the concepts and issues related to understanding social systems and how the schools can be viewed as a social system. The differences between centralization and decentralization, as well as systems-analysis and management-control approaches are also explored. In the next chapter, we are…

  1. Quotas and Decentralization (Indonesia) | CRDI - Centre de ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Over the past decade, the Government of Indonesia has taken steps to enhance the participation of women in public office. One element of the strategy is decentralization, promoted under the slogan, "local autonomy for people empowerment and welfare." Support for gender mainstreaming was proclaimed and in 2003 an ...

  2. Critical Systems Thinking on Decentralization: the Corporate ...

    African Journals Online (AJOL)

    It proposes more decentralized models of management and outlines a new theory taking a critical systems thinking approach. Corporations are advised to attack and overpower the Corporate Business Virus by re-structuring the dynamics between their headquarters and satellite operations in dealing with the “problem ...

  3. Towards a Decentralized Magnetic Indoor Positioning System

    Directory of Open Access Journals (Sweden)

    Zakaria Kasmi

    2015-12-01

    Full Text Available Decentralized magnetic indoor localization is a sophisticated method for processing sampled magnetic data directly on a mobile station (MS, thereby decreasing or even avoiding the need for communication with the base station. In contrast to central-oriented positioning systems, which transmit raw data to a base station, decentralized indoor localization pushes application-level knowledge into the MS. A decentralized position solution has thus a strong feasibility to increase energy efficiency and to prolong the lifetime of the MS. In this article, we present a complete architecture and an implementation for a decentralized positioning system. Furthermore, we introduce a technique for the synchronization of the observed magnetic field on the MS with the artificially-generated magnetic field from the coils. Based on real-time clocks (RTCs and a preemptive operating system, this method allows a stand-alone control of the coils and a proper assignment of the measured magnetic fields on the MS. A stand-alone control and synchronization of the coils and the MS have an exceptional potential to implement a positioning system without the need for wired or wireless communication and enable a deployment of applications for rescue scenarios, like localization of miners or firefighters.

  4. Decentralized Reinforcement Learning of robot behaviors

    NARCIS (Netherlands)

    Leottau, David L.; Ruiz-del-Solar, Javier; Babuska, R.

    2018-01-01

    A multi-agent methodology is proposed for Decentralized Reinforcement Learning (DRL) of individual behaviors in problems where multi-dimensional action spaces are involved. When using this methodology, sub-tasks are learned in parallel by individual agents working toward a common goal. In

  5. Decentralization Fails Women in Sudan | IDRC - International ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    2010-11-05

    Nov 5, 2010 ... Decentralization – a form of governance required by many donor organizations – is intended to disperse power and resources from the central government to ... Many local institutions have paid little attention to local planning, and those states with fewer resources have a hard time recruiting and retaining ...

  6. Near optimal decentralized H_inf control

    DEFF Research Database (Denmark)

    Stoustrup, J.; Niemann, Hans Henrik

    It is shown that foir a class of decentralized control problems there does not exist a sequence of controllers of bounded order which obtains near optimal control. Neither does there exist an infinity dimentional optimal controller. Using the insight of the line of proof of these results, a heuri...

  7. Quotas and Decentralization (Indonesia) | IDRC - International ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Over the past decade, the Government of Indonesia has taken steps to enhance the participation of women in public office. One element of the strategy is decentralization, promoted under the slogan, "local autonomy for people empowerment and welfare." Support for gender mainstreaming was proclaimed and in 2003 an ...

  8. Decentralized Procurement in Light of Strategic Inventories

    DEFF Research Database (Denmark)

    Arya, Anil; Frimor, Hans; Mittendorf, Brian

    2014-01-01

    and confirmed to be a multifaceted choice. This paper complements existing studies by detailing the trade-offs in the centralization versus decentralization decision in light of firm's decision to cede procurement choices to its individual devisions can help moderate inventory levels and provide a natural salve...

  9. Decentralization, local power and women's rights

    International Development Research Centre (IDRC) Digital Library (Canada)

    Decentralization has changed the political and institutional context for promoting the full and equal rights of citizens in .... to local political, social, economic, and cultural needs and conditions. 13. Research and experience .... Communication, dissemination, and support networks. 52. Disseminate information on quotas, and ...

  10. Decentralization and Diversification in Forest Management Regimes ...

    African Journals Online (AJOL)

    In recent years in many African countries, including Tanzania, there has been a shift of paradigm from centralized and state driven forest management regimes to decentralized and people- centred forest management regime. The inception of a Tanzania forest policy of 1998 resulted in the institutionalization of community ...

  11. Decentralized Decision Making Protocol for Service Composition

    NARCIS (Netherlands)

    Wombacher, Andreas

    Service composition based on state dependent services is a challenge if it done in a decentralized way, that is without a centralized coordinating partner knowing all involved parties. In particular, the challenge is the combination of services to a composite service, such that every party involved

  12. PeerMatcher: Decentralized Partnership Formation

    NARCIS (Netherlands)

    Bozdog, N.V.; Voulgaris, S.; Bal, H.E.; van Halteren, A.

    2015-01-01

    This paper presents PeerMatcher, a fully decentralized algorithm solving the k-clique matching problem. The aim of k-clique matching is to cluster a set of nodes having pairwise weights into k-size groups of maximal total weight. Since solving the problem requires exponential time, PeerMatcher

  13. Quantification of genetically modified soya using strong anion exchange chromatography and time-of-flight mass spectrometry.

    Science.gov (United States)

    Chang, Po-Chih; Reddy, P Muralidhar; Ho, Yen-Peng

    2014-09-01

    Stable-isotope dimethyl labeling was applied to the quantification of genetically modified (GM) soya. The herbicide-resistant gene-related protein 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) was labeled using a dimethyl labeling reagent, formaldehyde-H2 or -D2. The identification and quantification of CP4 EPSPS was performed using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The CP4 EPSPS protein was separated from high abundance proteins using strong anion exchange chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Then, the tryptic peptides from the samples and reference were labeled with formaldehyde-H2 and formaldehyde-D2, respectively. The two labeled pools were mixed and analyzed using MALDI-MS. The data showed a good correlation between the peak ratio of the H- and D-labeled peptides and the GM soya percentages at 0.5, 1, 3, and 5 %, with R (2) of 0.99. The labeling reagents are readily available. The labeling experiments and the detection procedures are simple. The approach is useful for the quantification of GM soya at a level as low as 0.5 %.

  14. Conformational Analysis of Proteins in Highly Concentrated Solutions by Dialysis-Coupled Hydrogen/Deuterium Exchange Mass Spectrometry

    Science.gov (United States)

    Houde, Damian; Nazari, Zeinab E.; Bou-Assaf, George M.; Weiskopf, Andrew S.; Rand, Kasper D.

    2016-04-01

    When highly concentrated, an antibody solution can exhibit unusual behaviors, which can lead to unwanted properties, such as increased levels of protein aggregation and unusually high viscosity. Molecular modeling, along with many indirect biophysical measurements, has suggested that the cause for these phenomena can be due to short range electrostatic and/or hydrophobic protein-protein interactions. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a useful tool for investigating protein conformation, dynamics, and interactions. However, "traditional" continuous dilution labeling HDX-MS experiments have limited utility for the direct analysis of solutions with high concentrations of protein. Here, we present a dialysis-based HDX-MS (di-HDX-MS) method as an alternative HDX-MS labeling format, which takes advantage of passive dialysis rather than the classic dilution workflow. We applied this approach to a highly concentrated antibody solution without dilution or significant sample manipulation, prior to analysis. Such a method could pave the way for a deeper understanding of the unusual behavior of proteins at high concentrations, which is highly relevant for development of biopharmaceuticals in industry.

  15. Ion Mobility Spectrometry-Hydrogen Deuterium Exchange Mass Spectrometry of Anions: Part 2. Assessing Charge Site Location and Isotope Scrambling

    Science.gov (United States)

    Khakinejad, Mahdiar; Ghassabi Kondalaji, Samaneh; Donohoe, Gregory C.; Valentine, Stephen J.

    2016-03-01

    Ion mobility spectrometry (IMS) coupled with gas-phase hydrogen deuterium exchange (HDX)-mass spectrometry (MS) and molecular dynamic simulations (MDS) has been used for structural investigation of anions produced by electrospraying a sample containing a synthetic peptide having the sequence KKDDDDDIIKIIK. In these experiments the potential of the analytical method for locating charge sites on ions as well as for utilizing collision-induced dissociation (CID) to reveal the degree of deuterium uptake within specific amino acid residues has been assessed. For diffuse (i.e., more elongated) [M - 2H]2- ions, decreased deuterium content along with MDS data suggest that the D4 and D6 residues are charge sites, whereas for the more diffuse [M - 3H]3- ions, the data suggest that the D4, D7, and the C-terminus are deprotonated. Fragmentation of mobility-selected, diffuse [M - 2H]2- ions to determine deuterium uptake at individual amino acid residues reveals a degree of deuterium retention at incorporation sites. Although the diffuse [M - 3H]3- ions may show more HD scrambling, it is not possible to clearly distinguish HD scrambling from the expected deuterium uptake based on a hydrogen accessibility model. The capability of the IMS-HDX-MS/MS approach to provide relevant details about ion structure is discussed. Additionally, the ability to extend the approach for locating protonation sites on positively-charged ions is presented.

  16. Ion Mobility Spectrometry-Mass Spectrometry Coupled with Gas-Phase Hydrogen/Deuterium Exchange for Metabolomics Analyses

    Science.gov (United States)

    Maleki, Hossein; Karanji, Ahmad K.; Majuta, Sandra; Maurer, Megan M.; Valentine, Stephen J.

    2018-02-01

    Ion mobility spectrometry-mass spectrometry (IMS-MS) in combination with gas-phase hydrogen/deuterium exchange (HDX) and collision-induced dissociation (CID) is evaluated as an analytical method for small-molecule standard and mixture characterization. Experiments show that compound ions exhibit unique HDX reactivities that can be used to distinguish different species. Additionally, it is shown that gas-phase HDX kinetics can be exploited to provide even further distinguishing capabilities by using different partial pressures of reagent gas. The relative HDX reactivity of a wide variety of molecules is discussed in light of the various molecular structures. Additionally, hydrogen accessibility scoring (HAS) and HDX kinetics modeling of candidate ( in silico) ion structures is utilized to estimate the relative ion conformer populations giving rise to specific HDX behavior. These data interpretation methods are discussed with a focus on developing predictive tools for HDX behavior. Finally, an example is provided in which ion mobility information is supplemented with HDX reactivity data to aid identification efforts of compounds in a metabolite extract.

  17. Dynamics and ligand-induced conformational changes in human prolyl oligopeptidase analyzed by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Tsirigotaki, Alexandra; Elzen, Roos Van; Veken, Pieter Van Der; Lambeir, Anne-Marie; Economou, Anastassios

    2017-05-26

    Prolyl oligopeptidase (PREP) is conserved in many organisms across life. It is involved in numerous processes including brain function and neuropathology, that require more than its strict proteolytic role. It consists of a seven-bladed β-propeller juxtaposed to a catalytic α/β-hydrolase domain. The conformational dynamics of PREP involved in domain motions and the gating mechanism that allows substrate accessibility remain elusive. Here we used Hydrogen Deuterium eXchange Mass Spectrometry (HDX-MS) to derive the first near-residue resolution analysis of global PREP dynamics in the presence or absence of inhibitor bound in the active site. Clear roles are revealed for parts that would be critical for the activation mechanism. In the free state, the inter-domain interface is loose, providing access to the catalytic site. Inhibitor binding "locks" the two domains together exploiting prominent interactions between the loop of the first β-propeller blade and its proximal helix from the α/β-hydrolase domain. Loop A, thought to drive gating, is partially stabilized but remains flexible and dynamic. These findings provide a conformational guide for further dissection of the gating mechanism of PREP, that would impact drug development. Moreover, they offer a structural framework against which to study proteolysis-independent interactions with disordered proteins like α-synuclein involved in neurodegenerative disease.

  18. Hydrogen/Deuterium Exchange Mass Spectrometry Reveals Mechanistic Details of Activation of Nucleoside Diphosphate Kinases by Oligomerization.

    Science.gov (United States)

    Dautant, Alain; Meyer, Philippe; Georgescauld, Florian

    2017-06-13

    Most oligomeric proteins become active only after assembly, but why oligomerization is required to support function is not well understood. Here, we address this question using the wild type (WT) and a destabilized mutant (D93N) of the hexameric nucleoside diphosphate kinase from the pathogen Mycobacterium tuberculosis (Mt-NDPK). The conformational dynamics and oligomeric states of each were analyzed during unfolding and/or folding by hydrogen/deuterium exchange mass spectrometry (HDX-MS) at peptide resolution and by additional biochemical techniques. We found that WT and D93N native hexamers present a stable core and a flexible periphery, the latter being more flexible for the destabilized mutant. Stable but inactive species formed during unfolding of D93N and folding of WT were characterized. For the first time, we show that both of these species are nativelike dimers, each of its monomers having a major subdomain folded, while a minor subdomain (Kpn/α 0 ) remains unfolded. The Kpn/α 0 subdomain, which belongs to the catalytic site, becomes structured only upon hexamerization, explaining why oligomerization is required for NDPK activity. Further HDX-MS studies are necessary to establish the general activation mechanism for other homo-oligomers.

  19. Influence of domain interactions on conformational mobility of the progesterone receptor detected by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Goswami, Devrishi; Callaway, Celetta; Pascal, Bruce D; Kumar, Raj; Edwards, Dean P; Griffin, Patrick R

    2014-07-08

    Structural and functional details of the N-terminal activation function 1 (AF1) of most nuclear receptors are poorly understood due to the highly dynamic intrinsically disordered nature of this domain. A hydrogen/deuterium exchange (HDX) mass-spectrometry-based investigation of TATA box-binding protein (TBP) interaction with various domains of progesterone receptor (PR) demonstrate that agonist-bound PR interaction with TBP via AF1 impacts the mobility of the C-terminal AF2. Results from HDX and other biophysical studies involving agonist- and antagonist-bound full-length PR and isolated PR domains reveal the molecular mechanism underlying synergistic transcriptional activation mediated by AF1 and AF2, dominance of PR-B isoform over PR-A, and the necessity of AF2 for full AF1-mediated transcriptional activity. These results provide a comprehensive picture elaborating the underlying mechanism of PR-TBP interactions as a model for studying nuclear receptor (NR)-transcription factor functional interactions. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Drug binding and resistance mechanism of KIT tyrosine kinase revealed by hydrogen/deuterium exchange FTICR mass spectrometry.

    Science.gov (United States)

    Zhang, Hui-Min; Yu, Xiu; Greig, Michael J; Gajiwala, Ketan S; Wu, Joe C; Diehl, Wade; Lunney, Elizabeth A; Emmett, Mark R; Marshall, Alan G

    2010-04-01

    Mutations of the receptor tyrosine kinase KIT are linked to certain cancers such as gastrointestinal stromal tumors (GISTs). Biophysical, biochemical, and structural studies have provided insight into the molecular basis of resistance to the KIT inhibitors, imatinib and sunitinib. Here, solution-phase hydrogen/deuterium exchange (HDX) and direct binding mass spectrometry experiments provide a link between static structure models and the dynamic equilibrium of the multiple states of KIT, supporting that sunitinib targets the autoinhibited conformation of WT-KIT. The D816H mutation shifts the KIT conformational equilibrium toward the activated state. The V560D mutant exhibits two low energy conformations: one is more flexible and resembles the D816H mutant shifted toward the activated conformation, and the other is less flexible and resembles the wild-type KIT in the autoinhibited conformation. This result correlates with the V560D mutant exhibiting a sensitivity to sunitinib that is less than for WT KIT but greater than for KIT D816H. These findings support the elucidation of the resistance mechanism for the KIT mutants.

  1. High energy efficiency and high power density proton exchange membrane fuel cells: Electrode kinetics and mass transport

    Science.gov (United States)

    Srinivasan, Supramaniam; Velev, Omourtag A.; Parthasathy, Arvind; Manko, David J.; Appleby, A. John

    1991-01-01

    The development of proton exchange membrane (PEM) fuel cell power plants with high energy efficiencies and high power densities is gaining momentum because of the vital need of such high levels of performance for extraterrestrial (space, underwater) and terrestrial (power source for electric vehicles) applications. Since 1987, considerable progress has been made in achieving energy efficiencies of about 60 percent at a current density of 200 mA/sq cm and high power densities (greater than 1 W/sq cm) in PEM fuel cells with high (4 mg/sq cm) or low (0.4 mg/sq cm) platinum loadings in electrodes. The following areas are discussed: (1) methods to obtain these high levels of performance with low Pt loading electrodes - by proton conductor impregnation into electrodes, localization of Pt near front surface; (2) a novel microelectrode technique which yields electrode kinetic parameters for oxygen reduction and mass transport parameters; (3) demonstration of lack of water transport from anode to cathode; (4) modeling analysis of PEM fuel cell for comparison with experimental results and predicting further improvements in performance; and (5) recommendations of needed research and development for achieving the above goals.

  2. Design and experimental analysis of counter-flow heat and mass exchanger incorporating (M-cycle) for evaporative cooling

    Science.gov (United States)

    Khalid, Omar; Butt, Zubair; Tanveer, Waqas; Rao, Hasan Iqbal

    2017-04-01

    In this paper, the functioning of dew-point cooler is improved in terms of its thermal effectiveness. For this reason, a heat and mass exchanger has been designed by using a counter-flow pattern incorporating Maisotsenko cycle (M-cycle) having effective absorbing material called Kraft paper on wet channel side and improved width to height ratio. Experimentation has been performed under various inlet air working parameters such as humidity, velocity and temperature in addition with changing feed water temperature. The results from the experiments specify that the dew-point and the wet-bulb effectiveness is achieved between 67-87 % and 104-120 % respectively. Analysis is performed with temperature variation between 25 and 45 °C at different absolute humidity levels ranging from 14.4 to 18 g/kg, while the inlet air velocity is varied between 0.88 and 1.50 m/s. Thus, the working ability of the improved design has been found 5 % more effective in terms of wet bulb effectiveness as compared to previous counter-flow designs.

  3. Hexicon 2: Automated Processing of Hydrogen-Deuterium Exchange Mass Spectrometry Data with Improved Deuteration Distribution Estimation

    Science.gov (United States)

    Lindner, Robert; Lou, Xinghua; Reinstein, Jochen; Shoeman, Robert L.; Hamprecht, Fred A.; Winkler, Andreas

    2014-06-01

    Hydrogen-deuterium exchange (HDX) experiments analyzed by mass spectrometry (MS) provide information about the dynamics and the solvent accessibility of protein backbone amide hydrogen atoms. Continuous improvement of MS instrumentation has contributed to the increasing popularity of this method; however, comprehensive automated data analysis is only beginning to mature. We present Hexicon 2, an automated pipeline for data analysis and visualization based on the previously published program Hexicon (Lou et al. 2010). Hexicon 2 employs the sensitive NITPICK peak detection algorithm of its predecessor in a divide-and-conquer strategy and adds new features, such as chromatogram alignment and improved peptide sequence assignment. The unique feature of deuteration distribution estimation was retained in Hexicon 2 and improved using an iterative deconvolution algorithm that is robust even to noisy data. In addition, Hexicon 2 provides a data browser that facilitates quality control and provides convenient access to common data visualization tasks. Analysis of a benchmark dataset demonstrates superior performance of Hexicon 2 compared with its predecessor in terms of deuteration centroid recovery and deuteration distribution estimation. Hexicon 2 greatly reduces data analysis time compared with manual analysis, whereas the increased number of peptides provides redundant coverage of the entire protein sequence. Hexicon 2 is a standalone application available free of charge under http://hx2.mpimf-heidelberg.mpg.de.

  4. Hydrogen/Deuterium Exchange Mass Spectrometry Reveals Specific Changes in the Local Flexibility of Plasminogen Activator Inhibitor 1 upon Binding to the Somatomedin B Domain of Vitronectin

    DEFF Research Database (Denmark)

    Trelle, Morten Beck; Hirschberg, Daniel; Jansson, Anna

    2012-01-01

    and increases the thermal stability of the protein dramatically. We have used hydrogen/deuterium exchange mass spectrometry to assess the inherent structural flexibility of PAI-1 and to monitor the changes induced by SMB binding. Our data show that the PAI-1 core consisting of β-sheet B is rather protected...... against exchange with the solvent, while the remainder of the molecule is more dynamic. SMB binding causes a pronounced and widespread stabilization of PAI-1 that is not confined to the binding interface with SMB. We further explored the local structural flexibility in a mutationally stabilized PAI-1...

  5. Assessment of the uncertainties in air mass and pollutants transboundary exchange over the continental part of the EANET region

    Science.gov (United States)

    Gromov, Sergey S.; Trifonova-Yakovleva, Alisa; Gromov, Sergey A.

    2017-04-01

    In this study, we attempt to quantify the uncertainties in air mass exchange in the lower troposphere across two regions of the Russian border in Eastern Siberia and the Russian Far East in 2000-2015. We use meteorological data from long-term air sound data (ASD) on mean layer winds [1] and from the ERA INTERIM re-analysis (EIR) project [2]. Using a transboundary exchange model, we estimate the total and net amounts of air crossing the boundary segments around Irkutsk (IR) and Vladivostok (VL) aerological stations. We compare transport terms derived (i) from the long-term wind statistics based on both ASD and EIR data, and (ii) from integrating 6h meteorological winds from EIR directly over the border segments cells. We find similar wind direction statistics in both meteorological datasets, however EIR favours stronger westerly winds at VL in summer, which results in more often air export from China to Russia in the Far East. There is less agreement on the wind strengths than wind directions between the datasets, with EIR often providing slower wind speeds. The resulting climatic (ASD) and directly (from EIR 6h terms) calculated non-equilibrium (net) transport terms are comparable in orders (tens of million km3/month), however may differ substantially in temporal evolution or/and magnitude. Thus, EIR net transport over the IR segment has similar annual dynamics but is higher by a factor of ˜ 4 (maxima of 3.6 vs. 12 of 106 km3/month in December, respectively). An opposite ratio is derived for the VL segment (average ˜ 6 vs. 13 of 106 km3/month), with a distinct seasonality in the ASD but not in the EIR data. We attribute this discrepancy to the variations in wind direction with altitude, which cannot be resolved in the model fed with the ASD data. Calculated transport in the boundary layer (BL, provided by the EIR) supports this inference. Thus, the BL net transport temporal dynamics differ substantially from that within the 3 km layer, owing to the BL diurnal

  6. (DeCentralization of the Global Informational Ecosystem

    Directory of Open Access Journals (Sweden)

    Johanna Möller

    2017-09-01

    Full Text Available Centralization and decentralization are key concepts in debates that focus on the (antidemocratic character of digital societies. Centralization is understood as the control over communication and data flows, and decentralization as giving it (back to users. Communication and media research focuses on centralization put forward by dominant digital media platforms, such as Facebook and Google, and governments. Decentralization is investigated regarding its potential in civil society, i.e., hacktivism, (encryption technologies, and grass-root technology movements. As content-based media companies increasingly engage with technology, they move into the focus of critical media studies. Moreover, as formerly nationally oriented companies now compete with global media platforms, they share several interests with civil society decentralization agents. Based on 26 qualitative interviews with leading media managers, we investigate (decentralization strategies applied by content-oriented media companies. Theoretically, this perspective on media companies as agents of (decentralization expands (decentralization research beyond traditional democratic stakeholders by considering economic actors within the “global informational ecosystem” (Birkinbine, Gómez, & Wasko, 2017. We provide a three-dimensional framework to empirically investigate (decentralization. From critical media studies, we borrow the (decentralization of data and infrastructures, from media business research, the (decentralization of content distribution.

  7. Histidine hydrogen-deuterium exchange mass spectrometry for probing the microenvironment of histidine residues in dihydrofolate reductase.

    Directory of Open Access Journals (Sweden)

    Masaru Miyagi

    2011-02-01

    Full Text Available Histidine Hydrogen-Deuterium Exchange Mass Spectrometry (His-HDX-MS determines the HDX rates at the imidazole C(2-hydrogen of histidine residues. This method provides not only the HDX rates but also the pK(a values of histidine imidazole rings. His-HDX-MS was used to probe the microenvironment of histidine residues of E. coli dihydrofolate reductase (DHFR, an enzyme proposed to undergo multiple conformational changes during catalysis.Using His-HDX-MS, the pK(a values and the half-lives (t(1/2 of HDX reactions of five histidine residues of apo-DHFR, DHFR in complex with methotrexate (DHFR-MTX, DHFR in complex with MTX and NADPH (DHFR-MTX-NADPH, and DHFR in complex with folate and NADP+ (DHFR-folate-NADP+ were determined. The results showed that the two parameters (pK(a and t(1/2 are sensitive to the changes of the microenvironment around the histidine residues. Although four of the five histidine residues are located far from the active site, ligand binding affected their pK(a, t(1/2 or both. This is consistent with previous observations of ligand binding-induced distal conformational changes on DHFR. Most of the observed pK(a and t(1/2 changes could be rationalized using the X-ray structures of apo-DHFR, DHFR-MTX-NADPH, and DHFR-folate-NADP+. The availability of the neutron diffraction structure of DHFR-MTX enabled us to compare the protonation states of histidine imidazole rings.Our results demonstrate the usefulness of His-HDX-MS in probing the microenvironments of histidine residues within proteins.

  8. Histidine hydrogen-deuterium exchange mass spectrometry for probing the microenvironment of histidine residues in dihydrofolate reductase.

    Science.gov (United States)

    Miyagi, Masaru; Wan, Qun; Ahmad, Md Faiz; Gokulrangan, Giridharan; Tomechko, Sara E; Bennett, Brad; Dealwis, Chris

    2011-02-16

    Histidine Hydrogen-Deuterium Exchange Mass Spectrometry (His-HDX-MS) determines the HDX rates at the imidazole C(2)-hydrogen of histidine residues. This method provides not only the HDX rates but also the pK(a) values of histidine imidazole rings. His-HDX-MS was used to probe the microenvironment of histidine residues of E. coli dihydrofolate reductase (DHFR), an enzyme proposed to undergo multiple conformational changes during catalysis. Using His-HDX-MS, the pK(a) values and the half-lives (t(1/2)) of HDX reactions of five histidine residues of apo-DHFR, DHFR in complex with methotrexate (DHFR-MTX), DHFR in complex with MTX and NADPH (DHFR-MTX-NADPH), and DHFR in complex with folate and NADP+ (DHFR-folate-NADP+) were determined. The results showed that the two parameters (pK(a) and t(1/2)) are sensitive to the changes of the microenvironment around the histidine residues. Although four of the five histidine residues are located far from the active site, ligand binding affected their pK(a), t(1/2) or both. This is consistent with previous observations of ligand binding-induced distal conformational changes on DHFR. Most of the observed pK(a) and t(1/2) changes could be rationalized using the X-ray structures of apo-DHFR, DHFR-MTX-NADPH, and DHFR-folate-NADP+. The availability of the neutron diffraction structure of DHFR-MTX enabled us to compare the protonation states of histidine imidazole rings. Our results demonstrate the usefulness of His-HDX-MS in probing the microenvironments of histidine residues within proteins.

  9. Hydrogen-Deuterium Exchange Mass Spectrometry Reveals Calcium Binding Properties and Allosteric Regulation of Downstream Regulatory Element Antagonist Modulator (DREAM).

    Science.gov (United States)

    Zhang, Jun; Li, Jing; Craig, Theodore A; Kumar, Rajiv; Gross, Michael L

    2017-07-18

    Downstream regulatory element antagonist modulator (DREAM) is an EF-hand Ca 2+ -binding protein that also binds to a specific DNA sequence, downstream regulatory elements (DRE), and thereby regulates transcription in a calcium-dependent fashion. DREAM binds to DRE in the absence of Ca 2+ but detaches from DRE under Ca 2+ stimulation, allowing gene expression. The Ca 2+ binding properties of DREAM and the consequences of the binding on protein structure are key to understanding the function of DREAM. Here we describe the application of hydrogen-deuterium exchange mass spectrometry (HDX-MS) and site-directed mutagenesis to investigate the Ca 2+ binding properties and the subsequent conformational changes of full-length DREAM. We demonstrate that all EF-hands undergo large conformation changes upon calcium binding even though the EF-1 hand is not capable of binding to Ca 2+ . Moreover, EF-2 is a lower-affinity site compared to EF-3 and -4 hands. Comparison of HDX profiles between wild-type DREAM and two EF-1 mutated constructs illustrates that the conformational changes in the EF-1 hand are induced by long-range structural interactions. HDX analyses also reveal a conformational change in an N-terminal leucine-charged residue-rich domain (LCD) remote from Ca 2+ -binding EF-hands. This LCD domain is responsible for the direct interaction between DREAM and cAMP response element-binding protein (CREB) and regulates the recruitment of the co-activator, CREB-binding protein. These long-range interactions strongly suggest how conformational changes transmit the Ca 2+ signal to CREB-mediated gene transcription.

  10. Structural basis of specific interactions of Lp-PLA2 with HDL revealed by hydrogen deuterium exchange mass spectrometry.

    Science.gov (United States)

    Cao, Jian; Hsu, Yuan-Hao; Li, Sheng; Woods, Virgil L; Dennis, Edward A

    2013-01-01

    Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)), specifically Group VIIA PLA(2), is a member of the phospholipase A(2) superfamily and is found mainly associated with LDL and HDL in human plasma. Lp-PLA(2) is considered as a risk factor, a potential biomarker, a target for therapy in the treatment of cardiovascular disease, and evidence suggests that the level of Lp-PLA(2) in plasma is associated with the risk of future cardiovascular and stroke events. The differential location of the enzyme in LDL/HDL lipoproteins has been suggested to affect Lp-PLA(2) function and/or its physiological role and an abnormal distribution of the enzyme may correlate with diseases. Although a mutagenesis study suggested that a surface helix (residues 362-369) mediates the association between Lp-PLA(2) and HDL, the molecular details and mechanism of association has remained unknown. We have now employed hydrogen deuterium exchange mass spectrometry to characterize the interaction between recombinant human Lp-PLA(2) and human HDL. We have found that specific residues 113-120, 192-204, and 360-368 likely mediate HDL binding. In a previous study, we showed that residues 113-120 are important for Lp-PLA(2)-liposome interactions. We now find that residues 192-204 show a decreased deuteration level when Lp-PLA(2) is exposed to apoA-I, but not apoA-II, the most abundant apoproteins in HDL, and additionally, residues 360-368 are only affected by HDL.The results suggest that apoA-I and phospholipid membranes play crucial roles in Lp-PLA(2) localization to HDL.

  11. Effective Application of Bicelles for Conformational Analysis of G Protein-Coupled Receptors by Hydrogen/Deuterium Exchange Mass Spectrometry

    Science.gov (United States)

    Duc, Nguyen Minh; Du, Yang; Thorsen, Thor S.; Lee, Su Youn; Zhang, Cheng; Kato, Hideaki; Kobilka, Brian K.; Chung, Ka Young

    2015-05-01

    G protein-coupled receptors (GPCRs) have important roles in physiology and pathology, and 40% of drugs currently on the market target GPCRs for the treatment of various diseases. Because of their therapeutic importance, the structural mechanism of GPCR signaling is of great interest in the field of drug discovery. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a useful tool for analyzing ligand binding sites, the protein-protein interaction interface, and conformational changes of proteins. However, its application to GPCRs has been limited for various reasons, including the hydrophobic nature of GPCRs and the use of detergents in their preparation. In the present study, we tested the application of bicelles as a means of solubilizing GPCRs for HDX-MS studies. GPCRs (e.g., β2-adrenergic receptor [β2AR], μ-opioid receptor, and protease-activated receptor 1) solubilized in bicelles produced better sequence coverage (greater than 90%) than GPCRs solubilized in n-dodecyl-β-D-maltopyranoside (DDM), suggesting that bicelles are a more effective method of solubilization for HDX-MS studies. The HDX-MS profile of β2AR in bicelles showed that transmembrane domains (TMs) undergo lower deuterium uptake than intracellular or extracellular regions, which is consistent with the fact that the TMs are highly ordered and embedded in bicelles. The overall HDX-MS profiles of β2AR solubilized in bicelles and in DDM were similar except for intracellular loop 3. Interestingly, we detected EX1 kinetics, an important phenomenon in protein dynamics, at the C-terminus of TM6 in β2AR. In conclusion, we suggest the application of bicelles as a useful method for solubilizing GPCRs for conformational analysis by HDX-MS.

  12. Centralization vs. Decentralization in Medical School Libraries

    Science.gov (United States)

    Crawford, Helen

    1966-01-01

    Does the medical school library in the United States operate more commonly under the university library or the medical school administration? University-connected medical school libraries were asked to indicate (a) the source of their budgets, whether from the central library or the medical school, and (b) the responsibility for their acquisitions and cataloging. Returns received from sixtyeight of the seventy eligible institutions showed decentralization to be much the most common: 71 percent of the libraries are funded by their medical schools; 79 percent are responsible for their own acquisitions and processing. The factor most often associated with centralization of both budget and operation is public ownership. Decentralization is associated with service to one or two rather than three or more professional schools. Location of the medical school in a different city from the university is highly favorable to autonomy. Other factors associated with these trends are discussed. PMID:5945568

  13. Centralized, Decentralized, and Hybrid Purchasing Organizations

    DEFF Research Database (Denmark)

    Bals, Lydia; Turkulainen, Virpi

    This paper addresses one of the focal issues in purchasing and supply management – global sourcing – from an organizational design perspective. In particular, we elaborate the traditional classification of global sourcing organization designs into centralized, decentralized, and hybrid models. We...... illustrate with our empirical analysis on global sourcing organization design at Global Chemical Company (GCC, a pseudonym) that revisiting the conventional wisdom about global sourcing organization designs is required; by engaging in a detailed, subfirm level of analysis on the design of the purchasing...... organization we can identify organization designs beyond the classical centralization-decentralization continuum. We also provide explanations for the observed organization design at GCC. The study contributes to research on purchasing and supply management as well as research on organization design....

  14. Mass

    International Nuclear Information System (INIS)

    Quigg, Chris

    2007-01-01

    In the classical physics we inherited from Isaac Newton, mass does not arise, it simply is. The mass of a classical object is the sum of the masses of its parts. Albert Einstein showed that the mass of a body is a measure of its energy content, inviting us to consider the origins of mass. The protons we accelerate at Fermilab are prime examples of Einsteinian matter: nearly all of their mass arises from stored energy. Missing mass led to the discovery of the noble gases, and a new form of missing mass leads us to the notion of dark matter. Starting with a brief guided tour of the meanings of mass, the colloquium will explore the multiple origins of mass. We will see how far we have come toward understanding mass, and survey the issues that guide our research today.

  15. Analyzing Von Neumann machines using decentralized symmetries

    Science.gov (United States)

    Fang, Jie

    2013-10-01

    The artificial intelligence method to e-business is defined not only by the study of fiber-optic cables, but also by the unproven need for vacuum tubes. Given the current status of virtual archetypes, theorists clearly desire the exploration of semaphores, which embodies the compelling principles of cryptoanalysis. We present an algorithm for probabilistic theory (Buck), which we use to disprove that write-back caches can be made decentralized, lossless, and reliable.

  16. WATER MARKETS AND DECENTRALIZED WATER RESOURCES MANAGEMENT

    OpenAIRE

    Easter, K. William; Hearne, Robert R.

    1994-01-01

    Because of its importance and the perceived inability of private sector sources to meet water demands, many countries have depended on the public sector to provide water services for their populations. Yet this has resulted in many inefficient public water projects and in inadequate supplies of good quality and reliable water. Decentralization of water management, including the use of water markets, cannot solve all of the water problems, but it can improve the efficiency of water allocation....

  17. Decentralized estimation and control for power systems

    OpenAIRE

    Singh, Abhinav Kumar

    2014-01-01

    This thesis presents a decentralized alternative to the centralized state-estimation and control technologies used in current power systems. Power systems span over vast geographical areas, and therefore require a robust and reliable communication network for centralized estimation and control. The supervisory control and data acquisition (SCADA) systems provide such a communication architecture and are currently employed for centralized estimation and control of power systems in a static ma...

  18. Dissecting the effect of RNA aptamer binding on the dynamics of plasminogen activator inhibitor 1 using hydrogen/deuterium exchange mass spectrometry

    DEFF Research Database (Denmark)

    Trelle, Morten B; Dupont, Daniel Miotto; Madsen, Jeppe Buur

    2014-01-01

    , about their effects on protein conformation and dynamics. We have employed hydrogen/deuterium exchange (HDX) mass spectrometry to study the effect of RNA aptamers on the structural flexibility of the serpin plasminogen activator inhibitor-1 (PAI-1). The aptamers have characteristic effects...... of the aptamers to PAI-1 is associated with substantial and widespread protection against deuterium uptake in PAI-1. The aptamers induce protection against exchange with the solvent both in the protein-aptamer interface as well as in other specific areas. Interestingly, the aptamers induce substantial protection...... against exchange in α-helices B, C and I. This observation substantiates the relevance of structural instability in this region for transition to the latent state and argues for involvement of flexibility in regions not commonly associated with regulation of latency transition in serpins....

  19. Query Optimizations over Decentralized RDF Graphs

    KAUST Repository

    Abdelaziz, Ibrahim

    2017-05-18

    Applications in life sciences, decentralized social networks, Internet of Things, and statistical linked dataspaces integrate data from multiple decentralized RDF graphs via SPARQL queries. Several approaches have been proposed to optimize query processing over a small number of heterogeneous data sources by utilizing schema information. In the case of schema similarity and interlinks among sources, these approaches cause unnecessary data retrieval and communication, leading to poor scalability and response time. This paper addresses these limitations and presents Lusail, a system for scalable and efficient SPARQL query processing over decentralized graphs. Lusail achieves scalability and low query response time through various optimizations at compile and run times. At compile time, we use a novel locality-aware query decomposition technique that maximizes the number of query triple patterns sent together to a source based on the actual location of the instances satisfying these triple patterns. At run time, we use selectivity-awareness and parallel query execution to reduce network latency and to increase parallelism by delaying the execution of subqueries expected to return large results. We evaluate Lusail using real and synthetic benchmarks, with data sizes up to billions of triples on an in-house cluster and a public cloud. We show that Lusail outperforms state-of-the-art systems by orders of magnitude in terms of scalability and response time.

  20. Towards Automatic Decentralized Control Structure Selection

    DEFF Research Database (Denmark)

    A subtask in integration of design and control of chemical processes is the selection of a control structure. Automating the selection of the control structure enables sequential integration of process and controld esign. As soon as the process is specified or computed, a structure for decentrali......A subtask in integration of design and control of chemical processes is the selection of a control structure. Automating the selection of the control structure enables sequential integration of process and controld esign. As soon as the process is specified or computed, a structure...... for decentralized control is determined automatically, and the resulting decentralized control structure is automatically tuned using standard techniques. Dynamic simulation of the resulting process system gives immediate feedback to the process design engineer regarding practical operability of the process....... The control structure selection problem is formulated as a special MILP employing cost coefficients which are computed using Parseval's theorem combined with RGA and IMC concepts. This approach enables selection and tuning of large-scale plant-wide decentralized controllers through efficient combination...

  1. Towards Automatic Decentralized Control Structure Selection

    DEFF Research Database (Denmark)

    Jørgensen, John Bagterp; Jørgensen, Sten Bay

    2000-01-01

    A subtask in integration of design and control of chemical processes is the selection of a control structure. Automating the selection of the control structure enables sequential integration of process and control design. As soon as the process is specified or computed, a structure for decentrali......A subtask in integration of design and control of chemical processes is the selection of a control structure. Automating the selection of the control structure enables sequential integration of process and control design. As soon as the process is specified or computed, a structure...... for decentralized control is determined automatically, and the resulting decentralized control structure is automatically tuned using standard techniques. Dynamic simulation of the resulting process system gives immediate feedback to the process design engineer regarding practical operability of the process....... The control structure selection problem is formulated as a special MILP employing cost coefficients which are computed using Parseval's theorem combined with RGA and IMC concepts. This approach enables selection and tuning of large-scale plant-wide decentralized controllers through efficient combination...

  2. [Paradoxes of health decentralization policies in Brazil].

    Science.gov (United States)

    Pasche, Dário Frederico; Righi, Liane Beatriz; Thomé, Henrique Inácio; Stolz, Eveline Dischkaln

    2006-12-01

    The constitution of Brazil directs that the country's health system, the Unified Health System (Sistema Unico de Saúde), be politically and administratively decentralized. Nevertheless, handing over competencies, responsibilities, and resources to subnational levels, especially to municipal governments, has been a slow process, lasting almost two decades. Advances have been brought about by the Unified Health System, which, from a analytical perspective, is a public and universal system. Despite that, the decentralization process needs to overcome norms that keep all levels of management dependent on Brazil's federal Government. The subnational levels have consistently faced difficulties in performing their macromanagement functions with autonomy, especially when it comes to financing and to the establishment or organization of health care networks. Boldness and responsibility will be needed to prevent Brazil's health decentralization process from leading to fragmentation. New political agreements between different levels of government, with a reassignment of responsibilities and the enhancement of a culture of technical cooperation, are fundamental requisites to making the Unified Health System have a health policy that is truly public and universal.

  3. Simultaneous determination of 13 carbohydrates using high-performance anion-exchange chromatography coupled with pulsed amperometric detection and mass spectrometry.

    Science.gov (United States)

    Zhao, Dan; Feng, Feng; Yuan, Fei; Su, Jin; Cheng, Yan; Wu, Hanqiu; Song, Kun; Nie, Bo; Yu, Lian; Zhang, Feng

    2017-04-01

    A simple, accurate, and highly sensitive method was developed for the determination of 13 carbohydrates in polysaccharide of Spirulina platensis based on high-performance anion-exchange chromatography coupled with pulsed amperometric detection and mass spectrometry. Samples were extracted with deionized water using ultrasonic-assisted extraction, and the ultrasound-assisted extraction conditions were optimized by Box-Behnken design. Then the extracted polysaccharide was hydrolyzed by adding 1 mol/L trifluoroacetic acid before determination by high-performance anion-exchange chromatography coupled with pulsed amperometric detection and confirmed by high-performance anion-exchange chromatography coupled with mass spectrometry. The high-performance anion-exchange chromatography coupled with pulsed amperometric detection method was performed on a CarboPac PA20 column by gradient elution using deionized water, 0.1 mol/L sodium hydroxide solution, and 0.4 mol/L sodium acetate solution. Excellent linearity was observed in the range of 0.05-10 mg/L. The average recoveries ranged from 80.7 to 121.7%. The limits of detection and limits of quantification for 13 carbohydrates were 0.02-0.10 and 0.2-1.2  μg/kg, respectively. The developed method has been successfully applied to ambient samples, and the results indicated that high-performance anion-exchange chromatography coupled with pulsed amperometric detection and mass spectrometry could provide a rapid and accurate method for the simultaneous determination of carbohydrates. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. A membrane cell for on-line hydrogen/deuterium exchange to study protein folding and protein-protein interactions by mass spectrometry.

    Science.gov (United States)

    Astorga-Wells, Juan; Landreh, Michael; Johansson, Jan; Bergman, Tomas; Jörnvall, Hans

    2011-09-01

    A membrane cell for hydrogen and deuterium exchange on-line with mass spectrometry has been developed to monitor protein-protein interactions and protein conformations. It consists of two channels separated by a semipermeable membrane, where one channel carries the protein sample and the other deuterium oxide. The membrane allows transfer of deuterium oxide into the sample flow. The labeling time is controlled via the flow rate in the sample channel. This cell was validated against three models commonly used in hydrogen-deuterium exchange mass spectrometry: monitoring of folded and unfolded states in a protein, mapping the protein secondary structure at the peptide level, and detection of protein and antibody interactions. The system avoids the conventionally used sample dilution and handling, allowing for potential automation.

  5. A Membrane Cell for On-line Hydrogen/Deuterium Exchange to Study Protein Folding and Protein-Protein Interactions by Mass Spectrometry*

    Science.gov (United States)

    Astorga-Wells, Juan; Landreh, Michael; Johansson, Jan; Bergman, Tomas; Jörnvall, Hans

    2011-01-01

    A membrane cell for hydrogen and deuterium exchange on-line with mass spectrometry has been developed to monitor protein-protein interactions and protein conformations. It consists of two channels separated by a semipermeable membrane, where one channel carries the protein sample and the other deuterium oxide. The membrane allows transfer of deuterium oxide into the sample flow. The labeling time is controlled via the flow rate in the sample channel. This cell was validated against three models commonly used in hydrogen-deuterium exchange mass spectrometry: monitoring of folded and unfolded states in a protein, mapping the protein secondary structure at the peptide level, and detection of protein and antibody interactions. The system avoids the conventionally used sample dilution and handling, allowing for potential automation. PMID:21610101

  6. Approach to Characterization of the Higher Order Structure of Disulfide-Containing Proteins Using Hydrogen/Deuterium Exchange and Top-Down Mass Spectrometry

    OpenAIRE

    Wang, Guanbo; Kaltashov, Igor A.

    2014-01-01

    Top-down hydrogen/deuterium exchange (HDX) with mass spectrometric (MS) detection has recently matured to become a potent biophysical tool capable of providing valuable information on higher order structure and conformational dynamics of proteins at an unprecedented level of structural detail. However, the scope of the proteins amenable to the analysis by top-down HDX MS still remains limited, with the protein size and the presence of disulfide bonds being the two most important limiting fact...

  7. Devolutionary delusions? The effect of decentralization on corruption.

    OpenAIRE

    Ivar Kolstad; Arne Wiig; Vincent Somville

    2014-01-01

    The effect of government decentralization on corruption is theoretically ambiguous. On the one hand, bringing government closer to the people could increase accountability and reduce corruption. On the other hand, decentralization could increase local capture and uncoordinated bribe taking across government levels. This paper estimates the effect of decentralization on experienced corruption, using individual-level bribery data from 36 countries. Crucially, we distinguish between the effect o...

  8. EFFECT OF FISCAL DECENTRALIZATION ON CAPITAL EXPENDITURE, GROWTH, AND WELFARE

    OpenAIRE

    Badrudin, Rudy

    2013-01-01

    This research analyzes the influence of fiscal decentralization on capital expenditure, economic growth, and social welfare of 29 regencies and 6 cities in Central Java Province based on the data of year 2004 to 2008. The method used to analyze the hypotheses is the Partial Least Square. The results showes that fiscal decentralization has no significant effect on capital expenditure; fiscal decentralization has significant effect on economic growth and social welfare; capital expenditure has ...

  9. Peeling the Onion: Why Centralized Control / Decentralized Execution Works

    Science.gov (United States)

    2014-04-01

    March–April 2014 Air & Space Power Journal | 24 Feature Peeling the Onion Why Centralized Control / Decentralized Execution Works Lt Col Alan Docauer...DATES COVERED 00-00-2014 to 00-00-2014 4. TITLE AND SUBTITLE Peeling the Onion : Why Centralized Control / Decentralized Execution Works 5a...Air & Space Power Journal | 25 Docauer Peeling the Onion Feature What Is Centralized Control / Decentralized Execution? Emerging in the aftermath of

  10. Structure and Dynamics of NBD1 from CFTR Characterized Using Crystallography and Hydrogen/Deuterium Exchange Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Lewis, H.A.; Wang, C.; Zhao, X.; Hamuro, Y.; Conners, K.; Kearins, M.C.; Lu, F.; Sauder, J.M.; Molnar, K.S.; Coales, S.J.; Maloney, P.C.; Guggino, W.B.; Wetmore, D.R.; Weber, P.C.; Hunt, J.F. (SGX); (ExSAR); (Cystic); (JHU-MED); (Columbia)

    2012-04-30

    The {Delta}F508 mutation in nucleotide-binding domain 1 (NBD1) of the cystic fibrosis transmembrane conductance regulator (CFTR) is the predominant cause of cystic fibrosis. Previous biophysical studies on human F508 and {Delta}F508 domains showed only local structural changes restricted to residues 509-511 and only minor differences in folding rate and stability. These results were remarkable because {Delta}F508 was widely assumed to perturb domain folding based on the fact that it prevents trafficking of CFTR out of the endoplasmic reticulum. However, the previously reported crystal structures did not come from matched F508 and {Delta}F508 constructs, and the {Delta}F508 structure contained additional mutations that were required to obtain sufficient protein solubility. In this article, we present additional biophysical studies of NBD1 designed to address these ambiguities. Mass spectral measurements of backbone amide {sup 1}H/{sup 2}H exchange rates in matched F508 and {Delta}F508 constructs reveal that {Delta}F508 increases backbone dynamics at residues 509-511 and the adjacent protein segments but not elsewhere in NBD1. These measurements also confirm a high level of flexibility in the protein segments exhibiting variable conformations in the crystal structures. We additionally present crystal structures of a broader set of human NBD1 constructs, including one harboring the native F508 residue and others harboring the {Delta}F508 mutation in the presence of fewer and different solubilizing mutations. The only consistent conformational difference is observed at residues 509-511. The side chain of residue V510 in this loop is mostly buried in all non-{Delta}F508 structures but completely solvent exposed in all {Delta}F508 structures. These results reinforce the importance of the perturbation {Delta}F508 causes in the surface topography of NBD1 in a region likely to mediate contact with the transmembrane domains of CFTR. However, they also suggest that increased

  11. Laser Processed Heat Exchangers

    Data.gov (United States)

    National Aeronautics and Space Administration — The considerable mass of Heat Exchangers (HXs) and coldplates on spacecraft as well as the problematic coatings of the Condensing Heat Exchanger (CHX) are among the...

  12. Closure of mass exchange under use of a vegetable conveyer cultivated on a neutral and soil-like substrates as applied to BLSS

    Science.gov (United States)

    Velitchko, Vladimir; Tikhomirov, Alexander; Ushakova, Sofya

    To increase a closure level of mass exchange processes in bioregenerative life support systems (BLSS) including a human a technology of plants cultivation on a soil-like substrate (SLS) consisting in a gradual decomposition of inedible plants biomass under its addition in the SLS was developed at the Institute of Biophysics SB RAS (Russia). In the given work the effect of periodical introduction of inedible plant biomass in the SLS on plants photosynthetic productivity and on the closure of mass exchange has been analyzed. Thereupon CO2 gas exchange and the certain vegetables' productivity under their cultivation in a conveyor regime on the SLS and on a neutral substrate with reference to the closure of mass exchange processes in BLSS have been studied in this work. The vegetables Raphanus sativus L., Brassica caulorapa L. Daucus carota L. and Beta vulgaris L. being prospective plantsrepresentatives of the BLSS phototrophic unit were taken as the research objects. The SLS was taken as an experiment substrate and an expanded clay aggregate as the control. The changeable Knop solution was used for the control, and an irrigation solution with the SLS extract was used for the experiment. Rapidity dynamics of CO2 consumption showed sharp distinctions of the ‘plants-SLS' system from the ‘plantsexpanded clay aggregate' system connected with the oxidation processes coursing in the SLS. The intensity of CO2 evolution from the SLS on average was 70% of the total plants conveyor's respiration. Thus a balance between the system's respiration and photosynthesis was often determined by the processes coursing in the SLS. Here the sharp CO2 evolution was recorded after introduction of the plants inedible biomass in the SLS. That peak was gradually coming down during 10-14 days after the beginning of every cycle of plants cultivation that was connected with intensification of plants photosynthesis and drop of decomposition intensity of the biomass introduced. Comparative

  13. Carbon source/sink function of a subtropical, eutrophic lake determined from an overall mass balance and a gas exchange and carbon burial balance

    Energy Technology Data Exchange (ETDEWEB)

    Yang Hong [Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China); Environmental Change Research Centre, University College London, London WC1E 6BT (United Kingdom); Xing Yangping [Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China); Graduate School of the Chinese Academy of Sciences, Beijing 100039 (China); Xie Ping [Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China)], E-mail: xieping@ihb.ac.cn; Ni Leyi; Rong Kewen [Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China)

    2008-02-15

    Although studies on carbon burial in lake sediments have shown that lakes are disproportionately important carbon sinks, many studies on gaseous carbon exchange across the water-air interface have demonstrated that lakes are supersaturated with CO{sub 2} and CH{sub 4} causing a net release of CO{sub 2} and CH{sub 4} to the atmosphere. In order to more accurately estimate the net carbon source/sink function of lake ecosystems, a more comprehensive carbon budget is needed, especially for gaseous carbon exchange across the water-air interface. Using two methods, overall mass balance and gas exchange and carbon burial balance, we assessed the carbon source/sink function of Lake Donghu, a subtropical, eutrophic lake, from April 2003 to March 2004. With the overall mass balance calculations, total carbon input was 14 905 t, total carbon output was 4950 t, and net carbon budget was +9955 t, suggesting that Lake Donghu was a great carbon sink. For the gas exchange and carbon burial balance, gaseous carbon (CO{sub 2} and CH{sub 4}) emission across the water-air interface totaled 752 t while carbon burial in the lake sediment was 9477 t. The ratio of carbon emission into the atmosphere to carbon burial into the sediment was only 0.08. This low ratio indicates that Lake Donghu is a great carbon sink. Results showed good agreement between the two methods with both showing Lake Donghu to be a great carbon sink. This results from the high primary production of Lake Donghu, substantive allochthonous carbon inputs and intensive anthropogenic activity. Gaseous carbon emission accounted for about 15% of the total carbon output, indicating that the total output would be underestimated without including gaseous carbon exchange. - Due to high primary production, substantive allochthonous carbon inputs and intensive anthropogenic acitivity, subtropical, eutrophic Lake Donghu is a great carbon sink.

  14. Determination of Equine Cytochrome c Backbone Amide Hydrogen/Deuterium Exchange Rates by Mass Spectrometry Using a Wider Time Window and Isotope Envelope.

    Science.gov (United States)

    Hamuro, Yoshitomo

    2017-03-01

    A new strategy to analyze amide hydrogen/deuterium exchange mass spectrometry (HDX-MS) data is proposed, utilizing a wider time window and isotope envelope analysis of each peptide. While most current scientific reports present HDX-MS data as a set of time-dependent deuteration levels of peptides, the ideal HDX-MS data presentation is a complete set of backbone amide hydrogen exchange rates. The ideal data set can provide single amide resolution, coverage of all exchange events, and the open/close ratio of each amide hydrogen in EX2 mechanism. Toward this goal, a typical HDX-MS protocol was modified in two aspects: measurement of a wider time window in HDX-MS experiments and deconvolution of isotope envelope of each peptide. Measurement of a wider time window enabled the observation of deuterium incorporation of most backbone amide hydrogens. Analysis of the isotope envelope instead of centroid value provides the deuterium distribution instead of the sum of deuteration levels in each peptide. A one-step, global-fitting algorithm optimized exchange rate and deuterium retention during the analysis of each amide hydrogen by fitting the deuterated isotope envelopes at all time points of all peptides in a region. Application of this strategy to cytochrome c yielded 97 out of 100 amide hydrogen exchange rates. A set of exchange rates determined by this approach is more appropriate for a patent or regulatory filing of a biopharmaceutical than a set of peptide deuteration levels obtained by a typical protocol. A wider time window of this method also eliminates false negatives in protein-ligand binding site identification. Graphical Abstract ᅟ.

  15. Determination of Equine Cytochrome c Backbone Amide Hydrogen/Deuterium Exchange Rates by Mass Spectrometry Using a Wider Time Window and Isotope Envelope

    Science.gov (United States)

    Hamuro, Yoshitomo

    2017-03-01

    A new strategy to analyze amide hydrogen/deuterium exchange mass spectrometry (HDX-MS) data is proposed, utilizing a wider time window and isotope envelope analysis of each peptide. While most current scientific reports present HDX-MS data as a set of time-dependent deuteration levels of peptides, the ideal HDX-MS data presentation is a complete set of backbone amide hydrogen exchange rates. The ideal data set can provide single amide resolution, coverage of all exchange events, and the open/close ratio of each amide hydrogen in EX2 mechanism. Toward this goal, a typical HDX-MS protocol was modified in two aspects: measurement of a wider time window in HDX-MS experiments and deconvolution of isotope envelope of each peptide. Measurement of a wider time window enabled the observation of deuterium incorporation of most backbone amide hydrogens. Analysis of the isotope envelope instead of centroid value provides the deuterium distribution instead of the sum of deuteration levels in each peptide. A one-step, global-fitting algorithm optimized exchange rate and deuterium retention during the analysis of each amide hydrogen by fitting the deuterated isotope envelopes at all time points of all peptides in a region. Application of this strategy to cytochrome c yielded 97 out of 100 amide hydrogen exchange rates. A set of exchange rates determined by this approach is more appropriate for a patent or regulatory filing of a biopharmaceutical than a set of peptide deuteration levels obtained by a typical protocol. A wider time window of this method also eliminates false negatives in protein-ligand binding site identification.

  16. Oligonucleotide gas-phase hydrogen/deuterium exchange with D2S in the collision cell of a quadrupole-Fourier transform ion cyclotron resonance mass spectrometer.

    Science.gov (United States)

    Mo, Jingjie; Håkansson, Kristina

    2007-10-15

    We have implemented gas-phase hydrogen/deuterium exchange (HDX) experiments in the external collision cell of a hybrid quadrupole-Fourier transform ion cyclotron resonance mass spectrometer. In this configuration, multiply charged oligonucleotide anions undergo significant exchange with D(2)S at reaction intervals ranging from 0.11 to 60.1 s. For DNA homohexamers, relative exchange rates were dC(6) approximately dA(6) > dG(6) > dT(6), correlating with the gas-phase acidities of nucleobases (C > A > T > G), except for guanine. Our results are consistent with a relay mechanism in which D(2)S interacts with both a backbone phosphate group and a neutral nucleobase through hydrogen bonding. We propose that the faster exchange of polyguanosine compared to polythymidine is due to the larger size of guanine and the orientation of its labile hydrogens, which may result in gas-phase conformations more favorable for forming complexes with D(2)S. Similar trends were observed for RNA homohexamers, although their HDX rates were faster than for DNA, suggesting they can also exchange via another relay process involving the 2'-hydroxyl group. HDX of DNA duplexes further supports the involvement of nucleobase hydrogens because duplexes exchanged slower than their corresponding single strands, presumably due to the intermolecular hydrogen bonds between nucleobases. This work constitutes the first investigation of the mechanisms of oligonucleotide gas-phase HDX. Our results on duplexes show promise for application of this strategy to the characterization of structured nucleic acids.

  17. Decentralized Resource Management in Distributed Computer Systems.

    Science.gov (United States)

    1982-02-01

    Interprocess Communication 14 2.3.2.5 Decentralized Resource Management 15 2.3.3 MicroNet 16 * 2.3.3.1 System Goals and Objectives 16 2.3.3.2 Physical...executive level) is moderately low. 16 Background 2.3.3 MicroNet 2.3.3.1 System Goals and Objectives MicroNet [47] was designed to support multiple...tolerate the loss of nodes, allow for a wide variety of interconnect topologies, and adapt to dynamic variations in loading. The designers of MicroNet

  18. Decentralized Pricing in Minimum Cost Spanning Trees

    DEFF Research Database (Denmark)

    Hougaard, Jens Leth; Moulin, Hervé; Østerdal, Lars Peter

    In the minimum cost spanning tree model we consider decentralized pricing rules, i.e. rules that cover at least the ecient cost while the price charged to each user only depends upon his own connection costs. We de ne a canonical pricing rule and provide two axiomatic characterizations. First......, the canonical pricing rule is the smallest among those that improve upon the Stand Alone bound, and are either superadditive or piece-wise linear in connection costs. Our second, direct characterization relies on two simple properties highlighting the special role of the source cost....

  19. [Secondary Structure of Aβ(1-16) Complexes with Zinc: A Study in the Gas Phase Using Deuterium/Hydrogen Exchange and Ultra-High-Resolution Mass Spectrometry].

    Science.gov (United States)

    Kostyukevich, Yu I; Kononikhin, A S; Indeykina, M I; Popov, I A; Bocharov, K V; Spassky, A I; Kozin, S A; Makarov, A A; Nikolaev, E N

    2017-01-01

    Complexes of peptide fragment 1-16 of beta-amyloid with transition metals play an important role in the development of a broad class of neurodegenerative diseases, which determines the interest in investigating the structures of these complexes. In this work, we have applied the method of the deuterium/hydrogen exchange in combination with ultra-high-resolution mass spectrometry to study conformational changes in (1-16) beta-amyloid peptide induced by binding of zinc(II) atoms. The efficiency of the deuterium/hydrogen exchange depended on the number of zinc atoms bound to the peptide and on the temperature of the ionization source region. Deuterium/hydrogen exchange reactions have been performed directly in the ionization source. The number of exchanges decreased considerably with an increasing numbers of zinc atoms. The relationship has been described with a damped exponential curve, which indicated that the binding of zinc atoms altered the conformation of the peptide ion by making it less open, which limits the access to inner areas of the molecule.

  20. Decentralization, Local Politics and the Construction of Women's ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    This project constitutes the first phase of a comparative study of decentralization, local politics and the construction of women's citizenship in Uganda, Kenya and Tanzania. Researchers will examine the gendered subtexts in the decentralization process with a view to understanding how women's citizenship is constructed ...

  1. Transnational Research on Decentralization in West and Central ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Although decentralization keeps coming up in the political discourse, it is fair to question its viability and effectiveness in the hands of local actors. Following on a pilot project in Ghana and Mali (102772), this research project proposes to analyze, evaluate and compare the process of decentralization underway in six ...

  2. Decentralization : Local Partnerships for Health Services in the ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Cameroon, like most other sub-Saharan African countries, has adopted laws devolving various responsibilities to local administrations. In the local political discourse, decentralization is seen as bringing essential services closer to the users, especially those in greatest need. However, the national decentralization program ...

  3. Decentralization : Local Partnerships for Health Services in the ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    However, the national decentralization program is having a hard time getting on track. In the face of day-to-day difficulties Zenü Network, a nongovernmental organization, would like to make a contribution to this social project. The Network would like to demonstrate that civil society can work with decentralized government ...

  4. Decentralization of health services in India: barriers and facilitating factors.

    Science.gov (United States)

    Kaur, Manmeet; Prinja, Shankar; Singh, Pravin K; Kumar, Rajesh

    2012-01-01

    In India, the process of decentralization of health services started taking shape in the mid-1990s. Systemic reforms envisaged delegation of administrative and financial responsibilities at district level for management of health-care institutions in 23 states of India in 1999. Subsequently, some of these reforms became part of the National Rural Health Mission (NRHM) launched in 2005. This study aims to document the process of decentralization in health services with special reference to the barriers and facilitating factors encountered during formulation and implementation of reform policies. Secondary data were reviewed, health facilities were observed, and semi-structured interviews of the key actors involved in decentralization were carried out in Haryana (India). Political and bureaucratic commitment to reforms was found to be the most important facilitating factor. Orientation training on decentralized administrative structures and performance-based resource distribution were the other important facilitators. Structural changes in administrative procedures led to improvement in the financial management system. Significant improvement in the public health infrastructure was observed. From 2004 to 2008, the state government increased the budget of health sector by nearly 60%. Frequent changes in the top administration at the state level hampered the decentralization process. Districts having a dynamic administrative leadership implemented decentralization more effectively than the rest. Decentralization of financial resources has improved the functioning of health services to some extent. Major policy decisions on decentralization of human resource management, increase in financial allocation, and greater involvement of community in decision-making are required.

  5. The Impact of Fiscal Decentralization on Provision of Quality ...

    African Journals Online (AJOL)

    This article uses panel data from 94 LGAs in Tanzania mainland over the period 2005/06 to 2009/10, to examine the impact of Fiscal Decentralization on education spending and provision of quality education. Using local share of total fiscal expenditure as a proxy for fiscal decentralization, this article concludes that Fiscal ...

  6. Decentralization, local power and women rights: global trends in ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Unicornis

    Women rights and effective decentralization in the MENA: bridging the gaps between local politics, institutional design and gender outcomes. Dr. Mostafa JARI,. Faculty of legal, economic and social sciences. Marrakech Morocco. Decentralization, local power and women rights: global trends in participation, representation ...

  7. Decentralization: A panacea for functional education and national ...

    African Journals Online (AJOL)

    Decentralization of power from the federal government to state and local governments is the way to go, especially in the management of our education system. Education can be best delivered at the state and local government levels. Decentralization of educational management in Nigeria will encourage creativity and ...

  8. Decentralized Consistency Checking in Cross-organizational Workflows

    NARCIS (Netherlands)

    Wombacher, Andreas

    Service Oriented Architectures facilitate loosely coupled composed services, which are established in a decentralized way. One challenge for such composed services is to guarantee consistency, i.e., deadlock-freeness. This paper presents a decentralized approach to consistency checking, which

  9. Application of Transfer Matrix Approach to Modeling and Decentralized Control of Lattice-Based Structures

    Science.gov (United States)

    Cramer, Nick; Swei, Sean Shan-Min; Cheung, Kenny; Teodorescu, Mircea

    2015-01-01

    This paper presents a modeling and control of aerostructure developed by lattice-based cellular materials/components. The proposed aerostructure concept leverages a building block strategy for lattice-based components which provide great adaptability to varying ight scenarios, the needs of which are essential for in- ight wing shaping control. A decentralized structural control design is proposed that utilizes discrete-time lumped mass transfer matrix method (DT-LM-TMM). The objective is to develop an e ective reduced order model through DT-LM-TMM that can be used to design a decentralized controller for the structural control of a wing. The proposed approach developed in this paper shows that, as far as the performance of overall structural system is concerned, the reduced order model can be as e ective as the full order model in designing an optimal stabilizing controller.

  10. Decentralized asset management for collaborative sensing

    Science.gov (United States)

    Malhotra, Raj P.; Pribilski, Michael J.; Toole, Patrick A.; Agate, Craig

    2017-05-01

    There has been increased impetus to leverage Small Unmanned Aerial Systems (SUAS) for collaborative sensing applications in which many platforms work together to provide critical situation awareness in dynamic environments. Such applications require critical sensor observations to be made at the right place and time to facilitate the detection, tracking, and classification of ground-based objects. This further requires rapid response to real-world events and the balancing of multiple, competing mission objectives. In this context, human operators become overwhelmed with management of many platforms. Further, current automated planning paradigms tend to be centralized and don't scale up well to many collaborating platforms. We introduce a decentralized approach based upon information-theory and distributed fusion which enable us to scale up to large numbers of collaborating Small Unmanned Aerial Systems (SUAS) platforms. This is exercised against a military application involving the autonomous detection, tracking, and classification of critical mobile targets. We further show that, based upon monte-carlo simulation results, our decentralized approach out-performs more static management strategies employed by human operators and achieves similar results to a centralized approach while being scalable and robust to degradation of communication. Finally, we describe the limitations of our approach and future directions for our research.

  11. Decentralization and Participatory Rural Development: A Literature Review

    Directory of Open Access Journals (Sweden)

    Muhammad Shakil Ahmad

    2011-12-01

    Full Text Available Most of the developing nations are still struggling for efficient use of their resources. In order to overcome physical and administrative constraints of the development, it is necessary to transfer the power from the central government to local authorities. Distribution of power from improves the management of resources and community participation which is considered key to sustainable development. Advocates of decentralization argue that decentralized government is source to improve community participation in rural development. Decentralized government is considered more responsive towards local needs and development of poor peoples. There are many obstacles to expand the citizen participation in rural areas. There are many approaches for participatory development but all have to face the same challenges. Current paper highlights the literature about Decentralization and participatory rural development. Concept and modalities of Decentralization, dimensions of participation, types of rural participation and obstacles to participation are also the part of this paper.

  12. Pinpointing changes in higher-order protein structure by hydrogen/deuterium exchange coupled to electron transfer dissociation mass spectrometry

    DEFF Research Database (Denmark)

    Rand, Kasper Dyrberg

    2013-01-01

    This Feature describes the use of electron transfer dissociation (ETD) to analyze the hydrogen/deuterium exchange (HDX) of proteins at increased spatial resolution down to the level of individual residues. A practical overview of how to couple ETD to the classical bottom-up HDX-MS workflow is given...

  13. Rapid identification of phase I and II metabolites of artemisinin antimalarials using LTQ-Orbitrap hybrid mass spectrometer in combination with online hydrogen/deuterium exchange technique.

    Science.gov (United States)

    Liu, Tian; Du, Fuying; Wan, Yakun; Zhu, Fanping; Xing, Jie

    2011-08-01

    Artemisinin drugs have become the first-line antimalarials in areas of multi-drug resistance. However, monotherapy with artemisinin drugs results in comparatively high recrudescence rates. Autoinduction of CYP-mediated metabolism, resulting in reduced exposure, has been supposed to be the underlying mechanism. To better understand the autoinduction of artemisinin drugs, we evaluated the biotransformation of artemisinin, also known as Qing-hao-su (QHS), and its active derivative dihydroartemisinin (DHA) in vitro and in vivo, using LTQ-Orbitrap hybrid mass spectrometer in conjunction with online hydrogen (H)/deuterium (D) exchange high-resolution (HR)-LC/MS (mass spectrometry) for rapid structural characterization. The LC separation was improved allowing the separation of QHS parent drugs and their metabolites from their diastereomers. Thirteen phase I metabolites of QHS have been identified in liver microsomal incubates, rat urine, bile and plasma, including six deoxyhydroxylated metabolites, five hydroxylated metabolites, one dihydroxylated metabolite and deoxyartemisinin. Twelve phase II metabolites of QHS were detected in rat bile, urine and plasma. DHA underwent similar metabolic pathways, and 13 phase I metabolites and 3 phase II metabolites were detected. Accurate mass data were obtained in both full-scan and MS/MS mode to support assignments of metabolite structures. Online H/D exchange LC-HR/MS experiments provided additional evidence in differentiating deoxydihydroxylated metabolites from mono-hydroxylated metabolites. The results showed that the main phase I metabolites of artemisinin drugs are hydroxylated and deoxyl products, and they will undergo subsequent phase II glucuronidation processes. This study also demonstrated the effectiveness of online H/D exchange LC-HR/MS(n) technique in rapid identification of drug metabolites. Copyright © 2011 John Wiley & Sons, Ltd.

  14. Using hydrogen/deuterium exchange mass spectrometry to define the specific interactions of the phospholipase A2 superfamily with lipid substrates, inhibitors, and membranes.

    Science.gov (United States)

    Cao, Jian; Burke, John E; Dennis, Edward A

    2013-01-18

    The phospholipase A(2) (PLA(2)) superfamily consists of 16 groups and many subgroups and constitutes a diverse set of enzymes that have a common catalytic activity due to convergent evolution. However, different PLA(2) types have unique three-dimensional structures and catalytic residues as well as specific tissue localization and distinct biological functions. Understanding how the different PLA(2) enzymes associate with phospholipid membranes, specific phospholipid substrate molecules, and inhibitors on a molecular basis has advanced in recent years due to the introduction of hydrogen/deuterium exchange mass spectrometry. Its theory, practical considerations, and application to understanding PLA(2)/membrane interactions are addressed.

  15. Probing the Conformational and Functional Consequences of Disulfide Bond Engineering in Growth Hormone by Hydrogen-Deuterium Exchange Mass Spectrometry Coupled to Electron Transfer Dissociation

    DEFF Research Database (Denmark)

    Seger, Signe T; Breinholt, Jens; Faber, Johan H

    2015-01-01

    Human growth hormone (hGH), and its receptor interaction, is essential for cell growth. To stabilize a flexible loop between helices 3 and 4, while retaining affinity for the hGH receptor, we have engineered a new hGH variant (Q84C/Y143C). Here, we employ hydrogen-deuterium exchange mass...... ranging effects, stabilizing a short α-helix quite distant from the mutation sites, but also rendering a part of the α-helical hGH core slightly more dynamic. In the regions where the hGH variant exhibits a different deuterium uptake than the wild type protein, electron transfer dissociation (ETD...

  16. Using Hydrogen/Deuterium Exchange Mass Spectrometry to Define the Specific Interactions of the Phospholipase A2 Superfamily with Lipid Substrates, Inhibitors, and Membranes*

    Science.gov (United States)

    Cao, Jian; Burke, John E.; Dennis, Edward A.

    2013-01-01

    The phospholipase A2 (PLA2) superfamily consists of 16 groups and many subgroups and constitutes a diverse set of enzymes that have a common catalytic activity due to convergent evolution. However, different PLA2 types have unique three-dimensional structures and catalytic residues as well as specific tissue localization and distinct biological functions. Understanding how the different PLA2 enzymes associate with phospholipid membranes, specific phospholipid substrate molecules, and inhibitors on a molecular basis has advanced in recent years due to the introduction of hydrogen/deuterium exchange mass spectrometry. Its theory, practical considerations, and application to understanding PLA2/membrane interactions are addressed. PMID:23209293

  17. Dynamics and ligand-induced conformational changes in human prolyl oligopeptidase analyzed by hydrogen/deuterium exchange mass spectrometry

    OpenAIRE

    Tsirigotaki, Alexandra; Elzen, van, Roos; Veken, van der, Pieter; Lambeir, Anne-Marie; Economou, Anastassios

    2017-01-01

    Abstract: Prolyl oligopeptidase (PREP) is conserved in many organisms across life. It is involved in numerous processes including brain function and neuropathology, that require more than its strict proteolytic role. It consists of a seven-bladed beta-propeller juxtaposed to a catalytic alpha/beta-hydrolase domain. The conformational dynamics of PREP involved in domain motions and the gating mechanism that allows substrate accessibility remain elusive. Here we used Hydrogen Deuterium eXchange...

  18. Dissecting the effect of RNA aptamer binding on the dynamics of plasminogen activator inhibitor 1 using hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Trelle, Morten B; Dupont, Daniel M; Madsen, Jeppe B; Andreasen, Peter A; Jørgensen, Thomas J D

    2014-01-17

    RNA aptamers, selected from large synthetic libraries, are attracting increasing interest as protein ligands, with potential uses as prototype pharmaceuticals, conformational probes, and reagents for specific quantification of protein levels in biological samples. Very little is known, however, about their effects on protein conformation and dynamics. We have employed hydrogen/deuterium exchange (HDX) mass spectrometry to study the effect of RNA aptamers on the structural flexibility of the serpin plasminogen activator inhibitor-1 (PAI-1). The aptamers have characteristic effects on the biochemical properties of PAI-1. In particular, they are potent inhibitors of the structural transition of PAI-1 from the active state to the inactive, so-called latent state. This transition is one of the largest conformational changes of a folded protein domain without covalent modification. Binding of the aptamers to PAI-1 is associated with substantial and widespread protection against deuterium uptake in PAI-1. The aptamers induce protection against exchange with the solvent both in the protein-aptamer interface as well as in other specific areas. Interestingly, the aptamers induce substantial protection against exchange in α-helices B, C and I. This observation substantiates the relevance of structural instability in this region for transition to the latent state and argues for involvement of flexibility in regions not commonly associated with regulation of latency transition in serpins.

  19. Energy and air emission implications of a decentralized wastewater system

    International Nuclear Information System (INIS)

    Shehabi, Arman; Stokes, Jennifer R; Horvath, Arpad

    2012-01-01

    Both centralized and decentralized wastewater systems have distinct engineering, financial and societal benefits. This paper presents a framework for analyzing the environmental effects of decentralized wastewater systems and an evaluation of the environmental impacts associated with two currently operating systems in California, one centralized and one decentralized. A comparison of energy use, greenhouse gas emissions and criteria air pollutants from the systems shows that the scale economies of the centralized plant help lower the environmental burden to less than a fifth of that of the decentralized utility for the same volume treated. The energy and emission burdens of the decentralized plant are reduced when accounting for high-yield wastewater reuse if it supplants an energy-intensive water supply like a desalination one. The centralized facility also reduces greenhouse gases by flaring methane generated during the treatment process, while methane is directly emitted from the decentralized system. The results are compelling enough to indicate that the life-cycle environmental impacts of decentralized designs should be carefully evaluated as part of the design process. (letter)

  20. Decentralized energy supply on the liberalized market

    International Nuclear Information System (INIS)

    Pauli, H.

    1999-01-01

    Starting in 2001, the electricity market is to be progressively liberalized. The process will be completed by the year 2006. What role will decentralized power generation using combined cycle power plants play on a liberalized market ? The background conditions are essentially favourable: both the new energy act, which has been in force since 1 January 1999, and the planned energy levy suggest that this technology will become increasingly widespread. In addition, the price trend for combined cycle plants components together with low energy costs are having a favourable impact. On the other hand, great uncertainty is being created by the process of liberalization and the current flood of investments in power generation. However, electricity supply is unlikely to be in surplus for long in a context of sustained economic growth. (author)

  1. Load scheduling for decentralized CHP plants

    DEFF Research Database (Denmark)

    Nielsen, Henrik Aalborg, orlov 31.07.2008; Madsen, Henrik; Nielsen, Torben Skov

    be obtained. Furthermore, we believe that all relevant forecasting methods are far too complicated to allow for this integration; both uncertainties originating from the dependence of heat load on climate and from meteorological forecasts need to be taken into account. Instead we suggest that the decision......This report considers load scheduling for decentralized combined heat and power plants where the revenue from selling power to the transmission company and the fuel cost may be time-varying. These plants produce both heat and power with a fixed ratio between these outputs. A heat storage facility...... is used to be able to deviate from this restriction. The load scheduling must be performed with only approximate knowledge about the future. At present in Denmark this uncertainty is only associated with the heat demand, but in the future revenues of produced energy and the fuel costs might also...

  2. Influence of cardiac decentralization on cardioprotection.

    Directory of Open Access Journals (Sweden)

    John G Kingma

    Full Text Available The role of cardiac nerves on development of myocardial tissue injury after acute coronary occlusion remains controversial. We investigated whether acute cardiac decentralization (surgical modulates coronary flow reserve and myocardial protection in preconditioned dogs subject to ischemia-reperfusion. Experiments were conducted on four groups of anesthetised, open-chest dogs (n = 32: 1- controls (CTR, intact cardiac nerves, 2- ischemic preconditioning (PC; 4 cycles of 5-min IR, 3- cardiac decentralization (CD and 4- CD+PC; all dogs underwent 60-min coronary occlusion and 180-min reperfusion. Coronary blood flow and reactive hyperemic responses were assessed using a blood volume flow probe. Infarct size (tetrazolium staining was related to anatomic area at risk and coronary collateral blood flow (microspheres in the anatomic area at risk. Post-ischemic reactive hyperemia and repayment-to-debt ratio responses were significantly reduced for all experimental groups; however, arterial perfusion pressure was not affected. Infarct size was reduced in CD dogs (18.6 ± 4.3; p = 0.001, data are mean ± 1 SD compared to 25.2 ± 5.5% in CTR dogs and was less in PC dogs as expected (13.5 ± 3.2 vs. 25.2 ± 5.5%; p = 0.001; after acute CD, PC protection was conserved (11.6 ± 3.4 vs. 18.6 ± 4.3%; p = 0.02. In conclusion, our findings provide strong evidence that myocardial protection against ischemic injury can be preserved independent of extrinsic cardiac nerve inputs.

  3. Hydrogen exchange

    DEFF Research Database (Denmark)

    Jensen, Pernille Foged; Rand, Kasper Dyrberg

    2016-01-01

    Hydrogen exchange (HX) monitored by mass spectrometry (MS) is a powerful analytical method for investigation of protein conformation and dynamics. HX-MS monitors isotopic exchange of hydrogen in protein backbone amides and thus serves as a sensitive method for probing protein conformation...... and dynamics along the entire protein backbone. This chapter describes the exchange of backbone amide hydrogen which is highly quenchable as it is strongly dependent on the pH and temperature. The HX rates of backbone amide hydrogen are sensitive and very useful probes of protein conformation......, as they are distributed along the polypeptide backbone and form the fundamental hydrogen-bonding networks of basic secondary structure. The effect of pressure on HX in unstructured polypeptides (poly-dl-lysine and oxidatively unfolded ribonuclease A) and native folded proteins (lysozyme and ribonuclease A) was evaluated...

  4. Optimization and application of atmospheric pressure chemical and photoionization hydrogen-deuterium exchange mass spectrometry for speciation of oxygen-containing compounds.

    Science.gov (United States)

    Acter, Thamina; Kim, Donghwi; Ahmed, Arif; Jin, Jang Mi; Yim, Un Hyuk; Shim, Won Joon; Kim, Young Hwan; Kim, Sunghwan

    2016-05-01

    This paper presents a detailed investigation of the feasibility of optimized positive and negative atmospheric pressure chemical ionization (APCI) mass spectrometry (MS) and atmospheric pressure photoionization (APPI) MS coupled to hydrogen-deuterium exchange (HDX) for structural assignment of diverse oxygen-containing compounds. The important parameters for optimization of HDX MS were characterized. The optimized techniques employed in the positive and negative modes showed satisfactory HDX product ions for the model compounds when dichloromethane and toluene were employed as a co-solvent in APCI- and APPI-HDX, respectively. The evaluation of the mass spectra obtained from 38 oxygen-containing compounds demonstrated that the extent of the HDX of the ions was structure-dependent. The combination of information provided by different ionization techniques could be used for better speciation of oxygen-containing compounds. For example, (+) APPI-HDX is sensitive to compounds with alcohol, ketone, or aldehyde substituents, while (-) APPI-HDX is sensitive to compounds with carboxylic functional groups. In addition, the compounds with alcohol can be distinguished from other compounds by the presence of exchanged peaks. The combined information was applied to study chemical compositions of degraded oils. The HDX pattern, double bond equivalent (DBE) distribution, and previously reported oxidation products were combined to predict structures of the compounds produced from oxidation of oil. Overall, this study shows that APCI- and APPI-HDX MS are useful experimental techniques that can be applied for the structural analysis of oxygen-containing compounds.

  5. Solid-State Hydrogen-Deuterium Exchange Mass Spectrometry: Correlation of Deuterium Uptake and Long-Term Stability of Lyophilized Monoclonal Antibody Formulations.

    Science.gov (United States)

    Moorthy, Balakrishnan S; Zarraga, Isidro E; Kumar, Lokesh; Walters, Benjamin T; Goldbach, Pierre; Topp, Elizabeth M; Allmendinger, Andrea

    2018-01-02

    Solid state hydrogen-deuterium exchange with mass spectrometric analysis (ssHDX-MS) has been used to assess protein conformation and matrix interactions in lyophilized solids. ssHDX-MS metrics have been previously correlated to the formation of aggregates of lyophilized myoglobin on storage. Here, ssHDX-MS was applied to lyophilized monoclonal antibody (mAb) formulations and correlated to their long-term stability. After exposing lyophilized samples to D 2 O(g), the amount of deuterium incorporated at various time points was determined by mass spectrometry for four different lyophilized mAb formulations. Hydrogen-deuterium exchange data were then correlated with mAb aggregation and chemical degradation, which was obtained in stability studies of >2.5 years. Deuterium uptake on ssHDX-MS of four lyophilized mAb formulations determined at the initial time point prior to storage in the dry state was directly and strongly correlated with the extent of aggregation and chemical degradation during storage. Other measures of physical and chemical properties of the solids were weakly or poorly correlated with stability. The data demonstrate, for the first time, that ssHDX-MS results are highly correlated with the stability of lyophilized mAb formulations. The findings thus suggest that ssHDX-MS can be used as an early read-out of differences in long-term stability between formulations helping to accelerate formulation screening and selection.

  6. Combination of decentralized waste drying and SSF techniques for household biowaste minimization and ethanol production.

    Science.gov (United States)

    Sotiropoulos, A; Vourka, I; Erotokritou, A; Novakovic, J; Panaretou, V; Vakalis, S; Thanos, T; Moustakas, K; Malamis, D

    2016-06-01

    The results of the demonstration of an innovative household biowaste management and treatment scheme established in two Greek Municipalities for the production of lignocellulosic ethanol using dehydrated household biowaste as a substrate, are presented within this research. This is the first time that biowaste drying was tested at a decentralized level for the production of ethanol using the Simultaneous Saccharification and Fermentation (SSF) process, at a pilot scale in Greece. The decentralized biowaste drying method proved that the household biowaste mass and volume reduction may reach 80% through the dehydration process used. The chemical characteristics related to lignocellulosic ethanol production have proved to differ substantially between seasons thus; special attention should be given to the process applied for ethanol production mainly regarding the enzyme quality and quantity used during the pretreatment stage. The maximum ethanol production achieved was 29.12g/L, approximately 60% of the maximum theoretical yield based on the substrate's sugar content. The use of the decentralized waste drying as an alternative approach for household biowaste minimization and the production of second generation ethanol is considered to be a promising approach for efficient biowaste management and treatment in the future. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Analysis and design of robust decentralized controllers for nonlinear systems

    Energy Technology Data Exchange (ETDEWEB)

    Schoenwald, D.A.

    1993-07-01

    Decentralized control strategies for nonlinear systems are achieved via feedback linearization techniques. New results on optimization and parameter robustness of non-linear systems are also developed. In addition, parametric uncertainty in large-scale systems is handled by sensitivity analysis and optimal control methods in a completely decentralized framework. This idea is applied to alleviate uncertainty in friction parameters for the gimbal joints on Space Station Freedom. As an example of decentralized nonlinear control, singular perturbation methods and distributed vibration damping are merged into a control strategy for a two-link flexible manipulator.

  8. Responsiveness and flexibility in a Decentralized Supply Chain

    DEFF Research Database (Denmark)

    Petersen, Kristian Rasmus; Bilberg, Arne; Hadar, Ronen

    Today’s supply chains are not capable of managing the instabilities that is the case in the market. Instead, there is a need to develop supply chains that are capable of adapting to changes. Through a case study of LEGO, the authors suggest a possible solution: a decentralized supply chain serving...... independent and self-sufficient local factories. The decentralized supply chain is provided with materials, parts and pre-assembled elements from local suppliers and supplies the local market in return. Keywords: Decentralize, Responsiveness, Flexibility...

  9. Dissecting interdomain communication within cAPK regulatory subunit type IIβ using enhanced amide hydrogen/deuterium exchange mass spectrometry (DXMS)

    Science.gov (United States)

    Zawadzki, Kerri M.; Hamuro, Yoshitomo; Kim, Jack S.; Garrod, Siv; Stranz, David D.; Taylor, Susan S.; Woods, Virgil L.

    2003-01-01

    cAMP-dependent protein kinase (cAPK) is a heterotetramer containing a regulatory (R) subunit dimer bound to two catalytic (C) subunits and is involved in numerous cell signaling pathways. The C-subunit is activated allosterically when two cAMP molecules bind sequentially to the cAMP-binding domains, designated A and B (cAB-A and cAB-B, respectively). Each cAMP-binding domain contains a conserved Arg residue that is critical for high-affinity cAMP binding. Replacement of this Arg with Lys affects cAMP affinity, the structural integrity of the cAMP-binding domains, and cAPK activation. To better understand the local and long-range effects that the Arg-to-Lys mutation has on the dynamic properties of the R-subunit, the amide hydrogen/deuterium exchange in the RIIβ subunit was probed by electrospray mass spectrometry. Mutant proteins containing the Arg-to-Lys substitution in either cAMP-binding domain were deuterated for various times and then, prior to mass spectrometry analysis, subjected to pepsin digestion to localize the deuterium incorporation. Mutation of this Arg in cAB-A (Arg230) causes an increase in amide hydrogen exchange throughout the mutated domain that is beyond the modest and localized effects of cAMP removal and is indicative of the importance of this Arg in domain organization. Mutation of Arg359 (cAB-B) leads to increased exchange in the adjacent cAB-A domain, particularly in the cAB-A domain C-helix that lies on top of the cAB-B domain and is believed to be functionally linked to the cAB-B domain. This interdomain communication appears to be a unidirectional pathway, as mutation of Arg230 in cAB-A does not effect dynamics of the cAB-B domain. PMID:12930997

  10. Changes in water mass exchange between the NW shelf areas and the North Atlantic and their impact on nutrient/carbon cycling

    Science.gov (United States)

    Gröger, Matthias; Maier-Reimer, Ernst; Mikolajewicz, Uwe; Segschneider, Joachim; Sein, Dimitry

    2010-05-01

    Despite their comparatively small extension on a global scale, shelf areas are of interest for several economic reasons and climatic processes related to nutrient cycling, sea food supply, and biological productivity. Moreover, they constitute an important interface for nutrients, pollutants and freshwater on their pathway from the continents to the open ocean. This modelling study aims to investigate the spatial and temporal variability of water mass exchange between the North Atlantic and the NW European shelf and their impact on nutrient/carbon cycling and biological productivity. For this, a new modeling approach has been set up which bridges the gap between pure shelf models where water mass transports across the model domain too strongly depend on the formulation of open boundaries and global models suffering under their too coarse resolution in shelf regions. The new model consists of the global ocean and carbon cycle model MPIOM/HAMOCC with strongly increased resolution in the North Sea and the North Atlantic coupled to the regional atmosphere model REMO. The model takes the full luni-solar tides into account. It includes further a 12 layer sediment module with the relevant pore water chemistry. The main focus lies on the governing mechanisms of water mass exchange across the shelf break and the imprint on shelf biogeochemistry. For this, artificial tracers with a prescribed decay rate have been implemented to distinguish waters arriving from polar and shelf regions and those that originate from the tropics. Experiments were carried out for the years 1948 - 2007. The relationship to larger scale circulation patterns like the position and variability of the subtropical and subpolar gyres is analyzed. The water mass exchange is analyzed with respect to the nutrient concentration and productivity on the European shelf areas. The implementation of tides leads to an enhanced vertical mixing which causes lower sea surface temperatures compared to simulations

  11. Measuring dynamics in weakly structured regions of proteins using microfluidics-enabled subsecond H/D exchange mass spectrometry.

    Science.gov (United States)

    Rob, Tamanna; Liuni, Peter; Gill, Preet Kamal; Zhu, Shaolong; Balachandran, Naresh; Berti, Paul J; Wilson, Derek J

    2012-04-17

    This work introduces an integrated microfluidic device for measuring rapid H/D exchange (HDX) in proteins. By monitoring backbone amide HDX on the millisecond to low second time scale, we are able to characterize conformational dynamics in weakly structured regions, such as loops and molten globule-like domains that are inaccessible in conventional HDX experiments. The device accommodates the entire MS-based HDX workflow on a single chip with residence times sufficiently small (ca. 8 s) that back-exchange is negligible (≤5%), even without cooling. Components include an adjustable position capillary mixer providing a variable-time labeling pulse, a static mixer for HDX quenching, a proteolytic microreactor for rapid protein digestion, and on-chip electrospray ionization (ESI). In the present work, we characterize device performance using three model systems, each illustrating a different application of 'time-resolved' HDX. Ubiquitin is used to illustrate a crude, high throughput structural analysis based on a single subsecond HDX time-point. In experiments using cytochrome c, we distinguish dynamic behavior in loops, establishing a link between flexibility and interactions with the heme prosthetic group. Finally, we localize an unusually high 'burst-phase' of HDX in the large tetrameric enzyme DAHP synthase to a 'molten globule-like' region surrounding the active site.

  12. Measuring the hydrogen/deuterium exchange of proteins at high spatial resolution by mass spectrometry: overcoming gas-phase hydrogen/deuterium scrambling.

    Science.gov (United States)

    Rand, Kasper D; Zehl, Martin; Jørgensen, Thomas J D

    2014-10-21

    Proteins are dynamic molecules that exhibit conformational flexibility to function properly. Well-known examples of this are allosteric regulation of protein activity and ligand-induced conformational changes in protein receptors. Detailed knowledge of the conformational properties of proteins is therefore pertinent to both basic and applied research, including drug development, since the majority of drugs target protein receptors and a growing number of drugs introduced to the market are therapeutic peptides or proteins. X-ray crystallography provides a static picture at atomic resolution of the lowest-energy structure of the native ensemble. There is a growing need for sensitive analytical tools to explore all of the significant molecular structures in the conformational landscape of proteins. Hydrogen/deuterium exchange monitored by mass spectrometry (HDX-MS) has recently emerged as a powerful method for characterizing protein conformational dynamics. The basis of this method is the fact that backbone amides in stable hydrogen-bonded structures (e.g., α-helices and β-sheets) are protected against exchange with the aqueous solvent. All protein structures are dynamic, however, and eventually all of the protecting hydrogen bonds will transiently break as the protein--according to thermodynamic principles--cycles through partially unfolded states that correspond to excited free energy levels. As a result, all of the backbone amides will eventually become temporarily solvent-exposed and exchange-competent over time. Consequently, a folded protein in D2O will gradually incorporate deuterium into its backbone amides, and the kinetics of the process can be readily monitored by mass spectrometry. The deuterium uptake kinetics for the intact protein (global exchange kinetics) represents the sum of the exchange kinetics for the individual backbone amides. Local exchange kinetics is typically achieved by using pepsin digestion under quench conditions (i.e., under cold

  13. Recombinant immobilized rhizopuspepsin as a new tool for protein digestion in hydrogen/deuterium exchange mass spectrometry

    Czech Academy of Sciences Publication Activity Database

    Rey, M.; Man, Petr; Brandolin, G.; Forest, E.; Pelosi, L.

    2009-01-01

    Roč. 23, č. 21 (2009), s. 3431-3438 ISSN 0951-4198 Institutional research plan: CEZ:AV0Z50200510 Keywords : mass spectrometry * recombinant immobilized rhizopuspepsin * protease Subject RIV: CE - Biochemistry Impact factor: 2.695, year: 2009

  14. An interactive heuristic for financial planning in decentralized organizations.

    NARCIS (Netherlands)

    M.H. Goedhart; J. Spronk (Jaap)

    1995-01-01

    textabstractPlanning and controlling overall performance in decentralized organizations is a complex task for central management because it is contronted with incomplete information on organizational opportunities and possible conflicts of interest with local managers. Many formal procedures for the

  15. Decentralized Detection in Censoring Sensor Networks under Correlated Observations

    Directory of Open Access Journals (Sweden)

    Abdullah S. Abu-Romeh

    2010-01-01

    Full Text Available The majority of optimal rules derived for different decentralized detection application scenarios are based on an assumption that the sensors' observations are statistically independent. Deriving the optimal decision rule in the canonical decentralized setting with correlated observations was shown to be complicated even for the simple case of two sensors. We introduce an alternative suboptimal rule to deal with correlated observations in decentralized detection with censoring sensors using a modified generalized likelihood ratio test (mGLRT. In the censoring scheme, sensors either send or do not send their complete observations to the fusion center. Using ML estimation to estimate the censored values, the decentralized problem is converted to a centralized problem. Our simulation results indicate that, when sensor observations are correlated, the mGLRT gives considerably better performance in terms of probability of detection than does the optimal decision rule derived for uncorrelated observations.

  16. 8-8-08 International Conference on Decentralization, local power ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    hallas

    2008-08-08

    Saharan Africa, South Asia, and. Latin America. The research findings represent the first regional and global comparative analyses of women's rights, citizenship and governance under decentralization systems and processes.

  17. Engaging Social Capital for Decentralized Urban Stormwater Management

    Science.gov (United States)

    Decentralized approaches to urban stormwater management, whereby installations of green infrastructure (e.g., rain gardens, bioswales, and constructed wetlands) are dispersed throughout a management area, are cost-effective solutions with co-benefits beyond water abatement. Inste...

  18. Papers by the Decentralized Wastewater Management MOU Partnership

    Science.gov (United States)

    Four position papers for state, local, and tribal government officials and interested stakeholders. These papers include information on the uses and benefits of decentralized wastewater treatment and examples of its effective use.

  19. Local Experiences in Decentralization in West and Central Africa ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    associations, private sector) has an essential role to play in the implementation of development activities that fall to local communities. But to what extent do current processes of decentralization allow the integration of actors favorable to ...

  20. DECENTRALIZATION IN THE SYSTEM OF NATIONAL ECONOMY MANAGEMENT

    Directory of Open Access Journals (Sweden)

    Stepaniuk Nataliia

    2018-03-01

    Full Text Available Introduction. Article deals with the investigation of the theoretical approaches to the notion of decentralization in the system of management of the national economy. Purpose. It has been found that for the effective functioning of the state it is necessary to achieve a rational relationship between centralization and decentralization, change the role, responsibility and powers for local self-government and executive authority. Results. t is substantiated that most of the scientific works are devoted to the study of the issue of decentralization of power, the implementation of reform of public finances, the transfer of power to the place as a guarantee of the development of the national economy. It is emphasized that the main idea of decentralization is to transfer competence to local government to address local needs issues. Consequently, decentralization is closely linked to the organization of public administration, promotes the building of effective relations between state authorities and local government. The main advantages of decentralization are: simplified management on the local area, establishing closer connection with civil society, increasing transparency of managerial decisions and raising the level of responsibility to the territorial community. Considered organizational and legal aspects of introduction of decentralization in Ukraine. It is noted that the course on decentralization outlines both prospects and implementation problems. Among the main risks of decentralization are the inconsistencies of the development of separate territorial units and strategic goals, the loss of state mobility, reduction of workplaces of the state apparatus, risks of complication of coordination between levels of management. Conclusions. It has been determined that for efficiency and effectiveness of the reform decentralization principles are necessary for wide introduction in the administrative, political, budgetary, financial and social spheres

  1. Corruption and government spending : The role of decentralization

    OpenAIRE

    Korneliussen, Kristine

    2009-01-01

    This thesis points to a possible weakness of the empirical literature on corruption and government spending. That corruption affects the composition of government spending, and in particular that it affects education and health spending adversely, seems to be empirically well established. However, there exist additional literature closely related to corruption and government spending, treating(i) a relationship between corruption and decentralization, and (ii) a relationship between decentral...

  2. Decentralized Investment Management: Evidence from the Pension Fund Industry

    OpenAIRE

    Blake, David; Timmermann, Allan G; Tonks, Ian; Wermers, Russ

    2010-01-01

    The past few decades have seen amajor shift from centralized to decentralized investment management by pension fund sponsors, despite the increased coordination problems that this brings. Using a unique, proprietary dataset of pension sponsors and managers, we identify two secular decentralization trends: sponsors switched (i) from generalist (balanced) to specialist managers across asset classes and (ii) from single to multiple competing managers within each asset class. We study the effect ...

  3. Corruption, accountability, and decentralization: theory and evidence from Mexico

    OpenAIRE

    Goodspeed, Timothy J.

    2011-01-01

    One of the fundamental tenets of fiscal federalism is that, absent various sorts of externalities, decentralized governments that rely on own-source revenues should be more fiscally efficient than decentralized governments that rely on grant financing. The argument relies in part on the idea that sub-national governments, being closer to the people, are more accountable to its citizens. Accountability to citizens is also important in understanding the presence of corruption in government. Thi...

  4. Emergent Semantics Interoperability in Large-Scale Decentralized Information Systems

    CERN Document Server

    Cudré-Mauroux, Philippe

    2008-01-01

    Peer-to-peer systems are evolving with new information-system architectures, leading to the idea that the principles of decentralization and self-organization will offer new approaches in informatics, especially for systems that scale with the number of users or for which central authorities do not prevail. This book describes a new way of building global agreements (semantic interoperability) based only on decentralized, self-organizing interactions.

  5. Computational State Transfer: An Architectural Style for Decentralized Systems

    OpenAIRE

    Gorlick, Michael Martin

    2016-01-01

    A decentralized system is a distributed system that operates under multiple, distinct spheres of authority in which collaboration among the principals is characterized by mutual distrust. Now commonplace, decentralized systems appear in a number of disparate domains: commerce, logistics, medicine, software development, manufacturing, and financial trading to name but a few. These systems of systems face two overlapping demands: security and safety to protect against errors, omissions and thre...

  6. Subsidiarity in Principle: Decentralization of Water Resources Management

    Directory of Open Access Journals (Sweden)

    Ryan Stoa

    2014-05-01

    Full Text Available The subsidiarity principle of water resources management suggests that water management and service delivery should take place at the lowest appropriate governance level. The principle is attractive for several reasons, primarily because: 1 the governance level can be reduced to reflect environmental characteristics, such as the hydrological borders of a watershed that would otherwise cross administrative boundaries; 2 decentralization promotes community and stakeholder engagement when decision-making is localized; 3 inefficiencies are reduced by eliminating reliance on central government bureaucracies and budgetary constraints; and 4 laws and institutions can be adapted to reflect localized conditions at a scale where integrated natural resources management and climate change adaptation is more focused. Accordingly, the principle of subsidiarity has been welcomed by many states committed to decentralized governance, integrated water resources management, and/or civic participation. However, applications of decentralization have not been uniform, and in some cases have produced frustrating outcomes for states and water resources. Successful decentralization strategies are heavily dependent on dedicated financial resources and human resource capacity. This article explores the nexus between the principle of subsidiarity and the enabling environment, in the hope of articulating factors likely to contribute to, or detract from, the success of decentralized water resources management. Case studies from Haiti, Rwanda, and the United States’ Florida Water Management Districts provide examples of the varied stages of decentralization.

  7. Rethinking Decentralization in Education in terms of Administrative Problems

    Directory of Open Access Journals (Sweden)

    Vasiliki Papadopoulou

    2013-11-01

    Full Text Available The general purpose of this study is to thoroughly examine decentralization in education according to the literature and previous research, and to discuss the applicability of educational decentralization practices in Turkey. The literature was reviewed for the study and findings reported. It has been observed that decentralization in education practices were realized in many countries after the 1980’s. It is obvious that the educational system in Turkey has difficulty in meeting the needs, and encounters many problems due to its present centralist state. Educational decentralization can provide effective solutions for stakeholder engagement, educational financing and for problems in decision making and operation within the education system. However, the present state of local governments, the legal framework, geographical, cultural and social features indicate that Turkey’s conditions are not ready for decentralization in education. A decentralization model realized in the long run according to Turkey’s conditions, and as a result of a social consensus, can help resolve the problems of the Turkish education system.

  8. FISCAL DECENTRALIZATION IN THE DRC: EVIDENCE OFREVENUE ASSIGNMENT

    Directory of Open Access Journals (Sweden)

    Angelita Kithatu-Kiwekete

    2017-07-01

    Full Text Available The rationalefor central government to devolve resources for service provisionhas been debated in decentralization literature. Decentralization enhancesdemocracy,encouragesparticipation in local development initiativesandpromotes local political accountability.This discourse has been complemented bythe implementation of fiscal decentralization to increase the ability of sub-nationalgovernment in financing municipal service delivery. Fiscal decentralization hasoften been adopted by African statessince the onset ofthe New PublicManagement erain an effortto improvethe standard ofgovernance. The concernis that African states have taken minimal steps to adopt fiscal devolution thatpromotes revenue assignment which in turn limits sub-nationalgovernments’ability to generate own source revenues.This article examines the revenue assignment function of fiscal decentralization inthe Democratic Republic of Congo(DRCinthelight of decentralizationconcerns that have been raised by civil society, as the country charts its course todemocracy. The article is a desktop study that will consider documents andpoliciesin theDRCon thenational, provincialand locallevel as far asstaterevenue sourcesare concerned. Revenue assignment should enable DRC’sprovinces and local authoritiestogeneratesignificantrevenueindependently.However, post-conflict reconstruction and development efforts in the Great Lakesregion and in the DRC have largely isolated decentralization which wouldotherwise entrench local fiscalautonomy infinancing for local services anddevelopment. The article concludes that revenue generation for local authoritiesandtheprovinces in the DRC is still very centralised by the national government.Thearticleproposes policy recommendations that will be useful for the country toensurethatdecentralization effortsinclude fiscal devolution toenhance thefinancing for local development initiatives.

  9. FISCAL DECENTRALIZATION IN ALBANIA: EFFECTS OF TERRITORIAL AND ADMINISTRATIVE REFORM

    Directory of Open Access Journals (Sweden)

    Mariola KAPIDANI

    2015-12-01

    Full Text Available The principle of decentralization is a fundamental principle for the establishment and operation of local government. It refers to the process of redistributing the authority and responsibility for certain functions from central government to local government units. In many countries, particularly in developing countries, fiscal decentralization and local governance issues are addressed as highly important to the economic development. According to Stigler (1957, fiscal decentralization brings government closer to the people and a representative government works best when it is closer to the people. Albania is still undergoing the process of decentralization in all aspects: political, economic, fiscal and administrative. Decentralization process is essential to sustainable economic growth and efficient allocation of resources to meet the needs of citizens. Albania has a fragmented system of local government with a very large number of local government units that have neither sufficient fiscal or human capacity to provide public services at a reasonable level (World Bank. However, recent administrative and territorial reform is expected to have a significant impact in many issues related to local autonomy and revenue management. This paper is focused on the progress of fiscal decentralization process in Albania, stating key issues and ongoing challenges for an improved system. The purpose of this study is to analyze the effects of recent territorial reform, identifying problems and opportunities to be addressed in the future.

  10. One dimensional transient numerical study of the mass heat and charge transfer in a proton exchange membrane for PEMFC

    Energy Technology Data Exchange (ETDEWEB)

    Haddad, Djamel; Benmoussa, Hocine [Laboratory (LESEI), Faculty of Engineering, University of Batna (Algeria); Bourmada, Noureddine; Oulmi, Kafia [Laboratory LCCE, Faculty of Science, University of Batna (Algeria); Mahmah, Bouziane; Belhamel, Maiouf [CDER, BP, 62 Avenue-Observatoire, Bouzareah, Alger (Algeria)

    2009-06-15

    The objective of our study is to quantify the mass water transferred by various modes: diffusion, convection and migration. For the water transfer, the principal forces considered in the model are, the convection force, the osmotic force (i.e. diffusion) and the electric force (migration). The first of these forces results from a pressure gradient, the second of a concentration gradient and the third of a protons' migration from the anode to the cathode, which has an effect on the dipole of the water molecules (resistance force to the advancement). The numerical tool used to solve the equations' system is the finite element method. The results obtained numerically considering this method are concentration profiles and concentration variation with time and membrane thickness. These results illustrate the contribution of each mass transfer mode. (author)

  11. Formation of the black-hole binary M33 X-7 through mass exchange in a tight massive system.

    Science.gov (United States)

    Valsecchi, Francesca; Glebbeek, Evert; Farr, Will M; Fragos, Tassos; Willems, Bart; Orosz, Jerome A; Liu, Jifeng; Kalogera, Vassiliki

    2010-11-04

    The X-ray source M33 X-7 in the nearby galaxy Messier 33 is among the most massive X-ray binary stellar systems known, hosting a rapidly spinning, 15.65M(⊙) black hole orbiting an underluminous, 70M(⊙) main-sequence companion in a slightly eccentric 3.45-day orbit (M(⊙), solar mass). Although post-main-sequence mass transfer explains the masses and tight orbit, it leaves unexplained the observed X-ray luminosity, the star's underluminosity, the black hole's spin and the orbital eccentricity. A common envelope phase, or rotational mixing, could explain the orbit, but the former would lead to a merger and the latter to an overluminous companion. A merger would also ensue if mass transfer to the black hole were invoked for its spin-up. Here we report simulations of evolutionary tracks which reveal that if M33 X-7 started as a primary body of 85M(⊙)-99M(⊙) and a secondary body of 28M(⊙)-32M(⊙), in a 2.8-3.1-d orbit, its observed properties can be consistently explained. In this model, the main-sequence primary transfers part of its envelope to the secondary and loses the rest in a wind; it ends its life as a ∼16M(⊙) helium star with an iron-nickel core that collapses to a black hole (with or without an accompanying supernova). The release of binding energy, and possibly collapse asymmetries, 'kick' the nascent black hole into an eccentric orbit. Wind accretion explains the X-ray luminosity, and the black-hole spin can be natal.

  12. On the incidence of Kelvin-Helmholtz instability for mass exchange process at the Earth’s magnetopause

    Directory of Open Access Journals (Sweden)

    R. Smets

    Full Text Available Due to the velocity shear imposed by the solar wind flowing around the magnetosphere, the magnetopause flanks are preferred regions for the development of a Kelvin-Helmholtz instability. Since its efficiency for momentum transfer across the magnetopause has already been established, we investigate its efficiency for mass transfer. Using nonresistive magnetohydrodynamic simulations to describe the magnetic field shape in the instability region, we use test-particle calculations to analyse particle dynamics. We show that the magnetopause thickness and the instability wave-length are too large to lead to nonadiabatic motion of thermal electrons from the magnetosphere. On the other hand, the large mass of H+, He+ and O+ ions leads to such nonadiabatic motion and we thus propose the Kelvin-Helmholtz instability as a mechanism for either magnetospheric ion leakage into the magnetosheath or solar wind ion entry in the magnetosphere. Test-particle calculations are performed in a dimensionless way to discuss the case of each type of ion. The crossing rate is of the order of 10%. This rate is anti-correlated with shear velocity and instability wavelength. It increases with the magnetic shear. The crossing regions at the magnetopause are narrow and localized in the vicinity of the instability wave front. As a Kelvin-Helmholtz instability allows for mass transfer through the magnetopause without any resistivity, we propose it as an alternate process to reconnection for mass transfer through magnetic boundaries.

    Key words. Magnetospheric physics (magnetopause, cusp and boundary layers; MHD waves and instabilities – Space plasma physics (numerical simulation studies

  13. Steroid and Protein Ligand Binding to Cytochrome P450 46A1 as Assessed by Hydrogen – Deuterium Exchange and Mass Spectrometry

    Science.gov (United States)

    Liao, Wei-Li; Dodder, Nathan G.; Mast, Natalia; Pikuleva, Irina A.; Turko, Illarion V.

    2009-01-01

    Cytochrome P450 46A1 (CYP46A1) is a key enzyme responsible for cholesterol elimination from the brain. This P450 can interact with different steroid substrates and protein redox partners. We utilized hydrogen-deuterium (H-D) exchange mass spectrometry for investigating CYP46A1-ligand interactions. First, we tested the applicability of the H-D exchange methodology and assessed the amide proton exchange in substrate-free and cholesterol sulfate-bound P450. The results showed good correspondence to the available crystal structures and prompted investigation of the CYP46A1 interactions with the two steroid substrates cholesterol and 24S-hydroxycholesterol and the protein redox partner adrenodoxin (Adx). Compared to substrate-free P450, four peptides in cholesterol-bound CYP46A1 (65–80, 109–116, 151–164, and 351–361) and eight peptides in 24S-hydroxycholesterol-bound enzyme (50–64, 65–80, 109–116, 117–125, 129–143, 151–164, 260–270, and 364–373) showed altered deuterium incorporation. Most of these peptides constitute the enzyme active site, whereas the 351–361 peptide is from the region putatively interacting with the redox partner Adx. This also defines the proximal (presumably water) channel that opens in CYP46A1 upon substrate binding. Reciprocal studies of Adx binding to substrate-free and cholesterol sulfate-bound CYP46A1 revealed changes in the deuteration of the Adx-binding site 144–150 and 351–361 peptides, active site 225–239 and 301–313 peptides, and in the 265–276 peptide, whose functional role is not yet known. The data obtained provide structural insights into how substrate and redox partner binding are coordinated and linked to the hydration of the enzyme active site. PMID:19317426

  14. Determination of Histidine pKa Values in the Propeptides of Furin and Proprotein Convertase 1/3 Using Histidine Hydrogen-Deuterium Exchange Mass Spectrometry.

    Science.gov (United States)

    Elferich, Johannes; Williamson, Danielle M; David, Larry L; Shinde, Ujwal

    2015-08-04

    Propeptides of proprotein convertases regulate activation of their protease domains by sensing the organellar pH within the secretory pathway. Earlier experimental work highlighted the importance of a conserved histidine residue within the propeptide of a widely studied member, furin. A subsequent evolutionary analysis found an increase in histidine content within propeptides of secreted eukaryotic proteases compared with their prokaryotic orthologs. However, furin activates in the trans-golgi network at a pH of 6.5 while a paralog, proprotein convertase 1/3, activates in secretory vesicles at a pH of 5.5. It is unclear how a conserved histidine can mediate activation at two different pH values. In this manuscript, we measured the pKa values of histidines within the propeptides of furin and proprotein convertase 1/3 using a histidine hydrogen-deuterium exchange mass spectrometry approach. The high density of histidine residues combined with an abundance of basic residues provided challenges for generation of peptide ions with unique histidine residues, which were overcome by employing ETD fragmentation. During this analysis, we found slow hydrogen-deuterium exchange in residues other than histidine at basic pH. Finally, we demonstrate that the pKa of the conserved histidine in proprotein convertase 1/3 is acid-shifted compared with furin and is consistent with its lower pH of activation.

  15. Effect of detergent binding on cytochrome P450 2B4 structure as analyzed by X-ray crystallography and deuterium-exchange mass spectrometry.

    Science.gov (United States)

    Shah, Manish B; Jang, Hyun-Hee; Wilderman, P Ross; Lee, David; Li, Sheng; Zhang, Qinghai; Stout, C David; Halpert, James R

    2016-09-01

    Multiple crystal structures of CYP2B4 have demonstrated the binding of the detergent 5-cyclohexyl-1-pentyl-β-D-maltoside (CYMAL-5) in a peripheral pocket located adjacent to the active site. To explore the consequences of detergent binding, X-ray crystal structures of the peripheral pocket mutant CYP2B4 F202W were solved in the presence of hexaethylene glycol monooctyl ether (C8E6) and CYMAL-5. The structure in the presence of CYMAL-5 illustrated a closed conformation indistinguishable from the previously solved wild-type. In contrast, the F202W structure in the presence of C8E6 revealed a detergent molecule that coordinated the heme-iron and extended to the protein surface through the substrate access channel 2f. Despite the overall structural similarity of these detergent complexes, remarkable differences were observed in the A, A', and H helices, the F-G cassette, the C-D and β4 loop region. Hydrogen-deuterium exchange mass spectrometry (DXMS) was employed to probe these differences and to test the effect of detergents in solution. The presence of either detergent increased the H/D exchange rate across the plastic regions, and the results obtained by DXMS in solution were consistent in general with the relevant structural snapshots. The study provides insight into effect of detergent binding and the interpretation of associated conformational dynamics of CYP2B4. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Ion Mobility Spectrometry-Hydrogen Deuterium Exchange Mass Spectrometry of Anions: Part 3. Estimating Surface Area Exposure by Deuterium Uptake

    Science.gov (United States)

    Khakinejad, Mahdiar; Ghassabi Kondalaji, Samaneh; Donohoe, Gregory C.; Valentine, Stephen J.

    2016-03-01

    Gas-phase hydrogen deuterium exchange (HDX), collision cross section (CCS) measurement, and molecular dynamics simulation (MDS) techniques were utilized to develop and compare three methods for estimating the relative surface area exposure of separate peptide chains within bovine insulin ions. Electrosprayed [M - 3H]3- and [M - 5H]5- insulin ions produced a single conformer type with respective collision cross sections of 528 ± 5 Å2 and 808 ± 2 Å2. [M - 4H]4- ions were comprised of more compact (Ω = 676 ± 3 Å2) and diffuse (i.e., more elongated, Ω = 779 ± 3 Å2) ion conformer types. Ions were subjected to HDX in the drift tube using D2O as the reagent gas. Collision-induced dissociation was used to fragment mobility-selected, isotopically labeled [M - 4H]4- and [M - 5H]5- ions into the protein subchains. Deuterium uptake levels of each chain can be explained by limited inter-chain isotopic scrambling upon collisional activation. Using nominal ion structures from MDS and a hydrogen accessibility model, the deuterium uptake for each chain was correlated to its exposed surface area. In separate experiments, the per-residue deuterium content for the protonated and deprotonated ions of the synthetic peptide KKDDDDDIIKIIK were compared. The differences in deuterium content indicated the regional HDX accessibility for cations versus anions. Using ions of similar conformational type, this comparison highlights the complementary nature of HDX data obtained from positive- and negative-ion analysis.

  17. Enumeration of labile hydrogens in natural organic matter by use of hydrogen/deuterium exchange Fourier transform ion cyclotron resonance mass spectrometry.

    Science.gov (United States)

    Kostyukevich, Yury; Kononikhin, Alexey; Popov, Igor; Kharybin, Oleg; Perminova, Irina; Konstantinov, Andrey; Nikolaev, Eugene

    2013-11-19

    A method to enumerate labile hydrogens in all constituents of molecular ensemble of natural organic matter (NOM) based on our previously developed simple hydrogen/deuterium (H/D) exchange (electrospray ionization (ESI) ion source (Kostyukevich et al. Anal. Chem. 2013, 85, 5330) and ultra-high-resolution Fourier transform ion cyclotron resonance mass spectrometry is presented. The method was applied for analysis of Suwannee River fulvic acid (SRFA), which is an International Humic Substances Society standard, as well as Siberian crude oil; and lignosulfonate. We found that SRFA and lignosulfonate molecules contain 2-5 labile hydrogens, and their number increases with the number of oxygens in the molecule. Also, we observed that compounds of Siberian crude oil ionizing in positive-ESI mode do not have labile hydrogens, while compounds ionizing in negative-ESI mode have one labile hydrogen that detaches during ESI ionization.

  18. Ligand-induced conformational dynamics of the Escherichia coli Na+/H+ antiporter NhaA revealed by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Eisinger, Martin Lorenz; Dörrbaum, Aline Ricarda; Michel, Hartmut; Padan, Etana; Langer, Julian David

    2017-10-31

    Na + /H + antiporters comprise a family of membrane proteins evolutionarily conserved in all kingdoms of life and play an essential role in cellular ion homeostasis. The NhaA crystal structure of Escherichia coli has become the paradigm for this class of secondary active transporters. However, structural data are only available at low pH, where NhaA is inactive. Here, we adapted hydrogen/deuterium-exchange mass spectrometry (HDX-MS) to analyze conformational changes in NhaA upon Li + binding at physiological pH. Our analysis revealed a global conformational change in NhaA with two sets of movements around an immobile binding site. Based on these results, we propose a model for the ion translocation mechanism that explains previously controversial data for this antiporter. Furthermore, these findings contribute to our understanding of related human transporters that have been linked to various diseases. Published under the PNAS license.

  19. Structural elucidation of metabolites of ginkgolic acid in rat liver microsomes by ultra-performance liquid chromatography/electrospray ionization tandem mass spectrometry and hydrogen/deuterium exchange.

    Science.gov (United States)

    Liu, Z H; Chen, J; Yu, L S; Jiang, H D; Yao, T W; Zeng, S

    2009-07-01

    Ginkgolic acids have been shown to possess allergenic as well as genotoxic and cytotoxic properties. The question arises whether the metabolism of ginkgolic acids in the liver could decrease or increase their toxicity. In this study, the in vitro metabolism of ginkgolic acid (15:1, GA), one component of ginkgo acids, was investigated as a model compound in Sprague-Dawley rat liver microsomes. The metabolites were analyzed by ultra-performance liquid chromatography coupled with photodiode array detector/negative-ion electrospray ionization tandem mass spectrometry (UPLC-PDA/ESI-MS/MS) and hydrogen/deuterium (H/D) exchange. The result showed that the benzene ring remained unchanged and the oxidations occurred at the side alkyl chain in rat liver microsomes. At least eight metabolites were found. Among them, six phase I metabolites were tentatively identified. This study might be useful for the investigation of toxicological mechanism of ginkgolic acids. Copyright (c) 2009 John Wiley & Sons, Ltd.

  20. An Asymmetric Runaway Domain Swap Antithrombin Dimer as a Key Intermediate for Polymerization Revealed by Hydrogen/Deuterium-Exchange Mass Spectrometry

    DEFF Research Database (Denmark)

    Trelle, Morten Beck; Pedersen, Shona; Østerlund, Eva Christina

    2017-01-01

    analysis by NMR spectroscopy. Here, we show how hydrogen/deuterium-exchange mass spectrometry (HDX-MS) provides detailed insight into the structural dynamics of each subunit in a polymerization-competent antithrombin dimer. Upon deuteration, this dimer surprisingly yields bimodal isotope distributions...... for the majority of peptides, demonstrating an asymmetric configuration of the two subunits. The data reveal that one subunit is very dynamic, potentially intrinsically disordered, whereas the other is considerably less dynamic. The local subunit-specific deuterium uptake of this polymerization-competent dimer...... strongly supports a β4A-β5A β-hairpin runaway domain swap mechanism for antithrombin polymerization. HDX-MS thus holds exceptional promise as an enabling analytical technique in the efforts toward future pharmacological intervention with protein polymerization and associated diseases....

  1. Ligand binding and conformational dynamics in a flavin-based electron-bifurcating enzyme complex revealed by Hydrogen-Deuterium Exchange Mass Spectrometry.

    Science.gov (United States)

    Demmer, Julius K; Rupprecht, Fiona A; Eisinger, Martin L; Ermler, Ulrich; Langer, Julian D

    2016-12-01

    Flavin-based electron bifurcation (FBEB) is a novel mechanism of energy coupling used by anaerobic microorganisms to optimize their energy metabolism efficiency. The first high-resolution structure of a complete FBEB enzyme complex, the NADH-dependent reduced ferredoxin: NADP + -oxidoreductase (NfnAB) of Thermotoga maritima, was recently solved. However, no experimental evidence for the NADPH-binding site and conformational changes during the FBEB reaction are available. Here we analyzed ligand binding and the conformational dynamics of oxygen-sensitive NfnAB using Hydrogen-Deuterium Exchange Mass-Spectrometry, including a customized anaerobic workflow. We confirmed the NADH and the previously postulated NADPH-binding site. Furthermore, we observed an NfnA-NfnB rearrangement upon NADPH binding which supports the proposed FBEB mechanism. © 2016 Federation of European Biochemical Societies.

  2. Conformational Analysis of Large and Highly Disulfide-Stabilized Proteins by Integrating Online Electrochemical Reduction into an Optimized H/D Exchange Mass Spectrometry Workflow

    DEFF Research Database (Denmark)

    Trabjerg, Esben; Jakobsen, Rasmus U.; Mysling, Simon

    2015-01-01

    Analysis of disulfide-bonded proteins by hydrogen/deuterium exchange mass spectrometry (HDX-MS) requires effective and rapid reduction of disulfide bonds before enzymatic digestion in order to increase sequence coverage. In a conventional HDX-MS workflow, disulfide bonds are reduced chemically...... by addition of a reducing agent to the quench solution (e.g., tris(2-carboxyethyl)phosphine (TCEP)). The chemical reduction, however, is severely limited under quenched conditions due to a narrow time window as well as low pH and temperature. Here, we demonstrate the real-world applicability of integrating...... electrochemical reduction into an online HDX-MS workflow. We have optimized the electrochemical reduction efficiency during HDX-MS analysis of two particularly challenging disulfide stabilized proteins: a therapeutic IgG1-antibody and nerve growth factor-β (NGF). Several different parameters (flow rate...

  3. Platform Architecture for Decentralized Positioning Systems

    Directory of Open Access Journals (Sweden)

    Zakaria Kasmi

    2017-04-01

    Full Text Available A platform architecture for positioning systems is essential for the realization of a flexible localization system, which interacts with other systems and supports various positioning technologies and algorithms. The decentralized processing of a position enables pushing the application-level knowledge into a mobile station and avoids the communication with a central unit such as a server or a base station. In addition, the calculation of the position on low-cost and resource-constrained devices presents a challenge due to the limited computing, storage capacity, as well as power supply. Therefore, we propose a platform architecture that enables the design of a system with the reusability of the components, extensibility (e.g., with other positioning technologies and interoperability. Furthermore, the position is computed on a low-cost device such as a microcontroller, which simultaneously performs additional tasks such as data collecting or preprocessing based on an operating system. The platform architecture is designed, implemented and evaluated on the basis of two positioning systems: a field strength system and a time of arrival-based positioning system.

  4. Decentralized provenance-aware publishing with nanopublications

    Directory of Open Access Journals (Sweden)

    Tobias Kuhn

    2016-08-01

    Full Text Available Publication and archival of scientific results is still commonly considered the responsability of classical publishing companies. Classical forms of publishing, however, which center around printed narrative articles, no longer seem well-suited in the digital age. In particular, there exist currently no efficient, reliable, and agreed-upon methods for publishing scientific datasets, which have become increasingly important for science. In this article, we propose to design scientific data publishing as a web-based bottom-up process, without top-down control of central authorities such as publishing companies. Based on a novel combination of existing concepts and technologies, we present a server network to decentrally store and archive data in the form of nanopublications, an RDF-based format to represent scientific data. We show how this approach allows researchers to publish, retrieve, verify, and recombine datasets of nanopublications in a reliable and trustworthy manner, and we argue that this architecture could be used as a low-level data publication layer to serve the Semantic Web in general. Our evaluation of the current network shows that this system is efficient and reliable.

  5. Predicting protein aggregation during storage in lyophilized solids using solid state amide hydrogen/deuterium exchange with mass spectrometric analysis (ssHDX-MS).

    Science.gov (United States)

    Moorthy, Balakrishnan S; Schultz, Steven G; Kim, Sherry G; Topp, Elizabeth M

    2014-06-02

    Solid state amide hydrogen/deuterium exchange with mass spectrometric analysis (ssHDX-MS) was used to assess the conformation of myoglobin (Mb) in lyophilized formulations, and the results correlated with the extent of aggregation during storage. Mb was colyophilized with sucrose (1:1 or 1:8 w/w), mannitol (1:1 w/w), or NaCl (1:1 w/w) or in the absence of excipients. Immediately after lyophilization, samples of each formulation were analyzed by ssHDX-MS and Fourier transform infrared spectroscopy (FTIR) to assess Mb conformation, and by dynamic light scattering (DLS) and size exclusion chromatography (SEC) to determine the extent of aggregation. The remaining samples were then placed on stability at 25 °C and 60% RH or 40 °C and 75% RH for up to 1 year, withdrawn at intervals, and analyzed for aggregate content by SEC and DLS. In ssHDX-MS of samples immediately after lyophilization (t = 0), Mb was less deuterated in solids containing sucrose (1:1 and 1:8 w/w) than in those containing mannitol (1:1 w/w), NaCl (1:1 w/w), or Mb alone. Deuterium uptake kinetics and peptide mass envelopes also indicated greater Mb structural perturbation in mannitol, NaCl, or Mb-alone samples at t = 0. The extent of deuterium incorporation and kinetic parameters related to rapidly and slowly exchanging amide pools (Nfast, Nslow), measured at t = 0, were highly correlated with the extent of aggregation on storage as measured by SEC. In contrast, the extent of aggregation was weakly correlated with FTIR band intensity and peak position measured at t = 0. The results support the use of ssHDX-MS as a formulation screening tool in developing lyophilized protein drug products.

  6. Structure-dependent degradation of polar compounds in weathered oils observed by atmospheric pressure photo-ionization hydrogen/deuterium exchange ultrahigh resolution mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Islam, Ananna; Kim, Donghwi [Kyungpook National University, Department of Chemistry, Daegu 702-701 (Korea, Republic of); Yim, Un Hyuk; Shim, Won Joon [Oil and POPs Research Group, Korea Institute of Ocean Science and Technology, KIOST, Geoje 656-834 (Korea, Republic of); Kim, Sunghwan, E-mail: sunghwank@knu.ac.kr [Kyungpook National University, Department of Chemistry, Daegu 702-701 (Korea, Republic of); Green Nano Center, Department of Chemistry, Daegu 702-701 (Korea, Republic of)

    2015-10-15

    Highlights: • We examined source crude oil and weathered oils from M/V Hebei accident. • APPI hydrogen/deuterium exchange ultrahigh mass spectrometry was applied. • N{sub 1} class compounds with 2° and/or 3° amine decrease in larger scale than pyridines. • Preferential degradation of nitrogen-containing compounds was confirmed. • Significant increase in S{sub 1}O{sub 1} compounds was observed as the weathering proceeds. - Abstract: The resin fractions of fresh mixtures of three oils spilled during the M/V Hebei Spirit oil spill, as well as weathered oils collected at weathering stages II and IV from the oil spill site were analyzed and compared by atmospheric pressure photo-ionization hydrogen/deuterium exchange mass spectrometry (HDX MS). The significantly decreased abundance of N{sup +}· and [N − H + D]{sup +} ions suggested that secondary and tertiary amine-containing compounds were preferentially degraded during the early stage of weathering. [N + H]{sup +} and [N + D]{sup +} ions previously attributed to pyridine-type compounds degraded more slowly than secondary and tertiary amine-containing compounds. The preferential degradation of nitrogen-containing compounds was confirmed by photo-degradation experiments using 15 standard compounds. In addition, significant increases of [S{sub 1}O{sub 1} + H]{sup +} and [S{sub 1}O{sub 1} + D]{sup +} ions with higher DBE values were observed from fresh oil mixtures as compared to stages II and IV samples, and that could be linked with the decrease of higher DBE compounds of the S{sub 1} class. This study presented convincing arguments and evidence demonstrating that secondary and tertiary amines were more vulnerable to photo-degradation than compounds containing pyridine, and hence, preferential degradation depending on chemical structures must be considered in the production of hazardous or toxic components.

  7. Tandem mass spectrometry and hydrogen/deuterium exchange studies of protonated species of 1,1'-bis(diphenylphosphino)-ferrocene oxidative impurity generated during a Heck reaction.

    Science.gov (United States)

    Wu, Lianming; Hernandez-Soto, Heriberto; Liu, David Q; Vogt, Frederick G; O'Neill-Slawecki, Stacy A; Su, Qiaogong

    2008-01-01

    Oxidation of 1,1'-bis(diphenylphosphino)-ferrocene (DPPF) was found to occur when it served as the ligand for Pd(II)(CH3COO)2 in a Heck reaction. This oxidative impurity of DPPF, referred to as DPPF(O), was identified by high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) and exact mass measurements. Protonated DPPF(O) exhibited unique fragmentation pathways in the gas phase. Hydrogen/deuterium (H/D) exchange experiments provided important insights into the dissociation mechanisms of protonated DPPF(O), suggesting the existence of isomeric structures of the product ions by retaining or losing a proton (or deuteron) upon collision-induced dissociation (CID). The specific fate of the proton (or deuteron) upon CID is postulated to be dependent on the distance between the exchangeable proton (or deuteron) and the sites of bond cleavage. Density functional theory (DFT) calculations at the B3LYP/LANL2DZ level of theory showed that oxygen in DPPF(O) plays a pivotal role in invoking pi-cation interactions between the p-type lone pair electrons (n pi) in oxygen and the anti-bonding orbital of Fe(II), accounting for the major fragmentation pathways of protonated DPPF(O). Facile formation of organometallic distonic ions in dissociation of protonated DPPF(O), and especially of protonated DPPF, could be useful for further exploration of their chemical properties by gas-phase ion/molecule reactions. Copyright (c) 2008 John Wiley & Sons, Ltd.

  8. Structure-dependent degradation of polar compounds in weathered oils observed by atmospheric pressure photo-ionization hydrogen/deuterium exchange ultrahigh resolution mass spectrometry

    International Nuclear Information System (INIS)

    Islam, Ananna; Kim, Donghwi; Yim, Un Hyuk; Shim, Won Joon; Kim, Sunghwan

    2015-01-01

    Highlights: • We examined source crude oil and weathered oils from M/V Hebei accident. • APPI hydrogen/deuterium exchange ultrahigh mass spectrometry was applied. • N 1 class compounds with 2° and/or 3° amine decrease in larger scale than pyridines. • Preferential degradation of nitrogen-containing compounds was confirmed. • Significant increase in S 1 O 1 compounds was observed as the weathering proceeds. - Abstract: The resin fractions of fresh mixtures of three oils spilled during the M/V Hebei Spirit oil spill, as well as weathered oils collected at weathering stages II and IV from the oil spill site were analyzed and compared by atmospheric pressure photo-ionization hydrogen/deuterium exchange mass spectrometry (HDX MS). The significantly decreased abundance of N + · and [N − H + D] + ions suggested that secondary and tertiary amine-containing compounds were preferentially degraded during the early stage of weathering. [N + H] + and [N + D] + ions previously attributed to pyridine-type compounds degraded more slowly than secondary and tertiary amine-containing compounds. The preferential degradation of nitrogen-containing compounds was confirmed by photo-degradation experiments using 15 standard compounds. In addition, significant increases of [S 1 O 1 + H] + and [S 1 O 1 + D] + ions with higher DBE values were observed from fresh oil mixtures as compared to stages II and IV samples, and that could be linked with the decrease of higher DBE compounds of the S 1 class. This study presented convincing arguments and evidence demonstrating that secondary and tertiary amines were more vulnerable to photo-degradation than compounds containing pyridine, and hence, preferential degradation depending on chemical structures must be considered in the production of hazardous or toxic components

  9. Improving Patient Schedules by Multi-agent Pareto Appointment Exchanging

    NARCIS (Netherlands)

    I.B. Vermeulen (Ivan); S.M. Bohte (Sander); D.J.A. Somefun (Koye); J.A. La Poutré (Han)

    2006-01-01

    textabstractWe present a dynamic and distributed approach to the hospital patient scheduling problem: the multi-agent Pareto-improvement appointment exchanging algorithm, MPAEX. It respects the decentralization of scheduling authorities and is capable of continuously adjusting the different patient

  10. Analysis of the local dynamics of human insulin and a rapid-acting insulin analog by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Nakazawa, Shiori; Ahn, Joomi; Hashii, Noritaka; Hirose, Kenji; Kawasaki, Nana

    2013-06-01

    Human insulin and insulin lispro (lispro), a rapid-acting insulin analog, have identical primary structures, except for the transposition of a pair of amino acids. This mutation results in alterations in their higher order structures, with lispro dissociating more easily than human insulin. In our previous study performed using hydrogen/deuterium exchange mass spectrometry (HDX/MS), differences were observed in the rates and levels of deuteration among insulin analog products, which were found to be related to their self-association stability. In this study, we carried out peptide mapping of deuterated human insulin and lispro to determine the regions responsible for these deuteration differences and to elucidate the type of structural changes that affect their HDX reactivity. We identified A3-6 and B22-24 as the 2 regions that showed distinct differences in the number of deuterium atoms incorporated between human insulin and lispro. These regions contain residues that are thought to participate in hexamerization and dimerization, respectively. We also determined that over time, the differences in deuteration levels decreased in A3-6, whereas they increased in B22-24, suggesting a difference in the dynamics between these 2 regions. This article is part of a Special Issue entitled: Mass spectrometry in structural biology. Copyright © 2012 Elsevier B.V. All rights reserved.

  11. Mapping the contact surfaces in the Lamin A:AIMP3 complex by hydrogen/deuterium exchange FT-ICR mass spectrometry.

    Directory of Open Access Journals (Sweden)

    Yeqing Tao

    Full Text Available Aminoacyl-tRNA synthetases-interacting multifunctional protein3 (AIMP3/p18 is involved in the macromolecular tRNA synthetase complex via its interaction with several aminoacyl-tRNA synthetases. Recent reports reveal a novel function of AIMP3 as a tumor suppressor by accelerating cellular senescence and causing defects in nuclear morphology. AIMP3 specifically mediates degradation of mature Lamin A (LmnA, a major component of the nuclear envelope matrix; however, the mechanism of how AIMP3 interacts with LmnA is unclear. Here we report solution-phase hydrogen/deuterium exchange (HDX for AIMP3, LmnA, and AIMP3 in association with the LmnA C-terminus. Reversed-phase LC coupled with LTQ 14.5 T Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS results in high mass accuracy and resolving power for comparing the D-uptake profiles for AIMP3, LmnA, and their complex. The results show that the AIMP3-LmnA interaction involves one of the two putative binding sites and an adjacent novel interface on AIMP3. LmnA binds AIMP3 via its extreme C-terminus. Together these findings provide a structural insight for understanding the interaction between AIMP3 and LmnA in AIMP3 degradation.

  12. Mapping the contact surfaces in the Lamin A:AIMP3 complex by hydrogen/deuterium exchange FT-ICR mass spectrometry.

    Science.gov (United States)

    Tao, Yeqing; Fang, Pengfei; Kim, Sunghoon; Guo, Min; Young, Nicolas L; Marshall, Alan G

    2017-01-01

    Aminoacyl-tRNA synthetases-interacting multifunctional protein3 (AIMP3/p18) is involved in the macromolecular tRNA synthetase complex via its interaction with several aminoacyl-tRNA synthetases. Recent reports reveal a novel function of AIMP3 as a tumor suppressor by accelerating cellular senescence and causing defects in nuclear morphology. AIMP3 specifically mediates degradation of mature Lamin A (LmnA), a major component of the nuclear envelope matrix; however, the mechanism of how AIMP3 interacts with LmnA is unclear. Here we report solution-phase hydrogen/deuterium exchange (HDX) for AIMP3, LmnA, and AIMP3 in association with the LmnA C-terminus. Reversed-phase LC coupled with LTQ 14.5 T Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) results in high mass accuracy and resolving power for comparing the D-uptake profiles for AIMP3, LmnA, and their complex. The results show that the AIMP3-LmnA interaction involves one of the two putative binding sites and an adjacent novel interface on AIMP3. LmnA binds AIMP3 via its extreme C-terminus. Together these findings provide a structural insight for understanding the interaction between AIMP3 and LmnA in AIMP3 degradation.

  13. Use of deuterium labeling by high-temperature solid-state hydrogen-exchange reaction for mass spectrometric analysis of bradykinin biotransformation.

    Science.gov (United States)

    Kopylov, Arthur T; Myasoedov, Nikolay F; Dadayan, Alexander K; Zgoda, Victor G; Medvedev, Alexei E; Zolotarev, Yurii A

    2016-06-15

    Studies of molecular biodegradation by mass spectrometry often require synthetic compounds labeled with stable isotopes as internal standards. However, labeling is very expensive especially when a large number of compounds are needed for analysis of biotransformation. Here we describe an approach for qualitative and quantitative analysis using bradykinin (BK) and its in vitro degradation metabolites as an example. Its novelty lies in the use of deuterated peptides which are obtained by a high-temperature solid-state exchange (HSCIE) reaction. Deuterated and native BK were analyzed by positive electrospray ionization high-resolution mass spectrometry (ESI-HRMS) using an Orbitrap Fusion mass spectrometer. High-energy collision-induced dissociation (HCD) experiments were performed on [M+H](+) and [M+2H](2+) ions in targeted-MS(2) mode with adjusted normalized HCD value. After the HSCIE reaction, each amino acid residue of the deuterated peptide contained deuterium atoms and the average degree of substitution was 5.5 atoms per the peptide molecule. The deuterated peptide demonstrated the same chromatographic mobility as the unlabeled counterpart, and lack of racemization during substitution with deuterium. Deuterium-labeled and unlabeled BKs were incubated with human plasma and their corresponding fragments BK(1-5) and BK(1-7), well known as the major metabolites, were detected. Quantitative assays demonstrated applicability of the heavy peptide for both sequencing and quantification of generated fragments. Applicability of the HSCIE deuterated peptide for analysis of routes of its degradation has been shown in in vitro experiments. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  14. Hydrogen/deuterium exchange mass spectrometry reveals specific changes in the local flexibility of plasminogen activator inhibitor 1 upon binding to the somatomedin B domain of vitronectin.

    Science.gov (United States)

    Trelle, Morten Beck; Hirschberg, Daniel; Jansson, Anna; Ploug, Michael; Roepstorff, Peter; Andreasen, Peter A; Jørgensen, Thomas J D

    2012-10-16

    The native fold of plasminogen activator inhibitor 1 (PAI-1) represents an active metastable conformation that spontaneously converts to an inactive latent form. Binding of the somatomedin B domain (SMB) of the endogenous cofactor vitronectin to PAI-1 delays the transition to the latent state and increases the thermal stability of the protein dramatically. We have used hydrogen/deuterium exchange mass spectrometry to assess the inherent structural flexibility of PAI-1 and to monitor the changes induced by SMB binding. Our data show that the PAI-1 core consisting of β-sheet B is rather protected against exchange with the solvent, while the remainder of the molecule is more dynamic. SMB binding causes a pronounced and widespread stabilization of PAI-1 that is not confined to the binding interface with SMB. We further explored the local structural flexibility in a mutationally stabilized PAI-1 variant (14-1B) as well as the effect of stabilizing antibody Mab-1 on wild-type PAI-1. The three modes of stabilizing PAI-1 (SMB, Mab-1, and the mutations in 14-1B) all cause a delayed latency transition, and this effect was accompanied by unique signatures on the flexibility of PAI-1. Reduced flexibility in the region around helices B, C, and I was seen in all three cases, which suggests an involvement of this region in mediating structural flexibility necessary for the latency transition. These data therefore add considerable depth to our current understanding of the local structural flexibility in PAI-1 and provide novel indications of regions that may affect the functional stability of PAI-1.

  15. Insights into small heat shock protein and substrate structure during chaperone action derived from hydrogen/deuterium exchange and mass spectrometry.

    Science.gov (United States)

    Cheng, Guilong; Basha, Eman; Wysocki, Vicki H; Vierling, Elizabeth

    2008-09-26

    Small heat shock proteins (sHSPs) and the related alpha-crystallins are ubiquitous chaperones linked to neurodegenerative diseases, myopathies, and cataract. To better define their mechanism of chaperone action, we used hydrogen/deuterium exchange and mass spectrometry (HXMS) to monitor conformational changes during complex formation between the structurally defined sHSPs, pea PsHsp18.1, and wheat TaHsp16.9, and the heat-denatured model substrates malate dehydrogenase (MDH) and firefly luciferase. Remarkably, we found that even when complexed with substrate, the highly dynamic local structure of the sHSPs, especially in the N-terminal arm (>70% exchange in 5 s), remains unchanged. These results, coupled with sHSP-substrate complex stability, indicate that sHSPs do not adopt new secondary structure when binding substrate and suggest sHSPs are tethered to substrate at multiple sites that are locally dynamic, a feature that likely facilitates recognition and refolding of sHSP-bound substrate by the Hsp70/DnaK chaperone system. Both substrates were found to be stabilized in a partially unfolded state that is observed only in the presence of sHSP. Furthermore, peptide-level HXMS showed MDH was substantially protected in two core regions (residues 95-156 and 228-252), which overlap with the MDH structure protected in the GroEL-bound MDH refolding intermediate. Significantly, despite differences in the size and structure of TaHsp16.9-MDH and PsHsp18.1-MDH complexes, peptide-level HXMS patterns for MDH in both complexes are virtually identical, indicating that stabilized MDH thermal unfolding intermediates are not determined by the identity of the sHSP.

  16. Folding kinetics of the S100A11 protein dimer studied by time-resolved electrospray mass spectrometry and pulsed hydrogen-deuterium exchange.

    Science.gov (United States)

    Pan, Jingxi; Rintala-Dempsey, Anne C; Li, Yu; Shaw, Gary S; Konermann, Lars

    2006-03-07

    This study reports the application of electrospray ionization (ESI) mass spectrometry (MS) with on-line rapid mixing for millisecond time-resolved studies of the refolding and assembly of a dimeric protein complex. Acid denaturation of S100A11 disrupts the native homodimeric protein structure. Circular dichroism and HSQC nuclear magnetic resonance measurements reveal that the monomeric subunits unfold to a moderate degree but retain a significant helicity and some tertiary structural elements. Following a rapid change in solution conditions to a slightly basic pH, the native protein reassembles with an effective rate constant of 6 s(-)(1). The ESI charge state distributions measured during the reaction suggest the presence of three kinetic species, namely, a relatively unfolded monomer (M(U)), a more tightly folded monomeric reaction intermediate (M(F)), and dimeric S100A11. These three forms exhibit distinct calcium binding properties, with very low metal loading levels for M(U), up to two calcium ions for M(F), and up to four for the dimer. Surprisingly, on-line pulsed hydrogen-deuterium exchange (HDX) reveals that each of the monomeric forms of the protein comprises two subspecies that can be distinguished on the basis of their isotope exchange levels. As the reaction proceeds, the more extensively labeled species are depleted. The exponential nature of the measured intensity-time profiles implies that the rate-determining step of the overall process is a unimolecular event. The kinetics are consistent with a sequential folding and assembly mechanism involving two increasingly nativelike monomeric intermediates en route to the native S100A11 dimer.

  17. Mechanism of intracellular cAMP sensor Epac2 activation: cAMP-induced conformational changes identified by amide hydrogen/deuterium exchange mass spectrometry (DXMS).

    Science.gov (United States)

    Li, Sheng; Tsalkova, Tamara; White, Mark A; Mei, Fang C; Liu, Tong; Wang, Daphne; Woods, Virgil L; Cheng, Xiaodong

    2011-05-20

    Epac2, a guanine nucleotide exchange factor, regulates a wide variety of intracellular processes in response to second messenger cAMP. In this study, we have used peptide amide hydrogen/deuterium exchange mass spectrometry to probe the solution structural and conformational dynamics of full-length Epac2 in the presence and absence of cAMP. The results support a mechanism in which cAMP-induced Epac2 activation is mediated by a major hinge motion centered on the C terminus of the second cAMP binding domain. This conformational change realigns the regulatory components of Epac2 away from the catalytic core, making the later available for effector binding. Furthermore, the interface between the first and second cAMP binding domains is highly dynamic, providing an explanation of how cAMP gains access to the ligand binding sites that, in the crystal structure, are seen to be mutually occluded by the other cAMP binding domain. Moreover, cAMP also induces conformational changes at the ionic latch/hairpin structure, which is directly involved in RAP1 binding. These results suggest that in addition to relieving the steric hindrance imposed upon the catalytic lobe by the regulatory lobe, cAMP may also be an allosteric modulator directly affecting the interaction between Epac2 and RAP1. Finally, cAMP binding also induces significant conformational changes in the dishevelled/Egl/pleckstrin (DEP) domain, a conserved structural motif that, although missing from the active Epac2 crystal structure, is important for Epac subcellular targeting and in vivo functions. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

  18. Multi-state steric mass action model and case study on complex high loading behavior of mAb on ion exchange tentacle resin.

    Science.gov (United States)

    Diedrich, Juliane; Heymann, William; Leweke, Samuel; Hunt, Stephen; Todd, Robert; Kunert, Christian; Johnson, Will; von Lieres, Eric

    2017-11-24

    Tentacle resins for IEX are increasingly applied in preparative chromatography for their higher selectivity and higher capacities in comparison to IEX resins without tentacles. However, tentacle resins are often observed to cause unusual elution behavior of monoclonal antibodies under high loading conditions. Understanding this elution behavior is important for a quality by design approach, as it is now mandated by regulatory agencies. A model-based analysis of load, wash and gradient elution is performed for a monoclonal antibody (mAb) on Fractogel SO 3 - . Four experiments with increasing loaded mass show complex peak shapes and formation of a shoulder under overloaded conditions. We hypothesize that the observed peak shapes are caused by mAbs binding in multiple states on the tentacle ion-exchange resin. A new multi-state SMA binding model is used for testing this hypothesis. A two-state binding model is found to quantitatively reproduce all four experiments. An in-depth analysis reveals that the shoulder formation under overloaded conditions can be explained by multi-state binding that particularly manifests in rapid but weak re-adsorption of eluting molecules near the column end. The introduced multi-state SMA model combines features of the so-called spreading model (multiple bound states) and of the standard SMA model (salt dependency). It is by no means limited to ion-exchange chromatography on tentacle resins, but the same concept can be applied for studying systems that are based on other physical mechanisms. The new model can potentially improve mechanistic understanding and facilitate quantitative simulation of various phenomena, such as caused by reorientation, reconformation or unfolding of bound species. Similar concepts can be applied for studying surface-induced aggregation and denaturation. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Development and application of out-of-focus imaging in order to characterize heat and mass exchanges in two-phase flows; Comparaison de diagnostics optiques granulometriques: imagerie en defaut de mise au point - anemometrie phase doppler

    Energy Technology Data Exchange (ETDEWEB)

    Lemaitre, P.; Porcheron, E.; Marchand, D.; Nuboer, A.; Bouilloux, L.; Vendel, J

    2007-07-01

    The aim of this paper is to present the capacity of the out-of-focus imaging in order to measure droplets size in presence of heat and mass exchanges. It is supported with optical simulations first based on geometrical optics, and then with the Lorenz-Mie theory. Finally, this technique is applied in presence of heat and mass transfers in the TOSQAN experiment. (authors)

  20. Decentralized Interleaving of Paralleled Dc-Dc Buck Converters: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, Brian B [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Rodriguez, Miguel [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Sinha, Mohit [University of Minnesota; Dhople, Sairaj [University of Minnesota; Poon, Jason [University of California at Berkeley

    2017-09-01

    We present a decentralized control strategy that yields switch interleaving among parallel connected dc-dc buck converters without communication. The proposed method is based on the digital implementation of the dynamics of a nonlinear oscillator circuit as the controller. Each controller is fully decentralized, i.e., it only requires the locally measured output current to synthesize the pulse width modulation (PWM) carrier waveform. By virtue of the intrinsic electrical coupling between converters, the nonlinear oscillator-based controllers converge to an interleaved state with uniform phase-spacing across PWM carriers. To the knowledge of the authors, this work represents the first fully decentralized strategy for switch interleaving of paralleled dc-dc buck converters.

  1. The Bases of Federalism and Decentralization in Education

    Directory of Open Access Journals (Sweden)

    Carlos Ornelas

    2003-05-01

    Full Text Available This essay uses the Weberian-type ideal to define the conceptual bases of federalism and the decentralization of education. Classic federalism, ficticious federalism (corporativism, the origins and the indigenous version of the new federalism are discussed. We conclude that Mexican constitutional federalism is baroque and ambiguous. Based on theory and the experiences of various countries, bureaucratic centralism and its main characteristics are defined. As a contrast, a typology of educational decentralization is developed. Taken into account are its political, judicial and administrative definitions; a distinction is made between delegation and decentralization. It is argued that with the signing of the Agreement for the Modernization of Basic Education, the Mexican government sought to increase its legitimacy without losing control of education.

  2. [Decentralization of epidemiological surveillance in Pernambuco State, Brazil].

    Science.gov (United States)

    Albuquerque, Ana Coelho de; Mota, Eduardo Luiz Andrade; Felisberto, Eronildo

    2015-04-01

    The study aimed to analyze the relationship between decentralization of the Brazilian Unified National Health System (SUS) and the development of epidemiological surveillance activities in municipalities (counties) in Pernambuco State, Brazil. This was an exploratory descriptive qualitative and quantitative study, including a document search, completion of semi-structured interviews by key informants, and an ecological spatial and time trend study of selected health indicators, covering a 10-year period (2001-2010). The study showed that municipalities adhered to the decentralization process, which was making progress in Pernambuco, but with inequalities and weaknesses in its development. There was also a fluctuation in the time series for the selected indicators. Thus, even though the decentralization of epidemiological surveillance is still incipient in some municipalities, their protagonist role in implementing the activities promotes empowerment at the local level by producing key information for decision-making.

  3. A Retrospective Analysis of the Development of Fiscal Decentralization

    Directory of Open Access Journals (Sweden)

    Rekova Nataliia Yu.

    2017-12-01

    Full Text Available The study forms the theoretical basis for the implementation of fiscal decentralization in Ukraine on the basis of determining the correspondence between the evolution of scientific approaches to the formation of an effective model of public administration and the degree of power centralization at a particular stage of the development of society. The views of thinkers of the ancient states of Egypt, Mesopotamia, India, China, Rome, Greece are generalized, and the priority of centralized public administration without segregation of centralization forms is determined. The degree of centralization in the period of development of feudal states is characterized. The scientific views of representatives of the neoinstitutional direction of economic thought are analyzed in detail, and the stages of the formation of decentralization, in particular fiscal, as a separate theory, are defined. The stages of and the corresponding organizational and legislative documents for the implementation of decentralization in Ukraine are outlined, and its results are characterized.

  4. Laser Processed Heat Exchangers

    Science.gov (United States)

    Hansen, Scott

    2017-01-01

    The Laser Processed Heat Exchanger project will investigate the use of laser processed surfaces to reduce mass and volume in liquid/liquid heat exchangers as well as the replacement of the harmful and problematic coatings of the Condensing Heat Exchangers (CHX). For this project, two scale unit test articles will be designed, manufactured, and tested. These two units are a high efficiency liquid/liquid HX and a high reliability CHX.

  5. SecuRES: Secure Resource Sharing System : AN INVESTIGATION INTO USE OF PUBLIC LEDGER TECHNOLOGY TO CREATE DECENTRALIZED DIGITAL RESOURCE-SHARING SYSTEMS

    OpenAIRE

    Leung, Philip; Svensson, Daniel

    2015-01-01

    The project aims at solving the problem of non-repudiation, integrity and confidentiality of data when digitally exchanging sensitive resources between parties that need to be able to trust each other without the need for a trusted third party. This is done in the framework of answering to what extent digital resources can be shared securely in a decentralized public ledger-based system compared to trust-based alternatives. A background of existing resource sharing solutions is explored which...

  6. Structural Dynamics of Soluble Chloride Intracellular Channel Protein CLIC1 Examined by Amide Hydrogen-Deuterium Exchange Mass Spectrometry (DXMS)†

    Science.gov (United States)

    Stoychev, Stoyan H.; Nathaniel, Christos; Fanucchi, Sylvia; Brock, Melissa; Li, Sheng; Asmus, Kyle; Woods, Virgil L.; Dirr, Heini W.

    2009-01-01

    Chloride intracellular channel protein 1 (CLIC1) functions as an anion channel in plasma and nuclear membranes when its soluble monomeric form converts to an integral-membrane form. The transmembrane region of CLIC1 is located in its thioredoxin-like domain 1 but the mechanism whereby the protein converts to its membrane conformation has yet to be determined. Since channel formation in membranes is enhanced at low pH (5 to 5.5), a condition that is found at the surface of membranes, the structural dynamics of soluble CLIC1 was studied at pH 7 and at pH 5.5 in the absence of membranes by amide hydrogen-deuterium exchange mass spectrometry (DXMS). Rapid hydrogen exchange data indicate that CLIC1 displays a similar core structure at these pH values. Domain 1 is less stable than the all-helical domain 2 and, while the structure of domain 1 remains intact, its conformational flexibility is further increased in an acidic environment (pH 5.5). In the absence of membrane, an acidic environment appears to prime the solution structure of CLIC1 by destabilising domain 1 in order to lower the activation energy barrier for its conversion to the membrane-insertion conformation. The significantly enhanced H/D-exchange rates at pH 5.5 displayed by two segments (peptides 11-31 and 68-82) could be due to the protonation of acidic residues in salt bridges. One of these segments (peptide 11-31) includes part of the transmembrane region which, in the solution structure, consists of helix α1. This helix is intrinsically stable and is most likely retained in the membrane conformation. Strand β2, another element of the transmembrane region, displays a propensity to form a helical structure and has putative N- and C-capping motifs, suggesting that it too most likely forms a helix in a lipid bilayer. PMID:19650640

  7. The decentralization of social assistance and the rise of disability insurance enrolment

    NARCIS (Netherlands)

    Roelofs, Gijs; van Vuuren, Daniël

    Fiscal decentralization, the decentralization of government expenditures to local governments, may enhance public sector efficiency. Vertical externalities, i.e. spillovers between local and central government, could however undo part of this advantage. In this paper we estimate spillovers from

  8. Fiscal Decentralization and Regional Financial Efficiency: An Empirical Analysis of Spatial Durbin Model

    Directory of Open Access Journals (Sweden)

    Jianmin Liu

    2016-01-01

    Full Text Available Based on panel data covering the period from 2003 to 2012 in China’s 281 prefecture-level cities, we use superefficiency SBM model to measure regional financial efficiency and empirically test the spatial effects of fiscal decentralization on regional financial efficiency with SDM. The estimated results indicate that there exist significant spatial spillover effects among regional financial efficiency with the features of time inertia and spatial dependence. The positive promoting effect of fiscal decentralization on financial efficiency in local region depends on the symmetry between fiscal expenditure decentralization and revenue decentralization. Additionally, there exists inconsistency in the spatial effects of fiscal expenditure decentralization and revenue decentralization on financial efficiency in neighboring regions. The negative effect of fiscal revenue decentralization on financial efficiency in neighboring regions is more significant than that of fiscal expenditure decentralization.

  9. Decentralized control of discrete-time linear time invariant systems with input saturation

    NARCIS (Netherlands)

    Deliu, Ciprian; Deliu, C.; Malek, Babak; Roy, Sandip; Saberi, Ali; Stoorvogel, Antonie Arij

    2009-01-01

    We study decentralized stabilization of discrete time linear time invariant (LTI) systems subject to actuator saturation, using LTI controllers. The requirement of stabilization under both saturation constraints and decentralization impose obvious necessary conditions on the open-loop plant, namely

  10. Localization of acoustic sources using a decentralized particle filter

    Directory of Open Access Journals (Sweden)

    Gerstoft Peter

    2011-01-01

    Full Text Available Abstract This paper addresses the decentralized localization of an acoustic source in a (wireless sensor network based on the underlying partial differential equation (PDE. The PDE is transformed into a distributed state-space model and augmented by a source model. Inferring the source state amounts to a non-linear non-Gaussian Bayesian estimation problem for whose solution we implement a decentralized particle filter (PF operating within and across clusters of sensor nodes. The aggregation of the local posterior distributions from all clusters is achieved via an enhanced version of the maximum consensus algorithm. Numerical simulations illustrate the performance of our scheme.

  11. Decentralized Data Sharing of Tissue Microarrays for Investigative Research in Oncology

    Directory of Open Access Journals (Sweden)

    Wenjin Chen

    2006-01-01

    Full Text Available Tissue microarray technology (TMA is a relatively new approach for efficiently and economically assessing protein and gene expression across large ensembles of tissue specimens. Tissue microarray technology holds great potential for reducing the time and cost associated with conducting research in tissue banking, proteomics, and outcome studies. However, the sheer volume of images and other data generated from even limited studies involving tissue microarrays quickly approach the processing capacity and resources of a division or department. This challenge is compounded by the fact that large-scale projects in several areas of modern research rely upon multi-institutional efforts in which investigators and resources are spread out over multiple campuses, cities, and states. To address some of the data management issues several leading institutions have begun to develop their own “in-house” systems, independently, but such data will be only minimally useful if it isn’t accessible to others in the scientifi c community. Investigators at different institutions studying the same or related disorders might benefit from the synergy of sharing results. To facilitate sharing of TMA data across different database implementations, the Technical Standards Committee of the Association for Pathology Informatics organized workshops in efforts to establish a standardized TMA data exchange specification. The focus of our research does not relate to the establishment of standards for exchange, but rather builds on these efforts and concentrates on the design, development and deployment of a decentralized collaboratory for the unsupervised characterization, and seamless and secure discovery and sharing of TMA data. Specifically, we present a self-organizing, peer-to-peer indexing and discovery infrastructure for quantitatively assessing digitized TMA’s. The system utilizes a novel, optimized decentralized search engine that supports flexible querying

  12. Effects of Drying Process on an IgG1 Monoclonal Antibody Using Solid-State Hydrogen Deuterium Exchange with Mass Spectrometric Analysis (ssHDX-MS).

    Science.gov (United States)

    Moussa, Ehab M; Wilson, Nathan E; Zhou, Qi Tony; Singh, Satish K; Nema, Sandeep; Topp, Elizabeth M

    2018-01-03

    Lyophilization and spray drying are widely used to manufacture solid forms of therapeutic proteins. Lyophilization is used to stabilize proteins vulnerable to degradation in solution, whereas spray drying is mainly used to prepare inhalation powders or as an alternative to freezing for storing bulk drug substance. Both processes impose stresses that may adversely affect protein structure, stability and bioactivity. Here, we compared lyophilization with and without controlled ice nucleation, and spray drying for their effects on the solid-state conformation and matrix interactions of a model IgG1 monoclonal antibody (mAb). Solid-state conformation and matrix interactions of the mAb were probed using solid-state hydrogen-deuterium exchange with mass spectrometric analysis (ssHDX-MS), and solid-state Fourier transform infrared (ssFTIR) and solid-state fluorescence spectroscopies. mAb conformation and/or matrix interactions were most perturbed in mannitol-containing samples and the distribution of states was more heterogeneous in sucrose and trehalose samples that were spray dried. The findings demonstrate the sensitivity of ssHDX-MS to changes weakly indicated by spectroscopic methods, and support the broader use of ssHDX-MS to probe formulation and process effects on proteins in solid samples.

  13. Design and Validation of In-Source Atmospheric Pressure Photoionization Hydrogen/Deuterium Exchange Mass Spectrometry with Continuous Feeding of D2O

    Science.gov (United States)

    Acter, Thamina; Lee, Seulgidaun; Cho, Eunji; Jung, Maeng-Joon; Kim, Sunghwan

    2018-01-01

    In this study, continuous in-source hydrogen/deuterium exchange (HDX) atmospheric pressure photoionization (APPI) mass spectrometry (MS) with continuous feeding of D2O was developed and validated. D2O was continuously fed using a capillary line placed on the center of a metal plate positioned between the UV lamp and nebulizer. The proposed system overcomes the limitations of previously reported APPI HDX-MS approaches where deuterated solvents were premixed with sample solutions before ionization. This is particularly important for APPI because solvent composition can greatly influence ionization efficiency as well as the solubility of analytes. The experimental parameters for APPI HDX-MS with continuous feeding of D2O were optimized, and the optimized conditions were applied for the analysis of nitrogen-, oxygen-, and sulfur-containing compounds. The developed method was also applied for the analysis of the polar fraction of a petroleum sample. Thus, the data presented in this study clearly show that the proposed HDX approach can serve as an effective analytical tool for the structural analysis of complex mixtures. [Figure not available: see fulltext.

  14. Different conformational dynamics of PDZ1 and PDZ2 in full-length EBP50 analyzed by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Park, Ji Young; Duc, Nguyen Minh; Kim, Dong Kyun; Lee, Su Youn; Li, Sheng; Seo, Min-Duk; Woods, Virgil L; Chung, Ka Young

    2015-08-01

    Ezrin-radixin-moesin-binding protein 50 (EBP50) is a scaffolding protein expressed in polarized epithelial cells in various organs, including the liver, kidney, and small intestine, in which it regulates the trafficking and targeting cellular proteins. EBP50 contains two postsynaptic density-95/disk-large/ZO-1 homology (PDZ) domains (e.g., PDZ1 and PDZ2) and an ezrin/radixin/moesin-binding (EB) domain. PDZ domains are one of the major scaffolding domains regulating protein-protein interactions with critical biological roles in cell polarity, migration, proliferation, recognition, and cell-cell interaction. PDZ1 and PDZ2 in EBP50 have different ligand selectivity, although several high-resolution structural studies of isolated PDZ1 and PDZ2 showed similar structures. We studied the conformations of full-length EBP50 and isolated PDZ1 and PDZ2 using hydrogen/deuterium exchange mass spectrometry (HDX-MS). The deuterium uptake profiles of isolated PDZ1 and PDZ2 were similar to those of full-length EBP50. Interestingly, PDZ1 was more dynamic than PDZ2, and these PDZ domains underwent different conformational changes upon ligand binding. These results might explain the differences in ligand-selectivity between PDZ1 and PDZ2.

  15. Hydrogen/deuterium exchange mass spectrometry and site-directed disulfide cross-linking suggest an important dynamic interface between the two lysostaphin domains.

    Science.gov (United States)

    Lu, Hai-Rong; Gu, Mei-Gang; Huang, Qiang; Huang, Jin-jiang; Lu, Wan-Ying; Lu, Hong; Huang, Qing-Shan

    2013-04-01

    Lysostaphin is a peptidoglycan hydrolase secreted by Staphylococcus simulans. It can specifically lyse Staphylococcus aureus and is being tested as a novel antibacterial agent. The protein contains an N-terminal catalytic domain and a C-terminal cell wall targeting domain. Although the two domains from homologous enzymes were structurally determined, the structural organization of lysostaphin domains remains unknown. We used hydrogen/deuterium exchange mass spectrometry (H/DX-MS) and site-directed disulfide cross-linking to probe the interface between the lysostaphin catalytic and targeting domains. H/DX-MS-mediated comparison of peptides from full-length lysostaphin and the separated domains identified four peptides of lower solvent accessibility in the full-length protein. Cross-linking analysis using cysteine pair substitutions within those peptides showed that two pairs of cysteines can form disulfide bonds, supporting the domain association role of the targeted peptides. The cross-linked mutant exhibited a binding capacity to S. aureus that was similar to that of the wild-type protein but reduced bacteriolytic activity probably because of restraint in conformation. The diminished activity was further reduced with increasing NaCl concentrations that can cause contractions of bacterial peptidoglycan. The lytic activity, however, could be fully recovered by reducing the disulfide bonds. These results suggest that lysostaphin may require dynamic association of the two domains for coordinating substrate binding and target cleavage on the elastic peptidoglycan. Our study will help develop site-specific PEGylated lysostaphin to treat systemic S. aureus infections.

  16. Sites involved in intra- and interdomain allostery associated with the activation of factor VIIa pinpointed by hydrogen-deuterium exchange and electron transfer dissociation mass spectrometry.

    Science.gov (United States)

    Song, Hongjian; Olsen, Ole H; Persson, Egon; Rand, Kasper D

    2014-12-19

    Factor VIIa (FVIIa) is a trypsin-like protease that plays an important role in initiating blood coagulation. Very limited structural information is available for the free, inactive form of FVIIa that circulates in the blood prior to vascular injury and the molecular details of its activity enhancement remain elusive. Here we have applied hydrogen/deuterium exchange mass spectrometry coupled to electron transfer dissociation to pinpoint individual residues in the heavy chain of FVIIa whose conformation and/or local interaction pattern changes when the enzyme transitions to the active form, as induced either by its cofactor tissue factor or a covalent active site inhibitor. Identified regulatory residues are situated at key sites across one continuous surface of the protease domain spanning the TF-binding helix across the activation pocket to the calcium binding site and are embedded in elements of secondary structure and at the base of flexible loops. Thus these residues are optimally positioned to mediate crosstalk between functional sites in FVIIa, particularly the cofactor binding site and the active site. Our results unambiguously show that the conformational allosteric activation signal extends to the EGF1 domain in the light chain of FVIIa, underscoring a remarkable intra- and interdomain allosteric regulation of this trypsin-like protease. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  17. Conformational dynamics of the bovine mitochondrial ADP/ATP carrier isoform 1 revealed by hydrogen/deuterium exchange coupled to mass spectrometry.

    Science.gov (United States)

    Rey, Martial; Man, Petr; Clémençon, Benjamin; Trézéguet, Véronique; Brandolin, Gérard; Forest, Eric; Pelosi, Ludovic

    2010-11-05

    The mitochondrial adenine nucleotide carrier (Ancp) catalyzes the transport of ADP and ATP across the mitochondrial inner membrane, thus playing an essential role in cellular energy metabolism. During the transport mechanism the carrier switches between two different conformations that can be blocked by two toxins: carboxyatractyloside (CATR) and bongkrekic acid. Therefore, our understanding of the nucleotide transport mechanism can be improved by analyzing structural differences of the individual inhibited states. We have solved the three-dimensional structure of bovine carrier isoform 1 (bAnc1p) in a complex with CATR, but the structure of the carrier-bongkrekic acid complex, and thus, the detailed mechanism of transport remains unknown. Improvements in sample processing in the hydrogen/deuterium exchange technique coupled to mass spectrometry (HDX-MS) have allowed us to gain novel insights into the conformational changes undergone by bAnc1p. This paper describes the first study of bAnc1p using HDX-MS. Results obtained with the CATR-bAnc1p complex were fully in agreement with published results, thus, validating our approach. On the other hand, the HDX kinetics of the two complexes displays marked differences. The bongkrekic acid-bAnc1p complex exhibits greater accessibility to the solvent on the matrix side, whereas the CATR-bAnc1p complex is more accessible on the intermembrane side. These results are discussed with respect to the structural and biochemical data available on Ancp.

  18. Mechanism and kinetics of tyrosinase inhibition by glycolic acid: a study using conventional spectroscopy methods and hydrogen/deuterium exchange coupling with mass spectrometry.

    Science.gov (United States)

    Ma, Da; Tu, Zong-Cai; Wang, Hui; Zhang, Lu; He, Na; McClements, David Julian

    2017-01-25

    Tyrosinase is an enzyme that promotes enzymatic browning of fruits and vegetables, thereby reducing product quality. A variety of analytical tools were used to characterize the interactions between tyrosinase and a natural tyrosinase inhibitor (glycolic acid). Hydrogen/deuterium exchange coupling with mass spectrometry (HDX-MS) was used to elucidate the interaction mechanism between glycolic acid and tyrosinase. UV-visible, fluorescence and circular dichroism spectroscopy analysis indicated that glycolic acid inhibited tyrosinase activity in a mixed-type manner with an IC 50 of 83 ± 14 μM. The results of these techniques suggested that glycolic acid bound to tyrosinase through hydrophobic attraction, and this interaction led to a pronounced conformational change of the enzyme molecules. HDX-MS analysis showed that the activity of tyrosinase was primarily inhibited by a structural perturbation of its active site (His 263). This study provides a comprehensive understanding of the interaction between glycolic acid and tyrosinase, which could lead to new approaches to control tyrosinase activity in foods and other products.

  19. Identification of pharmacological chaperones for Gaucher disease and characterization of their effects on beta-glucocerebrosidase by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Tropak, Michael B; Kornhaber, Gregory J; Rigat, Brigitte A; Maegawa, Gustavo H; Buttner, Justin D; Blanchard, Jan E; Murphy, Cecilia; Tuske, Steven J; Coales, Stephen J; Hamuro, Yoshitomo; Brown, Eric D; Mahuran, Don J

    2008-11-03

    Point mutations in beta-glucocerebrosidase (GCase) can result in a deficiency of both GCase activity and protein in lysosomes thereby causing Gaucher Disease (GD). Enzyme inhibitors such as isofagomine, acting as pharmacological chaperones (PCs), increase these levels by binding and stabilizing the native form of the enzyme in the endoplasmic reticulum (ER), and allow increased lysosomal transport of the enzyme. A high-throughput screen of the 50,000-compound Maybridge library identified two, non-carbohydrate-based inhibitory molecules, a 2,4-diamino-5-substituted quinazoline (IC(50) 5 microM) and a 5-substituted pyridinyl-2-furamide (IC(50) 8 microM). They raised the levels of functional GCase 1.5-2.5-fold in N370S or F213I GD fibroblasts. Immunofluorescence confirmed that treated GD fibroblasts had decreased levels of GCase in their ER and increased levels in lysosomes. Changes in protein dynamics, monitored by hydrogen/deuterium-exchange mass spectrometry, identified a domain III active-site loop (residues 243-249) as being significantly stabilized upon binding of isofagomine or either of these two new compounds; this suggests a common mechanism for PC enhancement of intracellular transport.

  20. Using hydrogen deuterium exchange mass spectrometry to engineer optimized constructs for crystallization of protein complexes: Case study of PI4KIIIβ with Rab11.

    Science.gov (United States)

    Fowler, Melissa L; McPhail, Jacob A; Jenkins, Meredith L; Masson, Glenn R; Rutaganira, Florentine U; Shokat, Kevan M; Williams, Roger L; Burke, John E

    2016-04-01

    The ability of proteins to bind and interact with protein partners plays fundamental roles in many cellular contexts. X-ray crystallography has been a powerful approach to understand protein-protein interactions; however, a challenge in the crystallization of proteins and their complexes is the presence of intrinsically disordered regions. In this article, we describe an application of hydrogen deuterium exchange mass spectrometry (HDX-MS) to identify dynamic regions within type III phosphatidylinositol 4 kinase beta (PI4KIIIβ) in complex with the GTPase Rab11. This information was then used to design deletions that allowed for the production of diffraction quality crystals. Importantly, we also used HDX-MS to verify that the new construct was properly folded, consistent with it being catalytically and functionally active. Structures of PI4KIIIβ in an Apo state and bound to the potent inhibitor BQR695 in complex with both GTPγS and GDP loaded Rab11 were determined. This hybrid HDX-MS/crystallographic strategy revealed novel aspects of the PI4KIIIβ-Rab11 complex, as well as the molecular mechanism of potency of a PI4K specific inhibitor (BQR695). This approach is widely applicable to protein-protein complexes, and is an excellent strategy to optimize constructs for high-resolution structural approaches. © 2016 The Protein Society.

  1. An Asymmetric Runaway Domain Swap Antithrombin Dimer as a Key Intermediate for Polymerization Revealed by Hydrogen/Deuterium-Exchange Mass Spectrometry.

    Science.gov (United States)

    Trelle, Morten Beck; Pedersen, Shona; Østerlund, Eva Christina; Madsen, Jeppe Buur; Kristensen, Søren Risom; Jørgensen, Thomas J D

    2017-01-03

    Antithrombin deficiency is associated with increased risk of venous thrombosis. In certain families, this condition is caused by pathogenic polymerization of mutated antithrombin in the blood. To facilitate future development of pharmaceuticals against antithrombin polymerization, an improved understanding of the polymerogenic intermediates is crucial. However, X-ray crystallography of these intermediates is severely hampered by the difficulty in obtaining well-diffracting crystals of transient and heterogeneous noncovalent protein assemblies. Furthermore, their large size prohibits structural analysis by NMR spectroscopy. Here, we show how hydrogen/deuterium-exchange mass spectrometry (HDX-MS) provides detailed insight into the structural dynamics of each subunit in a polymerization-competent antithrombin dimer. Upon deuteration, this dimer surprisingly yields bimodal isotope distributions for the majority of peptides, demonstrating an asymmetric configuration of the two subunits. The data reveal that one subunit is very dynamic, potentially intrinsically disordered, whereas the other is considerably less dynamic. The local subunit-specific deuterium uptake of this polymerization-competent dimer strongly supports a β4A-β5A β-hairpin runaway domain swap mechanism for antithrombin polymerization. HDX-MS thus holds exceptional promise as an enabling analytical technique in the efforts toward future pharmacological intervention with protein polymerization and associated diseases.

  2. Hydrogen/deuterium exchange mass spectrometry and computational modeling reveal a discontinuous epitope of an antibody/TL1A Interaction.

    Science.gov (United States)

    Huang, Richard Y-C; Krystek, Stanley R; Felix, Nathan; Graziano, Robert F; Srinivasan, Mohan; Pashine, Achal; Chen, Guodong

    2018-01-01

    TL1A, a tumor necrosis factor-like cytokine, is a ligand for the death domain receptor DR3. TL1A, upon binding to DR3, can stimulate lymphocytes and trigger secretion of proinflammatory cytokines. Therefore, blockade of TL1A/DR3 interaction may be a potential therapeutic strategy for autoimmune and inflammatory diseases. Recently, the anti-TL1A monoclonal antibody 1 (mAb1) with a strong potency in blocking the TL1A/DR3 interaction was identified. Here, we report on the use of hydrogen/deuterium exchange mass spectrometry (HDX-MS) to obtain molecular-level details of mAb1's binding epitope on TL1A. HDX coupled with electron-transfer dissociation MS provided residue-level epitope information. The HDX dataset, in combination with solvent accessible surface area (SASA) analysis and computational modeling, revealed a discontinuous epitope within the predicted interaction interface of TL1A and DR3. The epitope regions span a distance within the approximate size of the variable domains of mAb1's heavy and light chains, indicating it uses a unique mechanism of action to block the TL1A/DR3 interaction.

  3. Fast analysis of quaternary ammonium pesticides in food and beverages using cation-exchange chromatography coupled with isotope-dilution high-resolution mass spectrometry.

    Science.gov (United States)

    Nardin, Tiziana; Barnaba, Chiara; Abballe, Franco; Trenti, Gianmaria; Malacarne, Mario; Larcher, Roberto

    2017-10-01

    A fast separation based on cation-exchange liquid chromatography coupled with high-resolution mass spectrometry is proposed for simultaneous determination of chlormequat, difenzoquat, diquat, mepiquat and paraquat in several food and beverage commodities. Solid samples were extracted using a mixture of water/methanol/formic acid (69.6:30:0.4, v/v/v), while liquid samples were ten times diluted with the same solution. Separation was carried out on an experimental length-modified IonPac CS17 column (2 × 15 mm 2 ) that allowed the use of formic acid and acetonitrile as mobile phase. Detection limits for food and beverage matrices were established at 1.5 μg/L for chlormequat, difenzoquat and mepiquat, and 3 μg/L for diquat and paraquat, while for drinking water a pre-analytical sample concentration allowed detection limits of 9 and 20 ng/L, respectively. Precision, as repeatability (RSD%), ranged from 0.2 to 24%, with a median value of 6%, and trueness, as recovery, ranged from 64 to 118%, with a median value of 96%. The method developed was successfully applied to investigate the presence of herbicide residues in commercial commodities (mineral water, orange juice, beer, tea, green coffee bean, toasted coffee powder, cocoa bean, white corn flour, rice and sugar samples). © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Hydrogen/deuterium exchange and mass spectrometric analysis of a protein containing multiple disulfide bonds: Solution structure of recombinant macrophage colony stimulating factor-beta (rhM-CSFβ)

    Science.gov (United States)

    Yan, Xuguang; Zhang, Heidi; Watson, Jeffrey; Schimerlik, Michael I.; Deinzer, Max L.

    2002-01-01

    Studies with the homodimeric recombinant human macrophage colony-stimulating factor beta (rhM-CSFβ), show for the first time that a large number (9) of disulfide linkages can be reduced after amide hydrogen/deuterium (H/D) exchange, and the protein digested and analyzed successfully for the isotopic composition by electrospray mass spectrometry. Analysis of amide H/D after exchange-in shows that in solution the conserved four-helix bundle of (rhM-CSFβ) has fast and moderately fast exchangeable sections of amide hydrogens in the αA helix, and mostly slow exchanging sections of amide hydrogens in the αB, αC, and αD helices. Most of the amide hydrogens in the loop between the β1 and β4 sheets exhibited fast or moderately fast exchange, whereas in the amino acid 63–67 loop, located at the interface of the two subunits, the exchange was slow. Solvent accessibility as measured by H/D exchange showed a better correlation with the average depth of amide residues calculated from reported X-ray crystallographic data for rhM-CSFα than with the average B-factor. The rates of H/D exchange in rhM-CSFβ appear to correlate well with the exposed surface calculated for each amino acid residue in the crystal structure except for the αD helix. Fast hydrogen isotope exchange throughout the segment amino acids 150–221 present in rhM-CSFβ, but not rhM-CSFα, provides evidence that the carboxy-terminal region is unstructured. It is, therefore, proposed that the anomalous behavior of the αD helix is due to interaction of the carboxy-terminal tail with this helical segment. PMID:12192067

  5. Decentralization and basic services provision: water and sanitation in Ghana

    NARCIS (Netherlands)

    M.P. van Dijk (Meine Pieter)

    2007-01-01

    textabstractHow does decentralization reform affect the provision of basic services, particularly water and sanitation in Ghana? We deal in particular the delivery of water and sanitation, given the importance of water and sanitation. Conclusions are drawn concerning the policy implications for the

  6. Canarsie and Fuentes: The Limits of School Decentralization

    Science.gov (United States)

    Ravitch, Diane

    1973-01-01

    The Fuentes case demonstrated how the community's rights could be subverted by a purposeful faction; the Canarsie case showed how unchecked local control could damage children's rights. Successful decentralization requires the central board to assert its authority to prevent the arbitrary exercise of local power. (EH)

  7. Contesting sharia : state law, decentralization and Minangkabau custom

    NARCIS (Netherlands)

    Huda, Yasrul

    2013-01-01

    This book explains how Sharia, commonly called Perda Sharia (Sharia by-law) in Indonesia, was legislated on the provincial, regional and municipal level in West Sumatra. This process began after the government started a decentralization policy in 2000. Although the law of local autonomy prescribes

  8. A simulation model of a coordinated decentralized linear supply chain

    NARCIS (Netherlands)

    Ashayeri, Jalal; Cannella, S.; Lopez Campos, M.; Miranda, P.A.

    2015-01-01

    This paper presents a simulation-based study of a coordinated, decentralized linear supply chain (SC) system. In the proposed model, any supply tier considers its successors as part of its inventory system and generates replenishment orders on the basis of its partners’ operational information. We

  9. Decentralization, Local Rights and the Construction of Women's ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Kenya, Tanzania and Uganda have adopted new land laws, policies and institutional arrangements to accommodate decentralization of land administration and management. Land is a significant economic resource in a predominantly agrarian society, one in which women as farmers make an immense contribution.

  10. Video - Real Rights: Decentralization and Women in South Asia ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    2010-10-20

    Oct 20, 2010 ... But has shifting more power and resources to the local level really improved women's lives? These videos show the results of fieldwork supported by IDRC's Women's Rights and Citizenship program, which aimed to learn from women's experiences of decentralization in Nepal, Pakistan, and India.

  11. Independence and Collaboration; Why We Should Decentralize Writing Centers.

    Science.gov (United States)

    Smith, Louise Z.

    1986-01-01

    Notes the inevitable tensions that arise between centripetal writing centers and centrifugal writing across the curriculum programs. Examines the tutoring program at an eastern university as an example of a decentralized writing center that resists pressures to assume a uniform composition pedagogy and coordinates its work with many parts of the…

  12. Reaction Diffusion and Chemotaxis for Decentralized Gathering on FPGAs

    Directory of Open Access Journals (Sweden)

    Bernard Girau

    2009-01-01

    and rapid simulations of the complex dynamics of this reaction-diffusion model. Then we describe the FPGA implementation of the environment together with the agents, to study the major challenges that must be solved when designing a fast embedded implementation of the decentralized gathering model. We analyze the results according to the different goals of these hardware implementations.

  13. Decentralized Urban Solid Waste Management in Indonesia | CRDI ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    This project will develop and validate four models of decentralized solid waste management in low-income urban areas of Tangerang, Sidoarjo, Denpasar and Mataram. The four models will be evaluated in terms of cost-recovery, technical sustainability and social acceptability. Efforts will be made to achieve recognition of ...

  14. Centralized vs. De-centralized Multinationals and Taxes

    DEFF Research Database (Denmark)

    Nielsen, Søren Bo; Raimondos-Møller, Pascalis; Schjelderup, Guttorm

    2005-01-01

    The paper examines how country tax differences affect a multinational enterprise's choice to centralize or de-centralize its decision structure. Within a simple model that emphasizes the multiple conflicting roles of transfer prices in MNEs - here, as a strategic pre-commitment device and a tax...

  15. DECENTRALIZATION OF MUNICIPAL SERVICES – LEARNING BY DOING

    Directory of Open Access Journals (Sweden)

    Cristina Elena NICOLESCU

    2017-05-01

    Full Text Available Public services decentralization is a major concern for policy makers when it comes to identifying the optimum model for reorganizing these services, in light of the 3Es of the organizational performance. The field experiences show that this process is different both from one state to another, and depending on the targeted activity sector, out of which the local transport service is distinguished as an ‘institutional orphan’. Taking into account one of the smart-cities’ recognition criteria, the urban mobility, the paper aims at substantiating that, despite the specific incrementalism of the public services decentralization, having a negative impact upon the services’ efficiency, in the case of local transport service, recognizing the right to mobility and the need to ensuring the environment for exercising this right, impels the ‘bureaucratic apparatus’ to accelerate and consolidate the decentralization of this service. Therefore, the paper puts forward a case study on the impact of decentralization upon the local public transport service of Bucharest municipality.

  16. Spectrum Allocation for Decentralized Transmission Strategies: Properties of Nash Equilibria

    Directory of Open Access Journals (Sweden)

    Peter von Wrycza

    2009-01-01

    Full Text Available The interaction of two transmit-receive pairs coexisting in the same area and communicating using the same portion of the spectrum is analyzed from a game theoretic perspective. Each pair utilizes a decentralized iterative water-filling scheme to greedily maximize the individual rate. We study the dynamics of such a game and find properties of the resulting Nash equilibria. The region of achievable operating points is characterized for both low- and high-interference systems, and the dependence on the various system parameters is explicitly shown. We derive the region of possible signal space partitioning for the iterative water-filling scheme and show how the individual utility functions can be modified to alter its range. Utilizing global system knowledge, we design a modified game encouraging better operating points in terms of sum rate compared to those obtained using the iterative water-filling algorithm and show how such a game can be imitated in a decentralized noncooperative setting. Although we restrict the analysis to a two player game, analogous concepts can be used to design decentralized algorithms for scenarios with more players. The performance of the modified decentralized game is evaluated and compared to the iterative water-filling algorithm by numerical simulations.

  17. Centralization and Decentralization of Schools' Physical Facilities Management in Nigeria

    Science.gov (United States)

    Ikoya, Peter O.

    2008-01-01

    Purpose: This research aims to examine the difference in the availability, adequacy and functionality of physical facilities in centralized and decentralized schools districts, with a view to making appropriate recommendations to stakeholders on the reform programmes in the Nigerian education sector. Design/methodology/approach: Principals,…

  18. Conservation and Livelihood Impacts of Decentralized Forest Governance in Ethiopia

    DEFF Research Database (Denmark)

    Yietagesu, Aklilu Ameha

    The dissertation focuses on a specific form of forest decentralization, participatory forest management (PFM). The underlying premise of PFM is that sustainable forest management is most likely to occur when local communities manage local forests, and when they get access to direct benefits from...

  19. Decentralized Urban Solid Waste Management in Indonesia | IDRC ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    This project will develop and validate four models of decentralized solid waste management in low-income urban areas of Tangerang, Sidoarjo, Denpasar and Mataram. The four models will be evaluated in terms of cost-recovery, technical sustainability and social acceptability. Efforts will be made to achieve recognition of ...

  20. Decentralization, Local Politics and the Construction of Women's ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    This project constitutes the first phase of a comparative study of decentralization, local politics and the construction of women's citizenship in Uganda, Kenya and Tanzania. ... The Science Granting Councils Initiative in sub-Saharan Africa received a Science Diplomacy Award from the Government of South Africa.

  1. Decentralized Receding Horizon Control and Coordination of Autonomous Vehicle Formations

    NARCIS (Netherlands)

    Keviczky, T.; Borelli, F.; Fregene, K.; Godbole, D.; Bals, G.J.

    2008-01-01

    This paper describes the application of a novel methodology for high-level control and coordination of autonomous vehicle teams and its demonstration on high-fidelity models of the organic air vehicle developed at Honeywell Laboratories. The scheme employs decentralized receding horizon controllers

  2. Dynamic orders of decentralized H_inf controllers

    DEFF Research Database (Denmark)

    Stoustrup, Jakob; Niemann, Hans Henrik

    1999-01-01

    The problem of decentralized control is addressed, i.e. the problem of designing a controller where each control input is allowed to use only some of the measurements. It is shown that, for such problems, there does not always exist a sequence of controllers of bounded order which obtains near-op...

  3. Dynamical orders of decentralized H-infinity controllers

    DEFF Research Database (Denmark)

    Stoustrup, Jakob; Niemann, H.H.

    1999-01-01

    The problem of decentralized control is addressed, i.e. the problem of designing a controller where each control input is allowed to use only some of the measurements. It is shown that, for such problems, there does not always exist a sequence of controllers of bounded order which obtains near-op...

  4. A Logic for Auditing Accountability in Decentralized Systems

    NARCIS (Netherlands)

    Corin, R.J.; Etalle, Sandro; den Hartog, Jeremy; Lenzini, Gabriele; Staicu, I.

    We propose a language that allows agents to distribute data with usage policies in a decentralized architecture. In our framework, the compliance with usage policies is not enforced. However, agents may be audited by an authority at an arbitrary moment in time. We design a logic that allows audited

  5. A Decentralized Storage Strategy for Residential Feeders with Photovoltaics

    DEFF Research Database (Denmark)

    Marra, Francesco; Yang, Guangya; Træholt, Chresten

    2014-01-01

    This paper proposes a decentralized voltage support strategy for LV residential feeders with high roof-top PV capacity installed. The proposed strategy is capable of increasing the local consumption at private households with PV during high generation periods, by the use of locally controlled dom...

  6. The influence of decentralization on effectiveness of extension ...

    African Journals Online (AJOL)

    Against the background of frequent organisational changes and restructuring, often based on impulsive decisions rather than structured feasibility studies or evaluations, this article examines the influence of decentralization on the performance of an extension organization. Based on a survey of 353 respondents from ...

  7. Formula Funding of Schools, Decentralization and Corruption: A Comparative Analysis

    Science.gov (United States)

    Levacic, Rosalind; Downes, Peter

    2004-01-01

    This study sets out to examine the relationship, if any, between the decentralization of funding for schools and the prevalence of corruption and fraud. It is based upon work carried out in four countries: Australia (the State of Victoria), the United Kingdom (specifically England), Poland (with particular reference to two cities) and Brazil…

  8. Decentralizing Education in Guatemala : School Management by Local Communities

    OpenAIRE

    Carlos Rojas; Alexandria Valerio; Angela Demas

    2005-01-01

    Guatemala set out in 1992 to increase access to education in remote areas. Its National Community-managed Program for Educational Development (PRONADE) has evolved from a small, innovative pilot program in 19 rural communities, to a nationwide program reaching over 4,100 communities and 445,000 children. PRONADE is one of the most proactive managerial, administrative, and financial decentr...

  9. The Relationship Between Traffic Stability and Capacity for Decentralized Airspace

    NARCIS (Netherlands)

    Sunil, E.; Maas, J.B.; Ellerbroek, J.; Hoekstra, J.M.; Tra, M.A.P.

    2016-01-01

    The work that is presented in this paper is part of an ongoing study on the relationship between structure and capacity of decentralized airspace concepts. In this paper, the effect of traffic stability, which considers the occurrence of conflict chain reactions as a result of conflict resolution

  10. Decentralized supervisory control of discrete-event systems with communication

    NARCIS (Netherlands)

    K.C. Wong; J.H. van Schuppen (Jan)

    1996-01-01

    textabstractThis paper formulates a problem of communication channels in decentralized control. A necessary and sufficient condition is obtained for the existence of such a channel and is illustrated with an example of communication protocols. The existence of minimal communication channels is also

  11. Electrical Load Survey and Forecast for a Decentralized Hybrid ...

    African Journals Online (AJOL)

    This paper presents electrical load survey and forecast for a typical off-grid rural decentralized hybrid power generating systems of a rural area (Elebu). With the data available, energy demand and forecast for ten years was estimated. It was also observed that this community has potential for small hydro, wind and solar ...

  12. Analysis of Decentralized Control for Absorption Cycle Heat Pumps

    DEFF Research Database (Denmark)

    Vinther, Kasper; Just Nielsen, Rene; Nielsen, Kirsten Mølgaard

    2015-01-01

    Email Print Request Permissions This paper investigates decentralized control structures for absorption cycle heat pumps and a dynamic nonlinear model of a single-effect LiBr-water absorption system is used as case study. The model has four controllable inputs, which can be used to stabilize...

  13. Decentralization of operating reactor licensing reviews: NRR Pilot Program

    International Nuclear Information System (INIS)

    Hannon, J.N.

    1984-07-01

    This report, which has incorporated comments received from the Commission and ACRS, describes the program for decentralization of selected operating reactor licensing technical review activities. The 2-year pilot program will be reviewed to verify that safety is enhanced as anticipated by the incorporation of prescribed management techniques and application of resources. If the program fails to operate as designed, it will be terminated

  14. Integrating Collaborative and Decentralized Models to Support Ubiquitous Learning

    Science.gov (United States)

    Barbosa, Jorge Luis Victória; Barbosa, Débora Nice Ferrari; Rigo, Sandro José; de Oliveira, Jezer Machado; Rabello, Solon Andrade, Jr.

    2014-01-01

    The application of ubiquitous technologies in the improvement of education strategies is called Ubiquitous Learning. This article proposes the integration between two models dedicated to support ubiquitous learning environments, called Global and CoolEdu. CoolEdu is a generic collaboration model for decentralized environments. Global is an…

  15. Distributed and decentralized control architectures for converter-interfaced microgrids

    DEFF Research Database (Denmark)

    Dragicevic, Tomislav; Wu, Dan; Shafiee, Qobad

    2017-01-01

    This paper gives a summary on recently available technologies for decentralized and distributed control of microgrids. They can be classified into two general categories: 1) power line communication based architectures and 2) multi-agent based architectures. The essential control methods...

  16. Users' perspectives on decentralized rural water services in Tanzania

    NARCIS (Netherlands)

    Masanyiwa, Z.S.; Niehof, A.; Termeer, C.J.A.M.

    2015-01-01

    This article examines the impact of decentralization reforms on improving access to domestic water supply in the rural districts of Kondoa and Kongwa, Tanzania, using a users' and a gender perspective. The article addresses the question whether and to what extent the delivery of gender-sensitive

  17. Attachment for mass exchange apparatus

    Energy Technology Data Exchange (ETDEWEB)

    Maksimenko, M.Z.; Maksimenko, M.M.; Prokopov, O.I.

    1982-01-01

    An attachment is proposed which contains a cylindrical housing in which the windows are profiled with inward bent windows. In order to improve the effectiveness of the attachment by increasing its surface, and mobility, the ends of the housing are equipped with inner annular partitions with radial cross sections. The sections of the partitions between them are bent inwards towards the forming housing in series, from the side of each section. The inward-bent sections of the partitions and the edges are directed to one side or to different sides.

  18. Distributed and decentralized state estimation in gas networks as distributed parameter systems.

    Science.gov (United States)

    Ahmadian Behrooz, Hesam; Boozarjomehry, R Bozorgmehry

    2015-09-01

    In this paper, a framework for distributed and decentralized state estimation in high-pressure and long-distance gas transmission networks (GTNs) is proposed. The non-isothermal model of the plant including mass, momentum and energy balance equations are used to simulate the dynamic behavior. Due to several disadvantages of implementing a centralized Kalman filter for large-scale systems, the continuous/discrete form of extended Kalman filter for distributed and decentralized estimation (DDE) has been extended for these systems. Accordingly, the global model is decomposed into several subsystems, called local models. Some heuristic rules are suggested for system decomposition in gas pipeline networks. In the construction of local models, due to the existence of common states and interconnections among the subsystems, the assimilation and prediction steps of the Kalman filter are modified to take the overlapping and external states into account. However, dynamic Riccati equation for each subsystem is constructed based on the local model, which introduces a maximum error of 5% in the estimated standard deviation of the states in the benchmarks studied in this paper. The performance of the proposed methodology has been shown based on the comparison of its accuracy and computational demands against their counterparts in centralized Kalman filter for two viable benchmarks. In a real life network, it is shown that while the accuracy is not significantly decreased, the real-time factor of the state estimation is increased by a factor of 10. Copyright © 2015 ISA. Published by Elsevier Ltd. All rights reserved.

  19. Exchange transfusion

    Science.gov (United States)

    ... count in a newborn (neonatal polycythemia) Rh-induced hemolytic disease of the newborn Severe disturbances in body chemistry Severe newborn jaundice ... exchange transfusion was performed to treat. Alternative Names Hemolytic disease - exchange transfusion Patient ... Exchange transfusion - series References Costa ...

  20. Decentralizing conservation and diversifying livelihoods within Kanchenjunga Conservation Area, Nepal.

    Science.gov (United States)

    Parker, Pete; Thapa, Brijesh; Jacob, Aerin

    2015-12-01

    To alleviate poverty and enhance conservation in resource dependent communities, managers must identify existing livelihood strategies and the associated factors that impede household access to livelihood assets. Researchers increasingly advocate reallocating management power from exclusionary central institutions to a decentralized system of management based on local and inclusive participation. However, it is yet to be shown if decentralizing conservation leads to diversified livelihoods within a protected area. The purpose of this study was to identify and assess factors affecting household livelihood diversification within Nepal's Kanchenjunga Conservation Area Project, the first protected area in Asia to decentralize conservation. We randomly surveyed 25% of Kanchenjunga households to assess household socioeconomic and demographic characteristics and access to livelihood assets. We used a cluster analysis with the ten most common income generating activities (both on- and off-farm) to group the strategies households use to diversify livelihoods, and a multinomial logistic regression to identify predictors of livelihood diversification. We found four distinct groups of household livelihood strategies with a range of diversification that directly corresponded to household income. The predictors of livelihood diversification were more related to pre-existing socioeconomic and demographic factors (e.g., more landholdings and livestock, fewer dependents, receiving remittances) than activities sponsored by decentralizing conservation (e.g., microcredit, training, education, interaction with project staff). Taken together, our findings indicate that without direct policies to target marginalized groups, decentralized conservation in Kanchenjunga will continue to exclude marginalized groups, limiting a household's ability to diversify their livelihood and perpetuating their dependence on natural resources. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. DECENTRALIZATION AND LOCAL ECONOMIC DEVELOPMENT IN THE KNOWLEDGE SOCIETY

    Directory of Open Access Journals (Sweden)

    Elena, RUSU (CIGU

    2014-11-01

    Full Text Available The sustainable development became a goal for knowledge society, but one of the main question in literature remains as how close is the relationship between economic dvelopment, knowledge society and local governments' level. Given the great diversity of situations and frameworks both across countries and within them in the European Union, I consider that each country must have own policies designed to implement process of decentralization with the commitment of "healthy" economy in a society dominated by the technology and innovation. In this context, the problem occurs in decentralization should be managed on two levels: on the one hand in terms of central-local budgets relations, and on the other hand, in the local public finance structural plan. The paper will try to emphasize the connection between decentralization, local economic sustainable development and knowledge society, taking into account an overview of this three aspects and empirical evidences. In this regard, I will use the background offered by literature and the official statiscal data for analysis to identify the variables which explain decentralization and local economic development in the knowledge society. I estimate the analysis to confirm the hypothesis that there are good aspects, but also deficiencies that require solutions and budgetary policy options as part undisputed positioning local government finance as an engine of development of the whole nation. I consider that the paper can be a useful viewpoint in understanding local public finances in decentralization, which allows researchers to include other sources of information for researching an in a much more complex approach.

  2. Gas-phase ion/molecule isotope-exchange reactions: methodology for counting hydrogen atoms in specific organic structural environments by chemical ionization mass spectrometry

    International Nuclear Information System (INIS)

    Hunt, D.F.; Sethi, S.K.

    1980-01-01

    Ion/molecule reactions are described which facilitate exchange of hydrogens for deuteriums in a variety of different chemical environments. Aromatic hydrogens in alkylbenzenes, oxygenated benzenes, m-toluidine, m-phenylenediamine, thiophene, and several polycyclic aromatic hydrocarbons and metallocenes are exchanged under positive ion CI conditions by using either D 2 O, EtOD, or ND 3 as the reagent gas. Aromatic hydrogens, benzylic hydrogens, and hydrogens on carbon adjacent to carbonyl groups suffer exchange under negative ion CI conditions in ND 3 , D 2 O, and EtOD, respectively. A possible mechanism for the exchange process is discussed. 1 figure, 2 tables

  3. Least-cost network evaluation of centralized and decentralized contributions to global electrification

    International Nuclear Information System (INIS)

    Levin, Todd; Thomas, Valerie M.

    2012-01-01

    The choice between centralized and decentralized electricity generation is examined for 150 countries as a function of population distribution, electricity consumption, transmission cost, and the cost difference between decentralized and centralized electricity generation. A network algorithm is developed to find the shortest centralized transmission network that spans a given fraction of the population in a country. The least-cost combination of centralized and decentralized electricity that serves the country is determined. Case studies of Botswana, Uganda, and Bangladesh illustrate situations that are more and less suited for decentralized electrification. Specific maps for centralized and decentralized generation are presented to show how the least-cost option varies with the relative costs of centralized and decentralized generation and transmission cost. Centralized and decentralized fractions are calculated for 150 countries. For most of the world's population, centralized electricity is the least-cost option. For a number of countries, particularly in Africa, substantial populations and regions may be most cost-effectively served by decentralized electricity. - Highlights: ► Centralized and decentralized electrification are compared for 150 countries. ► A cost-optimized network algorithm finds the least-cost electrification system. ► Least-cost infrastructures combine centralized and decentralized portions. ► For most people, centralized electricity is cheapest option. ► In much of Africa, decentralized electricity may be cheaper than centralized.

  4. Approach to characterization of the higher order structure of disulfide-containing proteins using hydrogen/deuterium exchange and top-down mass spectrometry.

    Science.gov (United States)

    Wang, Guanbo; Kaltashov, Igor A

    2014-08-05

    Top-down hydrogen/deuterium exchange (HDX) with mass spectrometric (MS) detection has recently matured to become a potent biophysical tool capable of providing valuable information on higher order structure and conformational dynamics of proteins at an unprecedented level of structural detail. However, the scope of the proteins amenable to the analysis by top-down HDX MS still remains limited, with the protein size and the presence of disulfide bonds being the two most important limiting factors. While the limitations imposed by the physical size of the proteins gradually become more relaxed as the sensitivity, resolution and dynamic range of modern MS instrumentation continue to improve at an ever accelerating pace, the presence of the disulfide linkages remains a much less forgiving limitation even for the proteins of relatively modest size. To circumvent this problem, we introduce an online chemical reduction step following completion and quenching of the HDX reactions and prior to the top-down MS measurements of deuterium occupancy of individual backbone amides. Application of the new methodology to the top-down HDX MS characterization of a small (99 residue long) disulfide-containing protein β2-microglobulin allowed the backbone amide protection to be probed with nearly a single-residue resolution across the entire sequence. The high-resolution backbone protection pattern deduced from the top-down HDX MS measurements carried out under native conditions is in excellent agreement with the crystal structure of the protein and high-resolution NMR data, suggesting that introduction of the chemical reduction step to the top-down routine does not trigger hydrogen scrambling either during the electrospray ionization process or in the gas phase prior to the protein ion dissociation.

  5. Epitope mapping of inhibitory antibodies targeting the C2 domain of coagulation factor VIII by hydrogen-deuterium exchange mass spectrometry

    Science.gov (United States)

    Sevy, Alexander M.; Healey, John F.; Deng, Wei; Spiegel, P. Clint; Meeks, Shannon L.; Li, Renhao

    2014-01-01

    Summary Background The development of anti-factor VIII (fVIII) antibodies (inhibitors) is a significant complication in the management of patients with hemophilia A, leading to significant increases in morbidity and treatment cost. Using a panel of anti-fVIII monoclonal antibodies (MAbs) to different epitopes on fVIII, we recently have shown that epitope specificity, inhibitor kinetics, and time to maximum inhibition are more important than inhibitor titer in predicting response to fVIII and the combination of fVIII and recombinant factor VIIa. In particular, a subset of high-titer inhibitors responded to high dose fVIII, which would not be predicted based on their inhibitor titer alone. Thus the ability to quickly map the epitope spectrum of patient plasma using a clinically feasible assay may fundamentally change how clinicians approach the treatment of high-titer inhibitor patients. Objectives To map the epitopes of anti-fVIII MAbs, of which 3 are classical inhibitors and one non-classical, using hydrogen-deuterium exchange coupled with liquid chromatography-mass spectrometry (HDX-MS). Methods Binding epitopes of 4 MAbs targeting fVIII C2 domain were mapped using HDX-MS. Results The epitopes determined by HDX-MS are consistent with those obtained earlier through structural characterization and antibody competition assays. In addition classical and non-classical inhibitor epitopes could be distinguished using a limited subset of C2-derived peptic fragments. Conclusion Our results demonstrate the effectiveness and robustness of the HDX-MS method for epitope mapping and suggest a potential role of rapid mapping of fVIII inhibitor epitopes in facilitating individualized treatment of inhibitor patients. PMID:24152306

  6. Understanding the conformational impact of chemical modifications on monoclonal antibodies with diverse sequence variation using hydrogen/deuterium exchange mass spectrometry and structural modeling.

    Science.gov (United States)

    Zhang, Aming; Hu, Ping; MacGregor, Paul; Xue, Yu; Fan, Haihong; Suchecki, Peter; Olszewski, Leonard; Liu, Aston

    2014-04-01

    Chemical modifications can potentially induce conformational changes near the modification site and thereby impact the safety and efficacy of protein therapeutics. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) has emerged as a powerful analytical technique with high spatial resolution and sensitivity in detecting such local conformational changes. In this study, we utilized HDX-MS combined with structural modeling to examine the conformational impact on monoclonal antibodies (mAbs) caused by common chemical modifications including methionine (Met) oxidation, aspartic acid (Asp) isomerization, and asparagine (Asn) deamidation. Four mAbs with diverse sequences and glycosylation states were selected. The data suggested that the impact of Met oxidation was highly dependent on its location and glycosylation state. For mAbs with normal glycosylation in the Fc region, oxidation of the two conserved Met252 and Met428 (Kabat numbering) disrupted the interface interactions between the CH2 and CH3 domains, thus leading to a significant decrease in CH2 domain thermal stability as well as a slight increase in aggregation propensity. In contrast, Met oxidation in the variable region and CH3 domain had no detectable impact on mAb conformation. For aglycosylated mAb, Met oxidation could cause a more global conformational change to the whole CH2 domain, coincident with the larger decrease in thermal stability and significant increase in aggregation rate. Unlike Met oxidation, Asn deamidation and Asp isomerization mostly had very limited effects on mAb conformation, with the exception of succiminide intermediate formation which induced a measurable local conformational change to be more solvent protected. Structural modeling suggested that the succinimide intermediate was stabilized by adjacent aromatic amino acids through ring-ring stacking interactions.

  7. Epitope mapping of inhibitory antibodies targeting the C2 domain of coagulation factor VIII by hydrogen-deuterium exchange mass spectrometry.

    Science.gov (United States)

    Sevy, A M; Healey, J F; Deng, W; Spiegel, P C; Meeks, S L; Li, R

    2013-12-01

    The development of anti-factor VIII antibodies (inhibitors) is a significant complication in the management of patients with hemophilia A, leading to significant increases in morbidity and treatment cost. Using a panel of mAbs against different epitopes on FVIII, we have recently shown that epitope specificity, inhibitor kinetics and time to maximum inhibition are more important than inhibitor titer in predicting responses to FVIII and the combination of FVIII and recombinant FVIIa. In particular, a subset of high-titer inhibitors responded to high-dose FVIII, which would not be predicted on the basis of their inhibitor titer alone. Thus, the ability to quickly map the epitope spectrum of patient plasma with a clinically feasible assay may fundamentally change how clinicians approach the treatment of high-titer inhibitor patients. To map the epitopes of anti-FVIII mAbs, three of which are classic inhibitors and one of which is a non-classic inhibitor, by the use of hydrogen-deuterium exchange coupled with mass spectrometry (HDX-MS). The binding epitopes of four mAbs targeting the FVIII C2 domain were mapped with HDX-MS. The epitopes determined with HDX-MS are consistent with those obtained earlier through structural characterization and antibody competition assays. In addition, classic and non-classic inhibitor epitopes could be distinguished by the use of a limited subset of C2 domain-derived peptic fragments. Our results demonstrate the effectiveness and robustness of the HDX-MS method for epitope mapping, and suggest a potential role of rapid mapping of FVIII inhibitor epitopes in facilitating individualized treatment of inhibitor patients. © 2013 International Society on Thrombosis and Haemostasis.

  8. Structural Dynamics of the GW182 Silencing Domain Including its RNA Recognition motif (RRM) Revealed by Hydrogen-Deuterium Exchange Mass Spectrometry

    Science.gov (United States)

    Cieplak-Rotowska, Maja K.; Tarnowski, Krzysztof; Rubin, Marcin; Fabian, Marc R.; Sonenberg, Nahum; Dadlez, Michal; Niedzwiecka, Anna

    2018-01-01

    The human GW182 protein plays an essential role in micro(mi)RNA-dependent gene silencing. miRNA silencing is mediated, in part, by a GW182 C-terminal region called the silencing domain, which interacts with the poly(A) binding protein and the CCR4-NOT deadenylase complex to repress protein synthesis. Structural studies of this GW182 fragment are challenging due to its predicted intrinsically disordered character, except for its RRM domain. However, detailed insights into the properties of proteins containing disordered regions can be provided by hydrogen-deuterium exchange mass spectrometry (HDX/MS). In this work, we applied HDX/MS to define the structural state of the GW182 silencing domain. HDX/MS analysis revealed that this domain is clearly divided into a natively unstructured part, including the CCR4-NOT interacting motif 1, and a distinct RRM domain. The GW182 RRM has a very dynamic structure, since water molecules can penetrate the whole domain in 2 h. The finding of this high structural dynamics sheds new light on the RRM structure. Though this domain is one of the most frequently occurring canonical protein domains in eukaryotes, these results are - to our knowledge - the first HDX/MS characteristics of an RRM. The HDX/MS studies show also that the α2 helix of the RRM can display EX1 behavior after a freezing-thawing cycle. This means that the RRM structure is sensitive to environmental conditions and can change its conformation, which suggests that the state of the RRM containing proteins should be checked by HDX/MS in regard of the conformational uniformity. [Figure not available: see fulltext.

  9. A biological method of including mineralized human liquid and solid wastes into the mass exchange of bio-technical life support systems

    Science.gov (United States)

    Ushakova, S. A.; Tikhomirov, A. A.; Tikhomirova, N. A.; Kudenko, Yu. A.; Litovka, Yu. A.; Anishchenko, O. V.

    2012-10-01

    The main obstacle to using mineralized human solid and liquid wastes as a source of mineral elements for plants cultivated in bio-technical life support systems (BLSS) is that they contain NaCl. The purpose of this study is to determine whether mineralized human wastes can be used to prepare the nutrient solution for long-duration conveyor cultivation of uneven-aged wheat and Salicornia europaea L. plant community. Human solid and liquid wastes were mineralized by the method of "wet incineration" developed by Yu. Kudenko. They served as a basis for preparing the solutions that were used for conveyor-type cultivation of wheat community represented by 5 age groups, planted with a time interval of 14 days. Wheat was cultivated hydroponically on expanded clay particles. To reduce salt content of the nutrient solution, every two weeks, after wheat was harvested, 12 L of solution was removed from the wheat irrigation tank and used for Salicornia europaea cultivation in water culture in a conveyor mode. The Salicornia community was represented by 2 age groups, planted with a time interval of 14 days. As some portion of the nutrient solution used for wheat cultivation was regularly removed, sodium concentration in the wheat irrigation solution did not exceed 400 mg/L, and mineral elements contained in the removed portion were used for Salicornia cultivation. The experiment lasted 4 months. The total wheat biomass productivity averaged 30.1 g · m-2 · day-1, and the harvest index amounted to 36.8%. The average productivity of Salicornia edible biomass on a dry weight basis was 39.3 g · m-2 · day-1, and its aboveground mass contained at least 20% of NaCl. Thus, the proposed technology of cultivation of wheat and halophyte plant community enables using mineralized human wastes as a basis for preparing nutrient solutions and including NaCl in the mass exchange of the BLSS; moreover, humans are supplied with additional amounts of leafy vegetables.

  10. Rapid isolation of plutonium in environmental solid samples using sequential injection anion exchange chromatography followed by detection with inductively coupled plasma mass spectrometry

    DEFF Research Database (Denmark)

    Qiao, Jixin; Hou, Xiaolin; Roos, Per

    2011-01-01

    is straightforward and less labor intensive as compared with batch-wise anion exchange chromatographic procedures. Besides, the automated method features low consumption of ion-exchanger and reagents for column washing and elution, with the consequent decrease in the generation of acidic waste, thus bearing green...

  11. Barter exchanges

    DEFF Research Database (Denmark)

    Sudzina, Frantisek

    Although barter is often perceived as something that proceeded money, barter is still used. The focus of the paper is on barter exchanges. Barter exchanges are used both in developing countries as well as in developed countries (including the U.S.). They are used by both organizations...... and individuals. They usually allow to exchange good but some include also services. Some exchanges allow only for bi-directional barter, i.e. when only two parties are involved in the exchange. But probably most of the barter exchanges use barter money; this makes it easier to exchange goods and services...

  12. Decentralized SINR Balancing in Cognitive Radio Networks

    KAUST Repository

    Dhifallah, Oussama Najeeb

    2016-07-07

    This paper considers the downlink of a cognitive radio (CR) network formed by multiple primary and secondary transmitters, where each multi-antenna transmitter serves a pre-known set of single-antenna users. The paper assumes that the secondary and primary transmitters can transmit simultaneously their data over the same frequency bands, so as to achieve a high system spectrum efficiency. The paper considers the downlink balancing problem of maximizing the minimum signal-to-interference-plus noise ratio (SINR) of the secondary transmitters subject to both total power constraint of the secondary transmitters, and maximum interference constraint at each primary user due to secondary transmissions. The paper proposes solving the problem using the alternating direction method of multipliers (ADMM), which leads to a distributed implementation through limited information exchange across the coupled secondary transmitters. The paper additionally proposes a solution that guarantees feasibility at each iteration. Simulation results demonstrate that the proposed solution converges to the centralized solution in a reasonable number of iterations.

  13. Simple setup for gas-phase h/d exchange mass spectrometry coupled to electron transfer dissociation and ion mobility for analysis of polypeptide structure on a liquid chromatographic time scale

    DEFF Research Database (Denmark)

    Mistarz, Ulrik Hvid; Brown, Jeffery M; Haselmann, Kim F

    2014-01-01

    Gas-phase hydrogen/deuterium exchange (HDX) is a fast and sensitive, yet unharnessed analytical approach for providing information on the structural properties of biomolecules, in a complementary manner to mass analysis. Here, we describe a simple setup for ND3-mediated millisecond gas-phase HDX...... gas immediately upstream or downstream of the primary skimmer cone. The approach was implemented on three commercially available mass spectrometers and required no or minor fully reversible reconfiguration of gas-inlets of the ion source. Results from gas-phase HDX-MS of peptides using the aqueous ND3...

  14. Co-existence of two different α-synuclein oligomers with different core structures determined by hydrogen/deuterium exchange mass spectrometry

    DEFF Research Database (Denmark)

    Paslawski, Wojciech; Mysling, Simon; Thomsen, Karen

    2014-01-01

    Neurodegenerative disorders are characterized by the formation of protein oligomers and amyloid fibrils, which in the case of Parkinson's disease involves the protein α-synuclein (αSN). Cytotoxicity is mainly associated with the oligomeric species, but we still know little about their assembly...... are protected from exchange with D2 O until they dissociate into monomeric αSN by EX1 exchange kinetics. Fewer residues are protected against exchange in oligomer type II, but this type does not revert to αSN monomers. Both oligomers are protected in the core sequence Y39-A89. Based on incubation studies...

  15. Study on autonomous decentralized-cooperative function monitoring system

    International Nuclear Information System (INIS)

    Matsuoka, Takeshi; Numano, Masayoshi; Someya, Minoru; Fukuto, Junji; Mitomo, Nobuo; Miyazaki, Keiko; Matsukura, Hiroshi; Tanba, Yasuyuki

    1999-01-01

    In this study, a study further advanced on a base of results of study on artificial intelligence for nuclear power', one of nuclear basis crossover studies, conducted at five years planning from 1989 fiscal year was executed. Here was conducted on study on a system technology for supplying cooperation, judgement process, judgement results, and so forth between decentralized artificial intelligent elements (agents) to operation managers (supervisors) by focussing a system for monitoring if autonomous decentralized system containing plant operation and robot group action functioned appropriately. In 1997 fiscal year, by mainly conducting development for displaying working state of robot group, some investigations on integrated management of each function already development and maintained were executed. Furthermore, some periodical meetings on realization of its integration with operation control system and maintenance system with other research institutes were conducted. (G.K.)

  16. Decentralized control mechanism underlying interlimb coordination of millipedes.

    Science.gov (United States)

    Kano, Takeshi; Sakai, Kazuhiko; Yasui, Kotaro; Owaki, Dai; Ishiguro, Akio

    2017-04-04

    Legged animals exhibit adaptive and resilient locomotion through interlimb coordination. The long-term goal of this study is to clarify the relationship between the number of legs and the inherent decentralized control mechanism for interlimb coordination. As a preliminary step, the study focuses on millipedes as they represent the species with the greatest number of legs among various animal species. A decentralized control mechanism involving local force feedback was proposed based on the qualitative findings of behavioural experiments in which responses to the removal of part of the terrain and leg amputation were observed. The proposed mechanism was implemented in a developed millipede-like robot to demonstrate that the robot can adapt to the removal of the part of the terrain and leg amputation in a manner similar to that in behavioural experiments.

  17. Coordination in the Decentralized Assembly System with Dual Supply Modes

    Directory of Open Access Journals (Sweden)

    Xu Guan

    2013-01-01

    Full Text Available This paper investigates a decentralized assembly system that consists of one assembler and two independent suppliers; wherein one supplier is perfectly reliable for the production, while the other generates yield uncertainty. Facing the random market demand, the assembler has to order the components from one supplier in advance and meanwhile requires the other supplier to deliver the components under VMI mode. We construct a Nash game between the supplier and the assembler so as to derive their equilibrium procurement/production strategies. The results show that the channel’s performance is highly undermined by the decentralization between players and also the combination of two supply modes. Compared to the centralized system, we propose an advance payment contract to perfectly coordinate the supply chain performance. The numerical examples indicate some management implications on the supply mode comparison and sensitivity analysis.

  18. [Decentralization of health services: case studies in six Mexican states].

    Science.gov (United States)

    Homedes, Nuria; Ugalde, Antonio

    2011-01-01

    This article discusses the effects of the second decentralization of the Mexican Ministry of Health (1994-2000). It is based on a review of official and archival documents, health and productivity statistics, observations in clinics and hospitals, and 232 in-depth interviews in the states of Baja California Sur, Colima, Guanajuato, Nuevo León, Sonora and Tamaulipas. The interviewees included high-level administrators of state and district health systems, leaders of workers unions, health providers and representatives of civil society. The article identifies the problems that offices of health at state level had to overcome to implement the decentralization. Descentralizacion failed to achieve the objectives stated by its promoters.

  19. Model-Based Design of Tree WSNs for Decentralized Detection

    Directory of Open Access Journals (Sweden)

    Ashraf Tantawy

    2015-08-01

    Full Text Available The classical decentralized detection problem of finding the optimal decision rules at the sensor and fusion center, as well as variants that introduce physical channel impairments have been studied extensively in the literature. The deployment of WSNs in decentralized detection applications brings new challenges to the field. Protocols for different communication layers have to be co-designed to optimize the detection performance. In this paper, we consider the communication network design problem for a tree WSN. We pursue a system-level approach where a complete model for the system is developed that captures the interactions between different layers, as well as different sensor quality measures. For network optimization, we propose a hierarchical optimization algorithm that lends itself to the tree structure, requiring only local network information. The proposed design approach shows superior performance over several contentionless and contention-based network design approaches.

  20. Decentralized Neural Backstepping Control Applied to a Robot Manipulator

    Directory of Open Access Journals (Sweden)

    Ramon Garcia-Hernandez

    2013-01-01

    Full Text Available This paper presents a discrete-time decentralized control scheme for trajectory tracking of a two degrees of freedom (DOF robot manipulator. A high order neural network (HONN is used to approximate a decentralized control law designed by the backstepping technique as applied to a block strict feedback form (BSFF. The weights for each neural network are adapted online by an extended Kalman filter training algorithm. The motion for each joint is controlled independently using only local angular position and velocity measurements. The stability analysis for the closed-loop system via the Lyapunov approach is included. Finally, the real-time results show the feasibility of the proposed control scheme using a robot manipulator.

  1. Decentralized cogeneration - A solution for Romania? RAEF experience

    International Nuclear Information System (INIS)

    Binig, Alexandru-Valeriu

    2004-01-01

    deficit should be covered using indigenous power generation sources. One could conclude, correlated with the previous discussion on financing investment, that most of the private capital is expected in the power and heat generation sector. New injection points might necessitate network reinforcement (implying additional costs and delays), may have to pass a complicated and lengthy authorizations process, etc. In conclusion, realisation of 'greenfield' large projects is likely to be cumbersome and delayed due mainly to issues related to connection to the grid. But also fuel supply issues, cooling water access, impact on communities, add to the serious siting problems for new greenfield power generation projects. A psychological impact on private investor's appetite is also given by the continuation of erection of Cernavoda 2, (3?) nuclear units, as these are modern, safe, and optimal at dispatching. In conclusion, in Romania, at present, one cannot identify large private power generation projects in advanced development phase. A solution could be decentralized power generation. Combined with covering a heat demand, it leads to distributed cogeneration. It is an EU and worldwide trend. The above, combined with the overall analysis of the experience in Romania and worldwide allow drawing the following conclusions: - Decentralised generation (cogeneration) is a solution to be considered for Romania and for the region; - The national energy strategy must consider it; - Resources should be allocated to it (not necessarily financial), thus contributing to sustainable development; - It must be promoted by appropriate legislative, regulatory fiscal, etc framework; - Periodic exchange of experience among different actors is decisive for avoiding wasting resources; - this is mainly the purpose of the present article; - Private initiative is the main driver and must be encouraged for promoting efficiency and sustainability; - The solution chosen yields better risk management while

  2. A peer-to-peer platform for decentralized logistics

    OpenAIRE

    Gallay, Olivier; Korpela, Kari; Tapio, Niemi; Nurminen, Jukka K.; Kersten, Wolfgang; Blecker, Thorsten; Ringle, Christian M.

    2017-01-01

    We introduce a novel platform for decentralized logistics, the aim of which is to magnify and accelerate the impact offered by the integration of the most recent advances in Information and Communication Technologies (ICTs) to multi-modal freight operations. The essence of our peer-to-peer (P2P) framework distributes the management of the logistics operations to the multiple actors according to their available computational resources. As a result, this new approach prevents the dominant playe...

  3. Subsecond Tsunamis and Delays in Decentralized Electronic Systems

    Directory of Open Access Journals (Sweden)

    Pedro D. Manrique

    2017-10-01

    Full Text Available Driven by technological advances and economic gain, society’s electronic systems are becoming larger, faster, more decentralized and autonomous, and yet with increasing global reach. A prime example are the networks of financial markets which—in contrast to popular perception—are largely all-electronic and decentralized with no top-down real-time controller. This prototypical system generates complex subsecond dynamics that emerge from a decentralized network comprising heterogeneous hardware and software components, communications links, and a diverse ecology of trading algorithms that operate and compete within this all-electronics environment. Indeed, these same technological and economic drivers are likely to generate a similarly competitive all-electronic ecology in a variety of future cyberphysical domains such as e-commerce, defense and the transportation system, including the likely appearance of large numbers of autonomous vehicles on the streets of many cities. Hence there is an urgent need to deepen our understanding of stability, safety and security across a wide range of ultrafast, large, decentralized all-electronic systems—in short, society will eventually need to understand what extreme behaviors can occur, why, and what might be the impact of both intentional and unintentional system perturbations. Here we set out a framework for addressing this issue, using a generic model of heterogeneous, adaptive, autonomous components where each has a realistic limit on the amount of information and processing power available to it. We focus on the specific impact of delayed information, possibly through an accidental shift in the latency of information transmission, or an intentional attack from the outside. While much remains to be done in terms of developing formal mathematical results for this system, our preliminary results indicate the type of impact that can occur and the structure of a mathematical theory which may

  4. Centralized versus decentralized taxation of mobile polluting firms

    International Nuclear Information System (INIS)

    Tanguay, Georges A.; Marceau, Nicolas

    2001-01-01

    We consider a world in which a mobile polluting firm must locate in one of two regions. The regions differ in two dimensions: their marginal cost of pollution and the production cost of the firm. It is shown that under incomplete information on regional marginal costs of pollution, fiscal competition may lead to the sub-optimal location of the firm. We also show that under incomplete information, a sub-optimal location is less likely under centralized than under decentralized taxation

  5. Impacts of Decentralization on Environmental Management in Thailand

    Directory of Open Access Journals (Sweden)

    Soparatana Jarusombat

    2010-08-01

    Full Text Available This paper describes the process, institutional and legal framework within which the environmental management operates in Thailand. It specifically focuses on the decentralization within central and local government’s role in environmental management. The methods of this research use literature review. The aim of the paper is to examine how interface between the central and local loci of power have affected pieces of legislation relating to management of the environment by central and local government in Thailand.

  6. Dynamical Orders of Decentralized H-infinity Controllers

    DEFF Research Database (Denmark)

    Stoustrup, Jakob; Niemann, Hans Henrik

    1996-01-01

    The problem of decentralized control is addressed, i.e. theproblem of designing a controller where each control input is allowedto use only some of the measurements. It is shown that such problemsthere does not always exist a sequence of controllers of bounded orderwhich obtains near optimal cont...... control. Neither does there exist an infinitedimensional optimal controller. Using the insight of the line of proofof these results, a heuristic design algorithm is proposed for designingnear optimal controllers of increasing orders....

  7. Decentralized energy-saving hydraulic concepts for mobile working machines

    OpenAIRE

    Lodewyks, Johann; Zurbrügg, Pascal

    2016-01-01

    The high price of batteries in working machines with electric drives offer a potential for investment in energy-saving hydraulic systems. The decentralized power network opens up new approaches for hydraulic- and hybrid circuits. In addition, the regeneration of energy can be used at any point of the machine. For the example of an excavator arm drive with a double cylinder two compact hydraulic circuits are presented, which relieve a central hydraulic system.

  8. Blockchain technology and decentralized governance: Is the state still necessary?

    OpenAIRE

    Marcella Atzori

    2017-01-01

    The core technology of Bitcoin, the blockchain, has recently emerged as a disruptive innovation with a wide range of applications, potentially able to redesign our interactions in business, politics and society at large. Although scholarly interest in this subject is growing, a comprehensive analysis of blockchain applications from a political perspective is severely lacking to date. This paper aims to fill this gap and it discusses the key points of blockchain-based decentralized governance,...

  9. The commercialization of decentralized energy systems (DES) in the Philippines

    International Nuclear Information System (INIS)

    Bernardo, J.Y.; Navarro, L.B.; Abito, G.; Lim, B.P.

    1992-01-01

    PNOC-ERDC is implementing the project ''Commercialization of Decentralized Energy Systems or DES'' with assistance from the European Community. The project hopes to promote the utilization of DES technologies by providing financial and technical assistance to enterpreneurs engaged in the manufacture and/or distribution of DES technologies. At present, the DES project has provided loans totalling P17.9 million to 6 entrepreneurs. Technologies supported include photovoltaics, biogas and agricultural waste-fired dryers. (auth.). 2 tabs

  10. Blockchain technology and decentralized governance: Is the state still necessary?

    Directory of Open Access Journals (Sweden)

    Marcella Atzori

    2017-03-01

    Full Text Available The core technology of Bitcoin, the blockchain, has recently emerged as a disruptive innovation with a wide range of applications, potentially able to redesign our interactions in business, politics and society at large. Although scholarly interest in this subject is growing, a comprehensive analysis of blockchain applications from a political perspective is severely lacking to date. This paper aims to fill this gap and it discusses the key points of blockchain-based decentralized governance, which challenges to varying degrees the traditional mechanisms of State authority, citizenship and democracy. In particular, the paper verifies to which extent blockchain and decentralized platforms can be considered as hyper-political tools, capable to manage social interactions on large scale and dismiss traditional central authorities. The analysis highlights risks related to a dominant position of private powers in distributed ecosystems, which may lead to a general disempowerment of citizens and to the emergence of a stateless global society. While technological utopians urge the demise of any centralized institution, this paper advocates the role of the State as a necessary central point of coordination in society, showing that decentralization through algorithm-based consensus is an organizational theory, not a stand-alone political theory.

  11. Robust Decentralized Nonlinear Control for a Twin Rotor MIMO System.

    Science.gov (United States)

    Belmonte, Lidia María; Morales, Rafael; Fernández-Caballero, Antonio; Somolinos, José Andrés

    2016-07-27

    This article presents the design of a novel decentralized nonlinear multivariate control scheme for an underactuated, nonlinear and multivariate laboratory helicopter denominated the twin rotor MIMO system (TRMS). The TRMS is characterized by a coupling effect between rotor dynamics and the body of the model, which is due to the action-reaction principle originated in the acceleration and deceleration of the motor-propeller groups. The proposed controller is composed of two nested loops that are utilized to achieve stabilization and precise trajectory tracking tasks for the controlled position of the generalized coordinates of the TRMS. The nonlinear internal loop is used to control the electrical dynamics of the platform, and the nonlinear external loop allows the platform to be perfectly stabilized and positioned in space. Finally, we illustrate the theoretical control developments with a set of experiments in order to verify the effectiveness of the proposed nonlinear decentralized feedback controller, in which a comparative study with other controllers is performed, illustrating the excellent performance of the proposed robust decentralized control scheme in both stabilization and trajectory tracking tasks.

  12. Decentralization Challenges for Management of Cultural Patrimony in Ecuador

    Directory of Open Access Journals (Sweden)

    Dr.C. Carlos Leonel Escudero-Sánchez

    2015-10-01

    Full Text Available In Latin America  the new  decentralization policies in  the forms of government  challenging  institutional  processes  and management  practices.  In the context of  Ecuador  responds to  a constitutional  mandate expressed in  the reformulation of  the powers of the  municipal  autonomous governments. Consequently, the main  purpose of the article  is to present  the main  principles  expressed  in the  protection, evaluation and dissemination of cultural heritage.  Therefore  the  exercise of autonomy  and decentralization  is governed  by the principles  of solidarity, subsidiarity,  territorial equity, integration and participation.  Hence,  the main results  are  part of  the systematization of  such  socio-economic, institutional, governance  and participation, legal and financial  management of cultural heritage contexts.Keywords: decentralization, cultural heritage, cultural management, citizen participation.

  13. Game-Based Virtual Worlds as Decentralized Virtual Activity Systems

    Science.gov (United States)

    Scacchi, Walt

    There is widespread interest in the development and use of decentralized systems and virtual world environments as possible new places for engaging in collaborative work activities. Similarly, there is widespread interest in stimulating new technological innovations that enable people to come together through social networking, file/media sharing, and networked multi-player computer game play. A decentralized virtual activity system (DVAS) is a networked computer supported work/play system whose elements and social activities can be both virtual and decentralized (Scacchi et al. 2008b). Massively multi-player online games (MMOGs) such as World of Warcraft and online virtual worlds such as Second Life are each popular examples of a DVAS. Furthermore, these systems are beginning to be used for research, deve-lopment, and education activities in different science, technology, and engineering domains (Bainbridge 2007, Bohannon et al. 2009; Rieber 2005; Scacchi and Adams 2007; Shaffer 2006), which are also of interest here. This chapter explores two case studies of DVASs developed at the University of California at Irvine that employ game-based virtual worlds to support collaborative work/play activities in different settings. The settings include those that model and simulate practical or imaginative physical worlds in different domains of science, technology, or engineering through alternative virtual worlds where players/workers engage in different kinds of quests or quest-like workflows (Jakobsson 2006).

  14. Decentralization, healthcare access, and inequality in Mpumalanga, South Africa.

    Science.gov (United States)

    Winchester, Margaret S; King, Brian

    2018-04-27

    Healthcare access and utilization remain key challenges in the Global South. South Africa represents this given that more than twenty years after the advent of democratic elections, the national government continues to confront historical systems of spatial manipulation that generated inequities in healthcare access. While the country has made significant advancements, governmental agencies have mirrored international strategies of healthcare decentralization and focused on local provision of primary care to increase healthcare access. In this paper, we show the significance of place in shaping access and health experiences for rural populations. Using data from a structured household survey, focus group discussions, qualitative interviews, and clinic data conducted in northeast South Africa from 2013 to 2016, we argue that decentralization fails to resolve the uneven landscapes of healthcare in the contemporary period. This is evidenced by the continued variability across the study area in terms of government-sponsored healthcare, and constraints in the clinics in terms of staffing, privacy, and patient loads, all of which challenge the access-related assumptions of healthcare decentralization. Copyright © 2018 Elsevier Ltd. All rights reserved.

  15. The Financing of the Costs of Education under Decentralization

    Directory of Open Access Journals (Sweden)

    Kovalenko Svitlana O.

    2017-08-01

    Full Text Available The article is aimed at determining the major problems in the financing of education sphere in the current context of the reform of decentralization of power in Ukraine and developing directions for solving them. The main directions and results of educational reform have been analyzed, and the ranking of Ukraine according to the Global Competitiveness Index has been defined. The positive points of the decentralization of education have been noted. The expenditure of the budget of Ukraine have been analyzed and the proportion of expenditure on education has been determined as to the total budget expenditure. Along with this, the main circle of problems of the budget provision for the education sector has been outlined. It has been proposed: to increase the efficiency of allocation of budgetary means at all levels of the budget system; to enhance the possibilities of educational institutions to attract additional financial resources through educational innovation, and to expand the list of paid services provided by such institutions; to decentralize education, taking into account the characteristics of development of regions, their productive potential, the demographic situation, and the interests of the territorial communities.

  16. Different conformational dynamics of β-arrestin1 and β-arrestin2 analyzed by hydrogen/deuterium exchange mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Yun, Youngjoo; Kim, Dong Kyun [School of Pharmacy, Sungkyunkwan University, Suwon (Korea, Republic of); Seo, Min-Duk [College of Pharmacy & Department of Molecular Science and Technology, Ajou University, Suwon (Korea, Republic of); Kim, Kyeong-Man [College of Pharmacy, Chonnam National University, Gwang-Ju (Korea, Republic of); Chung, Ka Young, E-mail: kychung2@skku.edu [School of Pharmacy, Sungkyunkwan University, Suwon (Korea, Republic of)

    2015-01-30

    Highlights: • The conformational dynamics of β-arrestin1 or β-arrestin2 were analyzed by HDX-MS. • β-Strands II through IV were more dynamic in β-arrestin2 than in β-arrestin1. • The middle loop was less dynamic in β-arrestin2 than in β-arrestin1. • Upon pre-activation by the R169E mutation, β-arrestins became more dynamic. • Pre-activation affected a wider region of β-arrestin1 compared to β-arrestin2. - Abstract: Arrestins have important roles in G protein-coupled receptor (GPCR) signaling including desensitization of GPCRs and G protein-independent signaling. There have been four arrestins identified: arrestin1, arrestin2 (e.g. β-arrestin1), arrestin3 (e.g. β-arrestin2), and arrestin4. β-Arrestin1 and β-arrestin2 are ubiquitously expressed and regulate a broad range of GPCRs, while arrestin1 and arrestin4 are expressed in the visual system. Although the functions of β-arrestin1 and β-arrestin2 widely overlap, β-arrestin2 has broader receptor selectivity, and a few studies have suggested that β-arrestin1 and β-arrestin2 have distinct cellular functions. Here, we compared the conformational dynamics of β-arrestin1 and β-arrestin2 by hydrogen/deuterium exchange mass spectrometry (HDX-MS). We also used the R169E mutant as a pre-activation model system. HDX-MS data revealed that β-strands II through IV were more dynamic in β-arrestin2 in the basal state, while the middle loop was more dynamic in β-arrestin1. With pre-activation, both β-arrestin1 and β-arrestin2 became more flexible, but broader regions of β-arrestin1 became flexible compared to β-arrestin2. The conformational differences between β-arrestin1 and β-arrestin2 in both the basal and pre-activated states might determine their different receptor selectivities and different cellular functions.

  17. Bulk derivatization and cation exchange restricted access media-based trap-and-elute liquid chromatography–mass spectrometry method for determination of trace estrogens in serum

    Energy Technology Data Exchange (ETDEWEB)

    Beinhauer, Jana [Department of Biochemistry, Faculty of Science, Palacký University, Šlechtitelů 11, CZ-783 71 Olomouc (Czech Republic); Centre of the Region Haná for Biotechnological and Agricultural Research - Department of Protein Biochemistry and Proteomics, Faculty of Science, Palacký University, Šlechtitelů 11, CZ-783 71 Olomouc (Czech Republic); Bian, Liangqiao [Shimadzu Center for Advanced Analytical Chemistry, The University of Texas at Arlington, Arlington, TX (United States); Shimadzu Institute for Research Technologies, The University of Texas at Arlington, Arlington, TX (United States); Fan, Hui [Department of Chemistry and Biochemistry, The University of Texas at Arlington, Arlington, TX (United States); Šebela, Marek [Department of Biochemistry, Faculty of Science, Palacký University, Šlechtitelů 11, CZ-783 71 Olomouc (Czech Republic); Centre of the Region Haná for Biotechnological and Agricultural Research - Department of Protein Biochemistry and Proteomics, Faculty of Science, Palacký University, Šlechtitelů 11, CZ-783 71 Olomouc (Czech Republic); Kukula, Maciej [Shimadzu Center for Advanced Analytical Chemistry, The University of Texas at Arlington, Arlington, TX (United States); Shimadzu Institute for Research Technologies, The University of Texas at Arlington, Arlington, TX (United States); Barrera, Jose A. [Shimadzu Institute for Research Technologies, The University of Texas at Arlington, Arlington, TX (United States); and others

    2015-02-09

    Highlights: • Analysis of estrogens in small volume samples at low parts-per-trillion concentration. • Charged bulk derivatization facilitates on-line ion exchange sample preparation. • On-line WCX restricted access media traps analytes, but not proteins and lipids. • Complete preparation and LC–MS/MS analysis completed in 30 min/sample. - Abstract: Estrone (E1), estradiols (α/β-E2), and estriol (E3) are four major metabolically active estrogens exerting strong biological activities at very low circulating concentrations. This paper reports a sensitive and efficient method with automated, on-line clean-up and detection to determine trace estrogens in a small volume of serum samples using liquid chromatography–electrospray ionization–tandem mass spectrometry directly, without off-line liquid–liquid or solid-phase extraction pretreatments. Serum aliquots (charcoal stripped fetal bovine serum, 100 μL) were spiked with four estrogen standards and their corresponding isotope-labeled internal standards, then bulk derivatized with 2-fluoro-1-methyl-pyridium p-toluenesulfonate (2-FMP) to establish the calibration curves and perform method validation. Calibration was established in the concentration ranges of 5–1000 pg mL{sup −1}, and demonstrated good linearity of R{sup 2} from 0.9944 to 0.9997 for the four derivatized estrogens. The lower detection limits obtained were 3–7 pg mL{sup −1}. Good accuracy and precision in the range of 86–112% and 2.3–11.9%, respectively, were observed for the quality control (QC) samples at low, medium, and high concentration levels. The stability tests showed that the derivatized serum samples were stable 8 h after derivatization at room temperature and at least to 48 h if stored at −20 °C. The method was applied to measure trace estrogens in real human and bovine serum samples, and three of four estrogen compounds studied were observed and quantified.

  18. Decentralized, cooperative control of multivehicle systems: Design and stability analysis

    Science.gov (United States)

    Weitz, Lesley Anne

    2009-12-01

    This dissertation addresses the design and stability analysis of decentralized, cooperative control laws for multivehicle systems. Advances in communication, navigation, and surveillance systems have enabled greater autonomy in multivehicle systems, and there is a shift toward decentralized, cooperative systems for computational efficiency and robustness. In a decentralized control scheme, control inputs are determined onboard each vehicle; therefore, decentralized controllers are more efficient for large numbers of vehicles, and the system is more robust to communication failures and reconfiguration. The design of decentralized, cooperative control laws is explored for a nonlinear vehicle model that can be represented in a double-integrator form. Cooperative controllers are functions of spacing errors with respect to other vehicles in the system, where the communication structure defines the information that is available to each vehicle. Control inputs are selected to achieve internal stability, or zero steady-state spacing errors, between vehicles in the system. Closed-loop equations of motion for the cooperative system can be written in a structural form, where damping and stiffness matrices contain control gains acting on the velocity and positions of the vehicles, respectively. The form of the stiffness matrix is determined by the communication structure, where different communication structures yield different control forms. Communication structures are compared using two structural analysis tools: modal cost and frequency-response functions, which evaluate the response of the multivehicle systems to disturbances. The frequency-response information is shown to reveal the string stability of different cooperative control forms. The effects of time delays in the feedback states of the cooperative control laws on system stability are also investigated. Closed-loop equations of motion are modeled as delay differential equations, and two stability notions are

  19. Forms of government decentralization and institutional quality: Evidence from a large sample of nations

    OpenAIRE

    Goel, Rajeev K.; Saunoris, James W.

    2016-01-01

    This paper studies the effects of various forms of government decentralization on institutional quality across countries. Using corruption and the shadow economy to proxy for institutional quality, as well as three forms of government decentralization (i.e., virtual, physical, and fiscal), the econometric results show virtual decentralization to be the most effective in improving institutional quality. The effects on transition and countries in Asia are also considered.

  20. (De)Centralization of Public Procurement at the Local Level in the EU

    OpenAIRE

    Boštjan BREZOVNIK; Žan Jan OPLOTNIK; Borut VOJINOVIĆ

    2015-01-01

    The so-called decentralization of public procurement in EU Member States is accepted as the most suitable design of the public procurement system, often justifi ed by greater economic effi ciency and by the possibility of boosting the development of small and medium-sized enterprises, which act on the public procurement market as providers of goods, services and works. Despite the existence of highly decentralized public procurement systems which refl ect the decentralization of administrativ...

  1. A Mixed Method Research for Finding a Model of Administrative Decentralization

    OpenAIRE

    Tahereh Feizy; Alireza Moghali; Masuod Geramipoor; Reza Zare

    2015-01-01

    One of the critical issues of administrative decentralization in translating theory into practice is understanding its meaning. An important method to identify administrative decentralization is to address how it can be planned and implemented, and what are its implications, and how it would overcome challenges. The purpose of this study is finding a model for analyzing and evaluating administrative decentralization, so a mixed method research was used to explore and confirm the model of Admi...

  2. Fuzzy Adaptive Decentralized Optimal Control for Strict Feedback Nonlinear Large-Scale Systems.

    Science.gov (United States)

    Sun, Kangkang; Sui, Shuai; Tong, Shaocheng

    2018-04-01

    This paper considers the optimal decentralized fuzzy adaptive control design problem for a class of interconnected large-scale nonlinear systems in strict feedback form and with unknown nonlinear functions. The fuzzy logic systems are introduced to learn the unknown dynamics and cost functions, respectively, and a state estimator is developed. By applying the state estimator and the backstepping recursive design algorithm, a decentralized feedforward controller is established. By using the backstepping decentralized feedforward control scheme, the considered interconnected large-scale nonlinear system in strict feedback form is changed into an equivalent affine large-scale nonlinear system. Subsequently, an optimal decentralized fuzzy adaptive control scheme is constructed. The whole optimal decentralized fuzzy adaptive controller is composed of a decentralized feedforward control and an optimal decentralized control. It is proved that the developed optimal decentralized controller can ensure that all the variables of the control system are uniformly ultimately bounded, and the cost functions are the smallest. Two simulation examples are provided to illustrate the validity of the developed optimal decentralized fuzzy adaptive control scheme.

  3. Decentralized Gauss-Newton method for nonlinear least squares on wide area network

    Science.gov (United States)

    Liu, Lanchao; Ling, Qing; Han, Zhu

    2014-10-01

    This paper presents a decentralized approach of Gauss-Newton (GN) method for nonlinear least squares (NLLS) on wide area network (WAN). In a multi-agent system, a centralized GN for NLLS requires the global GN Hessian matrix available at a central computing unit, which may incur large communication overhead. In the proposed decentralized alternative, each agent only needs local GN Hessian matrix to update iterates with the cooperation of neighbors. The detail formulation of decentralized NLLS on WAN is given, and the iteration at each agent is defined. The convergence property of the decentralized approach is analyzed, and numerical results validate the effectiveness of the proposed algorithm.

  4. Optimization of Heat Exchangers

    International Nuclear Information System (INIS)

    Catton, Ivan

    2010-01-01

    The objective of this research is to develop tools to design and optimize heat exchangers (HE) and compact heat exchangers (CHE) for intermediate loop heat transport systems found in the very high temperature reator (VHTR) and other Generation IV designs by addressing heat transfer surface augmentation and conjugate modeling. To optimize heat exchanger, a fast running model must be created that will allow for multiple designs to be compared quickly. To model a heat exchanger, volume averaging theory, VAT, is used. VAT allows for the conservation of mass, momentum and energy to be solved for point by point in a 3 dimensional computer model of a heat exchanger. The end product of this project is a computer code that can predict an optimal configuration for a heat exchanger given only a few constraints (input fluids, size, cost, etc.). As VAT computer code can be used to model characteristics (pumping power, temperatures, and cost) of heat exchangers more quickly than traditional CFD or experiment, optimization of every geometric parameter simultaneously can be made. Using design of experiment, DOE and genetric algorithms, GE, to optimize the results of the computer code will improve heat exchanger design.

  5. Optimization of Heat Exchangers

    Energy Technology Data Exchange (ETDEWEB)

    Ivan Catton

    2010-10-01

    The objective of this research is to develop tools to design and optimize heat exchangers (HE) and compact heat exchangers (CHE) for intermediate loop heat transport systems found in the very high temperature reator (VHTR) and other Generation IV designs by addressing heat transfer surface augmentation and conjugate modeling. To optimize heat exchanger, a fast running model must be created that will allow for multiple designs to be compared quickly. To model a heat exchanger, volume averaging theory, VAT, is used. VAT allows for the conservation of mass, momentum and energy to be solved for point by point in a 3 dimensional computer model of a heat exchanger. The end product of this project is a computer code that can predict an optimal configuration for a heat exchanger given only a few constraints (input fluids, size, cost, etc.). As VAT computer code can be used to model characteristics )pumping power, temperatures, and cost) of heat exchangers more quickly than traditional CFD or experiment, optimization of every geometric parameter simultaneously can be made. Using design of experiment, DOE and genetric algorithms, GE, to optimize the results of the computer code will improve heat exchanger disign.

  6. Speciation of selenoamino acids, selenonium ions and inorganic selenium by ion exchange HPLC with mass spectrometric detection and its application to yeast and algae

    DEFF Research Database (Denmark)

    Larsen, Erik Huusfeldt; Hansen, M.; Fan, T.

    2001-01-01

    Cation and anion exchange HPLC were used to separate a mixture of 12 selenium species comprising selenoamino acids, selenonium ions and inorganic selenium. The cationic species were separated from each other and from the co-injected anions using a cation exchange column with gradient elution...... by aqueous pyridinium formate at pH similar to 3 as the mobile phase. The anionic species were separated using an anion exchange column with isocratic elution by an aqueous salicylate-TRIS mobile phase at pH 8.5. The separated selenium species were detected as Se-80 by ICP-dynamic reaction cell (DRC...... acid extract of Chlorella algae contained dimethylselenonium propionate (DMSeP), which was verified by HPLC-ES-MS. Se-allylselenocysteine and selenoethionine was detected at the low ng g(-1) concentration level based on co-chromatography with the standard substances spiked to the algal extract....

  7. Algorithms for Lightweight Key Exchange.

    Science.gov (United States)

    Alvarez, Rafael; Caballero-Gil, Cándido; Santonja, Juan; Zamora, Antonio

    2017-06-27

    Public-key cryptography is too slow for general purpose encryption, with most applications limiting its use as much as possible. Some secure protocols, especially those that enable forward secrecy, make a much heavier use of public-key cryptography, increasing the demand for lightweight cryptosystems that can be implemented in low powered or mobile devices. This performance requirements are even more significant in critical infrastructure and emergency scenarios where peer-to-peer networks are deployed for increased availability and resiliency. We benchmark several public-key key-exchange algorithms, determining those that are better for the requirements of critical infrastructure and emergency applications and propose a security framework based on these algorithms and study its application to decentralized node or sensor networks.

  8. Decentralized Electric Vehicle Charging Strategies for Reduced Load Variation and Guaranteed Charge Completion in Regional Distribution Grids

    Directory of Open Access Journals (Sweden)

    Weige Zhang

    2017-01-01

    Full Text Available A novel, fully decentralized strategy to coordinate charge operation of electric vehicles is proposed in this paper. Based on stochastic switching control of on-board chargers, this strategy ensures high-efficiency charging, reduces load variations to the grid during charging periods, achieves charge completion with high probability, and accomplishes approximate “valley-filling”. Further improvements on the core strategy, including individualized power management, adaptive strategies, and battery support systems, are introduced to further reduce power fluctuation variances and to guarantee charge completion. Stochastic analysis is performed to establish the main properties of the strategies and to quantitatively show the performance improvements. Compared with the existing decentralized charging strategies, the strategies proposed in this paper can be implemented without any information exchange between grid operators and electric vehicles (EVs, resulting in a communications cost reduction. Additionally, it is shown that by using stochastic charging rules, a grid-supporting battery system with a very small energy capacity can achieve substantial reduction of EV load fluctuations with high confidence. An extensive set of simulations and case studies with real-world data are used to demonstrate the benefits of the proposed strategies.

  9. Heat exchanger

    International Nuclear Information System (INIS)

    Dostatni, A.W.; Dostatni, Michel.

    1976-01-01

    In the main patent, a description was given of a heat exchanger with an exchange surface in preformed sheet metal designed for the high pressure and temperature service particularly encountered in nuclear pressurized water reactors and which is characterised by the fact that it is composed of at least one exchanger bundle sealed in a containment, the said bundle or bundles being composed of numerous juxtaposed individual compartments whose exchange faces are built of preformed sheet metal. The present addendun certificate concerns shapes of bundles and their positioning methods in the exchanger containment enabling its compactness to be increased [fr

  10. Improving patient activity schedules by multi-agent Pareto appointment exchanging

    NARCIS (Netherlands)

    I.B. Vermeulen (Ivan); S.M. Bohte (Sander); D.J.A. Somefun (Koye); J.A. La Poutré (Han)

    2006-01-01

    textabstractWe present a dynamic and distributed approach to the hospital patient scheduling problem: the multi-agent Pareto-improvement appointment exchanging algorithm, MPAEX. It respects the decentralization of scheduling authorities and is capable of continuously adjusting the different patient

  11. THE DECENTRALIZATION PROCESS IN ROMANIA HAS BEEN AFFECTED BY THE FINANCIAL CRISIS OR NOT; ARGUMENTS IN FAVOR OR AGAINST DECENTRALIZATION IN THE MANAGEMENT OF THE FINANCIAL CRISIS

    Directory of Open Access Journals (Sweden)

    OLIVIA MANOLE

    2012-12-01

    Full Text Available Typically, the decentralization process is extremely complicated and involves many challenges, if we were to take into account local conflicts, the interests of the central government and the complexity of simultaneous decentralization in administrative, political and economic plan. The financial crisis has added another dimension to the complexity of this phenomenon, misbalancing the economy and creating a fiscal pressure both at central and local level. In this context it rises the problem whether the management of the financial crisis can be better realised within a decentralized system or whether it may lay pressure on the return to the centralized government form.

  12. Conformational preludes to the latency transition in PAI-1 as determined by atomistic computer simulations and hydrogen/deuterium-exchange mass spectrometry

    DEFF Research Database (Denmark)

    Petersen, Michael; Madsen, Jeppe B; Jørgensen, Thomas J D

    2017-01-01

    activator inhibitor 1 (PAI-1). We report the first multi-microsecond atomistic molecular dynamics simulations of PAI-1 and compare the data with experimental hydrogen/deuterium-exchange data (HDXMS). The simulations reveal notable conformational flexibility of helices D, E and F and major fluctuations...

  13. Combined cation-exchange and extraction chromatographic method of pre-concentration and concomitant separation of Cu(II) with high molecular mass liquid cation exchanger after its online detection.

    Science.gov (United States)

    Mandal, B; Roy, U S; Datta, D; Ghosh, N

    2011-08-19

    A selective method has been developed for the extraction chromatographic trace level separation of Cu(II) with Versatic 10 (liquid cation exchanger) coated on silanised silica gel (SSG-V10). Cu(II) has been extracted from 0.1M acetate buffer at the range of pH 4.0-5.5. The effects of foreign ions, pH, flow-rate, stripping agents on extraction and elution have been investigated. Exchange capacity of the prepared exchanger at different temperatures with respect to Cu(II) has been determined. The extraction equilibrium constant (K(ex)) and different standard thermodynamic parameters have also been calculated by temperature variation method. Positive value of ΔH (7.98 kJ mol⁻¹) and ΔS (0.1916 kJ mol⁻¹) and negative value of ΔG (-49.16 kJ mol⁻¹) indicated that the process was endothermic, entropy gaining and spontaneous. Preconcentration factor was optimized at 74.7 ± 0.2 and the desorption constants K(desorption)¹(1.4 × 10⁻²) and K(desorption)²(9.8 × 10⁻²) were determined. The effect of pH on R(f) values in ion exchange paper chromatography has been investigated. In order to investigate the sorption isotherm, two equilibrium models, the Freundlich and Langmuir isotherms, were analyzed. Cu(II) has been separated from synthetic binary and multi-component mixtures containing various metal ions associated with it in ores and alloy samples. The method effectively permits sequential separation of Cu(II) from synthetic quaternary mixture containing its congeners Bi(III), Sn(II), Hg(II) and Cu(II), Cd(II), Pb(II) of same analytical group. The method was found effective for the selective detection, removal and recovery of Cu(II) from industrial waste and standard alloy samples following its preconcentration on the column. A plausible mechanism for the extraction of Cu(II) has been suggested. Copyright © 2011 Elsevier B.V. All rights reserved.

  14. Management decentralization and montane forest conditions in Tanzania.

    Science.gov (United States)

    Persha, Lauren; Blomley, Tom

    2009-12-01

    We examined how differences in local forest-management institutions relate to disparate anthropogenic forest disturbance and forest conditions among three neighboring montane forests in Tanzania under centralized, comanaged, or communal management. Institutional differences have been shaped by decentralization reforms. We conducted semistructured interviews with members of forest management committees, local government, and village households and measured anthropogenic disturbance, tree structure, and species composition in forest plots. We assessed differences in governance system components of local institutions, including land tenure, decision-making autonomy by forest users, and official and de facto processes of rule formation, monitoring, and enforcement among the three management strategies. We also assessed differences in frequencies of prohibited logging and subsistence pole cutting, and measures of forest condition. An adjacent research forest served as an ecological reference for comparison of forest conditions. Governance was similar for comanaged and centralized management, whereas communal managers had greater tenure security and decision-making autonomy over the use and management of their forest. There was significantly less illegal logging in the communal forest, but subsistence pole cutting was common across all management strategies. The comanaged forest was most disturbed by recent logging and pole cutting, as were peripheral areas of the larger centralized forest. This manifested in more degraded indicators of forest conditions (lower mean tree size, basal area, density of trees >or= 90 cm dbh, and aboveground biomass and higher overall stem density). Greater tenure security and institutional autonomy of the communal strategy contributed to more effective management, less illegal logging, and maintenance of good forest conditions, but generating livelihood benefits was a challenge for both decentralized strategies. Our results underscore the

  15. Centralization and decentralization in energy systems and associated risks

    International Nuclear Information System (INIS)

    Rehm, W.; Schinner, F.; Kromp, W.

    1998-01-01

    Full text of publication follows: the concept of centralization/decentralization is - considering its widespread use - surprisingly ill-defined. Within the SERF-program (Socio Economic Research on Fusion) an attempt is made to clarify the complex issue of centralization/decentralization and to analyze its expected interaction with electricity production system in the case of nuclear fusion. Fusion research at the time is a multinational, highly centralized undertaking. As a result of this, and due to the expected large energy production units, it is generally expected that fusion will be embedded in a highly centralized, international energy supply system. The degree of centralization of an energy supply system, however, is believed to have for reaching consequences on the stability of the system, and beyond that, on decision making processes in energy policy, as well as democratic structures on the national or even international level. Internationalization might require control instruments that do not exist so far. Amongst other reservations, it is the possibly unwanted consequences of such a highly centralized, international energy supply system that limit acceptance of the fusion technology in the public. Therefore, centralization and decentralization trends of energy supply systems over the years are being looked at, to study their influence on the political and social systems (and vice-versa). In a first step, an attempt is made to classify centralization. Careful analyses and evaluation of available literature revealed a broad spectrum of different forms and degrees of centralization. Various parameters describing energy production and distribution are used. A differentiation of centralization on a technical and on an organizational level seems necessary, although they are not fully independent of each other. Beyond this, the choice of parameters must depend on the questions to be addressed. A new approach defining different levels and degrees of

  16. Decentralized Formation Flying Control in a Multiple-Team Hierarchy

    Science.gov (United States)

    Mueller, Joseph .; Thomas, Stephanie J.

    2005-01-01

    This paper presents the prototype of a system that addresses these objectives-a decentralized guidance and control system that is distributed across spacecraft using a multiple-team framework. The objective is to divide large clusters into teams of manageable size, so that the communication and computational demands driven by N decentralized units are related to the number of satellites in a team rather than the entire cluster. The system is designed to provide a high-level of autonomy, to support clusters with large numbers of satellites, to enable the number of spacecraft in the cluster to change post-launch, and to provide for on-orbit software modification. The distributed guidance and control system will be implemented in an object-oriented style using MANTA (Messaging Architecture for Networking and Threaded Applications). In this architecture, tasks may be remotely added, removed or replaced post-launch to increase mission flexibility and robustness. This built-in adaptability will allow software modifications to be made on-orbit in a robust manner. The prototype system, which is implemented in MATLAB, emulates the object-oriented and message-passing features of the MANTA software. In this paper, the multiple-team organization of the cluster is described, and the modular software architecture is presented. The relative dynamics in eccentric reference orbits is reviewed, and families of periodic, relative trajectories are identified, expressed as sets of static geometric parameters. The guidance law design is presented, and an example reconfiguration scenario is used to illustrate the distributed process of assigning geometric goals to the cluster. Next, a decentralized maneuver planning approach is presented that utilizes linear-programming methods to enact reconfiguration and coarse formation keeping maneuvers. Finally, a method for performing online collision avoidance is discussed, and an example is provided to gauge its performance.

  17. Laboratory-Scale Column Testing Using IONSIV IE-911 for Removing Cesium from Acidic Tank Waste Simulant. 2: Determination of Cesium Exchange Capacity and Effective Mass Transfer Coefficient from a 500-cm3 Column Experiment

    International Nuclear Information System (INIS)

    T.J. Tranter; R.D. Tillotson; T.A. Todd

    2005-01-01

    A semi-scale column test was performed using a commercial form of crystalline silicotitanate (CST) for removing radio-cesium from a surrogate acidic tank solution, which represents liquid waste stored at the Idaho National Engineering and Environmental Laboratory (INEEL). The engineered form of CST ion exchanger, known as IONSIVtmIE-911 (UOP, Mt. Laurel, NJ, USA), was tested in a 500-cm3 column to obtain a cesium breakthrough curve. The cesium exchange capacity of this column matched that obtained from previous testing with a 15-mc3 column. A numerical algorithm using implicit finite difference approximations was developed to solve the governing mass transport equations for the CST columns. An effective mass transfer coefficient was derived from solving these equations for previously reported 15 cm3 tests. The effective mass transfer coefficient was then used to predict the cesium breakthrough curve for the 500-cm3 column and compared to the experimental data reported in this paper. The calculated breakthrough curve showed excellent agreement with the data from the 500-cm3 column even though the interstitial velocity was a factor of two greater. Thus, this approach should provide a reasonable method for scale up to larger columns for treating actual tank waste

  18. Agent-Based Decentralized Control Method for Islanded Microgrids

    DEFF Research Database (Denmark)

    Li, Qiang; Chen, Feixiong; Chen, Minyou

    2016-01-01

    In this paper, an agent-based decentralized control model for islanded microgrids is proposed, which consists of a two-layer control structure. The bottom layer is the electrical distribution microgrid, while the top layer is the communication network composed of agents. An agent is regarded......) a systematic method is presented, which can be used to derive a set of control laws for agents from any given communication network, where only local information is needed. Furthermore, it has been seen that the output power supplied by distributed generators satisfies the load demand in the microgrid, when...

  19. Centralized versus Decentralized Taxation of Mobile Polluting Firms

    OpenAIRE

    Georges A. Tanguay; Nicolas Marceau

    2000-01-01

    We consider a world in which a mobile polluting firm must locate in one of two regions. The regions differ in two dimensions: their marginal cost of pollution and the production cost of the firm. It is shown that under incomplete information on regional marginal costs of pollution, fiscal competition may lead to the sub-optimal location of the firm. We also show that under incomplete information, a sub-optimal location is less likely under centralized than under decentralized taxation. Nous é...

  20. Democratic Decentralization and Empowerment of Local Government Associations in Kerala

    Directory of Open Access Journals (Sweden)

    N Ramakantan

    2009-01-01

    Full Text Available During the past decade the Indian state of Kerala has been successfully carrying out democratic decentralization, and has substantially transformed the functions of local governments in line with the 73rd and 74th Constitutional Amendment Acts, which institutionalised the local government system in India. In particular, formulation and implementation of micro plans with community participation has produced remarkable changes in the dynamics of local development and in the public management of local governments. This initiative for participatory planning at the local level taken by the government of Kerala enormously empowered local communities and the different actors in the local political system.